Thesis submitted in fulfilment of the requirements for the degree Magister Technologiae: Chemistry in the Faculty of Applied Sciences at the CAPE PENINSULA UNIVERSITY OF TECHNOLOGY 2014 / The solid state modification of a given active pharmaceutical ingredient is a desired way to alter its physicochemical properties, such as solubility or bioavailability. The solubilitymelting point relationship of the ensuing co-crystal or salt is not fully understood. In this thesis, a series of model co-crystals and pharmaceutical co-crystals and salts of baclofen were investigated. The model co-crystals were prepared from 4,4’-bipyridine (BIPY) and 1,2-bis(4-pyridyl)ethane (ETBIPY) used as host compounds which were combined with a series of carboxylic acids as co-formers, such as p-toluic acid (PTA), rac-phenylbutyric acid (racPBA), racemic and S-phenylsuccinic acid (racPSA and S-PSA, respectively). In the second part, six new multicomponent crystals of baclofen (BAC, (RS) 4-amino-3-(4- chlorophenyl)-butanoic acid), were prepared with mono- and dicarboxylic acids: two pharmaceutical co-crystals obtained with benzoic acid (BAC•BA) and p-toluic acid (BAC•PTA) and four pharmaceutical salts with 1-hydroxy-2-naphthoic acid, (BAC+)(HNA-), oxalic acid, 2(BAC+)(OA2-), maleic acid, (BAC+)(MA-) and p-toluene sulfonic acid, (BAC+)(PTSA-)•IPA. The compounds prepared were analysed by single crystal and powder X-ray diffractometry, differential scanning calorimetry and their solubility was measured in water and ethanol. From the analysis of the model co-crystals it was concluded that their aqueous solubility is inversely related to the melting point values and this can be explained by packing features. Also, the introduction of a chiral building block, compared to its racemic counterpart, is a valuable way to limit the formation of the intermolecular interactions in the new multicomponent crystal and thus decrease the efficiency of the packing which eventually leads to lower melting points and better solubility. The analysis of the baclofen crystals suggests that a strong, robust and predictable hydrogen bonding network with a combination of molecular building blocks which show acceptable molecular flexibility is a good recipe for successful co-crystal design.

by Wing-yu Tang. / Bibliography: leaves 129-138 / Thesis (M.Ph.)--Chinese University of Hong Kong, 1985

多年來，儘可能多的從植物中提取單體化合物一直是開發新型化學防癌劑和化學治療劑藥物的重要來源。 / 在本课题中，我们活性測試了从刺果紫玉盘（番荔枝科植物）中分离得到的14个番荔枝内酯化合物和7个多氧环己烯化合物，从三叉刺（豆科植物）和黄瑞香（瑞香科植物）中分离得到的4个黄酮化合物，从黄瑞香（瑞香科植物）和了哥王（瑞香科植物）中分离得到的2个香豆素化合物，以及从总状蕨藻（蕨藻科植物）中分离得到的1 个生物碱化合物，對11種人類常見癌症細胞株，如惡性黑色素瘤、肺癌、子宮頸上皮腺癌、肝癌、前列腺癌、結直腸癌的體外抗癌活性，用以建立一個全面的抗癌活性數據庫，為人們更好得了解番荔枝科植物奠定基礎。 / 在這些被篩選的單體化合物中，番荔枝內酯（ACGs）顯示出卓越的抗癌活性。它們對某些癌細胞株的細胞毒性甚至達到了nmol/l級別。例如番荔枝內酯desacetyluvaricin（Dau），對11條人類癌細胞株具有廣泛的抗增生活性，其半抑制濃度（IC₅₀）範圍從2.3 nM到37.4 μM。其中，Dau對結直腸癌細胞SW480的毒性最甚。Dau不僅具有高的抗癌效力，并對人正常纖維細胞Hs68幾乎沒有細胞毒性，半抑制濃度超過了247.5 μM。進一步的機理研究表明，Dau可導致SW480細胞產生大量過氧化物，進而導致細胞核內DNA斷裂。DNA損傷會讓MEK/ERK信號通路失活，並且影響了細胞週期調控蛋白的正常表達。例如影響細胞S週期的調控蛋白Cyclin A和Cyclin E的表達，以及影響G₁/S檢查點調控蛋白E2F的表達。由此，Dau促進SW480癌細胞穿過G₁/S檢查點，由G₁進入S期並在S期累計。最終被抑制在S週期的SW480細胞發生了壞死。以上機理的研究可為更好的理解ACG的作用機制提供一定的理論基礎。 / 番荔枝內酯是一系列長鏈脂肪酸的衍生物。它的結構的多樣性引發了我們極大的興趣去研究它的構效關係。我們比較了14個番荔枝內酯在細胞毒性和細胞週期控制方面對兩種不同的前列腺癌細胞LNCaP（p53基因野生型）和PC-3（p53基因缺失型）的影響。實驗結果表明，LNCaP細胞比PC-3更加敏感。番荔枝內酯的這種選擇性大概跟癌細胞中p53抑癌蛋白的表達水平有關。此外，關於構效關係的研究我們還發現：（1）在番荔枝內酯結構的核心系統中，四氫呋喃環的個數越多，化合物的抗癌活性越高；（2）在含有相鄰雙四氫呋喃環結構的化合物中，擁有threo/trans/threo/trans/erythro立體構型的化合物的細胞毒性比擁有threo/trans/threo/trans/threo立體構型的化合物高；（3）含單或雙四氫呋喃環結構的番荔枝內酯都將通過將LNCaP細胞抑制在G₁/G₀週期從而達到抗癌效果，並不會引起細胞凋亡；（4）含單四氫呋喃環結構的番荔枝內酯都將通過引發細胞凋亡從而達到抑制PC-3癌細胞的增長。然而含雙四氫呋喃環結構的番荔枝內酯會引發更多的PC-3細胞凋亡，並且有不同程度的細胞週期抑制；（5）在四氫呋喃環核心體系上，乙酰氧基會比羥基增加番荔枝內酯的細胞毒性；（6）雙鍵的取代基也會增加毒性效果。我們的研究結果印證了一些文獻已報導的關於番荔枝內酯構效關係的結論，同時我們也提出了一些新的假設。 / 本研究不僅增加了我們對番荔枝內酯強大的抗癌活性更全面的了解，並且通過機理研究還為它的選擇性毒性及構效關係特點提供了有重要的信息。番荔枝內酯是一類具有充滿前景抗癌化合物。在接下來的研究中，我們將致力於體內抗癌活性的研究，并擴大研究範圍，通過對多個ACG化合物的機理研究來證明我們對它的選擇性毒性的機理假設。 / For years and years, the discovery of phytochemicals as many as possible has always been an important strategy for the development of novel chemopreventive and chemotherapeutic drugs. / In this studies, we have screened 14 Annonaceous acetogenins and 7 polyoxygenated cyclohexenes isolated from the root of Uvaria calamistrata (Annonaceae), 4 flavonoids isolated from the stems of Trifidacanthus unifoliolatus (Fabaceae) and Daphne giraldii (Thymelaeaceae), 2 cumarins isolated from the stem bark of Daphne giraldii (Thymelaeaceae) and the root of Wikstroemia indica (Thymelaeaceae), and 1 alkaloid isolated from Caulerpa racemosa (Caulerpaceae). The in vitro anticancer effects of these 28 natural compounds on 11 human cancer cell lines, including malignant melanoma, lung carcinoma, cervix epithelial adenocarcinoma, liver carcinoma, prostate adenocarcinoma and colorectal adenocarcinoma, were tested to set up an overall anticancer activity database for better understanding of the biological actions of Annonaceous plants. / Among the screened natural compounds, Annonaceous acetogenins (ACGs) exhibited outstanding anticancer efficacy. The cytotoxicities of ACGs to some cancer cell lines were even at nmol/l level. For instance, desacetyluvaricin (Dau), an ACG, was identified as a novel antiproliferative agent with a broad spectrum of inhibitions against the tested 11 human cancer cell lines with the IC₅₀ values ranging from 2.3 nM to 37.4 μM, and was especially cytotoxic to SW480 human colorectal carcinoma cells. Despite this potency, Dau was virtually nontoxic toward Hs68 human fibroblasts with an IC₅₀ value exceeding 247.5 μM. Further cell death mechanism studies showed that Dau could induce large amounts of superoxide production, which subsequently induced nuclear DNA fragmentation. DNA damage may inactivate the MEK/ERK signaling pathway and disturbed the expressions of cell cycle regulators such as Cyclin A and Cyclin E, which are S phase regulators, and E2F which is the G1/S checkpoint regulator. Thereafter, Dau arrested SW480 cells in S phase by promoting SW480 cells passing through the G₁/S boundary, and then accumulating in S phase. Finally, the SW480 cells underwent necrotic cell death. This mechanism study may provide a better understanding on the action mode of ACGs. / ACGs are derivatives of long chain fatty acids. Its structural diversity kindled our great interests in its structure-activity relationship (SAR). Therefore, we compared the cytotoxicities and cell cycle regulations of the 14 ACG compounds on two different human prostate cancer cell lines, LNCaP (p53 wild-type) and PC-3 (p53 null-type). Results showed that LNCaP cells were more sensitive to ACGs than PC-3 cells. This selectivity may be due to the presence of p53 tumor suppressor gene. Moreover, we found about SAR study that (1) the more THF rings existing in the core structure of ACGs, the more potent anticancer effects of ACGs would be; (2) for the adjacent bis-THF ACGs, stereo-structure with threo/trans/threo/trans/erythro configuration is generally more cytotoxic than the one with threo/trans/threo/trans/threo configuration; (3) both mono-THF ACGs and bis-THF ACGs inhibited LNCaP cells growth by G₁/G₀ phase arrest without any apoptosis induction; (4) mono-THF ACGs inhibited PC-3 cells growth by inducing apoptosis without cell cycle disturbance. However, the bis-THF ACGs could induce more apoptosis in PC-3 cells with partially cell cycle arrest. (5) the -OAc substituent group instead of -OH in the THF system would enhance the cytotoxicity efficacies of ACGs; (6) the double bond substituent would also enhance the anticancer effect. Our studies have proved several reported disciplines about the SAR of ACGs, and also proposed some new hypothesis. / Taken together, this study not only increased our understanding on the potent anticancer effects of ACG, but also provided valuable information on explaining its special cytotoxicities and the SAR properties through underling mechanism study. ACGs are a group of promising anticancer compounds with potent and steady activities. In the future work, we should further examine the in vivo anticancer effects and study more ACGs on their action modes to validate our hypothesis on their sensitivities to certain cancer cell lines. / 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. / Xue, Junyi. / Thesis (Ph.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 215-236). / Abstracts also in Chinese.

by Fok Ho Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 114-120). / Abstracts in English and Chinese. / Acknowledgements --- p.ii / Abstract --- p.iii / 槪論 --- p.v / List of Abbreviations --- p.vi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Toxicological studies on the effect of Ilex latifolia extract and its saponin-enriched Fraction --- p.19 / Chapter Chapter 3 --- Hypoglycemic effect of Ilex latifolia extract and its saponin-enriched fraction --- p.51 / Chapter Chapter 4 --- Hypolipidemic effect of Ilex latifolia extract and its saponin-enriched fraction --- p.78 / Chapter Chapter 5 --- Conclusion --- p.109 / References --- p.114

Liu, Yanan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 87-98). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese.

Ong Chik Ying Rose. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 181-195). / Abstracts in English and Chinese. / Acknoledgements --- p.i / List of Abbreviations --- p.ii / Abstract --- p.iii / Abstract in Chinese --- p.v / List of Publications and Abstracts --- p.vii / List of Figures --- p.viii / List of Tables --- p.xi / Contents --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Saponins --- p.3 / Chapter 1.1.1 --- Structure of saponins --- p.3 / Chapter 1.1.2 --- Occurrence of saponins --- p.7 / Chapter 1.1.3 --- Bioactivities of saponins --- p.8 / Chapter 1.2 --- Paris Polyphylla --- p.9 / Chapter 1.2.1 --- Chonglou --- p.11 / Chapter 1.3 --- Polyphyllin D --- p.12 / Chapter 1.4 --- Apoptosis --- p.15 / Chapter 1.4.1 --- Apoptosis and necrosis --- p.16 / Chapter 1.4.2 --- Initiation phase of apoptosis --- p.16 / Chapter 1.4.2.1 --- Extrinsic apoptotic pathway --- p.18 / Chapter 1.4.2.2 --- Intrinsic apoptotic pathway --- p.19 / Chapter 1.4.3 --- Execution phase of apoptosis --- p.19 / Chapter 1.4.4 --- Termination phase of apoptosis --- p.20 / Chapter 1.5 --- Multi-drug resistance (MDR) --- p.21 / Chapter 1.5.1 --- MDR mediated by decreased drug accumulation --- p.21 / Chapter 1.5.2 --- MDR mediated by enhanced anti-oxidant enzyme activities --- p.25 / Chapter 1.5.3 --- MDR mediated by enhanced detoxification of drugs --- p.25 / Chapter 1.5.4 --- MDR mediated by enhanced DNA repair system --- p.26 / Chapter 1.5.5 --- MDR mediated by altered apoptotic pathway --- p.26 / Chapter 1.5.6 --- Current strategies for overcoming multidrug resistance in cancer --- p.27 / Chapter 1.6 --- Hepatocellular carcinoma (HCC) --- p.30 / Chapter 1.7 --- Objectives of project --- p.32 / Chapter Chapter 2 --- Materials and Methods --- p.33 / Chapter 2.1 --- Materials --- p.34 / Chapter 2.1.1 --- Culture of Cells --- p.34 / Chapter 2.1.1.1 --- Cell lines --- p.34 / Chapter 2.1.1.2 --- Preservation of Cells --- p.35 / Chapter 2.1.2 --- Culture Media --- p.36 / Chapter 2.1.2.1 --- RPMI 1640 (Phenol Red Medium) --- p.36 / Chapter 2.1.2.2 --- RPMI 1640 (Phenol Red-free Medium) --- p.36 / Chapter 2.1.3 --- Buffers and Reagents --- p.37 / Chapter 2.1.3.1 --- Buffer for Common Use --- p.37 / Chapter 2.1.3.2 --- Reagents for Annexin-V-FITC/PI Assay --- p.37 / Chapter 2.1.3.3 --- Reagents for Western Blotting Analysis --- p.37 / Chapter 2.1.4 --- Chemicals --- p.40 / Chapter 2.1.4.1 --- Fluorescence Dyes --- p.40 / Chapter 2.1.4.2 --- Antibodies --- p.41 / Chapter 2.1.4.3 --- Other Chemicals --- p.42 / Chapter 2.1.5 --- Summary of chemicals used in this study --- p.43 / Chapter 2.2 --- Methods --- p.48 / Chapter 2.2.1 --- In vitro Cell Cytotoxicity Assay --- p.48 / Chapter 2.2.1.1 --- AlamarBlue Assay --- p.48 / Chapter 2.2.2 --- Flow Cytometry --- p.50 / Chapter 2.2.2.1 --- Analysis by Flow Cytometry --- p.50 / Chapter 2.2.2.2 --- Determination of Apoptotic and Late Apoptotic/Necrotic Cells with Annexin-V-FITC/PI Cytometric Analysis --- p.50 / Chapter 2.2.2.3 --- Determination of Mitochondrial Membrane Potential in cells --- p.51 / Chapter 2.2.2.4 --- Determination of Hydrogen Peroxide (H2O2) Release in cells --- p.52 / Chapter 2.2.2.5 --- Measurement of doxorubicin accumulation in cells --- p.53 / Chapter 2.2.2.6 --- Determination of P-glycoprotein expression level in cells --- p.54 / Chapter 2.2.2.7 --- Determination of mitochondrial depolarization and swellingin isolated mitochondria --- p.54 / Chapter 2.2.3 --- Methods involved in DNA sequencing of MDRl promoter --- p.56 / Chapter 2.2.3.1 --- DNA extraction --- p.56 / Chapter 2.2.3.2 --- DNA and Gel Band Purification --- p.56 / Chapter 2.2.3.3 --- Assessment of DNA amount --- p.57 / Chapter 2.2.3.4 --- Polymerase Chain Reaction --- p.57 / Chapter 2.2.3.5 --- Agarose Gel Electrophoresis --- p.59 / Chapter 2.2.3.6 --- Preparation for DNA sequencing --- p.59 / Chapter 2.2.4 --- Western Blotting Analysis --- p.61 / Chapter 2.2.4.1 --- Preparation of Proteins from Cells --- p.61 / Chapter 2.2.4.2 --- Preparation of proteins from isolated mitochondria --- p.63 / Chapter 2.2.4.3 --- Protein analysis with Western analysis --- p.63 / Chapter 2.2.5 --- Confocal laser scanning microscopy (Confocal microscopy) --- p.66 / Chapter 2.2.5.1 --- Analysis with confocal microscopy --- p.66 / Chapter 2.2.5.2 --- Determination of mitochondrial changes in cells by confocal microscopy --- p.66 / Chapter 2.2.5.3 --- Determination of lysosomal rupture in cells by confocal microscopy --- p.67 / Chapter 2.2.6 --- Mitochondrial isolation --- p.68 / Chapter Chapter 3 --- Results 一 Resistance Mechanisms in RHepG2 cells --- p.69 / Chapter 3.1 --- Resistance of RHepG2 cells towards various chemical agents --- p.70 / Chapter 3.1.1 --- RHepG2 cells are resistant to doxorubicin --- p.70 / Chapter 3.1.2 --- RHepG2 cells are resistant to taxol --- p.72 / Chapter 3.1.3 --- RHepG2 cells are resistant to valinomycin --- p.74 / Chapter 3.2 --- Resistance mechanism in RHepG2 cells --- p.76 / Chapter 3.2.1 --- Reduced doxorubicin accumulation is observed in RHepG2 cells --- p.76 / Chapter 3.2.2 --- More P-glycoproteins on the cell surface was observed in RHepG2 cells --- p.80 / Chapter 3.2.3 --- Inhibition of P-glycoprotein activity increased doxorubicin accumulation in RHepG2 cells --- p.82 / Chapter 3.2.4 --- HepG2 and RHepG2 cells contain the same P-glycoprotein promoter region --- p.86 / Chapter 3.2.5 --- RHepG2 over-expressed Bcl-2 --- p.91 / Chapter 3.2.6 --- HepG2 and RHepG2 cells had the same level of Bax protein --- p.93 / Chapter Chapter 4 --- Results - Effects of Polyphyllin D in HepG2 and RHepG2 cells --- p.95 / Chapter 4.1 --- Cytotoxicity of Polyphyllin D in HepG2 and RHepG2 cells --- p.96 / Chapter 4.1.1 --- Polyphyllin D exhibited cytotoxic effect in both HepG2 and RHepG2 cells --- p.96 / Chapter 4.2 --- Apoptotic mechanism caused by Polyphyllin D in HepG2 and RHepG2 cells --- p.93 / Chapter 4.2.1 --- Polyphyllin D caused apoptosis in HepG2 and RHepG2 cells --- p.98 / Chapter 4.2.2 --- Polyphyllin D did not activate caspase8 --- p.103 / Chapter 4.2.3 --- Polyphyllin D did not concentrate on the plasma membrane of cells --- p.106 / Chapter 4.2.4 --- Polyphyllin D did not change Bcl-2 level in HepG2 and RHepG2 cells --- p.109 / Chapter 4.2.5 --- Polyphyllin D treatment enhanced Bax protein expression in both HepG2 and RHepG2 cells --- p.111 / Chapter 4.2.6 --- Polyphyllin D caused cytochrome c and AIF release in HepG2 and RHepG2 cells --- p.113 / Chapter 4.2.7 --- Polyphyllin D induced mitochondrial membrane depolarization in HepG2 and RHepG2 cells --- p.118 / Chapter 4.2.8 --- Polyphyllin D caused mitochondrial swelling in HepG2 and clustering of mitochondriain RHepG2 cells --- p.122 / Chapter 4.2.9 --- "Polyphyllin D caused H202 release in HepG2 and RHepG2 cells, and the cytotoxic effects of Polyphyllin D could be reduced by NAC" --- p.127 / Chapter 4.2.10 --- Polyphyllin D caused permeabilization of lysosomes --- p.132 / Chapter 4.3 --- Site of action of Polyphyllin D in cells --- p.135 / Chapter 4.3.1 --- Purity of isolated mitochondria --- p.135 / Chapter 4.3.2 --- Polyphyllin D caused cytochrome c release from the HepG2 and RHepG2 isolated mitochondria --- p.137 / Chapter 4.3.3 --- Polyphyllin D induced mitochondrial depolarization in HepG2 and RHepG2 isolated mitochondria --- p.139 / Chapter 4.3.4 --- Polyphyllin D caused mitochondrial swelling in HepG2 and RHepG2 isolated mitochondria --- p.142 / Chapter 4.4 --- Resistance reversal effects of Polyphyllin D in RHepG2 cells --- p.144 / Chapter 4.4.1 --- Polyphyllin D increased doxorubicin accumulation in RHepG2 cells --- p.144 / Chapter 4.4.2 --- P-glycoprotein expression was not down-regulated after Polyphyllin D treatment --- p.146 / Chapter 4.4.3 --- Co-treatment of doxorubicin with Polyphyllin D had enhanced cytotoxic effect --- p.148 / Chapter Chapter 5 --- Discussion - Resistance mechanisms in RHepG2 cells --- p.150 / Chapter 5.1 --- Resistance of RHepG2 cells towards various chemical reagents --- p.151 / Chapter 5.2 --- Resistance mechanisms in RHepG2 cells --- p.154 / Chapter Chapter 6 --- Discussion - Effects of Polyphyllin D in HepG2and RHepG2 cells --- p.159 / Chapter 6.1 --- Cytotoxicity of Polyphyllin D in HepG2 and RHepG2 cells --- p.160 / Chapter 6.2 --- Apoptotic mechanisms caused by Polyphyllin D in HepG2 and RHepG2 cells --- p.162 / Chapter 6.3 --- Site of action of Polyphyllin D in HepG2 andRHepG2 cells --- p.172 / Chapter 6.4 --- Resistance reversal effects of Polyphyllin D in RHepG2 cells --- p.175 / Chapter Chapter 7 --- Future Perspectives --- p.177 / Chapter Chapter 8 --- Conclusion --- p.179 / References

Chan Wai Sing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 199-210). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.vi / CHINESE ABSTRACT --- p.x / TABLE OF CONTENTS --- p.xiii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- What is hematopoiesis? --- p.1 / Chapter 1.1.1 --- The development of hematopoietic progenitor cells --- p.1 / Chapter 1.1.2 --- The regulation of hematopoiesis by environmental factors --- p.4 / Chapter 1.1.3 --- The regulation of hematopoiesis by transcription factors --- p.7 / Chapter 1.2 --- Leukemia --- p.9 / Chapter 1.2.1 --- Classification of leukemia --- p.11 / Chapter 1.2.2 --- Pathology and etiology of leukemia --- p.15 / Chapter 1.2.2.1 --- Inheritance --- p.15 / Chapter 1.2.2.2 --- Environmental factors --- p.16 / Chapter 1.2.2.3 --- Virus infection --- p.17 / Chapter 1.2.3 --- Genetics of leukemia --- p.17 / Chapter 1.2.3.1 --- Point mutations --- p.17 / Chapter 1.2.3.2 --- Translocations --- p.18 / Chapter 1.2.3.3 --- Gene and chromosomal deletions --- p.18 / Chapter 1.2.3.4 --- Chromosomal duplication or gene amplification --- p.18 / Chapter 1.2.4 --- Current therapeutic strategies for leukemia --- p.20 / Chapter 1.2.4.1 --- Chemotherapy --- p.20 / Chapter 1.2.4.2 --- Stem cell transplantation --- p.21 / Chapter 1.2.4.3 --- Immunotherapy --- p.22 / Chapter 1.2.4.4 --- Gene therapy --- p.23 / Chapter 1.2.5 --- Novel approaches for the treatment of leukemia --- p.23 / Chapter 1.2.5.1 --- Differentiation therapy of leukemia --- p.24 / Chapter 1.2.5.2 --- Induction of apoptosis in the treatment of leukemia --- p.25 / Chapter 1.3 --- Topoisomerase-targeting agents --- p.27 / Chapter 1.3.1 --- What is topoisomerase? --- p.27 / Chapter 1.3.2 --- Structures and action mechanisms of Top I- and Top II-targeting agents --- p.28 / Chapter 1.3.3 --- Anti-tumor activities of topoisomerase-targeting agents --- p.37 / Chapter 1.4 --- Aims and scopes of this investigation --- p.39 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.42 / Chapter 2.1.1 --- Mice --- p.42 / Chapter 2.1.2 --- Cell lines --- p.42 / Chapter 2.1.3 --- "Cell culture medium, buffers and other reagents" --- p.43 / Chapter 2.1.4 --- Radioisotope and scintillation fluid --- p.47 / Chapter 2.1.5 --- Reagents and buffers for flow cytometry --- p.48 / Chapter 2.1.6 --- Antibodies for flow cytometry --- p.50 / Chapter 2.1.7 --- Recombinant cytokines --- p.51 / Chapter 2.1.8 --- Reagents for DNA extraction --- p.52 / Chapter 2.1.9 --- Reagents for total RNA isolation --- p.53 / Chapter 2.1.10 --- Reagents and buffers for RT-PCR --- p.54 / Chapter 2.1.11 --- Reagents and buffers for gel electrophoresis --- p.58 / Chapter 2.1.12 --- Reagents and buffers for Western blot analysis --- p.59 / Chapter 2.1.13 --- Reagents for measuring caspase activity --- p.64 / Chapter 2.2 --- Methods --- p.67 / Chapter 2.2.1 --- Culture of the leukemia cell lines --- p.67 / Chapter 2.2.2 --- Isolation and preparation of normal hematopoietic cells --- p.67 / Chapter 2.2.3 --- [3 H]-TdR proliferation assay --- p.68 / Chapter 2.2.4 --- Determination of cell viability --- p.69 / Chapter 2.2.5 --- Assay for anti-leukemic activity in vivo --- p.70 / Chapter 2.2.6 --- Assessment of differentiation-associated characteristics --- p.71 / Chapter 2.2.7 --- Assays for apoptosis --- p.73 / Chapter 2.2.8 --- Cell cycle analysis (DNA content evaluation) --- p.75 / Chapter 2.2.9 --- Gene expression study --- p.75 / Chapter 2.2.10 --- Protein expression study --- p.79 / Chapter 2.2.11 --- Measurement of caspase activity --- p.82 / Chapter 2.2.12 --- Statistical analysis 一 --- p.83 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-PROLIFERATIVE EFFECT OF TOPOISOMERASE-TARGETING AGENTS ON LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.84 / Chapter 3.2 --- Results --- p.86 / Chapter 3.2.1 --- The anti-proliferative effect of topoisomerase-targeting agents on human and murine leukemia cells in vitro --- p.86 / Chapter 3.2.2 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the clonogenicity of the murine myeloid leukemia WEHI-3B JCS cells in vitro --- p.105 / Chapter 3.2.3 --- Effects of 10-hydroxy-camptothecin and etoposide phosphate on the tumorigenicity and proliferation of the murine myeloid leukemia WEHI-3B JCS cells invivo --- p.106 / Chapter 3.2.4 --- Cytotoxic effect of 10-hydroxy-camptothecin and etoposide phosphate on normal hematopoietic cells and WEHI-3B JCS cells in vitro --- p.109 / Chapter 3.2.5 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the cell cycle kinetics of WEHI-3B JCS cells --- p.114 / Chapter 3.2.6 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of cell cycle-regulatory genes in the murine myeloid leukemia WEHI-3B JCS cells --- p.116 / Chapter 3.2.7 --- Combination effect of 10-hydroxy-camptothecin or etoposide phosphate with cytokines on the proliferation of the murine myeloid leukemia WEHI-3B JCS cells --- p.123 / Chapter 3.3 --- Discussion --- p.127 / Chapter CHAPTER 4: --- STUDIES ON THE DIFFERENTIATION-INDUCING EFFECT OF 10-HYDROXY-CAMPTOTHECIN AND ETOPOSIDE PHOSPHATE ON THE MURINE MYELOID LEUKEMIA WEHI-3B JCS CELLS / Chapter 4.1 --- Introduction --- p.132 / Chapter 4.2 --- Results --- p.134 / Chapter 4.2.1 --- Morphological changes in the murine myeloid leukemia WEHI-3B JCS cells treated with 10-hydroxy-camptothecin and etoposide phosphate --- p.134 / Chapter 4.2.2 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the size and granularity of the murine myeloid leukemia WEHI-3B JCS cells --- p.138 / Chapter 4.2.3 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the plastic adhering property of the murine myeloid leukemia WEHI-3B JCS cells --- p.140 / Chapter 4.2.4 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the NBT-reducing activity of the murine myeloid leukemia WEHI-3B JCS cells --- p.142 / Chapter 4.2.5 --- Surface antigen immunophenotyping of the murine myeloid leukemia WEHI-3B JCS cells treated with 10-hydroxy- camptothecin and etoposide phosphate --- p.145 / Chapter 4.2.6 --- Induction of non-specific esterase activity in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.152 / Chapter 4.3 --- Discussion --- p.154 / Chapter CHAPTER 5: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF 10-HYDROXY-CAMPTOTHECIN AND ETOPOSIDE PHOSPHATE ON THE MURINE MYELOID LEUKEMIA WEHI-3B JCS CELLS / Chapter 5.1 --- Introduction --- p.157 / Chapter 5.2 --- Results --- p.160 / Chapter 5.2.1 --- Induction of nuclear disintegration in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy-camptothecin and etoposide phosphate --- p.160 / Chapter 5.2.2 --- Induction of DNA fragmentation in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy-camptothecin and etoposide phosphate --- p.162 / Chapter 5.2.3 --- Induction of phosphatidylserine translocation in murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.167 / Chapter 5.2.4 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of apoptosis-regulatory genes in the murine myeloid leukemia WEHI-3B JCS cells --- p.171 / Chapter 5.2.5 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of apoptosis-regulatory proteins in the murine myeloid leukemia WEHI-3B JCS cells --- p.177 / Chapter 5.2.6 --- Induction of mitochondrial membrane depolarization in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.179 / Chapter 5.2.7 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on caspase activity in the murine myeloid leukemia WEHI-3B JCS cells --- p.181 / Chapter 5.2.8 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate intracellular Ca2+ level in the murine myeloid leukemia WEHI-3B JCS cells --- p.186 / Chapter 5.3 --- Discussion --- p.189 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.192 / REFERENCES --- p.199

Chu Ka-wing, Kevin. / "June 2001." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (p. 194-215) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Christopher John Haines. / Thesis (M.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 239-279). / LIST OF TABLES --- p.xviii / LIST OF FIGURES --- p.xxi / LIST OF ABBREVIATIONS --- p.xxii / Chapter CHAPTER1 --- INTRODUCTION --- p.1 / Problems related to the menopause / Research plan / Chapter CHAPTER2 --- OVERVIEW --- p.15 / Introduction / Atherosclerosis and the lipid profile / Coronary artery disease and lipid abnormalities in women / "Exogenous oestrogens, progestogens and coronary artery disease " / Lipoprotein (a) / Chapter CHAPTER3 --- GENERAL METHODOLOGY --- p.134 / Recruitment of cases / Pharmacokinetics / Data collection and analysis of samples / Ethical considerations / Chapter CHAPTER4 --- STUDY I -THE SHORT TERM EFFECTS OF ORAL OESTROGEN --- p.157 / Crossover analysis of effects of oral oestrogen on lipoprotein (a) and other lipoproteins / Relationship between lipoprotein (a) and other lipids and lipoproteins / Chapter CHAPTER5 --- STUDY II -THE SUSTAINED EFFECTS OF ORAL OESTROGEN --- p.186 / Analysis of prolonged effects of oral oestrogen on lipoprotein (a) and other lipids and lipoproteins / Chapter CHAPTER6 --- STUDY III -THE EFFECTS OF COMBINED CYCLICAL HORMONE REPLACEMENT THERAPY --- p.196 / Analysis of effect of combined cyclical hormone replacement therapy on lipoprotein (a) and other lipids and lipoproteins / Comparison between sampling during oestrogen alone and combined phase of treatment / Chapter CHAPTER7 --- STUDY IV -THE EFFECTS OF PERCUTANEOUS OESTROGEN --- p.214 / Analysis of effect of percutaneous on lipoprotein (a) and other lipids and lipoproteins / Chapter CHAPTER8 --- SUMMARY AND CONCLUSIONS --- p.228 / BIBLIOGRAPHY --- p.239

Liu, Xiaozhuo. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 156-169). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese (摘要） --- p.iv / Acknowledgments --- p.vi / List of Abbreviations --- p.vii / Table of Contents --- p.xiv / Chapter Chapter One: --- General Introduction / Chapter 1.1 --- Hematopoiesis and Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis --- p.1 / Chapter 1.1.2 --- Leukemia --- p.6 / Chapter 1.1.2.1 --- An Overview of Leukemia --- p.6 / Chapter 1.1.2.2 --- Classification and Epidemiology of Leukemia --- p.8 / Chapter 1.1.2.3 --- Conventional Approaches to Leukemia Therapy --- p.12 / Chapter 1.1.2.4 --- Novel Approaches to Leukemia Therapy --- p.15 / Chapter 1.2 --- Sophoraflavanone G: A Bioactive Compound Isolated from Kushen --- p.18 / Chapter 1.2.1 --- An Overview of Kushen: A Traditional Chinese Medicine --- p.19 / Chapter 1.2.2 --- An Overview of Lavandulyl Flavanones --- p.22 / Chapter 1.2.3 --- Historical Development and Occurrence of Sophoraflavanone G --- p.24 / Chapter 1.2.4 --- Biological Activities of Sophoraflavanone G --- p.25 / Chapter 1.2.4.1 --- Anti-microbial and Insecticidal Activities --- p.25 / Chapter 1.2.4.2 --- Anti-tumor Activities --- p.26 / Chapter 1.2.4.3 --- Pharmacodynamics of Sophoraflavanone G --- p.27 / Chapter 1.3 --- Objectives and Scopes of the Present Study --- p.30 / Chapter Chapter Two: --- Materials and Methods / Chapter 2.1 --- Materials --- p.32 / Chapter 2.1.1 --- Animals --- p.32 / Chapter 2.1.2 --- Cell lines --- p.32 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.34 / Chapter 2.1.4 --- Reagents and Buffers for Flow Cytometry --- p.37 / Chapter 2.1.5 --- Reagents for DNA Extraction --- p.39 / Chapter 2.1.6 --- Reagents for Measuring Caspase Activity --- p.40 / Chapter 2.1.7 --- "Reagents, Buffers and Materials for Western Blotting" --- p.43 / Chapter 2.2 --- Methods --- p.48 / Chapter 2.2.1 --- Extraction and Isolation of Sophoraflavanone G from Kushen --- p.48 / Chapter 2.2.2 --- Culture of Tumor Cell Lines --- p.49 / Chapter 2.2.3 --- "Isolation, Preparation and Culturing of Human Peripheral Blood Leukocytes and Murine Bone Marrow Cells" --- p.50 / Chapter 2.2.4 --- Assays for Anti-proliferation and Cytotoxicity --- p.51 / Chapter 2.2.5 --- Determination of Anti-leukemic Activity In Vivo (In Vivo Tumorigenicity Assay) --- p.52 / Chapter 2.2.6 --- Cell Cycle Analysis by Flow Cytometry --- p.53 / Chapter 2.2.7 --- Measurement of Apoptosis-induced Activities --- p.54 / Chapter 2.2.8 --- Protein Expression Study --- p.59 / Chapter 2.2.9 --- Assessment of Differentiation-associated Characteristics --- p.64 / Chapter 2.2.10 --- Statistical Analysis --- p.65 / Chapter Chapter Three: --- Studies on the Anti-proliferative Effect of Sophoraflavanone G on Human Myeloid Leukemia Cells / Chapter 3.1 --- Introduction --- p.66 / Chapter 3.2 --- Results --- p.69 / Chapter 3.2.1 --- Structure Identification of Sophoraflavanone G Isolated from Sophora flavescens --- p.69 / Chapter 3.2.2 --- Anti-proliferative Activity of Sophoraflavanone G on Various Myeloid Leukemia Cell Lines --- p.72 / Chapter 3.2.3 --- Effect of Sophoraflavanone G on the Viability of the Human Promyelocytic Leukemia HL-60 Cells --- p.80 / Chapter 3.2.4 --- Cytotoxic Effect of Sophoraflavanone G on Primary Normal Cells In Vitro --- p.83 / Chapter 3.2.5 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Sophoraflavanone G on the Human Promyelocytic Leukemia HL-60 Cells --- p.85 / Chapter 3.2.6 --- Effect of Sophoraflavanone G on the In Vivo Tumorigenicity of the HL-60 Cells --- p.88 / Chapter 3.2.7 --- Effect of Sophoraflavanone G on the Cell Cycle Profile of the HL-60 cells In Vitro --- p.90 / Chapter 3.2.8 --- Effect of Sophoraflavanone G on the Expression of Cell Cycle-regulatory Proteins in the HL-60 Cells --- p.93 / Chapter 3.2.9 --- Anti-proliferative Effect of Sophoraflavanone G on Multidrug-resistant (MDR) Leukemia Cell Line HL-60/MX2 Cells --- p.95 / Chapter 3.3 --- Discussion --- p.101 / Chapter Chapter Four: --- Studies on the Apoptosis- and Differentiation-inducing Activities of Sophoraflavanone G on Human Myeloid Leukemia Cells / Chapter 4.1 --- Introduction --- p.109 / Chapter 4.2 --- Results --- p.114 / Chapter 4.2.1 --- Induction of DNA Fragmentation in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G --- p.114 / Chapter 4.2.2 --- Induction of Phosphatidylserine Externalization in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G as Detected by Annexin V-GFP and PI Double Staining Method --- p.116 / Chapter 4.2.3 --- Effects of Sophoraflavanone G on the Caspase Activities in the Human Promyelocytic Leukemia HL-60 Cells --- p.119 / Chapter 4.2.4 --- Induction of Mitochondrial Membrane Depolarization in the Human Promyelocytic Leukemia HL-60 Cells by Sophoraflavanone G --- p.124 / Chapter 4.2.5 --- Involvement of Bcl-2 Family Members in Sophoraflavanone G-induced Apoptosis in the Human Promyelocytic Leukemia HL-60 Cells --- p.128 / Chapter 4.2.6 --- Effects of Sophoraflavanone G on the Induction of Reactive Oxygen Species in the Human Promyelocytic Leukemia HL-60 Cells --- p.131 / Chapter 4.2.7 --- Effect of Sophoraflavanone G on the Intracellular Ca2+ Level in the Human Promyelocytic Leukemia HL-60 Cells --- p.134 / Chapter 4.2.8 --- Morphological Studies on the Sophoraflavanone G-treated Human Promyelocytic Leukemia HL-60 Cells --- p.136 / Chapter 4.2.9 --- Effect of Sophoraflavanone G on the NBT Reducing Activity of the Human Promyelocytic Leukemia HL-60 Cells --- p.138 / Chapter 4.3 --- Discussion --- p.140 / Chapter Chapter Five: --- Conclusions and Future Perspectives --- p.148 / References --- p.156