Li Tin Wai Olive. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 191-217). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.II / ABSTRACT --- p.III / 中文摘要 --- p.IX / PUBLICATIONS --- p.XIV / TABLE OF CONTENTS --- p.XV / LIST OF ABBREVIATIONS --- p.XXI / LIST OF FIGURES AND TABLES --- p.XXIII / Chapter CHAPTER 1. --- INTRODUCTION --- p.1 / Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.7 / Chapter 2.1. --- Cardiovascular disease --- p.7 / Chapter 2.1.1. --- Introduction --- p.7 / Chapter 2.1.2. --- Atherosclerosis --- p.7 / Chapter 2.1.3. --- Cellular and molecular deregulation in early atherosclerosis --- p.10 / Chapter 2.1.3.1. --- Introduction --- p.10 / Chapter 2.1.3.2. --- Endothelial dysfunction --- p.11 / Chapter 2.1.3.3. --- Nitric oxide --- p.12 / Chapter 2.1.3.4. --- Adhesion molecules and the early events of atherogenesis --- p.13 / Chapter 2.1.3.4.1. --- Introduction --- p.13 / Chapter 2.1.3.4.2. --- Intracellular adhesion molecule-1 --- p.15 / Chapter 2.1.3.4.3. --- Nuclear factor kappa B --- p.18 / Chapter 2.1.3.5. --- Summary --- p.19 / Chapter 2.2. --- Nitric oxide in molecular vascular biology --- p.20 / Chapter 2.2.1. --- Introduction --- p.20 / Chapter 2.2.2. --- Nitric oxide synthase --- p.21 / Chapter 2.2.2.1. --- Introduction --- p.21 / Chapter 2.2.2.2. --- Endothelial nitric oxide synthase --- p.24 / Chapter 2.2.2.3. --- Inducible nitric oxide synthase --- p.25 / Chapter 2.2.2.4. --- Nitric oxide concentration dependent effector pathways --- p.26 / Chapter 2.2.3. --- Nitric oxide and its regulation in vascular events --- p.28 / Chapter 2.2.3.1. --- Introduction --- p.28 / Chapter 2.2.3.2. --- Regulation of vascular tone --- p.30 / Chapter 2.2.3.3. --- "Regulation of platelet adhesion, activation and aggregation" --- p.32 / Chapter 2.2.3.4. --- Regulation of endothelial adhesiveness and leukocyte adhesion - Anti-adhesive effect of nitric oxide --- p.32 / Chapter 2.2.3.5. --- "Regulation of vascular smooth muscle growth, migration and proliferation" --- p.33 / Chapter 2.2.3.6. --- Antioxidative effect of nitric oxide --- p.34 / Chapter 2.2.3.7. --- Regulation of endothelial apoptosis --- p.35 / Chapter 2.2.3.8. --- Nitric oxide and its relationship with other risk factors --- p.36 / Chapter 2.3. --- "Menopause, cardiovascular diseases and Traditional Chinese Medicine" --- p.37 / Chapter 2.3.1. --- Traditional Chinese Medicine and menopause --- p.37 / Chapter 2.3.2. --- Dang Gui Buxue Tang --- p.38 / Chapter 2.3.3. --- Danggui --- p.39 / Chapter 2.3.3.1. --- Botanic origins --- p.39 / Chapter 2.3.3.2. --- Usage --- p.39 / Chapter 2.3.4. --- Huangqi --- p.40 / Chapter 2.3.4.1. --- Botanic origins --- p.40 / Chapter 2.3.4.2. --- Usage --- p.40 / Chapter 2.3.5. --- Modern scientific research --- p.41 / Chapter 2.3.5.1. --- General cardioprotective role --- p.41 / Chapter 2.3.5.2. --- Vascular tone modulation --- p.42 / Chapter 2.3.5.3. --- Haemostasis --- p.42 / Chapter 2.3.5.4. --- Endothelial cell --- p.43 / Chapter 2.3.5.4.1. --- Nitric oxide pathway --- p.43 / Chapter 2.3.5.4.1.1. --- Direct alteration of nitric oxide secretion --- p.43 / Chapter 2.3.5.4.1.2. --- Alteration of Nitric oxide synthase expression or activity --- p.43 / Chapter 2.3.5.4.2. --- Alteration of adhesion molecule expression --- p.44 / Chapter 2.3.5.4.3. --- Alteration of adhesion molecule expression as an effect of nitric oxide secretion --- p.45 / Chapter 2.3.5.5. --- Antioxidant effect --- p.45 / Chapter 2.3.5.6. --- Estrogenicity of DBT --- p.46 / Chapter 2.4. --- Research plan --- p.47 / Chapter 2.4.1. --- Formulation of research hypotheses --- p.47 / Chapter 2.4.1.1. --- Hypotheses --- p.50 / Chapter 2.4.2. --- Plan of study --- p.50 / Chapter 2.4.2.1. --- Dang Gui Buxue Tang extraction and standardization of content --- p.50 / Chapter 2.4.2.2. --- Cell model development --- p.52 / Chapter 2.4.2.3. --- Experimental studies --- p.54 / Chapter 2.4.2.4. --- eNOS activity determination - the nitric oxide metabolite assay --- p.56 / Chapter 2.4.2.5. --- Endotoxin contamination in DBT --- p.57 / Chapter 2.4.3. --- Sample size and statistical analysis --- p.59 / Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.62 / Chapter 3.1. --- Dang Gui Buxue Tang extraction and content standardization --- p.62 / Chapter 3.1.1. --- Plant materials --- p.62 / Chapter 3.1.2. --- DBT authentication --- p.62 / Chapter 3.1.3. --- DBT processing prior to extraction --- p.63 / Chapter 3.1.4. --- DBT extraction --- p.63 / Chapter 3.1.5. --- Quantitative standardization of DBT markers by High Pressure Liquid Chromatography --- p.66 / Chapter 3.1.5.1. --- DBT markers: standard preparation --- p.66 / Chapter 3.1.5.2. --- Sample preparation --- p.67 / Chapter 3.1.5.3. --- Quantitative analysis of DBT constituents by HPLC --- p.67 / Chapter 3.1.6. --- DBT polysaccharide standardization --- p.68 / Chapter 3.1.6.1. --- Glucose standard preparation --- p.68 / Chapter 3.1.6.2. --- Sample preparation --- p.68 / Chapter 3.1.6.3. --- Quantitative determination of polysaccharide by Phenol-Sulfuric acid colorimetric assay --- p.68 / Chapter 3.1.7. --- DBT endotoxin contamination determination --- p.69 / Chapter 3.1.7.1. --- "Positive, negative and inhibition controls" --- p.69 / Chapter 3.1.7.2. --- Qualitative determination of sample endotoxin --- p.70 / Chapter 3.2. --- Cell culture --- p.70 / Chapter 3.2.1. --- Characterization of cultured cells --- p.72 / Chapter 3.2.2. --- Passage --- p.73 / Chapter 3.3. --- DBT treatment --- p.73 / Chapter 3.3.1. --- Solvent system of DBT treatment --- p.73 / Chapter 3.3.2. --- Dosage and duration of DBT treatment --- p.74 / Chapter 3.3.3. --- Positive and negative controls --- p.74 / Chapter 3.4. --- MTT-based cytotoxicity assay --- p.75 / Chapter 3.5. --- Reverse transcriptase- polymerase chain reaction --- p.76 / Chapter 3.5.1. --- Sample preparation --- p.76 / Chapter 3.5.1.1. --- Total RNA isolation --- p.76 / Chapter 3.5.1.2. --- DNase treatment --- p.77 / Chapter 3.5.1.3. --- RNAethanol precipitation --- p.78 / Chapter 3.5.1.4. --- Complementary DNA synthesis --- p.78 / Chapter 3.5.2. --- Polymerase chain reaction --- p.79 / Chapter 3.5.2.1. --- Polymerase chain reaction conditions --- p.79 / Chapter 3.5.2.2. --- Primers --- p.79 / Chapter 3.5.3. --- Visualization of the PCR products --- p.81 / Chapter 3.5.3.1. --- Gel electrophoresis --- p.81 / Chapter 3.5.3.2. --- Gel Doc software --- p.82 / Chapter 3.5.3.3. --- Densitometry --- p.82 / Chapter 3.5.4. --- Real time RT-PCR --- p.82 / Chapter 3.6. --- Quantitative Immunocytochemical studies --- p.84 / Chapter 3.6.1. --- Coverslip preparation --- p.84 / Chapter 3.6.2. --- Sample preparation --- p.84 / Chapter 3.6.3. --- Immunocytochemical staining preparation --- p.85 / Chapter 3.6.3.1. --- "Immunocytochemical staining for vWF, α-actin, iNOS, ICAM-1, NF-kB using DAKO catalyzed signal amplification (CSA) system (anti-mouse)" --- p.86 / Chapter 3.6.3.2. --- Immunocytochemical staining for eNOS using Santa Cruz immunoCruz staining system (anti-goat) --- p.87 / Chapter 3.6.4. --- Counterstaining and mounting --- p.88 / Chapter 3.6.5. --- Result interpretation --- p.89 / Chapter 3.6.5.1. --- Microscopy and digital image capture --- p.89 / Chapter 3.6.5.2. --- Determination of Image (file) Energy --- p.89 / Chapter 3.7. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.90 / Chapter 3.7.1. --- Sample preparation --- p.90 / Chapter 3.7.2. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.91 / Chapter CHAPTER 4. --- RESULTS --- p.93 / Chapter 4.1. --- Dang Gui Buxue Tang extraction and standardization of content --- p.93 / Chapter 4.1.1. --- DBT extraction - general data --- p.93 / Chapter 4.1.2. --- DBT polysaccharide standardization --- p.97 / Chapter 4.1.3. --- DBT marker standardization --- p.101 / Chapter 4.1.4. --- DBT endotoxin contamination determination --- p.104 / Chapter 4.2. --- Cell model development --- p.108 / Chapter 4.2.1. --- Endothelial morphology --- p.108 / Chapter 4.2.2. --- Immunocytochemistiy --- p.108 / Chapter 4.2.3. --- MTT cytotoxicity assay --- p.110 / Chapter 4.3. --- Study 1 --- p.112 / Chapter 4.3.1. --- Immunocytochemistry (Hypothesis 1) --- p.112 / Chapter 4.3.2. --- RT-PCR (Hypothesis 1) --- p.117 / Chapter 4.3.3. --- Real time RT-PCR (Hypothesis 1) --- p.121 / Chapter 4.3.4. --- Immunocytochemistry (Hypothesis 2) --- p.125 / Chapter 4.3.5. --- RT-PCR (Hypothesis 2) --- p.128 / Chapter 4.3.6. --- Total Nitrite/Nitrate quantitative colorimetric assay --- p.131 / Chapter 4.4. --- Study 2 --- p.133 / Chapter 4.4.1. --- Immunocytochemistry --- p.133 / Chapter 4.5. --- Study 3 --- p.138 / Chapter 4.5.1. --- Immunocytochemistry --- p.138 / Chapter 4.5.2. --- RT-PCR --- p.141 / Chapter 4.5.3. --- Real time RT-PCR --- p.145 / Chapter 4.6. --- Endotoxin contamination in DBT --- p.149 / Chapter 4.6.1. --- Effects of endotoxin on eNOS --- p.149 / Chapter 4.6.2. --- Immunocytochemistry on immunostained endothelial cells --- p.151 / Chapter 4.6.3. --- Effects of endotoxin on iNOS --- p.153 / Chapter 4.6.4. --- Effect of endotoxin on NF-kB --- p.157 / Chapter 4.6.5. --- Effects of endotoxin on ICAM-1 --- p.160 / Chapter CHAPTER 5. --- DISCUSSION --- p.165 / Chapter 5.1. --- DBT extraction and standardization of content --- p.165 / Chapter 5.1.1. --- Optimal DBT extraction conditions --- p.165 / Chapter 5.1.2. --- Evidence to support formulae usage --- p.166 / Chapter 5.1.3. --- Limitation of the methodology used --- p.166 / Chapter 5.2. --- Cell model development --- p.167 / Chapter 5.2.1. --- Choice of DBT concentration range in the study --- p.167 / Chapter 5.2.1.1. --- Choice of concentration range in consideration of endotoxin contamination --- p.167 / Chapter 5.2.1.2. --- Choice of concentration range in consideration of DBT's cytotoxicity effects --- p.168 / Chapter 5.2.1.3. --- Choice of concentration range in consideration of prevous studies --- p.168 / Chapter 5.3. --- Study 1 --- p.169 / Chapter 5.3.1. --- Action of DBT on eNOS expression --- p.169 / Chapter 5.3.2. --- Action of DBT on iNOS expression --- p.170 / Chapter 5.3.3. --- Action of DBT on Nitric oxide metabolite assay --- p.171 / Chapter 5.3.3.1. --- Result interpretation with rejected hypothesis 2 --- p.171 / Chapter 5.3.3.2. --- Assay limitations and improvements --- p.171 / Chapter 5.4. --- Study 2 --- p.172 / Chapter 5.4.1. --- Action of DBT on NF-kB expression --- p.172 / Chapter 5.4.2. --- Assay limitations and improvements --- p.173 / Chapter 5.5. --- Study 3 --- p.174 / Chapter 5.5.1. --- Action of DBT on ICAM-1 expression --- p.174 / Chapter 5.6. --- Endotoxin contamination in DBT --- p.175 / Chapter 5.6.1. --- Action of endotoxin contamination in DBT on various markers --- p.175 / Chapter 5.6:2. --- Experimental limitation --- p.176 / Chapter 5.6.3. --- Endotoxin removal --- p.177 / Chapter 5.6.3.1. --- Introduction --- p.177 / Chapter 5.6.3.2. --- Endotoxin removal methodologies suitable for herbal use --- p.179 / Chapter 5.7. --- Action of DBT on angiogenesis stimulation --- p.181 / Chapter 5.7.1. --- Evidence for DBT's proangiogenic effects from various studies --- p.181 / Chapter 5.7.2. --- Influence of endotoxin contamination on angiogenesis stimulation --- p.182 / Chapter 5.7.3. --- Assay limitations and future developments --- p.183 / Chapter CHAPTER 6. --- GENERAL DISCUSSION AND SUMMARY --- p.186 / Chapter CHAPTER 7. --- REFERENCES --- p.191

Thesis (M. Arch.)--University of Hong Kong, 2002. / Includes special report study entitled: Materia medica. Includes bibliographical references.

Thesis (M. Sc.)--University of Hong Kong, 2000. / Includes bibliographical references (leaves 83-87).

Thesis (M.Arch.)--University of Hong Kong, 2002. / Includes special report study entitled : Materia medica. Includes bibliographical references. Also available in print.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

published_or_final_version / Psychiatry / Master / Master of Medical Sciences

by Law Lap Kay. / Bibliography: leaves 96-106 / Thesis (M.Phil.) -- Chinese University of Hong Kong, 1983

by Chan Wood-yee. / Bibliography: leaves 129-146 / Thesis (M.Ph.)--Chinese University of Hong Kong, 1984

Chemical analyses, biological assays, and animal experiments provide important fundamental information of Traditional Chinese Medicine. In modern biomedical research, they should also be necessary prerequisites for clinical trials. A Chinese herbal formula, no matter how effective and safe in the pre-clinical studies, cannot be assumed clinically valid. No marketing approval should be granted without clinical studies. Thus, clinical trial plays a decisive role in the research and development of Traditional Chinese Medicine. Currently the highest level of efficacy evidence is obtained from randomized controlled clinical trial. Thousands of years of traditional use can provide us with valuable guidelines to the selection, preparation and application of herbal formulations. To be accepted as viable alternatives to western medicine, the rigorous methods of scientific and clinical validations must be applied. / In conclusion, the methodology of Chinese medicine research needs to improve. One practical way is to apply the efficacy-driven approach through the following steps: i) getting a simple herbal formula to try solving a difficult clinical problem and start an evidence-based clinical trial using methodology acceptable to current standard clinical trials i.e. randomized, placebo-controlled; ii) parallel laboratory experiments to understand the mode of action should be done; and; iii) making sure that the quality of herbs and their extracts are of the best standard. A promising item shown in the clinical trial and laboratory experiments would deserve more pharmacological investigations before considering marketing. (Abstract shortened by UMI.) / The distinctiveness of Chinese medicine is manifested in the diversity and the complexity of its components, the instability of its quantity, the fuzziness of its action mechanism, and the uncontrollability of its producing process. Traditional Chinese herbal formulae are usually formed by more than one plants, animal or mineral items. The composition is extremely complex. The efficacy thus can hardly be guaranteed. The methods of harvesting, drying, storage, transportation, and processing of plant material influence the efficacy and safety. The consistent efficacy, therefore, can hardly be guaranteed. For thousands of years, it has been observed by clinical practice that Traditional Chinese Medicine (TCM) has a rich scientific connotation and has developed a unique healthcare system. However, variable sources of raw materials, unknown active ingredients, difficulties in quality control, lack of safety evaluation, unclear mechanism of action, etc., all these factors constitute major challenges in modernization of TCM. / The effect of Traditional Chinese Medicine (TCM) may be characterized by its chemical compounds, which are also the active ingredients. The reproducibility and the stability of the active ingredients are the foundation to ensure the efficacy of TCM. The safety and efficacy of TCM is evaluated through its pharmacological effects and in clinical studies. Many Chinese herbal medicines have a long history of traditional use. However, most of them are of unproven efficacy by today's standard. Well-designed randomized controlled trials and comprehensive pre-clinical studies are not known. Although the lack of qualified evidence does not mean that Chinese herbal medicines lack efficacy or are unsafe, properly designed experimental and clinical investigations should still be done today. There is a need to scientifically prove and clinically validate its safety and efficacy through chemical standardization, biological assays, and clinical trials. / The general perception that herbal drugs are very safe and free from side effects is not true. Little do they know in the modern conditions, that the current herbs used for preparation of traditional Chinese medicine have been very different as compared with the ancient herbs in planting, extracting, producing, storage, application, dose level and the duration of clinical use. The active ingredients of herbal formula are higher after extracting with modern scientific methods; and the toxicity may also be correspondingly higher. The potential side effects after long-term use should not be overlooked. / The purpose of the research is to develop a practical methodology to obtain convincing evidences in quality, safety and efficacy of Chinese Medicine. When we use evidence-based research methodology to prove the quality, safety and efficacy, we are facing many challenges. / The quality control of Chinese herbal medicine is a systematic procedure. The initial critical step is to standardize the starting raw herbs, for knowing the exact species and subspecies, the ideal growing location, environmental conditions, harvesting methods, and storage conditions etc., i.e., following Good Agriculture Practice (GAP) to ensure the quality of the raw materials. The second step is to standardize the processing methods of the raw herbs. The third step is to standardize the preparation procedure of the final products according to the requirements of Good Manufacture Practice (GMP) guideline. The fourth step is to qualitatively and/or quantitatively evaluate the quality of the TCM medication based on one or more selected chemical markers. / by Cheng, King Fai. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Text in English with some Chinese characters; abstract and appendixes 3-5 also in Chinese.

Thesis (M. D. S.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.