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Molecular authentication of epimedium species and the anti-osteoporosis effect of herba epimedii. / CUHK electronic theses & dissertations collection / Digital dissertation consortiumJanuary 2004 (has links)
Sun Ye. / "November 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 205-225) / 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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.
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Mechanistic study of herb-drug interactions between oseltamivir and TCM formulae. / Mechanistic study of herb-drug interactions between oseltamivir and traditional Chinese medicine formulaeJanuary 2010 (has links)
Wang, Xiaoan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 145-166). / Abstracts in English and Chinese. / Table of Contents --- p.I / Acknowledgements --- p.VI / Publications --- p.VII / Abstract (in English) --- p.VIII / Abstract (in Chinese) --- p.X / List of Figures --- p.XII / List of Tables --- p.XVI / List of Abbreviations --- p.XVII / Chapter Chapter One. --- Introduction --- p.1 / Chapter 1.1 --- Overview of oseltamivir --- p.1 / Chapter 1.1.1 --- General description of oseltamivir --- p.1 / Chapter 1.1.2 --- Pharmacological activities of oseltamivir --- p.3 / Chapter 1.1.3 --- Pharmacokinetics of oseltamivir --- p.3 / Chapter 1.1.3.1 --- Absorption of oseltamivir --- p.4 / Chapter 1.1.3.2 --- Distribution of oseltamivir --- p.5 / Chapter 1.1.3.3 --- Metabolism of oseltamivir --- p.6 / Chapter 1.1.3.4 --- Elimination of oseltamivir --- p.8 / Chapter 1.1.4 --- Side effects and toxicities of oseltamivir --- p.9 / Chapter 1.2 --- Overview of Chinese medicine formulae CMF1 (Yinqiaosan and Sangjuyin) --- p.9 / Chapter 1.2.1 --- Background and clinical use of CMF1 --- p.9 / Chapter 1.2.2 --- Quality control of CMF1 by manufacturer --- p.11 / Chapter 1.2.3 --- Major active components of CMF1 --- p.12 / Chapter 1.3 --- Previous studies on herb-drug interactions between O and CMF1 --- p.18 / Chapter 1.4 --- Rationale of the current study --- p.19 / Chapter 1.5 --- objectives --- p.19 / Chapter Chapter Two. --- Identification and quantification of major marker compounds in Yinqiaosan and Sangiuyin products --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials and methods --- p.23 / Chapter 2.2.1 --- Chemicals --- p.23 / Chapter 2.2.2 --- Instruments --- p.24 / Chapter 2.2.3 --- Chromatographic conditions --- p.24 / Chapter 2.2.4 --- Preparation of standard solutions --- p.25 / Chapter 2.2.5 --- Calibration curves --- p.26 / Chapter 2.2.6 --- Validation of the assay method --- p.26 / Chapter 2.2.7 --- Sample preparations for Yinqiaosan and Sangjuyin products --- p.27 / Chapter 2.2.7.1 --- Sample extraction from Yinqiaosan or Sangjuyin granules --- p.27 / Chapter 2.2.7.2 --- Sample extraction from Yinqiaosan or Sangjuyin tablets --- p.27 / Chapter 2.2.7.3 --- Sample extraction recoveries --- p.27 / Chapter 2.3 --- Results and discussions --- p.28 / Chapter 2.3.1 --- Chromatography --- p.28 / Chapter 2.3.2 --- Linearity and sensitivity --- p.33 / Chapter 2.3.3 --- Accuracy and precision --- p.33 / Chapter 2.3.4 --- Stability --- p.36 / Chapter 2.3.5 --- Contents of identified active components in commercial available Yinqiaosan or Sangjuyin products and CMF1 --- p.36 / Chapter 2.3.6 --- Sample extraction recovery --- p.40 / Chapter 2.4 --- Conclusion --- p.43 / Chapter Chapter Three. --- Effect of CMF1/CMF1 components on the metabolism of oseltamivir and related mechanistic studies --- p.44 / Chapter 3.1 --- Introduction --- p.44 / Chapter 3.2 --- Materials and methods --- p.47 / Chapter 3.2.1 --- Materials --- p.47 / Chapter 3.2.2 --- "Verification of metabolism of O in rat GI tract, plasma and liver microsome" --- p.48 / Chapter 3.2.3 --- Inhibition of metabolism of O by CMFl/CMFl components --- p.49 / Chapter 3.2.3.1 --- In vitro inhibition of metabolism of O in rat plasma --- p.49 / Chapter 3.2.3.2 --- In vitro inhibition of metabolism of O in rat liver microsome (RLM) --- p.49 / Chapter 3.2.4 --- Mechanistic study of enzyme inhibition of O in recombinant human Carboxylesterase 1 (hCE 1) --- p.50 / Chapter 3.2.5 --- Sample preparation and LC/MS/MS analysis --- p.50 / Chapter 3.2.6 --- Data analyses --- p.52 / Chapter 3.3 --- Results --- p.53 / Chapter 3.3.1 --- "Verification of metabolism of O in rat GI tract, plasma and liver microsome" --- p.53 / Chapter 3.3.2 --- Inhibition of metabolism of O by CMF1/CMF1 components --- p.53 / Chapter 3.3.2.1 --- Enzyme inhibition of metabolism of O by CMFl/CMF1 components in rat plasma --- p.53 / Chapter 3.3.2.2 --- Enzyme inhibition of metabolism of O by CMF1/CMF1 components in rat liver microsome (RLM) --- p.58 / Chapter 3.3.2.3 --- Selection of potent enzyme inhibitor from CMF1 --- p.60 / Chapter 3.3.4. --- Mechanistic study of enzyme inhibition of O in recombinant human Carboxylesterase 1 (hCE 1) --- p.61 / Chapter 3.4 --- Discussions --- p.63 / Chapter 3.5 --- Conclusion --- p.74 / Chapter Chapter Four. --- Effect of CMFl/CMFl components on the absorption of oseltamivir and related mechanistic studies --- p.75 / Chapter 4.1 --- Introduction --- p.75 / Chapter 4.2 --- Materials and methods --- p.79 / Chapter 4.2.1 --- Materials --- p.79 / Chapter 4.2.2 --- PAMPA permeation model --- p.80 / Chapter 4.2.2.1 --- Permeation of O and OC in PAMPA --- p.80 / Chapter 4.2.2.2 --- Sample preparation and LC/MS/MS analysis --- p.81 / Chapter 4.2.2.3 --- Data analysis --- p.81 / Chapter 4.2.3 --- Absorption of O in presence of CMF/CMFl components in Caco-2 and MDCK cell monolayer models --- p.82 / Chapter 4.2.3.1 --- Cell culture --- p.82 / Chapter 4.2.3.2 --- Preparation of loading solutions to the cell models --- p.83 / Chapter 4.2.3.3 --- Stability of O in transport buffer --- p.84 / Chapter 4.2.3.4 --- Cytotoxicity tests of O and CMFl/CMFl components --- p.84 / Chapter 4.2.3.5 --- Transport study in Caco-2 and MDCK monolayer model --- p.85 / Chapter 4.2.3.6 --- Sample preparation and LC/MS/MS analysis --- p.86 / Chapter 4.2.3.7 --- Data analysis --- p.87 / Chapter 4.2.4 --- Absorption of O in presence of CMF 1 in rat in situ single pass intestinal perfusion model --- p.88 / Chapter 4.2.4.1 --- Preparation of perfusion solutions --- p.88 / Chapter 4.2.4.2 --- Stabilities of O and arctigenin in perfusate --- p.88 / Chapter 4.2.4.3 --- Rat in situ single pass intestinal perfusion of O in presence and absence of CMFl and relevant inhibitors --- p.89 / Chapter 4.2.4.4 --- Sample preparation and LC/MS/MS analysis --- p.90 / Chapter 4.2.4.5 --- Data analysis --- p.90 / Chapter 4.3 --- Resul ts --- p.91 / Chapter 4.3.1 --- Permeation of O and OC in PAMPA --- p.91 / Chapter 4.3.2 --- Absorption of O in presence of CMF/CMF1 components in Caco-2 and MDCK cell monolayer models --- p.92 / Chapter 4.3.2.1 --- Stabilities of O in transport buffer --- p.92 / Chapter 4.3.2.2 --- Cytotoxicity tests of O and CMF1/CMF1 components in transport buffer --- p.93 / Chapter 4.3.2.3 --- Proof of O as a substrate of P-gp by Caco-2 cell model --- p.95 / Chapter 4.3.2.4 --- Effect of CMF 1 on the absorption transport of o in Caco-2 cell mode --- p.98 / Chapter 4.3.2.5 --- Effect of CMF1 components on the absorption transport of o in Caco-2 cell model --- p.102 / Chapter 4.3.2.6 --- Effect of arctigenin on bi-directional transport of o in Caco- 2 cell model --- p.106 / Chapter 4.3.2.7 --- Proof of O as a substrate of P-gp by MDCK transfected cell lines --- p.108 / Chapter 4.3.2.8 --- Bi-directional transport of O in MDCK-MDR1 cell model --- p.111 / Chapter 4.3.2.9 --- Effect of CMF 1 on the absorption transport of O in MDCK-MDR1 cell model --- p.112 / Chapter 4.3.3 --- Absorption of O in presence of CMF1 in rat in situ single pass intestinal perfusion model --- p.113 / Chapter 4.3.3.1 --- Stabilities of O and arctigenin in the perfusion buffer --- p.113 / Chapter 4.3.3.2 --- Intestinal absorption of O in presence and absence of CMF1 in rat in situ intestinal perfusion model --- p.114 / Chapter 4.4 --- Discussions --- p.116 / Chapter 4.5 --- Conclusion --- p.124 / Chapter Chapter Five. --- Preliminary evaluation of antiviral activity of CMFl/CMFl components --- p.125 / Chapter 5.1 --- Introduction --- p.125 / Chapter 5.2 --- Materials and methods --- p.128 / Chapter 5.2.1 --- Materials and animals --- p.128 / Chapter 5.2.2 --- Animal treatment --- p.129 / Chapter 5.2.3 --- Plasma sample collection and preparation --- p.130 / Chapter 5.2.4 --- Evaluation of antiviral activities of CMFl/ CMFl components --- p.130 / Chapter 5.2.4.1 --- Plaque reduction assay --- p.131 / Chapter 5.2.4.2 --- Optimization of plasma sample dilution ratio --- p.131 / Chapter 5.2.5 --- Data analyses --- p.133 / Chapter 5.3 --- Results and discussions --- p.135 / Chapter 5.3.1 --- Ex vivo evaluation of antiviral activity of CMF1 --- p.135 / Chapter 5.3.2 --- In vitro evaluation of antiviral activity of CMF1 major marker compounds --- p.139 / Chapter 5.4 --- Conclusion --- p.141 / Chapter Chapter Six. --- Overall conclusion --- p.142 / References --- p.145
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Efeito da eletroacupuntura na qualidade de vida de pacientes com esclerose múltipla forma recorrente-remitente / Impact of electroacupuncture on quality of life for patients with relapsing-remitting multiple sclerosisQuispe Cabanillas, Juan Guzman, 1974- 30 January 2006 (has links)
Orientadores: Wanderley Dias da Silveira, Leonilda Maria Barbosa dos Santos / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-20T04:05:25Z (GMT). No. of bitstreams: 1
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Previous issue date: 2012 / Resumo: A Acupuntura é uma das mais antigas formas de tratamento que tem sobrevivido e evoluido não só no Extremo Oriente (China), mas em todo o mundo. A técnica consiste na inserção de agulhas em vários níveis de profundidade da pele em pontos específicos chamados de "Acupontos", podendo ser estimuladas manualmente ou com uma corrente de baixa tensão elétrica (Eletroacupuntura). Faz parte ainda de um conjunto de técnicas com fins terapêuticos da Medicina Tradicional Chinesa, sendo, atualmente, praticada como terapia primária e adjuvante para uma variedade de doenças e condições crônicas. De outro lado a Esclerose Múltipla é uma doença inflamatória crônica do sistema nervoso central, caracterizada pela infiltração de células do sistema imune neste compartimento, com destruição de mielina e perda de oligodendrócitos, é a mais comum das doenças auto-imunes do sistema nervoso central. Trata-se de uma doença que acomete principalmente adultos jovens, sendo mais frequente nas mulheres, causando múltiplos sinais e sintomas de disfunção neurológica. Aproximadamente 85% dos pacientes apresentam a forma recorrente - remitente (surto/remissão), que é caraterizada por apresentar manifestações sintomatológicas de disfunção neurológica (exacerbação, relapso ou ataque) com períodos de estabilização ou melhora (remissão). Terapias imunomoduladoras modernas vêm sendo empregadas no tratamento da Esclerose Múltipla, no entanto, não aliviam muitos dos sintomas, como dor e depressão, levando a uma piora na qualidade de vida. Pacientes, assim, procuram tratamentos alternativos, como a acupuntura, embora os benefícios desses tratamentos não tenham sido objetivamente avaliados. O emprego do modelo experimental, a Encefalomielite Autoimune (alérgica) Experimental, tem trazido grandes contribuições para a compreensão dos mecanismos envolvidos e o esclarecimento da patogênese de doenças autoimunes, tais como a Esclerose Múltipla. Este estudo foi, portanto, concebido para avaliar o efeito da eletroacupuntura sobre a qualidade de vida de pacientes com Esclerose Múltipla tratados com imunomoduladores e na modulação da resposta imune na encefalomielite autoimune experimental. No final do estudo foi observado que a Eletroacupuntura promove a melhora em vários aspectos da qualidade de vida dos pacientes com Esclerose Múltipla, especialmente a dor, e na Encefalomielite Autoimune (alérgica) Experimental foi observado que existe uma modificação da resposta imunológica. Além disso, os resultados também sugerem que o uso rotineiro de um questionário de autoavaliação da qualidade da vida deve ser incluído nas avaliações clínicas regulares dos pacientes / Abstract: Acupuncture is one of the oldest forms of treatment that has survived and evolved not only in the Far East (China), but around the world. The technique involves inserting needles at various levels of depth of the skin at specific points called "Acupoints" and can be stimulated manually or with a low voltage electrical current (electroacupuncture). It is part of a set of therapeutic techniques the traditional chinese medicine and is currently practiced as primary therapy and adjuvant for a variety of diseases and chronic conditions. On the other hand the MS is a chronic inflammatory disease of the central nervous system characterized by infiltration of immune cells in this compartment, with destruction of myelin and oligodendrocytes loss. It is the most common of the autoimmune diseases of the central nervous system and is a disease that mostly affects young adults, being more common in women, causing multiple signs and symptoms of neurological dysfunction. Approximately 85% of patients present form recurrent-remitting MS (relapsing/remitting), which presenting symptom of neurological events (exacerbation, relapse or attack) with periods of stabilization or improvement (remission). Modern immunomodulatory therapies have been employed in the treatment of Multiple Sclerosis, however, do not alleviate many of the symptoms such as pain and depression, leading to deterioration in quality of life. Patients, therefore, seek alternative treatments such as acupuncture, although the benefits of these treatments have not been objectively evaluated. The use of model experimental autoimmune encephalitis has brought large contributions to the understanding of mechanisms and understanding the pathogenesis of autoimmune diseases such as MS. This study was therefore designed to evaluate the effect of electro on the quality of life of patients with multiple sclerosis treated with immunomodulators and modulation of immune response in experimental autoimmune encephalomyelitis. At the end of the study it was observed that electroacupuncture improved in various aspects of quality of life of patients with multiple sclerosis, especially pain, and it was observed that electroacupuncture modifies the immune response in the Autoimmune Encephalomyelitis Experimental. Moreover, the results also suggest that the routine use of a self-assessment of quality of life should be included in regular clinical evaluations of patients / Doutorado / Farmacologia / Mestre em Farmacologia
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Cardiovascular effects of Rhizoma chuanxiong and its active constituents. / CUHK electronic theses & dissertations collectionJanuary 2005 (has links)
In a mouse model of pulmonary thromboembolism induced by a collagen-adrenaline mixture, the SFE extract and ligustilide reduced the paralysis-death ratio, and the anti-thrombotic response of senkyunolide A was more pronounced. The effect of BDPH was not significant. Neither the SFE extract nor the three phthalides prolonged bleeding time in tail-transected mice. / In a rat myocardial ischemia-reperfusion model involving coronary artery ligation, 7-day pre-treatment with the SFE extract and ligustilide reduced ventricular arrhythmias in isolated hearts. BDPH and senkyunolide A were without significant effects. / In rat platelet-rich plasma, platelet aggregation induced by collagen and U46619 but not by adenosine diphosphate was inhibited by the SFE extract. Ligustilide inhibited the responses of all three agonists, while BDPH and senkyunolide A inhibited the collagen response only. / Raw Rhizoma Chuanxiong herb and its crude extract as obtained by supercritical fluid extraction (SFE) comprised mainly phthalides. The SFE extract and three representative phthalides, butylidenephthalide (BDPH), ligustilide and senkyunolide A, were studied on vasorelaxation, myocardial ischemia, platelet aggregation and thrombosis. The mechanisms underlying BDPH-mediated vasorelaxation were also explored. / Rhizoma Chuanxiong, the dried rhizome of Ligusticum chuanxiong Hort., is a common traditional Chinese medicine used for the treatment of cardiovascular diseases. Surprisingly, the scientific basis of its action is poorly understood. The current study aims to establish the pharmacological basis of the cardiovascular effects of Rhizoma Chuanxiong and its active constituents by examining their effects in several cardiovascular domains. / The current study demonstrated various cardiovascular actions of Rhizoma Chuanxiong, and thereby established the pharmacological basis of the effects of the herb. Phthalides, in particular BDPH, ligustilide and senkyunolide A, were important contributors to such actions. Future investigation of the SFE extract and/or individual phthalides related to the progression from in vitro and in vivo effectiveness to clinical efficacy is much anticipated. / The SFE extract, BDPH, ligustilide and senkyunolide A produced vasorelaxation on isolated preparations of rat aorta, rat saphenous vein and pig coronary artery. BDPH-mediated relaxation appeared to involve both extracellular Ca 2+-dependent (L-type voltage-operated, receptor-operated and store-operated Ca2+ channels) and independent (NO modulation, Ca2+ release from internal stores and Ca2+ desensitization) mechanisms. BDPH was also observed to augment relaxation induced by sodium nitroprusside and forskolin through mechanisms that remain undefined. / Chan Sun Kin Sunny. / "July 2005." / Advisers: G. Lin; R. L. Jones. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1575. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 190-209). / 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 English and Chinese. / School code: 1307.
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Anticancer activity and mechanistic study of a series of platinum complexes integrating demethylcantharidin with isomers of 1,2-diaminocyclohexane. / CUHK electronic theses & dissertations collectionJanuary 2006 (has links)
Aim. The aim of this study was to synthesize and characterize novel analogues of [DACH-Pt-DMC] by using different stereoisomers of DACH; and to investigate any differences in in vitro activity of these complexes in human hepatocellular carcinoma (HCC), colorectal carcinoma (CRC) cell lines and acquired cisplatin or oxaliplatin resistant sub-lines, and to compare that of oxaliplatin and other established Pt-based anticancer agents. Mechanistic roles of DACH-Pt- and DMC components of the TCM-Pt complexes on affecting HCT 116 human CRC cell line were investigated by flow cytometry, COMET assay and cDNA microarray analysis. / Background. Demethylcantharidin (DMC), a modified component of the traditional Chinese medicine (TCM), integrated with a platinum (Pt) moiety created a series of TCM-Pt complexes [Pt(C8H8O 5)(NH2R)2] 1-5 which demonstrated superior antitumor activity and circumvention of cisplatin resistance in vitro. Compound 5, derived from the 1,2-diaminocyclohexane (DACH) ligand (where R=trans-C6H10) had the most potent antitumor activity and closest structural resemblance to oxaliplatin (R,R-DACH-Pt complex) which is the first Pt-based anticancer drug to demonstrate convincing clinical activity against colorectal cancer and has a mechanism of action and resistance that is clearly different from that of cisplatin and carboplatin. / Conclusion. This study is the first to examine the mechanism of anticancer activity of new complexes that integrate DMC with different isomers of DACH. It has shown that both DACH-Pt- and DMC components contribute significantly to the compounds' potent anticancer activity, but likely with different mechanisms of action. The DACH-Pt- component appears to dictate the cell cycle distribution, whereas the DMC component appears to enhance cytotoxicity by inducing more DNA damage in HCT 116 colorectal cancer cells. / Methods. DMC was reacted with appropriate DACH-Pt-(NO3) 2 intermediates, which were prepared from treatment of K2PtCl 4 with stereoisomeric DACH (RR-, SS- & cis-), followed by reaction with silver nitrate. Proton NMR, high-resolution MS, polarimetry and circular dichroism (CD) spectroscopy were used to characterize their chemical structures and optical activities. In vitro antitumor activity (IC50 of 72hr drug exposure time) were assessed by a standard MTT assay. Cell cycle analysis by flow cytometry was determined at 0, 6, 12, 18, 24, 48 and 72 h after drug treatment (cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or trans-DACH-Pt-DMC analogues) at IC50 and 5 x IC50 concentrations with three to four replicates. Comet assay was performed with a fluorescent microscope and used to examine DNA damage after drug treatments (50muM of cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or R,R-DACH-Pt-DMC) for 3hr. cDNA microarray was performed on Affymetrix Human Genome U133A Set and used to analyze gene expression profiles in HCT 116 exposed to trans-(+/-)-DACH-Pt-DMC or oxaliplatin at their IC50 for 72hr. / Results. The in vitro results showed that the trans-analogues were consistently the most potent amongst all the compounds tested in both HCC and CRC cell lines: the trans-(+)(1R,2R)-DACH-Pt-DMC complex, in particular, was the most effective stereoisomer. All of the stereoisomeric DACH-Pt-DMC complexes and oxaliplatin were apparently able to circumvent cisplatin resistance in Huh-7 and SK-Hep1 sub-lines, but cross resistant with oxaliplatin in HCT 116 oxaliplatin resistant sub-line. Flow cytometric analysis revealed the novel trans-DACH-Pt-DMC analogues and oxaliplatin behaved similarly: that is, the compounds at 5 x IC50 concentrations all caused a significant decrease in the S-phase population within 18h and at the same time induced G2/M arrest, and without obvious sub-G 1 phase accumulation, but distinct from that of cisplatin, carboplatin or DMC. Comet assay showed that trans-(+)-(1R,2 R)-DACH-Pt-DMC caused the most significant DNA damage at an equivalent molar concentration. Microarray analysis suggested that the mechanistic role of the DMC ligand can induce the cell cycle to accelerate from the G 1 to S-phase and cause M-phase arrest. / Yu Chun Wing. / "July 2006." / Advisers: Yee-ping Ho; Chik Fun Steve Au-Yeung. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1586. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 191-232). / 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 English and Chinese. / School code: 1307.
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Development and in-vitro evaluation of peroral and buccoadhesive formulations for biologically active crude oil extracted from Ligusticum chuanxiong, a traditional Chinese medicine. / CUHK electronic theses & dissertations collectionJanuary 2005 (has links)
differential scanning calorimetric profile and the generation of much less intense and broader peaks in the powder X-ray diffraction pattern compared to beta-CD. FTIR analysis revealed significant physical interactions between CX oil and beta-CD in the granules, possibly due to complexation. Results from phase solubility measurements and proton nuclear magnetic resonance ( 1H-NMR) analysis of pure 3-butylidenephthalide (3-BDPH), a representative CX component, lend some support for the formation of a 1:1 stoichiometric inclusion complex between 3-BDPH and beta-CD. / Rhizoma chuanxiong (CX), the dried rhizome of Ligusticum Chuangxiong Hort. (Umbelliferae), has been extensively used in mainland China as a traditional herbal medicine for treating cardio-/cerebrovascular diseases and gynecological disorders. However, the active components in CX, which are predominantly essential oils, generally exhibit poor stability (mostly photo-oxidation), high volatility, low aqueous solubility, and extensive gut/hepatic metabolism, all of which can significantly reduce their oral bioavailability and therapeutic efficacy. The present project has investigated the feasibility of utilizing three formulation approaches to circumvent the aforementioned problems associated with the peroral delivery of CX (as crude oil mixture or individual components). / The first approach involved inclusion of CX oil in beta-cyclodextrin (beta-CD) as solid granules using a coprecipitation method optimized with the aid of an orthogonal study design. The resulting CX oil granules were colorless and odorless with a median particle size of 11.38mum; were stable to heat, light and moisture, and readily soluble in simulated gastric and intestinal fluids. The granules were largely amorphous, as evidenced by an absence of the melting endotherm for beta-CD in the formulation could be largely explicated by the complexation behavior and hydration properties of the two polymers blended in different weight percentages, as substantiated by turbidity measurement, viscosity determination and FTIR analysis of the pure polymer mixtures as well as swelling measurement of the formulated tablets. The sustained release behavior of 3-BDPH from the tablet was dependent on the relative proportion of the two polymers present, and could be similarly explained by the changes in hydration and complexation behavior of the polymers during the penetration of aqueous fluid into the tablet matrix. / The second approach involved incorporation of CX oil into surfactant micelles and liquid crystals as a self-emulsifying drug delivery system (SEDDS). An optimal formulation was developed through a judicial choice of excipients (lipids and surfactants/cosurfactant) and their proper combination in the correct proportions, as determined by the spontaneity of the emulsification process and the change in emulsion droplet size. The formulation was readily dispersible in water upon mild agitation, free from unpleasant odor, and stable in soft gelatin capsules for a storage period of at least 12 months under ambient condition. The optimal utilization of the lipid and surfactant blends in defined proportions in the formulation was further substantiated by interfacial tension determination and equilibrium phase analysis. / The third approach involved formulation of 3-BDPH (or crude CX oil) into a sustained-release buccoadhesive tablet, based on a systematic evaluation of the adhesive properties of two polymers (Carbopol 974P and hydroxypropyl methylcellulose K4M) used in the formulation. The adhesive properties of the formulation could be largely explicated by the complexation behavior and hydration properties of the two polymers blended in different weight percentages, as substantiated by turbidity measurement, viscosity determination and FTIR analysis of the pure polymer mixtures as well as swelling measurement of the formulated tablets. The sustained release behavior of 3-BDPH from the tablet was dependent on the relative proportion of the two polymers present, and could be similarly explained by the changes in hydration and complexation behavior of the polymers during the penetration of aqueous fluid into the tablet matrix. / Gao Yuan. / "April 2005." / Adviser: Albert H. L. Chow. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1585. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 193-223). / 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 English and Chinese. / School code: 1307.
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HPLC-MS analysis of radix astragali, cortex phellodendri, rhizoma coptidis and sanhuang xiexin decoction /cTsai, Sam Hip. / CUHK electronic theses & dissertations collectionJanuary 2007 (has links)
A method is presented for the simultaneous identification of nine compounds in samples of A. membranaceus and A. membranaceus var. mongholicus. Compounds identified in the extracts of the two plants included glycosides, saponins and flavonoids. They are identified as Calycosin-7-O-beta-D-glucoside (C1), Ononin (C2), (6aR,11aR)-3-hydroxy- 9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (C3), (3R)-7,2'-dihydr- oxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside (C4), Calycosin (C6), Astragaloside IV (C5 ), Formononetin (C7), (6aR,11a R)-3-hydroxy-9,10-di-methoxypterocarpan (C8), and Isomucronulatol (C9). / An HPLC-DAD-MS method is proposed for the differentiation of Rhizoma Coptidis and Cortex Phellodendri samples. This method can also be used to identify two common species of Rhizome Coptidis, i.e., C. chinensis and C. deltoidea, and two species of Cortex Phellodendri, i.e., P. chinensis and P. amurense. From the experiment results, there are thirteen, twelve and seven common components found in samples Rhizoma Coptidis, P. amurense and P. chinensis, respectively. Nine compounds in Rhizoma Coptidis were identified to be alkaloids. The common components in Cortex Phellodendri included four alkaloids and two lactones, i.e., obaculacotone and obacunone, present in all samples of P. amurense. / High Performance Liquid Chromatography-Atmospheric Pressure Chemical Ionization Mass-Spectrometry has been applied to the analysis and standardization of Chinese Herbal Medicines. The applications included quantitative study of Astragaloside in Radix Astragali, investigation on the chromatographic fingerprint of Radix Astragali, differentiation of Cortex Phellodendri and Rhizoma Coptidis, and identification of constituents in Sanhuang Xiexin Decoction. / In the quantitative study of Astragaloside, an Multiple Reaction Monitoring scan mode was used. The linearity between 2 and 500 mg/L is 0.9996. The precision of injection and reproducibility of method is 1.72% and 3.27% respectively. A total of 20 samples from local market and mainland China were analyzed and the results are comparable to those obtained from HPLC-ELSD analysis. / The present study also proposed a HPLC separation and online identification for the 15 constituents in a composite Chinese herbal formula, the Sanhuang Xiexin Decoction. It provided a possible starting point to evaluate related herbal preparations containing Rhizoma Coptidis, Radix Scutellariae and Rhizoma Rhei. Thirteen constituents in the decoction were identified, including five major alkaloids from Rhizoma Coptidis, five anthraquinones from Rhizoma Rhei and two favonoids and one glycoside from Radix Scutellariae. / "November 2007." / Adviser: Chi Chun Tao. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4726. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 177-200). / 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 English and Chinese. / School code: 1307.
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Immunomodulatory effects and toxicity of mimosa pudica, the sensitive plant.January 1993 (has links)
by Cheng Yuk Kwan, Anna. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 104-112). / Acknowledgements / Table of Contents --- p.i / Abbreviations --- p.iv / Abstract --- p.vi / List of figures --- p.ix / List of tables --- p.xi / Chapter Chapter One: --- Introduction / Chapter 1.1 --- Objective and scope of the project --- p.1 / Chapter 1.2 --- Literature review of Mimosa pudica / Chapter 1.2.1 --- Morphology of Mimosa pudica --- p.3 / Chapter 1.2.2 --- Chemistry of Mimosa pudica --- p.5 / Chapter 1.2.3 --- Uses in traditional medicine --- p.5 / Chapter 1.2.4 --- Clinical and pharmacological studies of Mimosa pudica --- p.6 / Chapter 1.2.5 --- Toxicology of Mimosa pudica --- p.8 / Chapter 1.2.6 --- Characteristics and toxicology of mimosine --- p.9 / Chapter 1.3 --- Immunomodulation / Chapter 1.3.1 --- Overview of the immune system --- p.11 / Chapter 1.3.2 --- Strategies on the study of immunomodulation of Mimosa pudica --- p.13 / Chapter 1.4 --- Toxicology / Chapter 1.4.1 --- Principles of the toxicological assays / Chapter 1.4.1.1 --- LD50 --- p.17 / Chapter 1.4.1.2 --- Enzyme assays --- p.18 / Chapter 1.4.1.3 --- Subacute toxicity test --- p.24 / Chapter 1.4.1.4 --- Reproductive toxicity test --- p.25 / Chapter Chapter Two: --- Materials and methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Mimosa pudica --- p.27 / Chapter 2.1.2 --- Animals --- p.27 / Chapter 2.1.3 --- Chemicals --- p.28 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Extraction of Mimosa pudica --- p.32 / Chapter 2.2.2 --- Assays for the immunomodulatory effects of Mimosa pudica / Chapter 2.2.2.1 --- Cell preparation / Chapter a) --- Splenocytes --- p.35 / Chapter b) --- Thymocytes --- p.35 / Chapter c) --- Macrophages --- p.36 / Chapter 2.2.2.2 --- Splenocyte proliferation --- p.37 / Chapter 2.2.2.3 --- Thymocyte proliferation --- p.38 / Chapter 2.2.2.4 --- Phagocytic activity of macrophages --- p.39 / Chapter 2.2.2.5 --- Release of IL-1 by macrophages --- p.40 / Chapter 2.2.2.6 --- Plaque forming cells --- p.41 / Chapter 2.2.2.7 --- Restoration on splenocyte blastogenesis of old mice --- p.42 / Chapter 2.2.3 --- Assays for the toxicity of Mimosa pudica / Chapter 2.2.3.1 --- LD50 --- p.43 / Chapter 2.2.3.2 --- Enzyme assays --- p.43 / Chapter 2.2.3.3 --- Subacute toxicity --- p.43 / Chapter 2.2.3.4 --- Reproductive toxicity --- p.44 / Chapter 2.2.4 --- Statistical analysis --- p.44 / Chapter Chapter Three: --- Results / Chapter 3.1 --- Immunomodulatory effects of Mimosa pudica / Chapter 3.1.1 --- In vitro study on the lymphocyte proliferation / Chapter 3.1.1.1 --- Splenocyte proliferation --- p.45 / Chapter 3.1.1.2 --- Thymocyte proliferation --- p.50 / Chapter 3.1.2 --- In vivo study on the lymphocyte proliferation --- p.53 / Chapter 3.1.3 --- Phagocytic activity of macrophages --- p.58 / Chapter 3.1.4 --- Release of IL-1 by macrophages --- p.64 / Chapter 3.1.5 --- Plaque forming cells --- p.67 / Chapter 3.1.6 --- Restoration on splenocyte blastogenesis of old mice --- p.69 / Chapter 3.2 --- Toxicity of Mimosa pudica / Chapter 3.2.1 --- LD50 --- p.72 / Chapter 3.2.2 --- Enzyme assays --- p.75 / Chapter 3.2.3 --- Subacute toxicity --- p.80 / Chapter 3.2.4 --- Reproductive toxicity --- p.85 / Chapter Chapter Four: --- General discussion on the immunomodulatory effects and toxicity of Mimosa pudica / Chapter 4.1 --- Immunomodulatory effects of Mimosa pudica --- p.88 / Chapter 4.2 --- Toxicity of Mimosa pudica --- p.95 / Chapter Chapter Five: --- Concluding remarks --- p.99 / References --- p.104 / Appendix --- p.113
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Effects of hepato-protective herbal medicines on gene expression in rat hepatocytes and hepatoma cells.January 2002 (has links)
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
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Aryl hydrocarbon receptor-mediated transcription and CYP1 class gene expression: could it be a possible mode of action of traditional chinese medicine in the management of breast carcinoma?. / 芳香烴受體介導的轉錄與CYP一組基因表達: 會不會是中藥治理乳癌的一個可能作用方法? / Fang xiang jing shou ti jie dao de zhuan lu yu CYP yi zu ji yin biao da: hui bu hui shi Zhong yao zhi li ru ai de yi ge ke neng zuo yong fang fa?January 2009 (has links)
Cheung, Tsz Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 97-116). / Abstracts in English and Chinese. / Thesis/Assessment Committee Members --- p.ii / Declaration for Plagiarism and Copyright --- p.iii / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.viii / Table of Contents --- p.ix / List of Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xvi / Chapter CHAPTER TWO: --- Introduction / Chapter 1.1 --- Background Information / Chapter 1.1.1 --- Breast Cancer --- p.1 / Chapter 1.1.2 --- General Statistics of Breast Cancer Worldwide and in Hong Kong --- p.1 / Chapter 1.1.3 --- Risk Factors for Breast Cancer --- p.2 / Chapter 1.1.4 --- Breast Cancer Treatment and Side Effects --- p.2 / Chapter 1.1.5 --- Types of Breast Cancer --- p.3 / Chapter 1.2 --- Estrogen and Estrogen Receptor / Chapter 1.2.1 --- Estrogen --- p.4 / Chapter 1.2.2 --- Estrogen Receptor --- p.5 / Chapter 1.2.3 --- Estrogen Receptor mediated Gene Transcription --- p.5 / Chapter 1.2.4 --- Estrogen Receptor Alpha and Estrogen Receptor Beta --- p.6 / Chapter 1.2.5 --- Estrogen Receptor Positive Breast Cancer and Treatment --- p.7 / Chapter 1.3 --- Estrogen metabolism and Cytochrome P450 family 1 (CYP1) members / Chapter 1.3.1 --- Estrogen Metabolism in Human --- p.9 / Chapter 1.3.2 --- CYP1A1 and CYP1B1 --- p.9 / Chapter 1.3.3 --- Estrogen Metabolism in Breast --- p.10 / Chapter 1.3.4 --- Carcinogenesis of Estrogens and Estrogen Metabolites --- p.13 / Chapter 1.3.5 --- The Importance of CYP1B1 in Carcinogenesis --- p.15 / Chapter 1.4 --- Aryl Hydrocarbon Receptor / Chapter 1.4.1 --- General Information of Aryl Hydrocarbon Receptor --- p.16 / Chapter 1.4.2 --- Signaling/Regulation Pathways of Aryl Hydrocarbon Receptor --- p.17 / Chapter 1.4.3 --- Crosstalk with Estrogen Receptor --- p.17 / Chapter 1.5 --- Introduction of Herba Scutellaria Barbata and its active ingredient Pheophorbide a --- p.19 / Chapter 1.6 --- Hyposthesis and Objectives --- p.21 / Chapter CHAPTER TWO: --- Direct Cytotoxic/Cytostatic Effect of Pheophorbide a / Chapter 2.1 --- Backgrounds --- p.22 / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Chemicals --- p.24 / Chapter 2.2.2 --- Cell Lines --- p.26 / Chapter 2.2.3 --- "Cell Culture Mediums, Buffers and Consumables" / Chapter 2.2.3.1 --- Roswell Park Memorial Institute Tissue Culture Medium1640 (RPMI1640) --- p.26 / Chapter 2.2.3.2 --- RPMI 1640 (Phenol Red-free) --- p.26 / Chapter 2.2.3.3 --- Serum supplement - Fetal Bovine Serum (FBS) --- p.27 / Chapter 2.2.3.4 --- Serum supplement - Charcoal/Dextran Stripped FBS --- p.27 / Chapter 2.2.3.5 --- Antibiotics - Penicillin-Streptomycin (P/S) --- p.27 / Chapter 2.2.3.6 --- Trypsin (0.25%) with EDTA --- p.27 / Chapter 2.2.3.7 --- Trypsin (2.5%) (Phenol Red-free) with EDTA --- p.28 / Chapter 2.2.3.8 --- Dulbeccóةs Phosphate-Buffered Saline (D-PBS) --- p.28 / Chapter 2.2.3.9 --- Tissue Culture Flasks and Multi-well Plate --- p.28 / Chapter 2.2.3.10 --- Trypan Blue Solution --- p.29 / Chapter 2.2.4 --- Reagents for Direct Cytotoxity Test / Chapter 2.2.4.1 --- MTT Assay --- p.29 / Chapter 2.2.4.2 --- Tritiated Thymidine Incorporation Assay --- p.29 / Chapter 2.3 --- Methods / Chapter 2.3.1 --- Cell Culture --- p.30 / Chapter 2.3.2 --- Direct Cytotoxicity/Cytostatic Test / Chapter 2.3.2.1 --- MTT Assay --- p.31 / Chapter 2.3.2.2 --- Tritiated Thymidine Incorporation Assay --- p.32 / Chapter 2.3.3 --- Statistical Analysis --- p.32 / Chapter 2.4 --- Results / Chapter 2.4.1 --- The Cytotoxic Effect of Pheophorbide a --- p.34 / Chapter 2.4.2 --- The Combine Effect of Pheophorbide a with 17-β Estradiol and Tamoxifen Citrate --- p.34 / Chapter 2.5 --- Discussions --- p.48 / Chapter CHAPTER THREE: --- Mechanistic Study of Pheophorbide a / Chapter 3.1 --- Backgrounds --- p.53 / Chapter 3.2 --- Materials / Chapter 3.2.1 --- Real time PCR / Chapter 3.2.1.1 --- General Chemicals and Equipments --- p.54 / Chapter 3.2.1.2 --- RNA isolation --- p.55 / Chapter 3.2.1.3 --- Reverse Transcription --- p.55 / Chapter 3.2.1.4 --- Real Time PCR --- p.56 / Chapter 3.2.2 --- Western Blotting / Chapter 3.2.2.1 --- Microsome Isolation --- p.58 / Chapter 3.2.2.2 --- Measurement of Protein Concentration --- p.58 / Chapter 3.2.2.3 --- Western Blotting --- p.58 / Chapter 3.2.3 --- Estrogen Metabolism Assay / Chapter 3.2.3.1 --- Chemicals --- p.59 / Chapter 3.2.3.2 --- Estrogen Metabolites Extraction --- p.60 / Chapter 3.2.3.3 --- Liquid Chromatography/Mass Spectrometry --- p.60 / Chapter 3.3 --- Methods / Chapter 3.3.1 --- Real time PCR / Chapter 3.3.1.1 --- Cell Culture --- p.61 / Chapter 3.3.1.2 --- RNA Isolation and Reverse Transcription --- p.61 / Chapter 3.3.1.3 --- Real Time PCR --- p.62 / Chapter 3.3.2 --- Western Blotting / Chapter 3.3.2.1 --- Cell Culture --- p.63 / Chapter 3.3.2.2 --- Microsome Isolation --- p.63 / Chapter 3.3.2.3 --- Measurement of Protein Concentration --- p.64 / Chapter 3.3.2.4 --- Western Blotting --- p.64 / Chapter 3.3.3 --- Estrogen Metabolism Assay / Chapter 3.3.3.1 --- Preparation of Calibration Standard --- p.65 / Chapter 3.3.3.2 --- Cell Culture --- p.66 / Chapter 3.3.3.3 --- Estrogen Metabolites Extraction --- p.66 / Chapter 3.3.3.4 --- Liquid Chromatography/Mass Spectrometry --- p.67 / Chapter 3.3.4 --- Statistical Analysis --- p.68 / Chapter 3.4 --- Results --- p.69 / Chapter 3.5 --- Discussions --- p.80 / Chapter CHAPTER FOUR: --- Overall Conclusion and Future Directions / Chapter 4.1 --- Significance of the Study --- p.87 / Chapter 4.2 --- Overall Conclusion --- p.87 / Chapter 4.3 --- Limitation and Difficulties of the Study --- p.89 / Chapter 4.4 --- Future Directions --- p.89 / Appendices / "Appendix I The Melting Curve of real time PCR for β-actin, CYP1A1 and CYP1B1" --- p.92 / Appendix II The Calibration Curve of BSA for Protein Concentration Measurement --- p.93 / Appendix III The Representative Peak of Estradiol Metabolite Standards with corresponding Retention Time --- p.94 / Appendix IV The Calibration Curve of Different Estrogen Metabolites for LC/MS --- p.95 / Appendix V The Accuracy and Precision of Quality Control of Estradiol Metabolites --- p.96 / Bibliography --- p.97
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