Lee Ka Wai Sharon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 227-260). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABBREVIATIONS --- p.III / ABSTRACT --- p.VI / 摘要 --- p.XI / CONFERENCE PUBLICATIONS --- p.XVII / TABLE OF CONTENTS --- p.XVIII / Chapter Part I - --- General Introduction / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- The Search of Immunomodulatory Agents --- p.1 / Chapter 1.2. --- Cordyceps sinensis (Dong Cong Xia Cao) as an Immunomodulatory Agent --- p.2 / Chapter 1.2.1. --- General Aspects --- p.2 / Chapter 1.2.2. --- Evidence from the Traditional Chinese Medicine Concepts --- p.2 / Chapter 1.2.3. --- Evidence from the Traditional Chinese Medicine Classics --- p.4 / Chapter 1.2.4. --- Evidence from the Modern Research Literature --- p.4 / Chapter 1.2.4.1. --- lmmunomodulation --- p.4 / Chapter 1.2.4.2. --- Anti-tumor Effects --- p.7 / Chapter 1.2.4.3. --- Other Activities Related to the Immune System --- p.8 / Chapter 1.2.4.4. --- Potential Active Ingredients: Cordycepin and Polysaccharides --- p.8 / Chapter 1.2.5. --- Prescription and Usage: Single Vs Concocted --- p.11 / Chapter 1.2.5.1. --- Single Form as an Immunoactivating Agent --- p.11 / Chapter 1.2.5.2. --- Concocted as an Anti-asthmatic Agent --- p.12 / Chapter 1.3. --- Our Hypothesis and Rationale --- p.13 / Chapter Chapter 2: --- Experimental Design --- p.24 / Chapter 2.1. --- General Aspects of the Human Immune System --- p.24 / Chapter 2.2. --- Designing the In vitro Study on Cell-mediated Immunity --- p.24 / Chapter 2.2.1. --- T Cells --- p.25 / Chapter 2.2.2. --- Macrophages --- p.26 / Chapter 2.3. --- Designing the In vitro and In vivo Study of Anti-tumor Activities --- p.29 / Chapter 2.3.1. --- Tumor Biology --- p.29 / Chapter 2.3.2. --- Tumor and Immunity --- p.29 / Chapter 2.3.2.1. --- T-Cell-Mediated Cytolysis (Tc Cells) --- p.30 / Chapter 2.3.2.2. --- Delayed-Type Hypersensitivity (TDth Cells) --- p.30 / Chapter 2.3.2.3. --- Natural Killer (NK) Cells --- p.30 / Chapter 2.3.2.4. --- Lymphokine-Activated Killer (LAK) Cells --- p.31 / Chapter 2.3.2.5. --- Antibody-Dependent Cell-Mediated Cytotoxic (ADCC) Cells --- p.31 / Chapter 2.3.2.6. --- Activated Macrophages (AMΦ) --- p.31 / Chapter 2.3.3. --- Mechanism of Tumor Engulfment --- p.32 / Chapter 2.3.4. --- The Experimental Plan --- p.33 / Chapter 2.4. --- Designing the In vitro Study and Clinical Trials on Anti-asthmatic Activities --- p.36 / Chapter Part II - --- Methodology / Chapter Chapter 3: --- Materials and Methods / Chapter 3.1. --- List of Materials and Their Origin --- p.39 / Chapter 3.1.1. --- Traditional Chinese Medicine --- p.39 / Chapter 3.1.2. --- Cells for In vitro Experiments --- p.39 / Chapter 3.1.3. --- Mice for In vivo Experiments --- p.40 / Chapter 3.1.4. --- "Medium, Buffer, Supplements and Reagents for Cell Culture" --- p.40 / Chapter 3.1.5. --- Dye for Cellular Staining --- p.40 / Chapter 3.1.6. --- Cell Mitogens and Activator --- p.41 / Chapter 3.1.7. --- Reagents for Flow Cytometric Analysis --- p.41 / Chapter 3.1.8. --- Reagent Kits --- p.41 / Chapter 3.1.9. --- ELISA Kits --- p.42 / Chapter 3.1.10. --- Antibodies --- p.43 / Chapter 3.1.11. --- Reagents for RNA Extraction --- p.44 / Chapter 3.1.12. --- Reagents for Gel Electrophoresis --- p.44 / Chapter 3.1.13. --- Reagents for cDNA Expression Array --- p.44 / Chapter 3.1.14. --- Other Reagents --- p.45 / Chapter 3.1.15. --- Special Equipment and Apparatus --- p.45 / Chapter 3.2. --- Details of Materials --- p.46 / Chapter 3.2.1. --- Traditional Chinese Medicine --- p.46 / Chapter 3.2.1.1. --- Natural Cordyceps sinensis --- p.46 / Chapter 3.2.1.2. --- HERBSnSENSEŚёØ Cordyceps --- p.46 / Chapter 3.2.1.3. --- Wheeze-Relief Formula --- p.46 / Chapter 3.2.2. --- "Media, Supplements and Reagents for Cell Culture" --- p.47 / Chapter 3.2.2.1. --- Cell Culture Media --- p.47 / Chapter 3.2.2.2. --- Serum Supplements --- p.47 / Chapter 3.2.2.3. --- Anti-CD16 Magnetic Microbeads --- p.47 / Chapter 3.2.2.4. --- Fico´HёØ-Paque Plus Solution --- p.47 / Chapter 3.2.2.5. --- PercolĺёØ Solution --- p.48 / Chapter 3.2.2.6. --- Phosphate Buffered Saline (PBS) --- p.48 / Chapter 3.2.2.7. --- Water --- p.48 / Chapter 3.2.3. --- Dye for Cellular Staining --- p.48 / Chapter 3.2.3.1. --- HemacoloŕёØ for Microscopy --- p.48 / Chapter 3.2.3.2. --- Trypan Blue Dye --- p.49 / Chapter 3.2.4. --- Reagents for Flow Cytometry --- p.49 / Chapter 3.2.4.1. --- FACS Flow Sheath Fluid --- p.49 / Chapter 3.2.4.2. --- FACS Wash Medium --- p.49 / Chapter 3.2.4.3. --- Paraformaldehyde --- p.49 / Chapter 3.2.5. --- Special Equipments and Apparatus --- p.49 / Chapter 3.2.5.1. --- Magnetic Cell Sorting System (MACS) --- p.49 / Chapter 3.3. --- Human Subjects --- p.51 / Chapter 3.3.1. --- Inclusion Criteria --- p.51 / Chapter 3.3.2. --- Exclusion Crtieria --- p.51 / Chapter 3.3.3. --- Medication --- p.52 / Chapter 3.3.4. --- Informed Consent and Patient Information --- p.52 / Chapter 3.4. --- Animals --- p.53 / Chapter 3.4.1. --- Maintenance --- p.53 / Chapter 3.4.2. --- Survival Experiment Using Erhlich Ascites Tumor Bearing ICR Mice --- p.53 / Chapter 3.4.3. --- Experiments of Immunomodulatory activity in Sarcoma 180 Bearing BALB/c Mice --- p.54 / Chapter 3.5. --- Methodology --- p.55 / Chapter 3.5.1. --- Preparation of the Traditional Chinese Medicine --- p.55 / Chapter 3.5.1.1. --- Hot Water Extraction of Water Soluble Fraction of Natural Cordyceps sinensis --- p.55 / Chapter 3.5.1.2. --- Hot Water Extraction of Water Soluble Fraction of HERBSnSENSEŚёØ Corydceps and the Wheeze-relief Formula for In vitro Experiments --- p.55 / Chapter 3.5.1.3. --- HERBSnSENSEŚёØ Corydceps for the In Vivo Experiments --- p.56 / Chapter 3.5.1.4. --- Extraction Efficiency of the Hot Water Extracts --- p.56 / Chapter 3.5.2. --- Limulus Ameobocyte Lysate Test --- p.56 / Chapter 3.5.3. --- Cell Preparation --- p.57 / Chapter 3.5.3.1. --- "Isolation of Human Peripheral Blood Mononuclear Cells, Lymphocytes and Monocytes" --- p.57 / Chapter 3.5.3.2. --- Isolation of Eosinophils --- p.58 / Chapter 3.5.3.3. --- Isolation of Spleen Cells from BALB/c Mice --- p.58 / Chapter 3.5.3.4. --- "Murine Ehrlich Ascites Tumor (EAT), PU5-18, and Sarcoma 180 (SC-180) Cell Lines" --- p.59 / Chapter 3.5.3.5. --- Human Eosinophilic Leukemic Cell Line (EoL-1) --- p.59 / Chapter 3.5.3.6. --- Human Hepatocarcinoma Hep-3B Cell Line --- p.59 / Chapter 3.5.3.7. --- Human Leukemic Cell Line (HL-60) --- p.59 / Chapter 3.5.3.8. --- Human Mast Cell Line (HMC-1) --- p.60 / Chapter 3.5.4. --- Collection of Mouse Serum and Human Plasma --- p.60 / Chapter 3.5.5. --- Collection of Culture Supernatant --- p.60 / Chapter 3.5.6. --- The Trypan Blue Exclusion Assay --- p.61 / Chapter 3.5.7. --- Colorimetric 5-bromo-2'-deoxyuridine (BrdU) Cell Proliferation Enzyme Linked Immunosorbent Assay (ELISA) --- p.61 / Chapter 3.5.8. --- Immunophenotyping --- p.62 / Chapter 3.5.9. --- The Cytometric Bead Array (CBA) Kits --- p.62 / Chapter 3.5.10. --- Intracellular Florescence Staining for Reactive Oxygen Species --- p.63 / Chapter 3.5.11. --- The Intracellular Zymosan Florescence Assay --- p.64 / Chapter 3.5.12. --- Total Cellular RNA Extraction --- p.64 / Chapter 3.5.13. --- Gel Electrophoresis of RNA Integrity --- p.65 / Chapter 3.5.14. --- cDNA Expression Array --- p.65 / Chapter 3.5.15. --- Cell Staining Using Cytospin --- p.66 / Chapter 3.5.16. --- Annexin V-FITC/Propidium Iodide Apoptosis Detection --- p.66 / Chapter 3.5.17. --- Weighing the Spleen and Tumor --- p.67 / Chapter 3.5.18. --- Preparing Samples for the Eosinophilic Cationic Protein Fluoroenzymeimmunoassay --- p.67 / Chapter 3.5.19. --- Statistical Analysis --- p.67 / Chapter Part III - --- Results: Pre-functional Assays / Chapter Chapter 4: --- "Extraction, Endotoxin Measurement, In vitro Cytotoxicity Testing, and the Selection of Optimal Concentration" / Chapter 4.1. --- Extraction efficiency --- p.68 / Chapter 4.1.1. --- Introduction --- p.68 / Chapter 4.1.2. --- Results --- p.68 / Chapter 4.2. --- Endotoxin Level --- p.69 / Chapter 4.2.1. --- Introduction --- p.69 / Chapter 4.2.2. --- Results and Interpretation --- p.69 / Chapter 4.3. --- Cytotoxicity --- p.70 / Chapter 4.3.1. --- Introduction --- p.70 / Chapter 4.3.2. --- Results and Interpretation --- p.71 / Chapter 4.3.2.1. --- Peripheral Blood Mononuclear Cells (PBMC) --- p.71 / Chapter 4.3.2.2. --- Eosinophils --- p.72 / Chapter 4.4. --- The Optimal Concentration (OC) --- p.76 / Chapter 4.4.1. --- Introduction --- p.76 / Chapter 4.4.2. --- Results and Interpretation --- p.76 / Chapter Part IV- --- Results: Immunomodulatory Activities of Cordyceps sinensis as a Single Herb / Chapter Chapter 5: --- Mitogenic Activity --- p.80 / Chapter 5.1. --- Introduction --- p.80 / Chapter 5.2. --- Results --- p.80 / Chapter 5.3. --- Discussion --- p.81 / Chapter Chapter 6: --- Cytokines and Cytokine Receptors --- p.84 / Chapter 6.1. --- Introduction --- p.84 / Chapter 6.2. --- Results --- p.84 / Chapter 6.2.1. --- Effects of Natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on the Induction of Cytokines from Lymphocytes --- p.84 / Chapter 6.2.1.1. --- TNFa --- p.84 / Chapter 6.2.1.2. --- IL-6 --- p.85 / Chapter 6.2.1.3. --- IL-10 --- p.85 / Chapter 6.2.2. --- Effects of Natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on the Induction of Cytokines from Monocytes --- p.92 / Chapter 6.2.2.1. --- IL-1β --- p.92 / Chapter 6.2.2.2. --- IL-6 --- p.92 / Chapter 6.2.2.3. --- IL-10 --- p.97 / Chapter 6.2.2.4. --- TNFα --- p.97 / Chapter 6.2.3. --- Effects of Natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on the Expression of Cytokine Receptor --- p.102 / Chapter 6.2.4. --- Effects of Natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on the Gene Expression of Cytokines and Cytokine Receptors in Peripheral Blood Mononuclear Cells --- p.105 / Chapter 6.3. --- Discussion --- p.112 / Chapter Chapter 7: --- Macrophage Functions: Phagocytosis and Release of Reactive Oxygen Species (ROS) --- p.116 / Chapter 7.1 --- Introduction --- p.116 / Chapter 7.2. --- Results --- p.117 / Chapter 7.2.1. --- Phagocytosis --- p.117 / Chapter 7.2.2. --- Release of Reactive Oxygen Species (ROS) --- p.117 / Chapter 7.3. --- Discussion --- p.124 / Chapter Chapter 8: --- Apoptosis of Selected Cancer Cell Lines --- p.126 / Chapter 8.1. --- Introduction --- p.126 / Chapter 8.2. --- Results --- p.127 / Chapter 8.2.1. --- Differential Cytotoxic Effects of natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on Various Cancer Cell Lines In vitro --- p.127 / Chapter 8.2.2. --- Differential Anti-Proliferative Effects of natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on Various Cancer Cell Lines In vitro --- p.129 / Chapter 8.2.3. --- Differential Apoptotic Effects of Natural Cordyceps sinensis and HERBSnSENSEŚёØ Cordyceps on Various Cancer Cell Lines In vitro --- p.131 / Chapter 8.2.3.1. --- Peripheral Blood Mononuclear Cells --- p.131 / Chapter 8.2.3.2. --- Hepatocarcinoma Hep-3B --- p.131 / Chapter 8.2.3.3. --- Human Eosinophilic Leukemic Cell Line --- p.134 / Chapter 8.2.3.4. --- Human Mast Cell Line --- p.134 / Chapter 8.2.3.5. --- Human Leukemic Cell Line (HL-60) --- p.138 / Chapter 8.2.3.6. --- Murine Macrophages/Monocytes Cell Line PU5-18 --- p.138 / Chapter 8.2.3.7. --- Murine Erhlich Ascites Tumor (EAT) --- p.142 / Chapter 8.2.3.8. --- Murine Sarcoma 180 (SC-180) --- p.142 / Chapter 8.3. --- Discussion --- p.145 / Chapter Part V- --- Results: Immunomodulatory Activities of Cordyceps sinensis in Concoction / Chapter Chapter 9: --- The In vivo Animal Model --- p.147 / Chapter 9.1. --- introduction --- p.147 / Chapter 9.2. --- Results --- p.148 / Chapter 9.2.1. --- The ICR Mice Model --- p.148 / Chapter 9.2.1.1. --- In vivo Effects of Natural C. sinensis and HERBSnSENSEŚёØ Cordyceps on the Ascitic Fluid Production of ICR Mice --- p.148 / Chapter 9.2.1.2. --- Effects of Natural C. sinensis and HERBSnSENSEŚёØ Cordyceps on the Survival of Tumor-bearing ICR Mice --- p.149 / Chapter 9.3. --- The BALB/c Mice Model --- p.153 / Chapter 9.3.1. --- In vivo Effects of HERBSnSENSEŚёØ Cordyceps on Spleen and Tumor Weight --- p.153 / Chapter 9.3.2. --- Effects of HERBSnSENSEŚёØ Cordyceps on the Mitogenic Activities of Spleen Cells --- p.154 / Chapter 9.3.3. --- "In vivo Effects of HERBSnSENSEŚёØ Cordyceps on the Cell Surface Expression of CD3, CD4, and CD8" --- p.157 / Chapter 9.3.4. --- Effects of HERBSnSENSEŚёØ Cordyceps on the Cytokine Release from Cultured Spleen Cells --- p.161 / Chapter 9.3.4.1. --- TNFα --- p.161 / Chapter 9.3.4.2. --- IFNγ --- p.163 / Chapter 9.3.4.3. --- IL-2 --- p.163 / Chapter 9.3.4.4. --- IL-4 --- p.163 / Chapter 9.3.4.5. --- IL-6 --- p.167 / Chapter 9.3.4.6. --- IL-10 --- p.167 / Chapter 9.3.4.7. --- IL-12p70 --- p.167 / Chapter 9.3.4.8. --- Monocyte Chemoattractant Protein(MCP)-1 --- p.167 / Chapter 9.3.5. --- In vivo Effects of HERBSnSENSEŚёØ Cordyceps on the Cytokine Synthesis --- p.172 / Chapter 9.4. --- Discussion --- p.174 / Chapter Chapter 10: --- In vitro Studies on Eosinophils and Peripheral Blood Mononuclear Cells --- p.178 / Chapter 10.1. --- Introduction --- p.178 / Chapter 10.2. --- Results --- p.180 / Chapter 10.2.1. --- In vitro Effects of Wheeze-Relief Formula on the Survival of IL-5 Enhanced Eosinophils --- p.180 / Chapter 10.2.2. --- In vitro Effects of Wheeze-Relief Formula on the Degranulation of Eosinophils --- p.180 / Chapter 10.2.3. --- In vitro Effects of Wheeze-Relief Formula on the Surface Expression of Adhesion Molecules and Chemokine Receptors on Eosinophils --- p.183 / Chapter 10.2.4. --- In vitro Effects of Wheeze-Relief Formula on the Surface Expression of Adhesion Molecules on Eosinophils --- p.183 / Chapter 10.2.5. --- In vitro Effects of Wheeze-Relief Formula on the Cytokine Release from Peripheral Blood Mononuclear Cells --- p.187 / Chapter 10.2.6. --- In vitro Effects of Wheeze-Relief Formula on the Gene Expression Profile of Cytokines and Cytokine Receptors of Peripheral Blood Mononuclear Cells --- p.187 / Chapter 10.3. --- Discussion --- p.196 / Chapter Chapter 11: --- The Clinical Trial: Analysis of Serological Markers --- p.200 / Chapter 11.1. --- Introduction --- p.200 / Chapter 11.2. --- Results --- p.202 / Chapter 11.2.1. --- Demographic Data and Drop-out Cases --- p.202 / Chapter 11.2.2. --- Lung Function Test --- p.202 / Chapter 11.2.3. --- Steroid Dosage --- p.202 / Chapter 11.2.4. --- Serological Markers --- p.205 / Chapter 11.3. --- Discussion --- p.215 / Chapter Part VI - --- Conclusion / Chapter Chapter 12: --- Concluding Remarks and Future Perspectives --- p.217 / Chapter Part VII- --- Appendix / Parent Information Sheet --- p.222 / 家長資訊 --- p.223 / Consent Form --- p.224 / Licence to Conduct Animal Experiments --- p.225 / Bibliography --- p.227
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324857 |
| Date | January 2004 |
| Contributors | Lee, Ka Wai Sharon., Chinese University of Hong Kong Graduate School. Division of Chemical Pathology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xxvii, 260 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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