銀屑病是一種免疫相關的慢性炎症性皮膚病，其發病率約占世界人口的1-3%，而現今仍然缺乏有效安全的根治方法。國內外使用中草藥治療銀屑病取得較好的療效，但目前缺少對其進行系統研究和開發。我們研究小組之前對61種常用治療銀屑病中藥進行篩選, 發現中藥茜草根和南蛇藤的乙醇提取物具有強大的抑制表皮細胞增生的作用，本博士研究課題的目的是確定新的安全有效的用于治療銀屑病的中藥化學成分, 並闡明其作用機制。 / 本研究篩選了28種存在于這兩種中藥中的化學單體成分，采用體外培養永生化的人類皮膚良性角質形成細胞株HaCaT, 應用MTT法, 繪制細胞生長曲線，獲得抑制50%細胞生長所需藥物濃度(IC50)。實驗結果發現1-羟基-3-甲基蒽醌（HMA）， 1，4-二氨基-2，3-（2-苯氧基乙氧基）蒽醌 （DBA）和南蛇藤表現了強大的抗表細胞生長作用，其48小時培養後的IC50分別爲17.9，15.8，1.1 μM. 值得一提的是這些化合物對正常人表皮角質細胞HEK和人類成纖維細胞Hs68只有相對輕微細胞毒性。 / 隨後進行的機理研究，通過熒光染色，DNA凝膠電泳，細胞周期檢測，流式細胞計檢測及Western blot 分析結果表明， HMA和南蛇藤素是通過誘導細胞凋亡作用抑制HaCaT細胞生長。其中南蛇藤素通過線粒體凋亡和死亡受體介導的兩種通路誘導細胞凋亡， 其誘導細胞凋亡作用與其抑制核因子-κB在HaCaT細胞中的表達和活化有關。 / 另一方面，DBA 抑制人體表皮角質細胞生長的作用機理在于其對角質細胞終末分化的誘導作用。DBA與HaCaT和HEK細胞共同培養96小時後，能顯著促進細胞角質化外膜形成，同時上調角蛋白K1/10，人體套膜蛋白，轉谷氨酰胺酶-1表達和下調角蛋白K5/14表達。而利用小鼠尾部鱗片表皮模型對HMA的外用制劑進行測試，結果顯示HMA誘導角質細胞終末分化能力較弱。 / 總而言之，本研究課題從兩種中藥中成功發現三個具有較強的抗銀屑病活性的化學單體成分，這些來自中藥的天然産物具有很好的開發成新的銀屑病治療外用制劑的應用前景。 / Psoriasis is an immunologically-mediated chronic inflammatory disease of the skin and joints affecting approximately 1-3% of the world’s population. Traditionally, Chinese medicine has been extensively used both inside and outside China for treating psoriasis with promising clinical results. Based on the promising findings in our previous screening project on 61 psoriasis-treating Chinese medicines which showed the root of Rubia cordifolia L. (Rubiae Radix) to have potent anti-psoriatic action, the present study aimed to identify active anti-psoriatic chemical constituents derived from Rubiae Radix and another Chinese herb namely Celastrus orbiculatus Thunb. and to elucidate the underlying mechanisms of action. / Microplate MTT assay was performed to evaluate the anti-proliferative actions of 28 selected Rubiae Radix-derived anthraquinones and other chemical ingredients on cultured HaCaT keratinocytes. Among them, 1-hydroxy-3-methyl-anthraquinone (HMA) and 1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone (DBA), as well as celastrol, a Celastrus orbiculatus-derived triterpene, were found to possess significant anti-proliferative action on HaCaT cells, with IC₅₀ value of 17.9, 15.8 and 1.1 μM, respectively. All DBA, HMA and celastrol showed only mild to moderate toxic effects on normal human keratinocyte HEK cells and human fibroblast Hs68 cells. / Mechanistically, celastrol and HMA was found to induce apoptosis in a dose-dependent manner in HaCaT cells as characterized by DNA fragmentation, phosphatidyl-serine externalization and activation of caspase 3. Further studies by flow cytometric and western blot analyses demonstrated that the celastrol-induced apoptosis on HaCaT cells was associated with the inhibition of NF-κB pathway and through caspase-related apoptotic pathway as characterized by activation of caspase proteins, regulation of Bcl-2 family proteins and depolarization of mitochondrial potential. / On the other hand, DBA showed an ability to induce terminal differentiation in cultured human keratinocytes and this capability is believed to be responsible for its growth inhibitory effects. DBA significantly accentuated the cornified envelope formation in HEK and HaCaT keratinocytes together with the augmentation of K1/K10, involucrin and transglutaminase 1 protein levels and decrease of expression of K5/K14 protein in DBA-treated cells. However, the subsequent in vivo study using a mouse tail model showed that HMA did not have significant effects on modulating keratinocyte terminal differentiation. / Taken together, our present PhD project successfully identified DBA, HMA and celastrol to have potent anti-psoriatic action on in vitro models, and the experimental findings render these naturally-occurring chemicals to be promising candidates for further development into anti-psoriatic pharmaceutical agents. / 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. / Zhou, Linli. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 213-244). / 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 (English) --- p.i / Abstract (Chinese) --- p.iii / Publications --- p.v / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Figures --- p.xvii / List of Tables --- p.xxi / List of Abbreviations --- p.xxii / Chapter Chapter One --- General Introduction / Chapter 1.1 --- Psoriasis --- p.2 / Chapter 1.1.1 --- Structure of skin --- p.2 / Chapter 1.1.2 --- Epidemiology of psoriasis --- p.3 / Chapter 1.1.3 --- Pathogenesis --- p.5 / Chapter 1.1.4 --- Classification --- p.8 / Chapter 1.1.4.1 --- Nonpustular (plaque type) psoriasis --- p.9 / Chapter 1.1.4.2 --- Guttate psoriasis --- p.9 / Chapter 1.1.4.3 --- Pustular psoriasis --- p.9 / Chapter 1.1.4.4 --- Erythrodermic psoriasis --- p.10 / Chapter 1.1.4.5 --- Nail psoriasis --- p.11 / Chapter 1.1.4.6 --- Psoriatic arthritis --- p.11 / Chapter 1.1.5 --- Comorbidities --- p.13 / Chapter 1.2 --- Treatment of Psoriasis --- p.16 / Chapter 1.2.1 --- Conventional treatment for psoriasis --- p.16 / Chapter 1.2.1.1 --- Topical therapy --- p.16 / Chapter 1.2.1.2 --- Phototherapy --- p.19 / Chapter 1.2.1.3 --- Systemic therapy --- p.21 / Chapter 1.2.2 --- Targeted immunotherapy --- p.24 / Chapter 1.2.3 --- Combination, rotational and sequential therapy --- p.25 / Chapter 1.2.4 --- Complementary treatment --- p.26 / Chapter 1.3 --- Traditional Chinese Medicine for Psoriasis --- p.30 / Chapter 1.3.1 --- Prescriptions for psoriasis based on pattern differentiation --- p.30 / Chapter 1.3.2 --- Clinical and experimental study of TCM for psoriasis --- p.34 / Chapter 1.3.3 --- Possible action mechanisms of Chinese herbs for psoriasis --- p.34 / Chapter 1.3.4 --- Previous studies on TCM for psoriasis conducted by our research group --- p.35 / Chapter 1.4 --- Aims and Objectives of the Present Study --- p.38 / Chapter Chapter Two --- Phytochemical and Apoptotic Studies of Rubiae Radix-derived Anthraquinones and Other Related Compounds / Chapter 2.1 --- Introduction --- p.41 / Chapter 2.2 --- Selection and Screening of Rubiae Radix-derived anthraquinones and Other Related Compounds for Anti-proliferative Action on Cultured HaCaT Human Keratinocytes --- p.43 / Chapter 2.2.1 --- Introduction --- p.43 / Chapter 2.2.2 --- Materials and methods --- p.45 / Chapter 2.2.2.1 --- Procurement of Rubiae Radix-derived anthraquiones and other related compounds --- p.45 / Chapter 2.2.2.2 --- Purification of anthraquinones from Rubiae Radix --- p.50 / Chapter 2.2.2.3 --- General cell culture --- p.54 / Chapter 2.2.2.4 --- SRB assay --- p.55 / Chapter 2.2.2.5 --- MTT assay --- p.56 / Chapter 2.2.2.6 --- Assessment of synergistic or antagonistic effects between two active anthraquiones --- p.56 / Chapter 2.2.2.7 --- Statistical analysis --- p.57 / Chapter 2.2.3 --- Results --- p.57 / Chapter 2.2.3.1 --- Anti-proliferative effects of 35 Rubiae Radix fractions on HaCaT cells by SRB assay --- p.57 / Chapter 2.2.3.2 --- Anti-proliferative effects of the 27 anthraquinones and related compounds on HaCaT cells by SRB assay --- p.59 / Chapter 2.2.3.3 --- Confirmation of the anti-proliferative action of 8 active pure compounds using MTT assay --- p.61 / Chapter 2.2.3.4 --- Cytotoxic effects of 1-hydroxy-3-methyl-anthraquinone and1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone on the growth of HEK and Hs68 cells --- p.64 / Chapter 2.2.3.5 --- Drug interactions between different active anthraquinones --- p.67 / Chapter 2.2.4 --- Discussion --- p.69 / Chapter 2.3 --- Investigations of the Apoptotic Effects of DBA and HMA on HaCaT cells --- p.71 / Chapter 2.3.1 --- Introduction --- p.71 / Chapter 2.3.2 --- Materials and methods --- p.76 / Chapter 2.3.2.1 --- Chemicals --- p.76 / Chapter 2.3.2.2 --- General cell culture methods --- p.76 / Chapter 2.3.2.3 --- Cell cycle analysis with PI staining --- p.76 / Chapter 2.3.2.4 --- Hoechst fluorescence staining for morphological evaluation --- p.77 / Chapter 2.3.2.5 --- DNA fragmentation assay --- p.77 / Chapter 2.3.2.6 --- Detection of apoptosis by flow cytometry --- p.78 / Chapter 2.3.2.7 --- Prepare cytosol fraction of HaCaT cells --- p.79 / Chapter 2.3.2.8 --- Western blot analysis --- p.79 / Chapter 2.3.2.9 --- Statistical analysis --- p.80 / Chapter 2.3.3 --- Results --- p.76 / Chapter 2.3.3.1 --- Action of DBA and HMA on cell cycle progression --- p.80 / Chapter 2.3.3.2 --- Alteration of cellular morphology --- p.84 / Chapter 2.3.3.3 --- Detection of DNA fragmentation --- p.86 / Chapter 2.3.3.4 --- Quantitative analysis of apoptotic cells by annexin V-PI staining --- p.88 / Chapter 2.3.3.5 --- Activation of procaspase-3 and release of cytochrome c protein --- p.91 / Chapter 2.3.4 --- Discussion --- p.94 / Chapter 2.4 --- General Discussion --- p.97 / Chapter Chapter Three --- Effects of Rubiae Radix and Its-derived Anthraquinones on Keratinocyte Terminal Differentiation / Chapter 3.1 --- Introduction --- p.100 / Chapter 3.2 --- Materials and Methods --- p.105 / Chapter 3.2.1 --- Chemicals --- p.105 / Chapter 3.2.2 --- General cell culture --- p.105 / Chapter 3.2.3 --- Cornified envelope (CE) formation assay --- p.106 / Chapter 3.2.4 --- Western blot analysis --- p.107 / Chapter 3.2.4 --- Statistical analysis --- p.107 / Chapter 3.3 --- Results --- p.108 / Chapter 3.3.1 --- EA fraction of Rubiae Radix, DBA and HMA stimulates CE formation --- p.108 / Chapter 3.3.2 --- EA fraction of Rubiae Radix, DBA and HMA regulated TG1 expression and involucrin production in cultured human keratinocytes --- p.112 / Chapter 3.3.3 --- Regulation of cytokeratins by EA fraction of Rubiae Radix, DBA and HMA --- p.118 / Chapter 3.4 --- Discussion --- p.128 / Chapter Chapter Four --- Anti-psoriatic Action of Celastrol from Celastrus orbiculatus / Chapter 4.1 --- Introduction --- p.136 / Chapter 4.2 --- Anti-proliferative Action of Celastrol on Cultured Human Keratinocytes and Other Cell Types --- p.138 / Chapter 4.2.1 --- Introduction --- p.138 / Chapter 4.2.2 --- Materials and methods / Chapter 4.2.2.1 --- Chemicals --- p.138 / Chapter 4.2.2.2 --- General cell culture --- p.139 / Chapter 4.2.2.3 --- MTT assay --- p.139 / Chapter 4.2.2.4 --- Statistical analysis --- p.139 / Chapter 4.2.3 --- Results --- p.142 / Chapter 4.2.3.1 --- Anti-proliferative effect of celastrol on cultured cells --- p.142 / Chapter 4.2.4 --- Discussion --- p.145 / Chapter 4.3 --- Induction of Apoptosis by Celastrol on Human Keratinocytes --- p.146 / Chapter 4.3.1 --- Introduction --- p.146 / Chapter 4.3.2 --- Materials and methods --- p.146 / Chapter 4.3.2.1 --- Chemicals --- p.146 / Chapter 4.3.2.2 --- General cell culture --- p.147 / Chapter 4.3.2.3 --- Cell cycle analysis with PI staining --- p.147 / Chapter 4.3.2.4 --- Detection of apoptosis by flow cytometry --- p.147 / Chapter 4.3.2.5 --- Measurement of the mitochondrial membrane potential (ΔΨm) --- p.148 / Chapter 4.3.2.6 --- Western blot analysis --- p.148 / Chapter 4.3.2.7 --- Statistical analysis --- p.148 / Chapter 4.3.3 --- Results --- p.149 / Chapter 4.3.3.1 --- Induction of sub-G1 phase by celastrol on HaCaT cells --- p.149 / Chapter 4.3.3.2 --- Quantitative analysis of apoptotic cells by Annexin V-PI staining --- p.151 / Chapter 4.3.3.3 --- Alteration of ΔΨm --- p.153 / Chapter 4.3.3.4 --- Activation of caspase family protein --- p.155 / Chapter 4.3.3.5 --- Celastrol regulates the Bcl-2 family members --- p.159 / Chapter 4.3.4 --- Discussion --- p.161 / Chapter 4.4 --- Inhibition of NF-κB Transcription Factor Activation by Celastrol --- p.164 / Chapter 4.4.1 --- Introduction --- p.164 / Chapter 4.4.2 --- Materials and methods --- p.165 / Chapter 4.4.2.1 --- Chemicals --- p.165 / Chapter 4.4.2.2 --- General cell cultrue --- p.165 / Chapter 4.4.2.3 --- Western blot analysis --- p.165 / Chapter 4.4.2.4 --- Detect nuclear p65 by ELISA assay --- p.166 / Chapter 4.4.2.5 --- Statistical analysis --- p.166 / Chapter 4.4.3 --- Results --- p.167 / Chapter 4.4.3.1 --- Celastrol inhibited the NF-κB activation --- p.167 / Chapter 4.4.4 --- Discussion --- p.170 / Chapter 4.5 --- Induction of Terminal Differentiation by Celastrol --- p.173 / Chapter 4.5.1 --- Introduction --- p.173 / Chapter 4.5.2 --- Materials and methods --- p.174 / Chapter 4.5.2.1 --- Chemicals --- p.174 / Chapter 4.5.2.2 --- General cell culture --- p.174 / Chapter 4.5.2.3 --- CE formation assay --- p.174 / Chapter 4.5.2.4 --- Western blot analysis --- p.174 / Chapter 4.5.2.5 --- Statistical analysis --- p.174 / Chapter 4.5.3 --- Results --- p.175 / Chapter 4.5.3.1 --- Regulation of CE formation by celastrol --- p.175 / Chapter 4.5.3.2 --- Modulation of terminal differentiation markers by celastrol --- p.178 / Chapter 4.5.4 --- Discussion --- p.181 / Chapter 4.6 --- General Discussion --- p.183 / Chapter Chapter Five --- In vivo Anti-psoriatic Effects of Topical Preparation of 1-hydroxy-3-methyl-anthraquinone / Chapter 5.1 --- Introduction --- p.187 / Chapter 5.2 --- Material and Methods --- p.191 / Chapter 5.2.1 --- Chemicals --- p.191 / Chapter 5.2.2 --- Formulation of topical preparation containing HMA --- p.191 / Chapter 5.2.3 --- Mouse tail model --- p.192 / Chapter 5.2.4 --- Histopathological evaluation --- p.193 / Chapter 5.2.5 --- Statistical analysis --- p.194 / Chapter 5.3 --- Results --- p.195 / Chapter 5.3.1 --- Body weight profile --- p.195 / Chapter 5.3.2 --- Histological resutls --- p.197 / Chapter 5.4 --- Discussion --- p.201 / Chapter Chapter Six --- General Conclusions and Future Perspectives / Chapter 6.1 --- General Conclusions --- p.205 / Chapter 6.2 --- Future Perspectives --- p.210 / References / References by alphabetical order --- p.213

銀屑病是一種慢性炎症性皮膚病，其發病率約佔全球1-3%的人口。銀屑病的病理特徵包括角質細胞增殖和分化異常，同時伴隨炎症反應，白細胞聚集於真皮和表皮以及血管擴張。證據顯示角質細胞能參與及延續免疫反應，以達致維持或促進該病的作用。研究亦建議角質細胞減少凋亡是引致銀屑病的一個特定現象；因此，長期以來誘導角質細胞凋亡就被用作為治療銀屑病的一種有效策略。 / 根據銀屑病的嚴重程度，治療方法可分為三級：外用藥物主要用於比較輕微的病患，而光療適合中等程度的病患；對於嚴重病例則可使用系統性治療或生物製劑。基於大約75%的銀屑病患者屬於輕微至中度病患，外用藥物是目前應用最為廣泛的治療方法。在中國銀屑病治療的歷史中曾經使用過中草藥，研究亦表明，其治療機制可能通過抑制角質細胞增殖和誘導角質細胞凋亡。比較研究也指出，傳統中藥比西藥的副作用相對較少，及具有較長的舒緩期和較低的復發率。 / 我們先前的研究發現，茜草根提取物能夠抑制一個和銀屑病相關的HaCaT角質細胞增殖。本研究證實，茜草根的乙酸乙酯提取物（EA）能誘導HaCaT細胞凋亡，其抑制角質細胞增殖的作用比茜草根的乙醇提取物（EE）更為有效，並可和一個流行於歐洲國家的重要外用銀屑病治療藥地蒽酚相比。另外，透過不同的檢測，包括形態學觀察，細胞凋亡雙染（磷脂結合蛋白V-碘化丙啶）分析，細胞週期分析，去氧核醣核酸斷裂測試，原位末端轉移酶標記技術，免疫熒光染色以及西方墨點法，我們發現一種在茜草中的化合物，1,4-二羥基-2-萘甲酸（DHNA）能通過死亡受體介導，線粒體介導或不依賴胱天蛋白酶的途徑導致HaCaT細胞凋亡。同時，在其中一種銀屑病動物模型，小鼠鼠尾鱗片表皮上的初步研究顯示DHNA亦可誘導角質細胞分化。此外，在細胞水平（存活率，釋放白细胞介素-1α）和動物上（Draize動物皮膚刺激性試驗）的實驗結果表明DHNA比地蒽酚的刺激性較小。 / 總括而言，本研究透過人類皮膚細胞和動物實驗說明EA和DHNA的細胞凋亡機制，以及DHNA對皮膚的潛在刺激性。這些結果顯示EA和DHNA有潛能發展成為安全及能有效治療銀屑病的替代藥物。EA和DHNA可在一個連續療程中結合使用，其中EA藥效媲美地蒽酚，應能迅速清除銀屑病皮損；而DHNA比地蒽酚的刺激性小，則比較適合應用在這個連續療程中後來的維護保養階段 / Psoriasis is a chronic inflammatory skin disorder that affects approximately 1-3% of the population worldwide. It is characterized by epidermal hyperplasia or abnormal differentiation, infiltration of leucocytes into the dermis and epidermis, dilation of blood vessels in dermis and inflammation. Evidence indicates keratinocytes contributed to the disease, and keratinocytes also participate in maintaining the chronically perpetuating immune response that sustains psoriasis. Decrease in keratinocytes apoptosis is suggested to be a specific pathogenic phenomenon, and induction of keratinocytes apoptosis have long been considered as an effective anti-psoriatic strategy. / Treatment of psoriasis is based on disease severity. Topical agents are predominantly for mild conditions; phototherapy for moderate conditions and systemic treatment or biological agents for severe cases. Topical treatment remains the most widely used method as an estimated 75% of psoriatic patients have mild to moderate disease. Chinese herbs have been used for the treatment of psoriasis in China, and studies showed their mechanism on treating psoriasis may through inhibition of keratinocyte proliferation and induction of apoptosis. Comparison studies also show that traditional Chinese medicine has relatively fewer side effects than western therapeutic agents, with a longer remission time and lower recurrence rate. / The extract of the root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) was previously found to inhibit keratinocyte proliferation using a psoriasis-relevant HaCaT cells model. In this study, the ethyl acetate extract of the root of Rubia cordifolia L. (EA) was confirmed to induce apoptosis on HaCaT cell, and the antiproliferative effect of EA is more potent than the ethanol extract of the herb (EE) and is comparable to dithranol, an important and popular topical treatment for psoriasis among Europe countries. Besides, we identified one of the components in Rubia cordifolia L., 1,4-dihydroxy-2-naphthoic acid (DHNA), could induce HaCaT keratinocyte apoptosis through the death receptor and mitochondria mediated pathway as well as in a caspase independent manner using various assays such as morphological examination, annexin V-PI staining, cell cycle analysis, DNA fragmentation, TUNEL assay, immunofluorescence staining and Western blot analysis. Moreover, DHNA was found to induce keratinocyte differentiation in a preliminary study using the in vivo mouse tail model of psoriasis. Furthermore, results from in vitro (cell viability, IL-1α release) and in vivo (Draize animal skin irritation test) experiments suggested DHNA have less irritation problems than dithranol. / In summary, this study describes the apoptotic mechanism of EA and DHNA, as well as the irritation potential of DHNA using different human skin cells and animal model. These results suggest EA and DHNA have the potential to develop as safe and effective therapeutic alternative for the treatment of psoriasis. EA and DHNA can be used together in a sequential therapy, in which EA is effective in rapid clearing of psoriatic lesions as its potency is comparable to dithranol; whereas DHNA is better suited for the later maintenance therapy for its milder irritation effect compared with dithranol. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Mok, Chong Fai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 164-183). / 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 (English version) --- p.iv / Abstract (Chinese version) --- p.vi / List of Publication and Presentation --- p.viii / Acknowledgements --- p.ix / Table of Contents --- p.x / List of Tables --- p.xvi / List of Figures --- p.xvii / List of Abbreviations --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Psoriasis --- p.1 / Chapter 1.1.1. --- Histological features --- p.2 / Chapter 1.1.2. --- Role of keratinocytes in psoriasis --- p.4 / Chapter 1.1.3. --- Decrease in skin cell apoptosis --- p.8 / Chapter 1.2. --- Treatment of psoriasis --- p.9 / Chapter 1.2.1. --- Conventional treatment --- p.9 / Chapter 1.2.1.1. --- Mild disease --- p.10 / Chapter 1.2.1.1.1. --- Corticosteroids --- p.10 / Chapter 1.2.1.1.2. --- Vitamin D₃ analogs --- p.11 / Chapter 1.2.1.1.3. --- Tazarotene --- p.11 / Chapter 1.2.1.1.4. --- Anthralin --- p.12 / Chapter 1.2.1.1.5. --- Coal tar --- p.12 / Chapter 1.2.1.2. --- Moderate disease --- p.12 / Chapter 1.2.1.2.1. --- Phototherapy --- p.12 / Chapter 1.2.1.3. --- Severe disease --- p.13 / Chapter 1.2.1.3.1. --- Retinoids --- p.13 / Chapter 1.2.1.3.2. --- Methotrexate --- p.14 / Chapter 1.2.1.3.3. --- Cyclosporine --- p.14 / Chapter 1.2.1.3.4. --- Fumaric acid --- p.15 / Chapter 1.2.1.3.5. --- Biological agents --- p.15 / Chapter 1.2.2. --- Alternative treatment --- p.16 / Chapter 1.2.2.1. --- Traditional Chinese Medicine (TCM) --- p.17 / Chapter 1.3. --- Aims and objectives of the present study --- p.19 / Chapter Chapter 2: --- Apoptotic Action of Ethyl Acetate Fraction of the Root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) on HaCaT Human Keratinocytes --- p.21 / Chapter 2.1. --- Introduction --- p.21 / Chapter 2.1.1. --- Rubia cordifolia L. --- p.21 / Chapter 2.1.2. --- Apoptosis --- p.22 / Chapter 2.1.3. --- Study objectives --- p.28 / Chapter 2.2. --- Materials and Methods --- p.30 / Chapter 2.2.1. --- Sources of medicinal materials --- p.30 / Chapter 2.2.2. --- Preparation of extracts --- p.30 / Chapter 2.2.3. --- Reagents --- p.31 / Chapter 2.2.4. --- Cell culture --- p.31 / Chapter 2.2.5. --- Proliferation assay --- p.32 / Chapter 2.2.6. --- Fluorescent staining for morphological evaluation --- p.33 / Chapter 2.2.7. --- Annexin V/propidium iodide staining --- p.33 / Chapter 2.2.8. --- JC-1 staining --- p.34 / Chapter 2.2.9. --- Statistical analysis --- p.35 / Chapter 2.3. --- Results --- p.36 / Chapter 2.3.1. --- EA inhibits proliferation of human epidermal HaCaT keratinocytes --- p.36 / Chapter 2.3.2. --- Alteration of cellular morphology --- p.39 / Chapter 2.3.3. --- EA increases phosphatidylserine externalization in HaCaT cells --- p.41 / Chapter 2.3.4. --- EA decreases MMP --- p.45 / Chapter 2.4. --- Discussion --- p.47 / Chapter Chapter 3: --- Identification of Pure Compound for Possible Apoptotic Action on HaCaT Human Keratinocytes and Detailed Mechanistic Study --- p.51 / Chapter 3.1. --- Introduction --- p.51 / Chapter 3.1.1. --- Anthraquinone --- p.51 / Chapter 3.1.2. --- Study objectives --- p.52 / Chapter 3.2. --- Materials and Methods --- p.54 / Chapter 3.2.1. --- Reagents --- p.54 / Chapter 3.2.2. --- Cell culture --- p.54 / Chapter 3.2.3. --- Proliferation assay --- p.55 / Chapter 3.2.4. --- Fluorescent staining for morphological evaluation --- p.56 / Chapter 3.2.5. --- Annexin V/propidium iodide staining --- p.56 / Chapter 3.2.6. --- JC-1 staining --- p.56 / Chapter 3.2.7. --- Cell cycle analysis --- p.56 / Chapter 3.2.8. --- Detection of DNA fragmentation --- p.57 / Chapter 3.2.9. --- Terminal Deoxynucleotidyltransferase-Mediated dUTP Nick End Labeling (TUNEL) assay --- p.57 / Chapter 3.2.10. --- Western blot analysis --- p.58 / Chapter 3.2.11. --- Immunofluorescence staining --- p.59 / Chapter 3.2.12. --- Statistical analysis --- p.60 / Chapter 3.3. --- Results --- p.61 / Chapter 3.3.1. --- DHNA inhibits proliferation of human epidermal HaCaT Keratinocytes --- p.61 / Chapter 3.3.2. --- Alteration of cellular morphology --- p.70 / Chapter 3.3.3. --- DHNA increases phosphatidylserine externalization in HaCaT cells --- p.72 / Chapter 3.3.4. --- DHNA decreases MMP --- p.76 / Chapter 3.3.5. --- DHNA causes G0/G1 cell cycle arrest in HaCaT cells --- p.78 / Chapter 3.3.6. --- DHNA increases DNA fragmentation --- p.81 / Chapter 3.3.7. --- DHNA increases TUNEL positive cells in HaCaT cells --- p.83 / Chapter 3.3.8. --- Western blot analysis --- p.85 / Chapter 3.3.9. --- DHNA induced Fas aggregation in HaCaT cells --- p.88 / Chapter 3.3.10. --- Caspase inhibition assay --- p.90 / Chapter 3.3.11. --- DHNA induced caspase independent apoptosis in HaCaT cells --- p.93 / Chapter 3.3.12. --- Effects of DHNA on MAPK in HaCaT cells --- p.96 / Chapter 3.3.13. --- MAPK inhibition assay --- p.100 / Chapter 3.4. --- Discussion --- p.104 / Chapter Chapter 4: --- Anti-Psoriatic Effects of Topical 1,4-Dihydroxy-2-naphthoic acid Formulation on in vivo Mouse Tail Experiments --- p.111 / Chapter 4.1. --- Introduction --- p.111 / Chapter 4.1.1. --- Keratinocytes differentiation process --- p.111 / Chapter 4.1.2. --- Animal model for psoriasis --- p.114 / Chapter 4.1.3. --- Study objectives --- p.119 / Chapter 4.2. --- Materials and Methods --- p.122 / Chapter 4.2.1. --- Reagents --- p.122 / Chapter 4.2.2. --- Formulation and preparation of topical drug --- p.122 / Chapter 4.2.3. --- Mice for in vivo experiments --- p.123 / Chapter 4.2.4. --- Treatment with topical preparations --- p.124 / Chapter 4.2.5. --- Statistical analysis --- p.125 / Chapter 4.3. --- Results --- p.126 / Chapter 4.3.1. --- Tail skin appearance after topical treatment --- p.126 / Chapter 4.3.2. --- Histological examination and findings --- p.128 / Chapter 4.4. --- Discussion --- p.132 / Chapter Chapter 5: --- Prediction of Skin Irritation Potential of 1,4-Dihydroxy-2-naphthoic acid by in vitro and in vivo Experiments --- p.135 / Chapter 5.1. --- Introduction --- p.135 / Chapter 5.1.1. --- Skin irritation --- p.135 / Chapter 5.1.2. --- Viability test and IL-1α release --- p.136 / Chapter 5.1.3. --- Animal irritation test --- p.139 / Chapter 5.1.4. --- Study objectives --- p.139 / Chapter 5.2. --- Materials and Methods --- p.141 / Chapter 5.2.1. --- Reagents --- p.141 / Chapter 5.2.2. --- Cell culture --- p.141 / Chapter 5.2.3. --- Viability test --- p.141 / Chapter 5.2.4. --- IL-1α release assay --- p.142 / Chapter 5.2.5. --- Animal irritation test --- p.142 / Chapter 5.2.6. --- Statistical analysis --- p.143 / Chapter 5.3. --- Results --- p.144 / Chapter 5.3.1. --- Viability test --- p.144 / Chapter 5.3.2. --- IL-1α release assay --- p.144 / Chapter 5.3.3. --- Animal irritation test --- p.147 / Chapter 5.4. --- Discussion --- p.152 / Chapter Chapter 6: --- General Discussion and Conclusions --- p.155 / References --- p.164

Chan, Yuet Wa. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 217-232). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via 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

Li Mi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 125-136). / Abstracts in English and Chinese. / Abstract --- p.i / 論文摘要 --- p.iv / Publications --- p.vi / Acknowledgement --- p.vii / Abbreviations --- p.viii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Pyrrolizidine alkaloids --- p.1 / Chapter 1.1.1 --- Distribution and plant sources --- p.1 / Chapter 1.1.2 --- Structures and nomenclature --- p.3 / Chapter 1.2 --- PA-containing Chinese medicinal herbs --- p.6 / Chapter 1.3 --- PA-induced toxicity。 --- p.7 / Chapter 1.3.1 --- Acute toxicity and chronic toxicity --- p.7 / Chapter 1.3.2 --- Genotoxicity --- p.8 / Chapter 1.3.3 --- Tumorigenicity --- p.8 / Chapter 1.3.4 --- Hepatotoxicity --- p.8 / Chapter 1.3.5 --- Mechanism of toxic effects --- p.9 / Chapter 1.3.5.1 --- Metabolic pathways --- p.10 / Chapter 1.3.5.2 --- Liver tissue-bound pyrroles --- p.16 / Chapter 1.3.5.3 --- Metabolizing enzymes --- p.17 / Chapter 1.3.5.3.1 --- Phase I metabolizing enzymes --- p.17 / Chapter 1.3.5.3.2 --- Phase II metabolizing enzymes --- p.18 / Chapter 1.3.5.4 --- Species and gender specificity toward toxicity --- p.19 / Chapter 1.3.5.5 --- Structure-activity relationships --- p.20 / Chapter 1.4 --- Prevention of PAs-induced toxicity --- p.23 / Chapter 1.4.1 --- Significance of prevention in humans --- p.23 / Chapter 1.4.2 --- Regulations toward preventing toxicity induced by PAs --- p.24 / Chapter 1.5 --- Aim of the present study --- p.25 / Chapter Chapter 2 --- Qualitative and Quantitative Analysis of PA-containing Chinese Medicinal Herbs --- p.26 / Chapter 2.1 --- Materials and equipments --- p.27 / Chapter 2.1.1 --- Herbal materials --- p.27 / Chapter 2.1.2 --- Chemicals and solvents --- p.27 / Chapter 2.1.3 --- Equipment and instrumentation --- p.27 / Chapter 2.2 --- Preparation of herbal extracts。 --- p.28 / Chapter 2.2.1 --- Crude herbal extract --- p.28 / Chapter 2.2.2 --- Total pyrrolizidine alkaloid extract --- p.28 / Chapter 2.3 --- Qualitative and quantitative analysis of Ligularia hodgsonii --- p.29 / Chapter 2.3.1 --- Methods --- p.29 / Chapter 2.3.1.1 --- HPLC-UV condition --- p.29 / Chapter 2.3.1.2 --- HPLC-MS condition --- p.29 / Chapter 2.3.1.3 --- Calibration curve for clivorine --- p.29 / Chapter 2.3.1.4 --- Recovery test --- p.30 / Chapter 2.3.1.5 --- Sample test --- p.30 / Chapter 2.3.2 --- Results and discussions --- p.30 / Chapter 2.3.2.1 --- Qualitative analysis of PAs in Ligularia hodgsonii --- p.30 / Chapter 2.3.2.2 --- Calibration curve for clivorine --- p.35 / Chapter 2.3.2.3 --- Result of the recovery test --- p.37 / Chapter 2.3.2.4 --- Quantification of PAs in Ligularia hodgsonii --- p.37 / Chapter 2.4 --- Qualitative and quantitative analysis of Tussilago farfara --- p.37 / Chapter 2.4.1 --- Methods --- p.38 / Chapter 2.4.1.1 --- HPLC-UV condition --- p.38 / Chapter 2.4.1.2 --- HPLC-MS condition --- p.39 / Chapter 2.4.1.3 --- Calibration curve for senkirkine --- p.39 / Chapter 2.4.1.4 --- Recovery test --- p.39 / Chapter 2.4.1.5 --- Sample test --- p.39 / Chapter 2.4.2 --- Results and discussions --- p.40 / Chapter 2.4.2.1 --- Qualitative analysis of senkirkine in Tussilago farfara --- p.40 / Chapter 2.4.2.2 --- Calibration curve for senkirkine --- p.42 / Chapter 2.4.2.3 --- Result of the recovery test --- p.42 / Chapter 2.4.2.4 --- Quantification of senkirkine in Tussilago farfara --- p.43 / Chapter 2.5 --- Qualitative and quantitative analysis of Gynura segetum --- p.43 / Chapter 2.5.1 --- Methods --- p.43 / Chapter 2.5.1.1 --- HPLC-UV condition --- p.43 / Chapter 2.5.1.2 --- HPLC-MS condition --- p.44 / Chapter 2.5.1.3 --- Calibration curves for senecionine and seneciphylline --- p.44 / Chapter 2.5.1.4 --- Recovery test… --- p.44 / Chapter 2.5.1.5 --- Sample test --- p.44 / Chapter 2.5.2 --- Results and discussions。 --- p.45 / Chapter 2.5.2.1 --- Qualitative analysis of PAs in Gynura segetum --- p.45 / Chapter 2.5.2.2 --- Calibration curves for senecionine and seneciphylline --- p.49 / Chapter 2.5.2.3 --- Result of the recovery test --- p.49 / Chapter 2.5.2.4 --- Quantification of PAs in Gynura segetum --- p.49 / Chapter 2.6 --- Qualitative and quantitative analysis of Crotalaria sessiliflora --- p.50 / Chapter 2.6.1 --- Methods --- p.50 / Chapter 2.6.1.1 --- HPLC-UV condition --- p.50 / Chapter 2.6.1.2 --- HPLC-MS condition --- p.50 / Chapter 2.6.1.3 --- Calibration curve --- p.51 / Chapter 2.6.1.4 --- Recovery test --- p.51 / Chapter 2.6.1.5 --- Sample test --- p.51 / Chapter 2.6.2 --- Results and discussions --- p.52 / Chapter 2.6.2.1 --- Qualitative analysis of monocrotaline in Crotalaria sessiliflora --- p.52 / Chapter 2.6.2.2 --- Calibration curve for monocrotaline --- p.54 / Chapter 2.6.2.3 --- Result of the recovery test --- p.54 / Chapter 2.6.2.4 --- Quantification of PAs in Crotalaria sessiliflora --- p.55 / Chapter 2.7 --- Qualitative and quantitative analysis of Senecio scandens --- p.55 / Chapter 2.7.1 --- Methods --- p.55 / Chapter 2.7.1.1 --- HPLC-UV condition --- p.55 / Chapter 2.7.1.2 --- HPLC-MS condition --- p.56 / Chapter 2.7.1.3 --- Sample test --- p.56 / Chapter 2.7.2 --- Results and discussions --- p.56 / Chapter 2.7.2.1 --- Qualitative analysis of PAs in Senecio scandens --- p.56 / Chapter 2.7.2.2 --- Quantification of PAs in Senecio scandens --- p.59 / Chapter Chapter 3 --- Hepatotoxicity Induced by PA-containing Chinese Medicinal Herbs --- p.60 / Chapter 3.1 --- Materials and methods --- p.62 / Chapter 3.1.1 --- Reagents --- p.62 / Chapter 3.1.2 --- Animal models --- p.62 / Chapter 3.1.3 --- Determination of the serum ALT activity --- p.64 / Chapter 3.1.4 --- Determination of hepatic GSH level --- p.68 / Chapter 3.1.5 --- Quantitation of liver tissue-bound pyrroles --- p.69 / Chapter 3.1.6 --- Histological assessment of liver morphological changes --- p.70 / Chapter 3.1.7 --- Assessment of hepatocytes apoptosis --- p.71 / Chapter 3.1.8 --- Statistical analysis --- p.72 / Chapter 3.2 --- Results and discussion --- p.72 / Chapter 3.2.1 --- Calibration curves --- p.72 / Chapter 3.2.1.1 --- Calibration curve for the determination of serum ALT activity --- p.72 / Chapter 3.2.1.2 --- Calibration curve of determination of hepatic GSH level --- p.73 / Chapter 3.2.2 --- Hepatotoxicity Study of Crotalaria sessiliflora --- p.74 / Chapter 3.2.2.1 --- Hepatotoxicity at 24 hrs after treatment --- p.74 / Chapter 3.2.2.1.1 --- Correlation between dosage of monocrotaline in Crotalaria sessiliflora and amount of liver tissue-bound pyrroles --- p.74 / Chapter 3.2.2.1.2 --- Effects of Crotalaria sessiliflora on the serum ALT activity --- p.79 / Chapter 3.2.2.1.3 --- The correlation between the elevated level of ALT activity and apoptosis of liver cells --- p.85 / Chapter 3.2.2.1.4 --- Effects of Crotalaria sessiliflora on the hepatic GSH level --- p.86 / Chapter 3.2.2.1.5 --- Histological changes of liver sections --- p.89 / Chapter 3.2.2.2 --- Hepatotoxicity within 4 days after administration --- p.92 / Chapter 3.2.2.3 --- Sub-acute hepatotoxicity within 14 days after administration --- p.93 / Chapter 3.2.2.4 --- Conclusion in hepatotoxicity study of Crotalaria sessiliflora --- p.99 / Chapter 3.2.3 --- Hepatotoxicity Study of Gynura segetum --- p.102 / Chapter 3.2.3.1 --- Correlation between the dosage of PAs present in Gynura segetum and the amount of liver tissue-bound pyrroles --- p.102 / Chapter 3.2.3.2 --- "Effects of Gynura segetum on serum ALT activity, hepatic GSH level and morphological changes of liver" --- p.104 / Chapter 3.2.4 --- Hepatotoxicity Study of Ligularia hodgsonii --- p.108 / Chapter 3.2.4.1 --- Correlation between the dosage of PAs present in Ligularia hodgsonii and the formation of liver tissue-bound pyrroles --- p.108 / Chapter 3.2.4.2 --- Effects of Ligularia hodgsonii on serum ALT activity and hepatic GSH level --- p.111 / Chapter 3.2.5 --- Hepatotoxicity Study of Tussilago farfara --- p.113 / Chapter 3.2.6 --- Hepatotoxicity Study of PA-containing medicinal herbs --- p.115 / Chapter 3.2.6.1 --- Correlation between formation of liver tissue-bound pyrroles and elevated serum ALT level --- p.115 / Chapter 3.2.6.2 --- Correlation between dosage of PAs and amount of liver tissue-bound pyrroles --- p.117 / Chapter 3.2.7 --- Test of Liver Tissue-bound Pyrroles as a biomarker using Senecionis scandentis --- p.118 / Chapter 3.2.8 --- Conclusions --- p.120 / Chapter Chapter 4 --- General Conclusions --- p.121 / Chapter 4.1 --- Qualitative and quantitative analysis of five PA-containing medicinal herbs --- p.121 / Chapter 4.2 --- Hepatotoxicity induced by PA-containing medicinal herbs in rats --- p.122 / Chapter 4.3 --- The correlation between hepatotoxicity induced by PA-containing medicinal herbs and the formation of liver tissue-bound pyrroles --- p.123 / Chapter 4.4 --- Threshold of the amount of liver tissue-bound pyrroles related to the hepatotoxicity induced by PA-containing medicinal herbs --- p.123 / References --- p.125

Dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Master of Science in Medicine. Johannesburg 2012. / Dental caries and periodontal diseases are the most common bacterial oral infections in humans. Prevention and treatment of these infections are oral hygiene measures including antimicrobial compounds which can be chemical agents or natural products. Dodonaea viscosa var. angustifolia is an indigenous South African plant that has antibacterial, antiviral and anticandida properties. This study investigated the antibacterial and antivirulence effect of D. viscosa var. angustifolia on cariogenic bacteria and the causative organisms of periodontal diseases.

Halenia elliptica D. Don belongs to Gentianaceae family. It is often used as part of a traditional Tibetan medicine to treat hepatitis. In the present investigation, six major xanthone components were isolated and identified from Halenia elliptica. An HPLC/DAD/APCI/MS method was developed and validated for the quantitative analysis of these xanthones, including 1-hydroxy-2,3,5-trimethoxy-xanthone (HM-1), 1-hydroxy-2,3,4,7-tetramethoxy- xanthone (HM-2), 1-hydroxy-2,3,4,5-tetramethoxy-xanthone (HM-3), 1,7- dihydroxy-2,3,4,5-tetramethoxy-xanthone (HM-4), 1,5-dihydroxy-2,3-dimethoxy-xanthone (HM-5) and 1-0-[beta-D-xylopyranosyl-(1-6)-beta-D-glucopyranosyl]-2,3,5-trimethoxy-xanthone (HM-2-10). All the xanthones aglycons caused vasodilation in the coronary artery pre-contracted with 1 muM 5-HT, but the xanthone glycoside had no effect. HM-1 was one of the most abundant xanthones with the most potent vasorelaxant activity. / Mechanisms of the vasorelaxant effect of HM-1 were investigated. HM-1 showed a potent vasorelaxant activity on rat coronary artery involved both an endothelium-dependent mechanism involving NO and an endothelium-independent mechanism by inhibiting Ca2+ influx through L-type voltage-operated Ca2+ channels. / Taken together, in spite of the pharmacokinetics results showed that HM-1 was rapidly and widely distributed to tissues after intravenous administration in rats, with conjugation to being the major metabolic pathway in vivo, both HM-1 and its active metabolite (HM-5) show that they are important pharmacological agents with potentially useful therapeutic indications. / The metabolism and pharmacokinetics of HM-1 displayed biphasic elimination kinetics, with an elimination half-life of 60.4 +/- 4.2 min. Four other Phase I metabolites were isolated and identified as demethylated products in vitro. HM-1 was metabolised to HM-5 in the liver. Biliary excretion studies showed that both HM-1 and the metabolite (HM-5) underwent extensive phase II conjugation to form glucuronides and sulfates. Tissue distribution studies showed that HM-1 was widely distributed to different organs. Collection of urine and faeces over 24 h showed that 10.88% of dose was excreted from urine and 1.91% of dose via faeces. / With HM-5 being one of the major in vivo metabolites of HM-1, the effect of HM-5 has been studied on rat coronary artery and compared to HM-1. HM-5-mediated vasorelaxant effect was mediated through opening of potassium channel (TEA, 4-AP) and altering intracellular calcium by partial inhibition of Ca2+ influx through L-type voltage-operated Ca 2+ channels and intracellular Ca2+ stores. / "September 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4699. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 195-218). / 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.

by Cheng Yuk Kwan, Anna. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 104-112). / Acknowledgements / Table of Contents --- p.i / Abbreviations --- p.iv / Abstract --- p.vi / List of figures --- p.ix / List of tables --- p.xi / Chapter Chapter One: --- Introduction / Chapter 1.1 --- Objective and scope of the project --- p.1 / Chapter 1.2 --- Literature review of Mimosa pudica / Chapter 1.2.1 --- Morphology of Mimosa pudica --- p.3 / Chapter 1.2.2 --- Chemistry of Mimosa pudica --- p.5 / Chapter 1.2.3 --- Uses in traditional medicine --- p.5 / Chapter 1.2.4 --- Clinical and pharmacological studies of Mimosa pudica --- p.6 / Chapter 1.2.5 --- Toxicology of Mimosa pudica --- p.8 / Chapter 1.2.6 --- Characteristics and toxicology of mimosine --- p.9 / Chapter 1.3 --- Immunomodulation / Chapter 1.3.1 --- Overview of the immune system --- p.11 / Chapter 1.3.2 --- Strategies on the study of immunomodulation of Mimosa pudica --- p.13 / Chapter 1.4 --- Toxicology / Chapter 1.4.1 --- Principles of the toxicological assays / Chapter 1.4.1.1 --- LD50 --- p.17 / Chapter 1.4.1.2 --- Enzyme assays --- p.18 / Chapter 1.4.1.3 --- Subacute toxicity test --- p.24 / Chapter 1.4.1.4 --- Reproductive toxicity test --- p.25 / Chapter Chapter Two: --- Materials and methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Mimosa pudica --- p.27 / Chapter 2.1.2 --- Animals --- p.27 / Chapter 2.1.3 --- Chemicals --- p.28 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Extraction of Mimosa pudica --- p.32 / Chapter 2.2.2 --- Assays for the immunomodulatory effects of Mimosa pudica / Chapter 2.2.2.1 --- Cell preparation / Chapter a) --- Splenocytes --- p.35 / Chapter b) --- Thymocytes --- p.35 / Chapter c) --- Macrophages --- p.36 / Chapter 2.2.2.2 --- Splenocyte proliferation --- p.37 / Chapter 2.2.2.3 --- Thymocyte proliferation --- p.38 / Chapter 2.2.2.4 --- Phagocytic activity of macrophages --- p.39 / Chapter 2.2.2.5 --- Release of IL-1 by macrophages --- p.40 / Chapter 2.2.2.6 --- Plaque forming cells --- p.41 / Chapter 2.2.2.7 --- Restoration on splenocyte blastogenesis of old mice --- p.42 / Chapter 2.2.3 --- Assays for the toxicity of Mimosa pudica / Chapter 2.2.3.1 --- LD50 --- p.43 / Chapter 2.2.3.2 --- Enzyme assays --- p.43 / Chapter 2.2.3.3 --- Subacute toxicity --- p.43 / Chapter 2.2.3.4 --- Reproductive toxicity --- p.44 / Chapter 2.2.4 --- Statistical analysis --- p.44 / Chapter Chapter Three: --- Results / Chapter 3.1 --- Immunomodulatory effects of Mimosa pudica / Chapter 3.1.1 --- In vitro study on the lymphocyte proliferation / Chapter 3.1.1.1 --- Splenocyte proliferation --- p.45 / Chapter 3.1.1.2 --- Thymocyte proliferation --- p.50 / Chapter 3.1.2 --- In vivo study on the lymphocyte proliferation --- p.53 / Chapter 3.1.3 --- Phagocytic activity of macrophages --- p.58 / Chapter 3.1.4 --- Release of IL-1 by macrophages --- p.64 / Chapter 3.1.5 --- Plaque forming cells --- p.67 / Chapter 3.1.6 --- Restoration on splenocyte blastogenesis of old mice --- p.69 / Chapter 3.2 --- Toxicity of Mimosa pudica / Chapter 3.2.1 --- LD50 --- p.72 / Chapter 3.2.2 --- Enzyme assays --- p.75 / Chapter 3.2.3 --- Subacute toxicity --- p.80 / Chapter 3.2.4 --- Reproductive toxicity --- p.85 / Chapter Chapter Four: --- General discussion on the immunomodulatory effects and toxicity of Mimosa pudica / Chapter 4.1 --- Immunomodulatory effects of Mimosa pudica --- p.88 / Chapter 4.2 --- Toxicity of Mimosa pudica --- p.95 / Chapter Chapter Five: --- Concluding remarks --- p.99 / References --- p.104 / Appendix --- p.113

Cheung, Wing Shing David. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 201-217). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Law, Wai Tak. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 107-123). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iv / Acknowledgements --- p.vi / Publications --- p.viii / Table of Contents --- p.ix / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Wound healing --- p.1 / Chapter 1.1.1 --- Physiology of wound healing --- p.1 / Chapter 1.1.2 --- Three phases of wound healing --- p.3 / Chapter 1.1.3 --- Angiogenesis in wound healing --- p.10 / Chapter 1.2 --- Delayed wound healing --- p.11 / Chapter 1.2.1 --- Chronic ulcers --- p.11 / Chapter 1.2.2 --- Examples of ulcers --- p.12 / Chapter 1.3 --- Traditional Chinese medicine (TCM) --- p.16 / Chapter 1.3.1 --- Principles of TCM --- p.16 / Chapter 1.3.2 --- TCM and chronic ulcers --- p.16 / Chapter 1.4 --- Objectives of study --- p.19 / Chapter Chapter 2 --- Materials and Methods --- p.21 / Chapter 2.1 --- Selection of traditional Chinese herbs --- p.21 / Chapter 2.2 --- Authentication of TCM --- p.22 / Chapter 2.3 --- Preparation of TCM --- p.23 / Chapter 2.4 --- In vitro studies on the effects of TCM on wound healing --- p.23 / Chapter 2.4.1 --- Angiogenesis study by using human umbilical vein endothelial cell (HUVEC) --- p.25 / Chapter 2.4.2 --- Granulation study by using human fibroblast cell line (CRL) --- p.32 / Chapter 2.4.3 --- Preparation of cell culture conditions --- p.35 / Chapter 2.5 --- In vivo study on the effects of TCM on wound healing by using diabetic mice --- p.38 / Chapter 2.5.1 --- Diabetic mice model --- p.38 / Chapter 2.5.2 --- Diabetic mice wound induction --- p.41 / Chapter 2.5.3 --- "Measurement of body weight, blood glucose level and ulcer area" --- p.43 / Chapter Chapter 3 --- Results / Chapter 3.1 --- The percentage yield of each herbs --- p.48 / Chapter 3.2 --- pH value of all the effective treatment concentration --- p.49 / Chapter 3.3 --- Selection of traditional Chinese herbs --- p.53 / Chapter 3.4 --- Effect of selected TCM on the proliferation of HUVEC --- p.55 / Chapter 3.5 --- Effect of selected TCM on the migration of HUVEC --- p.61 / Chapter 3.6 --- Effect of selected TCM on the proliferation of CRL --- p.63 / Chapter 3.7 --- "Effect of Radix Rehmanniae (selected TCM) on the change in body weight, blood glucose level and ulcer area" --- p.66 / Chapter Chapter 4 --- Discussions --- p.75 / Chapter Chapter 5 --- How does my study contribute towards the modernisation of Chinese medicine? --- p.100 / References --- p.107 / Appendix --- p.124