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Anti-HBV effects of three phyllanthus species and purification of its active component.

Lam Kit. / Thesis submitted in: July 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 141-153). / Abstracts in English and Chinese. / Acknowledgment --- p.I / Table of Content --- p.II / List of Tables --- p.VII / List of Figures --- p.IX / Abbreviations --- p.XIV / Abstract --- p.XVI / 論文摘要 --- p.XIX / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Hepatitis B --- p.1 / Chapter 1.1.1 --- Brief Introduction of HBV --- p.1 / Chapter 1.1.2 --- History of Hepatitis B Virus --- p.2 / Chapter 1.1.3 --- Hepatitis B Virus Infection around the World --- p.4 / Chapter 1.1.4 --- Hepatitis B Virus Infection in Hong Kong --- p.5 / Chapter 1.1.5 --- Hepatitis B Virus Infection in China --- p.7 / Chapter 1.1.5.1 --- Update of HBV Infection in China --- p.7 / Chapter 1.1.5.2 --- Problems in China --- p.7 / Chapter 1.2 --- Hepatitis B Virology --- p.8 / Chapter 1.2.1 --- Hepadnaviridae Family --- p.8 / Chapter 1.2.2 --- HBV Particles Types --- p.9 / Chapter 1.2.3 --- The HBV Genome --- p.10 / Chapter 1.2.4 --- The Life Cycle of HBV --- p.12 / Chapter 1.2.5 --- Hepatitis B Surface Antigen (HBsAg) --- p.17 / Chapter 1.3 --- HBV Transmission --- p.19 / Chapter 1.4 --- HBV Therapy --- p.19 / Chapter 1.5 --- Phyllanthus Species --- p.22 / Chapter 1.6 --- Alexander Cells --- p.26 / Chapter 1.7 --- Objectives --- p.29 / Chapter CHAPTER 2 --- COMPARISONS OF AQUEOUS AND ORGANIC EXTRACTS OF THREE PHYLLANTHUS SPECIES OF THEIR IN VITRO ANTI-HBV EFFECTS --- p.30 / Chapter 2.1 --- Introduction --- p.30 / Chapter 2.2 --- Materials and Methods --- p.30 / Chapter 2.2.1 --- Materials --- p.30 / Chapter 2.2.1.1 --- Phyllanthus species --- p.30 / Chapter 2.2.1.2 --- "Chemicals, Antibodies and Instrument" --- p.31 / Chapter 2.2.2 --- Extraction Methods --- p.32 / Chapter 2.2.2.1 --- Aqueous Extraction --- p.33 / Chapter 2.2.2.2 --- Organic Extraction --- p.33 / Chapter 2.2.3 --- Cell line --- p.33 / Chapter 2.2.4 --- Toxicity of Extracts --- p.34 / Chapter 2.2.5 --- IMx Assay --- p.34 / Chapter 2.2.6 --- Semi-quantitative RT-PCR --- p.35 / Chapter 2.2.6.1 --- RNA Extraction --- p.35 / Chapter 2.2.6.2 --- RT-PCR --- p.36 / Chapter 2.2.7 --- Western Blotting --- p.37 / Chapter 2.2.7.1 --- Preparation of Protein Samples --- p.37 / Chapter 2.2.7.2 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.37 / Chapter 2.2.7.3 --- Protein Transfer --- p.38 / Chapter 2.2.7.4 --- Immumnoblotting --- p.39 / Chapter 2.2.7.5 --- Protein Assay --- p.39 / Chapter 2.3 --- Results --- p.40 / Chapter 2.3.1 --- Toxicity of the Extracts --- p.40 / Chapter 2.3.2 --- Effects on HBsAg Secretion and Viral Gene Expression --- p.45 / Chapter 2.3.2 --- Analysis of Intracellular Viral Proteins --- p.58 / Chapter 2.4 --- Discussion --- p.63 / Chapter CHAPTER 3 --- ISOLATION AND CHARACTERIZATION OF ACTIVE COMPONIENT FROM AN ORGANIC EXTRACT OF PHYLLANTHUS URINARIA (GUANGDONG) --- p.68 / Chapter 3.1 --- Introduction --- p.68 / Chapter 3.2 --- Materials and Methods --- p.69 / Chapter 3.2.1 --- Materials --- p.69 / Chapter 3.2.2 --- Methods --- p.70 / Chapter 3.2.2.1 --- Ethanol Extraction --- p.70 / Chapter 3.2.2.2 --- Partitions --- p.70 / Chapter 3.2.2.3 --- Column Purification --- p.71 / Chapter 3.2.2.4 --- Analytical Thin Layer Chromatographic (TLC) --- p.71 / Chapter 3.2.2.5 --- Crystallization --- p.71 / Chapter 3.3 --- Results --- p.72 / Chapter 3.3.1 --- Analysis of Four Fractions after Partitions --- p.72 / Chapter 3.3.2 --- Screening of the Active Fraction after Column Chromatography of Fraction B --- p.76 / Chapter 3.3.3 --- Screening of the Active Fraction after Column Chromatography of Fraction6 --- p.79 / Chapter 3.3.4 --- Crystallization and Identification of the Isolated component --- p.82 / Chapter 3.3.5 --- Study Anti-HBV effects of pheophorbide a --- p.91 / Chapter 3.4 --- Discussion --- p.97 / Chapter CHAPTER 4 --- STUDY OF PRE S I PROMOTER ACTIVITY OF HBV --- p.103 / Chapter 4.1 --- Introduction --- p.103 / Chapter 4.2 --- Materials and Methods --- p.108 / Chapter 4.2.1 --- Materials --- p.108 / Chapter 4.2.2 --- Methods --- p.109 / Chapter 4.2.2.1 --- Cell line --- p.109 / Chapter 4.2.2.2 --- Clonning of Pre SI Promoter from HBV Genome --- p.109 / Chapter 4.2.2.3 --- Gene Clean --- p.110 / Chapter 4.2.2.4 --- Restriction Enzyme Digestion --- p.111 / Chapter 4.2.2.5 --- Synthesis of T-Overhang EcoR V Cut pBluescript® II KS (-) --- p.111 / Chapter 4.2.2.6 --- Ligation --- p.112 / Chapter 4.2.2.7 --- DH5α Competent Cells Preparation --- p.112 / Chapter 4.2.2.8 --- Transformation --- p.113 / Chapter 4.2.2.9 --- Plasmid Purification --- p.113 / Chapter 4.2.2.10 --- Transfection --- p.114 / Chapter 4.2.2.11 --- Luciferase Assay --- p.115 / Chapter 4.3 --- Results --- p.116 / Chapter 4.3.1 --- Cloning of the Pre S I Promoter --- p.116 / Chapter 4.3.2 --- Sequences of the Pre S I Promoter --- p.121 / Chapter 4.3.3 --- Pre S I Promoter Activities in Hep 3B Cell Line --- p.123 / Chapter 4.3.4 --- Effects of Herbal Extracts on Pre S I Promoter --- p.126 / Chapter 4.4 --- Discussion --- p.130 / Chapter CHAPTER 5 --- GENERAL DISCUSSION --- p.134 / REFERENCES --- p.141

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324699
Date January 2004
ContributorsLam, Kit., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xx, 153 leaves : ill. (some col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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