Wu-shun, Felix Wong. / Thesis (M.D.)--Chinese University of Hong Kong, 1991. / Includes bibliographies. / Title page --- p.i / Table of contents --- p.ii / Acknowledgements --- p.vi / Abstract --- p.viii / Declaration --- p.xiii / List of abbreviation --- p.xiv / Chapter Chapter one： --- Introduction --- p.1 / Chapter 1.1 --- Overview --- p.2 / Chapter 1.2 --- Aims of the study --- p.5 / Chapter Chapter two： --- Literature review --- p.8 / Chapter 2.1 --- Cell cycle and tumour growth --- p.9 / Chapter 2.1.1 --- Cell cycle --- p.9 / Chapter 2.1.2 --- Tumour growth --- p.14 / Chapter 2.2 --- Kinetic studies --- p.21 / Chapter 2.2.1 --- Radioisotopic studies --- p.21 / Chapter 2.2.2 --- Flow cytometry --- p.28 / Chapter 2.2.3 --- Monoclonal antibody --- p.32 / Chapter 2.3 --- Monoclonal antibody Ki-67 --- p.39 / Chapter 2.3.1 --- Development of Ki-67 --- p.39 / Chapter 2.3.2 --- The nature of the Ki-67 antigen --- p.42 / Chapter 2.3.3 --- Comparison with other kinetic methods --- p.45 / Chapter 2.3.4 --- Reported studies --- p.50 / Chapter 2.4 --- immunocytochemical staining --- p.63 / Chapter 2.4.1 --- Principle of immunostaining --- p.63 / Chapter 2.4.2 --- Fixation and processing methods --- p.69 / Chapter Chapter three： --- Materials and methods --- p.75 / Chapter 3.1 --- Cell culture --- p.76 / Chapter 3.1.1 --- Culture medium --- p.76 / Chapter 3.1.2 --- Origin and maintenance of cell line --- p.76 / Chapter 3.1.3 --- Coversip monolayer culture --- p.80 / Chapter 3.1.4 --- Multicellular spheroid culture --- p.80 / Chapter 3.1.5 --- Growth curve study --- p.81 / Chapter 3.1.6 --- Cytocentrifuge slide preparation --- p.81 / Chapter 3.2 --- immunoperoxidase staining --- p.83 / Chapter 3.2.1 --- Materials of immunoperoxidase staining --- p.83 / Chapter 3.2.2 --- Immunoperoxidase staining method --- p.86 / Chapter 3.3 --- Cell counting method --- p.92 / Chapter 3.3.1 --- Interactive cell counting system --- p.92 / Chapter 3.3.2 --- Cell counting methods --- p.95 / Chapter Chapter four： --- Proliferative activities of tumour cells IN VITRO --- p.104 / Chapter 4.1 --- Identification of cell proliferation of B16 melanoma cellsin VITRO --- p.105 / Chapter 4.1.1. --- Materials and methods --- p.106 / Chapter 4.1.2. --- Results --- p.107 / Chapter 4.1.3 --- Discussion --- p.110 / Chapter 4.2 --- Staining patterns of proliferating OCC1 cells in vitro --- p.117 / Chapter 4.2.1 --- Materials and methods --- p.117 / Chapter 4.2.2 --- Results --- p.118 / Chapter 4.2.3 --- Discussion --- p.121 / Chapter 4.3 --- "Comparative in vitro studies of cell proliferation using AgNOR counts, anti-BrdU, AD203 and Ki-67" --- p.130 / Chapter 4.3.1. --- Materials and methods --- p.130 / Chapter 4.3.2. --- Results --- p.131 / Chapter 4.3.3 --- Discussion --- p.134 / Chapter 4.4 --- Proliferative activities of tumor cells in vitro --- p.139 / Chapter 4.4.1. --- Materials and methods --- p.140 / Chapter 4.4.2. --- Results --- p.141 / Chapter 4.4.3 --- Discussion --- p.146 / Chapter Chapter five： --- Growth fraction in human genital tissues --- p.156 / Chapter 5.1 --- Cell proliferation in normal and neoplastic cervical tissues --- p.157 / Chapter 5.1.1. --- Materials and methods --- p.158 / Chapter 5.1.2. --- Results --- p.161 / Chapter 5.1.3 --- Discussion --- p.154 / Chapter 5.2 --- Tumour growth fraction in cervical carcinoma --- p.172 / Chapter 5.2.1. --- Materials and methods --- p.172 / Chapter 5.2.2. --- Results --- p.173 / Chapter 5.2.3 --- Discussion --- p.177 / Chapter 5.3 --- Tumour growth fraction in ovarian carcinoma --- p.185 / Chapter 5.3.1. --- Materials and methods --- p.185 / Chapter 5.3.2. --- Results --- p.186 / Chapter 5.3.3 --- Discussion --- p.190 / Chapter Chapter six： --- Conclusion --- p.198 / Chapter 6.1 --- Overview and future work --- p.199 / Chapter 6.2 --- Conclusion --- p.211 / references --- p.213 / Appendix： --- p.246 / Chapter (A) --- Additional Experiments / Chapter Experiment 1 --- Highest selection counting method --- p.246 / Chapter Experiment 2 --- Double staining of B16 melanoma cells --- p.248 / Chapter Experiment 3 --- Trypan blue exclusion test for viability --- p.250 / Chapter (B) --- Selected publications by the author / Chapter Publication 1 --- Characteristics of a cell line established from a Chinese patient with a squamous carcinoma of the uterine cervix --- p.252 / Chapter Publication 2 --- Establishment and characterization of a new human cell line derived from ovarian clear cell carcinoma --- p.258 / Chapter Publication 3 --- "Identification of ""non-proliferating"" B16 melanoma cells using monoclonal antibody (AD203) against the Ml subunit of ribonucleotide reductase" --- p.267 / Chapter Publication 4 --- The correlation of agyrophilic nucleolar organiser regions (AgNORs) count to bromodeoxyuridine incorporation and Ki-67 scores in an ovarian carcinoma cell line --- p.275 / Chapter Publication 5 --- Immunohistochemical determination of tumour growth fraction in human ovarian carcinoma --- p.278 / Chapter Publication 6 --- Tumor growth fraction in cervical carcinoma --- p.283

by Po Chu, Leung. / Thesis submitted in: December 1999. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 141-169). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abbreviation --- p.iii / Abstract --- p.vii / Chinese Abstract --- p.x / Table of Contents --- p.xii / Chapter Chapter 1: --- General Introduction / Chapter 1.1 --- Hematopoiesis --- p.1 / Chapter 1.1.1 --- The Development of Hematopoietic Progenitor Cells --- p.1 / Chapter 1.1.2 --- Hematopoietic Growth Factors --- p.3 / Chapter 1.1.3 --- Transcriptionl Factors Involved in Lineage Commitment of Hematopoietic Progenitor Cells --- p.5 / Chapter 1.2 --- Leukemia --- p.7 / Chapter 1.2.1 --- Occurrence and Classification of Leukemia --- p.7 / Chapter 1.2.2 --- The Pathological Features and Etiology of Leukemia --- p.10 / Chapter 1.2.3 --- The Molecular Basis of Leukemia --- p.13 / Chapter 1.2.4 --- Current Therapeutic Strategies --- p.14 / Chapter 1.2.4.1 --- Conventional Therapies for Leukemia --- p.14 / Chapter 1.2.4.2 --- Induction of Cell Differentiation and Apoptosis for Treatment of Leukemia --- p.16 / Chapter 1.2.5 --- The Use of Murine Myelomonocytic Leukemia WEHI-3B JCS Cells As a Model for the Study of Leukemia Cell Proliferation， Differentiation and Survival --- p.22 / Chapter 1.3 --- Taxol: A Novel Anti-cancer Agent --- p.23 / Chapter 1.3.1 --- Discovery and Action Mechanism --- p.23 / Chapter 1.3.2 --- Metabolism and Toxicity of Taxol --- p.27 / Chapter 1.3.3 --- The Biological Activities of Taxol --- p.28 / Chapter 1.3.4 --- The Anti-tumor Effects of Taxol --- p.30 / Chapter 1.3.5 --- The Effects of Taxol on Leukemia --- p.31 / Chapter 1.4 --- Aims and Scopes of This Investigation --- p.32 / Chapter Chapter 2: --- Materials and Methods / Chapter 2.1 --- Materials --- p.35 / Chapter 2.1.1 --- Mice --- p.35 / Chapter 2.1.3 --- "Culture Media,Buffer and Other Solutions" --- p.37 / Chapter 2.1.4 --- Radioisotope and Scintillation Fluid --- p.39 / Chapter 2.1.5 --- Taxol --- p.40 / Chapter 2.1.6 --- Recombinant Cytokines --- p.40 / Chapter 2.1.7 --- Vitamin Analogs --- p.42 / Chapter 2.1.8 --- Various Signal Transduction Pathway Activators and Inhibitors --- p.42 / Chapter 2.1.9 --- Monoclonal Antibodies and Buffers for Flow Cytometry --- p.43 / Chapter 2.1.10 --- Reagents and Chemicals for Gene Expression Study --- p.45 / Chapter 2.1.11 --- Chemical Solutions and Buffers for Western Blot --- p.50 / Chapter 2.1.12 --- Reagents for Colony Assay --- p.54 / Chapter 2.2 --- Methods --- p.55 / Chapter 2.2.1 --- Culture of Leukemia Cell Lines --- p.55 / Chapter 2.2.2 --- Treatment of Leukemia Cells with Various Drugs and Cytokines --- p.55 / Chapter 2.2.3 --- Cell Morphological Study --- p.55 / Chapter 2.2.4 --- Determination of Leukemia Cell Survival and Proliferation --- p.56 / Chapter 2.2.5 --- Colony Assay --- p.56 / Chapter 2.2.6 --- Flow Cytometry Analysis --- p.57 / Chapter 2.2.6.1 --- Surface Antigen Immunophenotyping --- p.57 / Chapter 2.2.6.2 --- Assay of Endocytic Activity --- p.58 / Chapter 2.2.6.3 --- Cell Cycle /DNA Content Evaluation --- p.58 / Chapter 2.2.7 --- Gene Expression Study --- p.59 / Chapter 2.2.7.1 --- Preparation of Total Cellular RNA --- p.59 / Chapter 2.2.7.2 --- Reverse Transcription --- p.60 / Chapter 2.2.7.3 --- Polymerase Chain Reaction (PCR) --- p.60 / Chapter 2.2.7.4 --- Agarose Gel Electrophoresis --- p.61 / Chapter 2.2.8 --- DNA Fragmentation Analysis --- p.61 / Chapter 2.2.9 --- Protein Expression Study --- p.62 / Chapter 2.2.9.1 --- Protein Extraction --- p.62 / Chapter 2.2.9.2 --- Quantification of the Protein --- p.62 / Chapter 2.2.9.3 --- Western Blot Analysis --- p.63 / Chapter 2.2.10 --- Statistical Analysis --- p.64 / Chapter Chapter 3: --- Results / Chapter 3.1 --- Effects of Taxol on the Proliferation and Apoptosis of the Murine Myeloid Leukemia Cells --- p.65 / Chapter 3.1.1 --- Growth-Inhibitory Effects of Taxol on Murine Myeloid Leukemia WEHI-3B JCS cells --- p.65 / Chapter 3.1.2 --- Cytotoxic Effects of Taxol on Murine Bone Marrow Cells and Myeloid Leukemia WEHI-3B JCS Cells --- p.69 / Chapter 3.1.3 --- Anti-proliferative Effect of Taxol on Different Leukemia Cell Lines --- p.72 / Chapter 3.1.4 --- Effects of Taxol on the Cell Cycle Kinetics of WEHI-3B JCS Cells --- p.81 / Chapter 3.1.5 --- Induction of DNA Fragmentation of WEHI-3B JCS cells by Taxol --- p.83 / Chapter 3.1.6 --- Effect of Taxol on the Clonogenicity of WEHI-3B JCS Cells In Vitro and Tumorigenicity In Vivo --- p.86 / Chapter 3.2 --- Effects of Taxol on the Induction of Monocytic Cell Differentiation in Murine Myeloid Leukemia Cells --- p.88 / Chapter 3.2.1 --- Morphological Changes in Taxol-Treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.88 / Chapter 3.2.2 --- Surface Antigen Immunophenotyping of Taxol-treated WE HI-3B cells --- p.91 / Chapter 3.2.3 --- Endocytic Activity of Taxol-treated WEHI-3B JCS cells --- p.95 / Chapter 3.3 --- Modulatory Effect of Taxol and Cytokines on the Proliferation of WEHI- 3B JCS Cells --- p.96 / Chapter 3.4 --- Modulatory Effect of Taxol and Physiological Differentiation Inducers on the Proliferation of WEHI-3B JCS cells --- p.103 / Chapter 3.5 --- The Possible Involvement of Protein Kinase C in the Anti-proliferative Activity of Taxol on WEHI-3B JCS Cells --- p.106 / Chapter 3.6 --- Modulation of Apoptotic Gene Expression in Taxol-treated WEHI-3B JCS cells --- p.113 / Chapter 3.7 --- Modulatory Effects of Taxol on the Protein Expression of WEHI-3B JCS Cells --- p.119 / Chapter Chapter 4: --- Discussion and Conclusions / Chapter 4.1 --- "Effects of Taxol on the Proliferation,Differentiation and Apoptosis of the Murine Myeloid Leukemia Cells" --- p.126 / Chapter 4.2 --- "Modulatory Effects of Taxol, Cytokines and Physiological Differentiation Inducers on the Proliferation of the Myelomonocytic Leukemia WEHI-3B JCS Cells" --- p.132 / Chapter 4.3 --- The Possible Involvement of Protein Kinase C in Anti-proliferative Activity of Taxol on WEHI-3B JCS Cells --- p.136 / Chapter 4.4 --- The Modulation of Apoptosis Gene Expression in Taxol-treated WEHI-3B JCS Cells --- p.137 / Chapter 4.5 --- The Modulation of Protein Expression in Taxol-treated WEHI-3B JCS Cells --- p.138 / Chapter 4.6 --- Conclusions and Future Perspectives --- p.139 / References --- p.141

Yeung Chi Hang, Thomas. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 126-146). / Abstracts in English and Chinese. / Table of content / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / LIST OF ABBREVIATION --- p.vi / TABLE OF CONTENTS --- p.vii / Chapter Chapter1 --- General Introduction / Chapter 1.1 --- INTRODUCTION --- p.1 / Chapter 1.2 --- FORMATION OF CLA --- p.1 / Chapter 1.3 --- OCCURRENCE OF CLA IN FOODS --- p.5 / Chapter 1.4 --- PHYSIOLOGICAL EFFECTS OF CLA --- p.8 / Chapter 1.4.1 --- Anticarcinogenic Effects of CLA --- p.8 / Chapter 1.4.2 --- Antiatherogenic Effect of CLA --- p.11 / Chapter 1.4.3 --- Antioxidant Effect of CLA --- p.12 / Chapter 1.4.4 --- CLA and Immune Response --- p.13 / Chapter 1.4.5 --- CLA and Body Composition --- p.14 / Chapter Chapter2 --- Protective Effect of CLA on Copper-Induced Human LDL Oxidation / Chapter 2.1 --- INTRODUCTION --- p.17 / Chapter 2.1.1 --- Oxidative Modification of LDL and Atherosclerosis --- p.20 / Chapter 2.1.1.1 --- Understanding of LDL --- p.20 / Chapter 2.1.1.2 --- Oxidative Modification of LDL --- p.20 / Chapter 2.1.1.3 --- Role of Oxidative Modified LDL in Atherogenesis --- p.23 / Chapter 2.1.2 --- Antioxidants and Atherosclerosis --- p.25 / Chapter 2.1.3 --- Measuring TBARS Formation as an in vitro Index to Monitor LDL Oxidation --- p.26 / Chapter 2.1.4 --- CLA and Atherogenesis --- p.27 / Chapter 2.2 --- OBJECTIVE OF THE PRESENT STUDY --- p.28 / Chapter 2.3 --- MATERIALS AND METHODS --- p.29 / Chapter 2.3.1 --- Human LDL Isolation --- p.29 / Chapter 2.3.2 --- LDL Oxidation --- p.30 / Chapter 2.3.3 --- Thiobarbituric Acid Reactive Substances (TBARS) Assay --- p.30 / Chapter 2.4 --- STATISTICS --- p.31 / Chapter 2.5 --- RESULTS --- p.32 / Chapter 2.5.1 --- Inhibitory Effect of BSA on Human LDL Oxidation --- p.32 / Chapter 2.5.2 --- Pro-oxidant Effect of LA on Human LDL Oxidation --- p.32 / Chapter 2.5.3 --- Inhibitory Effect of CLA on Human LDL Oxidation --- p.32 / Chapter 2.6 --- DISCUSSION --- p.37 / Chapter 2.6.1 --- Effect ofBSA on Copper-Induced LDL Oxidation --- p.37 / Chapter 2.6.2 --- Effect of LA on Copper-Induced LDL Oxidation --- p.38 / Chapter 2.6.3 --- Protective Effect of CLA on Copper-Induced Human LDL Oxidation --- p.39 / Chapter Chapter3 --- Hypolipidemic Activity of CLA / Chapter 3.1 --- INTRODUCTION --- p.42 / Chapter 3.1.1 --- Total Cholesterol and LDL Cholesterol --- p.42 / Chapter 3.1.2 --- Triglyceride (TG) --- p.44 / Chapter 3.1.3 --- Hypolipidemic Effect of CLA --- p.45 / Chapter 3.1.4 --- Golden Syrian Hamster as an Animal Model of Cholesterol Metabolism --- p.46 / Chapter 3.2 --- OBJECTIVES OF THE PRESENT STUDY --- p.48 / Chapter 3.3 --- MATERIALS AND METHODS --- p.48 / Chapter 3.3.1 --- LA and CLA --- p.49 / Chapter 3.3.2 --- Animals --- p.49 / Chapter 3.3.3 --- Experiment1 --- p.49 / Chapter 3.3.4 --- Experiment2 --- p.51 / Chapter 3.3.5 --- "Determination of Serum TC, HDL-Cholesterol (HDL-C) and TG" --- p.54 / Chapter 3.3.6 --- Lipid analysis of Liver and Adipose Tissue --- p.54 / Chapter 3.3.6.1 --- Lipid Extraction and Separation of Different Lipid Species --- p.54 / Chapter 3.3.6.2 --- Acid-Catalyzed Methylation of Fatty Acids --- p.55 / Chapter 3.3.6.3 --- GLC Analysis of FAME --- p.55 / Chapter 3.3.7 --- Quantification of Tissue Cholesterol --- p.56 / Chapter 3.3.7.1 --- Cholesterol Extraction and Silylation --- p.56 / Chapter 3.3.7.2 --- GLC Analysis of TMS-Ether Derivative of Cholesterol --- p.56 / Chapter 3.4 --- STATISTICS --- p.57 / Chapter 3.5 --- RESULTS --- p.59 / Chapter 3.5.1 --- Body Weight and Food Intake --- p.59 / Chapter 3.5.2 --- "Effect of Dietary CLA Supplementation on Serum TG, TC and HDL-C" --- p.59 / Chapter 3.5.3 --- "Effect of Dietary CLA Supplementation on Hepatic TG, Phospholipid and Cholesterol" --- p.64 / Chapter 3.5.4 --- Effect of Dietary CLA Supplementation on Adipose Tissue TG and Cholesterol --- p.73 / Chapter 3.5.5 --- Effect of CLA Supplementation on Cholesterol Levels of Different Tissues --- p.73 / Chapter 3.6 --- DISCUSSION --- p.79 / Chapter 3.6.1 --- "Effect of CLA Supplementation on Serum TG, TC and HDL-C" --- p.79 / Chapter 3.6.2 --- "Effect of CLA Supplementation on Hepatic TG, PL and Cholesterol" --- p.81 / Chapter 3.6.3 --- Effect of CLA on Adipose Tissue TG and Cholesterol --- p.83 / Chapter 3.6.4 --- Implication of CLA Intake in Humans --- p.84 / Chapter Chapter4 --- Influences of Dietary CLA on Cholesterol Homeostasis / Chapter 4.1 --- INTRODUCTION --- p.86 / Chapter 4.2 --- NEUTRAL EFFECT OF DIETARY CLA SUPPLEMENTATION ON HMG-COA REDUCTASE ACTIVITY --- p.88 / Chapter 4.2.1 --- HMG-CoA Reductase as the Rate-Limiting Enzyme in Cholesterol Synthesis --- p.88 / Chapter 4.2.2 --- Objective of The Present Study --- p.91 / Chapter 4.2.3 --- Materials and Methods --- p.92 / Chapter 4.2.3.1 --- Preparation of Hepatic Microsome --- p.92 / Chapter 4.2.3.2 --- HMG-GoA Reductase Activity Assay --- p.92 / Chapter 4.2.4 --- Statistics --- p.93 / Chapter 4.2.5 --- Results --- p.94 / Chapter 4.2.6 --- Discussion --- p.96 / Chapter 4.3 --- DOWN-REGULATION OF THE INTESTINAL ACAT ACTIVITY BY CLA FEEDING --- p.97 / Chapter 4.3.1 --- Role of ACAT in Cholesterol Absorption --- p.97 / Chapter 4.3.2 --- Objective of The Present Study --- p.99 / Chapter 4.3.3 --- Materials and Methods --- p.100 / Chapter 4.3.3.1 --- Preparation of Intestinal Microsome --- p.100 / Chapter 4.3.3.2 --- ACAT Activity Assay --- p.100 / Chapter 4.3.4 --- Statistics --- p.101 / Chapter 4.3.5 --- Results --- p.102 / Chapter 4.3.6 --- Discussion --- p.104 / Chapter 4.4 --- ALTERATION OF FECAL EXCRETION BY DIETARY CLA --- p.105 / Chapter 4.4.1 --- Objective of The Present Study --- p.108 / Chapter 4.4.2 --- Materials and Methods --- p.109 / Chapter 4.4.2.1 --- Separation of Neutral and Acidic Sterols --- p.109 / Chapter 4.4.2.2 --- Neutral Sterol Analysis --- p.109 / Chapter 4.4.2.3 --- Acidic Sterol Analysis --- p.110 / Chapter 4.4.2.4 --- GLC Analysis of Neutral and Acidic Sterols --- p.110 / Chapter 4.4.3 --- Statistics --- p.113 / Chapter 4.4.4 --- Results --- p.114 / Chapter 4.4.4.1 --- Effect of CLA Supplementation on Fecal Output of Neutral Sterols --- p.114 / Chapter 4.4.4.2 --- Effect of CLA Supplementation on Fecal Output of Acidic Sterols --- p.114 / Chapter 4.4.5 --- Discussion --- p.118 / Chapter Chapter5 --- Conclusions --- p.123 / References --- p.126

by Tsui Hong Teng. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 174-181). / Abstracts in English and Chinese. / A CKNO WLED GMENTS --- p.7 / ABSTRACT --- p.8-10 / Chapter Chapter 1: --- Introduction: --- p.11-44 / Chapter 1.1) --- Glucose transporters / Chapter 1.2) --- Glucose transporters and cancers / Chapter 1.3) --- Antisense strategies / Chapter 1.4) --- Cellular uptake of oligonucleotides / Chapter 1.5) --- Hyperthermia and combined treatments / Chapter Chapter 2: --- Materials and methods --- p.45-60 / Chapter 2.1) --- Materials: / Chapter 2.1a) --- Cell lines and culture media / Chapter 2.1b) --- Oligonucleotides synthesis / Chapter 2.1c) --- Chemicals / Chapter 2.2) --- Methods: / Chapter 2.2a) --- Oligonucleotide design / Chapter 2.2b) --- Oligonucleotide treatment / Chapter 2.2c) --- Flow cytometry / Chapter 2.2d) --- Confocal microscopy / Chapter 2.2e) --- MTT assay for cytotoxicity or cell proliferation / Chapter Chapter 3: --- Cellular uptake of oligonucleotide spontaneously and Lipofectin-aided: --- p.61-85 / Chapter 3.1) --- Introduction / Chapter 3.2) --- Flow cytometric studies / Chapter 3.3) --- Confocal microscopic studies / Chapter 3.4) --- Cytotoxic effect of Lipofectin alone on MCF-7 cells / Chapter 3.5) --- Discussion / Chapter Chapter 4: --- Hyperthermia can enhance oligonucleotide uptake: --- p.86-118 / Chapter 4.1) --- Introduction / Chapter 4.2) --- Flow cytometric studies / Chapter 4.3) --- Confocal microscopic studies / Chapter 4.4) --- Cytotoxic effect of hyperthermia on MCF-7 cells / Chapter 4.5) --- FITC-ODN uptake in survival cells by propidium iodide (PI) exclusion method for hyperthermia / Chapter 4.6) --- Discussion / Chapter Chapter 5: --- The antiproliferative effects of antisense molecules against Glut-1 and 5 on MCF- 7 cells transfected by Lipofectin: --- p.119-146 / Chapter 5.1) --- Introduction / Chapter 5.2) --- The growth curve of MCF-7 cells / Chapter 5.3) --- The calibration of MTT assay / Chapter 5.4) --- The effect of antisense Glut-1 concentration without Lipofectin on MCF-7 cells / Chapter 5.5) --- The effect of antisense Glut-1 concentration with Lipofectin on MCF-7 cells / Chapter 5.6) --- The effect of antisense Glut-5 concentration without Lipofectin on MCF-7 cells / Chapter 5.7) --- The effect of antisense Glut-5 concentration with Lipofectin on MCF-7cells / Chapter 5.8) --- The effect of transfection time of antisense Glut-1 on MCF-7 cells / Chapter 5.9) --- The effect of transfection time of antisense Glut-5 on MCF-7 cells / Chapter 5.10) --- The effect of transfection time of antisense Glut-5 for higher concentration on MCF-7 cells / Chapter 5.11) --- The effect of antisense Glut-1 to Lipofectin (w/w) ratio on MCF-7 cells / Chapter 5.12) --- The effect of antisense Glut-1 to Lipofection (w/w) ratio for higher transfection time on MCF-7 cells / Chapter 5.13) --- The effect of antisense Glut-5 to Lipofectin (w/w) ratio on MCF-7 cells / Chapter 5.14) --- Discussion / Chapter Chapter 6: --- Combined treatments: --- p.147-162 / Chapter 6.1) --- Introduction / Chapter 6.2) --- The effect of combined treatment of antisense Glut-1 combined with antisense Glut-5 on MCF-7 cells / Chapter 6.3) --- The chronic effect of hyperthermia for 5 hours on MCF-7 cells / Chapter 6.4) --- The effect of combined treatment between antisense Glut-1 and hyperthermia on MCF-7 cells / Chapter 6.5) --- The net effect of antisense Glut-1 in combined treatment between hyperthermia and antisense Glut-1 on MCF-7 cells / Chapter 6.6) --- The effect of combined treatment between antisense Glut-5 and hyperthermia on MCF-7 cells / Chapter 6.7) --- The net effect of antisense Glut-5 in combined treatment between hyperthermia and antisense Glut-5 on MCF-7 cells / Chapter 6.8) --- Discussion / Chapter Chapter 7: --- Discussion: --- p.163-173 / Chapter Chapter 8: --- References: --- p.174-181

Siu Pak-yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 158-174). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Contents --- p.vi / List of Figures and Tables --- p.xiii / List of Abbreviations --- p.xviii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Characteristics of Arsenic Compound --- p.1 / Chapter 1.1.1 --- Arsenic Compounds are Used as Poison --- p.1 / Chapter 1.1.2 --- Arsenic Compounds are Used as Medicine --- p.2 / Chapter 1.2 --- Arsenic Trioxide is a Traditional Chinese Medicine --- p.3 / Chapter 1.3 --- Properties of Arsenic Trioxide --- p.5 / Chapter 1.4 --- Use of Arsenic Trioxide in Cancer Treatment --- p.7 / Chapter 1.4.1 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Acute Promyelocytic Leukemia --- p.7 / Chapter 1.4.1.1 --- Characteristics of Acute Promyelocytic Leukemia --- p.7 / Chapter 1.4.1.2 --- Treatment of Acute Promyelocytic Leukemia with All-Trans Retinoic Acid --- p.10 / Chapter 1.4.1.3 --- Treatment of Acute Promyelocytic Leukemia with Arsenic Trioxide --- p.11 / Chapter 1.4.1.4 --- Action Mechanism of Arsenic Trioxide --- p.13 / Chapter 1.4.2 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Non-APL Leukemia --- p.15 / Chapter 1.4.3 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Solid Tumors --- p.16 / Chapter 1.5 --- Human Hepatocellular Carcinoma --- p.16 / Chapter 1.5.1 --- The Incidence of Liver Cancer --- p.16 / Chapter 1.5.2 --- Classification of Liver Cancer --- p.17 / Chapter 1.6 --- Aim of the Project --- p.17 / Chapter 1.6.1 --- In Vitro Study of the Effect of Arsenic Trioxide on HepG2 Cells --- p.19 / Chapter 1.6.2 --- In Vivo Study of the Effect of Arsenic Trioxide by Tumor-Bearing Nude Mice Model --- p.20 / Chapter 1.6.3 --- "In Vitro Study of the Effect of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" --- p.22 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.24 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.24 / Chapter 2.1.1.1 --- Cell Lines --- p.24 / Chapter 2.1.1.2 --- Culture Medium --- p.25 / Chapter 2.1.2 --- Chemicals --- p.26 / Chapter 2.1.3 --- Reagents and Buffers --- p.27 / Chapter 2.1.3.1 --- Phosphate Buffered Saline (PBS) --- p.27 / Chapter 2.1.3.2 --- "3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) Solution" --- p.27 / Chapter 2.1.3.3 --- Reagents for DNA Fragmentation Assay --- p.21 / Chapter 2.1.3.3.1 --- DNA Lysis Buffer --- p.27 / Chapter 2.1.3.3.2 --- Tris-EDTA (TE) Buffer --- p.27 / Chapter 2.1.3.3.3 --- Tris-Acetate (TAE) Buffer --- p.28 / Chapter 2.1.3.3.4 --- Proteinase K and Ribonuclease A (RNase A) --- p.28 / Chapter 2.1.3.3.5 --- 6X DNA Loading Dye --- p.28 / Chapter 2.1.3.3.6 --- One Hundred Base-Pair DNA Ladder --- p.28 / Chapter 2.1.3.4 --- Reagents for Western Blot Analysis --- p.29 / Chapter 2.1.3.4.1 --- SDS Lysis Buffer --- p.29 / Chapter 2.1.3.4.2 --- 4X Lower Gel Buffer --- p.29 / Chapter 2.1.3.4.3 --- 4X Upper Gel Buffer --- p.29 / Chapter 2.1.3.4.4 --- 10X SDS Running Buffer --- p.29 / Chapter 2.1.3.4.5 --- 2X SDS Sample Loading Dye --- p.30 / Chapter 2.1.3.4.6 --- Electroblotting Buffer --- p.30 / Chapter 2.1.3.4.7 --- Tris-Buffered Saline with 01% Tween-20 (TBS-T) --- p.30 / Chapter 2.1.3.4.8 --- Lysis Buffer for Detection of the Release of Cytochrome C --- p.31 / Chapter 2.1.3.5 --- Propidium Iodide (PI) --- p.31 / Chapter 2.1.3.6 --- "5，5 ´ة,6,6´ة-tetrachloro-1,1',3,3 '-tetraethylbenzimidazolyl carbocyanine Iodide (JC-1)" --- p.31 / Chapter 2.1.3.7 --- Reagents for In Vivo Study --- p.32 / Chapter 2.1.3.7.1 --- Saline --- p.32 / Chapter 2.1.3.7.2 --- Homogenizing Buffer --- p.32 / Chapter 2.1.3.7.3 --- 10% Buffered Formalin --- p.32 / Chapter 2.1.3.7.4 --- Acid Alcohol --- p.32 / Chapter 2.1.3.7.5 --- Scott's Tap Water --- p.32 / Chapter 2.1.3.7.6 --- 0.5% Aqueous Eosin --- p.33 / Chapter 2.2 --- Methods --- p.33 / Chapter 2.2.1 --- MTT Assay --- p.33 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.34 / Chapter 2.2.3 --- Analysis of Cell-Cycle Phase Distribution by Flow Cytometry with PI Staining --- p.34 / Chapter 2.2.4 --- DNA Fragmentation Assay --- p.35 / Chapter 2.2.5 --- Quantification of Apoptosis by Flow Cytometry with Annexin V-PI Staining --- p.36 / Chapter 2.2.6 --- Assessment of the Change in Mitochondrial Membrane Potential (ΔΦm) --- p.37 / Chapter 2.2.7 --- Western Analysis --- p.38 / Chapter 2.2.8 --- Glucose Uptake Assay --- p.40 / Chapter 2.2.9 --- ATP Production Assay --- p.41 / Chapter 2.2.10 --- In Vivo Study --- p.44 / Chapter 2.2.10.1 --- Animal Model --- p.44 / Chapter 2.2.10.2 --- Cell Line --- p.44 / Chapter 2.2.10.3 --- Treatment with Arsenic Trioxide --- p.44 / Chapter 2.2.10.4 --- Assessment of the Anti-Cancer Activity of Arsenic Trioxide --- p.45 / Chapter 2.2.10.5 --- Tissue Sample Preparation --- p.45 / Chapter 2.2.10.5.1 --- Preparation of Plasma --- p.45 / Chapter 2.2.10.5.2 --- Preparation of Liver Tissue Homogenate --- p.46 / Chapter 2.2.10.5.3 --- Preparation of Cytosolic Fraction --- p.46 / Chapter 2.2.10.6 --- Measurement of the Plasma Enzyme Activity --- p.46 / Chapter 2.2.10.6.1 --- "Plasma Creatine Kinase (CK) Activity, Plasma Lactate Dehydrogenase (LDH) Activity, Plasma Alanine Transaminase (ALT) Activity and Plasma Asparate Transaminase (AST) Activity" --- p.46 / Chapter 2.2.10.7 --- Preparation of Tissue for Light Microscopic Study --- p.48 / Chapter 2.2.10.8 --- Measurement of the Basal Reduced Glutathione (GSH) Level of Liver Tissue --- p.51 / Chapter 2.2.10.9 --- "Measurement of the Activity of Antioxidant Enzyme, Glutathione S-Transferase (GST) of Liver Tissue" --- p.53 / Chapter 2.3 --- Statistical Analysis --- p.54 / Chapter Chapter 3 --- "In Vitro Study of Arsenic Trioxide on Acute Promyelocytic Leukemia Cell Line, NB-4" / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Principle of Flow Cytometry with Annexin V-PI Staining --- p.56 / Chapter 3.3 --- The Effect of Arsenic Trioxide on Cell Proliferation of NB-4 Cells --- p.59 / Chapter 3.4 --- Study of the Action Mechanism of Arsenic Trioxide upon Treatment of NB-4 Cells --- p.61 / Chapter 3.5 --- Summary --- p.63 / Chapter Chapter 4 --- "In Vitro Study of Arsenic Trioxide on Human Hepatocellular Carcinoma Cell Line, HepG2" / Chapter 4.1 --- Introduction --- p.64 / Chapter 4.2 --- The Effect of Arsenic Trioxide on Cell Proliferation of HepG2 Cells by MTT Assay --- p.66 / Chapter 4.3 --- The Effect of Arsenic Trioxide on HepG2 Cells at Clinically Achievable Concentration --- p.68 / Chapter 4.3.1 --- The Cytotoxicity of Arsenic Trioxide on HepG2 Cells by Trypan Blue Exclusion Assay --- p.68 / Chapter 4.3.2 --- The Effect of Arsenic Trioxide on Cell-Cycle Phase Distribution --- p.71 / Chapter 4.3.3 --- The Underlying Mechanism of the Cytotoxic Effect of Arsenic Trioxide 一 Necrosis or Apoptosis? --- p.74 / Chapter 4.3.3.1 --- DNA Fragmentation Assay --- p.74 / Chapter 4.3.3.2 --- Flow Cytometry with Annexin V-PI Staining --- p.76 / Chapter 4.3.3.3 --- Brief Conclusion --- p.78 / Chapter 4.3.4 --- The Study of the Mechanism of Apoptotic Pathway --- p.78 / Chapter 4.3.4.1 --- Activation of Caspase-3 upon Arsenic Trioxide Treatment --- p.79 / Chapter 4.3.4.2 --- The Participation of Mitochondria in Arsenic Trioxide-Induced Apoptosis --- p.81 / Chapter 4.3.4.2.1 --- The Change in Mitochondrial Membrane Potential upon Arsenic Trioxide Treatment --- p.81 / Chapter 4.3.4.2.2 --- The Study of the Release of Cytochrome C from the Mitochondria to Cytosol upon Treatment with Arsenic Trioxide --- p.85 / Chapter 4.3.4.2.3 --- Brief Conclusion --- p.87 / Chapter 4.4 --- Arsenic Trioxide Mediated Its Effect via Other Action Mechanisms --- p.87 / Chapter 4.4.1 --- The Effect of Arsenic Trioxide on the Expression of Glucose Transporters 1 and2 --- p.88 / Chapter 4.4.2 --- The Effect of Arsenic Trioxide on Glucose Uptake --- p.91 / Chapter 4.4.3 --- The Effect of Arsenic Trioxide on ATP Production --- p.93 / Chapter 4.4.4 --- Brief Conclusion --- p.93 / Chapter 4.5 --- Summary --- p.95 / Chapter Chapter 5 --- In Vivo Study of Arsenic Trioxide on HepG2-Bearing Nude Mice / Chapter 5.1 --- Introduction --- p.96 / Chapter 5.2 --- Treatment with Arsenic Trioxide --- p.97 / Chapter 5.3 --- Assessment of the Anti-Tumor Effect of Arsenic Trioxide --- p.99 / Chapter 5.4 --- The Effect of Arsenic Trioxide toward Normal Tissues --- p.103 / Chapter 5.4.1 --- The Effect of Arsenic Trioxide on Liver --- p.104 / Chapter 5.4.1.1 --- Morphological Study --- p.104 / Chapter 5.4.1.2 --- Enzymatic Study --- p.107 / Chapter 5.4.1.3 --- Brief Conclusion --- p.107 / Chapter 5.4.2 --- The Effect of Arsenic Trioxide on Heart --- p.110 / Chapter 5.4.2.1 --- Morphological Study --- p.110 / Chapter 5.4.2.2 --- Enzymatic Study --- p.112 / Chapter 5.4.2.3 --- Brief Conclusion --- p.112 / Chapter 5.5 --- Involvement of the Glutathione Redox System --- p.115 / Chapter 5.5.1 --- Basal GSH Level --- p.115 / Chapter 5.5.2 --- The Activity of Glutathion S-Transferase --- p.117 / Chapter 5.5.3 --- Brief Conclusion --- p.117 / Chapter 5.6 --- Summary --- p.120 / Chapter Chapter 6 --- "In Vitro Study of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" / Chapter 6.1 --- Introduction --- p.121 / Chapter 6.2 --- The Effect of Doxorubicin on the Parental HepG2 Cells and R-HepG2 Cells by MTT Assay --- p.123 / Chapter 6.3 --- The Effect of Arsenic Trioxide on Cell Proliferation of R-HepG2 Cells by MTT Assay --- p.126 / Chapter 6.4 --- The Effect of Arsenic Trioxide on Cell-Cycle Phase Distribution of R-HepG2 Cells --- p.129 / Chapter 6.5 --- Trioxide on R-HepG2 Cells ´ؤ Necrosis or Apoptosis? --- p.131 / Chapter 6.5.1 --- DNA Fragmentation Assay --- p.131 / Chapter 6.5.2 --- Flow Cytometry with Annexin V-PI Staining --- p.133 / Chapter 6.5.3 --- Brief Conclusion --- p.133 / Chapter 6.6 --- Examination of the Probable Involvement of Arsenic Trioxide as a Substrate of P-Glycoprotein --- p.135 / Chapter 6.7 --- Summary --- p.137 / Chapter Chapter 7 --- Discussion / Chapter 7.1 --- The Significance of the Study of Arsenic Trioxide in the Treatment of Arsenic Trioxide --- p.138 / Chapter 7.2 --- Comparison of Preparation of Drug in Present Study with Others --- p.140 / Chapter 7.3 --- Effect of Arsenic Trioxide on Human Hepatocellular Carcinoma --- p.142 / Chapter 7.4 --- Mechanism Study of Arsenic Trioxide --- p.142 / Chapter 7.5 --- Dosage of Arsenic Trioxide Used in In Vivo Study --- p.152 / Chapter 7.6 --- Cytotoxicity of Arsenic Trioxide toward Normal Tissues --- p.153 / Chapter 7.7 --- "Effect of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" --- p.154 / Chapter 7.8 --- Conclusions and Future Prospect --- p.156 / Chapter Chapter 8 --- References / Chapter 8.1 --- English References --- p.158 / Chapter 8.2 --- Chinese References --- p.174 / Chapter 8.3 --- Online References --- p.174

Chan Ping Kei. / Thesis submitted in: December 2001. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 135-143). / Abstracts in English and Chinese. / ACKNOWLEGEMENTS --- p.i / ABSTRACT --- p.ii / TABLE OF CONTECTS --- p.v / LIST OF FIGURE --- p.viii / LIST OF ABBREVIATIONS --- p.xii / CHAPTER / Chapter 1 --- INTRODUCTION / Chapter 1.1 --- Epidemilogy --- p.1 / Chapter 1.2 --- Malaria disease --- p.1 / Chapter 1.3 --- Life cycle of Malaria --- p.1 / Chapter 1.4 --- Current measure to control Malaria --- p.6 / Chapter 1.5 --- Anti-malaria vaccine candidate --- p.7 / Chapter 1.6 --- Anti-erythrocytic malaria vaccine MSP-1 --- p.10 / Chapter 1.7 --- Baculovirus Expression System --- p.20 / Chapter 1.8 --- hexa-histidine tagged fusion protein --- p.25 / Chapter 1.9 --- IMAC --- p.26 / Chapter 1.10 --- Aim of study --- p.26 / Chapter 2 --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.29 / Chapter 2.2 --- Methods --- p.40 / Chapter 3 --- CONSTRUCTION AND CHARACTERIZATION OF RECOMBINANT BmNPV CARRYING PfMSP-l42 / Chapter 3.1 --- Cloning of C-HisPfMSP-l42 into pBM030 --- p.71 / Chapter 3.2 --- Construction of Recombinant BmNPV Carrying PfMSP-l42 --- p.72 / Chapter 3.3 --- Purification of Recombinant BmNPVs --- p.78 / Chapter 3.4 --- In vitro expression of Recombinant --- p.80 / Chapter 3.5 --- In Vivo Expression of Recombinant PfMSP-l42 Protein --- p.80 / Chapter 4 --- PURIFICATION OF BmNPV-EXPRESSED RECOMBINANT C- TERMIAL HEXA-HIS-TAGGED PfMSP-l42 PROTEIN / Chapter 4.1 --- Nickel ion charged Chelating Sepharose Fast Flow (immobilized metal affinity chromatography) --- p.88 / Chapter 4.2 --- POROS HS/M (Strong Cation Exchanger) --- p.105 / Chapter 4.3 --- Combination of chromatographic separations --- p.107 / Chapter 5 --- CHARACTERIZATION OF RECOMBINANT C-HISPfMSP-l42 PROTEIN / Chapter 5.1 --- Proper formation of disulphide bridges in epidermal growth factor (EGF) like domains --- p.115 / Chapter 5.2 --- Characterization of the integrity of hexa-histidines residue on recombinant PfMSP-142 protein --- p.117 / Chapter 5.3 --- Immunogenicity of Recombinant C-HisPfMSP-l42 Protein --- p.117 / Chapter 6 --- DISCUSSION / Chapter 6.1 --- Construction of recombinant BmNPV carrying HisPfMSP-l42 --- p.122 / Chapter 6.2 --- Expression of recombinant HisPfMSP-l42 proteins --- p.123 / Chapter 6.3 --- Purification of recombinant C-HisPfMSP-l42 protein --- p.125 / Chapter 6.4 --- Characterization of recombinant C-HisPfMSP-l42 protein --- p.128 / Chapter 6.5 --- Future prospects --- p.130 / REFERENCE --- p.135 / APPENDICES / Chapter 1. --- Appearance of Mulberry leaves / Chapter 2. --- Biomark 2000 (Beckman) program for sandwich ELISA protocol / Chapter 3. --- Nucleotide Sequence of PfMSP-l42 3D7 Isolate / Chapter 4. --- Nucleotide sequence of PfMSP-l42 FVO isolate / Chapter 5. --- Efficiency of the mAb5.2 immunoaffinity column in purifying the recombinant PfMSP-l42 protein

Liu Hua. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 139-143). / Abstracts in English and Chinese. / Table of Contents --- p.I / Acknowledgement --- p.VII / Abstract --- p.VIII / Abstract (Chinese) --- p.X / List of Tables --- p.XV / List of Figures --- p.XXIV / List Symbols and Abbreviations / Chapter Chapter One --- Introduction / Chapter 1.1. --- Rationale of study --- p.2 / Chapter 1.2. --- Drug Delivery to the lungs --- p.5 / Chapter 1.3. --- Particle transport and deposition mechanisms --- p.8 / Chapter 1.4. --- Factors affecting particulate interactions --- p.9 / Chapter 1.4.1. --- Particle size --- p.9 / Chapter 1.4.2. --- Particle shape --- p.10 / Chapter 1.4.3. --- Surface texture --- p.10 / Chapter 1.4.4. --- Surface energy --- p.11 / Chapter 1.4.5. --- Contact area --- p.12 / Chapter 1.4.6. --- Relative humidity --- p.12 / Chapter 1.4.7. --- Electrical properties --- p.13 / Chapter 1.5. --- Fine powder production technologies applicable to dry powder inhalation formulations --- p.13 / Chapter 1.5.1. --- Batch crystallization and micronization --- p.14 / Chapter 1.5.2. --- Spray drying --- p.15 / Chapter 1.5.3. --- Supercritical fluid crystallization --- p.17 / Chapter 1.6. --- Physical characterization of aerosol powders --- p.18 / Chapter 1.6.1. --- Microscopy and particle size analysis --- p.19 / Chapter 1.6.2. --- Powder X-ray diffractometry --- p.19 / Chapter 1.6.3. --- Thermal analysis --- p.20 / Chapter 1.6.4. --- In-vitro deposition assessment --- p.20 / Chapter 1.6.5. --- Surface energy measurement by inverse gas chromatography (IGC) --- p.21 / Chapter 1.7. --- Scope of study --- p.22 / Chapter Chapter Two --- Materials and Methods / Chapter 2.1. --- Materials --- p.25 / Chapter 2.2. --- Equipment --- p.25 / Chapter 2.3. --- Methods --- p.27 / Chapter 2.3.1. --- Determination of aqueous solubility of salbutamol sulphate in water --- p.27 / Chapter 2.3.2. --- Preparation of spray-dried salbutamol sulphate powders under different operating conditions --- p.27 / Chapter 2.3.3. --- Preparation of spray-dried salbutamol sulphate powders at various lecithin concentrations --- p.30 / Chapter 2.3.4. --- Preparation of spray-dried salbutamol sulphate powders at various oleic acid concentrations --- p.32 / Chapter 2.3.5. --- Physical characterization of spray-dried salbutamol sulphate powders --- p.34 / Chapter 2.3.5.1. --- Scanning electron microscopy --- p.34 / Chapter 2.3.5.2. --- Specific surface area determination --- p.34 / Chapter 2.3.5.3. --- Particle size distribution measurements --- p.35 / Chapter 2.3.5.4. --- Water content determination --- p.36 / Chapter 2.3.5.5. --- Isothermal water vapour sorption studies --- p.37 / Chapter 2.3.5.6. --- Powder X-ray diffraction --- p.37 / Chapter 2.3.5.7. --- Thermal analysis --- p.38 / Chapter 2.3.5.8. --- Surface energy measurement by inverse gas chromatography --- p.39 / Chapter 2.3.5.8.1. --- Calculation of surface thermodynamic parameters --- p.40 / Chapter 2.3.5.8.1.1. --- Standard Free Energy of Adsorption and Related Thermodynamic Parameters --- p.40 / Chapter 2.3.5.8.1.2. --- Dispersive Component of Surface Free Energy and Related Thermodynamic Parameters --- p.41 / Chapter 2.3.5.8.1.3. --- Specific Interactions and Associated Acid-Base Properties --- p.42 / Chapter 2.3.5.9. --- In-vitro deposition measurement by multi-stage liquid impinger --- p.43 / Chapter Chapter Three --- Results and discussion / Chapter 3.1. --- Influence of spray drying operating parameters --- p.46 / Chapter 3.1.1. --- Drying temperature --- p.46 / Chapter 3.1.1.1. --- "Particle size, particle morphology, and specific surface area" --- p.46 / Chapter 3.1.1.2. --- "Crystallinity, moisture sorption and thermal behaviour" --- p.53 / Chapter 3.1.1.3. --- Surface thermodynamic properties --- p.60 / Chapter 3.1.1.4. --- Aerodynamic properties and in-vitro deposition --- p.64 / Chapter 3.1.2. --- Feed solution concentration --- p.67 / Chapter 3.1.2.1. --- "Particle size, particle morphology and specific surface area" --- p.69 / Chapter 3.1.2.2. --- "Crystallinity, moisture sorption and thermal behaviour" --- p.69 / Chapter 3.1.2.3. --- Surfacethermodynamicproperties --- p.70 / Chapter 3.1.2.4. --- Aerodynamic properties and in-vitro deposition --- p.70 / Chapter 3.1.3. --- Feed speed --- p.72 / Chapter 3.1.3.1. --- "Particle size, particle morphology, and specific surface area" --- p.72 / Chapter 3.1.3.2. --- "Crystallinity, moisture sorption and thermal behaviour" --- p.73 / Chapter 3.1.3.3. --- Surfacethermodynamicproperties --- p.73 / Chapter 3.1.3.4. --- Aerodynamic properties and in-vitro deposition --- p.73 / Chapter 3.2. --- Influence of formulation additives --- p.78 / Chapter 3.2.1. --- Influence of lecithin as additive --- p.78 / Chapter 3.2.1.1. --- "Particle morphology, particle size and specific surface area" --- p.79 / Chapter 3.2.1.2. --- "Crystallinity, moisture sorption and thermal behaviour" --- p.84 / Chapter 3.2.1.3. --- Surfacethermodynamicproperties --- p.90 / Chapter 3.2.1.4. --- Aerodynamic properties and in-vitro deposition --- p.94 / Chapter 3.2.2. --- Influence of oleic acid as additive --- p.101 / Chapter 3.2.2.1. --- "Particle morphology, particle size and specific surface area" --- p.101 / Chapter 3.2.2.2. --- "Crystallinity, moisture sorption and thermal behaviour" --- p.106 / Chapter 3.2.2.3. --- Surfacethermodynamicproperties --- p.123 / Chapter 3.2.2.4. --- Aerodynamic properties and in-vitro deposition --- p.127 / Chapter Chapter Four --- Conclusion and Future Work / Chapter 4.1. --- Conclusion --- p.134 / Chapter 4.1.1. --- Influence of spray drying operating parameters --- p.134 / Chapter 4.1.2. --- Influence of formulation additives --- p.135 / Chapter 4.2. --- Future Work --- p.137 / References --- p.139

by Leung Ho-yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references. / Abstract also in Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / Table of Contents --- p.iv / List of Tables and Figures --- p.v / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Methodology --- p.12 / Chapter Chapter 3 --- Experimental --- p.17 / Chapter 3.1 --- Sample Collection --- p.17 / Chapter 3.2 --- Preliminary Sample Treatment --- p.18 / Chapter 3.3 --- Dissolution Test Apparatus --- p.18 / Chapter 3.4 --- Reagents --- p.20 / Chapter 3.5 --- ICP Experimental Conditions --- p.21 / Chapter Chapter 4 --- Determination of Total Contents in Chinese Mineral Drugs --- p.28 / Chapter 4.1 --- Procedure --- p.28 / Chapter 4.2 --- Results --- p.39 / Chapter Chapter 5 --- Determination of the Effective Contents --- p.51 / Chapter 5.1 --- General Procedure --- p.51 / Chapter 5.2 --- Results --- p.52 / Chapter Chapter 6 --- Recovery Test for Dissolution --- p.77 / Chapter 6.1 --- General Procedure --- p.77 / Chapter 6.2 --- Results --- p.78 / Chapter Chapter 7 --- Discussion --- p.80 / Chapter 7.1 --- The Presence of Toxic Elements in many CMD --- p.80 / Chapter 7.2 --- Mass of Samples Added in the Dissolution --- p.80 / Chapter 7.3 --- Height of the Dissolution Paddle of the Dissolution --- p.81 / Chapter 7.4 --- Particle Sizes of the Sample in the Dissolution --- p.83 / Chapter 7.5 --- The Total and the Effective Contents --- p.83 / Chapter 7.6 --- Discussion for each CMD --- p.84 / Chapter 7.7 --- Conclusion --- p.103 / Appendix English 一 Chinese table of CMD --- p.108

隨著社會發展, 各種病毒、環境致癌物, 以及不健康食等多種因素導致諸種頑症高發, 並以人類獲得牲免疫缺陷綜合症(艾滋病)和各種腫瘤為代表。從天然產物, 特別是傳統中藥中, 篩選藥用有效成分是治療這類疾病的有效途徑之一。研究發現, 核糖體失活蛋白, 核糖核酸酶, 凝集素, 蛋白酪抑制劑等具有良好的藥用開發前景。本論文著重於從不同植物種于中篩選藥用蛋白, 並對其藥用機制進行研究。 / 是吹研究共純化出六種藥用蛋白。第一, 從苦瓜種子中純化出一種二型核糖體失活蛋白MCL。體外細胞試驗和體內裸鼠試驗顯示MCL 能夠有效抑制鼻咽癌細胞CNE-l 和CNE-2 生長。第二, 苦瓜種于中一個新的核糖核酸酪RNase MC2被分離出來。RNase MC2 通過調控半胱氨酸依賴性細胞死亡蛋白晦(Caspase)信號途徑和絲裂原活化蛋白激酶(MAPKs) 信號途徑誘導乳腺癌MCF-7 細胞凋亡。第三, 從宮粉羊蹄甲種子中提取一種具有抑制腫瘤細胞生長的蛋白酪抑制劑BvvTI 。第四, 從紅花羊蹄甲種子中提取出一與BvvTI 類似的具有抗腫瘤生長的蛋白酶抑制劑BPLTI。 第五，特長秋紫莢豆中存在一個血液凝集素EAPl。EAPL具有制止HIV-l 逆轉錄酶活性, 抗腫瘤, 誘導一氧化氮生成功效。第六, 從另外一種四季豆, 藍虎王, 中提取出一個血液凝集素BTKL。BTKL 通過誘導肝癌HepG2細胞出現DNA 斷裂, 細胞核破壞, 升高線粒體膜通透性, 誘導一氧化氮和細胞因予的表達, 從而導致其凋亡。 / 總之, 上述實驗結果表明, 這六種蛋白具有一定的藥用前景, 可以作為治療艾滋病和不同腫瘤的候造藥物或者候選輔助藥物。進一步體內實驗和臨床實驗評價其療效值得開展。 / Viral pathogens, environmental carcinogens, and unhealthy diets cause severe damage to humans, leading to the acquirement of different stubborn diseases exemplified by AIDS/HIV and neoplasms. Screening of new drugs from natural products, especially from traditional Chinese medicine, provides a promising strategy for these patients. Proteins with potential medicinal applications include ribosome-inactivating proteins (RIPs), ribonucleases, lectins, protease inhibitors and others. The intent of this research proposal is to isolate and characterize proteins with therapeutic potential from plant seeds. / In this project, six medicinal proteins of different origins have been purified by liquid chromatography. One of them is Momordica charantia lectin (MCL), which is a type 11 RIP from the seeds of bitter gourd (M. charantia, BG), with antitumor activity toward human nasopharyngeal carcinoma cells in vitro and in vivo. We have purified a new ribonuclease, named RNase MC2, from BG seeds. It selectively induces apoptosis in breast cancer cells associated with caspase pathways induction and MAPKs activation. Two Kunitz-type trypsin inhibitors, termed BvvTI and BPLTI, have been purified and characterized from the seeds of Bauhinia variegata var. variegata, and B. purpurea L., respectively. EAPL, a lectin with anti-HIV-l reverse transcriptase, antitumor, and nitric oxide (NO) inducing activities was purified from seeds of Phaseolus vulgaris cv. Extralong autumn purple bean. Finally, BTKL is a new P. vulgaris lectin that induced selective toxicity on human liver carcinoma Hep G2 cells. / In conclusion, the above results evince that these proteins are good candidates for the exploration of anti-HIV and/or antitumor drugs or adjuvants. Further research on their efficacies in in vivo as well as clinical trials is warranted. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Fang, Fei. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 162-187). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / ABSTRACT --- p.ii / 中文摘要 (CHINESE ABSTRACT) --- p.iii / ACKNOWLEDGEMENTS --- p.iv / PUBLICATIONS --- p.v / LIST OF ABBREViATIONS --- p.ix / LIST OF FIGURES --- p.x / LIST OF TABLES --- p.xii / Chapter CHAPTER1 --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Prelude --- p.2 / Chapter 1.2 --- Literature review of bitter gourd --- p.4 / Chapter 1.2.1 --- Anti-diabetic property of BG --- p.5 / Chapter 1.2.2 --- Anti-HIV activity of BG --- p.15 / Chapter 1.2.3 --- Antitumor activity of BG --- p.18 / Chapter 1.2.4 --- Looking forward --- p.25 / Chapter 1.2.5 --- Conclusions --- p.29 / Chapter 1.3 --- Research rationale, design, and brief results --- p.35 / Chapter CHAPTER2 --- PURIFICATION AND CHARACTERIZATION OF RIBOSOME INACTIVATING PROTEIN --- p.39 / Chapter 2.1 --- Momordica charantia lectin, a type 11 ribosome inactivating protein, exhibits antitumor activity toward human nasopharyngeal carcinoma cells in vitro and in vivo --- p.40 / Chapter 2.1.1 --- Introduction --- p.40 / Chapter 2.1.2 --- Materials and methods --- p.42 / Chapter 2.1.3 --- Results --- p.46 / Chapter 2.1.4 --- Discussion --- p.59 / Chapter CHAPTER 3 --- PURIFICATION AND CHARACTERIZATION OF RIBONUCLEASE --- p.64 / Chapter 3.1 --- RNase MC2: A new Momordica charantia ribonuclease that selectively induces apoptosis in breast cancer cells associated with MAPKs activation and caspase pathways induction --- p.66 / Chapter 3.1.1 --- Introduction --- p.66 / Chapter 3.1.2 --- Materials and methods --- p.67 / Chapter 3.1.3 --- Results --- p.69 / Chapter 3.1.4 --- Discussion --- p.79 / Chapter CHAPTER 4 --- PURIFICATION AND CHARACTERIZATION OF PROTEASE INHIBITORS --- p.84 / Chapter 4.1 --- Bauhinia variegata var variegata trypsin inhibitor: from isolation to potential medicinal applications --- p.86 / Chapter 4.1.1 --- Introduction --- p.86 / Chapter 4.1.2 --- Materials and methods --- p.86 / Chapter 4.1.3 --- Results --- p.89 / Chapter 4.1.4 --- Discussion --- p.97 / Chapter 4.2 --- A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L.seeds: From purification to mechanism exploration --- p.99 / Chapter 4.2.1 --- Introduction --- p.99 / Chapter 4.2.2 --- Materials and methods --- p.100 / Chapter 4.2.3 --- Results --- p.101 / Chapter 4.2.4 --- Discussion --- p.112 / Chapter CHAPTER 5 --- PURIFICATION AND CHARACTERIZATION OF MEDICINAL LECTINS --- p.114 / Chapter 5.1 --- A Lectin with Anti-HIV-l Reverse Transcriptase, Antitumor and Nitric Oxide Inducing Activities from Seeds of Phaseolus vulgaris cv Extra-long Autumn Purple Bean --- p.116 / Chapter 5.1.1 --- Introduction --- p.118 / Chapter 5.1.2 --- Materials and methods --- p.119 / Chapter 5.1.3 --- Results --- p.123 / Chapter 5.1.4 --- Discussion --- p.132 / Chapter 5.2 --- A new Phaseolus vulgaris lectin induces selective toxicity on human liver carcinoma Hep G2 cells --- p.136 / Chapter 5.2.1 --- Introduction --- p.136 / Chapter 5.2.2 --- Materials and methods --- p.137 / Chapter 5.2.3 --- Results --- p.138 / Chapter 5.2.4 --- Discussion --- p.150 / Chapter CHAPTER6 --- CONCLUSION AND FUTURE PERSPECTiVES --- p.154 / Chapter 6.1 --- Conclusions --- p.156 / Chapter 6.2 --- Future perspectives --- p.159 / References --- p.162

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