Lee Kwing Chin, Kenneth. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 298-357). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Diabetes mellitus has been associated with both clinical and experimental cardiac dysfunction. Diabetic cardiomyopathy which is characterized by depressed cardiac contractility is accompanied by a variety of biochemical changes in Ca⁺⁺ metabolism. This cardiomyopathy may occur in the presence of normal coronary arteries and normal blood pressure. However, some studies have shown that hypertension is more prevalent among diabetics and can aggravate the cardiovascular abnormalities associated with diabetes. To understand the mechanisms of diabetic cardiomyopathy and consequences of combined hypertension and diabetes, experiments were designed to measure cardiac tissue responses to various inotropic agents in experimental diabetes. Six weeks following streptozotocin (STZ) administration, Wistar, spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats exhibited the 'classical signs' of diabetes which included: hyperglycemia, hypoinsulinemia, hyperlipidemia (except in WKY), and hypothyroidism. Decreased basal atrial rate and increased basal developed force (BDF) suggest a depressed SA node function and an alteration of Ca⁺⁺ utilization by diabetic ventricles. Decreased post quiescent potentiation (PQP) values (except in WKY) in ventricular tissues suggest a diminished amount of releasable Ca⁺⁺ from sarcoplasmic reticulum (SR). Decreased post stimulation potentiation (PSP) values in SHR papillary muscles (PM) are probably suggestive of a depressed sarcolemmal Na⁺-Ca⁺⁺ exchange function in this tissue. Diabetic rats show subsensitivity to β-adrenergic stimulation in ventricular tissues, supersensitivity and hyperresponsiveness to Ca⁺⁺ and α-adrenergic stimulation (except in WKY) in ventricular tissues and left atria (LA) and supersensitivity to BAY K 8644 in SHR LA and hyperresponsiveness to verapamil in ventricular strips. These alterations may be attributed to a change in receptor number and/or a post receptor alteration. Ryanodine decreased the PQP of Wistar and SHR PM and SHR LA in both controls and diabetics. It especially abolished PQP in SHR diabetic tissues, but had no effect on WKY tissues, which may suggest a difference in the SR function in these tissues. SR with impaired Ca⁺⁺ uptake may contribute to these phenomena in diabetic rats. Ryanodine also diminished (PQP + BDF) of SHR LA and (PQP/BDF) of Wistar and SHR PM, ˙but had no effects on control and other diabetic tissues. It appears that ryanodine has some influence on the Na⁺-Ca⁺⁺ exchange generated by sarcolemma (SL) of certain diabetic tissues. Further experiments are required to clarify this. SHR diabetic rats had greater changes in most of the measurements such as hyperlipidemia, depressed PQP and PSP values, and altered drug responses. This model exhibited very high mortality as compared to Wistar and WKY diabetic rats. As has been shown previously, the combination of hypertension and diabetes exerts a synergistic effect on the cardiac dysfunction in this model, and that altered lipid metabolism, SL and SR function are all involved in the development of cardiomyopathy. WKY diabetic rats, on the other hand, exhibited no significant changes in blood lipids, or in response to phenylephrine or to Ca⁺⁺ (LA) stimulation. Lack of change in these factors may explain the relatively normal cardiac function of this model as measured previously. / Pharmaceutical Sciences, Faculty of / Graduate

by Lam Fung Chun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 187-202). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.xiii / List of Tables --- p.xvii / List of Figures --- p.xviii / Publication --- p.xx / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Epidemiology of Diabetes Mellitus --- p.1 / Chapter 1.2 --- Definition of Diabetes Mellitus --- p.1 / Chapter 1.3 --- Glucose Homeostasis and Diabetes Mellitus --- p.2 / Chapter 1.4 --- Classification of Diabetes Mellitus --- p.6 / Chapter 1.4.1 --- Type 1 Diabetes Mellitus --- p.6 / Chapter 1.4.2 --- Type 2 Diabetes Mellitus --- p.7 / Chapter 1.4.3 --- Gestational Diabetes Mellitus --- p.8 / Chapter 1.4.4 --- Other specific types --- p.8 / Chapter 1.5 --- Diagnostic Criteria of Diabetes Mellitus --- p.9 / Chapter 1.6 --- Complications of Diabetes Mellitus --- p.11 / Chapter 1.7 --- Pharmacological Treatment of Diabetes --- p.12 / Chapter 1.7.1 --- Treatment for type 1 diabetes mellitus --- p.12 / Chapter 1.7.2 --- Treatment for Type 2 diabetes mellitus --- p.13 / Chapter 1.7.2.1 --- Sulfonylureas --- p.14 / Chapter 1.7.1.2 --- Meglitinides --- p.15 / Chapter 1.7.1.3 --- Biguanides --- p.15 / Chapter 1.7.1.4 --- Thazolidinediones --- p.16 / Chapter 1.7.1.5 --- α-Glucosidase inhibitor --- p.16 / Chapter 1.8 --- Diabetes and Traditional Chinese Medicine --- p.17 / Chapter 1.9 --- Objective of this project --- p.18 / Chapter Chapter 2 --- "Botanical, Preparation and Authentication of Traditional Chinese Herbs" --- p.22 / Chapter 2.1 --- Introduction --- p.22 / Chapter 2.2 --- Herbal Materials --- p.22 / Chapter 2.3 --- Authentication of Herbal Material --- p.30 / Chapter 2.4 --- Extraction Method --- p.32 / Chapter 2.4.1 --- Material and Methods --- p.32 / Chapter 2.4.2 --- Results --- p.32 / Chapter 2.4 --- Discussion --- p.32 / Chapter Chapter 3 --- In vitro Studies on Selected Traditional Chinese Herbs --- p.35 / Chapter 3.1. --- Introduction --- p.35 / Chapter 3.2 --- Hepatic Gluconeogenesis Studies --- p.36 / Chapter 3.2.1 --- Introduction --- p.36 / Chapter 3.2.2 --- Material and Methods --- p.41 / Chapter 3.2.2.1 --- Cell Culture of H4IIE --- p.41 / Chapter 3.2.2.2 --- Glucose Production Assay --- p.42 / Chapter 3.2.2.3 --- Bicinchoninic Acid (BCA) Protein Assay --- p.43 / Chapter 3.2.3 --- Results --- p.44 / Chapter 3.3 --- Intestinal Glucose Absorption Studies --- p.46 / Chapter 3.3.1 --- Introduction --- p.46 / Chapter 3.3.2 --- Material and Methods --- p.48 / Chapter 3.3.2.1 --- Preparation of BBMV --- p.48 / Chapter 3.3.2.1.1 --- Chemicals --- p.48 / Chapter 3.3.2.1.2 --- Method --- p.48 / Chapter 3.3.2.2 --- Preparation of Herbal Extracts --- p.50 / Chapter 3.3.2.3 --- BBMV Glucose Uptake Assay --- p.51 / Chapter 3.3.2.4 --- Bicinchoninic Acid (BCA) Protein Assay --- p.54 / Chapter 3.3.3 --- Results --- p.54 / Chapter 3.4 --- Fibroblast Glucose Uptake Studies --- p.57 / Chapter 3.4.1 --- Introduction --- p.57 / Chapter 3.4.2 --- Material and Methods --- p.58 / Chapter 3.4.2.1 --- Cell Culture of Hs68 --- p.58 / Chapter 3.4.2.2 --- 2-Deoxy-D-glucose Uptake Assay --- p.59 / Chapter 3.4.2.3 --- Bicinchoninic Acid (BCA) Protein Assay --- p.60 / Chapter 3.4.3 --- Results --- p.60 / Chapter 3.5 --- Adipocyte Glucose Uptake Studies --- p.63 / Chapter 3.5.1 --- Introduction --- p.63 / Chapter 3.5.2 --- Material and Methods --- p.65 / Chapter 3.5.2.1 --- Cell Culture of 3T3-L1 --- p.65 / Chapter 3.5.2.2 --- Differentiation of 3T3-L1 --- p.65 / Chapter 3.5.2.3 --- 2-Deoxy-D-glucose Uptake Assay --- p.66 / Chapter 3.5.2.4 --- Bicinchoninic Acid (BCA) Protein Assay --- p.68 / Chapter 3.5.3 --- Results --- p.69 / Chapter 3.6 --- Glucose Transporter Type 4 (GLUT4) Expression Studies --- p.71 / Chapter 3.6.1 --- Introduction --- p.71 / Chapter 3.6.2 --- Material and Methods --- p.48 / Chapter 3.6.2.1 --- Cell Culture of 3T3-L1 --- p.71 / Chapter 3.6.2.2 --- Differentiation of 3T3-L1 --- p.71 / Chapter 3.6.2.3 --- GLUT4 Expression Assay --- p.72 / Chapter 3.6.2.4 --- Preparation of RNA --- p.72 / Chapter 3.6.2.5 --- RT-PCR --- p.73 / Chapter 3.6.2.6 --- PCR Analysis on GLUT4 Expression --- p.74 / Chapter 3.6.2.7 --- Real-time PCR --- p.75 / Chapter 3.6.3 --- Results --- p.77 / Chapter 3.7 --- Discussion --- p.81 / Chapter 3.7.1 --- Discussion of Hepatic Gluconeogenesis Studies --- p.81 / Chapter 3.7.2 --- Discussion of Intestinal Glucose Absorption Studies --- p.82 / Chapter 3.7.3 --- Discussion of Fibroblast Glucose Uptake Studies --- p.83 / Chapter 3.7.4 --- Discussion of Adipocyte Glucose Uptake Studies --- p.84 / Chapter 3.7.5 --- Discussion of Glucose Transporter Type 4 (GLUT4) Expression Studies --- p.86 / Chapter 3.7.6 --- Conclusion --- p.87 / Chapter Chapter 4 --- Purification of Cortex Moutan --- p.90 / Chapter 4.1 --- Introduction --- p.90 / Chapter 4.1.1 --- Phytochemical Studies of Cortex Moutan --- p.90 / Chapter 4.2 --- Organic Extraction of Cortex Moutan --- p.93 / Chapter 4.2.1 --- Extraction Material and Methods --- p.93 / Chapter 4.2.2. --- Results --- p.93 / Chapter 4.3 --- BBMV Glucose Uptake Assay with Cortex Moutan Organic Extract (CM-C and CM-D) --- p.96 / Chapter 4.3.1 --- Material and Methods --- p.48 / Chapter 4.3.2 --- Results --- p.96 / Chapter 4.4 --- Fractionation of CM-C and CM-D --- p.98 / Chapter 4.4.1 --- Material and Methods --- p.98 / Chapter 4.4.1.1 --- Chemicals --- p.98 / Chapter 4.4.1.2 --- Methods --- p.98 / Chapter 4.4.2 --- Results --- p.100 / Chapter 4.5 --- BBMV Glucose Uptake Assay of CM-C and CM-D Sub-fractions --- p.105 / Chapter 4.5.1 --- Results --- p.105 / Chapter 4.6 --- Sulfonylation of CM-D1 --- p.107 / Chapter 4.6.1 --- Material and Methods --- p.107 / Chapter 4.6.1.1 --- Chemicals --- p.107 / Chapter 4.6.1.2 --- Methods --- p.107 / Chapter 4.6.2 --- Structure Elucidation of CM-D1s --- p.108 / Chapter 4.6.2.1 --- 1H-NMR Analysis --- p.108 / Chapter 4.6.3 --- BBMV Glucose Uptake Assay of CM-D1s --- p.108 / Chapter 4.6.4 --- Results --- p.108 / Chapter 4.7 --- "Structural Elucidation of CM-D3, CM-D4 and CM-D5" --- p.112 / Chapter 4.7.1 --- Material and Methods --- p.112 / Chapter 4.7.1.1 --- Mass Spectrometry --- p.112 / Chapter 4.7.1.2 --- 1H-NMR Analysis --- p.112 / Chapter 4.7.2 --- Results --- p.113 / Chapter 4.8 --- "BBMV Glucose Uptake Assay of Acetovallione, CM-D3，CM-D4 and CM-D5" --- p.116 / Chapter 4.8.1 --- Results --- p.116 / Chapter 4.9 --- Synthesis of CM-D3s --- p.118 / Chapter 4.9.1 --- Material and Methods --- p.118 / Chapter 4.9.1.1 --- Chemicals --- p.118 / Chapter 4.9.1.2 --- Methods --- p.118 / Chapter 4.9.2 --- Structure Elucidation of synthesized product --- p.119 / Chapter 4.9.3 --- Results --- p.119 / Chapter 4.10 --- Discussion --- p.121 / Chapter Chapter 5 --- In vivo Studies on Selected Herbs --- p.123 / Chapter 5.1 --- Introduction --- p.123 / Chapter 5.1.1 --- Diabetic Animal Models --- p.123 / Chapter 5.1.2 --- Neonatal Streptozotocin-induced Diabetic Rat Model --- p.125 / Chapter 5.2 --- Oral Glucose Tolerance Test (OGTT) --- p.126 / Chapter 5.2.1 --- Animal --- p.126 / Chapter 5.2.2 --- Rat Induction Material and Methods --- p.126 / Chapter 5.2.3 --- Testing Method for diabetic condition of rats --- p.127 / Chapter 5.3.4 --- Results --- p.128 / Chapter 5.3 --- Basal Glycaemia Test --- p.138 / Chapter 5.3.1 --- Animal --- p.138 / Chapter 5.3.2 --- Rat Induction Material and Methods --- p.138 / Chapter 5.3.3 --- Testing Method --- p.138 / Chapter 5.3.4 --- Results --- p.140 / Chapter 5.4 --- Discussion --- p.143 / Chapter Chapter 6 --- General Discussion --- p.147 / Chapter 6.1 --- Introduction --- p.147 / Chapter 6.2 --- Summary of Research Findings --- p.151 / Chapter 6.3 --- Result Interpretation --- p.152 / Chapter 6.3.1 --- Result Interpretation of In Vitro Studies --- p.152 / Chapter 6.3.2 --- Result Interpretation of Cortex Moutan Purification --- p.154 / Chapter 6.3.3 --- Result Interpretation of In Vivo Studies --- p.157 / Chapter 6.4 --- Limitations and Improvements --- p.161 / Chapter 6.5 --- Future Directions --- p.163 / Chapter 6.6 --- Conclusions --- p.169 / Appendices --- p.170 / References --- p.187

Basing on the traditional TCM interpretation, experience of recent research studies and our experimental findings, a few component herbs in Formulae 1 & 2 would be tentatively selected for a new formula. They were Radix Rehmanniae, Radix Astragali, Rhizoma Atractylodis Macrocephalae, Rhizoma Alismatis, Cortex Moutan and Rhizoma Smilacis Chinensis. Whether the new formula could give better efficacy would need to be tested in new clinical trials and experimental models. (Abstract shortened by UMI.) / Diabetes mellitus has long been a clinical problem for hundreds of years. More than 194 million people in the world now suffer from the disorder. About 15% of all diabetic patients would develop unhealing foot ulcers which compile significant proportion of nontraumatic lower-extremity amputations. Basing on the clinical experience of Prof. Xi Jiu Yi in Shanghai, literature review and an innovative interpretation of traditional Chinese medicine, two formulae (F1 & F2) derived from a well known herbal formula: the "Pills of Six Drugs with Rehmannia" were created for clinical trials. With the early successful limb salvage rate of over 80% observed in a clinical series studied at the Prince of Wales Hospital, Hong Kong, multi-directional studies on the two formulae were carried out. The aim was to find out the clinical efficacy of Formulae 1 & 2, and their component herbs, and the biological mechanism of action. A series of in-vitro, ex-vivo and in-vivo experimental models were completed for the latter purposes. / Granulation formation is an important issue essential for ulcer healing. Therefore a CRL-7522 fibroblast cell line and primary fibrobass from eight diabetic foot ulcer patients (ex-vivo) were used to detect the granulation enhancing activities of the Formulae 1 & 2 and component herbs. The two formulae and some of their component herbs viz, Radix Astragali (HQ), Radix Rehmanniae (SD) and Rhizoma Atractylodis Macrocephalae (BZ) showed significant enhancement effects on the cell viability and apparently facilitated granulation formation. Hence the Formulae 1 & 2, and the three component herbs were selected for further studies. The other nine component herbs of the formulae were found to have no significant enhancing effects on cell viability. With an established diabetic rat model (n0 STZ and n5 STZ), a piece of full-thickness skin was removed from the foot of the rat to develop a diabetic rat foot ulcer model. The ulcer area was measured by a specially designed area measuring programme, namely the Image Analytical Programme. The ulcer areas and their percentage reductions over time were recorded and analysed using statistical multilevel models with adjustments for weight, blood glucose level and the presence of extra ulcers. Results revealed that the ulcer area was significantly reduced by the Formulae 1 & 2, and one of their component herbs, Radix Rehmanniae (SD). / Lau Tai-Wai. / "February 2005." / Adviser: Ping Chung Leung. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0197. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 292-310). / 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, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.