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.

Sun Ye. / "November 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 205-225) / 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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Lau, Ka Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 223-237). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iv / Acknowledgments --- p.vi / Publication List --- p.vii / Table of Contents --- p.viii / List of Abbreviations --- p.xiv / List of Figures --- p.xvi / List of Tables --- p.xx / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Cancer --- p.1 / Chapter 1.1.1 --- Cancer in Hong Kong --- p.1 / Chapter 1.1.2 --- Different types of cancer treatments and the side effects --- p.4 / Chapter 1.1.3 --- Alternative therapies for cancer treatment --- p.6 / Chapter 1.1.3.1 --- Successful examples of anti-cancer drugs from traditional Chinese herbs --- p.7 / Chapter 1.2 --- Anti-tumor study approaches --- p.11 / Chapter 1.2.1 --- Direct cytotoxic activities --- p.12 / Chapter 1.2.2 --- Immunomodulatory activities --- p.14 / Chapter 1.2.3 --- Anti-angiogenesis activities --- p.16 / Chapter 1.3 --- Objectives of our study --- p.20 / Chapter Chapter 2 --- Background of selected Chinese herbs in our study / Chapter 2.1 --- Search for anti-tumor Chinese herbs --- p.21 / Chapter 2.1.1 --- Chinese herbs commonly used for cancer treatment --- p.21 / Chapter 2.1.2 --- Literature Search --- p.21 / Chapter 2.2 --- Results --- p.22 / Chapter 2.2.1 --- Lists of Chinese herbs from various Chinese medicine practitioners --- p.22 / Chapter 2.2.2 --- Selected traditional Chinese herbs from literature search --- p.22 / Chapter 2.2.3 --- Selected Chinese herbs for our study --- p.27 / Chapter 2.3 --- Background information of the five selected Chinese herbs --- p.28 / Chapter 2.3.1 --- Fructus Bruceae (FB) --- p.28 / Chapter 2.3.1.1 --- Traditional uses --- p.28 / Chapter 2.3.1.2 --- Previous Studies of Fructus Bruceae --- p.28 / Chapter 2.3.1.3 --- Isolated compounds of FB --- p.31 / Chapter 2.3.2 --- Cortex Phellodendri Amurensis (PA) --- p.35 / Chapter 2.3.2.1 --- Traditional uses --- p.35 / Chapter 2.3.2.2 --- Previous studies of Cortex Phellodendri Amurensis --- p.35 / Chapter 2.3.2.3 --- Previous studies of Berberine --- p.38 / Chapter 2.3.3 --- Radix et Rhizoma Asteris (RA) --- p.39 / Chapter 2.3.3.1 --- Traditional uses --- p.39 / Chapter 2.3.3.2 --- Previous Studies of Radix et Rhizoma Asteris --- p.39 / Chapter 2.3.4 --- Semen Coicis (SC) --- p.41 / Chapter 2.3.4.1 --- Traditional uses --- p.41 / Chapter 2.3.4.2 --- Previous Studies of Semen Coicis --- p.41 / Chapter 2.3.5 --- Radix Scrophulariae (RS) --- p.43 / Chapter 2.3.5.1 --- Traditional uses --- p.43 / Chapter 2.3.5.2 --- Previous Studies of Radix Scrophulariae --- p.43 / Chapter 2.4 --- Authentication of selected Chinese herbs --- p.45 / Chapter 2.4.1 --- Sources --- p.45 / Chapter 2.4.2 --- Morphological characteristics of the Chinese herbs --- p.47 / Chapter 2.4.2.1 --- Fructus Bruceae --- p.47 / Chapter 2.4.2.2 --- Cortex Phellodendri Amurensis --- p.48 / Chapter 2.4.2.3 --- Radix et Rhizoma Asteris --- p.49 / Chapter 2.4.2.4 --- Semen Coicis --- p.50 / Chapter 2.4.2.5 --- Radix Scrophulariae --- p.51 / Chapter 2.5 --- Extraction of selected Chinese herbs --- p.52 / Chapter 2.5.1 --- Materials and methods --- p.52 / Chapter 2.5.1.1 --- Preparation of aqueous extracts of selected Chinese herbs --- p.52 / Chapter 2.5.2 --- Results --- p.53 / Chapter 2.5.2.1 --- Percentage yield of aqueous extract of selected Chinese herbs --- p.53 / Chapter 2.6 --- Discussion --- p.54 / Chapter Chapter 3 --- Direct cytotoxic effect of selected Chinese herbs / Chapter 3.1 --- Background --- p.55 / Chapter 3.2 --- Materials and methods --- p.56 / Chapter 3.2.1 --- Cell cultures --- p.56 / Chapter 3.2.2 --- Determination of cell viability by MTT assay --- p.58 / Chapter 3.2.3 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.59 / Chapter 3.2.4 --- Preparation of etoposide for direct cytotoxic assay --- p.60 / Chapter 3.2.5 --- Statistical analysis --- p.61 / Chapter 3.3 --- Results --- p.62 / Chapter 3.3.1 --- Cytotoxic effects of five selected Chinese herbs on a panel of human cancer cell lines and human normal cell line --- p.62 / Chapter 3.3.2 --- Comparison of the cytotoxic effect of etoposide and the selected Chinese herbal extracts on a panel of human tumor cells --- p.72 / Chapter 3.3.3 --- Further investigations of the anti-tumor effect of PA --- p.75 / Chapter 3.3.3.1 --- Materials and methods --- p.75 / Chapter 3.3.3.1.1 --- Quantification of berberine chloride in PA aqueous extract using TLC --- p.75 / Chapter 3.3.3.1.2 --- Determination of cell viability by MTT assay --- p.76 / Chapter 3.3.3.2 --- Results --- p.76 / Chapter 3.3.3.2.1 --- Quantification of berberine chloride in PA aqueous extract using TLC --- p.76 / Chapter 3.3.3.2.2 --- Cytotoxic effect of berberine on a panel of human cancer cell lines --- p.78 / Chapter 3.4 --- Discussion --- p.80 / Chapter Chapter 4 --- Immunomodulatory effects of selected Chinese herbs / Chapter 4.1 --- Background --- p.84 / Chapter 4.2 --- Materials and methods --- p.87 / Chapter 4.2.1 --- Preparation of human peripheral blood mononuclear cells (huPBMCs) --- p.87 / Chapter 4.2.2 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.88 / Chapter 4.2.3 --- Preparation of cell mitogens --- p.88 / Chapter 4.2.4 --- Statistical analysis --- p.89 / Chapter 4.3 --- Results --- p.89 / Chapter 4.3.1 --- Mitogenic activities of the selected herbal extracts on huPBMCs --- p.89 / Chapter 4.4 --- Further investigations of the mitogenic activities of SC and RA extracts --- p.96 / Chapter 4.4.1 --- Materials and methods --- p.96 / Chapter 4.4.1.1 --- Preparation of human peripheral blood mononuclear cells (huPBMCs) --- p.96 / Chapter 4.4.1.2 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.96 / Chapter 4.4.2 --- Results --- p.97 / Chapter 4.4.2.1 --- Mitogenic effects of SC and RA aqueous extracts (in the presence of polymyxin B) --- p.97 / Chapter 4.5 --- Chemical characterization of RA aqueous extract --- p.100 / Chapter 4.5.1 --- Materials and methods --- p.100 / Chapter 4.5.1.1 --- Quantification of polysaccharide and carbohydrate contents in RA aqueous extract --- p.100 / Chapter 4.5.1.2 --- Quantification of protein content in RA aqueous extract --- p.101 / Chapter 4.5.2 --- Results --- p.103 / Chapter 4.5.2.1 --- Chemical characterization of RA aqueous extract --- p.103 / Chapter 4.6 --- Further investigations of the underlying mechanisms of the mitogenic activities of RA aqueous extract --- p.104 / Chapter 4.6.1 --- Materials and methods --- p.104 / Chapter 4.6.1.1 --- Preparation of human peripheral blood mononuclear cells (huPBMCs) --- p.104 / Chapter 4.6.1.2 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.104 / Chapter 4.6.1.3 --- Human Thl/Th2 Cytokine Cytometric Bead Array (CBA) --- p.105 / Chapter 4.6.1.4 --- Statistical analysis --- p.106 / Chapter 4.6.2 --- Results --- p.106 / Chapter 4.6.2.1 --- Effects of RA aqueous extract on productions of cytokinesin huPBMCs --- p.106 / Chapter 4.7 --- Discussion --- p.108 / Chapter Chapter 5 --- Anti-angiogenesis effects of selected Chinese herbs / Chapter 5.1 --- Background of in vivo zebrafish model --- p.112 / Chapter 5.2 --- Materials and methods --- p.117 / Chapter 5.2.1 --- Maintenance of zebrafish --- p.117 / Chapter 5.2.2 --- Collection of zebrafish embryos --- p.117 / Chapter 5.2.3 --- Zebrafish embryos treated with different herbal extracts --- p.117 / Chapter 5.2.4 --- Visual screens of zebrafish embryos using fluorescence microscopy --- p.118 / Chapter 5.2.5 --- Statistical analysis --- p.118 / Chapter 5.3 --- Results --- p.120 / Chapter 5.3.1 --- Anti-angiogenesis effect of SU5416 --- p.120 / Chapter 5.3.2 --- Anti-angiogenesis effects of selected herbal extracts on zebrafish model --- p.122 / Chapter 5.4 --- Discussion --- p.133 / Chapter Chapter 6 --- Further investigations on the anti-tumor effects of Fructus Bruceae and its sub-fractions / Chapter 6.1 --- Introduction --- p.136 / Chapter 6.2 --- Solvent partition of FB aqueous extract --- p.138 / Chapter 6.2.1 --- Materials and methods --- p.138 / Chapter 6.2.1.1 --- Solvent partition --- p.138 / Chapter 6.2.1.2 --- Thin layer chromatography of FB fractions --- p.138 / Chapter 6.2.2 --- Results --- p.139 / Chapter 6.2.2.1 --- Percentage yield of different fractions of FB aqueous extract --- p.139 / Chapter 6.2.2.2 --- Thin layer chromatography of FB fractions --- p.140 / Chapter 6.3 --- Investigations of the anti-tumor activities of FBW fraction --- p.141 / Chapter 6.3.1 --- Materials and methods --- p.141 / Chapter 6.3.1.1 --- Cell cultures --- p.141 / Chapter 6.3.1.2 --- Determination of cell viability by MTT assay --- p.141 / Chapter 6.3.1.3 --- Preparation of human peripheral blood mononuclear cells (huPBMCs) --- p.141 / Chapter 6.3.1.4 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.141 / Chapter 6.3.1.5 --- Statistical analysis --- p.141 / Chapter 6.3.2 --- Results --- p.142 / Chapter 6.3.2.1 --- Cytotoxic effects of FBW on a panel of human cancer cells and human normal cells --- p.142 / Chapter 6.3.2.2 --- Mitogenic activities of FBW fraction on huPBMCs --- p.145 / Chapter 6.4 --- Chemical characterizations of FB aqueous extract and FBW fraction --- p.147 / Chapter 6.4.1 --- Materials and methods --- p.147 / Chapter 6.4.2 --- Results --- p.147 / Chapter 6.5 --- Bioassay guided fractionation of FBW --- p.149 / Chapter 6.5.1 --- Fractionation using macroporous resin column (D101) --- p.149 / Chapter 6.5.2 --- Investigations of the anti-tumor effects of the sub-fractions of FBW --- p.151 / Chapter 6.5.2.1 --- Direct cytotoxic effects of FBW sub-fractions on NB-4 cells and human normal cells --- p.151 / Chapter 6.5.2.2 --- Immunomodulatory effects of FBW-DH sub-fraction --- p.154 / Chapter 6.5.3 --- Fractionation using ethanol precipitation --- p.155 / Chapter 6.5.3.1 --- Chemical characterization of sub-fractions of FBW-DH --- p.156 / Chapter 6.5.3.2 --- "Direct cytotoxic effects of 50P, 80P and 80S on NB-4 cells and human normal cells" --- p.159 / Chapter 6.5.3.2.1 --- DNA agarose gel electrophoresis --- p.163 / Chapter 6.5.3.2.2 --- Cell death detection ELISA --- p.166 / Chapter 6.5.3.2.3 --- ELISA of apoptotic related proteins --- p.168 / Chapter 6.5.3.2.4 --- Telomerase PCR ELISA --- p.176 / Chapter 6.5.3.3 --- "Immunomodulatory effects of 50P, 80P and 80S" --- p.178 / Chapter 6.5.3.3.1 --- Human Thl/Th2 cytokine cytometric bead array (CBA) --- p.180 / Chapter 6.5.3.3.2 --- Limulus Amebocyte Lysate assay --- p.183 / Chapter 6.5.3.4 --- "Anti-angiogenic effects of 50P, 80P and 80S" --- p.184 / Chapter 6.5.3.5 --- Liquid chromatography mass spectrometry (LCMS) analysis of 50P --- p.192 / Chapter 6.6 --- Discussion --- p.194 / Chapter Chapter 7 --- General discussions and conclusions / Chapter 7.1 --- Anti-tumor activities of five selected Chinese herbs --- p.202 / Chapter 7.2 --- Significance of the present study --- p.213 / Chapter 7.3 --- Limitations of our study --- p.214 / Chapter 7.4 --- Future work --- p.215 / Appendices / Appendix I Phenol-sulphuric acid spectrophotometric assay --- p.216 / Appendix II Bradford assay --- p.217 / Appendix III Calibration curves of cytokines in CBA assay --- p.218 / Appendix IV Endotoxin standard curve --- p.220 / Appendix V LCMS data of two chemical markers of FB --- p.221 / Bibliography --- p.223

黑色素生成是為了保護皮膚細胞免受紫外光傷害的一個生化過程。在這過程中，黑色素在人類表皮底層的黑色素細胞的黑色素體內產生。該過程可以被基因，荷爾蒙或環境因素所影響。黑色素的製造量是依賴速度限制酶酪氨酸酶的活性，黑色素體的數量和大小，黑色素體通過黑色素細胞的偽足傳送致角質細胞的速度及黑色素體在角質細胞內的分佈。這些細胞過程會受皮膚顏色或紫外光曝光量的變化而影響。當黑色素的產生超過黑色素的降解，黑色素沉著毛病便出現。根據不同的皮膚類型，年齡組別及累積紫外光曝光程度而引發雀斑或黃褐斑的形成。很多治療方法市面上能夠提供，它們包括人工合成化粧品，激光，整容手術等。這些治療方法通常會產生副作用及蘊藏高風險。因此從天然物質裏尋找治療藥物便成了美容學的一個新的研究方向。在這研究裏，十種草本植物就從自古以來用作治療黑色素沉著毛病的傅統中藥中被挑選出來。那些草本植物被四種擁有不同極性的溶劑提取。小鼠黑色素細胞被用以篩選提取物的降黑色素能力。結果發現當歸的正己烷及二氯甲烷的提取物有正面效用。當歸的化學成份4-乙基間苯二酚、4-乙基苯酚及1-十四烷醇也能降低小鼠黑色素細胞的黑色素量。數種生化技術繼而被應用作研究有效化學物的藥理。他們包括西方墨點法、環磷酸腺苷測試、蛋白激酶A活性測試及酪氨酸酶活性測試。 / Melanogenesis is a biochemical process designated for protecting skin cells from ultraviolet (UV)-induced damage. During the process, melanin is produced in the melanosomes of the melanocytes located at the basal epidermis of human. The process could be affected by genetic, hormonal or environmental factors. Amount of melanin synthesized depending on the activity of the rate-limiting enzyme tyrosinase, number and size of melanosomes, the transfer rate of melanosomes to keratinocytes through the melanocyte dendritic projections and the distribution pattern of melanosomes within keratinocytes. These cellular processes are influenced by variations in skin color or UV exposure amount. When melanin synthesis exceeds melanin degradation, hyperpigmentation disorder arises. This lead to the formation of freckles or chloasma according to different skin types, age groups and degree of cumulative UV exposure. A number of treatments are commercially available, they include applying synthetic cosmetics, laser, plastic surgery, etc. These treatments usually produce side-effects and possess high risk. Therefore, searching for therapeutic agents from natural compounds has become a new research direction in cosmetology. In this study, ten herbs were chosen from traditional Chinese medicine (TCM) which had been applied for treating hyperpigmentation. The herbs were extracted by four solvents with different polarity. The extracts were screened for their hypopigmentary ability by using melan-a cells. It was found that the hexane and dichloromethane extracts of Angelica sinensis possessed positive effects. 4-ethylresorcinol, 4-ethylphenol and 1-tetradecanol, the chemical constituents of A. sinensis, also attenuated melanin amount in melan-a cells. Moreover, several biochemical techniques were utilized to study the pharmaceutical mechanisms of the potent compounds. They include Western blot, cyclic adenosine monophosphate (cAMP) assay, protein kinase A (PKA) activity assay and tyrosinase activity assay. / Detailed summary in vernacular field only. / Lam, Rosanna Yen Yen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 127-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / Chinese Abstract --- p.iii / Acknowledgements --- p.iv / List of Publications --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Demand of cosmetics --- p.1 / Chapter 1.2 --- Skin structures and functions --- p.1 / Chapter 1.2.1 --- Epidermis --- p.3 / Chapter 1.2.1.1 --- Stratum corneum --- p.4 / Chapter 1.2.1.2 --- Stratum granulosum --- p.4 / Chapter 1.2.1.3 --- Stratum spinosum --- p.4 / Chapter 1.2.1.4 --- Stratum basale --- p.4 / Chapter 1.2.2 --- Dermis --- p.5 / Chapter 1.2.3 --- Hypodermis --- p.5 / Chapter 1.3 --- Sun irradiation --- p.5 / Chapter 1.4 --- Variety of skin types --- p.6 / Chapter 1.5 --- Biochemical reactions within melanocyte --- p.7 / Chapter 1.6 --- Pigmentation disorder --- p.14 / Chapter 1.7 --- From the view of traditional Chinese medicine --- p.16 / Chapter 1.8 --- Treatments available for hyperpigmentation --- p.18 / Chapter 1.9 --- Aims of study and application of strategies --- p.19 / Chapter Chapter 2 --- Investigation of the inhibitory effect of herbal extracts and their constituent compounds on melanin synthesis --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials and methods --- p.21 / Chapter 2.2.1 --- Materials --- p.21 / Chapter 2.2.2 --- Herbal extraction --- p.23 / Chapter 2.2.3 --- Cell culture --- p.25 / Chapter 2.2.4 --- Growth curve and melanin production curve --- p.25 / Chapter 2.2.5 --- SRB assay --- p.26 / Chapter 2.2.6 --- Calibration curve for SRB assay --- p.27 / Chapter 2.2.7 --- Measurement of melanin production --- p.27 / Chapter 2.2.8 --- Calibration curve for melanin production assay --- p.28 / Chapter 2.2.9 --- Statistical analysis --- p.28 / Chapter 2.3 --- Results --- p.29 / Chapter 2.3.1 --- Growth curve and melanin production curve for assay development --- p.29 / Chapter 2.3.2 --- Calibration curves of SRB assay and melanin production assay --- p.32 / Chapter 2.3.3 --- Hypopigmentary effect of 40 herbal extracts --- p.35 / Chapter 2.3.4 --- Hypopigmentary effects of chemical components of A. sinensis --- p.41 / Chapter 2.4 --- Discussion --- p.49 / Chapter Chapter 3 --- Study of the effect of potential compounds on melanogenic protein expression by Western blot --- p.54 / Chapter 3.1 --- Introduction --- p.54 / Chapter 3.2 --- Materials and methods --- p.56 / Chapter 3.2.1 --- Materials --- p.56 / Chapter 3.2.2 --- Cell culture --- p.56 / Chapter 3.2.3 --- Preparation of cell lysates --- p.57 / Chapter 3.2.4 --- Protein assay --- p.57 / Chapter 3.2.5 --- SDS-PAGE and membrane transfer --- p.58 / Chapter 3.2.6 --- Washing of blotted antibodies and film exposure --- p.59 / Chapter 3.3 --- Results --- p.61 / Chapter 3.4 --- Discussion --- p.70 / Chapter Chapter 4 --- Study of the effect of potential compounds on melanogenic gene expression by RT-PCR --- p.76 / Chapter 4.1 --- Introduction --- p.76 / Chapter 4.2 --- Materials and methods --- p.76 / Chapter 4.2.1 --- Materials --- p.77 / Chapter 4.2.2 --- Cell culture --- p.77 / Chapter 4.2.3 --- RNA extraction --- p.78 / Chapter 4.2.4 --- cDNA synthesis --- p.78 / Chapter 4.2.5 --- PCR --- p.80 / Chapter 4.3 --- Results --- p.83 / Chapter 4.4 --- Discussion --- p.85 / Chapter Chapter 5 --- Study of the effect of potential compounds on cAMP level by EIA --- p.85 / Chapter 5.1 --- Introduction --- p.86 / Chapter 5.2 --- Materials and methods --- p.86 / Chapter 5.2.1 --- Materials --- p.86 / Chapter 5.2.2 --- Cell culture --- p.86 / Chapter 5.2.3 --- Preparation of cell lysates --- p.86 / Chapter 5.2.4 --- Protein assay --- p.87 / Chapter 5.2.5 --- The cAMP assay --- p.88 / Chapter 5.2.6 --- Preparation of cAMP calibration curve --- p.88 / Chapter 5.2.7 --- Calculation --- p.89 / Chapter 5.2.8 --- Statistical analysis --- p.89 / Chapter 5.3 --- Results --- p.90 / Chapter 5.4 --- Discussion --- p.94 / Chapter Chapter 6 --- Study of the effect of potential compounds on PKA activity by PKA activity assay --- p.96 / Chapter 6.1 --- Introduction --- p.96 / Chapter 6.2 --- Materials and methods --- p.96 / Chapter 6.2.1 --- Materials --- p.97 / Chapter 6.2.2 --- Cell culture --- p.97 / Chapter 6.2.3 --- Preparation of cell lysates --- p.98 / Chapter 6.2.4 --- Protein assay --- p.98 / Chapter 6.2.5 --- The PKA kinase activity assay --- p.100 / Chapter 6.2.6 --- Calculation --- p.100 / Chapter 6.2.7 --- Statistical analysis --- p.100 / Chapter 6.3 --- Results --- p.101 / Chapter 6.4 --- Discussion --- p.104 / Chapter Chapter 7 --- Study of the effect of potential compounds on tyrosinase activity by enzyme inhibition assay --- p.107 / Chapter 7.1 --- Introduction --- p.107 / Chapter 7.2 --- Materials and methods --- p.108 / Chapter 7.2.1 --- Materials --- p.108 / Chapter 7.2.2 --- Assay development for mushroom tyrosinase --- p.109 / Chapter 7.2.3 --- Mushroom tyrosinase inhibition assay --- p.109 / Chapter 7.2.4 --- Cell culture --- p.110 / Chapter 7.2.5 --- Preparation of cellular tyrosinase --- p.110 / Chapter 7.2.6 --- Protein assay --- p.111 / Chapter 7.2.7 --- Cellular tyrosinase inhibition assay --- p.111 / Chapter 7.2.8 --- Calculation --- p.112 / Chapter 7.2.9 --- Statistical analysis --- p.112 / Chapter 7.3 --- Results --- p.113 / Chapter 7.4 --- Discussion --- p.120 / Chapter Chapter 8 --- General discussion --- p.123 / References --- p.127

In a mouse model of pulmonary thromboembolism induced by a collagen-adrenaline mixture, the SFE extract and ligustilide reduced the paralysis-death ratio, and the anti-thrombotic response of senkyunolide A was more pronounced. The effect of BDPH was not significant. Neither the SFE extract nor the three phthalides prolonged bleeding time in tail-transected mice. / In a rat myocardial ischemia-reperfusion model involving coronary artery ligation, 7-day pre-treatment with the SFE extract and ligustilide reduced ventricular arrhythmias in isolated hearts. BDPH and senkyunolide A were without significant effects. / In rat platelet-rich plasma, platelet aggregation induced by collagen and U46619 but not by adenosine diphosphate was inhibited by the SFE extract. Ligustilide inhibited the responses of all three agonists, while BDPH and senkyunolide A inhibited the collagen response only. / Raw Rhizoma Chuanxiong herb and its crude extract as obtained by supercritical fluid extraction (SFE) comprised mainly phthalides. The SFE extract and three representative phthalides, butylidenephthalide (BDPH), ligustilide and senkyunolide A, were studied on vasorelaxation, myocardial ischemia, platelet aggregation and thrombosis. The mechanisms underlying BDPH-mediated vasorelaxation were also explored. / Rhizoma Chuanxiong, the dried rhizome of Ligusticum chuanxiong Hort., is a common traditional Chinese medicine used for the treatment of cardiovascular diseases. Surprisingly, the scientific basis of its action is poorly understood. The current study aims to establish the pharmacological basis of the cardiovascular effects of Rhizoma Chuanxiong and its active constituents by examining their effects in several cardiovascular domains. / The current study demonstrated various cardiovascular actions of Rhizoma Chuanxiong, and thereby established the pharmacological basis of the effects of the herb. Phthalides, in particular BDPH, ligustilide and senkyunolide A, were important contributors to such actions. Future investigation of the SFE extract and/or individual phthalides related to the progression from in vitro and in vivo effectiveness to clinical efficacy is much anticipated. / The SFE extract, BDPH, ligustilide and senkyunolide A produced vasorelaxation on isolated preparations of rat aorta, rat saphenous vein and pig coronary artery. BDPH-mediated relaxation appeared to involve both extracellular Ca 2+-dependent (L-type voltage-operated, receptor-operated and store-operated Ca2+ channels) and independent (NO modulation, Ca2+ release from internal stores and Ca2+ desensitization) mechanisms. BDPH was also observed to augment relaxation induced by sodium nitroprusside and forskolin through mechanisms that remain undefined. / Chan Sun Kin Sunny. / "July 2005." / Advisers: G. Lin; R. L. Jones. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1575. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 190-209). / 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.

differential scanning calorimetric profile and the generation of much less intense and broader peaks in the powder X-ray diffraction pattern compared to beta-CD. FTIR analysis revealed significant physical interactions between CX oil and beta-CD in the granules, possibly due to complexation. Results from phase solubility measurements and proton nuclear magnetic resonance ( 1H-NMR) analysis of pure 3-butylidenephthalide (3-BDPH), a representative CX component, lend some support for the formation of a 1:1 stoichiometric inclusion complex between 3-BDPH and beta-CD. / Rhizoma chuanxiong (CX), the dried rhizome of Ligusticum Chuangxiong Hort. (Umbelliferae), has been extensively used in mainland China as a traditional herbal medicine for treating cardio-/cerebrovascular diseases and gynecological disorders. However, the active components in CX, which are predominantly essential oils, generally exhibit poor stability (mostly photo-oxidation), high volatility, low aqueous solubility, and extensive gut/hepatic metabolism, all of which can significantly reduce their oral bioavailability and therapeutic efficacy. The present project has investigated the feasibility of utilizing three formulation approaches to circumvent the aforementioned problems associated with the peroral delivery of CX (as crude oil mixture or individual components). / The first approach involved inclusion of CX oil in beta-cyclodextrin (beta-CD) as solid granules using a coprecipitation method optimized with the aid of an orthogonal study design. The resulting CX oil granules were colorless and odorless with a median particle size of 11.38mum; were stable to heat, light and moisture, and readily soluble in simulated gastric and intestinal fluids. The granules were largely amorphous, as evidenced by an absence of the melting endotherm for beta-CD in the formulation could be largely explicated by the complexation behavior and hydration properties of the two polymers blended in different weight percentages, as substantiated by turbidity measurement, viscosity determination and FTIR analysis of the pure polymer mixtures as well as swelling measurement of the formulated tablets. The sustained release behavior of 3-BDPH from the tablet was dependent on the relative proportion of the two polymers present, and could be similarly explained by the changes in hydration and complexation behavior of the polymers during the penetration of aqueous fluid into the tablet matrix. / The second approach involved incorporation of CX oil into surfactant micelles and liquid crystals as a self-emulsifying drug delivery system (SEDDS). An optimal formulation was developed through a judicial choice of excipients (lipids and surfactants/cosurfactant) and their proper combination in the correct proportions, as determined by the spontaneity of the emulsification process and the change in emulsion droplet size. The formulation was readily dispersible in water upon mild agitation, free from unpleasant odor, and stable in soft gelatin capsules for a storage period of at least 12 months under ambient condition. The optimal utilization of the lipid and surfactant blends in defined proportions in the formulation was further substantiated by interfacial tension determination and equilibrium phase analysis. / The third approach involved formulation of 3-BDPH (or crude CX oil) into a sustained-release buccoadhesive tablet, based on a systematic evaluation of the adhesive properties of two polymers (Carbopol 974P and hydroxypropyl methylcellulose K4M) used in the formulation. The adhesive properties of the formulation could be largely explicated by the complexation behavior and hydration properties of the two polymers blended in different weight percentages, as substantiated by turbidity measurement, viscosity determination and FTIR analysis of the pure polymer mixtures as well as swelling measurement of the formulated tablets. The sustained release behavior of 3-BDPH from the tablet was dependent on the relative proportion of the two polymers present, and could be similarly explained by the changes in hydration and complexation behavior of the polymers during the penetration of aqueous fluid into the tablet matrix. / Gao Yuan. / "April 2005." / Adviser: Albert H. L. Chow. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1585. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 193-223). / 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.

Besides the molecular mechanisms regulating activation of FLS mentioned above, we also investigated anti-inflammatory activities of Chinese herbal medicine sinomenine and Liang Miao San on activated human FLS in RA. Sinomenine, an alkaloid isolated from the root of Sinomenium acutum, has been used to alleviate the symptoms of rheumatic diseases. Liang Miao San (LMS), composed of the herbs Rhizoma Atractylodis (Cangzhu) and Cotex Phellodendri (Huangbai), is another traditional Chinese medicine formula for RA treatment. Since the potential anti-inflammatory activities of sinomenine and LMS have been demonstrated, we investigated the in vitro anti-inflammatory effects of sinomenine and LMS on inflammatory cytokine TNF-alpha activation of human normal and RA-FLS and the underlying intracellular mechanisms. In the present study, sinomenine was found to significantly inhibit TNF-alpha induced cell surface expression of VCAM-1 and release of inflammatory cytokine and chemokine IL-6, CCL2 and CXCL8 from both normal and RA-FLS (all p < 0.05). Our results provide a new insight into the differential anti-inflammatory activities of sinomenine and LMS through the suppression of TNF-alpha activated FLS by modulating distinct intracellular signaling pathways in RA, and help to provide a biochemical basis for the development of a cost-effective human synoviocyte model for the future screening of traditional Chinese medicine (TCM) possessing potential anti-rheumatic activities. (Abstract shortened by UMI.) / IL-27, a novel member of the IL-12 family that is produced early by antigen-presenting cells (APCs), can promote T cell proliferation as well as the production of interferon-gamma by naive T lymphocytes. Recent studies have found that elevated expression of IL-27 has been detected in the synovial membranes and fluid of RA. Herein we investigated the in vitro effects of IL-27, alone or in combination with inflammatory cytokine TNF-alpha or IL-Ibeta on the pro-inflammatory activation of human primary FLS isolated from RA patients and normal control subjects, and the underlying intracellular signaling molecules were also studied. We found that the plasma concentration of IL-27 in RA patients (n=112) was significantly higher than that in control subjects (n=46). Both normal and RA-FLS constitutively express functional IL-27 receptor heterodimer, gp130 and WSX-1, with more potent IL-27-mediated activation of signal transducers and activators of transcription (STAT)1 in RA-FLS. IL-27 was found to induce significantly higher cell surface expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 and release of inflammatory cytokine IL-6, chemokine CCL2, CXCL9, CXCL10 and matrix metalloproteinase (MMP)-1 of RA-FLS than that of normal FLS (all p < 0.05). The above findings therefore provide a new insight into the IL-27-activated immunopathological mechanisms mediated by distinct intracellular signal transductions in joint inflammation of RA and may have important therapeutic implications. / In the present study, we have investigated the mechanisms of the activation of human fibroblast-like synoviocytes (FLS) induced by various stimuli including interleukin (IL)-27, tumor necrosis factor (TNF)-alpha and IL-beta. The activation of human FLS was studied in terms of the release of cytokines and chemokines and the expression of adhesion molecules. / We investigated the in vitro effects of uric acid crystals, alone or in combination with inflammatory cytokine TNF-alpha or IL-beta on the pro-inflammatory activation of human FLS from RA patients and normal control subjects, and the underlying intracellular signaling molecules were also determined. In the present study, uric acid crystals were found to result in a significant increase of inflammatory cytokine IL-6, chemokine CXCL8 and MMP-1 from both normal and RA-FLS (all p < 0.05). Moreover, additive or synergistic effect was observed in the combined treatment of uric acid crystals and TNF-alpha or IL-1beta on the release of IL-6, CXCL8 and MMP-1 from both normal and RA-FLS. Further investigations showed that the release of inflammatory cytokine, chemokine and matrix metalloproteinase stimulated by uric acid crystals was differentially regulated by intracellular activation of extracellular signal-regulated kinase (ERK) and JNK pathways. Our results therefore provide a new insight into the endogenous danger signal uric acid crystals-activated immunopathological mechanisms mediated by distinct intracellular signal transductions in joint inflammation, and also provide biochemical basis for the development of new modality for inflammatory rheumatic diseases. / Chen, Dapeng. / Adviser: Wong Chun Kwok. / Source: Dissertation Abstracts International, Volume: 73-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 203-240). / 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. / Abstract also in Chinese.

丹参是一种著名的传统中药，富含丹酚酸和丹参酮。其中，丹参酮的潜在抗肿瘤作用近年来引起众多关注。本研究评价了主要的丹参酮及其衍生物对结肠癌细胞的细胞毒性。结果显示DHTS具有最强的抗结肠癌活性和显著的肿瘤特异性细胞毒性，其细胞毒性主要由于凋亡诱导而不是引起坏死。初步的构效关系分析提示丹参酮母环结构中的A环和B环增加的离域性有助于提高其对结肠癌细胞的细胞毒性，而非二维结构及较小的D环也是进行结构改造的可能方向。 / 基于以上发现，本研究进一步探讨了DHTS的体内外抗肿瘤活性及内在机制。本研究发现DHTS的促凋亡活性并不依赖于p53的表达，而依赖于caspase活性及线粒体介导的细胞质中氧自由基 ROS及钙离子的聚集。DHTS可引起浓度及时间依赖caspase-9/-3/-7的活化而并未显著引起caspase-8的活化，这一现象发生于同样以浓度及时间依赖方式进行的线粒体中cytochrome c及AIF转位之后。在DHTS诱导的结肠癌细胞凋亡中，cytochrome c及caspase介导的凋亡通路及AIF介导的凋亡通路均被激活并显示出两条通路之间的交叉调控。 / 此外，线粒体在DHTS的促凋亡活性中的作用在本研究中被深入探讨。本研究发现线粒体可能是DHTS的一个直接靶点, 而氧化磷酸化复合体III则更可能是其作用位点。DHTS可以引起迅速而明显的线粒体功能障碍，随之引起细胞质中大量的氧自由基及钙离子聚集，诱导凋亡的产生。 / 与体外结果一致，本研究证实了DHTS对免疫缺陷小鼠中的结肠癌移植廇也具有明显的抗肿瘤作用。与溶媒对照组比较，DHTS治疗组中移植廇的增长显著被减缓，在治疗终点时的廇体积与重量也显著被降低。TUNEL检测确认DHTS诱导移植廇中癌细胞的显著凋亡。免疫荧光检测也发现DHTS诱导caspase-3及caspase-7在移植廇中癌细胞的明显活化。 / 综上所述，本研究提供了丹参酮抗结肠癌活性的一些初步构效关系的信息，为提高丹参酮抗结肠癌活性的结构改造提供一定的参考。更重要的是，本研究证明了DHTS的体内外抗结肠癌活性并探讨了其作用机制及可能靶点，为DHTS作为新的应用于抗结肠癌药物或辅助治疗用药提供了临床前研究证据。 / Salvia miltiorrhiza Bunge, also known as Danshen, rich in phenolic acid and tanshinones, has been widely used to treat various kinds of diseases including heart diseases and hepatitis in China with minimal side effects. Among the tanshinones, tanshinone I, tanshinone IIA, cryptotanshinone and dihydrotanshinone I are the major bioactive constituents in this herb. In this study, the anti-colon cancer potential of five tanshinones and six derivatives of tanshinone IIA were evaluated in several colon cancer cell lines. It was found that apoptosis but not necrosis contributed significantly to the cytotoxicity of the tanshinones. Dihydrotanshinone I (DHTS) was confirmed to be the most potent and selective anti-cancer compound among the tanshinones tested in this study. Preliminary SAR (structure activity relationship) of tanshinones reveals that the increase of delocalizability of A and B rings in the chemical structure of the tanshinones enhances their cytotoxicity on cancer cells, while compounds with a non-planar and small sized D ring region are better choices for anti-cancer effect. / The underlying mechanisms of the anti-colon cancer activity of DHTS were further studied. It was found that apoptosis induced by DHTS was p53 independent but caspase dependent, which was closely related to intracellular accumulation of ROS (reactive oxidant stress) and calcium mediated by mitochondria. A concentration- and time-dependent activation of caspase-9,-3,-7 but not caspase-8 by DHTS in HCT116 cells was detected after the translocation of cytochrome c and AIF (apoptosis inducing factor) from mitochondria. In this process, the crosstalk between the caspase-dependent and caspase-independent pathways was firstly shown in the apoptotic mechanism of DHTS. To this end, the release of cytochrome c happened first and the translocation of apoptosis inducing factor (AIF) was prevented by a pan caspase inhibitor. In the meantime, the release of cytochrome c and activation of caspase-9 and PARP (poly-ADP-ribose polymerase) cleavage were decreased after AIF knockdown. Especially, mitochondrion was suggested to be the direct target of DHTS and OXPHOS complex III but not OXPHOS complex I was probably the acting site of DHTS. / In accordance with the results obtained in vitro, the potential anti-colon cancer activity of DHTS was also observed in nude mice with xenograft tumors and the compound did not produce any observable systemic toxicity. DHTS efficiently delayed tumor growth by decreasing the tumor size and weight through the induction of apoptosis in cancer cells but not by inhibition of cell proliferation. In the same tissues, a distinct activation of caspase-3 and caspase-7 in tumor cells was also detected by immunofluorescence assay. / Collectively, the present study provides preliminary information about the SAR of the anti-colon cancer activity for tanshinones. It also confirms that DHTS is a promising compound for anti-cancer action both in vitro and in vivo. In addition, this study gives us a better understanding regarding the mechanisms of how DHTS induces apoptosis in cancer cells. All these findings could provide solid pre-clinical evidence to propel the development and application of DHTS and perhaps its derivatives as novel therapeutic or adjuvant agents for the treatment of colon cancer. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Wang, Lin. / Thesis (Ph.D.) Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 122-132). / Abstracts also in Chinese. / Wang, Lin.

Chan Chun-pong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 171-176). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iii / Abbreviation --- p.iv / Table of contents --- p.v / List of figures --- p.xi / List of tables --- p.xvi / Chapter Chapter 1 --- introduction / Chapter 1.1 --- Traditional Chinese medicines (TCMs) --- p.2 / Chapter 1.2 --- Liver disorders in Asia Pacific region --- p.3 / Chapter 1.3 --- Classification of liver disorders --- p.7 / Chapter 1.3.1 --- Hepatitis --- p.8 / Chapter 1.3.1.1 --- Hepatitis A virus infection --- p.8 / Chapter 1.3.1.2 --- Hepatitis B virus infection --- p.9 / Chapter 1.3.1.3 --- Hepatitis C virus infection --- p.11 / Chapter 1.3.1.4 --- Hepatitis D virus infection --- p.12 / Chapter 1.3.1.5 --- Hepatitis E virus infection --- p.12 / Chapter 1.3.2 --- Cancer of the liver --- p.13 / Chapter 1.3.2.1 --- Hepatocellular carcinoma --- p.13 / Chapter 1.3.2.2 --- Cholangiocarcinoma --- p.14 / Chapter 1.3.2.3 --- Metastatic liver cancer --- p.14 / Chapter 1.4 --- Conventional treatment of liver disorders --- p.14 / Chapter 1.5 --- Role of traditional Chinese medicines in hepatoprotective functions --- p.16 / Chapter 1.5.1 --- Abri Herba (Abrus Cantoniensis Hance) --- p.17 / Chapter 1.5.2 --- Rhizoma Coptidis (Coptidis chinensis Franch) --- p.18 / Chapter 1.5.3 --- Fructus Forsythia (Forsythia suspense (Thunb) Vahl) --- p.22 / Chapter 1.6 --- Molecular studies of hepatoprotective effects of TCMs --- p.26 / Chapter 1.6.1 --- Roles of detoxofication enzymes in hepatoprotection --- p.27 / Chapter 1.6.2 --- Studies of growth-related genes in cell cycle control --- p.29 / Chapter 1.7 --- Aims of project --- p.32 / Chapter 1.8 --- Application of the project --- p.33 / Chapter Chapter 2 --- Methods and materials --- p.34 / Chapter 2.1 --- Screening of traditional Chinese medicines --- p.35 / Chapter 2.2 --- Preparation of TCMs --- p.35 / Chapter 2.2.1 --- Preparation of aqueous extracts of TCMs --- p.35 / Chapter 2.2.2 --- Preparation of active components of TCMs --- p.36 / Chapter 2.3 --- In vitro assays --- p.40 / Chapter 2.3.1 --- Cell culture --- p.40 / Chapter 2.3.2 --- Cytotoxicity test --- p.40 / Chapter 2.4 --- Screening of expressed gene induced by TCMs --- p.41 / Chapter 2.4.1 --- RNA preparation --- p.41 / Chapter 2.4.2 --- cDNA array hybridization --- p.42 / Chapter 2.4.3 --- Reverse Transcription --- p.43 / Chapter 2.5 --- Confirmation of expressed genes induced by TCMs --- p.44 / Chapter 2.5.1 --- Semi-quantitative PCR analysis --- p.44 / Chapter 2.5.2 --- Northern blot analysis --- p.46 / Chapter 2.6 --- Studies of effects of TCMs in gene expression --- p.47 / Chapter 2.6.1 --- Dosage-course study --- p.47 / Chapter 2.6.2 --- Time-course study --- p.48 / Chapter Chapter 3 --- Results --- p.50 / Chapter 3.1 --- "Cytotoxicity test of A.H., R.C. and F.F" --- p.51 / Chapter 3.2 --- "Molecular screening of expressed gene induced by A.H., R.C., F.F" --- p.58 / Chapter 3.3 --- Confirmation of expressed gene using semi-quantitative RT- PCR --- p.70 / Chapter 3.3.1 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.70 / Chapter 3.3.2 --- Dosage-course and time-course studies of R.C. using RT- PCR --- p.94 / Chapter 3.3.3 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.113 / Chapter 3.4 --- Confirmation of expressed gene using northern blot anaylsis --- p.118 / Chapter 3.4.1 --- Dosage-course and time-course studies of effects of A.H. and L- abrine in Northern blot analysis --- p.118 / Chapter 3.4.2 --- Dosage-course and time-course studies of effects of R.C. and berberine in Northern blot analysis --- p.129 / Chapter 3.4.3 --- Dosage-course and time-course studies of effects of F.Fin Northern blot analysis --- p.147 / Chapter Chapter 4 --- Discussion --- p.152 / Chapter 4.1 --- "Roles of A.H., R.C. and F.F. in treatment and prevention of liver disorders" --- p.153 / Chapter 4.2 --- "Cytotoxicity effect A.H., R.C., and F.F. in liver cells" --- p.153 / Chapter 4.3 --- Effects of herbal medicines on the transcription of mRNA in liver cells --- p.155 / Chapter 4.3.1 --- Effects of treatment of A.H. in liver at transcriptional level … --- p.155 / Chapter 4.3.2 --- Effects of treatment of R.C. in liver at transcriptional level … --- p.156 / Chapter 4.3.3 --- Effects of treatment of R.C. in liver at transcriptional level --- p.157 / Chapter 4.4 --- Comparison of results of RT-PCR and Northern blot analysis --- p.157 / Chapter 4.4.1 --- Comparison of the effects of time and dosage-course studies of DTD expression induced by A.H. and L-abrine --- p.157 / Chapter 4.4.2 --- Comparison of the effects of time and dosage-course studies of p21;cip;waf1 expression induced by A.H. and L-abrine --- p.158 / Chapter 4.4.3 --- Comparison of the effects of time and dosage-course studies of c-myc responsive protein; rcl expression induced by R.C. and berberine --- p.159 / Chapter 4.4.4 --- Comparison of the effects of time and dosage-course studies of GST Ya expression induced by R.C. and berberine --- p.160 / Chapter 4.4.5 --- Comparison of the effects of time and dosage-course studies of GST 7-7 expression induced by F.F --- p.160 / Chapter 4.5 --- Biochemical significance of genes induced by hepatoprotective TCMs --- p.161 / Chapter 4.5.1 --- Roles of significant expression of detoxifying enzymes induced by TCMs in liver cells --- p.161 / Chapter 4.5.2 --- Roles of induction of growth-related c-myc responsive protein; rcl in R.C. treated liver cells --- p.167 / Chapter 4.5.3 --- Roles of increased p21;cip;waf1 expression in A.H. treated liver cells --- p.168 / Chapter 4.6 --- Conclusion --- p.169

Xian, Jiawen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 201-230). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.