A cell surface protein from human acute myelogenous leukemia (AML) cells has been purified. (Al-Rammahy et al., Cancer Immunol. Immunother. 9:181, 1980; Malcolm et al., J. Immunol. 128:2599, 1982). This material was used to immunize rabbits. The resulting antiserum (anti-AML) showed myelogenous leukemia specificity in that it reacted with myelogenous leukemia cell extracts and did not react with cell extracts of normal individuals or patients with non-myelogenous leukemia or other malignant disorders in the enzyme-linked immunosorbent assay (ELISA). Bone marrow and peripheral blood leucocytes (PBL) from either patients with myelogenous leukemia, other disorders or normal individuals were analysed in the fluorescence-activated cell sorter (FACS IV) after labelling with anti-AML, normal rabbit serum (NRS), or antiserum raised to normal human membrane antigens. Of 40 cell samples from patients with AML, 39 reacted strongly with the anti-AML. Similarly, all of 15 specimens from patients with chronic myelogenous leukemia (CML) reacted with the anti-AML. When 42 bone marrow or PBL samples from patients with a variety of lymphoproliferative disorders were examined, only 2 specimens reacted with the antiserum, both from individuals with diagnoses of acute lymphocytic leukemia (ALL). None of the 14 normal bone marrow or PBL donor specimens tested reacted with the anti-AML. It was also found that essentially all samples from patients in clinical remission from AML had high numbers of cells reactive with the anti-AML. When cells from such individuals were labelled and sorted on the FACS IV, it was found that the cell population fluorescing strongly with the anti-AML contained cells of both myeloid and lymphoid origin. The AML antigen was used to produce AML specific monoclonal antibody. Spleens from AML-antigen immunized Balb/c mice were fused to NS-1 myeloma parental cells and a myelogenous leukemia specific monoclonal antibody was selected from the hybrid colonies produced. This monoclonal antibody (MAL-1) as well as the rabbit anti-AML has been used to identify myelogenous leukemia patient samples in the FACS IV. In addition, this monoclonal also demonstrates positive fluorescence binding to HL-60 (a promyelocytic leukemia cell line), while there is no binding to lymphocytic leukemia cell lines, CCRF-SB-ALL-B and CCRF-CEM-ALL-T. The MAL-1 monoclonal has been shown to be specific for myelogenous leukemia cell extracts in the ELISA and has been successfully used as an immunoadsorbent for the isolation of the AML antigen from cell extracts. No equivalent antigen was found when cell extracts from normal cells, lymphocytic leukemia cells and lymphoma cells were similarly absorbed. These findings indicate that both the rabbit anti-AML serum and MAL-1 monoclonal show specificity for an antigen associated with myelogenous leukemia cells. / Science, Faculty of / Microbiology and Immunology, Department of / Graduate

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

Leung Po-Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 204-235). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.vi / 撮要 --- p.x / TABLE OF CONTENTS --- p.xiii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis & Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis --- p.1 / Chapter 1.1.2 --- Leukemia - Aberrant Hematopoiesis --- p.4 / Chapter 1.1.2.1 --- Classification and Epidemiology of Leukemia --- p.4 / Chapter 1.1.2.2 --- Pathophysiology and Etiology of Leukemia --- p.7 / Chapter 1.1.2.3 --- Conventional Treatments for Leukemia --- p.9 / Chapter 1.1.2.4 --- New Avenues for Leukemia Therapy --- p.11 / Chapter 1.2 --- Coumarins: General Properties and Pharmacological Activities --- p.13 / Chapter 1.2.1 --- Introduction to Coumarins --- p.13 / Chapter 1.2.1.1 --- Historical Development of Coumarins --- p.13 / Chapter 1.2.1.2 --- Occurrence and Functions of Coumarins in Plants --- p.13 / Chapter 1.2.2 --- Phytochemistry and Metabolism of Coumarins --- p.14 / Chapter 1.2.2.1 --- Chemical Structures of Coumarins --- p.14 / Chapter 1.2.2.2 --- Biosynthesis of Coumarins --- p.18 / Chapter 1.2.2.3 --- Toxicology of Coumarins --- p.18 / Chapter 1.2.2.4 --- Metabolic Pathways and Pharmacokinetics of Coumarins --- p.19 / Chapter 1.2.3 --- Pharmacological Activities of Coumarins --- p.22 / Chapter 1.2.3.1 --- Anti-edema and Anti-inflammatory Activities --- p.22 / Chapter 1.2.3.2 --- Immunomodulatory Activity --- p.23 / Chapter 1.2.3.3 --- Anti-tumor Activity --- p.23 / Chapter 1.2.3.3.1 --- Mode of Entry of Coumarins into Tumor Cells --- p.23 / Chapter 1.2.3.3.2 --- Anti-carcinogenic Effect --- p.24 / Chapter 1.2.3.3.3 --- Anti-proliferative Activity --- p.25 / Chapter 1.2.3.3.4 --- Induction of Cell Differentiation --- p.26 / Chapter 1.2.3.3.5 --- Other Biological Activities --- p.26 / Chapter 1.2.4 --- Clinical Applications of Coumarins --- p.27 / Chapter 1.2.4.1 --- Treatment of Lymphoedema and Other High-protein Edemas --- p.27 / Chapter 1.2.4.2 --- Treatment of Thermal Injuries --- p.27 / Chapter 1.2.4.3 --- Therapeutic Agent for Renal Cell Carcinoma --- p.28 / Chapter 1.2.4.4 --- Therapy of Prostate Cancer --- p.29 / Chapter 1.3 --- Tumor Models Used in This Study --- p.30 / Chapter 1.3.1 --- Myeloid Leukemias --- p.30 / Chapter 1.3.1.1 --- HL-60 --- p.30 / Chapter 1.3.1.2 --- K562 --- p.30 / Chapter 1.3.1.3 --- EoL-1 --- p.30 / Chapter 1.3.1.4 --- WEHI-3B JCS --- p.31 / Chapter 1.3.2 --- Neuroblastoma - Neuro-2a BU-1 --- p.31 / Chapter 1.4 --- Aims and Scopes of This Investigation --- p.33 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.36 / Chapter 2.1.1 --- Animals --- p.36 / Chapter 2.1.2 --- Cell Lines --- p.36 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.37 / Chapter 2.1.4 --- [methyl-3H] Thymidine (3H-TdR) --- p.40 / Chapter 2.1.5 --- Methylthiazoletetrazolium (MTT) --- p.41 / Chapter 2.1.6 --- Nitro Blue Tetrazolium (NBT) --- p.41 / Chapter 2.1.7 --- Liquid Scintillation Cocktail --- p.42 / Chapter 2.1.8 --- Reagents and Buffers for Flow Cytometry --- p.42 / Chapter 2.1.9 --- Mouse Anti-MAP-2 Monoclonal Antibody --- p.44 / Chapter 2.1.10 --- Reagents for DNA Extraction --- p.44 / Chapter 2.1.11 --- Reagents for Total RNA Isolation --- p.45 / Chapter 2.1.12 --- Reagents and Buffers for RT-PCR --- p.46 / Chapter 2.1.13 --- Reagents and Buffers for Gel Electrophoresis --- p.49 / Chapter 2.1.14 --- Reagents and Buffers for Western Blot Analysis --- p.50 / Chapter 2.1.15 --- Reagents for Measuring Caspase Activity --- p.58 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Culture of the Tumor Cell Lines --- p.61 / Chapter 2.2.2 --- "Isolation, Preparation and Culture of Mouse Peritoneal Macrophages" --- p.61 / Chapter 2.2.3 --- Determination of Cell Proliferation by [3H]-TdR Incorporation Assay --- p.62 / Chapter 2.2.4 --- Determination of Cell Viability --- p.62 / Chapter 2.2.5 --- Cell Morphology Study --- p.63 / Chapter 2.2.6 --- Immunocytochemistry --- p.64 / Chapter 2.2.7 --- Confocal Microscopy --- p.64 / Chapter 2.2.8 --- NBT Reduction Assay --- p.65 / Chapter 2.2.9 --- In vivo Tumorigenicity Assay --- p.65 / Chapter 2.2.10 --- In vivo Anti-tumor Study --- p.65 / Chapter 2.2.11 --- Measurement of In vivo Macrophage Migration --- p.66 / Chapter 2.2.12 --- Measurement of Cytokine Production by ELISA --- p.66 / Chapter 2.2.13 --- Measurement of Apoptosis by DNA Fragmentation Analysis --- p.67 / Chapter 2.2.14 --- Determination of the Mitochondrial Membrane Potential --- p.67 / Chapter 2.2.15 --- Cell Cycle/DNA Content Evaluation --- p.68 / Chapter 2.2.16 --- Nitric oxide/Annexin V-PE Dual Sensor Assay --- p.68 / Chapter 2.2.17 --- Gene Expression Study --- p.68 / Chapter 2.2.18 --- Protein Expression Study --- p.71 / Chapter 2.2.19 --- Measurement of Caspase Activity --- p.74 / Chapter 2.2.20 --- Statistical Analysis --- p.75 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-TUMOR ACTIVITIES OF COUMARINS ON MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.76 / Chapter 3.2 --- Results --- p.18 / Chapter 3.2.1 --- Differential Anti-proliferative Effect of Coumarins on Various Leukemic Cell Lines In Vitro --- p.78 / Chapter 3.2.2 --- Cytotoxic Effect of Coumarins on Various Leukemic Cell Lines In Vitro --- p.91 / Chapter 3.2.3 --- "Kinetic, Reversibility and Stability Studies of the Anti- proliferative Effect of Coumarins on the Leukemia JCS cells" --- p.94 / Chapter 3.2.4 --- Induction of DNA Fragmentation in Myeloid Leukemia Cells by Coumarins --- p.100 / Chapter 3.2.5 --- Effect of Coumarins on the Cell Cycle Kinetics of the Leukemia JCS Cells In Vitro --- p.107 / Chapter 3.2.6 --- Effect of Coumarins on the In Vivo Tumorigenicity of the Leukemia JCS Cells --- p.112 / Chapter 3.2.7 --- Effect of Esculetin on the In Vivo Growth of the Leukemia JCS cells in Syngeneic Mice --- p.115 / Chapter 3.3 --- Discussion --- p.117 / Chapter CHAPTER 4: --- AN INVESTIGATION ON THE DIFFERENTIATION- INDUCING EFFECT OF COUMARINS / Chapter 4.1 --- Introduction --- p.122 / Chapter 4.2 --- Results --- p.124 / Chapter 4.2.1 --- The Differentiation-inducing Effect of Coumarins on Myeloid Leukemia Cells --- p.124 / Chapter 4.2.1.1 --- Morphological Changes in Coumarin-treated HL-60 Cells --- p.124 / Chapter 4.2.1.2 --- NBT Reduction of HL-60 Cells --- p.127 / Chapter 4.2.1.3 --- Effects of Coumarins on the Cell Size and Granularity of HL-60 Cells --- p.129 / Chapter 4.2.2 --- The Anti-proliferative and Differentiation-inducing Effects of Coumarins on Neuroblastoma Cells --- p.131 / Chapter 4.2.2.1 --- Anti-proliferative Effect of Coumarins on the BU-1 Cell Line In Vitro --- p.131 / Chapter 4.2.2.2 --- Morphological Changes in Coumarin-treated BU-1 Cells --- p.134 / Chapter 4.2.2.3 --- Immunocytochemistry of Coumarin-treated BU-1 Cells --- p.137 / Chapter 4.3 --- Discussion --- p.139 / Chapter CHAPTER 5: --- MECHANISTIC STUDIES ON THE ANTI-LEUKEMIC ACTIVITIES OF COUMARINS / Chapter 5.1 --- Introduction --- p.142 / Chapter 5.2 --- Results --- p.147 / Chapter 5.2.1 --- Modulatory Effects of Coumarins on the Expression of Apoptosis-regulatory Genes in the Leukemia JCS Cells --- p.147 / Chapter 5.2.2 --- Modulatory Effects of Coumarins on the Expression of Growth-related Genes in the Leukemia JCS Cells --- p.151 / Chapter 5.2.3 --- Modulatory Effects of Coumarins on the Expression of Apoptosis-regulatory Proteins in Leukemia JCS Cells --- p.157 / Chapter 5.2.4 --- Modulatory Effects of Coumarins on the Expression of Growth-related Proteins in Leukemia JCS Cells --- p.162 / Chapter 5.2.5 --- Effect of Coumarins on the Mitochondrial Membrane Depolarization of the Leukemia JCS cells --- p.165 / Chapter 5.2.6 --- Induction of Apoptosis and Nitric Oxide Production in Leukemia JCS Cells by Coumarins --- p.168 / Chapter 5.2.7 --- Effects of Coumarins on the Caspase Activity in the Leukemia JCS cells --- p.172 / Chapter 5.3 --- Discussion --- p.177 / Chapter CHAPTER 6: --- STUDIES ON THE IMMUNOMODULATORY EFFECT OF COUMARINS ON MURINE MACROPHAGES / Chapter 6.1 --- Introduction --- p.185 / Chapter 6.2 --- Results --- p.188 / Chapter 6.2.1 --- Effect of Coumarins on the Viability of Macrophages In vitro --- p.188 / Chapter 6.2.2 --- Effect of Coumarins on the In vivo Migration of Macrophages --- p.190 / Chapter 6.2.3 --- Effect of Coumarins on Cytokine Production by Macrophages --- p.192 / Chapter 6.3 --- Discussion --- p.194 / Chapter CHAPTER 7: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.197 / REFERENCES --- p.204

by Chan Shuk Chong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 135-142). / Statement --- p.i / Acknowledgment --- p.ii / Abbreviations --- p.iii / Abstract --- p.iv / Chapter Chapter 1: --- General Introduction / Chapter 1.1 --- Haematopoiesis --- p.1 / Chapter 1.1.1 --- Sites of haematopoiesis / Chapter 1.1.1.1 --- Bone marrow stroma / Chapter 1.1.1.2 --- Thymus / Chapter 1.1.1.3 --- Spleen and lymph node / Chapter 1.1.1.3.1 --- Spleen / Chapter 1.1.1.3.2 --- Lymph Nodes / Chapter 1.1.2 --- Blood Cell / Chapter 1.1.2.1 --- Development of T and B cells / Chapter 1.1.2.1.1 --- T cells / Chapter 1.1.2.1.2 --- B cells / Chapter 1.1.2.2 --- Development of Granulocytes and monocytes / Chapter 1.2 --- White Cell Disorder -Leukemia --- p.13 / Chapter 1.2.1 --- Leukemia - general concept / Chapter 1.2.1.1 --- Classification of leukemia / Chapter 1.2.1.2 --- Pathophysiology and Clinical features / Chapter 1.2.1.3 --- Etiology of myeloid leukemia / Chapter 1.2.2 --- Genetic basis of leukemia / Chapter 1.3 --- Acute myeloid leukemia (AML) cell model --- p.19 / Chapter 1.3.1 --- Cell Model for human acute myeloid leukemia / Chapter 1.3.2 --- Murine leukemia cell lines / Chapter 1.4 --- Induction of leukemia cell differentiation --- p.21 / Chapter 1.4.1 --- Overview of different inducers / Chapter 1.4.2 --- Cytokines as Inducers / Chapter 1.5 --- Objectives and Research Strategy --- p.26 / Chapter 1.5.1 --- Objectives / Chapter 1.5.2 --- Research strategy / Chapter Chapter 2 : --- Materials and Methods / Chapter 2.1 --- Materials --- p.29 / Chapter 2.1.1 --- Cell line / Chapter 2.1.2 --- Tissue culture medium / Chapter 2.1.3 --- Tumor necrosis Factor - alpha (TNF-α) / Chapter 2.1.4 --- Interleukin 1- alpha (IL-lα)and Interleukin 1- beta (IL-1β) / Chapter 2.1.5 --- "Monoclonal hamster anti-mouse IL-lα monoclonal hamster anti-mouse IL-1β, and Polyclonal rabbit anti-mouse TNF-α antibodies" / Chapter 2.1.6 --- Lipopolysaccharides (LPS) / Chapter 2.1.7 --- Buffers and solutions / Chapter 2.2 --- Methods : --- p.33 / Chapter 2.2.1 --- Cell culture / Chapter 2.2.2 --- Cytotoxicity assay / Chapter 2.2.3 --- Proliferation assay / Chapter 2.2.4 --- Cell morphology / Chapter 2.2.5 --- Phagocytosis assay / Chapter 2.2.6 --- Preparation of undifferentiated and differentiated murine leukemia WEHI3B (JCS) cells for cell lysate / Chapter 2.2.7 --- Isolation of total cellular RNA / Chapter 2.2.8 --- Extraction of the total RNA / Chapter 2.2.9 --- Spectrophotometry / Chapter 2.2.10 --- Electrophoresis of RNA in agarose gel containing formaldehyde / Chapter 2 2.11 --- First strand cDNA synthesis / Chapter 2.2.12 --- Cytokines phenotyping of the uninduced and induced WEHI 3B (JCS) by The Reverse Trancription Polymerase Chain Reaction method / Chapter 2.2.13 --- Gel electrophoresis of PCR- product / Chapter 2.2.14 --- Southern blot / Chapter 2.2.15 --- Dot blot / Chapter 2.2.16 --- Hybridization with oligonucleotides / Chapter 2.2.17 --- Chemiluminescent detection / Chapter Chapter 3 : --- Growth Inhibitory and Differentiation Effects of Lipopolysaccharides ( LPS ) on WEHI 3B (JCS) cells / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.1.1 --- Chemical structure of LPS / Chapter 3.1.2 --- Biological activity of LPS / Chapter 3.2 --- Results --- p.55 / Chapter 3.2.1 --- Anti-proliferative effects of LPS / Chapter 3.2.2 --- Differentiation inducing effect of LPS on WEHI 3B (JCS) cells / Chapter 3.2.3 --- Phagocytic activity LPS treated WEHI 3B (JCS) cells / Chapter 3.2.4 --- Anti-proliferative effect of TNF-α / Chapter 3.2.5 --- Differentiation inducing effect of TNF-α / Chapter 3.2.6 --- Phagocytic activity of TNF-α treated WEHI3B (JCS) cells / Chapter 3.3 --- Discussion --- p.67 / Chapter 3.4 --- Summary --- p.69 / Chapter Chapter 4 : --- The Cytokine Genes Expression of the TNF-α and LPS Treated WEHI 3B (JCS) cells / Chapter 4.1 --- Introduction --- p.70 / Chapter 4.1.1 --- Differentiation of leukemia cell line / Chapter 4.1.2 --- Study of the cytokine genes expression of WEHI 3B (JCS) cells / Chapter 4.2 --- Results --- p.72 / Chapter 4.2.1 --- Isolation of total RNA from uniduced and induced WEHI 3B (JCS) cells / Chapter 4.2.2 --- The cytokine genes expression during differentiation / Chapter 4.2.2.1 --- "Up-regulation of IL-lα, IL-1β,TNF-α and IFN-γ in both TNF-α induced and LPS induced WEHI 3B (JCS) cells" / Chapter 4.2.2.1.1 --- Southern blot / Chapter 4.2.2.1.2 --- Semi-quantitation of PCR-products by gel electrophoresis and dot-blot hybridization / Chapter 4.2.2.2 --- up-regulation of GM-CSF and G-CSF in LPS induced WEHI 3B (JCS) cells / Chapter 4.3 --- Discussion --- p.92 / Chapter 4.4 --- Summary --- p.95 / Chapter Chapter 5 : --- Growth inhibitory and Differentiation Inducing Effect of IL-l( IL-1α and IL-1β) on WEHI 3B (JCS) cells / Chapter 5.1 --- Introduction --- p.96 / Chapter 5.1.1 --- The interleukin 1 (IL-1) family / Chapter 5.1.1.1 --- Structure of IL-1 / Chapter 5.1.1.2 --- The biological function of IL-1 / Chapter 5.1.2 --- Tumor necrosis factor - alpha ( TNF-α) / Chapter 5.1.2.1 --- Structure of TNF-α / Chapter 5.1.2.2 --- Biological functions of TNF-α / Chapter 5.1.3 --- The similarity between TNF and IL-1 / Chapter 5.2 --- Results --- p.102 / Chapter 5.2.1 --- Anti-proliferative effect of IL-1 / Chapter 5.2.2 --- Differentiation inducing effect of IL-1 / Chapter 5.2.3 --- Phagocytic activity of IL-1 treated JCS cells / Chapter 5.2.4 --- "Role of endogenously produced IL-lα, IL-1β and TNF-α in LPS cytokines differentiation of WEHI 3B (JCS) cells" / Chapter 5.2.4.1. --- "Effect of neutralizing anti- ILl-α,anti - IL-l-β, and anti-TNF-α antibodies on the growth inihbition of the treated WEHI 3B (JCS) cells" / Chapter 5.2.4.2 --- "Effects of neutralizing anti-IL-lα, anti- IL-1β, and anti-TNF-α antibodies on differentiation of the treated WEHI 3B (JCS) cells" / Chapter 5.3 --- Discussion --- p.124 / Chapter 5.4 --- Summary --- p.127 / Chapter Chapter 6 --- : Concluding Discussion --- p.128 / References --- p.135

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [48]-59).

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Chan Sin-Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 154-169). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iv / Abstract (Chinese Version) --- p.vi / Contents --- p.viii / Abbreviations --- p.xiii / List of Figures and Tables --- p.xvi / Chapter Chapter One --- General Introduction / Chapter 1.1 --- Hematopoiesis --- p.1 / Chapter 1.1.1 --- Ontogeny of hematopoiesis --- p.1 / Chapter 1.1.2 --- Hiercharay of hematopoiesis --- p.2 / Chapter 1.2 --- Regulation of hematopoiesis --- p.5 / Chapter 1.2.1 --- Bone marrow stromal cell --- p.5 / Chapter 1.2.2 --- Hematopoietic growth factor --- p.6 / Chapter 1.2.3 --- Hematopoietic growth factor receptors and signal transduction --- p.10 / Chapter 1.2.4 --- Transcriptional regulation of myeloid cell development --- p.11 / Chapter 1.3 --- Deregulated hematopoiesis - Leukemia --- p.20 / Chapter 1.3.1 --- Classification of leukemia --- p.20 / Chapter 1.3.2 --- Molecular basis of leukemia --- p.20 / Chapter 1.4 --- Prostaglandin E2 induced WEHI-3B JCS and JCS-14 cell differentiation --- p.22 / Chapter 1.4.1 --- Induced leukemia cell differentiation --- p.22 / Chapter 1.4.2 --- Inducer of cell differentiation - Prostaglandin E2 --- p.22 / Chapter 1.4.3 --- WEHI-3B JCS and subline JCS-14 cells --- p.24 / Chapter 1.5 --- The aims of study --- p.26 / Chapter Chapter Two --- Identification of differentially expressed genes during PGE2-induced WEHI-3B JCS-14 cell differentiation / Chapter 2.1 --- Introduction --- p.27 / Chapter 2.1.1 --- Strategy for studying PGE2-induced JCS-14 cell differentiation --- p.28 / Chapter 2.1.2 --- Method for studying differential gene expression: Microarry Technology --- p.29 / Chapter 2.2 --- Materials --- p.32 / Chapter 2.2.1 --- Cell line --- p.32 / Chapter 2.2.2 --- AtlasT M Mouse cDNA Expression Array --- p.32 / Chapter 2.2.3 --- Chemicals --- p.32 / Chapter 2.2.4 --- Solutions and buffers --- p.33 / Chapter 2.2.5 --- Reagents --- p.34 / Chapter 2.3 --- Methods --- p.35 / Chapter 2.3.1 --- Morphological study of PGE2-induced JCS-14 cell differentiation --- p.35 / Chapter 2.3.2 --- Preparation of total RNA from PGE2-induced JCS-14 cells --- p.35 / Chapter 2.3.2.1 --- Preparation of cell lysates --- p.35 / Chapter 2.3.2.2 --- Isolation of total RNA --- p.35 / Chapter 2.3.3 --- Preparation of cDNA probes --- p.36 / Chapter 2.3.3.1 --- Probe synthesis from total RNA --- p.36 / Chapter 2.3.3.2 --- Purification of the labeled cDNA probes --- p.37 / Chapter 2.3.4 --- Hybridization cDNA probes to the Atlas Array and stringency wash --- p.37 / Chapter 2.4 --- Results --- p.39 / Chapter 2.4.1 --- Morphological changes in PGE2-treated JCS-14 cells --- p.39 / Chapter 2.4.2 --- Analysis of total RNA from PGE2-induced JCS-14 cells --- p.43 / Chapter 2.4.3 --- Hybridization of cDNA probes to AtlasT M cDNA Expression Array --- p.45 / Chapter 2.5 --- Discussion --- p.73 / Chapter 2.5.1 --- Morphological study of JCS-14 cell differentiation --- p.73 / Chapter 2.5.2 --- Differentiation commitment of JCS-14 cell under PGE2 induction --- p.73 / Chapter 2.5.3 --- Gene expression profile by microarray --- p.74 / Chapter 2.5.4 --- Gene expression profile of 5 hours PGE2-induced JCS-14 cells --- p.74 / Chapter 2.5.5 --- Further analysis of regulatory genes in PGE2-induced JCS-14 cell differentiation --- p.77 / Chapter Chapter Three --- Expression profile of identified genes in WEHI-3B JCS-14 and JCS cell differentiation / Chapter 3.1 --- Introduction --- p.79 / Chapter 3.1.1 --- Quantitation of mRNA by Real time RT-PCR --- p.80 / Chapter 3.1.2 --- Relative quantitation of gene expression --- p.83 / Chapter 3.2 --- Materials --- p.85 / Chapter 3.2.1 --- Cell lines --- p.85 / Chapter 3.2.2 --- SYBR® Green PCR core kit --- p.85 / Chapter 3.2.3 --- Chemicals --- p.85 / Chapter 3.2.4 --- Solutions and buffers --- p.86 / Chapter 3.2.5 --- Enzymes and nucleic acids --- p.86 / Chapter 3.3 --- Methods --- p.88 / Chapter 3.3.1 --- Preparation of total RNA from PGE2-induced JCS-14 and JCS cells --- p.88 / Chapter 3.3.1.1 --- Preparation of cell lysates --- p.88 / Chapter 3.3.1.2 --- Isolation of total RNA --- p.88 / Chapter 3.3.2 --- Reverse transcription (RT) --- p.88 / Chapter 3.3.3 --- Design of real-time PCR primers --- p.88 / Chapter 3.3.4 --- Determination of relative efficiency of target and reference amplification by validation experiment --- p.89 / Chapter 3.3.5 --- Confirmation of expression profile of identified genes in JCS-14 and JCS cells by comparative CT method in real-time PCR --- p.90 / Chapter 3.4 --- Results --- p.91 / Chapter 3.4.1 --- Analysis of total RNA from PGE2-induced JCS-14 and JCS cells --- p.91 / Chapter 3.4.2 --- Validation experiment of real-time PCR primers --- p.93 / Chapter 3.4.3 --- Expression profile of specific genes in JCS-14 and JCS cells by comparative CT method --- p.101 / Chapter 3.5 --- Discussion --- p.114 / Chapter 3.5.1 --- Study of gene expression profiles in JCS-14 and JCS cell differentiation --- p.114 / Chapter 3.5.2 --- Transcription analysis by real-time PCR --- p.114 / Chapter 3.5.3 --- Gene expression profiles during PGE2-induced JCS-14 and JCS cell differentiation --- p.115 / Chapter Chapter Four --- Inhibition of specific gene expression in WEHI-3B JCS-14 and JCS cells using antisense blocking technique / Chapter 4.1 --- Introduction --- p.121 / Chapter 4.1.1 --- Antisense technique --- p.122 / Chapter 4.1.2 --- Design of antisense oligonucleotides --- p.125 / Chapter 4.1.3 --- Transfer of oligonucleotides to cells --- p.128 / Chapter 4.2 --- Materials --- p.129 / Chapter 4.2.1 --- Cell lines --- p.129 / Chapter 4.2.2 --- Chemicals --- p.129 / Chapter 4.2.3 --- Reagents --- p.129 / Chapter 4.2.4 --- Solutions --- p.129 / Chapter 4.3 --- Methods --- p.131 / Chapter 4.3.1 --- Design of antisense oligonucleotides --- p.131 / Chapter 4.3.2 --- Transfection of oligonucleotides into cells --- p.134 / Chapter 4.3.3 --- Morphological study of PGE2-induced JCS-14 and JCS cells --- p.134 / Chapter 4.4 --- Results --- p.135 / Chapter 4.4.1 --- Effect of antisense oligonucleotides on JCS-14 cell differentiation --- p.135 / Chapter 4.4.2 --- Effect of antisense oligonucleotides on JCS cell differentiation --- p.136 / Chapter 4.5 --- Discussion --- p.146 / Chapter 4.5.1 --- Effects of antisense B-myb on JCS-14 and JCS cell differentiation --- p.146 / Chapter 4.5.2 --- Effects of antisense thyroid hormone receptor (c-erbA) and transcription terminator factor (TTF-1) on JCS-14 and JCS cell differentiation --- p.147 / Chapter Chapter Five --- General Discussion / Chapter 5.1 --- Introduction --- p.148 / Chapter 5.2 --- Differentiation program triggered by Prostaglandin E2 --- p.148 / Chapter 5.2.1 --- Lineage preference during differentiation --- p.148 / Chapter 5.2.2 --- Differentially expressed genes during PGE2-induced JCS-14 cell differentiation --- p.149 / Chapter 5.2.3 --- Expression patterns of the three differentially expressed genes in PGE2-induced JCS-14 and JCS cells --- p.149 / Chapter 5.2.4 --- Antisense blocking during differentiation --- p.151 / Chapter 5.3 --- Further studies --- p.152 / References --- p.154

by Lau Kwok Wing, Wilson. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 166-188). / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.v / TABLE OF CONTENTS --- p.viii / Chapter CHAPTER 1 : --- GENERAL INTRODUCTION --- p.1 / Chapter (1.1) --- Hematopoiesis : An Overview --- p.2 / Chapter (1.1.1) --- Development of Blood Cells and Sites of Hematopoiesis --- p.2 / Chapter (1.1.2) --- Hematopoietic Cytokine Network --- p.4 / Chapter (1.1.3) --- Molecular Control of Hematopoietic Cell Development --- p.5 / Chapter (1.2) --- Leukemia : An Overview --- p.8 / Chapter (1.2.1) --- Leukemia : Abnormalities in Blood Cell Formation --- p.8 / Chapter (1.2.2) --- Pathophysiology and Etiology of Leukemia --- p.10 / Chapter (1.2.3) --- New Avenues for Therapy : Induction of Differentiation and Apoptosis --- p.12 / Chapter (1.3) --- Induction of Differentiation in Myeloid Leukemia Cells --- p.14 / Chapter (1.3.1) --- Inducers of Leukemic Cell Differentiation --- p.14 / Chapter (1.3.2) --- Cytokines as Inducers of Myeloid Leukemic Cell Differentiation --- p.17 / Chapter (1.3.3) --- Phenotypic Changes and Functional Characterizations --- p.20 / Chapter (1.3.4) --- Modulation of Gene Expression in Myeloid Leukemic Cell Differentiation --- p.22 / Chapter (1.4) --- Apoptosis and Leukemic Cell Death --- p.23 / Chapter (1.4.1) --- Apoptosis : An Overview --- p.23 / Chapter (1.4.2) --- Cytokines and Apoptosis in Myeloid Leukemia --- p.26 / Chapter (1.5) --- Objectives and Research Strategy --- p.27 / Chapter (1.5.1) --- The Murine Myeloid Leukemia Cell Line (Ml) as an Experimental Cell Model for Acute Myeloid Leukemia --- p.27 / Chapter (1.5.2) --- Leukemia Inhibitory Factor (LIF) as a Differentiation Inducer --- p.28 / Chapter (1.5.3) --- Aims and Scopes of This Investigation --- p.31 / Chapter CHAPTER 2 ： --- MATERIALS AND METHODS --- p.33 / Chapter (2.1) --- Materials --- p.34 / Chapter (2.1.1) --- Mice --- p.34 / Chapter (2.1.2) --- Cell Lines --- p.34 / Chapter (2.1.3) --- Recombinant Cytokines --- p.34 / Chapter (2.1.4) --- Monoclonal Antibodies --- p.36 / Chapter (2.1.5) --- Oligonucleotide Primers and Internal Probes --- p.37 / Chapter (2.1.6) --- "Buffers, Culture Medium and Other Reagents" --- p.39 / Chapter (2.1.7) --- Reagents and Solutions for Gene Expression Study --- p.41 / Chapter (2.2) --- Methods --- p.46 / Chapter (2.2.1) --- Culture of Myeloid Leukemia Cell Lines --- p.46 / Chapter (2.2.2) --- Induction of Leukemic Cell Differentiation --- p.46 / Chapter (2.2.3) --- Determination of Cell Growth and Proliferation --- p.46 / Chapter (2.2.4) --- Cell Morphological Study --- p.47 / Chapter (2.2.5) --- Assessment of Differentiation-Associated Characteristics --- p.48 / Chapter (2.2.5.1) --- Nitroblue Tetrazolium (NBT) Reduction Assay --- p.48 / Chapter (2.2.5.2) --- Phagocytosis Assay --- p.48 / Chapter (2.2.5.3) --- Assay of Plastic Adherence --- p.49 / Chapter (2.2.6) --- Flow Cytometric Analysis --- p.49 / Chapter (2.2.6.1) --- Surface Antigen Immunophenotyping --- p.49 / Chapter (2.2.6.2) --- Assay of Endocytic Activity --- p.50 / Chapter (2.2.6.3) --- Assay of Non-specific Esterase Activity --- p.50 / Chapter (2.2.6.4) --- Cell Cycle / DNA Content Evaluation --- p.51 / Chapter (2.2.7) --- Gene Expression Analysis --- p.52 / Chapter (2.2.7.1) --- Preparation of Cell Lysate --- p.52 / Chapter (2.2.7.2) --- RNA Isolation --- p.52 / Chapter (2.2.7.3) --- Reverse Transcription --- p.53 / Chapter (2.2.7.4) --- Polymerase Chain Reaction (PGR) --- p.54 / Chapter (2.2.7.5) --- Agarose Gel Electrophoresis --- p.55 / Chapter (2.2.7.6) --- 3' End Labelling of Oligonucleotide Probes --- p.56 / Chapter (2.2.7.7) --- Dot Blot Hybridization --- p.56 / Chapter (2.2.7.8) --- Digoxigenin (DIG) Chemiluminescent Detection --- p.57 / Chapter (2.2.8) --- DNA Fragmentation Analysis --- p.58 / Chapter (2.2.9) --- Statistical Analysis --- p.59 / Chapter CHAPTER 3 ： --- "EFFECTS OF LEUKEMIA INHIBITORY FACTOR ON THE PROLIFERATION, DIFFERENTIATION, AND APOPTOSIS OF MURINE MYELOID LEUKEMIA Ml CELLS" --- p.60 / Chapter (3.1) --- Introduction --- p.61 / Chapter (3.2) --- Results --- p.63 / Chapter (3.2.1) --- Induction of Growth Arrest in rmLIF-Treated Ml Cells --- p.63 / Chapter (3.2.2) --- Induction of Monocytic Differentiation of Ml cells by rmLIF --- p.66 / Chapter (3.2.2.1) --- Morphological Changes --- p.66 / Chapter (3.2.2.2) --- Induction of Plastic Adherence --- p.70 / Chapter (3.2.2.3) --- Surface Antigen Immunophenotyping --- p.70 / Chapter (3.2.2.4) --- NBT-Reducing Activity of rmLIF-Treated Ml Cells --- p.76 / Chapter (3.2.2.5) --- Non-specific Esterase Activity of rmLIF-Treated Ml Cells --- p.77 / Chapter (3.2.2.6) --- Endocytic Activity of rmLIF-Treated Ml Cells --- p.78 / Chapter (3.2.2.7) --- Phagocytic Activity of rmLIF-Treated Ml Cells --- p.79 / Chapter (3.2.3) --- Induction of Differentiation-Associated DNA Fragmentation --- p.80 / Chapter (3.2.4) --- Production of Differentiation-Inducing Factors --- p.84 / Chapter (3.3) --- Discussion --- p.88 / Chapter CHAPTER 4 : --- CYTOKINE INTERACTIONS IN REGULATING THE PROLIFERATION AND DIFFERENTIATION OF MURINE MYELOID LEUKEMIA Ml CELLS --- p.94 / Chapter (4.1) --- Introduction --- p.95 / Chapter (4.2) --- Results --- p.97 / Chapter (4.2.1) --- Synergistic Effect of LIF and IL-6 on the Proliferation and Differentiation of Ml Cells --- p.97 / Chapter (4.2.2) --- Regulation of Proliferation and Differentiation of Ml Cells by LIF and OSM --- p.101 / Chapter (4.2.3) --- Effects of LIF and TNF-α on the Proliferation and Differentiation of Ml Cells --- p.104 / Chapter (4.2.4) --- Synergistic Effect of LIF and IL-1 on the Proliferation and Differentiation of Ml Cells --- p.107 / Chapter (4.3) --- Discussion --- p.115 / Chapter CHAPTER 5 ： --- MODULATION OF CYTOKINE AND CYTOKINE RECEPTOR GENE EXPRESSION IN LIF- INDUCED DIFFERENTIATION OF MURINE MYELOID LEUKEMIA Ml CELLS --- p.120 / Chapter (5.1) --- Introduction --- p.121 / Chapter (5.2) --- Results --- p.123 / Chapter (5.3) --- Discussion --- p.152 / Chapter CHAPTER 6 ： --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.158 / REFERENCES --- p.166

Liu, Xiaozhuo. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 156-169). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese (摘要） --- p.iv / Acknowledgments --- p.vi / List of Abbreviations --- p.vii / Table of Contents --- p.xiv / Chapter Chapter One: --- General Introduction / Chapter 1.1 --- Hematopoiesis and Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis --- p.1 / Chapter 1.1.2 --- Leukemia --- p.6 / Chapter 1.1.2.1 --- An Overview of Leukemia --- p.6 / Chapter 1.1.2.2 --- Classification and Epidemiology of Leukemia --- p.8 / Chapter 1.1.2.3 --- Conventional Approaches to Leukemia Therapy --- p.12 / Chapter 1.1.2.4 --- Novel Approaches to Leukemia Therapy --- p.15 / Chapter 1.2 --- Sophoraflavanone G: A Bioactive Compound Isolated from Kushen --- p.18 / Chapter 1.2.1 --- An Overview of Kushen: A Traditional Chinese Medicine --- p.19 / Chapter 1.2.2 --- An Overview of Lavandulyl Flavanones --- p.22 / Chapter 1.2.3 --- Historical Development and Occurrence of Sophoraflavanone G --- p.24 / Chapter 1.2.4 --- Biological Activities of Sophoraflavanone G --- p.25 / Chapter 1.2.4.1 --- Anti-microbial and Insecticidal Activities --- p.25 / Chapter 1.2.4.2 --- Anti-tumor Activities --- p.26 / Chapter 1.2.4.3 --- Pharmacodynamics of Sophoraflavanone G --- p.27 / Chapter 1.3 --- Objectives and Scopes of the Present Study --- p.30 / Chapter Chapter Two: --- Materials and Methods / Chapter 2.1 --- Materials --- p.32 / Chapter 2.1.1 --- Animals --- p.32 / Chapter 2.1.2 --- Cell lines --- p.32 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.34 / Chapter 2.1.4 --- Reagents and Buffers for Flow Cytometry --- p.37 / Chapter 2.1.5 --- Reagents for DNA Extraction --- p.39 / Chapter 2.1.6 --- Reagents for Measuring Caspase Activity --- p.40 / Chapter 2.1.7 --- "Reagents, Buffers and Materials for Western Blotting" --- p.43 / Chapter 2.2 --- Methods --- p.48 / Chapter 2.2.1 --- Extraction and Isolation of Sophoraflavanone G from Kushen --- p.48 / Chapter 2.2.2 --- Culture of Tumor Cell Lines --- p.49 / Chapter 2.2.3 --- "Isolation, Preparation and Culturing of Human Peripheral Blood Leukocytes and Murine Bone Marrow Cells" --- p.50 / Chapter 2.2.4 --- Assays for Anti-proliferation and Cytotoxicity --- p.51 / Chapter 2.2.5 --- Determination of Anti-leukemic Activity In Vivo (In Vivo Tumorigenicity Assay) --- p.52 / Chapter 2.2.6 --- Cell Cycle Analysis by Flow Cytometry --- p.53 / Chapter 2.2.7 --- Measurement of Apoptosis-induced Activities --- p.54 / Chapter 2.2.8 --- Protein Expression Study --- p.59 / Chapter 2.2.9 --- Assessment of Differentiation-associated Characteristics --- p.64 / Chapter 2.2.10 --- Statistical Analysis --- p.65 / Chapter Chapter Three: --- Studies on the Anti-proliferative Effect of Sophoraflavanone G on Human Myeloid Leukemia Cells / Chapter 3.1 --- Introduction --- p.66 / Chapter 3.2 --- Results --- p.69 / Chapter 3.2.1 --- Structure Identification of Sophoraflavanone G Isolated from Sophora flavescens --- p.69 / Chapter 3.2.2 --- Anti-proliferative Activity of Sophoraflavanone G on Various Myeloid Leukemia Cell Lines --- p.72 / Chapter 3.2.3 --- Effect of Sophoraflavanone G on the Viability of the Human Promyelocytic Leukemia HL-60 Cells --- p.80 / Chapter 3.2.4 --- Cytotoxic Effect of Sophoraflavanone G on Primary Normal Cells In Vitro --- p.83 / Chapter 3.2.5 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Sophoraflavanone G on the Human Promyelocytic Leukemia HL-60 Cells --- p.85 / Chapter 3.2.6 --- Effect of Sophoraflavanone G on the In Vivo Tumorigenicity of the HL-60 Cells --- p.88 / Chapter 3.2.7 --- Effect of Sophoraflavanone G on the Cell Cycle Profile of the HL-60 cells In Vitro --- p.90 / Chapter 3.2.8 --- Effect of Sophoraflavanone G on the Expression of Cell Cycle-regulatory Proteins in the HL-60 Cells --- p.93 / Chapter 3.2.9 --- Anti-proliferative Effect of Sophoraflavanone G on Multidrug-resistant (MDR) Leukemia Cell Line HL-60/MX2 Cells --- p.95 / Chapter 3.3 --- Discussion --- p.101 / Chapter Chapter Four: --- Studies on the Apoptosis- and Differentiation-inducing Activities of Sophoraflavanone G on Human Myeloid Leukemia Cells / Chapter 4.1 --- Introduction --- p.109 / Chapter 4.2 --- Results --- p.114 / Chapter 4.2.1 --- Induction of DNA Fragmentation in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G --- p.114 / Chapter 4.2.2 --- Induction of Phosphatidylserine Externalization in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G as Detected by Annexin V-GFP and PI Double Staining Method --- p.116 / Chapter 4.2.3 --- Effects of Sophoraflavanone G on the Caspase Activities in the Human Promyelocytic Leukemia HL-60 Cells --- p.119 / Chapter 4.2.4 --- Induction of Mitochondrial Membrane Depolarization in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G --- p.124 / Chapter 4.2.5 --- Involvement of Bcl-2 Family Members in Sophoraflavanone G-induced Apoptosis in the Human Promyelocytic Leukemia HL-60 Cells --- p.128 / Chapter 4.2.6 --- Effects of Sophoraflavanone G on the Induction of Reactive Oxygen Species in the Human Promyelocytic Leukemia HL-60 Cells --- p.131 / Chapter 4.2.7 --- Effect of Sophoraflavanone G on the Intracellular Ca2+ Level in the Human Promyelocytic Leukemia HL-60 Cells --- p.134 / Chapter 4.2.8 --- Morphological Studies on the Sophoraflavanone G-treated Human Promyelocytic Leukemia HL-60 Cells --- p.136 / Chapter 4.2.9 --- Effect of Sophoraflavanone G on the NBT Reducing Activity of the Human Promyelocytic Leukemia HL-60 Cells --- p.138 / Chapter 4.3 --- Discussion --- p.140 / Chapter Chapter Five: --- Conclusions and Future Perspectives --- p.148 / References --- p.156