Tesi sul tema "Mouth diseases – drug therapy"

Magister Pharmaceuticae - MPharm
Neurodegenerative diseases are multifactorial in nature, and taking the complex nature of these disorders into consideration, multi-target directed ligands may present as better options to treat these disorders than the classic ‘one molecule, one target’ approach. Neurotransmitter amines are catabolized by monoamine oxidase A and B (MAO-A and MAO-B), therefore they have been targeted for the treatment of neuropsychiatric and neurodegenerative diseases. Besides offering a potential antidepressant action in PD, MAO-A inhibitors may also provide a symptomatic benefit by reducing MAO-A-catalysed oxidation of dopamine. The oxidative deamination reaction catalyzed by MAO-B is one of the major catabolic pathways of dopamine in the brain. Inhibition of this enzyme leads to enhanced dopaminergic neurotransmission and are currently used in the symptomatic treatment of PD. Furthermore, MAO-B inhibitors may also exert neuroprotective effects by reducing the concentration of potentially hazardous by-products produced by MAO-B-catalysed dopamine oxidation. Apoptosis or programmed cell death occurs in a number of neurodegenerative disorders and it has been proven that inhibition of the executing enzyme, caspases-3, slows down or even stops apoptosis. Having this in mind we focused on the propargylamine function of selegiline and the fluorophenyl function of safinamide, because of their inherent monoamine oxidase inhibitory activities; and isatin as a non-peptide inhibitor of caspase-3. Therefore we attempted to design and synthesize multifunctional hybrid molecules acting simultaneously to halt the apoptotic neuronal breakdown process and eliminate the signs and symptoms of diseases such as PD. Seven novel compounds were successfully synthesized utilizing multistep processes. The synthesis of 5 chlorosulfonyl isatin was accomplished starting from the commercially available isatin in two steps, which were, sulfonation using tetramethylene sulfone and chlorination with POCl₃. Next 5-chlorosulfonyl isatin was conjugated to the fluorophenylamine derivative with the fluoro-function at either the ortho, meta or para position through a nucleophilic substitution reaction on the chlorosulfonyl position. The resultant compounds were coupled on the N position of the isatin function with propargyl bromide, using a microwave synthesis procedure, in a nucleophilic substitution reaction. The structures and purity were confirmed by nuclear magnetic resonance (NMR) and mass spectrometry (MS). In the biological evaluations recombinant human MAO-A, MAO-B and caspase 3 enzymatic assays were performed to determine the activity of the novel compounds at an enzymatic level. The inhibition percentages for these compounds were calculated and plotted against the logarithm 8 of the inhibitor concentrations to obtain a sigmoidal dose-response curve and consequently the IC50 value. The synthesized compounds showed inhibition of the MAO-A, MAO-B and caspase-3 enzymes at low to high micro molar concentrations. The role of the fluorophenylamine moiety in the synthesized compounds was significant for their multifunctional activity as shown by compounds 3 and 4 having good inhibitory activity towards MAO-A, MAO-B and also excellent inhibitory activity against caspase 3, making those ideal candidates for further lead compound development and multifunctional drug design. The introduction of the propargylamine moiety only increased the MAO-A inhibitory activity; this was shown by compounds 7, 8 and 9 which exhibit good MAO-A selectivity with low MAO-B and caspase-3 inhibitory activities.

The use of aminoglycosides in the treatment of the febrile neutropaenic patient with haematological disease is difficult and often suboptimal. This study reviews the available literature to establish therapeutic guidelines in this population and then reports the use of a Bayesian statistics based predictive model to implement and manage therapy in 10 patients. A review of the literature on aminoglycoside Pharmacology and clinical use is essential to determine therapeutic guidelines for this population. Aminoglycosides are amino sugars in glycosidic linkage and are polycations at physiological PH. The antibiotic effect is mediated through inhibition of protein synthesis and disruption of cell membrane integrity. Principal use is in treatment of Gram negative infection although aminoglycosides have activity against some Gram positive organisms including staphylococci. Aminoglycosides are inactive against anaerobes. Acquired resistance is mediated by bacterial enzymatic drug metabolism. Aminoglycosides are nephro- and ototoxic, this is the major constraint in clinical use.

Diabetes mellitus has been associated with both clinical and experimental cardiac dysfunction. Diabetic cardiomyopathy which is characterized by depressed cardiac contractility is accompanied by a variety of biochemical changes in Ca⁺⁺ metabolism. This cardiomyopathy may occur in the presence of normal coronary arteries and normal blood pressure. However, some studies have shown that hypertension is more prevalent among diabetics and can aggravate the cardiovascular abnormalities associated with diabetes. To understand the mechanisms of diabetic cardiomyopathy and consequences of combined hypertension and diabetes, experiments were designed to measure cardiac tissue responses to various inotropic agents in experimental diabetes. Six weeks following streptozotocin (STZ) administration, Wistar, spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats exhibited the 'classical signs' of diabetes which included: hyperglycemia, hypoinsulinemia, hyperlipidemia (except in WKY), and hypothyroidism. Decreased basal atrial rate and increased basal developed force (BDF) suggest a depressed SA node function and an alteration of Ca⁺⁺ utilization by diabetic ventricles. Decreased post quiescent potentiation (PQP) values (except in WKY) in ventricular tissues suggest a diminished amount of releasable Ca⁺⁺ from sarcoplasmic reticulum (SR). Decreased post stimulation potentiation (PSP) values in SHR papillary muscles (PM) are probably suggestive of a depressed sarcolemmal Na⁺-Ca⁺⁺ exchange function in this tissue. Diabetic rats show subsensitivity to β-adrenergic stimulation in ventricular tissues, supersensitivity and hyperresponsiveness to Ca⁺⁺ and α-adrenergic stimulation (except in WKY) in ventricular tissues and left atria (LA) and supersensitivity to BAY K 8644 in SHR LA and hyperresponsiveness to verapamil in ventricular strips. These alterations may be attributed to a change in receptor number and/or a post receptor alteration. Ryanodine decreased the PQP of Wistar and SHR PM and SHR LA in both controls and diabetics. It especially abolished PQP in SHR diabetic tissues, but had no effect on WKY tissues, which may suggest a difference in the SR function in these tissues. SR with impaired Ca⁺⁺ uptake may contribute to these phenomena in diabetic rats. Ryanodine also diminished (PQP + BDF) of SHR LA and (PQP/BDF) of Wistar and SHR PM, ˙but had no effects on control and other diabetic tissues. It appears that ryanodine has some influence on the Na⁺-Ca⁺⁺ exchange generated by sarcolemma (SL) of certain diabetic tissues. Further experiments are required to clarify this. SHR diabetic rats had greater changes in most of the measurements such as hyperlipidemia, depressed PQP and PSP values, and altered drug responses. This model exhibited very high mortality as compared to Wistar and WKY diabetic rats. As has been shown previously, the combination of hypertension and diabetes exerts a synergistic effect on the cardiac dysfunction in this model, and that altered lipid metabolism, SL and SR function are all involved in the development of cardiomyopathy. WKY diabetic rats, on the other hand, exhibited no significant changes in blood lipids, or in response to phenylephrine or to Ca⁺⁺ (LA) stimulation. Lack of change in these factors may explain the relatively normal cardiac function of this model as measured previously.
Pharmaceutical Sciences, Faculty of
Graduate

The aim of this study was to develop drug delivery system based on poly(lactic acid-co-glycolic acid) (PLGA) nanoparticles (NPs) to achieve personalized treatment of selected skin disorders, like photo-aging, psoriasis and atopic dermatitis. Dead Sea Water (DSW) and Retinyl Palmitate (RP) were used as active agents and they were loaded in PLGA NPs prepared either as spheres or capsules by o/w or w/o/w methods. MgCl2 and bovine serum albumin (BSA) served as model active compounds. The diameter of the NPs was found to be in the range of 280 - 550 nm. The entrapment efficiency (E.E.) was less than 1% for RP, DSW and MgCl2, and 41% for BSA. Loading of Cl- together with BSA doubled the E.E. value of Cl- . In situ release studies showed a burst in the first day and more than 85% of the chloride content was released within a week. When the macromolecule BSA was encapsulated, a much slower and triphasic release profile was observed which continued for up to 80 days. In vitro tests were performed using L929 fibroblast cells. Results of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) test revealed that none of the NPs were cytotoxic. Additionally, all particles were hemocompatible with hemolytic activity <
1.5%. L929 fibroblast and Saos 2 human osteosarcoma cells were used to study the uptake of NPs by the cells. Particles accumulate near the nucleus. The characterization and cell viability tests, and drug release behavior indicate the suitability of these NPs for further testing to develop a patient specific skin diseases treatment approach.

Background: Idiopathic pulmonary fibrosis (IPF) is the most common and lethal among diffuse fibrosing interstitial lung diseases (ILD). Beyond lung transplantion, the therapeutic approach relies on antifibrotic treatment: pirfenidone and nintedanib are the only pharmaceutical drugs approved for IPF, since they have demonstrated to significantly reduce disease progression rate. Still, no solid data has been published to compare these two drugs as well as few studies have investigated their potential efficacy on familial pulmonary fibrosis (FPF) and progressive fibrosing ILD (PF-ILD) in a real-life setting. Methods: we collected clinical, functional and radiological data from all patients affected with IPF and PF-ILDs that have been treated with pirfenidone and nintedanib at Referral Centre of Siena from 2011 to 2020. The aim of the research was to compare effectiveness of the two drugs in terms of mortality and disease progression in our population. Results: no significant differences in mortality and progression-free survival were observed between pirfenidone and nintedanib subgroup. Both drugs significantly reduce FVC and DLCO decline rate in respect with pretreatment period. Similar data was observed in the PF-ILD subgroup, while FPF patients showed no significant benefit from antifibrotic treatment in terms of disease progression. Pirfenidone was more effective than nintedanib in preserving FVC in FPF subgroup. Conclusions: our research study, conducted in a large cohort through a almost decennial time of observation, confirmed the reliable and substantially similar efficacy of pirfenidone and nintedanib in improving life expectancy and progression-free survival of IPF patients. FPF appeared to be less responsive to antifibrotics, but pirfenidone showed a better performance than nintedanib on this field. PF-ILD patients showed a analogue clinical course of IPF subjects in our study: the effectiveness of pirfenidone and nintedanib was reliable and similar, supporting their future use in clinical practice.

Thesis (M.Tech.)(Nuclear Medicine) -- Central University of Technology, free State, 2007
In the South African Health Services (SAHS) it is each health worker’s responsibility to find ways to reduce health care cost and improve health service to the public. The measurement of radioactive iodine uptake (RAIU) by the thyroid gland for diagnostic purposes has been used as early as the 1940s. The 24-hour (hr) iodine-131 (131I) uptake measurement is traditionally used for the calculation of the 131I administered activity for therapy dosage. This entails that the patient’s hospitalisation is prolonged, which increases the costs. The literature also indicates that the 24-hr 131I uptake value can be discarded and only the 6-hr 131I uptake measurement is needed to calculate administered activity for therapeutic dosages for Graves’ patients. Therefore, if it can be confirmed that the 6-hr 131I uptake measurement alone is needed, the SAHS could decrease hospitalisation costs. The overall goal of the investigation was to analyse the 6-hr and 24-hr 131I uptake measurements of patients with Graves’ disease at the Universitas Hospital. The aim was to determine the relationship between the 6-hr and 24- hr RAIU values to establish the therapeutic dosage for Graves’ disease. To achieve the aim, three objectives were set. First, to serve as a background to the investigation, a literature survey relating to the RAIU measurements of patients with Graves’ disease was made. Second, a retrospective analysis was performed by collecting the 6-hr and 24-hr 131I uptake measurements of patients with proven Graves’ disease at the Universitas Nuclear Medicine Department (UNMD). Finally, the data obtained from the retrospective analysis was analysed, summarised and compared to answer the investigation questions. The investigation group included patients with confirmed Graves’ disease who had undergone both the 6- and 24-hr 131I RAIU at the Universitas Hospital from the beginning of 2004 to the end of 2005. Graves’ disease is confirmed by the following factors at the UNMD, namely: Suppressed TSH, elevated T4 and T3 values, an increased uptake on the 99mTc-pertechnetate scan and increased 6- and 24-hr 131I RAIU values. The UNMD statistics show that 178 patients were diagnosed with Graves’ disease during this period. The patients of the investigation group included both male and female patients from different races, ranging from 15-75 years. In order to increase the validity of the investigation, all factors that could influence the accuracy of the 131I thyroid uptake test were excluded. After the exclusion and inclusion criteria had been applied, the final investigation group was made up of 124 Graves’ disease patients. The data obtained from the patient files was noted on the different data sheets (see Appendix A) for further analysis. The information from these data sheets was then used to obtain the investigation results. The Department of Biostatistics of the University of the Free State (UFS) was consulted for recommendations regarding the management of data and the processing of results. All values were summarised by means and Standard Deviations (SD) or percentiles. Mean or median differences were calculated with a 95% Confidence Interval (CI). A regression analysis was made between the 6-hr and 24-hr 131I RAIU values. The highest RAIU value is the best to calculate the therapeutic dosage, as this gives a true reflection of the thyroid function of a Graves’ disease patient. In the investigation group the median of the 24-hr 131I RAIU values was higher than the 6-hr 131I RAIU values. The findings showed that the 24-hr 131I RAIU in most of the investigation group was the highest value and most effective to calculate the 131I therapeutic dosage. At a time when research-based practice is taking on an increasingly important role, it is essential for nuclear medicine departments to make evidence-based recommendations. This investigation found that the correlation between the 6-hr and 24-hr RAIU clearly justified the cost spent on Graves’ disease patients who must stay overnight for the 24-hr 131I RAIU procedure.

Viral proteases have been shown to be effective targets of anti-viral therapies for human immunodeficiency virus (HIV) and hepatitis C virus (HCV). However, under the pressure of therapy including protease inhibitors, the virus evolves to select drug resistance mutations both in the protease and substrates. In my thesis study, I aimed to understand the mechanisms of how this protease−substrate co-evolution contributes to drug resistance. Currently, there are no approved drugs against dengue virus (DENV); I investigated substrate recognition by DENV protease and designed cyclic peptides as inhibitors targeting the prime site of dengue protease. First, I used X-ray crystallography and subsequent structural analysis to investigate the molecular basis of HIV-1 protease and p1-p6 substrate coevolution. I found that co-evolved p1-p6 substrates rescue the HIV-1 I50V protease’s binding activity by forming more van der Waals contacts and hydrogen bonds, and that co-evolution restores the dynamics at the active site for all three mutant substrates. Next, I used aprotinin as a platform to investigate DENV protease–substrate recognizing pattern, which revealed that the prime side residues significantly modulate substrate affinity to protease and the optimal interactions at each residue position. Based on these results, I designed cyclic peptide inhibitors that target the prime site pocket of DENV protease. Through optimizing the length and sequence, the best inhibitor achieved a 2.9 micromolar Ki value against DENV3 protease. Since dengue protease does not share substrate sequence with human serine proteases, these cyclic peptides can be used as scaffolds for inhibitor design with higher specificity.

Viral proteases have been shown to be effective targets of anti-viral therapies for human immunodeficiency virus (HIV) and hepatitis C virus (HCV). However, under the pressure of therapy including protease inhibitors, the virus evolves to select drug resistance mutations both in the protease and substrates. In my thesis study, I aimed to understand the mechanisms of how this protease−substrate co-evolution contributes to drug resistance. Currently, there are no approved drugs against dengue virus (DENV); I investigated substrate recognition by DENV protease and designed cyclic peptides as inhibitors targeting the prime site of dengue protease. First, I used X-ray crystallography and subsequent structural analysis to investigate the molecular basis of HIV-1 protease and p1-p6 substrate coevolution. I found that co-evolved p1-p6 substrates rescue the HIV-1 I50V protease’s binding activity by forming more van der Waals contacts and hydrogen bonds, and that co-evolution restores the dynamics at the active site for all three mutant substrates. Next, I used aprotinin as a platform to investigate DENV protease–substrate recognizing pattern, which revealed that the prime side residues significantly modulate substrate affinity to protease and the optimal interactions at each residue position. Based on these results, I designed cyclic peptide inhibitors that target the prime site pocket of DENV protease. Through optimizing the length and sequence, the best inhibitor achieved a 2.9 micromolar Ki value against DENV3 protease. Since dengue protease does not share substrate sequence with human serine proteases, these cyclic peptides can be used as scaffolds for inhibitor design with higher specificity.

The Hepatitis C Virus (HCV) is a global health problem as it afflicts an estimated 170 million people worldwide and is the major cause of viral hepatitis, cirrhosis and liver cancer. HCV is a rapidly evolving virus, with 6 major genotypes and multiple subtypes. Over the past 20 years, HCV therapeutic efforts have focused on identifying the best-in-class direct acting antiviral (DAA) targeting crucial components of the viral lifecycle, The NS3/4A protease is responsible for processing the viral polyprotein, a crucial step in viral maturation, and for cleaving host factors involved in activating immunity. Thus targeting the NS3/4A constitutes a dual strategy of restoring the immune response and halting viral maturation. This high priority target has 4 FDA approved inhibitors as well as several others in clinical development. Unfortunately, the heterogeneity of the virus causes seriously therapeutic challenges, particularly the NS3/4A protease inhibitors (PIs), which suffer from both the rapid emergence of drug resistant mutants as well as a lack of pan-genotypic activity. My thesis research focused on filling two critical gaps in our structural understanding of inhibitor binding modes. The first gap in knowledge is the molecular basis by which macrocyclization of PIs improves antiviral activity. Macrocycles are hydrophobic chains used to link neighboring chemical moieties within an inhibitor and create a structurally pre-organized ligand. In HCV PIs, macrocycle come in two forms: a P1 - P3 and P2 - P4 strategy. I investigated the structural and thermodynamic basis of the role of macrocyclization in reducing resistance susceptibility. For a rigorous comparison, we designed and synthesized both a P1 - P3 and a linear analog of grazoprevir, a P2 - P4 inhibitor. I found that, while the P2 - P4 strategy is more favorable for achieving potency, it does not allow the inhibitor sufficient flexibility to accommodate resistance mutations. On the other hand, the P1 - P3 strategy strikes a better balance between potency and resistance barrier. The second gap my thesis addresses is elucidating the structural basis by which highly potent protease inhibitors function in genotype 1 but not in genotype 3, despite having an 87% sequence similarity. After mapping the amino acids responsible for this differential efficacy in genotypes 1 and 3, I engineered a 1a3a chimeric protease for crystallographic studies. My structural characterization of three PIs in complex with both the 1a3a and genotype 1 protease revealed that the loss of inhibitor efficacy in the 1a3a and GT-3 proteases is a consequence of disrupted electrostatic interactions between amino acids 168 and 155, which is critical for potent binding of quinoline and isoindoline based PIs. Here, I have revealed details of molecular and structural basis for the lack of PI efficacy against GT-3, which are needed for design of pan-genotypic inhibitors.

Antiviral drug resistance is a major problem in the treatment of viral infections, including influenza and hepatitis C virus (HCV). Influenza neuraminidase (NA) is a viral sialidase on the surface of the influenza virion and a primary antiviral target in influenza. Two subtypes of NA predominate in humans, N1 and N2, but different patterns of drug resistance have emerged in each subtype. To provide a framework for understanding the structural basis of subtype specific drug resistance mutations in NA, we used molecular dynamics simulations to define dynamic substrate envelopes for NA to determine how different patterns of drug resistance have emerged in N1 and N2 NA. Furthermore, we used the substrate envelope to analyze HCV NS3/4A protease inhibitors in clinical development. In addition, influenza hemagglutinin (HA) is a primary target of neutralizing antibodies against influenza. Novel broadly neutralizing antibodies (BnAbs) against the stem region of HA have been described and inhibit several influenza viral subtypes, but antibody neutralization escape mutations have emerged. We identified potential escape mutations in broadly neutralizing antibody F10 that may impact protein dynamics in HA that are critical for function. We also solved crystal structures of antibody fragments that are important for understanding the structural basis of antibody binding for influenza BnAbs. These studies can inform the design of improved therapeutic strategies against viruses by incorporating an understanding of structural elements that are critical for function, such as substrate processing and protein dynamics, into the development of novel therapeutics that are robust against resistance.

Multiple modulating genes and environmental factors have been implicated in the pathogenesis of high-altitude pulmonary edema (HAPE). However, at the present time, there exists an incomplete understanding of the molecular mechanisms and pathways which underlie constitutional susceptibility. Genome-wide measurements of gene expression in peripheral blood mononuclear cells (PBMCs) were performed using microarray technology. Comparison of gene expression profiles of HAPE-susceptible and resistant individuals resulted in the identification of several previously undescribed candidate genes. RhoA and Rho-kinase (ROCK), regulators of vascular smooth muscle contraction, were differentially regulated in the HAPE-susceptible cohort, as compared to both HAPE-resistant patients with acute mountain sickness (AMS+) and healthy controls (p=0.0014; p=0.0020). Furthermore, biological pathways involving RhoA and Rho-kinase were strongly upregulated in subjects with HAPE. These findings represent the first description of the RhoA/Rho-kinase signaling pathway in HAPE. Currently, few pharmacologic therapies have been demonstrated to be effective in the prevention and treatment of HAPE. The results of this study provide early evidence that Fasudil, a selective Rho-kinase inhibitor, may represent a novel therapeutic intervention effective in the prevention and/or treatment of high-altitude pulmonary edema.

The description and interpretation of Latinas’ experience with chemotherapyinduced premature menopause from breast cancer treatment were explored in this study, which utilized an interpretive descriptive method from a feminist lens, and Knobf’s (1998, 2002) “Carrying on” theory. The specific aims of the study and the interview questions were guided by the state of the science literature. Overall, the impact of physiological effects, psychosocial effects, barriers, influencing factors that made their experience easier or harder, and how participants adjusted to a cancer diagnosis, treatment course, and menopause transition were described as bigger than the menopause experience alone. Participants also described a period of uncertainty or “ever-changing landscape” that began at the time of diagnosis and continued through survivorship. The impact of information, access to healthcare, acculturation levels, support, and a sense of control were elucidated as important factors in “working through” the experience. A range of collateral data sources were employed. Study limitations and future implications for practice, research, and health policy were demarcated.

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas
As Doenças Negligenciadas ou doenças tropicais, advêm de um conjunto de factores biológicos, ecológicos e evolutivos, com uma elevada incidência nos países de clima tropical. A situação económica desfavorável e o baixo desenvolvimento, estão também directamente relacionados com a sua ocorrência. Estas doenças representam um encargo significativo para a saúde em certas partes do mundo. Os medicamentos disponíveis no tratamento das mesmas não são capazes de atender às suas necessidades clínicas. Como tal, tem havido um esforço significativo para o desenvolvimento de novos sistemas terapêuticos ao longo dos últimos anos. Estes desenvolvimentos têm sido liderados por várias instituições ou agências governamentais com o intuito de obter progressos a concepção de novos fármacos contra estas afecções. Os temas abordados neste estudo incidirão essencialmente sobre a Leishmaniose e a Doença de Chagas. Ambas são causadas por diferentes protozoários, desenvolvendo diferentes manifestações e sintomatologia, dependendo da espécie. Os sistemas terapêuticos (ST) convencionais apresentam inúmeras resistências, pelo que este estudo visa a descrição de novos sistemas terapêuticos (NST) com o intuito de melhorar a biodisponibilidade dos fármacos no respectivo local de acção. Entre eles distinguem-se os lipossomas, niossomas, nanopartículas de lípidos sólidos, nanopartículas poliméricas, nanoemulsões, micelas, entre outros que serão abordados ao longo deste trabalho. Neglected diseases or tropical disorders, come from a number of biological, ecological and evolutionary factors, with high incidence in tropical countries. The unfavorable economic situation and the low development, are also directly related to its occurrence. These diseases represent a significant burden to health in certain parts of the world. The drugs available for treating these diseases are not able to satisfy their clinical needs. However, over the last years, there has been a significant effort at developing DDS. Several institutions or governmental agencies have been leading these developments in order to obtain progress when developing new drugs against these diseases. The discussed themes in this study will coincide essentially on leishmaniasis and Chagas disease. Both are caused by different protozoa, and develop different manifestations and symptoms, according to the specific species. The conventional therapeutic systems already present a lot of resistances, and the intent of this study is the discovery of DDS looking to improve the drugs bioavailability in the respective place of action. These include liposomes, niosomes, solid lipid nanoparticles, polymeric nanoparticles, nanoemulsions, micelles, among others that will be addressed throughout this work.

Dor neuropática e neuropatia causadas pelo quimioterápico oxaliplatina são comuns e causam restrições às atividades funcionais e qualidade de vida. Muitos estudos têm quantificado e qualificado esses sintomas, porém raramente utilizando amostra expressiva e instrumentos validados e específicos para este fim. Neste estudo é proposta uma análise ampla, por instrumentos que quantificam e qualificam a dor e neuropatia relacionadas à oxaliplatina e suas características associadas. Objetivos: Identificar a prevalência de dor neuropática e neuropatia periférica em doentes com câncer colorretal recebendo tratamento com oxaliplatina durante os seis meses de tratamento quimioterápico e após seguimento (de 3 a 6 meses); avaliar, comparar e descrever as características da dor e neuropatia nesta população e avaliar seu impacto nas atividades de vida diária e qualidade de vida. Métodos: Foram incluídos 110 doentes (média 55,6 anos) com câncer colorretal durante o tratamento quimioterápico com oxaliplatina e seguidos por 3 a 6 meses após quimioterapia. Os doentes responderam ao questionário sócio-demográfico e a questionários específicos para dor e neuropatia antes da quimioterapia e bimestralmente por até seis meses durante a quimioterapia e em avaliação de seguimento realizada de 3 a 6 meses após o término da quimioterapia. Os instrumentos utilizados foram: Questionário de Qualidade de Vida da Organização Européia para Pesquisa e Tratamento do Câncer - C30 (EORTC QLQ-C30); Questionário de Dor McGill reduzido (QDMR), Inventário de Sintomas de Dor Neuropática (ISDN), Inventário Breve de Dor (BPI-Brief Pain Inventory) Questionário de Dor Neuropática 4 (DN4), Escala Hospitalar de Ansiedade e Depressão (HADS), Escore Total de Neuropatia (TNS - Total Neuropathy Score). Resultados: Em geral, a dor e neuropatia aumentaram durante o período de quimioterapia e diminuíram após fim do tratamento, permanecendo em níveis significativamente mais elevados após o fim do tratamento quimioterápico. A média de intensidade de dor (dor mais intensa) segundo o IBD foi 2,54 na V3 (após 4 meses de tratamento com oxaliplatina). A dor foi do tipo neuropática em 21,67% dos doentes ao fim da quimioterapia e em 10,00% após fim do seguimento. As médias segundo o ISDN foram 0,67 no basal, 18,67 na V2, 17,77 na V3, 17,44 após quimioterapia e 11,03 após seguimento. A característica da dor mais frequente foi em choque elétrico, enjoada e incômoda segundo o QDMR e segundo o ISDN foram choque elétrico, frio doloroso e dormência. A qualidade de vida sofreu impacto negativo. Conclusões: Dor neuropática foi prevalente após a quimioterapia e após seguimento e se associou com interferência negativa sobre as atividades diárias. Estes dados poderão auxiliar o desenvolvimento de tratamentos individualizados da neuropatia relacionada à oxaliplatina
Neuropathic pain and neuropathy caused by oxaliplatin chemotherapy are common and cause restrictions in activities of daily living and quality of life. Many studies have quantified and qualified these symptoms but only rarely used a comprehensive sample of patients and employed validated and specific instruments for pain assessment. In this study we proposed a comprehensive analysis by instruments that quantify and qualify the pain and neuropathy and its characteristics. Aim of Investigation: To identify the prevalence of neuropathic pain and peripheral neuropathy in patients with colorectal cancer receiving treatment with oxaliplatin during the six months of chemotherapeutic treatment and after follow-up (3 to 6 months); to evaluate, compare and describe pain and peripheral neuropathy characteristics in this population and evaluate their impact on activities of daily living. Methods:110 patients (mean 55.6 years) with colorectal cancer were included during the six months of chemotherapy with oxaliplatin and follow-up (3 to 6 months) after chemotherapeutic treatment. Patients answered socio-demographic questionnaires and specific assessment tools for pain and neuropathy evaluation at the baseline visit and every 2 months during chemotherapy and after follow-up (3-6 months). The instruments used were: The European Organization for Research and Treatment of Cancer QLQ-C30 (EORTC QLQ-C30); Reduced McGill Pain Questionnaire (MPQ), Neuropathic Pain Symptom Inventory (NPSI), Brief Pain Inventory (BPI) Neuropathic Pain Diagnostic Questionnaire (DN4), Hospital Anxiety and Depression Scale (HADS), Total Neuropathy Score (TNS), Results: In general, pain and neuropathy intensity increased during chemotherapy and decreased after the end of treatment (follow-up). The most severe pain according to the BPI was 2.54 in V3 (after 4 months treatment with oxaliplatin). Pain was neuropathic in 21.67% right after chemotherapy and in 10.00% after follow-up according to the DN4. The average sum of neuropathic pain symptoms according to the NPSI were 0.67 in V1, 18.67 in V2, 17.77 in V3, 17.44 after chemotherapy and 11.03 after follow-up. The most common characteristics of the pain was electric shocks, nauseating and fearful, according to MPQ and according to NPSI were electric shock, evoked by cold stimuli and tingling. Conclusions: Patients treated with oxaliplatin had significant pain and neuropathy that negatively interfeared with daily activities. These data may help tailor individualized treatment of chemotherapy related neuropathic pain

INTRODUÇÃO: A neuropatia periférica induzida por quimioterapia (NPIQ) está entre os efeitos colaterais mais comuns decorrentes da quimioterapia antineoplásica e uma das principais causas da redução da dose ou interrupção do tratamento. Os sintomas mais prevalentes são a dor e a parestesia, acarretando desconfortos crônicos e perda de habilidades funcionais, interferindo negativamente na qualidade de vida dos pacientes. Estudos recentes têm avaliado o uso da Estimulação Elétrica Nervosa Transcutânea (TENS) nesta patologia, apresentando evidências positivas na redução da dor, porém seu efeito nos sintomas de parestesia e nas atividades de vida diária destes pacientes ainda não foram avaliados. OBJETIVO: Investigar os efeitos da Estimulação Elétrica Nervosa Transcutânea (TENS) nos sintomas de dor, parestesia e nas atividades de vida diária da NPIQ em indivíduos com diagnóstico de câncer de mama e colorretal, submetidos ao tratamento de quimioterapia. MÉTODOS: Trata-se de um ensaio clínico duplo-cego, controlado, randomizado e multicêntrico, com abordagem quantitativa, em pacientes submetidos ao tratamento de quimioterapia, contendo em seu protocolo os seguintes quimioterápicos: paclitaxel e oxaliplatina. Os sujeitos da pesquisa utilizaram o dispositivo terapêutico TENS com modulação de frequência entre 7 e 75 Hz na região distal dos membros, no local de maior desconforto, com intervenções diárias de 60 minutos, durante três ciclos de quimioterapia (45 dias). Os participantes foram divididos em dois grupos: grupo TENS ativa (GTA) e grupo TENS placebo (GTP). A avaliação dos efeitos da TENS foi medida através dos seguintes instrumentos: a Escala Visual Analógica (EVA) para avaliar os sintomas de dor e parestesia e Questionário de Neurotoxicidade Induzida por Anti-neoplásicos (QNIA) para avaliação dos sintomas da NPIQ. RESULTADOS: Finalizaram a pesquisa 24 pacientes. Não se observou uma diferença significativa entre os 2 grupos no que se refere ao desfecho primário de redução dos sintomas de dor (p = 0.666), parestesia (p = 0,673) e impacto da TENS na frequência dos sintomas (p = 0,5906) e atividades de vida diária (p = 0,8565). CONCLUSÃO: Estes resultados sugerem que a TENS aplicada no modo de modulação de frequência não foi eficaz para melhorar os sintomas de neuropatia periférica induzida por quimioterapia, durante os ciclos de quimioterapia. Não houve, porém, agravamento dos sintomas em ciclos subsequentes ao início dos sintomas da doença
BACKGROUND: Peripheral neuropathy induced by chemotherapy (PNIC) is amongst the most common side effects derived from antineoplastic chemotherapy and one of the principal causes of dose reduction or treatment interruption. The most prevalent symptoms are pain and numbness, resulting from chronic discomfort to loss of functional abilities, negatively affecting quality of life and autonomy of patients. Recent studies have evaluated the use of Transcutaneous Electrical Nerve Stimulation (TENS) in this disease, pointing to evidence of pain reduction, but its effect on symptoms of paresthesia and in daily life activities have not yet been evaluated. OBJECTIVE: To investigate the effects of Transcutaneous Electrical Nerve Stimulation (TENS) for reducing the symptoms of pain, paresthesia and the daily activities of PNIC in patients diagnosed with breast cancer and colorectal cancer undergoing chemotherapy treatment. METHODS: It is a double-blind, controlled, randomized, multicenter clinical trial with a quantitative approach in a sample of 24 patients undergoing chemotherapy treatment, containing in its protocol the following chemotherapeutic agents: paclitaxel and oxaliplatin. The research subjects used the TENS therapeutic device with frequency modulation between 7 and 75 Hz in the distal limb, on the location of greatest discomfort with daily interventions lasting 60 minutes for three chemotherapy cycles (45 days). Participants were divided into two groups: active TENS group (ATG) and placebo TENS group (PTG). The assessment of the effects of TENS was measured by the following instruments: The Visual Analogue Scale (VAS) to assess the symptoms of pain and numbness and Questionnaire for Neurotoxicity Induced by Anti-neoplastic (QNIA) to assess the symptoms of PNIC. RESULTS: A 24-patient study was completed. There was no significant difference between the two groups regarding the primary endpoint of reduced pain symptoms (p = 0.666) and paresthesia (p = 0.673), neither any measurable impact of TENS in the frequency of symptoms (p = 0.5906) or activities of daily living (p = 0.8565). CONCLUSION: These results suggest that TENS applied in frequency modulation mode is not effective for ameliorating the symptoms of peripheral neuropathy induced by chemotherapy during chemotherapy cycles. There was, however, no worsening of symptoms in subsequent cycles after the onset of symptoms

Background & Aims: Despite advances in surgical care and chemotherapeutic regimens, the five-year survival rate for Stage IV Hepatoblastoma (HB), the predominant pediatric liver tumor, remains at 27%. YAP1 and β-Catenin co-activation occurs in 80% of children’s HB; however, a lack of conditional genetic models precludes exploration of tumor maintenance and therapeutic targets. Thus, the clinical need for a targeted therapy remains unmet. Given the predominance of YAP1 and β-catenin activation in children’s tumors, I sought to evaluate YAP1 as a therapeutic target in HB. Approach & Results: Herein, I engineered the first conditional murine model of HB using hydrodynamic injection to deliver transposon plasmids encoding inducible YAP1S127A, constitutive β-CateninDelN90, and a luciferase reporter to murine liver. Tumor regression was evaluated using in vivo bioluminescent imaging, and tumor landscape characterized using RNA sequencing, ATAC sequencing and DNA foot-printing. Here I show that YAP1 withdrawal in mice mediates >90% tumor regression with survival for 230+ days. Mechanistically, YAP1 withdrawal promotes apoptosis in a subset of tumor cells and in remaining cells induces a cell fate switch driving therapeutic differentiation of HB tumors into Ki-67 negative “hbHep cells.” hbHep cells have hepatocyte-like morphology and partially restored mature hepatocyte gene expression. YAP1 withdrawal drives formation of hbHeps by modulating liver differentiation transcription factor (TF) occupancy. Indeed, tumor-derived hbHeps, consistent with their reprogrammed transcriptional landscape, regain partial hepatocyte function and can rescue liver damage in mice. Conclusions: YAP1 withdrawal, without modulation of oncogenic β-Catenin, significantly regresses hepatoblastoma, providing the first in vivo data to support YAP1 as a therapeutic target for HB. Modulating YAP1 expression alone is sufficient to drive long-term regression in hepatoblastoma because it promotes cell death in a subset of tumor cells and modulates transcription factor occupancy to reverse the fate of residual tumor cells to mimic functional hepatocytes.

A dissertation submitted to the Faculty of Health Science, University of the Witwatersrand, Johannesburg, in fulfillment of the requirements for the degree of Master of Science in medicine. Johannesburg 2014
Introduction: The presence of pain during sleep causes patients with chronic daily pain, such as in Rheumatoid and Osteoarthritis, to experience insomnia, fragmented sleep and an increased number of night-time awakenings. This poor sleep results in an increased sensitivity to pain during the day. The effect of improving sleep on pain, sleep and mood after taking Zolpidem MR was the aim of this study. Methods: 11 patients from Chris Hani Baragwanath Hospital in Soweto South Africa who reported insomnia and daily pain spent 4 weeks in this crossover design, double blinded, placebo controlled study. Week 1- baseline, week 2 and 4 were Intervention weeks – either placebo or Zolpidem MR, week 3 was a Washout week. Data collected included actigraphy, McGill Pain Questionnaire, PSQI, BDI, physical activity questionnaire and daily sleep and pain diaries containing VAS scales for sleep and pain. Results: No significant changes were found in the pain or physical activity levels in any of the patients. Sleep quality, as measured by an isolated PSQI question, was improved by Zolpidem MR (p=0.0075). PSQI was decreased in the final week of the study compared to baseline (8.7 vs. 11.3, p=0.0106) and BDI was lower in week 4 than baseline (7.7 vs. 15.85, p=0.0003), BDI was also lower in week 4 compared to week 2 (7.7 vs. 12.8, p<0.05). However the changes in PSQI and BDI were a result of the order of the weeks, with patients interacting with the researcher and were not due to either Zolpidem MR or placebo. Anecdotal reports include feeling more energised and capable of living life. Conclusion: This study has shown that human interaction is an important component of treatment for insomnia and chronic pain as there is a positive effect on sleep disruption and depression.

A thesis submitted to the Faculty of Health Sciences, University of the Witwatersrand, in fulfilment of the requirements for the degree of Doctor of Philosophy Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, South Africa Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, South Africa Johannesburg 2017.
Psoriasis vulgaris is a chronic, hyper-proliferative skin condition which affects the patient’s quality of life. The treatment strategy involves long term use of drugs that maintain the condition, however; playing a pivotal negative role in patient compliance. A constructive development in the design of treatment addressing the disease should focus on the challenges faced by current designs. Hence, cellular internalization and trans-barrier transport of nanoparticles can be manipulated on the basis of the physicochemical and mechanical characteristics of nanoparticles to enhance the treatment options of the condition by reducing dosing and increasing the healing due to intracellular drug delivery. Dictating these characteristics allows for the control of the rate and extent of cellular uptake, as well as delivering the drug-loaded nanosystem intra-cellularly which is imperative for drugs that require a specific cellular level to exert their effects, as is with psoriasis. Additionally, physicochemical characteristics of the nanoparticles should be optimal for the nanosystem to bypass the natural restricting phenomena of the body and act therapeutically at the targeted site. Neo-geometric copper nanoparticles (CuNPs) in the biomedical application ascertained skin permeation and retention of the CuNPs as a drug delivery system. The approach to the use of the nanocrystal exploited the shape properties as a function of enhanced cellular uptake and the copper in the inflamed psoriatic environment acted as a cytotoxic agent against hyper-proliferating keratinocytes. A Self-Oscillating Polymeric Network (SOPN) served as a vehicle for the topical delivery of the geometric CuNPs in addition to its oscillating phenomenon to promote the permeation of the active nanoparticles across the rate limiting barrier of the skin, the stratum corneum. This twofold system adequately targets the key limitations in addressing psoriasis. A statistical experimental design comprising a full factorial model for the optimization of the geometric CuNPs and Box-Behnken design applied on the SOPN served as a refining factor to achieve stable, homogenous, geometric nanoparticles using a one-pot method for the systematic optimization of the geometric CuNPs. The optimization of the SOPN involved amplitude and duration of the oscillations, permeation kinetics and cytotoxicity. After optimization of the nano-shapes and oscillations of the SOPN, extensive ex vivo cellular internalization studies were conducted to elucidate the effect of geometric CuNPs on uptake rates; in addition to the vital toxicity assays to further understand the cellular effect of geometric CuNPs as a drug delivery system. Complementing the geometry analysis; volume, surface area, orientation to the cell membrane and colloidal stability were also addressed. The SOPN was also investigated ex vivo for its biocompatibility to determine the LD50 and permeation kinetics. The in vivo study probed the nanosystem embedded in the innovative SOPN to stimulate the permeation of the CuNPs across the stratum corneum of the induced psoriasiform-plaque in a BALB/c mouse model. The results confirmed an optimized CuNPs-loaded SOPN topical system with promising plaque thickness reduction when compared with a commercial gold standard in the treatment of the skin condition. This novel system can be safely used with less frequent, lower dosing and no odour, therefore promoting patient compliance.
MT2017

Thesis (M. Pharm.)--University of the Witwatersrand, Faculty of Health Sciences, 2013.
Ocular diseases of the anterior segment are ubiquitous, especially among elderly patients. The development of novel drug delivery systems on the journey for improved treatment is therefore imperative. Aside from anatomical and physiological barriers of the eye, the actual dosage form plays a crucial role. Although liquid eye drops are the first-choice dosage form, the shortcomings do not go unnoticed. In an attempt to circumvent these drawbacks, a novel instantly soluble eye drop device was developed. The system aimed to provide an easier administration form, comfort for the patient and improve drug bioavailability to anterior chamber. This was a steer toward attaining patient-convenience and compliance which are critically challenging factors. Preformulatory studies allowed for the screening and selection of candidate components and key processing conditions. Hydrophilic polymers and excipients were selected for attainment of small, rapid disintegrating yet robust matrices via lyophilization of solutions. Design of experiments generated formulations by means of a Face centred central composite design (FCCCD) that underwent thorough physicochemical and mechanical assessment. Overall, robust rapidly disintegrating solid eye drops were produced. Fastest disintegration time was noted to be 0.200s. Drug content ranged from 79-96%. An improved permeation of formulations compared to a pure drug dispersion was seen. Mathematical modeling was conducted for better insight into the behavior of the device on the eye surface. Statistical analysis through constraint optimization yielded a single optimal formulation. Thermal and molecular transition analysis showed congruent findings with no incompatibility between components. Combinatory surface morphology and porositometric studies confirmed the presence of interconnecting pores across the matrix surface. Drug release kinetic evaluation predicted that best model fit was first-order release. Ocular irritancy studies by means of the HET-CAM test indicated that both drug-loaded and drug-free eye drops had an irritation score of 0 with the inference of good tolerability. Ex vivo permeation across excised rabbit cornea showed an improved steady state drug flux (0.00052mg.cm-2.min-1) and permeability co-efficient (1.7x10-4cm.min-1) for the optimized device compared to pure drug and a marketed eye drop preparation. In vivo analysis was conducted on the rabbit model with insertion of the device into the ocular cul-de-sac. Subsequently, ultra performance liquid chromatography (UPLC) analysis of the aspirated aqueous humour for model drug timolol maleate detection was conducted. The device demonstrated improved drug levels (Cmax = 3ug/mL) in comparison to commercial eye drops (Cmax = 1.97ug/mL) and was well tolerated. Level A point-to-point IVIVC plots indicated a R2 value of 0.84. This served to imply that the in vitro dissolution data can be compared to and may serve as a surrogate to that of in vivo pK data. Histopathological assessment on the enucleated eye ball confirmed the lack of noxious effects of the device on ocular tissue. From this study, the solid eye drop device was concluded to be safe as a drug delivery system for the anterior eye. Looking toward innovative trends and modifications, a bi-layered solid eye drop system with enhanced permeability capabilities employing low molecular weight chitosan was further fabricated for preliminary investigation.

by King Yeung Wong.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1999.
Includes bibliographical references (leaves 143-148).
Abstracts in English and Chinese.
Title Page --- p.i
Acknowledgement --- p.ii
List of Abbreviations --- p.iii
Table of contents --- p.v
Abstract --- p.viii
Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Liver diseases --- p.1
Chapter 1.2 --- Current treatments of liver diseases --- p.3
Chapter 1.3 --- Schizandrae --- p.5
Chapter 1.3.1 --- Chemistry of Schizandrae (Wuweizi) --- p.6
Chapter 1.3.2 --- Pharmacology of Wuweizi --- p.8
Chapter 1.3.2.1 --- Hepato-protective effect of Wuweizi --- p.9
Chapter 1.3.3 --- Toxicology and side-effects of Wuweizi --- p.11
Chapter 1.4 --- Carbon tetrachloride (CC14) intoxication --- p.12
Chapter 1.5 --- Hepatic drug metabolism: essential factors --- p.13
Chapter 1.6 --- Aim --- p.14
Chapter 2 --- Phase I metabolism --- p.15
Chapter 2.1 --- Introduction --- p.15
Chapter 2.2 --- Materials and Methods --- p.18
Chapter 2.2.1 --- Animals --- p.18
Chapter 2.2.2 --- Chemicals --- p.18
Chapter 2.2.3 --- Instruments --- p.19
Chapter 2.2.4 --- Preparation of Schizandra seed extract --- p.19
Chapter 2.2.5 --- Animal model of liver damages --- p.20
Chapter 2.2.6 --- Evaluation of protective effect of Schizandra extract --- p.22
Chapter 2.2.7 --- Evaluation of healing effect of Schizandra extract --- p.24
Chapter 2.2.8 --- Extraction of antipyrine from blood and urine --- p.26
Chapter 2.2.9 --- TLC method for quantitative analysis of antipyrine --- p.26
Chapter 2.2.10 --- Analysis of pharmacokinetic parameters of antipyrine --- p.27
Chapter 2.2.11 --- Statistical analysis --- p.28
Chapter 2.3 --- Results --- p.30
Chapter 2.3.1 --- Effect of CCI4 and Schizandra seed extract on antipyrine metabolism --- p.30
Chapter 2.4 --- Discussion --- p.41
Chapter 3 --- Phase II metabolism --- p.44
Chapter 3.1 --- Introduction --- p.44
Chapter 3.2 --- Materials and Methods --- p.46
Chapter 3.2.1 --- Chemicals --- p.46
Chapter 3.2.2 --- Preparation of Schizandra extract --- p.46
Chapter 3.2.3 --- Preparation of Salicylamide solution (for injection) --- p.47
Chapter 3.2.4 --- Preparation of 2，4-dinitrophenylhydrazine solution --- p.47
Chapter 3.2.5 --- Animal groups --- p.47
Chapter 3.2.6 --- Animal model of liver damage --- p.48
Chapter 3.2.7 --- Evaluation of the hepato-protective effect of Schizandra extract --- p.49
Chapter 3.2.8 --- Determination of serum glutamate pyruvate transaminase (SGPT/ALT) and serum glutamate oxaloacetate transaminase (SGOT/AST) --- p.50
Chapter 3.2.9 --- Salicylamide adminstration and plasma collection --- p.51
Chapter 3.2.10 --- Procession of plasma and urine samples --- p.52
Chapter 3.2.11 --- HPLC Analysis --- p.54
Chapter 3.2.12 --- Preparation of liver microsomes --- p.55
Chapter 3.2.13 --- Determination of cytochrome P450 --- p.56
Chapter 3.2.14 --- Determination of protein content of the liver microsomes --- p.57
Chapter 3.2.15 --- Data Analysis --- p.58
Chapter 3.2.16 --- Statistical Analysis --- p.58
Chapter 3.3 --- Results --- p.60
Chapter 3.3.1 --- Liver enzyme levels --- p.60
Chapter 3.3.2 --- Phase II metabolism profile of salicylamide --- p.61
Chapter 3.3.3 --- Cytochrome P450 content of liver --- p.64
Chapter 3.4 --- Discussion --- p.65
Chapter 3.4.1 --- Liver enzyme assay --- p.65
Chapter 3.4.2 --- Cytochrome P450 activity --- p.67
Chapter 3.4.3 --- Hepatic metabolism of salicylamide --- p.68
Chapter 3.4.4 --- Effect of CC14 intoxication on Phase II metabolism --- p.71
Chapter 3.4.5 --- Wuweizi actions on Phase II metabolism --- p.73
Chapter 4 --- Protein binding --- p.102
Chapter 4.1 --- Introduction --- p.102
Chapter 4.2 --- Materials and Methods --- p.104
Chapter 4.2.1 --- Chemicals --- p.104
Chapter 4.2.2 --- Instruments --- p.105
Chapter 4.2.3 --- Preparation of Warfarin sodium solution --- p.105
Chapter 4.2.4 --- Animal groups --- p.106
Chapter 4.2.5 --- Equilibrium dialysis --- p.106
Chapter 4.2.5.1 --- Equilibration time --- p.106
Chapter 4.2.5.2 --- Equilibrium dialysis of different warfarin concentration --- p.107
Chapter 4.2.6 --- High performance liquid chromatography analysis of warfarin --- p.108
Chapter 4.2.7 --- Calibration curve --- p.109
Chapter 4.3 --- Results --- p.111
Chapter 4.3.1 --- Equilibriation time --- p.111
Chapter 4.3.2 --- Calibration curve --- p.111
Chapter 4.3.3 --- Free concentration of warfarin --- p.112
Chapter 4.4 --- Discussion --- p.114
Chapter 4.4.1 --- Effect of CCl4 intoxication on free percentage of warfarin --- p.114
Chapter 4.4.2 --- Effcct of wuweizi cxtract on free percentage of warfarin --- p.115
Chapter 4.4.2.1 --- Depletion of plasma albumin concentration --- p.116
Chapter 4.4.2.2 --- Displacement of warfarin by WWZ extract --- p.117
Chapter 4.4.3 --- Concentration dependent protein binding --- p.118
Chapter 5 --- Hepatic blood flow --- p.124
Chapter 5.1 --- Introduction --- p.124
Chapter 5.2 --- Materials and Methods --- p.126
Chapter 5.2.1 --- Chemicals....: --- p.126
Chapter 5.2.2 --- Instruments --- p.126
Chapter 5.2.3 --- Preparation of indocyanine green (ICG) solution --- p.126
Chapter 5.2.4 --- Preparation of Schizandra seed extract --- p.127
Chapter 5.2.5 --- Animals groups --- p.127
Chapter 5.2.6 --- Animal model of liver damage --- p.128
Chapter 5.2.7 --- Evaluation of hepato-protective effect of Schizandra extract --- p.129
Chapter 5.2.8 --- Evaluation of healing effect of Schizandra extract --- p.129
Chapter 5.2.9 --- Quantitative analysis of ICG in plasma by UV spectroscopy --- p.130
Chapter 5.2.10 --- Analysis of pharmacokinetic parameters of ICG --- p.131
Chapter 5.2.11 --- Statistical analysis --- p.132
Chapter 5.3 --- Results --- p.133
Chapter 5.4 --- Discussion --- p.135
Chapter 5.4.1 --- Effect of CCl4 intoxication on hepatic blood flow --- p.135
Chapter 5.4.2 --- Effect of WWZ pretreatment on hepatic blood flow --- p.135
Chapter 5.4.3 --- Effect of WWZ healing on hepatic blood flow --- p.136
Chapter 6 --- General conclusion --- p.139
Significance of the study --- p.141
References --- p.143

Wang Deqing.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (p. 236-277).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

Conclusion 1. The results of the research partly supported hypothesis 1a. There was a significant reduction in both endothelium-dependent arterial relaxation following a surgical menopause. The results of the research partly supported hypothesis 1b. There was a significant reduction in endothelium-dependent arterial relaxation but no significant effect on endothelium-independent arterial relaxation.
Conclusion 2. The results of the research partly supported hypothesis 2a. The addition of unopposed oestrogen significantly improved endothelium-dependent but not endothelium-independent arterial relaxation. The results of the research supported hypothesis 2b. The addition of oestradiol combined with progestogen (norethisterone acetate) reversed the reduction in arterial relaxation caused by a surgical menopause. The results of the research partly supported hypothesis 2c. The addition of tibolone reversed the reduction endothelium-dependent but not endothelium-independent arterial relaxation. The results of the research partly supported hypothesis 2d. The addition of oestradiol combined with a progestogen (norethisterone acetate) reversed the reduction in endothelium-dependent but not endothelium-independent arterial relaxation.
Conclusion 3. The results of the research partly supported hypothesis 3a. Endothelium-dependent arterial relaxation but no endothelium-independent arterial relaxation was improved after the addition of menopausal hormone therapy using oestrogen combined with a progestogen in a continuous manner. The results of the research did not support hypothesis 3b. Neither endothelium-dependent arterial relaxation nor the endothelium-independent arterial relaxation was improved by cyclical menopausal HT.
Conclusion 4. The results of the research did not support hypothesis 4. The addition of menopausal hormone therapy using combined oestrogen with progestogen did not improve arterial relaxation in postmenopausal women with established coronary heart disease.
Hypothesis 2. This hypothesis examined three different types of commonly used menopausal HT. That unopposed oestrogen (2a), oestrogen combined with a progestogen (2b and 2d) or a synthetic steriod that has oestrogenic, progestogenic as well as androgenic activity (tibolone, 2c), reverse the reduction in arterial relaxation following menopause in Hong Kong Chinese women.
Hypothesis 3. That menopausal hormone therapy using oestrogen combined with progestogen given in either continuous (3a) or cyclical (3b) regimens improves arterial relaxation in postmenopausal Hong Kong Chinese women.
Hypothesis 4. That menopausal hormone therapy using combined oestrogen with progestogen improves arterial relaxation in postmenopausal Hong Kong Chinese women with established coronary heart disease.
Menopausal HT can in general at least partially reverse changes in arterial relaxation in postmenopausal women. Different types of menopausal HT exhibit different effects on arterial relaxation. In healthy vessels, menopause HT mainly reverses the endothelium-dependent vascular effect, but it remains unclear how menopausal HT affects the endothelium-independent vascular effect. However, with established coronary heart disease, menopausal HT cannot reverse the changes in vascular reactivity.
Summary. Menopause results in a reduction in arterial relaxation. However, GnRHa temporarily induced menopause in young women, the endothelium-independent vasodilatation was not impaired. This difference can be partly explained by the difference in age as vascular reactivity is age dependent. Secondly, GnRHa works with an initial phase of increase in oestrogen production resulting in a shorter duration of hypo-oestrogenism resulting in the lack of impairment on endothelium-independent vasodilatation.
This thesis tested the following hypotheses: Hypothesis 1. That vascular reactivity decreases after the menopause as shown in premenopausal Hong Kong Chinese women with either a surgical (1a) or a medically induced (1b) menopause.
This thesis will examine the effects of menopause and menopausal HT on arterial reactivity which is an indirect measurement of vascular function. Previous studies have shown that oestrogen is a potent coronary artery vasodilator, and this effect may be mediated via both endothelium-dependent and endothelium-independent mechanisms. One method of assessing vascular reactivity is to use ultrasound measurement of changes in brachial artery diameter in response to certain stimuli. Using this technique, changes in both endothelium-dependent and endothelium-independent vasodilatation can be measured. Increased rather than decreased arterial relaxation after stimulus can be viewed as a favourable response.
Yim, So-fan.
Adviser: C. J. Haines.
Source: Dissertation Abstracts International, Volume: 68-09, Section: B, page: 5873.
Thesis (M.D.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (p. 159-194).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
School code: 1307.

Thesis (M.Pharm.)--University of the Witwatersrand, Faculty of Health Sciences, 2013.
Sexually transmitted infections (STIs) are a global concern and more specifically southern Africa has seen a tremendous upsurge in infection rates. KwaZulu-Natal is the province found to have the highest Human Immunodeficiency Virus and STI infection rates. From an ethnobotanical study conducted specifically in northern Maputaland (Mabibi, Tshongwe, Mseleni and Mbazwana), it was found that the lay people most often used plants in various combinations for the treatment of STI related symptoms. The use of these plant combinations were thus antimicrobially investigated and the toxicity properties determined. The dichloromethane: methanol (organic) and aqueous extracts were prepared for each plant in situ using collected ground dried plant material. The plants (individually and in combination) were investigated for toxic potential using the 3-[4,5-dimethyl-2-thiazol-yl]-2,5-diphenyl-2H-tetrazolium bromide (MTT) cellular viability assay on the human kidney epithelial (Graham) cell line. The antimicrobial activities for each sample, as well as for each combination, were then further investigated using the minimum inhibitory concentration (MIC) assay. The six STI pathogens investigated in this study were Candida albicans (ATCC 10321), Ureaplasma urealyticum (clinical strain), Oligella ureolytica (ATCC 43534), Gardnerella vaginalis (ATCC 14018), Trichomonas vaginalis (clinical strain) and Neisseria gonnorhoeae (ATCC 19424).

Chan Chun-pong.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (leaves 171-176).
Abstracts in English and Chinese.
Acknowledgements --- p.i
Abstract --- p.ii
摘要 --- p.iii
Abbreviation --- p.iv
Table of contents --- p.v
List of figures --- p.xi
List of tables --- p.xvi
Chapter Chapter 1 --- introduction
Chapter 1.1 --- Traditional Chinese medicines (TCMs) --- p.2
Chapter 1.2 --- Liver disorders in Asia Pacific region --- p.3
Chapter 1.3 --- Classification of liver disorders --- p.7
Chapter 1.3.1 --- Hepatitis --- p.8
Chapter 1.3.1.1 --- Hepatitis A virus infection --- p.8
Chapter 1.3.1.2 --- Hepatitis B virus infection --- p.9
Chapter 1.3.1.3 --- Hepatitis C virus infection --- p.11
Chapter 1.3.1.4 --- Hepatitis D virus infection --- p.12
Chapter 1.3.1.5 --- Hepatitis E virus infection --- p.12
Chapter 1.3.2 --- Cancer of the liver --- p.13
Chapter 1.3.2.1 --- Hepatocellular carcinoma --- p.13
Chapter 1.3.2.2 --- Cholangiocarcinoma --- p.14
Chapter 1.3.2.3 --- Metastatic liver cancer --- p.14
Chapter 1.4 --- Conventional treatment of liver disorders --- p.14
Chapter 1.5 --- Role of traditional Chinese medicines in hepatoprotective functions --- p.16
Chapter 1.5.1 --- Abri Herba (Abrus Cantoniensis Hance) --- p.17
Chapter 1.5.2 --- Rhizoma Coptidis (Coptidis chinensis Franch) --- p.18
Chapter 1.5.3 --- Fructus Forsythia (Forsythia suspense (Thunb) Vahl) --- p.22
Chapter 1.6 --- Molecular studies of hepatoprotective effects of TCMs --- p.26
Chapter 1.6.1 --- Roles of detoxofication enzymes in hepatoprotection --- p.27
Chapter 1.6.2 --- Studies of growth-related genes in cell cycle control --- p.29
Chapter 1.7 --- Aims of project --- p.32
Chapter 1.8 --- Application of the project --- p.33
Chapter Chapter 2 --- Methods and materials --- p.34
Chapter 2.1 --- Screening of traditional Chinese medicines --- p.35
Chapter 2.2 --- Preparation of TCMs --- p.35
Chapter 2.2.1 --- Preparation of aqueous extracts of TCMs --- p.35
Chapter 2.2.2 --- Preparation of active components of TCMs --- p.36
Chapter 2.3 --- In vitro assays --- p.40
Chapter 2.3.1 --- Cell culture --- p.40
Chapter 2.3.2 --- Cytotoxicity test --- p.40
Chapter 2.4 --- Screening of expressed gene induced by TCMs --- p.41
Chapter 2.4.1 --- RNA preparation --- p.41
Chapter 2.4.2 --- cDNA array hybridization --- p.42
Chapter 2.4.3 --- Reverse Transcription --- p.43
Chapter 2.5 --- Confirmation of expressed genes induced by TCMs --- p.44
Chapter 2.5.1 --- Semi-quantitative PCR analysis --- p.44
Chapter 2.5.2 --- Northern blot analysis --- p.46
Chapter 2.6 --- Studies of effects of TCMs in gene expression --- p.47
Chapter 2.6.1 --- Dosage-course study --- p.47
Chapter 2.6.2 --- Time-course study --- p.48
Chapter Chapter 3 --- Results --- p.50
Chapter 3.1 --- "Cytotoxicity test of A.H., R.C. and F.F" --- p.51
Chapter 3.2 --- "Molecular screening of expressed gene induced by A.H., R.C., F.F" --- p.58
Chapter 3.3 --- Confirmation of expressed gene using semi-quantitative RT- PCR --- p.70
Chapter 3.3.1 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.70
Chapter 3.3.2 --- Dosage-course and time-course studies of R.C. using RT- PCR --- p.94
Chapter 3.3.3 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.113
Chapter 3.4 --- Confirmation of expressed gene using northern blot anaylsis --- p.118
Chapter 3.4.1 --- Dosage-course and time-course studies of effects of A.H. and L- abrine in Northern blot analysis --- p.118
Chapter 3.4.2 --- Dosage-course and time-course studies of effects of R.C. and berberine in Northern blot analysis --- p.129
Chapter 3.4.3 --- Dosage-course and time-course studies of effects of F.Fin Northern blot analysis --- p.147
Chapter Chapter 4 --- Discussion --- p.152
Chapter 4.1 --- "Roles of A.H., R.C. and F.F. in treatment and prevention of liver disorders" --- p.153
Chapter 4.2 --- "Cytotoxicity effect A.H., R.C., and F.F. in liver cells" --- p.153
Chapter 4.3 --- Effects of herbal medicines on the transcription of mRNA in liver cells --- p.155
Chapter 4.3.1 --- Effects of treatment of A.H. in liver at transcriptional level … --- p.155
Chapter 4.3.2 --- Effects of treatment of R.C. in liver at transcriptional level … --- p.156
Chapter 4.3.3 --- Effects of treatment of R.C. in liver at transcriptional level --- p.157
Chapter 4.4 --- Comparison of results of RT-PCR and Northern blot analysis --- p.157
Chapter 4.4.1 --- Comparison of the effects of time and dosage-course studies of DTD expression induced by A.H. and L-abrine --- p.157
Chapter 4.4.2 --- Comparison of the effects of time and dosage-course studies of p21;cip;waf1 expression induced by A.H. and L-abrine --- p.158
Chapter 4.4.3 --- Comparison of the effects of time and dosage-course studies of c-myc responsive protein; rcl expression induced by R.C. and berberine --- p.159
Chapter 4.4.4 --- Comparison of the effects of time and dosage-course studies of GST Ya expression induced by R.C. and berberine --- p.160
Chapter 4.4.5 --- Comparison of the effects of time and dosage-course studies of GST 7-7 expression induced by F.F --- p.160
Chapter 4.5 --- Biochemical significance of genes induced by hepatoprotective TCMs --- p.161
Chapter 4.5.1 --- Roles of significant expression of detoxifying enzymes induced by TCMs in liver cells --- p.161
Chapter 4.5.2 --- Roles of induction of growth-related c-myc responsive protein; rcl in R.C. treated liver cells --- p.167
Chapter 4.5.3 --- Roles of increased p21;cip;waf1 expression in A.H. treated liver cells --- p.168
Chapter 4.6 --- Conclusion --- p.169

Chan Hoi Yun.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 131-144).
Abstracts in English and Chinese.
DECLARATION --- p.i
ACKNOWLEDGMENTS --- p.ii
ABBREVIATIONS --- p.iii
ABSTRACT IN ENGLISH --- p.v
ABSTRACT IN CHINESE --- p.viii
CONTENTS --- p.xi
Chapter Chapter 1 --- Introduction
Chapter 1.1. --- Steroid Hormones --- p.1
Chapter 1.1.1. --- Synthesis of estrogens and progesterone --- p.1
Chapter 1.2. --- Cellular Mechanisms of Female Steroid Hormones --- p.5
Chapter 1.2.1. --- Genomic actions of female steroid hormones --- p.5
Chapter 1.2.2. --- Non-genomic actions of female steroid hormones --- p.7
Chapter 1.2.3. --- Estrogen antagonists --- p.7
Chapter 1.2.3.1. --- Classification of estrogen antagonists --- p.7
Chapter 1.2.3.2. --- Mechanisms of estrogen antagonists --- p.9
Chapter 1.3. --- Chronic (genomic) Effects of 17β-Estradiol and Progesterone --- p.10
Chapter 1.3.1. --- Effects of lipid metabolism --- p.10
Chapter 1.3.2. --- Effects on cell proliferation --- p.11
Chapter 1.3.3. --- Effects on endothelial cells --- p.12
Chapter 1.4. --- Acute Effects of 17β-Estradiol and Progesterone --- p.13
Chapter 1.4.1. --- Role of endothelium in 17β-estradiol or progesterone Relaxation --- p.13
Chapter 1.4.2. --- Involvement of plasma membrane estrogen receptors --- p.14
Chapter 1.4.3. --- Role of Ca2+ and K+ channel in estrogen relaxation --- p.14
Chapter 1.4.4. --- Interaction with vasoconstrictors --- p.15
Chapter 1.4.5. --- Interaction with endothelium-dependent dilators --- p.16
Chapter 1.4.6. --- Interaction with adrenergic response --- p.17
Chapter 1.5. --- Clinical Studies --- p.19
Chapter 1.6. --- Therapeutic Values of Estrogen and Progesterone --- p.20
Chapter 1.7. --- Objectives of the Present Study --- p.22
Chapter Chapter 2 --- Method and Materials
Chapter 2.1. --- Tissue Preparation --- p.25
Chapter 2.1.1. --- "Preparation of the rat aorta, mesenteric artery and carotid Artery" --- p.25
Chapter 2.1.2. --- Removal of the functional endothelium --- p.26
Chapter 2.2. --- Organ Bath Set-up --- p.26
Chapter 2.3. --- Force Measurement --- p.28
Chapter 2.3.1. --- Vascular action of 17β-estradiol and progesterone --- p.29
Chapter 2.3.1.1. --- Role of endothelium/nitric oxide in 17β-estradiol- or progesterone-induced relaxation --- p.29
Chapter 2.3.1.2. --- Role of inducible nitric oxide in progesterone-induced relaxation --- p.30
Chapter 2.3.1.3. --- Effect of estrogen receptor inhibitor on 17β-estradiol- induced relaxation --- p.30
Chapter 2.3.1.4. --- Interaction between progesterone and 17β-estradiol --- p.31
Chapter 2.3.1.5. --- Effect of 17β-estradiol on protein kinase C-mediated contraction --- p.31
Chapter 2.3.1.6. --- Synergistic interaction between β-adrenoceptor agonists and 17β-estradiol --- p.32
Chapter 2.4. --- Porcine Coronary Artery Experiments --- p.33
Chapter 2.4.1. --- Vessel preparation --- p.33
Chapter 2.4.2. --- Force measurement --- p.33
Chapter 2.4.3. --- Experimental protocol --- p.34
Chapter 2.4.3.1. --- Effect of physiological level of 17β-estradiol on β- adrenoceptor agonist-induced relaxation --- p.34
Chapter 2.4.3.2. --- Effect of physiological level of 17β-estradiol on phosphodiesterase inhibitor-induced relaxation --- p.34
Chapter 2.5. --- Ovariectomy --- p.35
Chapter 2.5.1. --- Method of ovariectomy --- p.35
Chapter 2.5.2. --- Preparation of blood vessels --- p.36
Chapter 2.5.3. --- Experimental protocols --- p.38
Chapter 2.5.3.1. --- Effect of ovariectomy on contractility of rat carotid arteries --- p.38
Chapter 2.5.3.2. --- Effect of ovariectomy on relaxation of rat carotid arteries --- p.38
Chapter 2.6. --- Chemicals and Solutions --- p.39
Chapter 2.7. --- Statistical Analysis --- p.42
Chapter Chapter 3 --- Results
Chapter 3.1. --- Role of Endothelium/Nitric Oxide in 17β-Estradiol- and Progesterone-induced Relaxations --- p.43
Chapter 3.1.1. --- Relaxant response of 17β-estradiol --- p.43
Chapter 3.1.2. --- Effects of inhibitors of nitric oxide activity on 17β- estradiol-induced relaxation --- p.46
Chapter 3.1.3. --- Relaxant response of progesterone --- p.46
Chapter 3.1.4. --- Effects of inhibitors of nitric oxide activity on progesterone-induced relaxation --- p.50
Chapter 3.2. --- Effect of Estrogen Receptor Inhibitor on 17β-Estradiol- induced Relaxation --- p.56
Chapter 3.3. --- Interaction between Progesterone and 17β-Estradiol --- p.56
Chapter 3.4. --- Effect of Female Sex Steroid Hormones on Protein Kinase C-mediated Contraction --- p.59
Chapter 3.4.1. --- Effect of 17β-estradiol on phorbol ester-induced contraction --- p.59
Chapter 3.4.2. --- Effect of progesterone on phorbol ester-induced contraction --- p.59
Chapter 3.5. --- Effects of β-adrenoceptor Agonists on 17β-Estradiol- induced Relaxations --- p.62
Chapter 3.5.1. --- Effect of isoproterenol on 17β-estradiol-induced relaxation --- p.62
Chapter 3.5.2. --- Role of endothelium/nitric oxide on the isoproterenol potentiation of 17β-estradiol-induced relaxation --- p.63
Chapter 3.5.3. --- Role of cyclic AMP on isoproterenol-enhancement of 17β- estradiol-induced relaxation --- p.69
Chapter 3.5.4. --- Effects of β-adrenoceptor antagonists --- p.69
Chapter 3.6. --- Effects of Physiological Concentration of 17β-EstradioI onβ-adrenoceptor Agonists-induced Relaxationsin Porcine Coronary Artery --- p.77
Chapter 3.6.1. --- Effect of 17β-estradiol on isoproterenol-induced relaxations --- p.77
Chapter 3.6.2. --- Effect of 17β-estradiol on fenoterol-induced relaxations --- p.11
Chapter 3.6.3. --- Effect of 17β-estradiol on dobutamine-induced relaxations --- p.81
Chapter 3.6.4. --- Effect of 17β-estradiol on IBMX-induced relaxation --- p.86
Chapter 3.7. --- Effect of Ovariectomy on the Vascualr Reactivity --- p.88
Chapter 3.7.1. --- Effect of ovariectomy on the contractile activity of rat carotid artery --- p.88
Chapter 3.7.1.1. --- Effect of ovariectomy on phenylephrine-induced contraction --- p.88
Chapter 3.7.1.2. --- Effect of ovariectomy on U46619-induced contraction --- p.96
Chapter 3.7.1.3. --- Effect of ovariectomy on high K+- induced contraction --- p.102
Chapter 3.7.1.4. --- Effect of ovariectomy on acetylcholine-induced relaxation --- p.106
Chapter Chapter 4 --- Discussions
Chapter 4.1. --- Role of Endothelium/Nitric oxide in 17β-Estradiol- and Progesterone-induced Relaxations --- p.110
Chapter 4.2. --- Effect of Estrogen Receptor Inhibitor on 17β-Estradiol- induced Relaxation --- p.113
Chapter 4.3. --- Interaction between Progesterone and 17β-Estradiol --- p.114
Chapter 4.4. --- Effects of Female Sex Steroid Hormones on Protein Kinase C-mediated Contraction --- p.115
Chapter 4.5. --- Effects of β-Adrenoceptor Agonists on 17β-Estradiol- induced Relaxations --- p.116
Chapter 4.6. --- Effects of 17β-Estradiol on β-Adrenoceptor Agonists- induced Relaxations in Porcine Coronary Artery --- p.121
Chapter 4.7. --- Effect of Ovariectomy on the Vascular Reactivity --- p.125
Chapter 4.8. --- Conclusions --- p.129
References --- p.131
Publications --- p.145

Ho Hing Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 140-156).
Abstracts in English and Chinese.
Chapter Chapter 1 --- General introduction
Chapter 1.1 --- History of soybean --- p.1
Chapter 1.2 --- Health benefits of soybean --- p.2
Chapter 1.3 --- Introduction to flavonoids --- p.2
Chapter 1.4 --- Bioavailability of flavonoids from foods --- p.3
Chapter 1.5 --- Pharmacological effects of flavonoids and their glycosides --- p.4
Chapter 1.5.1 --- Anticarcinogenic activity --- p.4
Chapter 1.5.2 --- Antioxidative activity --- p.7
Chapter 1.5.3 --- Cardioprotective activity --- p.9
Chapter 1.5.4 --- Osteoprotective activity --- p.10
Chapter 1.5.5 --- Neuroprotective activity --- p.12
Chapter 1.5.6 --- Antiangiogenic activity --- p.12
Chapter 1.6 --- Soy leaves --- p.13
Chapter Chapter 2 --- Isolation and purification of kaempferol glycosides and genistin in soy leaves
Chapter 2.1 --- Introduction --- p.14
Chapter 2.2 --- Objectives --- p.15
Chapter 2.3 --- Materials and Methods --- p.16
Chapter 2.3.1 --- Extraction and isolation --- p.16
Chapter 2.3.1.1 --- Preparation of soy leaves butanol extract --- p.16
Chapter 2.3.1.2 --- Preparation of kaempferol glycosides from soy leaves butanol extract --- p.16
Chapter 2.3.2 --- High performance liquid chromatography (HPLC) analysis --- p.19
Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.19
Chapter 2.3.2.2 --- HPLC analysis --- p.19
Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.23
Chapter 2.3.2.4 --- Change in flavonoids and their glycosides in soy leaves --- p.23
Chapter 2.4 --- Results --- p.24
Chapter 2.4.1 --- Compound 1 --- p.24
Chapter 2.4.2 --- Compound 2 --- p.24
Chapter 2.4.3 --- Compound 3 --- p.25
Chapter 2.4.4 --- Compound 4 --- p.25
Chapter 2.4.5 --- Compound 5 --- p.25
Chapter 2.4.6 --- Compound 6 --- p.26
Chapter 2.4.7 --- Quantification of flavonoids in soybean and soy leaves --- p.32
Chapter 2.4.8 --- Age-dependent changes in flavonoids and their glycosides --- p.32
Chapter 2.5 --- Discussion --- p.35
Chapter 2.5.1 --- Compound 1 --- p.35
Chapter 2.5.2 --- Compound 2 --- p.35
Chapter 2.5.3 --- Compound 3 --- p.37
Chapter 2.5.4 --- Compound 4 --- p.38
Chapter 2.5.5 --- Compound 5 --- p.39
Chapter 2.5.6 --- Compound 6 --- p.40
Chapter 2.5.7 --- Age-dependent changes in flavonoids and their glycosides --- p.40
Chapter Chapter 3 --- Hypolipidemic effects of soy leaves in hamsters
Chapter 3.1 --- Introduction --- p.41
Chapter 3.1.1 --- Different lipoproteins and their functions --- p.41
Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.42
Chapter 3.1.3 --- Animal model --- p.43
Chapter 3.2 --- Objectives --- p.44
Chapter 3.3 --- Materials and Methods --- p.45
Chapter 3.3.1 --- Animals --- p.46
Chapter 3.3.2 --- Serum lipid and lipoprotein determinations --- p.46
Chapter 3.3.3 --- Determination of cholesterol in the liver and adipose tissue --- p.46
Chapter 3.3.4 --- Extraction of neutral and acidic sterols from fecal samples --- p.49
Chapter 3.3.4.1 --- Determination of neutral sterols --- p.49
Chapter 3.3.4.2 --- Determination of acidic sterols --- p.50
Chapter 3.3.4.3 --- GLC analysis of neutral and acidic sterols --- p.51
Chapter 3.3.5 --- Statistics --- p.51
Chapter 3.4 --- Results --- p.54
Chapter 3.4.1 --- Growth and food intake --- p.54
Chapter 3.4.2 --- "Effects of SLP and SLEE supplementation on serum triacylglycerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C)" --- p.54
Chapter 3.4.3 --- Effects ofSLP and SLEE supplementation on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.55
Chapter 3.4.4 --- Effects of SLP amd SLEE supplementations on concentration of hepatic cholesterol --- p.58
Chapter 3.4.5 --- Effects of SLP and SLEE supplementations on perirenal adipose tissue cholesterol --- p.58
Chapter 3.4.6 --- Effects of SLP and SLEE supplementations on fecal neutral and acidic sterols --- p.61
Chapter 3.5 --- Discussion --- p.64
Chapter Chapter 4 --- Effects of soy leaves and its flavonoid glycosides on haemolysis and on LDL oxidation
Chapter 4.1 --- Introduction --- p.67
Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.68
Chapter 4.1.2 --- LDL oxidation --- p.70
Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.71
Chapter 4.1.4 --- Antioxidant and LDL oxidation --- p.74
Chapter 4.2 --- Objective --- p.75
Chapter 4.3 --- Materials and methods --- p.76
Chapter 4.3.1 --- Isolation of LDL from human serum --- p.76
Chapter 4.3.2 --- LDL oxidation --- p.77
Chapter 4.3.3 --- Determine the formation of thiobarbituric acid-reactive substances (TBARS) --- p.77
Chapter 4.3.4 --- Assay for erythrocyte haemolysis --- p.78
Chapter 4.3.5 --- Statistics --- p.79
Chapter 4.4 --- Results --- p.80
Chapter 4.4.1 --- Effects of three different soy leaves extracts and flavonoid glycosides on LDL oxidation --- p.80
Chapter 4.4.2 --- Effects of three soy leaves extracts and flavonoid glycosides on erythrocyte haemolysis --- p.80
Chapter 4.5 --- Discussion --- p.85
Chapter Chapter 5 --- Relaxing effects of soy leaves and its flavonoids
Chapter 5.1 --- Introduction --- p.89
Chapter 5.1.1 --- Smooth muscle contraction --- p.90
Chapter 5.1.1.1 --- Sliding filament mechanism --- p.91
Chapter 5.1.2 --- Intracellular mechanisms involved in the regulation of smooth muscle contraction --- p.92
Chapter 5.1.2.1 --- Voltage-gated Ca2+ channels --- p.92
Chapter 5.1.2.2 --- Protein kinase C (PKC) mediated smooth muscle contraction --- p.93
Chapter 5.1.2.3 --- Thromboxane A2 receptor-mediated calcium channel --- p.94
Chapter 5.2 --- Objectives --- p.96
Chapter 5.3 --- Materials and methods --- p.97
Chapter 5.3.1 --- Drugs preparation --- p.97
Chapter 5.3.2 --- Vessel preparation --- p.97
Chapter 5.3.3 --- Contraction experiments --- p.99
Chapter 5.3.3.1 --- Relaxant responses of soy leaves butanol extract on the contraction induced by different constrictors --- p.99
Chapter 5.3.3.2 --- Relaxant responses of soy leaves butanol extract on U46619 and PGF2a- induced contraction --- p.99
Chapter 5.3.3.3 --- "Relaxant responses of genistein, genistin and the kaempferol glycosides on U46619-induced contraction" --- p.100
Chapter 5.3.4 --- Statistics --- p.100
Chapter 5.4 --- Results --- p.102
Chapter 5.4.1 --- Effect of soy leaves butanol extract --- p.102
Chapter 5.4.2 --- Role of endothelium in extract-induced relaxation --- p.102
Chapter 5.4.3 --- Effect of the soy leaves butanol extract on contractile response to prostaglandins --- p.103
Chapter 5.4.4 --- Effects of kaempferol glycosides and kaempferol --- p.111
Chapter 5.4.5 --- Effects of genistein and genistin --- p.111
Chapter 5.5 --- Discussion --- p.118
Chapter Chapter 6 --- Effect of soy leaves on mammary tumor
Chapter 6.1 --- Introduction --- p.123
Chapter 6.1.1 --- Carcinogenesis --- p.123
Chapter 6.1.1.1 --- In itiation --- p.124
Chapter 6.1.1.2 --- Promotion --- p.124
Chapter 6.1.1.3 --- Progression --- p.125
Chapter 6.2 --- Objective --- p.126
Chapter 6.3 --- Materials and methods --- p.127
Chapter 6.3.1 --- Animal --- p.127
Chapter 6.3.2 --- Determination of estrus cycle --- p.128
Chapter 6.3.3 --- Statistics --- p.129
Chapter 6.4 --- Results --- p.131
Chapter 6.4.1 --- Incident rate of tumor induction --- p.131
Chapter 6.4.2 --- Number of tumor induced --- p.131
Chapter 6.5 --- Discussion --- p.136
Chapter Chapter 7 --- Conclusions --- p.136
References --- p.140

Keung Hoi Yee.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 151-174).
Abstracts in English and Chinese.
Acknowledgement --- p.I
Abstract --- p.II
摘要 --- p.V
Content --- p.VII
List of tables --- p.XIII
List of figures --- p.XIV
Abbreviations --- p.XVII
Chapter Chapter 1 --- General Introduction --- p.1
Chapter 1.1 --- Biology of Pleurotus tuber-regiun (Ptr) --- p.1
Chapter 1.1.1 --- Ptr grown in the wild --- p.1
Chapter 1.1.2 --- Cultivation of Ptr --- p.2
Chapter 1.2 --- Functional food and pharmaceutical application of Ptr sclerotium --- p.3
Chapter 1.2.1 --- Traditional food and medicinal uses of Ptr sclerotium --- p.3
Chapter 1.2.2 --- Nutritional value and chemical composition --- p.4
Chapter 1.2.3 --- Anti-tumor activity --- p.7
Chapter 1.2.4 --- Anti-viral activity --- p.8
Chapter 1.2.5 --- Immunologic function --- p.8
Chapter 1.2.6 --- Pharmaceutical application --- p.9
Chapter Chapter 2 --- Toxicological evaluation on Ptr sclerotium --- p.11
Chapter 2.1 --- Introduction --- p.11
Chapter 2.1.1 --- Toxicological concern of Ptr sclerotium --- p.11
Chapter 2.1.2 --- Toxicological study --- p.12
Chapter 2.1.3 --- Biochemical methods for toxicological evaluation --- p.14
Chapter 2.1.3.1 --- Serum enzyme activities --- p.15
Chapter 2.1.3.2 --- Other serum analytes --- p.17
Chapter 2.1.4 --- Histopathological study --- p.20
Chapter 2.1.5 --- Acute toxicity --- p.21
Chapter 2.1.6 --- Sub-acute and sub-chronic toxicity --- p.23
Chapter 2.1.7 --- Objectives --- p.26
Chapter 2.2 --- Materials and Methods --- p.27
Chapter 2.2.1 --- Sample materials and chemicals --- p.27
Chapter 2.2.2 --- Acute toxicity test --- p.27
Chapter 2.2.2.1 --- Diet and animals --- p.27
Chapter 2.2.2.2 --- Experimental design --- p.28
Chapter 2.2.2.3 --- Calculation of sclerotium intake dose --- p.29
Chapter 2.2.2.4 --- Biochemical assays --- p.30
Chapter 2.2.2.5 --- Histopathological examination --- p.31
Chapter 2.2.3 --- Sub-acute and sub-chronic toxicity tests --- p.32
Chapter 2.2.3.1 --- Diet Preparation --- p.32
Chapter 2.2.3.2 --- Experimental design --- p.32
Chapter 2.2.3.3 --- Biochemical assays --- p.36
Chapter 2.2.3.4 --- Organ weight --- p.40
Chapter 2.2.3.5 --- Histopathological examination --- p.41
Chapter 2.2.4 --- Statistical analyses --- p.41
Chapter 2.3 --- Results and Discussion --- p.42
Chapter 2.3.1 --- Acute toxicity test --- p.42
Chapter 2.3.1.1 --- Food consumption --- p.43
Chapter 2.3.1.2 --- Serum transaminase activities --- p.44
Chapter 2.3.1.3 --- Histopathology --- p.45
Chapter 2.3.1.4 --- NOAEL --- p.45
Chapter 2.3.2 --- Sub-acute toxicity test --- p.50
Chapter 2.3.2.1 --- Body weight gain --- p.50
Chapter 2.3.2.2 --- Biochemical assays --- p.51
Chapter 2.3.2.3 --- Organ per body weight and histopathology --- p.52
Chapter 2.3.2.4 --- Effects of Ptr sclerotial diets --- p.53
Chapter 2.3.3 --- Sub-chronic toxicity test --- p.59
Chapter 2.3.3.1 --- Food and energy consumption --- p.59
Chapter 2.3.3.2 --- Biochemical assays --- p.63
Chapter 2.3.3.3 --- Organ per body weight --- p.67
Chapter 2.3.3.4 --- Body weight increase --- p.75
Chapter 2.3.3.5 --- NOAEL --- p.80
Chapter 2.4 --- Summary --- p.81
Chapter Chapter 3 --- Hepatoprotection of Ptr sclerotium --- p.82
Chapter 3.1 --- Introduction --- p.82
Chapter 3.1.1 --- Hepatotoxicity --- p.82
Chapter 3.1.2 --- Potential hepatoprotection effect of Ptr sclerotium --- p.83
Chapter 3.1.3 --- Toxicity of CC14 --- p.85
Chapter 3.1.4 --- Toxicity of AFB! --- p.89
Chapter 3.1.5 --- Bioactivity of chlorophyllin --- p.92
Chapter 3.1.6 --- Comet assay --- p.93
Chapter 3.1.7 --- Objectives --- p.98
Chapter 3.2 --- Materials and Methods --- p.99
Chapter 3.2.1 --- Sample materials and chemicals --- p.99
Chapter 3.2.2 --- Curative and preventive tests of Ptr sclerotium against CCl4-induced hepatotoxicity --- p.99
Chapter 3.2.2.1 --- Animal and diets --- p.99
Chapter 3.2.2.2 --- Dose-response of CCl4 on rat model --- p.100
Chapter 3.2.2.3 --- Biochemical assays --- p.100
Chapter 3.2.2.4 --- Curative hepatoprotection test on Ptr --- p.101
Chapter 3.2.2.5 --- Preventive hepatoprotection test on Ptr --- p.101
Chapter 3.2.3 --- Preventive tests of Ptr sclerotium against AFB1-induced hepato- and geno-toxicity --- p.103
Chapter 3.2.3.1 --- Dose-response of AFB1 on rat model --- p.103
Chapter 3.2.3.2 --- Preventive test of Ptr against AFB1 --- p.103
Chapter 3.2.3.3 --- Biochemical assays --- p.105
Chapter 3.2.3.4 --- Histopathological examination --- p.105
Chapter 3.2.4 --- Comet assay --- p.106
Chapter 3.2.4.1 --- Reagent preparations --- p.106
Chapter 3.2.4.2 --- Procedures --- p.107
Chapter 3.2.5 --- Statistical analyses --- p.110
Chapter 3.3 --- Results and Discussion --- p.111
Chapter 3.3.1 --- Curative and preventive tests of Ptr sclerotium against CCl4-induced hepatotoxicity --- p.112
Chapter 3.3.1.1 --- Dose-response of CCl4 on rat model --- p.112
Chapter 3.3.1.2 --- Curative test of Ptr sclerotium against CCl4-induced hepatotoxicity --- p.116
Chapter 3.3.1.3 --- Preventive test of Ptr sclerotium against CCl4-induced hepatotoxicity --- p.121
Chapter 3.3.2 --- Preventive tests of Ptr sclerotium against AFB1-induced hepato- and geno-toxicity --- p.126
Chapter 3.3.2.1 --- Dose-response of AFB1 on rat model --- p.126
Chapter 3.3.2.2 --- Preventive test of Ptr sclerotium against AFB1-induced geno- and hepatotoxicity --- p.134
Chapter 3.3.2.3 --- CHL versus 30% Ptr sclerotial diet --- p.137
Chapter 3.3.3 --- A comparison of the hepatotoxicity of CC14 and AFB1 --- p.142
Chapter 3.4 --- Summary --- p.147
Chapter Chapter 4 --- Conclusions and future work --- p.148
References --- p.151
Related publication --- p.175

At present, there is no cure for bone metastasis. The current goals in patient care are to palliate pain, prevent pathological bone fracture and increase the strength and function of bone, so as to extend the life expectancy and maintain a good quality of life. Bisphosphonate treatment is the currently standard therapy of bone metastasis and is commonly used by physicians; it alleviates the tumour-induced hypercalcemia in 90% of patients and reduces the metastatic bone pain in 50% of patients. Moreover, it also prevents the pathological fracture of the affected bones. However, while effective, bisphosphonate injections are very costly, though its oral formulation is less expensive it is also less efficacious, and causes gastrointestinal discomfort. Furthermore, prolonged use of bisphosphonate treatment may lead to certain adverse effects, including hypocalcemia. These factors will prohibit the longterm use of such medication as it can negatively affect the treatment outcome.
Based on enormous medical potentials illustrated by the aforementioned findings, BBYNG deserves wider clinical application, large-scale clinical study on its preventive effect against bone metastasis and detailed investigation of its mode(s) of action in the body.
Based on the above-described understanding of Chinese medicine and bone metastasis, supplementing the kidney and strengthening bone could be the basic principle for the treatment of bone metastasis using Chinese medicine. In view of this theory, and in addition to the clinical observation and a thorough search of the available literature, we selected relevant kidney-tonifying Chinese herbs, namely (Fructus Ligustri Lucidi), (Rhizoma Drynariae), (Herba Epimedii), (Psoralea Corylifolia) and wide-spectrum anticancer herbs (Herba Hedyotidis Diffusae) for the preparation of a combined formula--BBYNG.
Chinese medicine has long been used to treat cancers. Its advantages reside in its holistic properties, which bring palliative, corrective and convalescing functions against damage caused by radiotherapy, chemotherapy and surgery. These features position Chinese medicines as the adjuvant to orthodox cancer treatment. During the late stage of tumour development, when standard therapy is no longer effective, Chinese medicine plays a critical role as an integrated therapy. Searching for a safe, inexpensive and effective Chinese medicine preparation suitable for prolonged use as adjunct therapy in late cancer cases is of paramount importance.
Clinical results. Both Chinese medicine and Western medicine treated patients showed no significant change in their blood parameters or liver and kidney examinations before and after drug administration; Male subjects on BBYNG, their bone mass density remained stable after 6 months treatment and the subjects on OSTAC showed slightly decreased In females, subjects on BBYNG remained stable, but subjects on OSTAC slightly increased.
Clinical study. The study was designed as a randomized, parallel-group comparison between BBYNG formula and Bisphosphonate. The patients who meet the inclusion/exclusion criteria were randomly assigned to receive either BBYNG granules, which was prepared by a GMP manufacturer, or Clodronic acid. The treatment period was 6 months (24 weeks). For both groups, various clinical parameters such as body functions, blood examinations, bone density (BMD) assessment, X-ray examinations, pain intensity and quality of life were evaluated and compared.
Conclusions. (1) As an adjuvant to patients with bone metastases, BBYNG is effective in relieving the metastatic bone pain, improving the quality of life. (2) In the animal model, BBYNG reduced the metastatic bone damage, prolonged the survival and enhanced the T lymphocyte immunity in the tumour-bearing mice. (3) In vitro study on the breast and lung cancer cell lines showed that BYYNG could induce apoptosis and prevent tumour cell invasion. It suggests that BYYNG may restrict tumour growth and development, thus reducing the occurrence of bone metastasis.
In accordance with Chinese medicine, bone metastasis can be categorized into "bone tumour" "bone erosion" "bone wilting" "bone necrosis" and "bone impediment". The main cause of bone metastasis is twofold: cancer toxicity, and in Chinese medicine theory, the kidney governs the bone marrow, if the kidney is not functioning in balance, then the bone will become weak. Cancer toxicity is the "pathogenic cause" to skeletal metastases, while kidney weakness decreases the body defence against the cancer. A vicious cycle ensues when cancer and kidney deficiency and bone weakness occurs simultaneously coincidently and worsens the conditions.
In vitro study on tumour cell lines. The anticancer effects of different concentrations of BBYNG formula and various single components against human breast cancer and lung cancer cell lines were evaluated by cell viability test (MTT assay), cell apoptosis test and invasion suppression test.
In vitro study results. BBYNG and the aqueous extracts of its component herbs at very low drug concentrations stimulated the growth of three tumour cell lines tested. When the concentrations were slightly increased, they showed an inhibitory effect on cancer cell proliferation. As the drug concentrations further increased, the extracts showed cytotoxic effects on these tumor cells. At the noncytotoxic dose, the extracts could trigger apoptosis and enhance the caspase-3 activity in all three tumour cell lines. In addition, at this "non toxic" concentration, the extracts markedly inhibited the in vitro invasive property of the 4T1 breast cancer cell lines in our Matrigel invasion model. Thus these in vitro results suggested that BBYNG possess anticancer, invasion-inhibitory and anti-metastatic activities.
In vivo animal study results. (Tumour growth was slower in the BBYNG treatment group when compared to the OSTAC and control groups, but this was not significantly difference) BBYNG significantly delayed tumour growth in tumour bearing mice, but it did not minimize the tumour size markedly. Moreover, BBNYG did minimize the mobility restriction caused by tumours, reduce the damage to bones, prolong the survival time and enhanced the T lymphocyte immunity.
In vivo animal study. A well-established animal model for breast cancer was used to evaluate and compare the pharmacological effects of BBYNG formula and Clodronic acid, as shown by different indicators such as tumour progression, animal's mobility, survival time, bone metastasis-induced fracture intensity and the immunological status of the tumour-bearing mice.
Malignant tumour is characterized by early metastasis. Among them 37 to 80 (depending on which type of cancer) patients show tendency of bone metastasis. Bone metastasis is usually accompanied by various complications, such as severe pain, pathological bone fracture, hypercalcemia, and bone marrow suppression, which can substantially affect the quality of life of the patients. Thus, the prevention and treatment of bone metastasis in cancer is an issue worth pursuing.
Malignant tumours leading to high mortality and morbidity are a serious threat to human health. It is the leading cause of death in China. In Hong Kong, there are over 20 thousand new cancer cases and more than 1100 people die due to cancers every year.
Study objectives. To elucidate the efficacy and some pharmacological aspects of BBYNG in regard to the treatment of bone metastasis through clinical observation and different laboratory experiments. This study would be of significant reference value to the disease-oriented drug formulation and application, mechanistic study and research methodology of the treatment of bone metastasis using Chinese medicines.
The clinical and laboratory experimental results are summarized as below:
The research study is composed of three parts, the clinical study, in vivo animal study and in vitro study on tumour cell lines. The research methods used are as follows:
Those on BBYNG treatment showed more a stable and satisfactory quality of life than those in the Western medicine-treated group. For the Clodronic acid treatment group, patients generally showed worsened symptoms and quality of life deteriorated. The ECOG index of the BBYNG group was statistically better than that of the Clodronic acid group. Within the 72-week clinical observational period, the mortality of Clodronic acid group is significantly higher that of the BBYNG group. The effects of BBYNG group as presented in relieving the pain-induced influence on patients' emotion, interpersonal relationship and entertainment was more pronounced than that in the Clodronic acid group.
Wu Ka.
論文(哲學博士)--香港中文大學, 2006.
參考文獻(p. 299-324).
Adviser: Leung Ping Chung.
Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1570.
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in Chinese and English.
School code: 1307.
Lun wen (zhe xue bo shi)--Xianggang Zhong wen da xue, 2006.
Can kao wen xian (p. 299-324).
Wu Ka.

Ren-Wang Jiang.
"July 2002."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references.
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

Yuen Yee Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 108-125).
Abstracts in English and Chinese.
TITLE PAGE --- p.1
ACKNOWLEGDEMENTS --- p.2
ABSTRACT --- p.3
摘要 --- p.5
table of contents --- p.7
list of figures and tables --- p.13
CHAPTER 1 GENERAL INTRODUCTION --- p.16
Chapter 1.1 --- role of PHYTOESTROGENS in soy and red WINE the PREVENTION OF CARDIOVASCULAR DISEASES (CVD) --- p.17
Chapter 1.1.1 --- INTRoduction and Classification of Phytoestrogens --- p.17
Chapter 1.1.2 --- estrogenic1ty of phytoestrogens and theIr abundancesin Plasma --- p.18
Chapter 1.1.3 --- phytoestrogens as one of the active components In cvd Protection --- p.21
Chapter 1.1.4 --- effects of Phytoestrogens on LDL Receptor and Apolipoprotein A-1 --- p.22
Chapter 1.2 --- role of estrogen receptors (ers) in gene regulation --- p.24
Chapter 1.2.1 --- "structure, Classification and tissue distribution of ERS" --- p.24
Chapter 1.2.2 --- ligands for ERS --- p.25
Chapter 1.2.3 --- mechaniSMS OF LIgands-ERS complex in GENE regulation --- p.27
Chapter 1.2.4 --- ligand-independent ER activation --- p.28
Chapter 1.3 --- aims and scopes of investigation --- p.29
Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.30
Chapter 2.1 --- chemicals and materials --- p.30
Chapter 2.1.1 --- Chemicals --- p.30
Chapter 2.1.2 --- Plasmids --- p.30
Chapter 2.2 --- mammalian cell culture maintainence --- p.30
Chapter 2.2.1 --- Maintenance of Cells --- p.31
Chapter 2.2.2 --- Preparation of Cell Stock --- p.31
Chapter 2.2.3 --- Cell Recovery from Liquid Nitrogen Stock --- p.31
Chapter 2.3 --- manipulation of dna --- p.31
Chapter 2.3.1 --- isolation of HEPG2 cells genonmic DNA --- p.31
Chapter 2.3.2 --- separation and purification of dna from agarose gel --- p.31
Chapter 2.3.3 --- Restriction digestionof DNA --- p.32
Chapter 2.3.4 --- Ligation of DNA Fragments --- p.32
Chapter 2.3.5 --- Transformation of --- p.32
Chapter 2.3.6 --- Small Scale Plasmids Purification from DH5a --- p.32
Chapter 2.4 --- construction of expression and reporter plasmids --- p.33
Chapter 2.4.1 --- Construction of Estrogen Receptorα (Erα) Expression Vectors --- p.33
Chapter 2.4.2 --- construction of reporter vectors of LDLR promoter and the Respective Mutants --- p.33
Chapter 2.4.3 --- Construction of Reporter Vectors of APOAI Promoter and the Respective Mutants --- p.33
Chapter 2.5 --- determination of promoter transcrtiption activities --- p.34
Chapter 2.5.1 --- Transient Transfection of Cell with ERa Expression Vector and Promoter Reporter using Lipofectamine PLUS Reagent --- p.34
Chapter 2.5.2 --- Dual Luciferase Assay --- p.34
Chapter 2.6 --- semi-quantitative and quantitative rt-pcr assay --- p.34
Chapter 2.6.1 --- Transient transfection of Cell with ERa Expression Vector Using Lipofectamine PLUS Reagent --- p.34
Chapter 2.6.2 --- "Isolation of RNA using TRIzol® Reagent (Life Technology, USA)" --- p.35
Chapter 2.6.3 --- Quantitation of RNA --- p.35
Chapter 2.6.4 --- First Strand cDNA Synthesis --- p.35
Chapter 2.6.5 --- Sem卜Quantitative PCR Reactions --- p.35
Chapter 2.6.6 --- Quantitative PCR Reactions --- p.36
Chapter 2.7 --- western blotting analysis --- p.36
Chapter 2.8 --- statistical methods --- p.36
Chapter CHAPTER 3 --- REGULATION BY PHYSIOLOGICAL LEVEL OF 17B-ESTRADIOL ON APOLIPOPROTEIN A-I AND LOW-DENSITY- LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.37
Chapter 3.1 --- introduction --- p.37
Chapter 3.2 --- results --- p.39
Chapter 3.2.1 --- Determination of transient transfection functionality of estrogen receptors in hepg2 cells --- p.39
Chapter 3.2.2 --- Effect of 17β-Estradiolon LDLR promoter transcription activity --- p.39
Chapter 3.2.3 --- Effect of 17β-Estradiol on apoai promoter transcription activity --- p.40
Chapter 3.2 --- discussion --- p.47
Chapter CHAPTER 4 --- SOY ISOFLAVONES AND RESVERATROL DISPLAY DIFFERENT MECHANISM IN THE UP-REGULATION OF LOVV-DENSITY-LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.49
Chapter 4.1 --- introduction --- p.49
Chapter 4.2 --- results --- p.54
Chapter 4.2.1 --- Association of ERα and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54
Chapter 4.2.2 --- Association of ERβ and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54
Chapter 4.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol induced LDLR promoter transcription" --- p.55
Chapter 4.2.4 --- Identification of promoter regions responsible for induction of LDLR transcription by isoflavones in the presence OF ERα --- p.55
Chapter 4.2.5 --- Identification of promoter regions responsible for induction of LDLR TRANSCRIPTION BY resveratrol IN THE ABSENCE OF ERα --- p.56
Chapter 4.3 --- DISCUSSION --- p.75
Chapter CHAPTER 5 --- SOY ISOFLAVONES AND RESVERATROL UP-REGULATE APOLIPOPROTEIN A-I SIMILAR TO 17B-ESTRADIOL IN HEPG2 CELLS --- p.80
Chapter 5.1 --- INTRODUCTION --- p.80
Chapter 5.2 --- RESULTS --- p.84
Chapter 5.2.1 --- Association of ERα phytoestrogens on APCAI gene expression --- p.84
Chapter 5.2.2 --- Association of ERβ and isoflavones or resveratrol on APOAI promoter transcription activity --- p.85
Chapter 5.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol in APOAI promoter transcription in the presence of ERα" --- p.85
Chapter 5.2.4 --- Identification of promoter regions responsible for induction of APOAI transcription by isoflavones and resveratrol in the presence of ERα --- p.85
Chapter 5.3 --- DISCUSSION --- p.100
Chapter CHAPTER 6 --- GENERAL DISCUSSION --- p.103
Chapter CHAPTER 7 --- SUMMARY --- p.106
BIBLIOGRAPHY --- p.108
APPENDIX 1 ABBREVIATIONS --- p.126
APPENDIX 2 MATERIALS AND METHODS --- p.129
APPENDIX 3 PRIMER LISTS --- p.145
APPENDIX 4 REAGENTS AND BUFFERS --- p.147

Wong Chun-kwan.
Thesis submitted in: December 1999.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2000.
Includes bibliographical references (leaves 127-137).
Abstracts in English and Chinese.
Abstract --- p.i
Acknowledgments --- p.viii
Tables of Contents --- p.ix
List of Figures --- p.xv
List of Tables --- p.xxvi
Chapter Chapter 1: --- INTRODUCTION --- p.1
Chapter Chapter 2: --- LITERATURE REVIEW --- p.8
Chapter 2.1 --- Toxicology --- p.8
Chapter 2.1.1 --- Acute toxicity test --- p.8
Chapter 2.1.2 --- Biochemical Analysis --- p.9
Chapter 2.1.3 --- Organ weights --- p.10
Chapter 2.2 --- Histology --- p.11
Chapter 2.2.1 --- Light Microscope --- p.11
Chapter 2.2.2 --- Electron Microscopy --- p.11
Chapter 2.3 --- Tissue injury --- p.12
Chapter 2.3.1 --- Free-radical mechanisms --- p.12
Chapter 2.3.2 --- Lipid peroxidation --- p.13
Chapter 2.4 --- Carbon tetrachloride (CC14) --- p.14
Chapter 2.4.1 --- Mechanisms of carbon tetrachloride toxicity --- p.15
Chapter 2.5 --- Trichloroethylene (TCE) --- p.18
Chapter 2.5.1 --- Mechanisms of trichloroethylene toxicity --- p.21
Chapter 2.6 --- Dimethyl sulfoxide (DMSO) --- p.25
Chapter 2.7 --- N-acetylcysteine (NAC) --- p.27
Chapter Chapter 3: --- MATERIALS AND METHODS --- p.28
Chapter 3.1 --- Materials --- p.28
Chapter 3.2 --- Methods --- p.31
Chapter 3.2.1 --- Acute hepatotoxicity test on aqueous seaweed extracts --- p.31
Chapter 3.2.1.1 --- Preparation of aqueous extracts of seaweed --- p.31
Chapter 3.2.1.2 --- Experimental protocol --- p.31
Chapter 3.2.1.3 --- Biochemical assays --- p.32
Chapter 3.2.1.4 --- Organ weights --- p.36
Chapter 3.2.1.5 --- Histopathological examination --- p.36
Chapter 3.2.1.6 --- Statistical analysis --- p.36
Chapter 3.2.2 --- Curative and preventive tests of seaweed aqueous extracts against the CCl4-induced hepatotoxicity --- p.37
Chapter 3.2.2.1 --- Preparation of aqueous extracts of seaweed --- p.37
Chapter 3.2.2.2 --- Experimental protocol --- p.37
Chapter 3.2.2.3 --- Biochemical assays --- p.39
Chapter 3.2.2.4 --- Organ weights --- p.39
Chapter 3.2.2.5 --- Histopathological examination --- p.40
Chapter 3.2.2.6 --- Statistical analysis --- p.41
Chapter 3.2.3 --- Acute hepatotoxicity test of TCE in rats by oral and intraperitoneal routes --- p.42
Chapter 3.2.3.1 --- Experimental protocol --- p.42
Chapter 3.2.3.2 --- Biochemical assays --- p.43
Chapter 3.2.3.3 --- Organ weights --- p.43
Chapter 3.2.3.4 --- Histopathological examination --- p.44
Chapter 3.2.3.5 --- Statistical analysis --- p.44
Chapter 3.2.4 --- Curative and preventive tests of seaweed aqueous extracts against the TCE effective dose-induced toxicity --- p.44
Chapter 3.2.4.1 --- Preparation of aqueous extracts of seaweed --- p.44
Chapter 3.2.4.2 --- Experimental protocol --- p.45
Chapter 3.2.4.3 --- Biochemical assays --- p.46
Chapter 3.2.4.4 --- Organ weights --- p.46
Chapter 3.2.4.5 --- Histopathological examination --- p.46
Chapter 3.2.5 --- Antidotal effects of dimethyl sulfoxide (DMSO) and N-acetylcysteine (NAC) against CC14- and TCE- induced poisoning in rats --- p.47
Chapter 3.2.5.1 --- Experimental protocol --- p.47
Chapter 3.2.5.2 --- Biochemical assays --- p.48
Chapter 3.2.5.3 --- Organ weights --- p.48
Chapter 3.2.5.4 --- Histopathological examination --- p.49
Chapter 3.2.6 --- Hepatoprotective effect of seaweeds' methanol extract against CC14- and TCE-induced poisoning in rats --- p.49
Chapter 3.2.6.1 --- Preparation of methanol extracts of seaweed --- p.49
Chapter 3.2.6.2 --- Experimental protocol --- p.50
Chapter 3.2.6.3 --- Biochemical assays --- p.52
Chapter 3.2.6.4 --- Organ weights --- p.52
Chapter 3.2.6.5 --- Histopathological examination --- p.53
Chapter Chapter 4 --- RESULTS --- p.54
Chapter 4.1 --- Acute hepatotoxicity test on aqueous seaweed extracts --- p.54
Chapter 4.1.1 --- The biochemical assays of the serum transaminase activity --- p.54
Chapter 4.1.2 --- The organ weight (Aqueous seaweed crude extracts) --- p.56
Chapter 4.2 --- Curative and preventive tests of seaweed aqueous extracts against the CCl4-induced hepatotoxicity --- p.58
Chapter 4.2.1 --- The biochemical assays of the serum transaminase activity (Curative) --- p.58
Chapter 4.2.2 --- The organ weight (Curative) --- p.60
Chapter 4.2.3 --- The biochemical assays of the serum transaminase activity (Preventive) --- p.62
Chapter 4.2.4 --- The organ weight (Preventive) --- p.64
Chapter 4.3 --- Acute hepatotoxicity test of TCE in rats by oral and intraperitoneal routes --- p.66
Chapter 4.3.1 --- Oral route --- p.66
Chapter 4.3.1.1 --- One-time oral route --- p.66
Chapter 4.3.1.2 --- Two-time oral route --- p.66
Chapter 4.3.2 --- Intraperitoneal route --- p.66
Chapter 4.3.3 --- Time course of the effective dose of 20% TCE in i.p. route --- p.67
Chapter 4.4 --- Curative and preventive tests of seaweed aqueous extracts against the TCE effective dose-induced toxicity --- p.12
Chapter 4.4.1 --- The biochemical assays of the serum transaminase activity (Curative) --- p.72
Chapter 4.4.2 --- The organ weight (Curative) --- p.74
Chapter 4.4.3 --- The biochemical assays of the serum transaminase activity (Preventive) --- p.76
Chapter 4.4.4 --- The organ weight (Preventive) --- p.78
Chapter 4.5 --- Antidotal effects of dimethyl sulfoxide (DMSO) and N-acetylcysteine (NAC) against CC14- and TCE-induced poisoning in rats --- p.80
Chapter 4.5.1 --- The biochemical assays of the serum transaminase activity (Curative) --- p.80
Chapter 4.5.2 --- The organ weight (Curative) --- p.82
Chapter 4.5.3 --- The biochemical assays of the serum transaminase activity (Preventive) --- p.84
Chapter 4.5.4 --- The organ weight (Preventive) --- p.86
Chapter 4.6 --- Hepatoprotective effect of methanol extract of seaweed against CC14- and TCE-induced poisoning in rats --- p.88
Chapter 4.6.1 --- The biochemical assays of the serum transaminase activity (Curative) --- p.88
Chapter 4.6.2 --- The organ weight (Curative) --- p.89
Chapter 4.7 --- Histopathological examinations --- p.90
Chapter 4.7.1 --- Acute hepatotoxicity test on aqueous seaweed extracts --- p.91
Chapter 4.7.2 --- Curative and preventive tests of seaweed aqueous extracts against the CC14-induced hepatotoxicity --- p.92
Chapter 4.7.3 --- Acute hepatotoxicity test of TCE in rats by oral and intraperitoneal routes --- p.99
Chapter 4.7.4 --- Curative and preventive tests of seaweed aqueous extracts against the TCE effective dose-induced toxicity --- p.100
Chapter 4.7.5 --- Antidotal effects of dimethyl sulfoxide (DMSO) and N-acetylcysteine (NAC) against CC14- and TCE-induced poisoning in rats --- p.100
Chapter 4.7.6 --- Hepatoprotective effect of methanol extract of seaweed against CC14- and TCE-induced poisoning in rats --- p.102
Chapter Chapter 5 --- DISCUSSION --- p.106
Chapter Chapter 6 --- CONCLUSION --- p.124
REFERENCES --- p.127
APPENDIX --- p.138

Lee, Ching Ching.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 90-103).
Abstracts in English and Chinese.
Acknowledgments --- p.i
Abstract --- p.ii
論文摘要 --- p.iv
Table of Contents --- p.vi
List of Tables --- p.x
List of Figures --- p.xi
Chapter Chapter 1 --- Introduction
Chapter 1.1. --- Quinolone Antimicrobial Agents --- p.1
Chapter 1.1.1. --- Development --- p.1
Chapter 1.1.2. --- Mode of action --- p.3
Chapter 1.1.3. --- Mechanisms of resistance to quinolones --- p.4
Chapter 1.1.3.1. --- Target genes mutations --- p.4
Chapter 1.1.3.2. --- Decreased intracellular quinolone accumulation --- p.5
Chapter 1.1.3.3. --- Plasmid-mediated quinolone resistance --- p.6
Chapter 1.2. --- Plasmid-mediated Quinolone Resistance Genes (qnr) --- p.8
Chapter 1.2.1. --- Discovery of qnrA genes --- p.8
Chapter 1.2.2. --- Discovery of qnrS genes --- p.9
Chapter 1.2.3. --- Discovery of qnrB genes --- p.10
Chapter 1.2.4. --- Discovery of qnrC genes --- p.11
Chapter 1.2.5. --- Discovery of qnrD genes --- p.12
Chapter 1.2.6. --- Origins of qnr genes --- p.12
Chapter 1.2.7. --- Qnr proteins and mode of action --- p.14
Chapter 1.2.8. --- Epidemiology and quinolones resistance activity of qnr genes --- p.16
Chapter 1.2.9. --- Epidemiology of fluoroquinolone-resistant Enterobacteriaceae --- p.17
Chapter 1.2.10 --- Multidrug-resistant in extended-spectrum-B-lactamase- and AmpC-producing Enterobacteriaceae --- p.19
Chapter 1.3. --- Background of Study --- p.20
Chapter 1.4. --- Objectives of Study --- p.21
Chapter Chapter 2 --- Materials & Methods
Chapter 2.1. --- Study Design --- p.22
Chapter 2.2. --- Antimicrobial Susceptibility Testing --- p.24
Chapter 2.2.1 --- Bacterial isolates --- p.24
Chapter 2.2.2. --- Screening for ESBL and AmpC production by disk diffusion test --- p.24
Chapter 2.2.3. --- Determination of minimal inhibitory concentrations (MICs) --- p.25
Chapter 2.3. --- "Detection of qnrA, qnrB and qnrS Genes by Multiplex PCR" --- p.27
Chapter 2.3.1. --- Total DNA preparation --- p.27
Chapter 2.3.2. --- "Multiplex PCR assay for qnrA, qnrB and qnrS genes detection" --- p.27
Chapter 2.3.3. --- Agarose gel electrophoresis --- p.29
Chapter 2.4. --- "Detection of TEM-, SHV-, CTX- and PMAmpC Type B-Lactamase Genes by PCR" --- p.30
Chapter 2.5. --- PCR Assays for Further Genotypic Characterization Purpose --- p.32
Chapter 2.5.1. --- PCR assay to amplify qnrB genes --- p.32
Chapter 2.5.2. --- PCR assay to amplify qnrS genes --- p.33
Chapter 2.5.3. --- "PCR assays for genotypic characterizations of the co-existed blaTEM, blaSHV, blaCTX-M and PMAmpC genes of all qnr-positive isolates" --- p.33
Chapter 2.5.3.1. --- Genotypic characterizations of the co-existed bla-TEM and genes --- p.33
Chapter 2.5.3.2. --- PCR assays to amplify the co-existed blaCTX_M genes --- p.33
Chapter 2.5.3.3. --- PCR assay to amplify the co-existed PMAmpC genes --- p.34
Chapter 2.5.4. --- Sequencing reaction --- p.36
Chapter 2.5.4.1. --- Purification of PCR product and sequence determination --- p.36
Chapter 2.5.4.2. --- Sequence analysis --- p.37
Chapter 2.6. --- Collection of Clinical Data --- p.38
Chapter 2.6.1. --- Demographics and clinical data --- p.38
Chapter 2.6.2. --- Definitions --- p.38
Chapter 2.6.3. --- Data analysis --- p.40
Chapter Chapter 3 --- Results
Chapter 3.1. --- Bacterial Isolates --- p.41
Chapter 3.2. --- "Demographics, Medical History, Clinical Features and Clinical Outcomes of Patients" --- p.42
Chapter 3.3. --- Antimicrobial Susceptibility Testing --- p.44
Chapter 3.4. --- Detection of qnr Genes --- p.48
Chapter 3.4.1. --- "Detection of qnrA, qnrB and qnrS genes by multiplex PCR" --- p.48
Chapter 3.5. --- Detection of ESBLs --- p.49
Chapter 3.5.1. --- Detection of TEM- and SHV-type ESBLs --- p.49
Chapter 3.5.2. --- Detection of CTX-M- type ESBLs --- p.51
Chapter 3.6. --- Detection of PMAmpC Genes --- p.52
Chapter 3.6.1. --- Detection of PMAmpC genes --- p.52
Chapter 3.7. --- "The Distribution of qnr and bla Genes for TEM, SHV, CTX-M and PMAmpC" --- p.53
Chapter 3.8. --- The Characteristics of qnr Isolates --- p.54
Chapter 3.8.1. --- Genotypes of qnrB and qnrS --- p.54
Chapter 3.8.2. --- Antimicrobial susceptibility of qnr isolates --- p.58
Chapter 3.9. --- "The Associations of qnr Genes with Other Bacterial Resistance Genotypes, and the Clinical Characteristics and Outcomes of Patients" --- p.62
Chapter 3.9.1. --- "Univariate analysis of the associations of qnr genes with other bacterial resistance genotypes, and the clinical characteristics and outcomes of patients" --- p.62
Chapter 3.9.2. --- "Multivariate analysis of the associations of qnr genes with other bacterial resistance genotypes, and the clinical characteristics and outcomes of patients" --- p.65
Chapter 3.9.2.1. --- "Multivariate analysis of the associations of qnr genes with other bacterial resistance genotypes, and the clinical characteristics of patients" --- p.65
Chapter 3.9.2.2. --- "Multivariate analysis of the associations of mortality with qnr genes, bacterial resistance genotypes and other clinical characteristics of patients" --- p.66
Chapter Chapter 4 --- Discussion
Chapter 4.1. --- Prevalences and Susceptibility of ESBL and PMAmpC in Bacteraemic Enterobacteriaceae Isolates --- p.67
Chapter 4.2. --- Epidemiology of Plasmid-mediated Quinolone Resistance (qnr) Genes --- p.69
Chapter 4.3. --- Genotypes of qnr-positive Isolates --- p.72
Chapter 4.4. --- Antimicrobial Susceptibility of qnr-positive Isolates --- p.75
Chapter 4.5. --- "The Associations of qnr Genes with Other Bacterial Resistance Genotypes, and the Clinical Characteristics of Patients" --- p.79
Chapter 4.6. --- "The Associations of Mortality with qnr Genes, Bacterial Resistance Genotypes and Other Clinical Characteristics of Patients" --- p.80
Chapter 4.7. --- Clinical Importance and Clinical Implications of qnr Genes --- p.82
Chapter 4.8. --- Limitations of the Current Study --- p.85
Chapter 4.9. --- Future Studies --- p.87
Chapter 4.10. --- Conclusions --- p.89
References --- p.90

Ho Toi Sze Joyce.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 65-74).
Abstracts in English and Chinese.
Acknowledgement --- p.ii
Contents --- p.iii
Abstract --- p.viii
List of Abbreviations --- p.xvii
List of Tables --- p.xix
List of Figures --- p.xx
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- The role of Non-steroidal anti-inflammatory drugs (NSAIDs) --- p.1
Chapter 1.2 --- NSAID-induced gastrointestinal (GI) toxicity --- p.1
Chapter 1.2.1 --- Pathogenesis of NSAID-induced GI toxicity --- p.2
Chapter 1.2.2 --- GI symptoms --- p.4
Chapter 1.2.3 --- GI ulcers --- p.4
Chapter 1.2.4 --- GI complications --- p.5
Chapter 1.2.5 --- Risk factor for GI complications --- p.6
Chapter 1.2.6 --- Ulcerogenicity of different NSAIDs in upper GI events --- p.6
Chapter 1.3 --- Prevention of NSAID-induced GI toxicity --- p.7
Chapter 1.3.1 --- H2-receptor antagonists --- p.8
Chapter 1.3.2 --- Misoprostol --- p.8
Chapter 1.3.3 --- Proton Pump Inhibitor (PPI) --- p.9
Chapter 1.3.4 --- Selective COX-2 Inhibitors --- p.10
Chapter 1.3.4.1 --- GI safety of selective COX-2 inhibitors --- p.11
Chapter 1.3.4.1.1 --- Gastrointestinal outcomes research of rofecoxib --- p.13
Chapter 1.3.4.1.2 --- Celecoxib Long term Arthritis Safety Study --- p.14
Chapter 1.3.4.2 --- Cardiovascular toxicity of NSAIDs --- p.15
Chapter 1.3.4.2.1 --- Cardiovascular toxicity of non-selective NSAIDs --- p.15
Chapter 1.3.4.2.2 --- Cardiovascular toxicity of selective COX-2 inhibitors --- p.16
Chapter 1.4 --- Guidelines on the management of osteoarthritis (OA) and rheumatoid arthritis (RA) --- p.21
Chapter 1.4.1 --- American College of Rheumatology (ACR) Subcommittee --- p.22
Chapter 1.4.2 --- National Institute for Clinical Excellence (NICE) --- p.23
Chapter 1.4.3 --- Hong Kong Hospital Authority (HA) --- p.23
Chapter 1.5 --- Cost of illness of upper GI events in the setting of an emergency room of a regional hospital in Hong Kong and cost analysis of selective COX-2 inhibitor with non-selective NSAID plus gastroprotective agent --- p.24
Chapter 1.6 --- Objectives --- p.25
Chapter Chapter 2 --- Cost of illness of upper GI events in the setting of an emergency room of a regional hospital in Hong Kong --- p.26
Chapter 2.1 --- Methods --- p.28
Chapter 2.1.1 --- Study site --- p.28
Chapter 2.1.2 --- Cohort participants --- p.28
Chapter 2.1.3 --- Resource data collection --- p.29
Chapter 2.1.4 --- Cost data --- p.30
Chapter 2.1.5 --- Statistical Methods --- p.31
Chapter 2.1.6 --- Study perspective --- p.31
Chapter 2.2 --- Results --- p.31
Chapter 2.2.1 --- Demographic data --- p.31
Chapter 2.2.2 --- Total direct medical cost of upper GI complaints in UCH --- p.33
Chapter 2.3 --- Discussion --- p.35
Chapter 2.3.1 --- Total direct medical cost of upper GI events --- p.35
Chapter 2.3.2 --- Cost of upper GI events associated with NSAID usage --- p.38
Chapter 2.3.3 --- Low dose aspirin on NSAID-induced GI toxicity --- p.38
Chapter 2.3.4 --- Limitation --- p.39
Chapter 2.3.5 --- Future study --- p.41
Chapter 2.4 --- Conclusion --- p.41
Chapter Chapter 3 --- Cost analysis of selective COX-2 inhibitor versus non-selective NSAID with gastroprotective agent --- p.43
Chapter 3.1 --- Methods --- p.46
Chapter 3.1.1 --- Local randomized clinical trial --- p.46
Chapter 3.1.1.1 --- Study population --- p.46
Chapter 3.1.1.2 --- Cost data --- p.47
Chapter 3.1.1.3 --- Statistical Methods --- p.48
Chapter 3.1.1.4 --- Sensitivity analysis --- p.49
Chapter 3.1.2 --- Large randomized clinical trial --- p.49
Chapter 3.1.2.1 --- Study population --- p.49
Chapter 3.1.2.2 --- Cost data --- p.50
Chapter 3.2 --- Results --- p.50
Chapter 3.2.1 --- Local randomized clinical trial --- p.51
Chapter 3.2.1.1 --- Demographic data --- p.51
Chapter 3.2.1.2 --- Cost analysis --- p.52
Chapter 3.2.1.3 --- Sensitivity analysis --- p.53
Chapter 3.2.2 --- Large randomized clinical trial --- p.54
Chapter 3.2.2.1 --- Demographic data --- p.54
Chapter 3.2.2.2 --- Cost analysis --- p.55
Chapter 3.3 --- Discussion --- p.55
Chapter 3.3.1 --- Cost analysis --- p.55
Chapter 3.3.2 --- Sensitivity analysis --- p.59
Chapter 3.3.3 --- Low dose aspirin on NSAID-induced GI toxicity --- p.59
Chapter 3.3.4 --- Limitation --- p.60
Chapter 3.4 --- Future study --- p.62
Chapter 3.5 --- Conclusion --- p.62
Chapter Chapter 4 --- Conclusion --- p.63
Chapter Chapter 5 --- Reference --- p.65
Appendix Data collection form --- p.75

BACKGROUND: Although guidelines indicate when patients are eligible for antihypertensives and statins, little is known about whether general practitioners (GPs) follow this guidance. OBJECTIVE: To determine the factors influencing GPs decisions to prescribe cardiovascular prevention drugs. DESIGN OF STUDY: Secondary analysis of data collected on patients whose cardiovascular risk factors were measured as part of a controlled study comparing nurse-led risk assessment (four practices) with GP-led risk assessment (two practices). SETTING: Six general practices in the West Midlands, England. PATIENTS: Five hundred patients: 297 assessed by the project nurse, 203 assessed by their GP. MEASUREMENTS: Cardiovascular risk factor data and whether statins or antihypertensives were prescribed. Multivariable logistic regression models investigated the relationship between prescription of preventive treatments and cardiovascular risk factors. RESULTS: Among patients assessed by their GP, statin prescribing was significantly associated only with a total cholesterol concentration >/= 7 mmol/L and antihypertensive prescribing only with blood pressure >/= 160/100 mm Hg. Patients prescribed an antihypertensive by their GP were five times more likely to be prescribed a statin. Among patients assessed by the project nurse, statin prescribing was significantly associated with age, sex, and all major cardiovascular risk factors. Antihypertensive prescribing was associated with blood pressures >/= 140/90 mm Hg and with 10-year cardiovascular risk. LIMITATIONS: Generalizability is limited, as this is a small analysis in the context of a specific cardiovascular prevention program. CONCLUSIONS: GP prescribing of preventive treatments appears to be largely determined by elevation of a single risk factor. When patients were assessed by the project nurse, prescribing was much more consistent with established guidelines.

Woo Yiu Ho Anthony.
"August 2004."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (p. 176-198).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

Indiana University-Purdue University Indianapolis (IUPUI)
Human induced pluripotent stem (iPS) cells have the unique ability to differentiate into 200 or so somatic cell types that make up the adult human being. The use of human iPS cells to study development and disease is a highly exciting and interdependent field that holds great promise in understanding and elucidating mechanisms behind cellular differentiation with future applications in drug screening and cell replacement studies for complex and currently incurable cellular degenerative disorders. The recent advent of iPS cell technology allows for the generation of patient-specific cell lines that enable us to model the progression of a disease phenotype in a human in vitro model. Differentiation of iPS cells toward the affected cell type provides an unlimited source of diseased cells for examination, and to further study the developmental progression of the disease in vitro, also called the “disease-in-a-dish” model. In this study, efforts were undertaken to recapitulate the differentiation of distinct retinal cell affected in two highly prevalent retinal diseases, Usher syndrome and glaucoma. Using a line of Type III Usher Syndrome patient derived iPS cells efforts were undertaken to develop such an approach as an effective in vitro model for studies of Usher Syndrome, the most commonly inherited disorder affecting both vision and hearing. Using existing lines of iPS cells, studies were also aimed at differentiation and characterization of the more complex retinal cell types, retinal ganglion cells (RGCs) and astrocytes, the cell types affected in glaucoma, a severe neurodegenerative disease of the retina leading to eventual irreversible blindness. Using a previously described protocol, the iPS cells were directed to differentiate toward a retinal fate through a step-wise process that proceeds through all of the major stages of neuroretinal development. The differentiation process was monitored for a period of 70 days for the differentiation of retinal cell types and 150 days for astrocyte development. The different stages of differentiation and the individually derived somatic cell types were characterized by the expression of developmentally associated transcription factors specific to each cell type. Further approaches were undertaken to characterize the morphological differences between RGCs and other neuroretinal cell types derived in the process. The results of this study successfully demonstrated that Usher syndrome patient derived iPS cells differentiated to the affected photoreceptors of Usher syndrome along with other mature retinal cell types, chronologically analogous to the development of the cell types in a mature human retina. This study also established a robust method for the in vitro derivation of RGCs and astrocytes from human iPS cells and provided novel methodologies and evidence to characterize these individual somatic cell types. Overall, this study provides a unique insight into the application of human pluripotent stem cell biology by establishing a novel platform for future studies of in vitro disease modeling of the retinal degenerative diseases: Usher syndrome and glaucoma. In downstream applications of this study, the disease relevant cell types derived from human iPS cells can be used as tools to further study disease progression, drug screening and cell replacement strategies.

Additionally, cyclosporin A, an inhibitor of the mitochondrial permeability transition (MPT) pore, failed to prevent hIAPP-induced DeltaPsim collapse, cytochrome c and AIF release and caspase-3 activation, indicating that the MPT pore was not involved in hIAPP-induced apoptosis. On the other hand, potential crosstalk between the extrinsic and intrinsic apoptotic pathways was demonstrated by cleavage of Bid by caspase-8 in the apoptotic process triggered by hIAPP.
It is widely accepted that human islet amyloid polypeptide (hIAPP) aggregation plays an important role in the loss of insulin-producing pancreatic beta cells. Insulin secretion impairment and cell apoptosis can be due to mitochondrial dysfunction in pancreatic beta cells. hIAPP-induced cytotoxicity is mediated by the generation of reactive oxygen species (ROS). Phycocyanin (PC) is a natural compound from blue-green algae that is widely used as food supplement. Currently, little information is available about the effect of hIAPP on mitochondrial function of beta cells and protection of PC against hIAPP-induced cytotoxicity. In this thesis, I hypothesize that hIAPP may impair beta cell function with the involvement of mitochrondrial dysfunction, and this effects could be attenuated by PC. Therefore, the aim of this study was to investigate the role of mitochondria in hIAPP-induced apoptosis, the in vitro protective effects of PC and explore the underlying mechanisms.
It was found that hIAPP induced apoptosis in INS-1E cells with the disruption of mitochondrial function, as evidenced by ATP depletion, mitochondrial mass reduction, mitochondrial fragmentation and loss of mitochondrial membrane potential (DeltaPsim). Further molecular analysis showed that hIAPP induced changes in the expression of Bcl-2 family members, release of cytochrome c and apoptosis-inducing factor (AIF) from mitochondria into cytosol, activation of caspases and cleavage of poly (ADP-ribose) polymerase. Interestingly, the hIAPP-induced mitochondrial dysfunction in INS1-E cells was effectively restored by co-treatment with PC.
Our results showed that hIAPP inhibited the INS-1E cell growth in a dose-dependent manner. However, cytotoxicity of hIAPP was significantly attenuated by co-incubation of the cells with PC. hIAPP induced DNA fragmentation and chromatin condensation, which were key characteristics of cell apoptosis. These changes were inhibited by PC as examined by TUNEL assay and DAPI staining. Moreover, PC significantly prevented the hIAPP-induced overproduction of intracellular ROS and malonaldehyde (MDA), as well as changes of activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) enzymes. Furthermore, hIAPP triggered the activation of mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinase (JNK) and p38 kinase, and these effects were effectively suppressed by PC.
Taken together, I have demonstrated for the first time the involvement of mitochondrial dysfunction in hIAPP-induced INS-1E cell apoptosis, which was attenuated by PC through attenuating oxidative stress, modulating JNK and p38 pathways and reducing mitochondrial dysfunction.
Li, Xiaoling.
Adviser: Juliana Chung Ngor Chan.
Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 150-159).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.