Dissertations / Theses on the topic 'Buxus'

Les travaux de recherche concernent la découverte d'inhibiteurs puissants et sélectifs de l'acétylcholinestérase (AChE) pouvant prévenir ou traiter les troubles de la mémoire associés au vieillissement ou à la maladie d'Alzheimer. Ces composés sont préparés de façon originale à partir d'un alcaloïde stéroïdique, isolé du buis des Baléares. Au cours d'un ciblage systématique des molécules de la chimiothéque de l'Institut de Chimie des Substances Naturelles nous avons identifié un puissant inhibiteur de l'acétylcholinestérase. Cette oxazine est issue de la transformation chimique d'un alcaloïde, la N-3-isobutyrylcycloxobuxidine-F isolée du buis des baléares (Buxus balearica Willd) selon un nouveau protocole. Les alcaloïdes bruts ont été fractionnés par extraction à différents pH en milieu tamponné. Ainsi, l'hémisynthèse de la tête de série a été réalisée. De nombreux analogues de l'oxazine ont été préparés de façon originale à partir du précurseur naturel pour établir des relations structure-activité. Ces synthèses ont mis enjeu de nombreuses réactions : réduction, ouverture contrôlée du cyclopropane, réaction de Polonovski " non classique " et amination réductrice. Dans le cas de la synthèse des tétrahydro-pyrimidines, les étapes de synthèse ont nécessité de nombreuses adaptations dues à la présence de l'atome d'azote à la place de l'oxygène oxazinique. Les relations structure-activité ont révélé l'importance de la présence du cycle oxazine ou pyrimidine dans la structure des inhibiteurs, ainsi que la présence du cyclopropane. Des différences d'activité des composés ont été observées en fonction de la source de l'AChE
These investigations aim at discovering powerful and selective inhibitors of acetylcholinesterase able to prevent or cure memory problems linked to aging or Alzheimer disease. During a systematic screening of the chemical library of the "Institut de Chimie des Substances Naturelles ", we identified a new hit as a potent inhibitor of acetylcholinesterase. This oxazine was obtained from the chemical transformation of an alkaloid, N-3-isobutyrylcycloxobuxidine-F extracted from the " Baléares Boxwood " (Buxus balearica Willd. ) using a new process. All the constituent alkaloids were fractionated by solvent and pH adjustments. Thus, hemisynthesis of the hit was achieved. Many novel synthetic analogues of the hit were then prepared from the natural precursor in order to establish structure-activity relationships. These original syntheses are based on several reactions : reduction, selective ring opening of a cyclopropane, " non-classical" Polonovski reaction and reductive animation. Concerning the tetrahydro-pyrimidine syntheses, the various steps required many adaptations due to the presence of the nitrogen atom instead of the oxazinic oxygen atom. The structure-activity relationships revealed the importance of the presence of the oxazin or pyrimidine rings in the inhibitor structures as well as the presence of the cyclopropane ring. Some differences in activity of the compounds were observed depending on the natural source of AchE

Ce travail concerne l'analyse de la preparation vegetale de buxus sempervirens administree aux patients dans le cadre d'un essai pilote phase i/ii, mis en place par l'equipe du professeur dellamonica en juin 1992 a l'hopital de nice. Nous avons mis au point une premiere methodologie de fractionnement impliquant un procede d'extraction suivi d'un traitement par chromatographie liquide qui nous a permis d'obtenir une fraction presentant in vitro a la fois une activite antivirale anti-vih et une activite inhibitrice de la production d'il2 interessante. Cependant, nos essais de reproductibilite n'ont pas abouti. Des essais de traitement, effectues en collaboration avec les laboratoires arkopharma indiquent une augmentation de l'activite biologique correlee avec le pourcentage d'alcaloides dans les extraits. Aussi, nous avons prepare un extrait d'alcaloides totaux que nous avons fractionne suivant une methodologie basee sur la difference de basicite des alcaloides en realisant des extractions a differents ph (8-7-6-5-4-3-2). Nous avons ainsi obtenu des fractions constantes dans leurs activites biologiques : faible activite anti-vih, activite inhibitrice notable de la production d'il2. La chromatographie liquide sur gel de silice ou d'alumine de ces fractions nous a permis d'isoler trois alcaloides deja decrits (la (-) 16--hydroxybuxaminone, la buxamine e et le buxaminol e) et quatre alcaloides originaux dont nous avons elucide la structure. Ces composes presentent un squelette 9,10-seco-buxa-9(11), 10(19)-diene, aussi nous les avons nommes : le n 2 0-formylbuxaminol e, le o 1 6-syringoylbuxaminol e, la n 2 0-acetylbuxamine e et la n 2 0-acetylbuxamine g. Dans le but de definir la composition chimique des extraits d'alcaloides totaux de buxus sempervirens, nous avons mis au point une methode d'analyse par couplage clhp-ms qui nous a conduit a l'identification de quinze constituants d'extraits d'alcaloides totaux, a la detection de tous les composants de ces extraits, a l'obtention de spectres uv et a la determination de masse moleculaire de chacun des solutes elues en une seule analyse. Ces travaux constituent la premiere etude de la composition chimique des alcaloides totaux de buxus sempervirens par chromatographie et permettent a la fois d'etablir des profils chromatographiques et de determiner les constituants des extraits a analyser.

L’objectif des travaux présentés dans ce manuscript a été la recherche d’inhibiteurs puissants et sélectifs de l’acétylcholinestérase (AChE) pouvant prévenir ou traiter les troubles de la mémoire et ralentir le développement des plaques de protéine β-amyloïde associées à la maladie d’Alzheimer. Au cours du criblage des molécules de la chimiothèque de l’Institut des Substances Naturelles, un puissant inhibiteur de l’AChE a été identifié. De nombreux analogues ont été obtenus par hémi-synthèse à partir d’un alcaloïde stéroïdien, la N-isobutyrylcycloxobuxidine-F, extraite du Buxus balearica. Les recherches se sont tout d’abord orientées sur la synthèse de nouveaux pharmacophores, d’une part au niveau du cycle dihydro-4H-[1,3]oxazine, et d’autre part au niveau de l’amine tertiaire par déméthylation puis acylation ou alkylation. Les synthèses ont nécessité de nombreuses adaptations à cause de la réactivité particulière des alcaloïdes stéroïdiens et surtout celle de l’alcool en position néopentylique. Les activités anticholinestérasiques ont été évaluées sur les AChEs humaine et d’Electrophorus electricus ainsi que sur la butyrylcholinestérase humaine. Les inhibiteurs les plus puissants ont démontré leur capacité à diminuer l’agrégation de la protéine β-amyloïde induite par l’AChE. Les études de relations structure-activité ont révélé l’importance de la présence du cycle dihydro-4H-[1,3]thiazine, du groupement isopropyle, du cyclopropane ainsi que d’un groupement benzyle positionné sur l’atome d’azote Nb
These investigations aim at the discovering of powerful and selective acetylcholinesterase (AChE) inhibitors able to prevent or cure memory problems and aggregation of β-amyloid protein linked to aging or Alzheimer disease. During a systematic screening of the chemical library of the “Institut de Chimie des Substances Naturelles”, a potent inhibitor of AChE was discovered. Many analogues have been obtained by hemisynthesis from a steroidal alkaloid, N isobutyrylcycloxobuxidine-F, extracted from Buxus balearica. First, research have been focused on the synthesis of new pharmacophores, at the dihydro-4H-[1,3]oxazine cycle and at the tertiary amine by demethylation followed by acylation or alkylation. Synthesis have required many adaptations due to the particular reactivity of the steroidal alcaloids and of the neopentylic alcohol. Anticholinesterasic activities have been evaluated on human and Electrophorus electricus AChES and on the human butyrylcholinesterase. The most powerful inhibitors have demonstrated their ability to decrease the aggregation of β-amyloid protein induced by AChE. The structure-activity relationship studies have showed the importance of the presence of the dihydro-4H-[1,3]thiazine cycle, of the isopropyl group, of the cyclopropane and of a benzyl group positioned on the nitrogen atom NB

Depuis les années 1990, on observe une accélération de la vitesse de propagation après établissementd’une grande partie des insectes introduits accidentellement en Europe. L’invasion fulgurante de lapyrale du buis, Cydalima perspectalis, est un exemple de ce phénomène. Observée pour la premièrefois en 2007 en Allemagne, elle s’est ensuite propagée jusqu’au Caucase en une décennie. Il a étérapidement supposé que le commerce de buis d’ornement entre les pays européens et la Chine auraitpermis son introduction tandis que l‘intensification des échanges commerciaux entre pays européensaurait facilité sa propagation à travers le continent. Cependant, certains traits biologiques de l’insecteont également pu favoriser son invasion. Pour mieux comprendre les facteurs impliqués dans soninvasion fulgurante, une approche pluridisciplinaire a été utilisée au cours de cette thèse. Tout d’abord,les routes d’invasion ont été appréhendées avec deux types de marqueurs génétiques. Puis, deux traitsbiologiques ayant pu favoriser son établissement et son expansion ont été étudiés : ses capacités devol et la diapause. Les résultats génétiques ont permis de cibler la partie côtière de la Chine commezone source probable, et suggèrent au moins deux introductions indépendantes depuis la Chine suivipar un phénomène tête de pont probablement lié au commerce d’ornement. Les capacités de volestimées sont élevées et expliqueraient la dispersion de l’insecte à l’échelle locale. De plus, la sortie dediapause influencée par la température a pu permettre l’adaptation du nombre de générations enfonction du climat annuel. Ces deux traits biologiques ont par conséquent dû favoriser son invasion àl’échelle locale mais il apparaît que les activités humaines, en particulier le commerce des plantesornementales, ont joué un rôle majeur dans son invasion fulgurante à large échelle en Europe et auCaucase
Since 1990, recently established non-native insect species appear to be spreading across Europesignificantly faster than before. The box tree moth, Cydalima perspectalis, is a representative of such arapid spread. First observed in 2007, it spread across Europe to Asia Minor in a decade, damaging onBuxus plants in urban areas as well as in forests. The trade of ornamental box plants between Europeand China has been hypothesized as the invasion pathway of the insect while the trade betweenEuropean countries could explain its fast expansion. The biological traits of the moth may also havefavored its spread. To understand the factors involved in both its invasion and fast expansion, we useda multidisciplinary approach. First, the invasion routes were tentatively assessed using two geneticmarkers. Then, two biological traits, flight capacities and diapause, were studied because they may havepromote its spread. The genetic analyses clearly suggest an origin from Eastern China origin but alsocomplex invasion pathways. Two independent introduction events probably happened directly fromChina followed by bridgehead effects due to ornamental plant trade. The moth flight capacities areimportant but can only explain dispersal at local scale. Moreover, diapause termination was influencedby temperature, which may lead to an adapted number of generation depending on local climate. Thesebiological traits may have favored local invasions. However, our study suggests that human-mediateddispersal played a key role in the fast expansion of C. perspectalis across Europe and Asia Minor

Pour lutter contre l'invasion des pelouses calcicoles par des especes ligneuses, des mesures agri-environnementales preconisent l'utilisation du paturage. Leur efficacite a ete evaluee en etudiant la regeneration de buxus sempervirens et de quercus humilis. Parmi les mecanismes qui affectent ce stade de vie, l'accent est mis sur les interactions positives entre plantes dont le caractere obligatoire dans les successions vegetales est rarement mis en evidence. Les individus de buxus, non consommes par les ovins au stade adulte, sont broutes, en proportion importante, au stade semis. Mais la presence de plantes peu palatables, autour de certains semis, facilite indirectement leur installation en les protegeant du paturage. Le transport des graines de buxus par les fourmis, dans ce type de voisinage, peut favoriser indirectement la survie des semis mais aussi, augmenter de trois fois la distance de dissemination maximum apres projection balistique. Une fois adultes, les individus de buxus conferent une protection contre le paturage aux semis de quercus installes sous leur canopee. Ils ameliorent egalement les conditions de germination et reduisent la mortalite estivale. Mais, ils ont aussi des effets negatifs en reduisant la ressource en lumiere et en augmentant indirectement la predation par les rongeurs. Un modele des effets du voisinage sur la distribution des semis des especes ligneuses, en fonction de l'intensite de paturage, est propose. Les intensites de paturage moyennes favorisent une structuration spatiale des ligneux. Meme a des intensites elevees, la presence de plantes non consommees par les ovins permet indirectement la fermeture des paysages en facilitant l'installation des ligneux. Les interactions positives entre plantes peuvent jouer un role considerable dans la coexistence des especes et dans la dynamique de la vegetation.

Scholars have faced many challenges in the classification of the literary genre of the hagiographical texts. In addition to their various styles and structures, hagiographical texts tend to move beyond the classical rhetorical approach. So, it is preferable to regard hagiography as a discourse which was mainly written for the purpose of the production of new heroes through the imitation of Christ and His holy men/women. The hagiographical discourse continued in Early Medieval Arabic Christianity, yet its purpose has expanded to address both Christians and Muslims. Through the examination of the Arabic hagiographical texts of two neo-martyrs, Anthony al-Qurashi and Būlus ibn Raja, it has been revealed that Christians pursued the figure of speech of parrhesia to address the mixed audience. On the one hand, to urge Christians to behold to their faith and, on the other hand, to encourage Muslims to convert.

Danggui Buxue Tang (DBT), a herbal decoction containing Astragali Radix (AR) and Angelicae Sinensis Radix (ASR), has been used in treating menopausal irregularity in women for more than 800 years in China. Pharmacological results showed that DBT exhibited significant estrogenic properties in vitro, which therefore suggested that DBT could activate the nuclear estrogen receptors. Here, we assessed the estrogenic properties of DBT in an ovariectomized in vivo rat model: DBT was applied to the ovariectomized rats for 3 days. The application of DBT did not alter the weight of uterus and liver, as well as the transcript expression of the proliferation markers including the estrogen receptors α and β. However, DBT stimulated the transcript expression of the estrogen responsive genes. In addition, the inductive role of DBT on the expression of members of the aryl hydrocarbon receptor family in uterus and liver of ovariectomized rats was confirmed. These responses of DBT however were clearly distinct from the response pattern detectable here for 17β-estradiol. Therefore, DBT exhibited weak, but significant, estrogenic properties in vivo; however, some of its activities were independent of the estrogen receptor. Thus, DBT could be an exciting Chinese herbal decoction for an alternative treatment of hormone replacement therapy for women in menopause without subsequent estrogenic side effects.

Danggui Buxue Tang (DBT), a herbal decoction containing Astragali Radix (AR) and Angelicae Sinensis Radix (ASR), has been used in treating menopausal irregularity in women for more than 800 years in China. Pharmacological results showed that DBT exhibited significant estrogenic properties in vitro, which therefore suggested that DBT could activate the nuclear estrogen receptors. Here, we assessed the estrogenic properties of DBT in an ovariectomized in vivo rat model: DBT was applied to the ovariectomized rats for 3 days. The application of DBT did not alter the weight of uterus and liver, as well as the transcript expression of the proliferation markers including the estrogen receptors α and β. However, DBT stimulated the transcript expression of the estrogen responsive genes. In addition, the inductive role of DBT on the expression of members of the aryl hydrocarbon receptor family in uterus and liver of ovariectomized rats was confirmed. These responses of DBT however were clearly distinct from the response pattern detectable here for 17β-estradiol. Therefore, DBT exhibited weak, but significant, estrogenic properties in vivo; however, some of its activities were independent of the estrogen receptor. Thus, DBT could be an exciting Chinese herbal decoction for an alternative treatment of hormone replacement therapy for women in menopause without subsequent estrogenic side effects.

Dissertação de Mestrado em Engenharia do Ambiente
Buxus é um género exótico introduzido na Península Ibérica (PI) pelas dinâmicas glaciares. Caracterizado por arbustos de folha persistente, o género apresenta aspetos morfológicos peculiares e uma vasta gama ecológica, o que o torna economicamente importante. Existem apenas duas espécies de Buxus presentes na Europa, Buxus sempervirens e Buxus balearica. A primeira encontra-se distribuída pelo centro e sul da Europa, e em Portugal apresenta maior concentração em Trás-os-Montes, nos vales do Rio Sabor e Tua. Por sua vez, B. balearica mostra uma distribuição limitada às Ilhas Baleares e ao sul de Espanha. No presente estudo pretendeu-se caracterizar e estabelecer aspetos que contribuam para a distinção das duas espécies, e determinar variações dentro da mesma espécie em resultado de adaptações às condições edafo-climáticas das regiões. Foi realizado um estudo morfo-anatómico em amostras foliares de exemplares respetivos a 18 populações de Buxus, oriundos da Península Ibérica: 6 populações de B. balearica, 6 de B. sempervirens do Sistema Cantábrico (SC) e 6 de B. sempervirens do Sistema Ibérico (Ib). Este estudo incluiu a análise da anatomia foliar e de aspetos morfo-anatómicos relacionados com a economia da água nas folhas. Verificou-se que as espécies apresentam diferenças morfo-anatómicas. A espécie B. balearica apresenta área foliar e massa por unidade de área da superfície foliar (LMA) superiores, ao passo que B. sempervirens possui menores folhas com menor espessura foliar e menor LMA. Ambas as espécies possuem folhas dorsiventrais e hipoestomáticas, embora B. balearica apresente maior densidade de estomas. Apresentam também características xeromórficas, indicando estar adaptadas à secura. As populações de B. sempervirens do Sistema Ibérico parecem adotar mecanismos de resistência ao clima Mediterrânico, tais como maior espessura da folha, do parênquima clorofilino em paliçada e da cutícula, bem como LMA e densidade estomática superiores. Pela análise canónica discriminante, a espessura da cutícula superior foi a variável principal na discriminação de todas as populações de Buxus. As variações morfo-anatómicas entre as várias populações estudadas podem ser o resultado das variações regionais no ambiente, tais como a fertilidade do solo, sazonalidade térmica e/ou diferenças filogenéticas.
Buxus is an exotic genus introduced in the Iberian Peninsula by glacial dynamics. This genus is characterized by evergreen shrubs having some distinctive features and a wide range of ecological characteristics, which makes it economically important. There are only two species of Buxus in Europe, Buxus sempervirens and Buxus balearica. The first one is extensively distributed along Central Europe, and in Portugal it is found in Trás-os-Montes in the valley of Sabor River and Tua. In turn, B. balearica has a distribution limited to the Balearean Island and south of Spain. The objective of the present study was to characterize and establish relevant aspects that contribute to the distinction of the two species, and to determine the variations within the same species related to the adaptation to the environmental conditions of the regions. A morpho-anatomical study was carried out using samples of leaves from 18 populations of Buxus, gathered in the Iberian Peninsula: 6 populations of B. balearica, 6 of B. sempervirens from the Cantabrian System and 6 of B. sempervirens from the Iberian System. This study included the leaf anatomical analysis and morpho-anatomical aspects related to the way the leaves avoid dehydration. The obtained results indicate morpho-anatomical differences between the species. B. balearica has bigger foliar area and higher leaf mass per unit area (LMA) than B. sempervirens that has smaller and thinner leaves, with lower LMA. Both species contain dorsiventral and hypostomatic leaves, although B. balearica presents higher stomatal density. Moreover, these two species present xeromorphic features that indicate good adaptation to drought. The population of the Iberian System of B. sempervirens revealed mechanisms of resistance to the Mediterranean climate such as thicker leaves, in consequence of thicker palisade parenchyma and cuticle, as well as higher LMA and higher stomatal density. The discriminant canonical analysis shows that the upper cuticle thickness was the principal morpho-anatomical variable for the discrimination of all the populations of Buxus. The morpho-anatomical variations from all the studied populations can be the result of the regional variations of the environment, such as the soil fertility, thermal seasonality and/or phylogenetic differences.

This thesis describes phytochemical studies on Buxus macowanii, a medicinal plant collected from South Africa and the fungal transformation of glycyrrhetinic acid (71), a triterpenoid found in Glycyrrhiza plants. The crude methanolic extract of B. macowanii was active in the acetylcholinesterase inhibition assay (IC50 = 30 μg/mL). Chemical investigation of its alkaline dichloromethane extract afforded five novel triterpenoidal alkaloids, 31-hydroxybuxatrienone (56), macowanioxazine (57), 16α-hydroxyma- cowanitriene (58), macowanitriene (59), and macowamine (60), along with two known Buxus bases, Nb-demethylpapillotrienine (61) and moenjodaramine (62). Compounds 56, 57, and 61 exhibited different level of AChE inhibitory activities with compound 56 being significantly active (IC50 = 8.0 μM). Glycyrrhetinic acid (71) was incubated with Curvularia lunata (ATCC 12017) for ten days to afford one metabolite which was characterized by NMR spectral data as known biotransformed product, 3-oxo-glycyrrhetinic acid (85). This metabolite was previously obtained through biotransformation of 71 by using other fungi.

This thesis describes phytochemical studies on two medically important plants, Buxus natalensis and Drypetes gossweileri. Chemical investigation on the acetylcholinesterase inhibiting chloroform extracts, obtained at pH 7.0 and 9.5 resulted in the isolation of seven natural products: O10-natafuranamine (123), cyclonataminol A (124), 31-demethylbuxaminol A (125), buxaminol A (126), buxaminol C (127), p coumaroylputrescine (128) and methyl syringate (129). Compound 123 is a member of a rarely occurring class of Buxus alkaloids containing a tetrahydrofuran ring incorporated in its structure. Compounds 123-129 were isolated for the first time from this plant. Structures of compounds 123-129 were elucidated with the aid of NMR and MS spectral data. All of these isolates exhibited different levels of AChE inhibitory activities with compound 123 being significantly active in this bioassay with an IC50 of 8.5 µM compared to the rest of the isolates. Compounds 123-129 were inactive in antimicrobial assays. Phytochemical studies on the crude extract of Drypetes gossweileri resulted in the isolation of a new N-linked aromatic glycoside, N-β-glucopyranosyl-p-hydroxy phenyl acetamide (151), along with two known compounds, p-hydroxy phenyl acetic acid (152) and p hydroxyphenyl acetonitrile (153). Compounds 151-153 exhibited moderate to weak AChE inhibitory activities.

This thesis describes the isolation and characterization of four steroidal alkaloids from Buxus macowanii. Phytochemical investigation of Buxus macowanii resulted in the isolation of two novel steroidal alkaloids, Nb-demethyl-6-deoxy-16-acetoxy O2-natafuranamin (112), and 6-deoxy-16-acetoxy O10-natafuranamin (113) alongside two known steroidal alkaloids, cycloprotobuxine-D (114), and cycloprotobuxine-F (115). Compounds 114 and 115 have been isolated for the first time from B. macowanii. Structure of compounds 112-115 was elucidated with aid of UV, IR, mass, and 1D and 2D NMR spectroscopy. These compounds showed different level of anti-AChE activity. Among all the isolates, compound 112 was found to be significantly active against AChE with an IC50 value of 4.7 µM. The bioactivity of this new compound nearly comparable to those of huperzine (IC50 = 1.7 µM) and O2-natafuranamine (IC50 = 3.0 µM).

碩士
國立臺灣大學
生態學與演化生物學研究所
94
Floral organ determination is best explained by the ABCDE model postulated by genetic studies of Arabidopsis thaliana. Sepals are determined by A and E class genes; petals are determined by A, B, and E; stamens by B, C, and E; and carpels by C and E class genes. A, B, C, and E class gene lineages are known having duplicated several times during the evolution of angiosperms. One of the noted major duplication events occurred in the origin of the early angiosperms, leading to the formation of subgroups of B/C/D/E class. Another one occurred near the basal eudicots and gave rise to further subgroups in A/B/C/D/E class genes among core eudicots. The phylogenetic position of the family Buxaceae is located right where the second major duplication of ABCDE genes might have occurred, which is supported by multiple gene (nuclear 18S rDNA, chloroplast rbcL and atpB) phylogenetic analyses. Therefore, the identification and characterization of floral organ identity genes in Buxaceae is critical in elucidating the gene evolution related to floral organ formation. Floral organ identity gene homologues of Buxus microphylla ssp. sinica were screened, and nine gene homologues of B. microphylla ssp. sinica were successfully identified in this study. Two of them are A class gene homologues, four B class, two C class, and one E class. Expression patterns based on RT-PCR of floral organ identity genes of Buxus do not match with current ABCDE model. There is no expression of all identified genes in leaves, and all genes are expressed in all floral parts except BumPI2. BumPI2 does not express in pistillodes, and only weakly expresses in stamens. Whether or not the discrepancy is due to a deviation from the standard floral model awaits further examinations. According to the phylogenetic analyses, the homologous genes of Buxus belong to their expected clades, showing a position near the base of core eudicots. Genes of Buxus are always sister to genes of Pachysandra or Trochodendron,and this clade is close to core eudicots in all analysis. BumAP3 genes are the only exception that sometimes grouped with Pachysandra genes as sister group to euAP3 clade. Moreover, frameshift modulation of BumAP3-1/2/3 genes indicates their high frameshift potential and frameshift mutation might be the key mechanism occurred after duplication event and led to diversification of core eudicots.

碩士
國立臺灣大學
森林學研究所
88
Abstract Buxus liukiuensis ,a rare plant of Taiwan and Lyukyus ,usually occurs at eastern and southern parts of Taiwan ,and its elevations ranges form 100 to 1,500 meters above sea level, at limestone sites:they occupy a small isolated patches mixed with other trees in forests. This aim of this study is to research the genetic variation of Buxus liukiuensis Makino. by RAPDs analysis : a total of 150 samples of Buxus liukiuensis which are representing the populations of eight area were collected.A test of random amplified polymorphic DNA to identify its genetic variation .and Amplification with 9 primers to yield a total of 97 polymorphic RAPDs bands were recorded.Among them ,92 bands are polymorphism(94.85﹪),and 8 DNA bands are monomophism(8.25﹪). A UPGMA cluster analysis was performed on the Dice''s similarity matrix which derives form eight populations of plants by Ntsys. Partition the genetic variation into within and between populations of Buxus liukiuensis through POPGENE analysis indicated 36.54﹪of variation is between populations while63.46﹪of variation within population.It is suggested,based on the studies,there is not or lower gene interflow existed among populations,indicating great genetic differentiation which was mostly caused by geographical isolation occurred.

碩士
臺北醫學大學
生藥學研究所
98
Iron deficiency anemia, the most common type of anemia, is caused by reasons such as iron malabsoption, iron-deficient diet, and blood loss. The traditional Chinese medicine, Danggui Buxue Tang (DBT), containing Radix angelicae sinensis (Danggui) and Radix astragali (Huangqi) has been used to invigorate “Qi” and nourish “Blood”. In this study, DBT was combined with iron to treat iron deficiency anemia. Another objective was to delineate whether the “nourish the blood”, as described in the ancient medicine books, were related to the iron deficient anemia treatment or not. In the first place, Caco-2 cell iron uptake model was used to detect the effect of ferritin biosynthesis. The results showed that Danggui and Huangqi could not enhance ferritin biosynthesis, but DBT could significantly promote ferritin biosynthesis. The principle constituent of DBT, ferulic acid, also could enhance iron absorption. The treatment program of DBT and iron in Caco-2 cells were exchanged to know the interaction of DBT and iron. The results revealed while Caco-2 cells were co-treated with DBT and iron, DBT could inhibit the absorption of iron. When DBT or iron was pre-treated, DBT could significantly enhance the absorption of iron. Therefore, the rat hemoglobin repletion assay was used to explore the above therapeutic activity in vivo. The result showed that the absorption of iron was reduced after co-treatment with DBT and iron in Wistar rats. However, the hemoglobin level was increased in the DBT group. The bioavailability of the iron was increased up to 1947% in DBT group in the hemoglobin repletion efficiency (HRE) test. The results of phytochemical analysis revealed that there are phenol compounds (6.51mg/g), condensed tannins (2.31mg/g), and saponins compounds (34.48mg/g) in DBT. In addition, the total ferrous content was decreased in the mixture of DBT and iron, indicating of that certain components of DBT like phenols, condensed tannins could chelate the iron. Besides, DBT could activate the DMT-1 expression in Caco-2 cells by Western blotting assay. Therefore, we assumed the iron promoting absorption of DBT was mediated through the activation the DMT-1 in Caco-2 cells. In conclusion, the DBT enhanced the bioavailability of the iron both in vitro and in vivo. The absorption of iron was inhibited by DBT and iron co-treatment, but enhanced by DBT and iron pre-treatment. Based on the above results, we suggested DBT and iron should be administrated separately for iron deficient anemia patients.

碩士
國立中興大學
生命科學系
93
Colorectal cancer is the third most common cancer and the fourth most frequent cause of cancer deaths worldwide. The primary treatment of colon cancer is the surgical resection, and chemotherapy treat late stage colorectal cancer. The cancer patients may suffer from decrease of immunity after chemotherapy. A lot of cancer patients will take Chinese herb during or after chemotherapy. In recent years, Danggui Buxue Tang (DGBXT) has been proven to promote the production of red blood cell, to enhance the immune function and to inhibit tumor growth. In this study, we try to investigate the anti-cancer activity of DGBXT on colon cancer cell line, CT-26 and HCT-8. We also try to investigate the influence of the decoction on colon cancer animal model. Our data show that dose and time dependent effects of DGBXT inhibited colon cancer cell line proliferation. We analyzed the cell population during cell cycle by flow cytometry, and demonstrated that DGBXT triggered the arrest of cell cycle in G0/G1 phase. Moreover DGBXT induced apoptosis in colorectal cell line CT-26 that measured by TUNEL staining and Annexin V-FITC/PI stain. Cell viability and cell population analysis performed on CT-26 cell line in comparison to three known compounds in DGBXT - ferulic acid, formononetin and Astragaloside IV. Cell viability showed the inhibition rates of each individual compound was within 15%-25% at the concentration of 10-4M. The inhibition rate of the complex contained these above compounds was 46%. Cell population analysis demonstrated that ferulic acid, formononetin and Astragaloside IV triggered the arrest of cell cycle in G0/G1 phase and increased percentage of sub-G1 phase dose and time dependent. The Balb/C mice bearing CT-26 cell line was used as animal model to study the effect of DGBXT on colon cancer. All mice divided into 4 groups as Group I: Feeding normal saline as control group; Group II: Feeding DGBXT; Group III: injecting 5-FU; Group IV: Feeding DGBXT and injecting 5-FU. Experiment results demonstrated that DGBXT significant inhibited tumor growth. Compared the 5-FU treatment with 5-FU combined DGBXT treatment, the toxicity and side effects such as leukopenia and weight loss which caused by 5-FU treatment were reduced by DGBXT. There were no significant changes in body weight, hepatic and renal functions in the group of mice fed DGBXT.

Exame público realizado em 22 de Julho de 2015.
Dissertação de mestrado realizada no âmbito do Mestrado em Arquitectura.
O tema abordado nesta dissertação é a Metamorfose da Forma como instrumento de estudo e leitura crítica do projeto de Arquitetura. Neste sentido, procura-se estudar através do caso de estudo do Solar da Quinta dos Buxos uma integração multidisciplinar de arquiteturas, da forma e sua metamorfose em ritmos como peça integrante de um todo formal de modo a dar uma resposta positiva e inclusiva num desenho arquitetural. Com o desenvolvimento deste trabalho, perspetiva-se um estudo mas, também, deixar em aberto um campo na área da investigação de estruturas de apoio contemporâneas a arquiteturas vernáculas independentemente do seu estado de conservação, do seu programa prévio em arquiteturas de apoio e seu âmbito multidisciplinar de modo que a forma seja uma metamorfose que gera um ritmo de constantes assemblagens entre si numa linguagem própria, criando um corpo arquitetural de leitura como um todo contínuo válido e coerente. Por último, no caso de estudo realizado, foi de importante todo o contexto de isolamento em que o mesmo se insere, para uma constante procura de diálogos e aprendizagens dos mesmos no ato de desenho e concepção, sendo estes os valores que norteiam o objectivo deste estudo, valores que revelam o conceito. Importa realçar que se procura tomar a recuperação do património numa vertente sustentável e dotar o mesmo de um aspeto formal, que o torne integrante numa época e funções contemporâneas, sem adulterar e/ou perder a sua génese. Abstract: The topic of this dissertation is the Metamorphosis of Form, as this study tool and critical reading of the architecture project. In this sense, looking to study through the case study of the solar Quinta dos Buxos a multidisciplinary integration architectures, form and its metamorphosis into rhythms as part of a formal integral whole in order to make a positive and inclusive response in architectural design. With the development of this work, we try to study and leave an open field in research of contemporary support structures the vernacular architectures regardless of their condition, its prior program in support of architectures and its multidisciplinary scope to the form is a metamorphosis that generates a constant rate of assemblies together in a language itself, generating a read Architectural body as a whole still valid and consistent. Finally, the case study and taken, was important all the isolation of the context in which it is situated, for a constant search for dialogue, and learning from them in the act drawing and design, which are the values that prompt the aim of this study these are the values that reveal the concept. It should be noted that seeks to take the recovery of assets in a sustainable present, and provide the same formal aspect that makes it integral at a time and contemporary features and without misrepresenting or lose its genesis.
Abstract: The topic of this dissertation is the Metamorphosis of Form, as this study tool and critical reading of the architecture project. In this sense, looking to study through the case study of the solar Quinta dos Buxos a multidisciplinary integration architectures, form and its metamorphosis into rhythms as part of a formal integral whole in order to make a positive and inclusive response in architectural design. With the development of this work, we try to study and leave an open field in research of contemporary support structures the vernacular architectures regardless of their condition, its prior program in support of architectures and its multidisciplinary scope to the form is a metamorphosis that generates a constant rate of assemblies together in a language itself, generating a read Architectural body as a whole still valid and consistent. Finally, the case study and taken, was important all the isolation of the context in which it is situated, for a constant search for dialogue, and learning from them in the act drawing and design, which are the values that prompt the aim of this study these are the values that reveal the concept. It should be noted that seeks to take the recovery of assets in a sustainable present, and provide the same formal aspect that makes it integral at a time and contemporary features and without misrepresenting or lose its genesis.

博士
長庚大學
臨床醫學研究所中醫組
105
Chemotherapy is an important treatment modality for colon cancer, and concurrent chemoradiation therapy (CCRT) is the preferred treatment route for patients with stage II and III rectal cancer. The induction of autophagic cell death is an important process in the development of anticancer therapeutics. We examined how could DangGui BuXue Tang (DBT), a traditional Chinese herbal extract, against colorectal cancer (CRC) and its autophagy-related mechanism in sensitizing colorectal cancer cells for anticancer treatments. The polysaccharide-depleted fraction of DBT (DBT-PD) contains greater amounts of astragaloside IV and ferulic acid than the original formula does. Treatment of the murine colon carcinoma cell line (CT26) with DBT-PD inhibited cell growth, whereas treatment with comparable amounts of purified astragaloside IV and ferulic acid showed no significant effect. Concurrent treatment with DBT-PD increases the growth inhibitory effect of 5-fluorouracil up to 4.39-fold. DBT-PD enhances the effect of radiation therapy (RT) with a sensitizer enhancement ratio (SER) of up to 1.3 fold. It also increases the therapeutic effect of CCRT on CT26 cells. Cells treated with DBP-PD showed ultrastructural changes characteristic of autophagy, including multiple cytoplasmic vacuoles with double-layered membranes, vacuoles containing remnants of degraded organelles, marked swelling and vacuolization of mitochondria, and autolysosome-like vacuoles. In the tumor specimen, the expression of microtubule-associated proteins 1A/1B light chain 3B (LC3B) was upregulated by DBT-PD and DBT. In vitro experiments for mechanism clarification demonstrated that DBT-PD could induce autophagic death in CT26 cells accompanied by LC3B lipidation, downregulation of phospho-p70s6k, and upregulation of Atg7. RNA interference of Atg7, but not Atg5, partially reversed the effect of DBT-PD on LC3B lipidation and expression of phospho-p70s6k and Atg7. The changes in ultrastructural morphology and LC3B expression induced by DBT-PD were also partially blocked by the knockdown of Atg7 mRNA. We conclude that DBT-PD induces autophagy-associated cell death in CT26 cells through the upregulation of Atg7 and modulation of the mTOR/p70s6k signaling pathway, and may have the potential as the chemotherapy or radiotherapy sensitizer in colorectal cancer treatment.

Li Tin Wai Olive.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 191-217).
Abstracts in English and Chinese.
ACKNOWLEDGEMENTS --- p.II
ABSTRACT --- p.III
中文摘要 --- p.IX
PUBLICATIONS --- p.XIV
TABLE OF CONTENTS --- p.XV
LIST OF ABBREVIATIONS --- p.XXI
LIST OF FIGURES AND TABLES --- p.XXIII
Chapter CHAPTER 1. --- INTRODUCTION --- p.1
Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.7
Chapter 2.1. --- Cardiovascular disease --- p.7
Chapter 2.1.1. --- Introduction --- p.7
Chapter 2.1.2. --- Atherosclerosis --- p.7
Chapter 2.1.3. --- Cellular and molecular deregulation in early atherosclerosis --- p.10
Chapter 2.1.3.1. --- Introduction --- p.10
Chapter 2.1.3.2. --- Endothelial dysfunction --- p.11
Chapter 2.1.3.3. --- Nitric oxide --- p.12
Chapter 2.1.3.4. --- Adhesion molecules and the early events of atherogenesis --- p.13
Chapter 2.1.3.4.1. --- Introduction --- p.13
Chapter 2.1.3.4.2. --- Intracellular adhesion molecule-1 --- p.15
Chapter 2.1.3.4.3. --- Nuclear factor kappa B --- p.18
Chapter 2.1.3.5. --- Summary --- p.19
Chapter 2.2. --- Nitric oxide in molecular vascular biology --- p.20
Chapter 2.2.1. --- Introduction --- p.20
Chapter 2.2.2. --- Nitric oxide synthase --- p.21
Chapter 2.2.2.1. --- Introduction --- p.21
Chapter 2.2.2.2. --- Endothelial nitric oxide synthase --- p.24
Chapter 2.2.2.3. --- Inducible nitric oxide synthase --- p.25
Chapter 2.2.2.4. --- Nitric oxide concentration dependent effector pathways --- p.26
Chapter 2.2.3. --- Nitric oxide and its regulation in vascular events --- p.28
Chapter 2.2.3.1. --- Introduction --- p.28
Chapter 2.2.3.2. --- Regulation of vascular tone --- p.30
Chapter 2.2.3.3. --- "Regulation of platelet adhesion, activation and aggregation" --- p.32
Chapter 2.2.3.4. --- Regulation of endothelial adhesiveness and leukocyte adhesion - Anti-adhesive effect of nitric oxide --- p.32
Chapter 2.2.3.5. --- "Regulation of vascular smooth muscle growth, migration and proliferation" --- p.33
Chapter 2.2.3.6. --- Antioxidative effect of nitric oxide --- p.34
Chapter 2.2.3.7. --- Regulation of endothelial apoptosis --- p.35
Chapter 2.2.3.8. --- Nitric oxide and its relationship with other risk factors --- p.36
Chapter 2.3. --- "Menopause, cardiovascular diseases and Traditional Chinese Medicine" --- p.37
Chapter 2.3.1. --- Traditional Chinese Medicine and menopause --- p.37
Chapter 2.3.2. --- Dang Gui Buxue Tang --- p.38
Chapter 2.3.3. --- Danggui --- p.39
Chapter 2.3.3.1. --- Botanic origins --- p.39
Chapter 2.3.3.2. --- Usage --- p.39
Chapter 2.3.4. --- Huangqi --- p.40
Chapter 2.3.4.1. --- Botanic origins --- p.40
Chapter 2.3.4.2. --- Usage --- p.40
Chapter 2.3.5. --- Modern scientific research --- p.41
Chapter 2.3.5.1. --- General cardioprotective role --- p.41
Chapter 2.3.5.2. --- Vascular tone modulation --- p.42
Chapter 2.3.5.3. --- Haemostasis --- p.42
Chapter 2.3.5.4. --- Endothelial cell --- p.43
Chapter 2.3.5.4.1. --- Nitric oxide pathway --- p.43
Chapter 2.3.5.4.1.1. --- Direct alteration of nitric oxide secretion --- p.43
Chapter 2.3.5.4.1.2. --- Alteration of Nitric oxide synthase expression or activity --- p.43
Chapter 2.3.5.4.2. --- Alteration of adhesion molecule expression --- p.44
Chapter 2.3.5.4.3. --- Alteration of adhesion molecule expression as an effect of nitric oxide secretion --- p.45
Chapter 2.3.5.5. --- Antioxidant effect --- p.45
Chapter 2.3.5.6. --- Estrogenicity of DBT --- p.46
Chapter 2.4. --- Research plan --- p.47
Chapter 2.4.1. --- Formulation of research hypotheses --- p.47
Chapter 2.4.1.1. --- Hypotheses --- p.50
Chapter 2.4.2. --- Plan of study --- p.50
Chapter 2.4.2.1. --- Dang Gui Buxue Tang extraction and standardization of content --- p.50
Chapter 2.4.2.2. --- Cell model development --- p.52
Chapter 2.4.2.3. --- Experimental studies --- p.54
Chapter 2.4.2.4. --- eNOS activity determination - the nitric oxide metabolite assay --- p.56
Chapter 2.4.2.5. --- Endotoxin contamination in DBT --- p.57
Chapter 2.4.3. --- Sample size and statistical analysis --- p.59
Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.62
Chapter 3.1. --- Dang Gui Buxue Tang extraction and content standardization --- p.62
Chapter 3.1.1. --- Plant materials --- p.62
Chapter 3.1.2. --- DBT authentication --- p.62
Chapter 3.1.3. --- DBT processing prior to extraction --- p.63
Chapter 3.1.4. --- DBT extraction --- p.63
Chapter 3.1.5. --- Quantitative standardization of DBT markers by High Pressure Liquid Chromatography --- p.66
Chapter 3.1.5.1. --- DBT markers: standard preparation --- p.66
Chapter 3.1.5.2. --- Sample preparation --- p.67
Chapter 3.1.5.3. --- Quantitative analysis of DBT constituents by HPLC --- p.67
Chapter 3.1.6. --- DBT polysaccharide standardization --- p.68
Chapter 3.1.6.1. --- Glucose standard preparation --- p.68
Chapter 3.1.6.2. --- Sample preparation --- p.68
Chapter 3.1.6.3. --- Quantitative determination of polysaccharide by Phenol-Sulfuric acid colorimetric assay --- p.68
Chapter 3.1.7. --- DBT endotoxin contamination determination --- p.69
Chapter 3.1.7.1. --- "Positive, negative and inhibition controls" --- p.69
Chapter 3.1.7.2. --- Qualitative determination of sample endotoxin --- p.70
Chapter 3.2. --- Cell culture --- p.70
Chapter 3.2.1. --- Characterization of cultured cells --- p.72
Chapter 3.2.2. --- Passage --- p.73
Chapter 3.3. --- DBT treatment --- p.73
Chapter 3.3.1. --- Solvent system of DBT treatment --- p.73
Chapter 3.3.2. --- Dosage and duration of DBT treatment --- p.74
Chapter 3.3.3. --- Positive and negative controls --- p.74
Chapter 3.4. --- MTT-based cytotoxicity assay --- p.75
Chapter 3.5. --- Reverse transcriptase- polymerase chain reaction --- p.76
Chapter 3.5.1. --- Sample preparation --- p.76
Chapter 3.5.1.1. --- Total RNA isolation --- p.76
Chapter 3.5.1.2. --- DNase treatment --- p.77
Chapter 3.5.1.3. --- RNAethanol precipitation --- p.78
Chapter 3.5.1.4. --- Complementary DNA synthesis --- p.78
Chapter 3.5.2. --- Polymerase chain reaction --- p.79
Chapter 3.5.2.1. --- Polymerase chain reaction conditions --- p.79
Chapter 3.5.2.2. --- Primers --- p.79
Chapter 3.5.3. --- Visualization of the PCR products --- p.81
Chapter 3.5.3.1. --- Gel electrophoresis --- p.81
Chapter 3.5.3.2. --- Gel Doc software --- p.82
Chapter 3.5.3.3. --- Densitometry --- p.82
Chapter 3.5.4. --- Real time RT-PCR --- p.82
Chapter 3.6. --- Quantitative Immunocytochemical studies --- p.84
Chapter 3.6.1. --- Coverslip preparation --- p.84
Chapter 3.6.2. --- Sample preparation --- p.84
Chapter 3.6.3. --- Immunocytochemical staining preparation --- p.85
Chapter 3.6.3.1. --- "Immunocytochemical staining for vWF, α-actin, iNOS, ICAM-1, NF-kB using DAKO catalyzed signal amplification (CSA) system (anti-mouse)" --- p.86
Chapter 3.6.3.2. --- Immunocytochemical staining for eNOS using Santa Cruz immunoCruz staining system (anti-goat) --- p.87
Chapter 3.6.4. --- Counterstaining and mounting --- p.88
Chapter 3.6.5. --- Result interpretation --- p.89
Chapter 3.6.5.1. --- Microscopy and digital image capture --- p.89
Chapter 3.6.5.2. --- Determination of Image (file) Energy --- p.89
Chapter 3.7. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.90
Chapter 3.7.1. --- Sample preparation --- p.90
Chapter 3.7.2. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.91
Chapter CHAPTER 4. --- RESULTS --- p.93
Chapter 4.1. --- Dang Gui Buxue Tang extraction and standardization of content --- p.93
Chapter 4.1.1. --- DBT extraction - general data --- p.93
Chapter 4.1.2. --- DBT polysaccharide standardization --- p.97
Chapter 4.1.3. --- DBT marker standardization --- p.101
Chapter 4.1.4. --- DBT endotoxin contamination determination --- p.104
Chapter 4.2. --- Cell model development --- p.108
Chapter 4.2.1. --- Endothelial morphology --- p.108
Chapter 4.2.2. --- Immunocytochemistiy --- p.108
Chapter 4.2.3. --- MTT cytotoxicity assay --- p.110
Chapter 4.3. --- Study 1 --- p.112
Chapter 4.3.1. --- Immunocytochemistry (Hypothesis 1) --- p.112
Chapter 4.3.2. --- RT-PCR (Hypothesis 1) --- p.117
Chapter 4.3.3. --- Real time RT-PCR (Hypothesis 1) --- p.121
Chapter 4.3.4. --- Immunocytochemistry (Hypothesis 2) --- p.125
Chapter 4.3.5. --- RT-PCR (Hypothesis 2) --- p.128
Chapter 4.3.6. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.131
Chapter 4.4. --- Study 2 --- p.133
Chapter 4.4.1. --- Immunocytochemistry --- p.133
Chapter 4.5. --- Study 3 --- p.138
Chapter 4.5.1. --- Immunocytochemistry --- p.138
Chapter 4.5.2. --- RT-PCR --- p.141
Chapter 4.5.3. --- Real time RT-PCR --- p.145
Chapter 4.6. --- Endotoxin contamination in DBT --- p.149
Chapter 4.6.1. --- Effects of endotoxin on eNOS --- p.149
Chapter 4.6.2. --- Immunocytochemistry on immunostained endothelial cells --- p.151
Chapter 4.6.3. --- Effects of endotoxin on iNOS --- p.153
Chapter 4.6.4. --- Effect of endotoxin on NF-kB --- p.157
Chapter 4.6.5. --- Effects of endotoxin on ICAM-1 --- p.160
Chapter CHAPTER 5. --- DISCUSSION --- p.165
Chapter 5.1. --- DBT extraction and standardization of content --- p.165
Chapter 5.1.1. --- Optimal DBT extraction conditions --- p.165
Chapter 5.1.2. --- Evidence to support formulae usage --- p.166
Chapter 5.1.3. --- Limitation of the methodology used --- p.166
Chapter 5.2. --- Cell model development --- p.167
Chapter 5.2.1. --- Choice of DBT concentration range in the study --- p.167
Chapter 5.2.1.1. --- Choice of concentration range in consideration of endotoxin contamination --- p.167
Chapter 5.2.1.2. --- Choice of concentration range in consideration of DBT's cytotoxicity effects --- p.168
Chapter 5.2.1.3. --- Choice of concentration range in consideration of prevous studies --- p.168
Chapter 5.3. --- Study 1 --- p.169
Chapter 5.3.1. --- Action of DBT on eNOS expression --- p.169
Chapter 5.3.2. --- Action of DBT on iNOS expression --- p.170
Chapter 5.3.3. --- Action of DBT on Nitric oxide metabolite assay --- p.171
Chapter 5.3.3.1. --- Result interpretation with rejected hypothesis 2 --- p.171
Chapter 5.3.3.2. --- Assay limitations and improvements --- p.171
Chapter 5.4. --- Study 2 --- p.172
Chapter 5.4.1. --- Action of DBT on NF-kB expression --- p.172
Chapter 5.4.2. --- Assay limitations and improvements --- p.173
Chapter 5.5. --- Study 3 --- p.174
Chapter 5.5.1. --- Action of DBT on ICAM-1 expression --- p.174
Chapter 5.6. --- Endotoxin contamination in DBT --- p.175
Chapter 5.6.1. --- Action of endotoxin contamination in DBT on various markers --- p.175
Chapter 5.6:2. --- Experimental limitation --- p.176
Chapter 5.6.3. --- Endotoxin removal --- p.177
Chapter 5.6.3.1. --- Introduction --- p.177
Chapter 5.6.3.2. --- Endotoxin removal methodologies suitable for herbal use --- p.179
Chapter 5.7. --- Action of DBT on angiogenesis stimulation --- p.181
Chapter 5.7.1. --- Evidence for DBT's proangiogenic effects from various studies --- p.181
Chapter 5.7.2. --- Influence of endotoxin contamination on angiogenesis stimulation --- p.182
Chapter 5.7.3. --- Assay limitations and future developments --- p.183
Chapter CHAPTER 6. --- GENERAL DISCUSSION AND SUMMARY --- p.186
Chapter CHAPTER 7. --- REFERENCES --- p.191

碩士
大仁科技大學
製藥科技研究所
102
The research used single dose including Danggui Buxue Tang and Angelica: Astragalus (1:5), and Dendrobium candidum to assess the efficiency in vitro. The analysis is the reduction of α-diphenyl-β-picrylhydrazyl (DPPH) radical of the Dendrobium candidum Polysaccharides. Also, the concentration analysis have made, regarding to the active ingredients of Astragaloside contained in this prescription, ferulic acid and Dendrobium candidum Polysaccharides. We analyzed the active ingredients of ferulic acid ,which is the extract from water and alcohol in Danggui Buxue Tang. The results show that the content of ferulic acids were 7.25 ± 5.47 from water extract and 84.51 ± 11.31 g/g from alcohol extract, respectively. The content of ferulic acids from alcohol extract is higher. In the absorbance test of the Dendrobium candidum Polysaccharides, we had found that the most suitable concentration of the light absorption value is 300 g / mL in the OD (optical density) . To transfer the concentration of the light absorption value 300 g /mL of crude Dendrobium candidum Polysaccharides, according to a glucose standard curve values obtained by the phenol-sulfuric acid method, is equivalent to glucose 85 g/mL. Danggui Buxue Tang and Dendrobium candidum has show the capacity of reduction DPPH radical but the clear effect is weaker than the control group which given vitamin C. In the cytotoxicity assay (MTT assay), the research found that the combination of Danggui Buxue Tang extracts by alcohol and Dendrobium candidum Polysaccharides can significantly increase the survival rate of cells, and show protective effects in the rat intraocular pressure and retinal eye disease. The combination obviously has the better efficacy of blood sugar lowering test in hyperglycemic rats. Finally, we checked the data by biochemical and pathological observations results to find the considerable protective effect of kidney and liver in rats with the combination medicine treatment. These findings show reference to improve, even prevent, the hyperglycemia and retinopathy for clinical treatment.

RESUMO Título: A génese do tento no testemunho dos manuscritos P-Cug MM 48 e MM 242 (com uma edição crítica dos ricercari de Jacques Buus e das suas versões recompostas) A presente tese estuda a génese do tento e da fantasia para tecla na ilustração dos manuscritos P-Cug MM 48 e 242. Partindo das recomposições dos ricercari do Libro primo… (1547) de Jacques Buus constantes na parte inicial do MM 242 e tendo por base as suas cópias realizadas a partir do MM 48, esta tese tem como primeiro assunto central a análise dos processos de estudo e recomposição patentes nas referidas obras. Num segundo momento, é estabelecida a sua relação comparativa com a estrutura formal e estilística do tento e da fantasia para tecla em Portugal, por volta de meados do século XVI, no testemunho de algumas obras de António Carreira incluídas no MM 242. ABSTRACT Title: The origins of the tento in the testimony of manuscripts P-Cug MM 48 & MM 242 (with a critical edition of Jacques Buus´s ricercari and its recomposed versions) This thesis focuses on the origins of the keyboard tento and fantasia as exemplified by the two manuscripts P-Cug MM 48 & 242. Beginning with the recompositions of Jacques Buus´s ricercari from his Libro primo… (1547) that are in the initial part of MM 242 and having in mind that these were based upon the copies of the original versions included in MM 48, the first aim of this thesis is to analyze the study and recompositional processes that these works reveal. His second purpose concerns the comparative relation of Buus´s recompositions with the formal and stylistic structure of the mid-sixteenth century keyboard tento and fantasia in Portugal, as testified by some António Carreira´s work included in MM 242.