Meng Jie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references. / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Table of Contents --- p.vi / Abbreviation --- p.x / List of Figures --- p.xi / List of Tables --- p.xiii / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Black Mustard and Sinigrin --- p.2 / Chapter 1.2 --- Hepatocellular Carcinoma --- p.5 / Chapter 1.2.1 --- Different Stages of HCC --- p.6 / Chapter 1.2.2 --- Risk Factors --- p.8 / Chapter 1.2.3 --- Treatments of HCC --- p.10 / Chapter 1.3 --- Biomarkers Used to Evaluate Effects of Sinigrin on HCC --- p.12 / Chapter 1.3.1 --- AST & ALT --- p.12 / Chapter 1.3.2 --- Glutathione S Transferase -p (GST-p) --- p.13 / Chapter 1.4 --- Tumor Suppressor Genes and Oncogenes --- p.14 / Chapter 1.4.1 --- "p53, the Tumor Suppressor Gene" --- p.15 / Chapter 1.4.2 --- p53-dependent pathway --- p.15 / Chapter 1.4.2.1 --- Mdm2 --- p.16 / Chapter 1.4.2.2 --- Bax and Bcl-2 --- p.17 / Chapter 1.4.2.3 --- PCNA and p21wAF1/CIP1 --- p.18 / Chapter 1.5 --- Aim of the Project --- p.19 / Chapter Chapter 2: --- Materials and Methods --- p.20 / Chapter 2.1 --- In vitro Studies --- p.21 / Chapter 2.1.1 --- Neutral Red Assay --- p.21 / Chapter 2.1.1.1 --- Chemicals and Reagents --- p.21 / Chapter 2.1.1.2 --- Liver Cells --- p.23 / Chapter 2.1.1.3 --- Neutral Red Assay --- p.24 / Chapter 2.1.2 --- Flow Cytometery --- p.24 / Chapter 2.1.2.1 --- Chemicals and Reagents --- p.25 / Chapter 2.1.2.2 --- Flow Cytometery Analysis --- p.25 / Chapter 2.1.3 --- DNA Fragmentation --- p.26 / Chapter 2.1.3.1 --- Chemicals and Reagents --- p.26 / Chapter 2.1.3.2 --- DNA Extraction --- p.28 / Chapter 2.1.3.3 --- DNA Agarose Gel Electrophoresis --- p.29 / Chapter 2.1.4 --- cDNA Microarray --- p.29 / Chapter 2.1.4.1 --- Chemicals and Reagents --- p.30 / Chapter 2.1.4.2 --- RNA Extraction --- p.33 / Chapter 2.1.4.3 --- RNA Quantity and Quality Control --- p.34 / Chapter 2.1.4.4 --- RT-PCR --- p.35 / Chapter 2.1.4.5 --- cRNA Convention and Purification --- p.36 / Chapter 2.1.4.6 --- Hybridization --- p.37 / Chapter 2.1.4.7 --- Washing and Detection --- p.37 / Chapter 2.1.4.8 --- Data Analysis --- p.38 / Chapter 2.2 --- In vivo Studies --- p.39 / Chapter 2.2.1 --- Animal Treatment --- p.39 / Chapter 2.2.1.1 --- Chemicals and Reagents --- p.39 / Chapter 2.2.1.2 --- Chemical Carcinogens --- p.40 / Chapter 2.2.1.3 --- Promotion Stage --- p.41 / Chapter 2.2.1.4 --- Progression Stage --- p.44 / Chapter 2.2.2 --- Measurement of Serum ALT and AST Activities --- p.46 / Chapter 2.2.2.1 --- Chemicals and Reagents --- p.46 / Chapter 2.2.2.2 --- Activity Assay --- p.46 / Chapter 2.2.3 --- Histological Analysis --- p.47 / Chapter 2.2.3.1 --- Chemicals and Reagents --- p.47 / Chapter 2.2.3.2 --- Preparation of Slides --- p.49 / Chapter 2.2.3.3 --- H&E Staining --- p.49 / Chapter 2.2.3.4 --- GST-p Immuno-staining --- p.50 / Chapter 2.2.4 --- Semi-Quantitative RT-PCR Analysis of mRNA Expression --- p.53 / Chapter 2.2.4.1 --- Chemicals and Reagents --- p.53 / Chapter 2.2.4.2 --- Extraction of total RNA from rat liver --- p.53 / Chapter 2.2.4.3 --- Quantity and Quality Control of RNA --- p.53 / Chapter 2.2.4.4 --- RT-PCR (Reverse Transcription) --- p.54 / Chapter 2.2.4.5 --- PCR --- p.54 / Chapter 2.2.4.6 --- DNA gel electrophoresis --- p.55 / Chapter 2.2.4.7 --- Data Analysis --- p.56 / Chapter 2.2.5 --- Western Blot Analysis for Biomarkers --- p.56 / Chapter 2.2.5.1 --- Chemicals and Reagents --- p.56 / Chapter 2.2.5.2 --- Extraction of the Cytosol Protein --- p.60 / Chapter 2.2.5.3 --- Extraction of the Nuclear protein --- p.61 / Chapter 2.2.5.4 --- SDS Gel Electrophoresis --- p.61 / Chapter 2.2.5.5 --- Western Blot --- p.62 / Chapter 2.2.5.6 --- Interaction with Antibodies --- p.63 / Chapter 2.2.5.7 --- ECL Detection --- p.63 / Chapter 2.2.5.8 --- Data Analysis --- p.64 / Chapter Chapter 3: --- Results --- p.65 / Chapter 3.1 --- In vitro Studies --- p.66 / Chapter 3.1.1 --- Cell Viability test and IC50 --- p.66 / Chapter 3.1.2 --- Cell Cycle Analysis --- p.68 / Chapter 3.1.3 --- DNA Fragmentation --- p.71 / Chapter 3.1.4 --- Effects of Sinigrin on Gene Expression --- p.73 / Chapter 3.2 --- In vivo Studies --- p.77 / Chapter 3.2.1 --- Effects of Sinigrin on HCC Development (Promotion stage) in Rats --- p.77 / Chapter 3.2.1.1 --- Direct Observation --- p.77 / Chapter 3.2.1.2 --- Relative Liver / Body Weight Ratio --- p.79 / Chapter 3.2.1.3 --- AST/ALT Assay --- p.81 / Chapter 3.2.1.4 --- Basic Structure of Hepatocytes --- p.83 / Chapter 3.2.1.5 --- GST-p Foci Area --- p.85 / Chapter 3.2.1.6 --- mRNA Expression of p53 and Mdm2 --- p.88 / Chapter 3.2.1.7 --- Protein Expression of Biomarkers --- p.90 / Chapter 3.2.2 --- Effects of Sinigrin on HCC Development (Progression stage) in Rats --- p.97 / Chapter 3.2.2.1 --- Direct Observation --- p.97 / Chapter 3.2.2.2 --- Relative Liver / Body Weight Ratio --- p.99 / Chapter 3.2.2.3 --- AST/ALT Assay --- p.101 / Chapter 3.2.2.4 --- Basic Structure of Hepatocytes --- p.103 / Chapter 3.2.2.5 --- GST-p Foci Area --- p.105 / Chapter 3.2.2.6 --- mRNA Expression of p53 and Mdm2 --- p.108 / Chapter 3.2.2.7 --- Protein Expression of Biomarkers --- p.110 / Chapter Chapter 4: --- Discussion --- p.116 / Chapter 4.1 --- Protective and Therapeutic Benefits of Sinigrin --- p.117 / Chapter 4.1.1 --- Effects of SIN on Cancer and Normal Cells --- p.117 / Chapter 4.1.2 --- Effective Tumor Induction by DEN-CC14 Treatment --- p.118 / Chapter 4.1.3 --- Protective Effect of SIN in the Promotion Stage of HCC --- p.118 / Chapter 4.1.4 --- Therapeutic Effect of SIN in the Progression Stage of HCC --- p.119 / Chapter 4.2 --- Biological Activities of SIN --- p.121 / Chapter 4.3 --- Summary --- p.134 / References --- p.xiv

Yip, Wai Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 203-228). / Abstracts in English and Chinese. / ACKNOWLEDGMENTS --- p.i / ABBREVIATIONS --- p.iii / ABSTRACT --- p.x / 撮要 --- p.xiv / TABLE OF CONTENTS --- p.xvii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis and Leukemia / Chapter 1.1.1 --- An Overview on Hematopoiesis Development --- p.1 / Chapter 1.1.1.1 --- Hematopoietic Growth Factors --- p.4 / Chapter 1.1.1.2 --- Site Switching of Hematopoiesis --- p.5 / Chapter 1.1.2 --- An Overview on Leukemia --- p.7 / Chapter 1.1.2.1 --- Classification of Leukemia --- p.7 / Chapter 1.1.2.2 --- Conventional Therapy of Leukemia --- p.10 / Chapter 1.1.2.3 --- Novel Approaches to Leukemia Therapy: Apoptosis and Differentiation Induction --- p.13 / Chapter 1.2 --- Polysaturated Fatty Acids / Chapter 1.2.1 --- An Overview on Polyunsaturated Fatty Acids --- p.16 / Chapter 1.2.2 --- An Overview on Essential Fatty Acids --- p.17 / Chapter 1.2.2.1 --- Alpha Linolenic Acids (ALA) --- p.17 / Chapter 1.2.2.2 --- Gamma Linolenic Acid (GLA) --- p.18 / Chapter 1.2.3 --- "An Overview on Conjugated Fatty Acids: Conjugated Linoleic Acid (CLA), Conjugated EPA and Conjugated DHA" --- p.20 / Chapter 1.2.4 --- Conjugated Linolenic Acid (CLN) --- p.24 / Chapter 1.2.4.1 --- Identification and Production of CLN --- p.28 / Chapter 1.2.4.2. --- Metabolism of CLN --- p.29 / Chapter 1.2.4.3 --- Anti-Obese and Hypolipidemic Effect of CLN --- p.30 / Chapter 1.2.4.4 --- Anti-Proliferative Effect of CLN --- p.30 / Chapter 1.2.4.5 --- Other Novel Effects of CLN --- p.32 / Chapter 1.3 --- Aims and Scopes of This Investigation --- p.34 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.36 / Chapter 2.1.1 --- Animals --- p.36 / Chapter 2.1.2 --- Human Cell Lines --- p.36 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.38 / Chapter 2.1.4 --- Reagents and Buffer for Flow Cytometry --- p.44 / Chapter 2.1.5 --- Reagents for DNA Extraction --- p.47 / Chapter 2.1.6 --- Cell Death Detection ELISApLus --- p.48 / Chapter 2.1.7 --- Reagents for Measuring Caspase Activity --- p.50 / Chapter 2.1.8 --- Reagents for FACE´ёØ ELISA Kit --- p.53 / Chapter 2.1.9 --- Reagents for Western Blotting --- p.55 / Chapter 2.2 --- Methods --- p.65 / Chapter 2.2.1 --- Culturing the Tumor Cell Lines --- p.65 / Chapter 2.2.2 --- "Isolation, Preparation and Culturing of Murine Peritoneal Macrophages and Bone Marrow Cells" --- p.66 / Chapter 2.2.3 --- Anti-proliferation Assays --- p.67 / Chapter 2.2.4 --- Cell Viability Determination --- p.68 / Chapter 2.2.5 --- Determination of Anti-leukemia Activity In Vivo (In Vivo Tumorigenicity Assay) --- p.69 / Chapter 2.2.6 --- Cell Cycle Analysis by Flow Cytometry --- p.69 / Chapter 2.2.7 --- Detection of Apoptosis --- p.70 / Chapter 2.2.8 --- Assessment of Differentiation-associated Characteristics --- p.74 / Chapter 2.2.9 --- Measurement of Caspase Activities --- p.76 / Chapter 2.2.10 --- Protein Expression Study --- p.78 / Chapter 2.2.11 --- Detection of Phosphorylation of JNK by FACE´ёØ JNK ELISA Kit --- p.83 / Chapter 2.2.12 --- Detection of Phosphorylation of NF-kB by FACE´ёØ NF-kB p65 Profiler --- p.85 / Chapter 2.2.13 --- Statistical Analysis --- p.85 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI PROLIFERATIVE EFFECTS OF CONJUGATED LINOLENIC ACIDS ON THE HUMAN MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.86 / Chapter 3.2 --- Results / Chapter 3.2.1 --- Anti-proliferative Activity of CLN Isomers on Various Myeloid Leukemia and Lymphoma Cell Lines In Vitro --- p.88 / Chapter 3.2.2 --- Direct Cytotoxic Effect of Jacaric Acid on HL-60 Cells In Vitro --- p.95 / Chapter 3.2.3 --- Cytotoxic Effect of Jacaric Acid on Primary Murine Cells and Human Normal Cell Lines In Vitro --- p.98 / Chapter 3.2.4 --- Kinetics and Reversibility Studies of the Anti-proliferative Effect of Four CLN Isomers on the Human Promyelocytic Leukemia HL-60 Cells --- p.101 / Chapter 3.2.5 --- Synergistic Anti-proliferative Effect of Jacaric Acid with Vitamin D3 and All Trans-Retinoic Acid (ATRA) on the Human Promyelocytic Leukemia HL-60 Cells In Vitro --- p.114 / Chapter 3.2.6 --- Effect of Jacaric Acid on the Cell Cycle Profile of the HL-60 Cells In Vitro --- p.116 / Chapter 3.2.7 --- Effect of Jacaric Acid on the In Vivo Tumorigenicity of the HL-60 Cells --- p.119 / Chapter 3.3 --- Discussion --- p.121 / Chapter CHAPTER 4: --- STUDIES ON THE APOPTOSIS-INDUCING AND DIFFERENTIATION-INDUCING EFFECTS OF CONJUGATED LINOLENIC ACIDS ON THE HUMAN MYELOID LEUKEMIA CELLS / Chapter 4.1.1 --- Introduction --- p.128 / Chapter 4.2 --- Results / Chapter 4.2.1 --- Induction of Apoptosis in the Human Promyelocytic Leukemia HL-60 Cells by Jacaric Acid --- p.134 / Chapter 4.2.2 --- Apoptosis-Inducing Effect of Jacaric Acid on the Human Promyelocytic Leukemia HL-60 Cells as Detected by Annexin V-GFP PI Double Staining Method --- p.138 / Chapter 4.2.3 --- Effect of Jacaric Acid on the Mitochondrial Membrane Potential in the Human Promyelocytic Leukemia HL-60 Cells --- p.140 / Chapter 4.2.4 --- Effects of Jacaric Acid on the Caspase Activities in the Human Promyelocytic Leukemia HL-60 Cells --- p.142 / Chapter 4.2.5 --- Effects of Jacaric Acid and Antioxidants on the ROS Induction in the Human Promyelocyic Leukemia hl-6 Cells --- p.147 / Chapter 4.2.6 --- Effect of N-acetyl-L-Cysteine on the Apoptosis-Inducing Activity of Jacaric Acid in the Human Promyelocytic Leukemia HL-60 Cells --- p.149 / Chapter 4.2.7 --- Morphological Studies on the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.151 / Chapter 4.2.8 --- Cell Size and Granularity of the Human Promyelocytic Leukemia HL-60 Cells after Treatment with Different CLN Isomers --- p.153 / Chapter 4.2.9 --- Expression of Differentiation-Related Cell Surface Markers in the Human Promyelocytic Leukemia HL-60 Cells after Treatment with Jacaric Acid --- p.155 / Chapter 4.3 --- Discussion --- p.158 / Chapter CHAPTER 5: --- STUDIES ON THE APOPTOSIS-ASSOCIATED PROTEINS AND SIGNALING PATHWAYS IN CONJUGATED LINOLENIC ACID-INDUCED APOPTOSIS OF THE HUMAN MYELOID LEUKEMIA CELLS / Chapter 5.1 --- Introduction --- p.165 / Chapter 5.2 --- Results / Chapter 5.2.1 --- Expression of Fas and Fas Ligand Proteins in the Jacaric Acid- treated Human Promyelocytic Leukemia HL-60 Cells --- p.171 / Chapter 5.2.2 --- Expression of Bcl-2 Family Member Proteins in the Jacaric Acid- treated Human Promyelocytic Leukemia HL-60 Cells --- p.173 / Chapter 5.2.3 --- Cytochrome c Release in the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.175 / Chapter 5.2.4 --- Cleavage of Poly(ADP-ribose) Polymerase (PARP) in the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.177 / Chapter 5.2.5 --- Phosphorylation of ERK in the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.179 / Chapter 5.2.6 --- Phosphorylation of JNK in the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.181 / Chapter 5.2.7 --- Phosphorylation of NF-kB Protein in the Jacaric Acid-treated Human Promyelocytic Leukemia HL-60 Cells --- p.183 / Chapter 5.3 --- Discussion --- p.185 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.195 / REFERENCES --- p.203

Lau Ven Gie Vengie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 113-121). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / 論文摘要 --- p.iv / TABLE OF CONTENTS --- p.vi / LIST OF FIGURES --- p.ix / LIST OF TABLES --- p.x / LIST OF ABBREVIATIONS --- p.xi / INTRODUCTION --- p.1 / Chapter I --- Hepatocellular Carcinoma --- p.1 / Risk factors --- p.1 / Stages in chemical carcinogenesis --- p.2 / Initiation --- p.2 / Promotion --- p.3 / Progression --- p.5 / Treatment of hepatocarcinoma --- p.6 / Chemotherapy ´ؤ hepatic arterial infusion (HAI) --- p.6 / Trans-arterial chemoembolization (TACE) --- p.7 / Radiofrequency ablation (RFA) --- p.8 / Percutaneous ethanol injection (PEI) --- p.9 / Liver resection --- p.9 / Liver transplantation --- p.10 / Chapter II --- Molecular Mechanisms: Oncogenes and Tumor-suppressor genes --- p.11 / Cell cycle control --- p.12 / p53 mutation in HCC --- p.13 / Normal functions of p53 and its target genes --- p.13 / p21(Wafl/Cipl/Sdil) --- p.13 / PCNA --- p.14 / Bcl-2 and Bax: the Bcl-2 family --- p.14 / Mdm2 --- p.17 / Chapter III --- Evaluation of the effects of hepatocarcinogenesis --- p.19 / GST-Pi --- p.19 / AST & ALT --- p.19 / Chapter IV --- Traditional Chinese Medicine (TCM) --- p.21 / Andrographis Paniculata --- p.21 / Pharmacological properties of andrographolide --- p.23 / Chapter V --- Aim of the project --- p.26 / MATERIALS AND METHODS --- p.27 / Chapter 1 --- Effects of andrographolide on cell viability and cell cycle --- p.27 / Chapter 1.1 --- Materials and solutions --- p.27 / Chapter 1.2 --- Preparation of solutions --- p.28 / Chapter 1.3 --- Procedures --- p.29 / Chapter 1.3.1 --- Seeding cells into culture flask --- p.29 / Chapter 1.3.2 --- Subculturing technique --- p.30 / Chapter 1.3.3 --- Neutral red assay --- p.30 / Chapter 1.3.4 --- DNA purification of HepG2 cells --- p.31 / Chapter 1.3.5 --- DNA gel electrophoresis --- p.32 / Chapter 1.3.6 --- Flow cytometry --- p.32 / Chapter 2 --- Effects of andrographolide on gene expressions --- p.33 / Chapter 2.1 --- Materials and solutions --- p.33 / Chapter 2.2 --- Preparation of solutions --- p.34 / Chapter 2.3 --- Procedures --- p.35 / Chapter 2.3.1 --- Cell treatments --- p.35 / Chapter 2.3.2 --- mRNA extraction from cell --- p.35 / Chapter 2.3.3 --- Determination of total RNA yield and quality yield --- p.36 / Chapter 2.3.4 --- RNA formaldehyde agarose gel electrophoresis --- p.36 / Chapter 2.3.5 --- cDNA synthesis --- p.37 / Chapter 2.3.6 --- "cRNA synthesis, labeling and amplification" --- p.39 / Chapter 2.3.7 --- cRNA purification --- p.40 / Chapter 2.3.8 --- Oligo GEArray hybridization --- p.41 / Chapter 2.3.9 --- Chemiluminescent detection --- p.43 / Chapter 2.3.10 --- Data analysis --- p.44 / Chapter 3 --- Effects of andrographolide on hepatocarcinogenesis in rats --- p.45 / Chapter 3.1 --- Materials and solutions --- p.45 / Chapter 3.2 --- Preparation of solutions --- p.46 / Chapter 3.3 --- Procedures --- p.47 / Chapter 3.3.1 --- Animal treatment --- p.47 / Chapter 3.3.2 --- Promotion (Experiment 1) --- p.48 / Chapter 3.3.3 --- Progression (Experiment 2) --- p.49 / Chapter 3.3.4 --- Extraction of blood serum --- p.52 / Chapter 3.3.5 --- Measurement of absorbance --- p.52 / Chapter 3.3.6 --- Tissue processing --- p.53 / Chapter 3.3.7 --- Hematoxylin and Eosin (H&E) Staining --- p.53 / Chapter 3.3.8 --- Immunohistochemical staining of GST-P --- p.54 / Chapter 3.3.9 --- Examination of liver sections --- p.55 / Chapter 4 --- "Effects of andrographolide on the expressions of Mdm2, p53, PCNA, Bax, Bcl-2 & p21" --- p.56 / Chapter 4.1 --- Materials and solutions --- p.56 / Chapter 4.2 --- Preparation of solutions --- p.57 / Chapter 4.3 --- Procedures --- p.59 / Chapter 4.3.1 --- Total mRNA extraction from liver --- p.59 / Chapter 4.3.2 --- Reverse transcription of mRNA to cDNA --- p.59 / Chapter 4.3.3 --- Protocol for polymerase chain reaction (PCR) --- p.60 / Chapter 4.3.4 --- DNA gel electrophoresis --- p.61 / Chapter 4.3.5 --- Nuclear protein extraction --- p.61 / Chapter 4.3.6 --- Cytosolic protein extraction --- p.62 / Chapter 4.3.7 --- Determination of protein concentration --- p.62 / Chapter 4.3.8 --- Immunoprecipitation of p53 from liver nuclear protein --- p.62 / Chapter 4.3.9 --- Protein gel electrophoresis by SDS-PAGE --- p.63 / Chapter 4.3.10 --- Western blotting --- p.64 / RESULTS --- p.66 / Chapter 1 --- Effects of andrographolide on cell viability and cell cycle --- p.66 / Chapter 2 --- Effects of andrographolide on gene expressions --- p.76 / Chapter 3 --- Effects of andrographolide on hepatocarcinogenesis in rats --- p.79 / Chapter 4 --- "Effects of andrographolide on the expressions of Mdm2, p53, PCNA, Bax, Bcl-2 & p21" --- p.91 / DISCUSSION --- p.102 / CONCLUSION --- p.111 / REFERENCES --- p.113

Yeung, Tak Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 181-197). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iv / Acknowledgements --- p.vi / List of Publications --- p.vii / Presentation --- p.vii / List of Abbreviations --- p.viii / Abbreviations in Figures --- p.xiii / Abbreviations in Symbols --- p.xiv / List of Cell Lines Used in this Study --- p.xv / Table of Contents --- p.xvi / List of Figures --- p.xxv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Cellular Redox State and Cancer Biology --- p.1 / Chapter 1.2 --- Reactive Oxygen Species (ROS) --- p.1 / Chapter 1.3 --- Regulation of Cellular Redox State by Intrinsic and Extrinsic Antioxidant Systems --- p.5 / Chapter 1.3.1 --- Intrinsic Antioxidant System --- p.6 / Chapter 1.3.2 --- Extrinsic Antioxidant System --- p.8 / Chapter 1.4 --- Glutathione --- p.9 / Chapter 1.4.1 --- General Information of Glutathione --- p.9 / Chapter 1.4.2 --- Functions of Glutathione --- p.12 / Chapter 1.4.2.1 --- As an Antioxidant and Free Radical Scavenger --- p.12 / Chapter 1.4.2.2 --- As a Detoxifier --- p.13 / Chapter 1.4.2.3 --- As a Regulator of Cell Signaling --- p.14 / Chapter 1.4.3 --- Synthesis of Glutathione --- p.15 / Chapter 1.4.4 --- Catabolism of Glutathione --- p.15 / Chapter 1.4.5 --- Transport and Uptake of Glutathione --- p.16 / Chapter 1.4.6 --- Glutathione in Cancer Biology --- p.18 / Chapter 1.4.6.1 --- "Role of Glutathione in the Regulation of Carcinogenesis, Growth and Apoptosis of Cancer Cells" --- p.18 / Chapter 1.4.6.1.1 --- Role of Glutathione in Carcinogenesis --- p.18 / Chapter 1.4.6.1.2 --- Role of Glutathione in the Growth of Cancer Cells --- p.20 / Chapter 1.4.6.1.3 --- Role of Glutathione in Apoptosis of Cancer Cells --- p.21 / Chapter 1.4.6.2 --- Role of Glutathione in the Regulation of Metastasis --- p.23 / Chapter 1.4.6.3 --- Role of Glutathione in Cancer Resistance and Therapy --- p.24 / Chapter 1.4.6.3.1 --- Role of Glutathione in Cancer Resistance --- p.24 / Chapter 1.4.6.3.2 --- Role of Glutathione in Cancer Therapy --- p.24 / Chapter 1.5 --- Aims of the Present Study --- p.25 / Chapter Chapter 2 --- In Vitro Study of Bαicαlein and Baicalin on Glutathione Depletion --- p.28 / Chapter 2.1 --- Introduction --- p.28 / Chapter 2.1.1 --- Scutellaria bαicαlensis Georgi --- p.28 / Chapter 2.1.1.1 --- General Clinical Applications to Treat or Prevent Diseases --- p.28 / Chapter 2.1.1.2 --- As an Antioxidant and Free Radical Scavenger --- p.29 / Chapter 2.1.1.3 --- Long History for Treatment of Cancers with the Obscure Mechanism --- p.30 / Chapter 2.1.1.4 --- Major Components --- p.31 / Chapter 2.1.2 --- Baicalein and Baicalin --- p.32 / Chapter 2.1.2.1 --- General Clinical Applications to Treat or Prevent Diseases --- p.32 / Chapter 2.1.2.2 --- As an Antioxidant and Free Radical Scavenger --- p.33 / Chapter 2.1.3 --- Hypothesis: Baicalein and Baicalin Induce Cancer Cell Death Via Glutathione Depletion --- p.35 / Chapter 2.2 --- Materials and Methods --- p.36 / Chapter 2.2.1 --- Chemicals --- p.36 / Chapter 2.2.2 --- Buffers and Solutions --- p.36 / Chapter 2.2.3 --- Animals --- p.37 / Chapter 2.2.4 --- Preparation of Rat Brain Microsomes --- p.37 / Chapter 2.2.5 --- Glutathione Depletion Assay In Vitro and Thiol Depletion Assay in Rat Brain Microsomes --- p.38 / Chapter 2.2.6 --- Statistical Analysis --- p.39 / Chapter 2.3 --- Results --- p.40 / Chapter 2.3.1 --- Effects of Baicalein and Baicalin on Sulfhydryl Contents of Glutathione --- p.42 / Chapter 2.3.2 --- Effects of Baicalein and Baicalin on Sulfhydryl Contents of Rat Brain Microsomes --- p.42 / Chapter 2.4 --- Discussion --- p.44 / Chapter Chapter 3 --- Effects of Baicalein and Baicalin on Proliferation of Different Human Cancer and Normal Cells --- p.45 / Chapter 3.1 --- Introduction-Importance of Developing A Novel Compound Inducing Cancer Cells to Cell Death with the Least Side Effects on Normal Cells --- p.45 / Chapter 3.2 --- Materials and Methods --- p.46 / Chapter 3.2.1 --- Instruments --- p.46 / Chapter 3.2.2 --- Chemicals and Cell Culture Reagents --- p.46 / Chapter 3.2.3 --- Buffers --- p.46 / Chapter 3.2.4 --- Cell Lines --- p.47 / Chapter 3.2.5 --- Cell Culture --- p.48 / Chapter 3.2.6 --- Determination of Cell Proliferation by MTT Assay --- p.49 / Chapter 3.3 --- Results --- p.51 / Chapter 3.3.1 --- Anti-Proliferative Effects of Baicalein and Baicalin on Different Cancer Cell Lines --- p.51 / Chapter 3.3.2 --- Effects of Baicalein on Different Normal Cell Lines --- p.56 / Chapter 3.4 --- Discussion --- p.58 / Chapter 3.4.1 --- Anti-Proliferative Effects of Baicalein and Baicalin on Different Cancer Cell Lines --- p.58 / Chapter 3.4.2 --- Effects of Baicalein on Cell Proliferation on Different Human Normal Cell Lines --- p.60 / Chapter Chapter 4 --- Glutathione-Depleting Effects of Baicalein on Cell Proliferation of Different Cell Lines --- p.61 / Chapter 4.1 --- Introduction-Brain Tumors --- p.61 / Chapter 4.1.1 --- Types and Classifications of Brain Tumors --- p.61 / Chapter 4.1.2 --- "Incidence Time, Patient Survival Time and Rate for" --- p.65 / Chapter 4.1.3 --- Symptoms and Diagnostic Methods for Brain Tumors --- p.66 / Chapter 4.1.4 --- "Treatments, Side Effects and Difficulties of Treatments for Brain Tumors" --- p.67 / Chapter 4.1.5 --- Glutathione Levels in Brain Normal and Cancer Cells --- p.69 / Chapter 4.2 --- Materials and Methods --- p.70 / Chapter 4.2.1 --- Instruments --- p.70 / Chapter 4.2.2 --- Chemicals --- p.70 / Chapter 4.2.3 --- Buffers --- p.70 / Chapter 4.2.4 --- Determination of Cell Proliferation by MTT Assay --- p.70 / Chapter 4.2.5 --- Determination of Intracellular Glutathione Depletion by Fluorescent Dye CMAC --- p.71 / Chapter 4.2.6 --- Determination of Cellular Reduced Glutathione Levels by DTNB-Coupled Glutathione Reductase Recycling Assay --- p.73 / Chapter 4.3 --- Results --- p.75 / Chapter 4.3.1 --- Effects of Baicalein on Intracellular GSH Levels and Cell Proliferation for Different Cell Lines --- p.75 / Chapter 4.3.2 --- Basal Intracellular Glutathione in Different Cell Lines --- p.81 / Chapter 4.4 --- Discussion --- p.84 / Chapter 4.4.1 --- Intracellular Glutathione Depletion and Cell Death Induction Effects of Baicalein on Different Cell Lines --- p.84 / Chapter 4.4.2 --- Relationship between Basal Glutathione Levels and Drug Susceptibilities --- p.85 / Chapter Chapter 5 --- Effects of Baicalein on Apoptosis and Caspase Pathways --- p.88 / Chapter 5.1 --- Introduction-Modes of Cell Death --- p.88 / Chapter 5.1.1 --- Necrosis --- p.88 / Chapter 5.1.2 --- Apoptosis --- p.89 / Chapter 5.2 --- Materials and Methods --- p.92 / Chapter 5.2.1 --- Chemicals --- p.92 / Chapter 5.2.2 --- Buffers --- p.92 / Chapter 5.2.3 --- Determination of Change of Mitochondrial Membrane Potential by JC-1 --- p.93 / Chapter 5.2.4 --- Determination of Apoptosis by Annexin V-Propidium Iodide Staining --- p.94 / Chapter 5.2.5 --- Determination of Cell Cycle Arrest by Propidium Iodide Staining --- p.95 / Chapter 5.2.6 --- "Determination of Caspase-3, -8 and -9 Activities by Fluorescent-Labeled Peptides" --- p.96 / Chapter 5.2.7 --- Determination of DNA Fragmentation --- p.97 / Chapter 5.2.8 --- Terminal Deoxynucleotidyl Transferase Mediated dUTP End Labeling (TUNEL) Assay --- p.99 / Chapter 5.2.9 --- Flow Cytometry --- p.101 / Chapter 5.3 --- Results --- p.102 / Chapter 5.3.1 --- Effects of Baicalein on Mitochondrial Membrane Potential by JC-1 Staining --- p.102 / Chapter 5.3.2 --- Effects of Baicalein on Apoptosis and Necrosis by Annexin V-Propidium Iodide Staining --- p.104 / Chapter 5.3.3 --- Effects of Baicalein on Cell Cycle Arrest by Propidium Iodide Staining --- p.108 / Chapter 5.3.4 --- "Effects of Baicalein on Caspase-3, -8 and -9 Activities" --- p.110 / Chapter 5.3.5 --- Effeets of Baiealein on DNA Fragmentation --- p.115 / Chapter 5.3.6 --- Effects of Baicalein on TUNEL Assay --- p.117 / Chapter 5.4 --- Discussion --- p.120 / Chapter Chapter 6 --- Pro-Oxidant Role of Baicalein on Reactive Oxygen Species Generation --- p.122 / Chapter 6.1 --- Introduction --- p.122 / Chapter 6.2 --- Materials and Methods --- p.122 / Chapter 6.2.1 --- Chemicals --- p.122 / Chapter 6.2.2 --- Determination of Cellular Reactive Oxygen Species Generation by Fluorescent Dye cDCFDA --- p.123 / Chapter 6.2.3 --- Determination of Mitochondrial Reactive Oxygen Species Generation by Fluorescent Dye Rhl23 --- p.124 / Chapter 6.3 --- Results --- p.125 / Chapter 6.3.1 --- Effects of Baicalein on Cellular ROS Generation by Fluorescent Dye cDCFDA --- p.125 / Chapter 6.3.2 --- Effects of Baicalein on Mitochondrial ROS Generation by Fluorescent Dye Rhl23 --- p.129 / Chapter 6.4 --- Discussion --- p.132 / Chapter Chapter 7 --- The Anticancer Mechanistic Study of Baicalein --- p.133 / Chapter 7.1 --- Introduction --- p.133 / Chapter 7.2 --- Materials and Methods --- p.134 / Chapter 7.2.1 --- Chemicals --- p.134 / Chapter 7.2.2 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death by Different Antioxidant Treatments --- p.134 / Chapter 7.2.3 --- Reversibility of Baicalein-Induced Cellular ROS Generation --- p.136 / Chapter 7.2.4 --- Reversibility of Baicalein-Induced Apoptosis by Co-Treatment of Different Antioxidants and Caspase Inhibitors --- p.137 / Chapter 7.2.5 --- "Reversibility of Baicalein-Induced Caspase-3, -8 and -9 Activation by Co-Treatment of Different Antioxidants" --- p.138 / Chapter 7.3 --- Results --- p.139 / Chapter 7.3.1 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death by Different Antioxidant Treatments --- p.139 / Chapter 7.3.1.1 --- Pre-treatments --- p.139 / Chapter 7.3.1.2 --- Co-treatments --- p.141 / Chapter 7.3.1.3 --- Post-treatments --- p.144 / Chapter 7.3.2 --- Reversibility of Baicalein-Induced Cellular ROS Generation by Co-Treatment of Different Antioxidants --- p.147 / Chapter 7.3.3 --- Reversibility of Baicalein-Induced Apoptosis by Co-Treatment of Different Antioxidants and Caspase Inhibitors --- p.152 / Chapter 7.3.4 --- Reversibility of Baicalein-Induced Caspase-3 Activation by Co-Treatment of Different Antioxidants --- p.156 / Chapter 7.3.5 --- Reversibility of Baicalein-Induced Caspase-8 and -9 Activation by Co-Treatment of Different Antioxidants --- p.160 / Chapter 7.4 --- Discussion --- p.164 / Chapter 7.4.1 --- Reversibility of Baicalein-Induced GSH Depletion and Cell Death --- p.164 / Chapter 7.4.2 --- "Reversibility of Baicalein-Induced ROS Generation," --- p.167 / Chapter 7.5 --- Concluding Remarks --- p.168 / Chapter Chapter 8 --- General Discussion --- p.169 / Chapter 8.1 --- Drug Delivery to Brain --- p.169 / Chapter 8.2 --- Protective Roles of Baicalein on Brain Cells --- p.170 / Chapter 8.2.1 --- Actions Against Oxidative Stress --- p.170 / Chapter 8.2.2 --- Actions Against Other Neurotoxic Damages --- p.171 / Chapter 8.2.3 --- Actions Against Neuronal Diseases --- p.172 / Chapter 8.3 --- Anticancer Roles of Baicalein on Astrocytoma --- p.173 / Chapter 8.4 --- Implications on the Dual Roles of Baicalein: Antioxidant and Pro-oxidant --- p.175 / Chapter 8.5 --- Future Perspectives --- p.175 / Chapter 8.5.1 --- Effects of Baicalein on Antioxidant System --- p.175 / Chapter 8.5.2 --- Effects of Baicalein on GSH Synthesis --- p.176 / Chapter 8.5.3 --- In Vivo Studies on Cytotoxic Effects of Baicalein --- p.177 / Chapter 8.5.4 --- In Vivo Studies on Anti-Tumor Effects and In Vitro Studies on Anti-Metastasis Effects of Baicalein --- p.178 / Reference List --- p.181

Zhou, Linli. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 108-118). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Abbreviations --- p.v / List of Figures --- p.vii / List of Tables --- p.ix / Table of Contents --- p.x / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Breast Cancer --- p.1 / Chapter 1.1.1 --- Introduction to Breast Cancer --- p.1 / Chapter 1.1.2 --- Types of Breast Cancer --- p.3 / Chapter 1.1.3 --- Epidemiologic Risk Factors and Etiology --- p.4 / Chapter 1.2 --- Estrogen and Breast Cancer --- p.7 / Chapter 1.3 --- Estrogen Receptor --- p.9 / Chapter 1.4 --- Current Treatment of Breast Cancer --- p.10 / Chapter 1.4.1 --- Chemotherapy --- p.10 / Chapter 1.4.2 --- Hormonal (Anti-Estrogen) Therapy --- p.11 / Chapter 1.4.2.1 --- Tamoxifen and Other Anti-estrogens --- p.12 / Chapter 1.4.2.2 --- Disadvantages of Tamoxifen --- p.13 / Chapter 1.5 --- Arsenic Trioxide --- p.14 / Chapter 1.5.1 --- The Characteristics of Arsenic Trioxide (AS2O3) --- p.14 / Chapter 1.5.2 --- The Medical use of Arsenic Trioxide (As2O3) --- p.16 / Chapter 1.5.3 --- Arsenic Trioxide (As2O3) in treating Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.1 --- Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.2 --- All-trans Retinoic Acid (ATRA) Treatment of APL --- p.18 / Chapter 1.5.3.3 --- Clinical Trial of the Arsenic Trioxide on APL --- p.19 / Chapter 1.5.3.4 --- In vitro and in vivo Study of Arsenic Trioxide (As2O3) in treating APL --- p.19 / Chapter 1.5.3.5 --- Common Side Effects of Arsenic Trioxide (As2O3) on APL --- p.21 / Chapter 1.5.4 --- Anti-cancer effect of Arsenic Trioxide on other cancers --- p.23 / Chapter 1.6 --- Aim of Study --- p.24 / Chapter Chapter 2 --- Materials and Methods --- p.26 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.27 / Chapter 2.1.1.1 --- Cell Lines --- p.27 / Chapter 2.1.1.2 --- Culture Medium --- p.27 / Chapter 2.1.2 --- Chemicals --- p.28 / Chapter 2.1.3 --- Buffers and Reagents --- p.29 / Chapter 2.1.4 --- Reagents for MTT Assay --- p.30 / Chapter 2.1.5 --- Reagents for DNA Fragmentation --- p.31 / Chapter 2.1.5.1 --- Reagents for DNA Extraction --- p.31 / Chapter 2.1.5.2 --- Reagents for Gel Electrophoresis --- p.31 / Chapter 2.1.6 --- Reagents for Western Blotting --- p.32 / Chapter 2.1.6.1 --- Reagents for Protein Extraction --- p.32 / Chapter 2.1.6.2 --- Reagents for SDS-PAGE --- p.33 / Chapter 2.1.7 --- Reagents for Flow Cytometry --- p.36 / Chapter 2.1.8 --- In Vivo Study --- p.37 / Chapter 2.2 --- Methods --- p.38 / Chapter 2.2.1 --- Cell Treatment --- p.38 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.38 / Chapter 2.2.3 --- MTT Assay --- p.38 / Chapter 2.2.4 --- Detection of DNA Fragmentation --- p.39 / Chapter 2.2.5 --- Flow Cytometry --- p.40 / Chapter 2.2.5.1 --- Detection of Cell Cycle Pattern with PI --- p.40 / Chapter 2.2.5.2 --- Detection of Apoptosis with Annexin V-PI --- p.40 / Chapter 2.2.6 --- Western Blot Analysis --- p.41 / Chapter 2.2.6.1 --- Protein Extraction --- p.41 / Chapter 2.2.6.2 --- Protein Concentration Determination --- p.41 / Chapter 2.2.6.3 --- Western Blotting --- p.42 / Chapter 2.2.7 --- In Vivo Study --- p.44 / Chapter 2.2.7.1 --- Animal Model --- p.44 / Chapter 2.2.7.2 --- Treatment Schedule --- p.44 / Chapter 2.2.7.3 --- Toxicity of Arsenic Trioxide --- p.45 / Chapter Chapter 3 --- Anti-Proliferation Effect of As2O3 on MDA-MB-231 cells --- p.47 / Chapter 3.1 --- Study the Anti-proliferation Effect of As2O3 on MDA-MB-231 Cells by MTT Assay --- p.48 / Chapter 3.2 --- Comparsion Anti-proliferation Effect of AS2O3 on MDA-MB-231 Cells to that of Tamoxifen --- p.50 / Chapter 3.3 --- "Study Toxicity of AS2O3 on Normal Breast Cells Line, 184B5" --- p.52 / Chapter 3.4 --- Summary --- p.54 / Chapter Chapter 4 --- Mechanism of Growth Inhibition Effect of As2O3 on MDA-MB-231 cells --- p.56 / Chapter 4.1 --- Cell Cycle Analysis of As2O3 Treated MDA-MB-231 Cells --- p.57 / Chapter 4.2 --- Detection of DNA Fragmentation --- p.60 / Chapter 4.3 --- Detection of Apoptosis Induced by AS2O3 on MDA-MB-231 Cells by Flow Cytometry --- p.62 / Chapter 4.4 --- Regulation of Apoptotic Related Protein by As2O3 on MDA-MB-231 Cells --- p.64 / Chapter 4.4.1 --- Expression Level of Bcl-2 and Bax Protein --- p.66 / Chapter 4.4.2 --- Expression Level of Cytochrome C --- p.69 / Chapter 4.4.3 --- Expression Level of Caspase9 --- p.71 / Chapter 4.4.4 --- Expression Level of FasL --- p.73 / Chapter 4.4.5 --- Expression Level of Caspase8 --- p.75 / Chapter 4.4.6 --- Expression Level of Caspase3 --- p.77 / Chapter 4.4.7 --- Expression Level of Poly (ADP-ribose) Polymerase (PARP) --- p.79 / Chapter 4.4.8 --- Expression Level of p53 --- p.81 / Chapter 4.5 --- Regulation of Cell Cycle Related Protein by AS2O3 on MDA-MB-231 Cells --- p.83 / Chapter 4.5.1 --- Expression Level of Cyclin B --- p.84 / Chapter 4.5.2 --- Expression Level of Cyclin E --- p.86 / Chapter 4.6 --- Summary --- p.88 / Chapter Chapter 5 --- In Vivo Study of Anti-tumor Effect of As2O3 --- p.89 / Chapter 5.1 --- Anti-tumor Effect of AS2O3 on Tumor Bearing Nude Mice --- p.90 / Chapter 5.2 --- Toxic Effect of AS2O3 on Normal Tissues --- p.93 / Chapter 5.3 --- Summary --- p.98 / Chapter Chapter 6 --- Discussion --- p.99 / Chapter 6.1 --- Anti-tumor Effect of AS2O3 on Breast Cancer --- p.100 / Chapter 6.2 --- Induction of Apoptosis and Cell Cycle arrest by AS2O3 --- p.101 / Chapter 6.3 --- Side Effect of AS2O3 on Breast Cancer Treatment --- p.103 / Chapter Chapter 7 --- Future Perspectives --- p.105 / Chapter 7.1 --- Future Perspectives --- p.106 / References --- p.108

Li, Wing Yan Kate. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 130-154). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / Abstract (Chinese Version) --- p.vi / Table of Contents --- p.viii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1 --- General Introduction / Chapter 1.1. --- Overview of cancer --- p.1 / Chapter 1.2. --- Apoptosis and cancer --- p.4 / Chapter 1.3. --- Roles and regulation of caspase-dependent apoptosis --- p.7 / Chapter 1.3.1. --- Extrinsic death receptor pathway --- p.8 / Chapter i. --- TNFR1 and TNFa --- p.13 / Chapter ii. --- CD95/Fas and CD95 Ligand/FasL --- p.14 / Chapter iii. --- "TRAIL-R1(DR4), TRAIL-R2 (DR5) and TRAIL" --- p.14 / Chapter 1.3.2. --- Intrinsic mitochondrial pathway --- p.16 / Chapter i. --- Bcl-2 family of proteins --- p.17 / Chapter ii. --- Reactive Oxygen Species (ROS) --- p.19 / Chapter 1.4. --- Phytochemicals from Traditional Chinese Medicine (TCM) as a source of new therapeutics --- p.22 / Chapter 1.5. --- Biological effects of baicalein --- p.25 / Chapter 1.5.1 --- Roles of baicalein as a lipoxygenase inhibitor --- p.28 / Chapter 1.5.2 --- Dual roles of baicalein as an antioxidant and prooxidant --- p.28 / Chapter 1.5.3 --- "Roles of baicalein as an anti-carcinogenic, anti-proliferative and anti-metastatic agent" --- p.29 / Chapter 1.6. --- Aims of current study --- p.30 / Chapter Chapter 2 --- Effects of Baicalein on Growth and Survival of Human Cancer Cells / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Cell culture --- p.35 / Chapter 2.2.2 --- Measurement of growth and survival of various cell lines --- p.36 / Chapter 2.2.3 --- Statistical analysis --- p.37 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Baicalein retards the growth and survival of human melanoma A375 and colorectal carcinoma Caco-2 --- p.37 / Chapter 2.3.2 --- Baicalein reduces the growth and survival of melanoma A375 but not in normal skin fibroblast Hs68 cells --- p.40 / Chapter 2.4 --- Discussion --- p.42 / Chapter Chapter 3 --- Effects of Baicalein on Cell Cycle and the Apoptosis in Human Melanoma A375 Cells / Chapter 3.1 --- Introduction --- p.44 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Determination of cell cycle changes and quantification of apoptosis --- p.51 / Chapter 3.2.2 --- Immunoblotting --- p.52 / Chapter 3.2.3 --- Inhibition of caspase-8 by caspase-8 inhibitor --- p.54 / Chapter 3.2.4 --- Fluorometric measurement of caspase-3 activity --- p.54 / Chapter 3.2.5 --- Statistical analysis --- p.55 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Baicalein induces S-phase arrest in cell cycle and triggers apoptosis --- p.55 / Chapter 3.3.2 --- Baicalein induces proteolytic inactivation of PARP and activation of caspases --- p.59 / Chapter 3.3.3 --- Caspase-8 is the major initiator caspase eliciting the baicalein-induced apoptosis --- p.62 / Chapter 3.4 --- Discussion --- p.67 / Chapter Chapter 4 --- Effects of Baicalein on the Extrinsic Apoptotic Pathways in Human Melanoma A375 Cells / Chapter 4.1 --- Introduction --- p.72 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Immunoblotting --- p.75 / Chapter 4.2.2 --- Determination of sub-lethal dose of exogenous TRAIL --- p.76 / Chapter 4.2.3 --- Determination of the combinatory effect of exogenous TRAIL and baicalein --- p.76 / Chapter 4.2.4 --- Statistical analysis --- p.77 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Baicalein upregulates the expressions of death receptor 4 (DR4) and death receptor 5 (DR5) --- p.77 / Chapter 4.3.2 --- Baicalein sensitizes the melanoma cells to sub-lethal dose of exogenous TRAIL --- p.80 / Chapter 4.4 --- Discussion --- p.84 / Chapter Chapter 5 --- Effects of Baicalein on the Extrinsic Apoptotic Pathways in Human Melanoma A375 Cells Cancer Cells / Chapter 5.1 --- Introduction --- p.88 / Chapter 5.2 --- Materials and Methods / Chapter 5.2.1 --- Analysis of mitochondrial membrane potential --- p.94 / Chapter 5.2.2 --- Fractionation of cell lysates into cytosolic and mitochondrial fractions for immunoblotting --- p.95 / Chapter 5.2.3 --- Immunoblotting --- p.95 / Chapter 5.2.4 --- Determination of cellular reactive oxygen species (ROS) production --- p.96 / Chapter 5.2.5 --- Verification of ROS generation via the addition of Trolox´ёØ --- p.96 / Chapter 5.2.6 --- Statistical analysis --- p.97 / Chapter 5.3 --- Results / Chapter 5.3.1 --- Baicalein induces mitochondrial membrane depolarization --- p.97 / Chapter 5.3.2 --- Cytochrome c is released in the baicalein-induced mitochondrial membrane depolarization --- p.100 / Chapter 5.3.3 --- Baicalein does not elicit the intrinsic apoptotic pathway via modulation of some better-characterized Bcl-2 family proteins in A375 cells --- p.102 / Chapter 5.3.4 --- Baicalein induces ROS production --- p.105 / Chapter 5.3.5 --- Baicalein induces mitochondrial permeabilization via ROS-mediated mechanisms --- p.108 / Chapter 5.4 --- Discussion --- p.112 / Chapter Chapter 6 --- General Discussion --- p.119 / References --- p.130

Cheung, Kwok Kuen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 226-263). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

O tratamento farmacológico em idosos representa grande desafio, principalmente relacionado às alterações no metabolismo, que resultam em modificações farmacocinéticas e farmacodinâmicas, aumentando a sensibilidade a muitos fármacos. Associado ao uso extensivo de medicamentos e a presença de comorbidades, o risco de interações medicamentosas aumenta substancialmente nesta população. Essa problemática destaca-se em idosos institucionalizados em hospitais psiquiátricos. Desse modo, esta pesquisa teve como objetivo geral analisar e classificar as interações medicamentosas (IM) potenciais do tipo fármaco-fármaco, em idosos institucionalizados em unidades de internação e lares abrigados de Centro de Atenção Integral à Saúde. Trata-se de estudo de delineamento não experimental, descritivo correlacional e transversal. O estudo foi realizado a partir de dados secundários contidos nos prontuários dos idosos com transtornos psiquiátricos e neuropsiquiátricos que foram internados em um Centro de Atenção Integral à Saúde estadual nos anos de 2005 a 2014. Foram verificados todos os medicamentos prescritos na admissão do paciente, na mediana da internação e na última prescrição. Para coleta de dados foi utilizado um instrumento adaptado de Reis (2009). Para análise dos dados, foi utilizada a análise descritiva e a regressão logística múltipla bruta e ajustada. A maioria dos idosos era do sexo feminino (61,7%), com idade de 60 a 69 anos (54,3%), com diagnóstico psiquiátrico e clínico concomitantes (72,3%) e em uso de 5 a 10 medicamentos (68,1%). Os fármacos utilizados para o Sistema Nervoso foram os mais prescritos. Verificou-se que 67,0%, 74,5% e 80,8% dos pacientes apresentaram IM potenciais na admissão, mediana da internação e última prescrição, respectivamente. Foi detectado um total de 1064 IM potenciais nos três momentos investigados e constatou-se a média de aproximadamente cinco IM potenciais nas prescrições por paciente. Houve elevada porcentagem de IM potenciais graves e contraindicadas. No modelo de regressão logística, realizado para identificar os preditores de IM, no momento da admissão dos idosos, contribuíram de forma significativa a idade de 60 a 69 anos (OR=7,9), a polifarmácia (OR=16,6) e o uso de medicamentos para o aparelho cardiovascular (OR=11,3) e para o aparelho respiratório (OR=16,4). Na mediana da internação, foram preditores de IM potenciais o uso de medicamentos que atuam no sistema nervoso (OR=7,4) e polifarmácia (OR=4,9). Na última prescrição, apenas a polifarmácia foi preditor para ocorrência de IM potencial (OR=30,1), indicando que a segurança do paciente no uso de medicamentos permaneceu comprometida. Os resultados desta investigação mostraram o comprometimento da segurança dos idosos no que se refere ao potencial para IM. Estratégias como a integração da equipe multidisciplinar, a reconciliação medicamentosa e a atuação do enfermeiro na avaliação do processo medicamentoso do idoso são necessárias para o cuidado seguro e de qualidade / Pharmacological treatment in elderly represents a great challenge, mainly related to changes in metabolism that result in pharmacokinetic and pharmacodynamic changes, increasing sensitivity to many medications. Associated with the extensive use of medication and the presence of comorbidities, the risk of drug interactions increases substantially in this population. This problem stands out in institutionalized elderly in psychiatric hospitals. Thus, this study aimed to analyze and classify potential drug-drug interactions (DDI), in institutionalized elderly in inpatient units and sheltered homes in a Center of Comprehensive Health Care. It\'s a not experimental, descriptive correlational and transversal study. The study was conducted from secondary data in the medical records of elderly patients with psychiatric and neuropsychiatric disorders that have been admitted to a Center of Comprehensive Health Care from the years 2005 to 2014. We checked all drugs prescription at the admission of hospitalization, at median of stay and at the last prescription. For data collection was used an adapted instrument of Reis (2009). For data analysis, we used the descriptive analysis and multiple logistic regression. The majority seniors were female (61.7%) aged from 60 to 69 years (54.3%), with psychiatric and clinical diagnosis concomitant (72.3%) and in use of 5 to 10 drugs (68.1%). The most prescribed drugs were those used to the nervous system. It was found that 67.0%, 74.5% and 80.8% of patients had potential DDI at admission, at median of stay and at the last prescription, respectively. It was detected a total of 1064 potential IM in the three investigated moments and found to average approximately five potential DI requirements for the patient. There was a high percentage of serious and contraindicated potencial DDI. In the logistic regression model, performed to identify predictors of DDI at the time of admission of the elderly, the age 60-69 years (OR=7.9), polypharmacy (OR=16.6) and use of drugs for the cardiovascular system (OR=11.3) and the respiratory system (OR=16.4) contributed significantly to potential DDI. At the median of stay were predictors of potential DDI the use of drugs that operate on the nervous system (OR=7.4) and polypharmacy (OR=4.9). In the last prescription, only polypharmacy was predictor of potential DDI occurrence (OR=30.1), indicating that patient safety in medication use remained compromised. The results of this investigation showed the impairment of the elderly safety regarding potential for drug interactions. Strategies such as the integration of the multidisciplinary team, the medication reconciliation and the work of nurses in assessing the elderly drug\'s process are necessary for a safe and a qualitied care

Introducão. A perda da resposta aos mecanismos que regulam o crescimento da população celular em tecidos normais, as alterações em níveis genéticos e epigenéticos, induzem a carcinogênese. Durante o crescimento, seja tumor primário ou metástase, ge- ralmente há uma fase avascular, caracterizada por tumores pequenos onde os recursos necessário para proliferação da população são oriundos dos tecidos adjacentes. Ade- mais, um aspecto marcante é a heterogeneidade intratumoral, que é desencadeada por fatores intrínseco e ambientais às células, desempenhando papeis importante no cresci- mento e resposta ao tratamento. Métodos. Foi proposto um modelo estocástico espacial em termos de Cadeia de Markov para simular a dinâmica de populações de um câncer in situ em fase avascular e o tratamento quimioterapico. As transições de estado das célu- las consideradas foram duplicação, migração, morte, geração de variação na população e quiescência. Assumimos que as taxas de duplicação, migração e morte, dependem do tamanho da população, onde a taxa de duplicação diminui conforme a população au- menta e as taxas de migração e morte aumentam conforme a população cresce. As taxas dependente do tamanho da população foram descritos em termos de funções sigmóidais onde a suavidade da transição entre os valores mínimos e máximos das taxas foi dado pelo parâmetro k, o tamanho da população na qual a taxa de duplicação cai na metade do valor máximo foi dado por N e a taxa de duplicação na qual as taxa de migração/morte atingem a metade do valor máximo por 0\'. A heterogeneidade do tumor foi representada como um conjunto de duas subpopulações (1) mais proliferativa e (2) mais migratória. Em seguida, foi aclopado um modelo de difusão de drogas e um modelo farmacodinâ- mico para avaliar a repopulação no tratamento com uma única droga. Resultados. As simulações acerca do crescimento tumoral mostram que a população atingiu a densidade de saturação de acordo com os valores do parâmetro k. A densidade das variantes prol iferativas e migratórias não foram diretamente relacionadas a taxa de geração de variação, mas sim pelos parâmetros N e a. Para determinado conjunto de valores atribuídos aos parâmetros k e N,espacialmente observou-se crescimento difuso da população. No que tange ao tratamento, houve emergência da resistência, todavia observou-se padrões diferente na repopulação de acordo com os valores da taxa de geração de variação, k e N. Espacialmente, observa-se que as células resistentes vão ao gradiente droga. O modelo no estágio atual consegue recuperar qualitativamente algumas evidências experimentais atreladas coexistência de duas subpopulações onde uma cresce mais do que a outra, crescimento difuso da população e migração da população resistente para o gradiente de droga / Introduction. The loss of response to the mechanism which regulates the growth of cell population in normal tissue, the changes at both genetic and epigenetic levei lead to carcinogenesis. During the growth of primary tumor or metastasis often there is an avascular phase which is featured by small tumor where the resource necessary to proli- feration nd survival come from surrounding tissues. Furthermore, a remarkable aspect of carcinogenesis is intratumoral heterogeneity which is fueled by both intrinsic and environ- mental facto r, playing a crucial role in the growth and treatment response. Methods Here we propose a stochastic spatial model in term of Markov Chain to simulate the population dynamic of in situ cancer in avascular phase and the chemotherapeutic treatment. The cells state transition considered was duplication, migration, death, generation of new variants in population and quiescence. We are assuming that duplication, migration and death rates depend on population size, where duplication rates decrease as long as population increase but both migration and death rates, in it turn increase. Theses population size dependent rates were described in terms of sigmoidal function where the smoothness of transition between maximal e minimum values was indicated by k, the size of population at which duplication rates falls on half of it maximal value was indicated by Nand the duplication rate where both migration and death rates goes to half of it maximal value was indicated by a. The heterogeneity was featured as a set of a (1) highly proliferative and (2) highly migratory cells. In doing so, we coupled a drug diffusion drug and a phar- macodynamical model in order to evaluate the repopulation in the single drug treatment. The simulations about tumor growth showed that population reached a saturation density accordingly with values of the parameter k.The density of both proliferative and migratory variants was not directed related to the generation of variants rates but by the parameter Nand a. To some specific set of assigned values to the parameters k and N,spatially was observed that diffuse growth of population. Concerning to the treatment, there was resistance emergence, h oweve r, we observed differents patterns in the repopulation ac- cordingly the generation of variants rates, k and N.Spatially, we observed the resistant cells goes to drug gradient. The model, currently, has recovered qualitatively some ex- perimental evidence linked to the co-existence of two subpopulations where one of them grow more than other, diffuse growth of population and migration of resistant population to drug gradient

Dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Master of Science in Medicine. Johannesburg, 2014 / Background: The causes of renal dysfunction in children treated for childhood cancers are multifactorial and clinical manifestations of dysfunction include hypertension, proteinuria and varying degrees of renal insufficiency. This study aimed to determine the different residual effects of cancer therapy on the renal system and factors associated with the residual effects in children treated for childhood cancers. Patients and Methods: The study was a descriptive cross sectional study that assessed 130 children, between the age of 1 and 18 years, who had completed treatment at Charlotte Maxeke Johannesburg Academic Hospital and were being followed up at the paediatric oncology clinic of the hospital. Results: After a median follow-up post treatment of 2 years, the various manifestations of renal dysfunction identified in the survivors included; decreased GFR, hypomagnesaemia, hypophosphataemia, proteinuria, haematuria and hypertension. In total, 34 survivors (26.15%) had at least one manifestation of renal dysfunction after completing treatment. The most prevalent manifestation of renal dysfunction detected was decreased GFR (17.69%). Hypomagnesaemia and hypophosphataemia were present in 8 (6.15%) and 6 (4.62%) of the survivors respectively. Patients who had renal dysfunction pre-treatment were three times more likely to have renal dysfunction post-treatment. Ifosfamide, Carboplatinum, and nephrectomy were significantly associated with a reduction in GFR Conclusion: A significant number of the survivors had a decreased GFR while some of them had hypomagnesaemia and hypophosphataemia. There was a strong association between pre-treatment and post-treatment renal dysfunction. These findings are very important in terms of decision making for individual patients with respect to selecting treatment modalities and dosages and also with respect to instituting nephro-protective measures to avoid further damage to the kidneys during and after treatment.