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Immunomodulatory effects of yun zhi and danshen capsules in healthy subjects: a randomized, double-blind, placebo-controlled crossover study.January 2003 (has links)
Tse Pui Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves [191]-216). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABBREVIATIONS --- p.III / ABSTRACT --- p.VIII / 摘要 --- p.X / PUBLICATIONS --- p.XII / TABLE OF CONTENTS --- p.XIII / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Human Immune System and Cancer --- p.1 / Chapter 1.1.1 --- Brief Introduction of the Human Immune System --- p.1 / Chapter 1.1.2 --- Prevalence of Cancer in Hong Kong --- p.4 / Chapter 1.1.3 --- The Role of the Immune System in Tumorigenesis --- p.4 / Chapter 1.1.4 --- Cancer Treatment --- p.5 / Chapter 1.1.5 --- Cancer Prevention --- p.5 / Chapter 1.2 --- Mushroom Polysaccharides --- p.6 / Chapter 1.2.1 --- General Aspects of Mushroom Polysaccharides --- p.6 / Chapter 1.2.2 --- Structure of Mushroom Polysaccharides --- p.9 / Chapter 1.2.2.1 --- Beta (P)-D-glucans --- p.9 / Chapter 1.2.2.2 --- Heteroglucans and Protein-bound Polysaccharides --- p.10 / Chapter 1.2.2.3 --- Structure-Function Interactions of Polysaccharides --- p.12 / Chapter 1.2.3 --- Molecular Interactions of Polysaccharides --- p.14 / Chapter 1.2.4 --- Biological Activities of Polysaccharides --- p.15 / Chapter 1.2.4.1 --- Anti-tumor Activities of Polysaccharides --- p.15 / Chapter 1.2.4.2 --- Immunomodulatory Activities of Polysaccharides --- p.16 / Chapter 1.3 --- Yun Zhi (Coriolus versicolor) --- p.17 / Chapter 1.3.1 --- General Features of Yun Zhi --- p.17 / Chapter 1.3.2 --- Traditional Uses of Yun Zhi --- p.20 / Chapter 1.3.3 --- Active Ingredients of Yun Zhi --- p.20 / Chapter 1.3.3.1 --- "Origin, Properties and Composition of PSK" --- p.21 / Chapter 1.3.3.2 --- "Origin, Properties and Composition of PSP" --- p.22 / Chapter 1.3.4 --- Pharmacological Actions of PSP and PSK --- p.25 / Chapter 1.3.4.1 --- Immunomodulatory Activities --- p.25 / Chapter 1.3.4.2 --- Anti-tumor Activities --- p.32 / Chapter 1.3.4.2 --- Antiviral and Antimicrobial Activities --- p.35 / Chapter 1.3.4.3 --- Antioxidant Activities --- p.36 / Chapter 1.3.5 --- Human Clinical Studies on Yun Zhi --- p.36 / Chapter 1.3.6 --- Toxicology of Yun Zhi --- p.42 / Chapter 1.4 --- Danshen (Salvia miltiorrhiza) --- p.43 / Chapter 1.4.1 --- General Features of Danshen --- p.43 / Chapter 1.4.2 --- Traditional Uses of Danshen --- p.46 / Chapter 1.4.3 --- Active Ingredients of Danshen --- p.47 / Chapter 1.4.4 --- Pharmacological Actions of Danshen --- p.50 / Chapter 1.4.4.1 --- Cardiovascular Effects --- p.50 / Chapter 1.4.4.2 --- Scavenging Effects on Free Radicals --- p.52 / Chapter 1.4.4.3 --- Hepatoprotective Effects --- p.54 / Chapter 1.4.4.4 --- Anti-tumor Effects --- p.56 / Chapter 1.4.4.5 --- Renal Protective Effects --- p.56 / Chapter 1.4.5 --- Human Clinical Studies --- p.57 / Chapter 1.4.6 --- Toxicity of Danshen --- p.59 / Chapter 1.5 --- Aims and Scopes of This Investigation --- p.60 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Normal Subjects --- p.62 / Chapter 2.1.1 --- Inclusion and Exclusion Criteria of Recruitment --- p.62 / Chapter 2.1.2 --- Study Design and Procedure --- p.63 / Chapter 2.1.3 --- Treatment and Blinding --- p.65 / Chapter 2.1.4 --- Blood Sampling --- p.66 / Chapter 2.1.5 --- Blood Processing for Assessment of Immunological Functions --- p.67 / Chapter 2.2 --- Materials --- p.69 / Chapter 2.2.1 --- Endotoxin Assay --- p.69 / Chapter 2.2.2 --- Reagents for Whole Blood Assay --- p.69 / Chapter 2.2.2.1 --- Plain RPMI 1640 Medium --- p.69 / Chapter 2.2.2.2 --- Phosphate-Buffered Saline (PBS) --- p.69 / Chapter 2.2.2.3 --- Mitogens --- p.70 / Chapter 2.2.3 --- Reagents for Total RNA Extraction --- p.70 / Chapter 2.2.3.1 --- Ficoll-Paque Density Gradient Solution --- p.70 / Chapter 2.2.3.2 --- RNA Extraction Kit --- p.70 / Chapter 2.2.3.3 --- RNase-Free DNase Set --- p.71 / Chapter 2.2.3.4 --- β-Mercaptoethanol (β-ME) Solution --- p.71 / Chapter 2.2.4 --- Reagents for Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.71 / Chapter 2.2.4.1 --- MultiTEST IMK Kit with TruCOUNT Tubes --- p.71 / Chapter 2.2.4.2 --- FACSFlo´wёØ Sheath Fluid --- p.74 / Chapter 2.2.4.3 --- CaliBRITE 3 and APC Beads --- p.74 / Chapter 2.2.5 --- Immunoassay Kits for Measuring Cytokines Level --- p.75 / Chapter 2.2.5.1 --- Enzyme-linked Immunosorbent Assay (ELISA) Kits of Cytokines --- p.75 / Chapter 2.2.5.2 --- Human Thl/Th2 Cytokine Cytometric Bead Array (CBA) Kit-II --- p.75 / Chapter 2.2.6 --- Reagents and Buffers for Gel Electrophoresis --- p.78 / Chapter 2.2.6.1 --- Ethidium Bromide (EtBr) --- p.78 / Chapter 2.2.6.2 --- Gel Loading Solution (5X) --- p.78 / Chapter 2.2.6.3 --- Tris-Acetate-EDTA (TAE) Buffer --- p.78 / Chapter 2.2.6.4 --- Agarose Gel --- p.78 / Chapter 2.2.6.5 --- 100 base pair DNA Ladder --- p.79 / Chapter 2.2.7 --- Kits and Reagents for Messenger RNA (mRNA) Expression Array --- p.79 / Chapter 2.2.7.1 --- Human Inflammatory Cytokine/Receptor GEArraýёØ Q Series Kit --- p.79 / Chapter 2.2.7.2 --- Deoxynucleoside Triphosphates (dNTPs) --- p.84 / Chapter 2.2.7.3 --- Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLVRT) --- p.84 / Chapter 2.2.7.4 --- Rnasin Ribonuclease Inhibitor --- p.84 / Chapter 2.2.7.5 --- Biotin-16-2'-deoxy-uridine-5'-triphosphate (Biotin-16-dUTP) --- p.85 / Chapter 2.2.7.6 --- Salmon Sperm DNA Solution --- p.85 / Chapter 2.2.7.7 --- 100 % Sodium Dodecyl Sulfate (SDS) Solution --- p.86 / Chapter 2.2.7.8 --- 20X SSC --- p.86 / Chapter 2.2.7.9 --- ECL Films (Hyperfilm 226}0ёØ ECL 226}0ёØ) --- p.86 / Chapter 2.3 --- Methods / Chapter 2.3.1 --- Endotoxin Assay --- p.87 / Chapter 2.3.2 --- Whole Blood Assay (WBA) --- p.88 / Chapter 2.3.3 --- Isolation and Preparation of Plasma and Peripheral Blood Mononuclear Cells (PBMC) from EDTA Blood --- p.88 / Chapter 2.3.4 --- Total RNA extraction --- p.89 / Chapter 2.3.5 --- Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.90 / Chapter 2.3.6 --- Immunoassays of Plasma Samples or Culture Supernatant in WBA --- p.92 / Chapter 2.3.6.1 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.92 / Chapter 2.3.6.2 --- Human Thl/Th2 Cytokine Cytometric Bead Assay (CBA) --- p.93 / Chapter 2.3.7 --- mRNA Expression Study --- p.94 / Chapter 2.3.7.1 --- Agarose Gel Electrophoresis --- p.94 / Chapter 2.3.7.2 --- cDNA Expression Array Analysis --- p.95 / Chapter 2.3.8 --- Statistical Analysis --- p.96 / Chapter CHAPTER 3: --- ENDOTOXIN LEVEL OF YUN ZHI-DANSHEN CAPSULES & SAFETY MEASURES ON STUDY POPULATION IN THE CLINICAL TRIAL / Chapter 3.1 --- Introduction --- p.98 / Chapter 3.2 --- Results --- p.101 / Chapter 3.2.1 --- Endotoxin Level of the Yun Zhi and Danshen Active Capsule --- p.101 / Chapter 3.2.2 --- Study Population --- p.103 / Chapter 3.2.3 --- Dropout Cases --- p.103 / Chapter 3.2.4 --- Safety Parameters --- p.104 / Chapter 3.2.5 --- Compliance Rates --- p.104 / Chapter 3.3 --- Discussion --- p.109 / Chapter CHAPTER 4: --- FLOW CYTOMETRIC ANALYSIS OF T/B/NK CELL RATIOS OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES / Chapter 4.1 --- Introduction --- p.112 / Chapter 4.2 --- Results --- p.118 / Chapter 4.2.1 --- The Percentage and Absolute Count of T Lymphocytes (CD3+) --- p.118 / Chapter 4.2.2 --- The Percentage and Absolute Count of T Helper (TH) Lymphocytes (CD3+ CD4+) --- p.121 / Chapter 4.2.3 --- The Percentage and Absolute Count of Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocytes (CD3+ CD8+) --- p.124 / Chapter 4.2.4 --- The Ratio of T Helper Lymphocytes (CD3+ CD4+) and Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocyes (CD3+ CD8+) --- p.127 / Chapter 4.2.5 --- The Percentage and Absolute Count of B Lymphocytes (CD19+) --- p.129 / Chapter 4.2.6 --- The Percentage and Absolute Count of NK Lymphocytes (CD3- CD 16+ and/or CD56+) --- p.132 / Chapter 4.2.7 --- The Absolute Count of Lymphocytes (CD45+) --- p.135 / Chapter 4.3 --- Discussion --- p.138 / Chapter CHAPTER 5: --- PLASMA CONCENTRATION OF SOLUBLE CYTOKINE RECEPTOR AND EX VIVO CYTOKINE PRODUCTION OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES / Chapter 5.1 --- Introduction --- p.142 / Chapter 5.2 --- Results --- p.147 / Chapter 5.2.1 --- Plasma Concentration of Soluble IL-2 Receptor --- p.147 / Chapter 5.2.2 --- Ex vivo Cytokine Production --- p.147 / Chapter 5.2.3 --- Mitogen Induced IL-6 Production --- p.150 / Chapter 5.2.4 --- Mitogen Induced IFN- γ Production --- p.150 / Chapter 5.2.5 --- Mitogen Induced TNF- a Production --- p.153 / Chapter 5.2.6 --- Mitogen Induced IL-10 Production --- p.153 / Chapter 5.3 --- Discussion --- p.156 / Chapter CHAPTER 6: --- "GENE EXPRESSION OF CYTOKINES, CHEMOKINES AND RECEPTORS OF PBMC OF HEALTHY SUBJECTS TAKING YUN ZHI- DANSHEN CAPSULES" / Chapter 6.1 --- Introduction --- p.162 / Chapter 6.2 --- Results --- p.165 / Chapter 6.2.1 --- Gene Expression of IL-2 Receptor β chain --- p.165 / Chapter 6.2.2 --- Gene Expression of IL-2 Receptor γ chain --- p.165 / Chapter 6.2.3 --- Gene Expression of IL-6 Receptor --- p.166 / Chapter 6.2.4 --- "Gene Expression of Other Cytokines, Chemokines and Receptors" --- p.169 / Chapter 6.3 --- Discussion --- p.172 / Chapter CHAPTER 7: --- CONCLUDING REMARKS AND FUTURE / PERSPECTIVES --- p.176 / APPENDICES --- p.184 / REFERENCES --- p.192
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