Chan Ching-Man Loren. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 126-144). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / Table of Content --- p.vii / List of Figure --- p.ix / List of Abbreviations --- p.xii / Chapter Chapter One --- General Introduction / Chapter 1.1 --- Doxorubicin --- p.1 / Chapter 1.2 --- Antioxidants --- p.11 / Chapter 1.3 --- Flavonoids --- p.17 / Chapter 1.4 --- Protection against doxorubicin-induced cardiotoxicity by antioxidant --- p.25 / Chapter 1.5 --- Aim of research --- p.25 / Chapter Chapter Two --- Study of Cardioprotection Effect of Flavonoids Against Doxorubicin-induced Toxicity in Sprague-Dawley Rats / Chapter 2.1 --- Introduction --- p.27 / Chapter 2.2 --- Materials and Methods --- p.29 / Chapter 2.3 --- Results --- p.37 / Chapter 2.4 --- Discussion --- p.51 / Chapter ChapterThree --- Study of Effect of Flavonoids in Chemotherapy of Doxorubicinin Tumor-bearing Mice / Chapter 3.1 --- Introduction --- p.54 / Chapter 3.2 --- Materials and Methods --- p.56 / Chapter 3.3 --- Results --- p.63 / Chapter 3.4 --- Discussion --- p.80 / Chapter ChapterFour --- Study of the Effect of Flavonoids on Doxorubicin-induced Cytotoxicity on Human Tumor Cell Line and Doxorubicin-resistant Human Tumor Cell Line / Chapter 4.1 --- Introduction --- p.83 / Chapter 4.2 --- Materials and Methods --- p.86 / Chapter 4.3 --- Results --- p.94 / Chapter 4.4 --- Discussion --- p.120 / Chapter Chapter five --- Conclusion --- p.122 / References --- p.126

by Kong Lai Ping, Ada. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 171-189). / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.v / TABLE OF CONTENTS --- p.ix / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- An Overview on Hematopoiesis --- p.1 / Chapter 1.1.1 --- Development of Hematopoietic Stem Cells and Sites of Hematopoiesis --- p.1 / Chapter 1.1.2 --- Role of Cytokines in the Control of Hematopoiesis --- p.3 / Chapter 1.2 --- Leukemia and Cell Differentiation --- p.5 / Chapter 1.2.1 --- Leukemia as Abnormalities in Hematopoietic Cell Development --- p.5 / Chapter 1.2.2 --- Classification and Etiology of Leukemia --- p.6 / Chapter 1.2.3 --- Current Modalities for the Treatment of Leukemia --- p.9 / Chapter 1.2.4 --- Leukemia Cell Lines as In Vitro Models for the Study of Myeloid Leukemia --- p.10 / Chapter 1.2.5 --- Cytokines as Inducers of Myeloid Leukemia Cell Differentiation --- p.12 / Chapter 1.2.6 --- The Murine Myeloid Leukemia Cell Line (WEHI- 3B JCS) as an Experimental Cell Model --- p.13 / Chapter 1.3 --- Flavonoids: Properties and Biological Activities --- p.15 / Chapter 1.3.1 --- Chemical Structure and Classification of Flavonoids --- p.15 / Chapter 1.3.2 --- Occurrence and Distribution of Flavonoids --- p.16 / Chapter 1.3.3 --- Biological Properties and Action Mechanisms of Flavonoids --- p.17 / Chapter 1.3.4 --- Effects of Flavonoids on Leukemia --- p.20 / Chapter 1.4 --- Aims and Scopes of This Investigation --- p.23 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.26 / Chapter 2.1.1 --- Cell Lines --- p.26 / Chapter 2.1.2. --- Mice --- p.28 / Chapter 2.1.3 --- Flavonoids --- p.28 / Chapter 2.1.4 --- Recombinant Cytokines --- p.30 / Chapter 2.1.5. --- Physiological Differentiation Inducers ´ؤ Vitamin Analogs --- p.31 / Chapter 2.1.6 --- Monoclonal Antibodies --- p.31 / Chapter 2.1.7 --- "Buffers, Culture Medium and Other Reagents" --- p.33 / Chapter 2.1.8 --- Oligonucleotide Primers and Internal Probes --- p.36 / Chapter 2.1.9 --- Reagents for Cytokine Gene Expression Study --- p.38 / Chapter 2.2 --- Methods --- p.44 / Chapter 2.2.1 --- Culture of Tumor Cell Lines --- p.44 / Chapter 2.2.2 --- Determination of Cell Growth and Proliferation --- p.45 / Chapter 2.2.3 --- Colony Assay --- p.46 / Chapter 2.2.4 --- In vivo Tumorigenicity Assay --- p.46 / Chapter 2.2.5 --- Induction of Leukemic Cell Differentiation --- p.47 / Chapter 2.2.6 --- Cell Morphological Study --- p.47 / Chapter 2.2.7 --- Assessment of Differentiation Associated Characteristics --- p.48 / Chapter 2.2.7.1 --- Nitroblue Tetrazolium (NBT) Reduction Assay --- p.48 / Chapter 2.2.7.2 --- Assay of Plastic Adherence --- p.48 / Chapter 2.2.8 --- Flow Cytometric Analysis --- p.49 / Chapter 2.2.8.1 --- Surface Antigen Immunophenotyping --- p.49 / Chapter 2.2.8.2 --- Assay of Non-specific Esterase Activity --- p.50 / Chapter 2.2.8.3 --- Assay of Phagocytic Activity --- p.50 / Chapter 2.2.8.4 --- Assay of Endocytic Activity --- p.51 / Chapter 2.2.8.5 --- Cell Cycle/DNA Content Evaluation --- p.52 / Chapter 2.2.9 --- Gene Expression Analysis --- p.53 / Chapter 2.2.9.1 --- Cell Lysate Preparation --- p.53 / Chapter 2.2.9.2 --- Total RNA Isolation by cesium chloride isopycnic gradient --- p.53 / Chapter 2.2.9.3 --- Reverse Transcription --- p.54 / Chapter 2.2.9.4 --- Polymerase Chain Reaction (PCR) --- p.55 / Chapter 2.2.9.5 --- Agarose Gel Electrophoresis --- p.56 / Chapter 2.2.9.6 --- DIG 3，End Labeling of Oligonucleotide Probes --- p.57 / Chapter 2.2.9.7 --- Dot Blot Hybridization --- p.57 / Chapter 2.2.9.8 --- DIG Chemiluminescent Detection --- p.58 / Chapter 2.2.10 --- DNA Fragmentation Analysis --- p.59 / Chapter 2.2.11 --- Statistical Analysis --- p.60 / Chapter CHAPTER 3: --- EFFECTS OF FLAVONOIDS ON THE PROLIFERATION AND APOPTOSIS OF MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.2 --- Results --- p.63 / Chapter 3.2.1 --- Growth-Inhibitory Effects of Flavone on Murine Myeloid Leukemia JCS Cells --- p.63 / Chapter 3.2.2 --- Cytotoxic Effects of Flavone on Murine Lymphocytes and Myeloid Leukemia JCS Cells --- p.67 / Chapter 3.2.3 --- Effects of Different Flavonoids on the Proliferation of Leukemia JCS Cells --- p.70 / Chapter 3.2.4 --- Anti-proliferative Effect of Flavonoids on Different Tumor Cell Lines --- p.74 / Chapter 3.2.5 --- Effects of Flavone and Flavonol on the Cell Cycle Kinetics of JCS Cells --- p.86 / Chapter 3.2.6 --- Induction of DNA Fragmentation of JCS cells by Flavone --- p.89 / Chapter 3.2.7 --- Effect of Flavone on the Clonogenicity of JCS Cells In Vitro and Tumorigenicity In Vivo --- p.92 / Chapter 3.3 --- Discussion --- p.94 / Chapter CHAPTER 4: --- EFFECTS OF FLAVONOIDS ON THE DIFFERENTIATION OF MURINE MYELOID LEUKEMIA JCS CELLS / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Results --- p.100 / Chapter 4.2.1 --- Morphological Changes in Flavonoid-Treated JCS Cells --- p.100 / Chapter 4.2.2 --- Induction of Plastic Adherence in Flavonoid- Treated JCS Cells --- p.106 / Chapter 4.2.3 --- Surface Antigen Immunophenotyping of Differentiating JCS Cells --- p.106 / Chapter 4.2.4 --- NBT-Reducing Activity of Flavonoid-Treated JCS Cells --- p.114 / Chapter 4.2.5 --- Non-specific Esterase Activity of Flavonoid- Treated JCS Cells --- p.115 / Chapter 4.2.6 --- Endocytic Activity of Flavonoid-Treated JCS Cells --- p.116 / Chapter 4.2.7 --- Phagocytic Activity of Flavonoid-Treated JCS Cells --- p.117 / Chapter 4.3 --- Discussion --- p.118 / Chapter CHAPTER 5: --- MECHANISTIC STUDIES ON THE ANTI- PROLIFERATIVE AND DIFFERENTIAION-INDUCING ACTIVITIES OF FLAVONE ON MURINE MYELOID LEUKEMIA JCS CELLS / Chapter 5.1 --- Introduction --- p.122 / Chapter 5.2 --- Results --- p.125 / Chapter 5.2.1 --- Combinations of Flavone with Physiological Differentiation Inducers on the Proliferation and Differentiation of JCS Cells --- p.125 / Chapter 5.2.1.1 --- Modulatory Effects of Flavone and All-Trans Retinoic Acid (ATRA) on the Proliferation and Differentiation of JCS Cells --- p.125 / Chapter 5.2.1.2 --- "Modulatory Effects of Flavone and 1,25- dihydroxyvitamin D3 on the Proliferation and Differentiation of JCS Cells" --- p.130 / Chapter 5.2.2 --- Combinations of Flavone and Cytokines on the Proliferation and Differentiation of JCS Cells --- p.134 / Chapter 5.2.2.1 --- Modulatory Effects of Flavone and rmlFN-γ on the Proliferation and Differentiation of JCS Cells --- p.134 / Chapter 5.2.2.2 --- Synergistic Effects of Flavone and rmIL-1 on the Proliferation and Differentiation of JCS Cells --- p.137 / Chapter 5.2.3 --- Modulation of Cytokine Gene Expressionin Flavone-Treated JCS Cells --- p.144 / Chapter 5.3 --- Discussion --- p.159 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.165 / REFERENCES --- p.171

Chan Ho Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 121-138). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.II / ABSTRACT --- p.III / 摘要 --- p.V / ABBREVIATIONS --- p.VI / "TABLE OF CONTENTS, " --- p.VII / LIST OF FIGURES AND TABLES --- p.XI / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Xenobiotic-metabolizing enzymes --- p.1 / Cytochrome P450 1 family --- p.4 / CYP1A1 --- p.5 / CYP1A2 --- p.5 / CYP1B1 --- p.5 / Transactivation of CYP1 enzymes by Aryl hydrocarbon receptor (AhR) --- p.8 / Implication of PAHs and CYP1 family in breast cancer --- p.10 / Potential role of phytochemicals on cancer prevention --- p.11 / Significance of this project --- p.13 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.14 / Chemicals --- p.14 / Maintenance of cells --- p.14 / Preparation of cell stock --- p.14 / Cell recovery from liquid nitrogen stock --- p.15 / Measurement of cell viability --- p.15 / Preparation of cell lysates (NP-40 cell lysis buffer) --- p.15 / XRE-luciferase gene reporter assay --- p.16 / Manipulation of DNA and RNA --- p.17 / Separation and purification of DNA from agarose gel --- p.17 / Separation of DNA from acrylamide gel --- p.17 / Restriction digestion --- p.18 / Ligation of DNA fragments --- p.18 / Transformation of DH5 a --- p.19 / Small scale plasmid purification from DH5a (mini prep) --- p.19 / Large scale plasmid isolation from DH5a (maxi-prep) --- p.20 / Construction of XRE activated luciferase reporter gene --- p.21 / Measurement of DMBA-DNA adduct formation --- p.21 / Semi-quantitative RT-PCR Assay --- p.22 / ENZYME ACTIVITIES --- p.23 / Isolation of microsomes --- p.23 / EROD activities in intact cells --- p.23 / EROD inhibition assay --- p.24 / Stattstical Analysis --- p.24 / Chapter CHAPTER 3 --- BAICALEIN INHIBITS DMBA-DNA ADDUCT FORMATION BY MODULATING CYP1A1 AND 1B1 ACTIVITIES --- p.26 / Introduction --- p.26 / Results --- p.28 / EROD activities in MCF-7 cells and inhibition assay --- p.28 / Baicalein suppressed DMBA-induced XRE-driven luciferase activities --- p.31 / Baicalein inhibited DMBA-induced CYP1A1 and CYP1B1 mRNA expression --- p.31 / The cytotoxic effect of DMBA was reduced by baicalein --- p.35 / Inhibition of DMBA-DNA adduct formation after baicalein treatment --- p.35 / Discussion --- p.39 / Chapter CHAPTER 4 --- INHIBITION OF DMBA-DNA ADDUCT FORMATION BY (-)-EPIGALLOCATECHIN GALLATE AND THEAFLAVINS --- p.41 / Introduction --- p.41 / Results --- p.45 / Persistence of DMBA-induced DNA adducts --- p.45 / Inhibition of theaflavins and EGCG on human recombinant CYP1A1 and CYP1B1 enzyme activities --- p.48 / EGCG suppressed DMBA-induced EROD activity while thealfavin had no significant effect on this --- p.48 / Kinetic analysis of EGCG on CYP1A1 and CYP1B1 activities --- p.53 / Modulation of DMBA-induced XRE-driven luciferase activities by theaflavins and EGCG --- p.56 / The influence of theaflavins and EGCG on CYP1A1 and CYP1B1 abundance --- p.56 / Discussion --- p.65 / Chapter CHAPTER 5 --- ISOFLAVONES PREVENT DMBA-INDUCED CARCINOGENESIS BY INHIBITING CYP1A1 AND CYP1B1 ACTIVITIES --- p.67 / Introduction --- p.67 / Results --- p.70 / Isoflavones inhibited DMBA-induced EROD activity in MCF-7 cells --- p.70 / Inhibition of MCF-7 microsomal EROD activities by isoflavones --- p.70 / Kinetic analysis of the inhibition of human recombinant CYP1 enzymes by isoflavones --- p.74 / XRE-driven Luciferase activities --- p.83 / Both biochanin A and genistein suppressed DMBA-induced CYP1 mRNA expression --- p.83 / Cytotoxicity of DMBA and isoflavones co-treatment --- p.88 / Isoflavones reduced the binding of activated DMBA to DNA --- p.89 / Discussion --- p.93 / Chapter CHAPTER 6 --- IN VITRO EFFECTS OF BAICALEIN AND THEAFLAVINS ON RAT HEPATIC P450 ACTIVITIES --- p.96 / Introduction --- p.96 / Results --- p.98 / Inhibition of EROD and MROD activities in rat liver microsomes by baicalein --- p.98 / Effects of theaflavins on EROD and MROD activities in rat liver microsomes --- p.102 / Kinetic studies for EROD and MROD activities of theaflavins --- p.104 / DISCUSSION --- p.114 / Chapter CHAPTER 7 --- CONCLUSION --- p.116 / APPENDIX 1 PRIMER LISTS --- p.118 / APPENDIX 2 REAGENTS --- p.119 / BIBLIOGRAPHY --- p.121