Leung Pui-Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 226-266). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.iii / ABSTRACT --- p.viii / 撮要 --- p.xii / TABLE OF CONTENTS --- p.xvi / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis & Leukemia --- p.1 / Chapter 1.1.1 --- Introduction to Hematopoiesis and its Regulation --- p.1 / Chapter 1.1.2 --- Leukemia --- p.7 / Chapter 1.1.2.1 --- Classification and Epidemiology of Leukemia --- p.7 / Chapter 1.1.2.2 --- Conventional Approaches to Leukemia Therapy --- p.13 / Chapter 1.1.2.3 --- New Approaches to Leukemia Therapy --- p.16 / Chapter 1.2 --- Coumarins --- p.18 / Chapter 1.2.1 --- Introduction to Coumarins --- p.18 / Chapter 1.2.1.1 --- "Historical Development, Occurrence of Coumarins and their Functions in Plants" --- p.18 / Chapter 1.2.1.2 --- Beneficial Effects of Coumarins for Human Use --- p.20 / Chapter 1.2.2 --- Phytochemistry and Metabolism of Coumarins --- p.22 / Chapter 1.2.2.1 --- Chemical Structures of Coumarins --- p.22 / Chapter 1.2.2.2 --- Biosynthesis of Coumarins --- p.24 / Chapter 1.2.2.3 --- Toxicology of Coumarins --- p.24 / Chapter 1.2.2.4 --- Mode of Entry of Coumarins --- p.25 / Chapter 1.2.2.5 --- Metabolic Pathways of Coumarins --- p.27 / Chapter 1.2.3 --- Pharmacological Activities of Coumarins --- p.30 / Chapter 1.2.3.1 --- Anti-edema and Anti-inflammatory Activities --- p.30 / Chapter 1.2.3.2 --- Cardiovascular Protective Function --- p.32 / Chapter 1.2.3.3 --- Anti-tumor Activity --- p.33 / Chapter 1.2.3.3.1 --- Anti-tumor Activity In Vitro --- p.33 / Chapter 1.2.3.3.2 --- Anti-tumor Activity In Vivo and Clinical Applications --- p.34 / Chapter 1.2.3.4 --- Immunomodulatory Activity --- p.36 / Chapter 1.2.3.4.1 --- Immunological Considerations --- p.36 / Chapter 1.2.3.4.2 --- Immunomodulation In Vitro --- p.37 / Chapter 1.2.3.4.3 --- Immunomodulation In Vivo --- p.38 / Chapter 1.3 --- Aims and Scopes of This Investigation --- p.41 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.43 / Chapter 2.1.1 --- Animals --- p.43 / Chapter 2.1.2 --- Cell Lines --- p.43 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.44 / Chapter 2.1.4 --- Reagents for 3H-Thymidine Incorporation Assay --- p.49 / Chapter 2.1.5 --- Reagents and Buffers for Flow Cytometry --- p.49 / Chapter 2.1.6 --- Reagents for DNA Extraction --- p.52 / Chapter 2.1.7 --- Cell Death Detection ELISAplus Kit --- p.54 / Chapter 2.1.8 --- Reagents for Total RNA Isolation --- p.55 / Chapter 2.1.9 --- Reagents and Buffers for RT-PCR --- p.56 / Chapter 2.1.10 --- Reagents and Buffers for Gel Electrophoresis of Nucleic Aicds --- p.60 / Chapter 2.1.11 --- "Reagents, Buffers and Materials for Western Blot Analysis" --- p.61 / Chapter 2.1.12 --- Reagents for Measuring Caspase Activity --- p.67 / Chapter 2.1.13 --- Reagents for Neutral Red Assay --- p.70 / Chapter 2.2 --- Methods --- p.71 / Chapter 2.2.1 --- Culture of the Tumor Cell Lines --- p.11 / Chapter 2.2.2 --- Determination of Cell Proliferation by [3H]-TdR Incorporation Assay --- p.71 / Chapter 2.2.3 --- Determination of Cell Viability --- p.72 / Chapter 2.2.4 --- In Vivo Anti-tumor Study --- p.73 / Chapter 2.2.5 --- Analysis of Cell Cycle Profile/DNA Content by Flow Cytometry --- p.74 / Chapter 2.2.6 --- Measurement of Apoptosis --- p.74 / Chapter 2.2.7 --- "Islation ,Preparation and Culture of Mouse Peritoneal Macrophages" --- p.76 / Chapter 2.2.8 --- Gene Expression Study --- p.77 / Chapter 2.2.9 --- Protein Expression Study --- p.80 / Chapter 2.2.10 --- Determination of the Mitochondrial Membrane Potential --- p.84 / Chapter 2.2.11 --- Measurement of Caspase Activity --- p.84 / Chapter 2.2.12 --- In Vivo Macrophage Migration Assay --- p.85 / Chapter 2.2.13 --- Study of Endocytic Activity of Macrophages --- p.85 / Chapter 2.2.14 --- Determination of Nitric Oxide Production by Macrophages --- p.86 / Chapter 2.2.15 --- Study of Mitogenesis of Murine Splenocytes --- p.87 / Chapter 2.2.16 --- Analysis of T Cell Population and its Sub-populations --- p.88 / Chapter 2.2.17 --- Measurement of Lymphokine Activated Killer (LAK) Cell Activity by Neutral Red Assay --- p.90 / Chapter 2.2.18 --- Statistical Analysis --- p.91 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI PROLIFERATIVE EFFECT OF COUMARINS ON MACROPHAGE-LIKE LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.92 / Chapter 3.2 --- Results --- p.94 / Chapter 3.2.1 --- Anti-proliferative Effect of Coumarins on Various Macrophage-like Leukemia Cell Lines In Vitro --- p.94 / Chapter 3.2.2 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Coumarins on Macrophage-like Leukemia PU5-1.8 Cells --- p.105 / Chapter 3.2.3 --- Cytotoxic Effect of Coumarins on Macrophage-like Leukemia PU5-1.8 Cells In Vitro --- p.110 / Chapter 3.2.4 --- Effect of Coumarins on the Cell Cycle Kinetics of the Macrophage-like Leukemia PU5-1.8 Cells In Vitro --- p.113 / Chapter 3.2.5 --- Effect of Coumarins on the Expression of Cell Cycle- and Growth-regulatory Genes in the Macrophage-like Leukemia PU5-1.8 Cells --- p.119 / Chapter 3.2.6 --- Effect of Coumarins on the Expression of Cell Cycle-regulatory Proteins in the Macrophage-like Leukemia PU5-1.8 Cells --- p.123 / Chapter 3.2.7 --- Effect of Coumarins on the In Vivo Tumorigenicity of the Macrophage-like Leukemia PU5-1.8 Cells --- p.127 / Chapter 3.2.8 --- Effect of Coumarins on the In Vivo Growth of the Macrophage- like Leukemia PU5-1.8 Cells in Syngeneic BALB/c Mice --- p.131 / Chapter 3.3 --- Discussion --- p.133 / Chapter CHAPTER 4: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF COUMARINS ON MACROPHAGE-LIKE LEUKEMIA PU5- CELLS --- p.1.8 / Chapter 4.1 --- Introduction --- p.143 / Chapter 4.2 --- Results --- p.152 / Chapter 4.2.1 --- Induction of Apoptosis in the Murine Macrophage-like Leukemia PU5-1.8 Cells by Coumarins --- p.152 / Chapter 4.2.2 --- Modulatory Effect of Coumarins on the Expression of Apoptosis-regulatory Genes in the Macrophage-like Leukemia PU5-1.8 Cells --- p.159 / Chapter 4.2.3 --- Modulatory Effect of Coumarins on the Expression of Apoptosis-regulatory Proteins in the Macrophage-like Leukemia PU5-1.8 Cells --- p.166 / Chapter 4.2.4 --- Effect of Coumarins on the Mitochondrial Membrane Depolarization of the Macrophage-like Leukemia PU5-1.8 Cells --- p.169 / Chapter 4.2.5 --- Effect of Coumarins on the Induction of Caspase Activity in the Macrophage-like Leukemia PU5-1.8 Cells --- p.172 / Chapter 4.2 --- Discussion --- p.177 / Chapter CHAPTER 5: --- STUDIES ON THE IMMUNOMODULATORY EFFECT OF COUMARINS ON MURINE MACROPHAGES AND OTHER IMMUNE CELLS / Chapter 5.1 --- Introduction --- p.184 / Chapter 5.2 --- Results --- p.187 / Chapter 5.2.1 --- Effect of Coumarins on the Viability of Macrophages In Vitro --- p.187 / Chapter 5.2.2 --- Effect of Coumarins on the In Vivo Migration of Macrophages --- p.190 / Chapter 5.2.3 --- Effect of Coumarins on the Endocytic Ability of Murine Macrophages --- p.192 / Chapter 5.2.4 --- Effect of Coumarins on the Nitric Oxide Production by Murine Macrophages --- p.194 / Chapter 5.2.5 --- Effect of Coumarins on the Viability of Murine Splenocytes --- p.199 / Chapter 5.2.6 --- Effect of Coumarins on the Mitogenesis of Murine Splenocytes --- p.201 / Chapter 5.2.7 --- Effect of Coumarins on T-cell Population and its Sub-populations --- p.205 / Chapter 5.2.8 --- Effect of Coumarins on the Induction of Murine LAK Activity --- p.208 / Chapter 5.3 --- Discussion --- p.210 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.218 / REFERENCES --- p.226