Lo, Yu Ching. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 79-82). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abbreviations --- p.ii / Abstract --- p.iii / Table of Contents --- p.v / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Basic Theory of FTIR --- p.1 / Chapter 1.1.1 --- Infrared Spectroscopy --- p.1 / Chapter 1.1.2 --- Dispersive Infrared Spectrometer --- p.2 / Chapter 1.1.3 --- Fourier Transform Infrared Spectrometer (FTIR) --- p.3 / Chapter 1.1.4 --- Advantages of FTIR --- p.5 / Chapter 1.1.4.1 --- Multiplex advantage --- p.5 / Chapter 1.1.4.2 --- Throughput advantage --- p.5 / Chapter 1.2 --- Traditional Chinese Medicine (TCM) --- p.6 / Chapter 1.3 --- Identification of Traditional Chinese Herbal Medicine --- p.8 / Chapter 1.3.1 --- Traditional method for the identification of TCM --- p.8 / Chapter 1.3.2 --- Instrumental method for the identification of TCM --- p.9 / Chapter 1.3.2.1 --- Identification of TCM using fingerprint method --- p.9 / Chapter 1.3.3 --- Identification of TCM using FTIR fingerprint method --- p.10 / Chapter 1.4 --- Objective --- p.11 / Chapter Chapter 2 --- Experimental / Chapter 2.1 --- Outline of the Method --- p.12 / Chapter 2.2 --- Reagents and Glassware --- p.13 / Chapter 2.3 --- Instrumentation --- p.13 / Chapter 2.4 --- Library Search Program --- p.13 / Chapter 2.5 --- Samples --- p.14 / Chapter 2.6 --- Sample Pretreatment --- p.16 / Chapter 2.7 --- Extraction of Ingredients --- p.16 / Chapter 2.8 --- Preparation of KBr Pellet --- p.17 / Chapter 2.9 --- IR Spectrum Measurement --- p.17 / Chapter 2.10 --- Data Processing --- p.18 / Chapter 2.11 --- IR Databases --- p.18 / Chapter 2.12 --- Reproducibility of Extraction --- p.29 / Chapter Chapter 3 --- Application of Library Search Techniques - Results and Disscussion / Chapter 3.1 --- Introduction --- p.31 / Chapter 3.2 --- Euclidean Search Method --- p.32 / Chapter 3.2.1 --- Similarly score of reference spectra --- p.32 / Chapter 3.2.2 --- Similarity score of known sample spectra --- p.37 / Chapter 3.3 --- Soft Independent Modeling of Class Analogy (SIMCA) --- p.45 / Chapter 3.3.1 --- Verification Diagnostic Report of the Reference flowers --- p.47 / Chapter 3.3.2 --- Classification of Flowers --- p.51 / Chapter 3.4 --- Performance Limitation --- p.75 / Chapter 3.4.1 --- Euclidean Search Method --- p.75 / Chapter 3.4.2 --- SIMCA --- p.75 / Chapter 3.5 --- Conclusion --- p.77 / References --- p.79

Tang Sze-Wan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 104-118). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese) --- p.iii / Acknownledgement --- p.iv / List of Abbreviations --- p.v / List of Tables --- p.vii / List of Figures --- p.viii / Table of Contents --- p.xi / Chapter Chapter 1 - --- Introduction / Chapter 1.1 --- Trichosanthes spp --- p.1 / Chapter 1.1.1 --- Use of Trichosanthes --- p.2 / Chapter 1.1.2 --- Trichosanthin --- p.2 / Chapter 1.1.3 --- Karasurin --- p.5 / Chapter 1.1.4 --- Ribosome Inactivating Proteins --- p.6 / Chapter 1.1.5 --- Immunosuppresion --- p.7 / Chapter 1.1.6 --- Anti-Cancer Activity --- p.8 / Chapter 1.1.7 --- Miscellaneous Uses --- p.8 / Chapter 1.2 --- Cancer --- p.9 / Chapter 1.2.1 --- Oncogenes --- p.10 / Chapter 1.2.2 --- Tumor-Suppressor Genes --- p.11 / Chapter 1.2.3 --- Stability Genes --- p.12 / Chapter 1.2.4 --- Types of Cancer --- p.13 / Chapter 1.2.5 --- Cancer Therapy --- p.13 / Chapter 1.2.6 --- Apoptosis --- p.14 / Chapter 1.3 --- Chronic Myelogenous Leukemia (CML) --- p.17 / Chapter 1.3.1 --- Philadelphia Chromosome and BCR-ABL gene --- p.18 / Chapter 1.3.2 --- Treatment of CML --- p.20 / Chapter 1.4 --- Dendritic Differentiation of LC976 on K-562 --- p.20 / Chapter 1.4.1 --- Dendritic Cells --- p.21 / Chapter 1.4.2 --- Cancer Vaccine Development of Leukemia --- p.22 / Chapter 1.4.3 --- Dendritic differentiation of K-562 cells --- p.23 / Chapter 1.5 --- Perspective of the Project --- p.23 / Chapter Chapter 2 - --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Chemicals and Reagents --- p.25 / Chapter 2.1.2 --- Bioassay Kits --- p.26 / Chapter 2.1.3 --- Human Cell Lines --- p.26 / Chapter 2.1.4 --- Lab Wares and Equipments --- p.28 / Chapter 2.2 --- Extraction of LC9 --- p.76 / Chapter 2.2.1 --- Chemical Properties of the Lead Compound --- p.28 / Chapter 2.2.2 --- Crude Extraction of Trichosanthes sp --- p.29 / Chapter 2.2.3 --- Purification by Reversed-Phase Column --- p.29 / Chapter 2.2.4 --- Lyophilization and Preparation of LC976 --- p.31 / Chapter 2.3 --- Anti-Proliferation Effect of LC976 on Human Cell Lines / Chapter 2.3.1 --- Maintenance of Cell Lines --- p.32 / Chapter 2.3.2 --- MTT Assay --- p.32 / Chapter 2.3.3 --- BrdU Cell Proliferation ELISA --- p.34 / Chapter 2.4 --- Apoptosis Induction on K-5 --- p.62 / Chapter 2.4.1 --- PI Staining --- p.35 / Chapter 2.4.2 --- Annexin V-FITC FACS Analysis --- p.36 / Chapter 2.4.3 --- Caspase Activation --- p.37 / Chapter 2.5 --- Effect on Normal Human Lymphocytes / Chapter 2.5.1 --- Preparation of Human Normal Lymphocytes --- p.38 / Chapter 2.5.2 --- MTT Cell Viability Assay --- p.38 / Chapter 2.5.3 --- PI Staining --- p.39 / Chapter 2.5.4 --- Annexin V-FITC FACS Analysis --- p.39 / Chapter Chapter 3 - --- Results / Chapter 3.1 --- Extraction of LC976 --- p.40 / Chapter 3.2 --- LC976 Inhibited Proliferation of Human Cell Lines / Chapter 3.2.1 --- MTT Assay --- p.41 / Chapter 3.2.2 --- BrdU Cell Proliferation ELISA --- p.52 / Chapter 3.3 --- LC976 Induced Apoptosis in K-562 Cells / Chapter 3.3.1 --- PI Staining --- p.63 / Chapter 3.3.2 --- Annexin V-FITC FACS Analysis --- p.70 / Chapter 3.3.3 --- Caspase Activation --- p.73 / Chapter 3.4 --- Effect on Normal Human Lymphocytes / Chapter 3.4.1 --- MTT Cell Viability Assay --- p.76 / Chapter 3.4.2 --- PI Staining --- p.78 / Chapter 3.4.3 --- Annexin V-FITC FACS Analysis --- p.82 / Chapter Chapter 4 - --- Discussion / Chapter 4.1 --- Extraction of LC976 --- p.85 / Chapter 4.2 --- LC976 Inhibited Proliferation of Human Cell Lines / Chapter 4.2.1 --- MTT Assay --- p.86 / Chapter 4.2.2 --- BrdU Cell Proliferation ELISA --- p.88 / Chapter 4.3 --- LC976 induced Apoptosis in K-562 Cells / Chapter 4.3.1 --- PI Staining --- p.90 / Chapter 4.3.2 --- Annexin V-FITC Analysis --- p.95 / Chapter 4.3.3 --- Caspase Activation --- p.96 / Chapter 4.4 --- Effect on Normal Human Lymphocytes / Chapter 4.4.1 --- MTT Cell Viability Assay --- p.98 / Chapter 4.4.2 --- PI Staining --- p.99 / Chapter 4.4.3 --- Annexin V-FITC FACS Analysis --- p.100 / Chapter 4.5 --- Conclusion --- p.103 / Reference --- p.104

免疫系统包括一系列的免疫细胞和激活免疫反应的调节因子，正常的免疫系统可以保护机体免受各种病原体的侵害。免疫系统失调会导致各种各样的免疫性疾病，例如过敏，自身免疫性疾病和肿瘤。子宮内膜异位症是妇科常见慢性疾病之一，其特点为子宮内膜腺体和间质种植在子宮体以外部位。倒流的子宮内膜组织成功逃逸免疫系统的监控是成功种植的必要条件，然而其病理机制尚未研究清楚。 / 本研究探索了子宮内膜异位症小鼠模型24小时内腹腔免疫细胞的变化。我们发现大部分免疫细胞在子宮内膜组织移植24小时内显著降低，然而骨髓源性抑制细胞（MDSC）的比例大幅度增高。子宮内膜移植后6小时内已经有大量MDSC到达腹腔，并在子宮内膜粘附增生阶段都维持在高水平。子宮内膜异位诱导的腹腔MDSC包括两个细胞亚型，CD11b⁺Ly-6G⁺Ly-6C{U+02E1}{U+1D52}{U+02B7}粒细胞型MDSC (G-MDSC)　和CD11b⁺Ly-6G-Ly-6C{U+02B0}{U+2071}{U+1D4D}{U+02B0}单核细胞型MDSC (-MDSC)，二者都能够显著抑制T细胞增生，表明MDSC在早期子宮内膜异位症发展过程中介导了免疫抑制的发生。用Gr-1抗体去除子宮内膜异位症小鼠模型中的MDSC后，异位病灶的生长相比对照组受到显著抑制。同时，粒细胞集落刺激因子（G-CSF）在内膜组织移植后6小时内也明显增高，体内补充G-CSF会引起正常小鼠骨髓和周围血中MDSC的比例增高，这表明G-CSF可以诱导MDSC在骨髓中增生，并且在周围血中聚集。然而，G-CSF不能引起腹腔中MDSC增多，提示还有其他的因子参与了MDSC的趋化反应。 / 本研究表明MDSC在子宮内膜异位症发生发展过程中很可能通过类似于抑制肿瘤免疫反应的免疫抑制作用，而推动子宮内膜免疫逃逸过程，最终导致异位子宮内膜存活并生长。因此，MDSC可能成为研发子宮内膜异位症新治疗方案的靶点。 / Normal immune system keeps human bodies remain intact and unharmed in the midst of a vast universe of pathogens. The immune system includes a series of cells and molecules which worked together to initiate immune response. The dysfunction of immune cells or modulators results in various diseases, such as allergic diseases, autoimmune diseases and tumors. Endometriosis is a chronic gyneacological disorder characterized by the implantation of endometrial glands and stroma outside the uterus. The successful immune escape of retrograded endometrial cell from immune surveillance is one of the key steps in the development of endometriosis. The underlying mechanism is still unknown. / In this study, we observed the profiles of peritoneal immune cells and cytokines within 24 hours (h) after the transplantation of endometrial fragments in peritoneal cavity in mice. Most of the peritoneal immune cells decreased after the transplantation, except myeloid derived suppressor cells (MDSC) were significantly increased within 24 h. MDSC was increased in the peritoneal cavity as early as 6 h and maintained at high level when the endometrial fragments attached and proliferated in the following days. The increased MDSC have CD11b⁺Ly-6G⁺Ly-6C{U+02E1}{U+1D52}{U+02B7} granulocytic MDSC (G-MDSC) and CD11b⁺Ly-6G-Ly-6C{U+02B0}{U+2071}{U+1D4D}{U+02B0} monocytic MDSC (M-MDSC) phenotypes. Isolated peritoneal MDSC significantly suppressed T cell proliferation and activated arginase activity, suggesting the immunosuppression of MDSC during early development of endometriosis. Depletion of MDSC by Gr-1 antibody treatment significantly reduced the growth and development of endometriosis. Concurrently, granulocyte colony stimulating factor (G-CSF) was significantly increased as early as 6 h after transplantation. Supplementation of G-CSF in control mice increased MDSC percentage in bone marrow and peripheral blood, but not in peritoneal cavity. This suggests that G-CSF is responsible to induce the proliferation of MDSC in bone marrow and accumulation of MDSC in peripheral blood, but not recruitment into peritoneal cavity. Additional cytokines or chemokines may involve the mobilization of MDSC from peripheral blood to peritoneal cavity. / Our study suggested that MDSC may play an essential role in the immune escape and promote endometrial survival during early development of endometriosis, probably through the similar immunosuppression mechanisms as in cancer. MDSC may be a novel target to develop highly effective therapeutics for treatment of endometriosis. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhang, Tao. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 102-128). / Abstracts also in Chinese. / Acknowledgements --- p.i / Publications --- p.iii / Academic Awards --- p.iii / List of tables and figures --- p.ix / List of abbreviations --- p.ix / Abstract --- p.xv / Chapter Chapter 1 --- Endometriosis --- p.1 / Chapter 1.1 --- Epidemiology --- p.1 / Chapter 1.1.1 --- Prevalence --- p.1 / Chapter 1.1.2 --- Risk factors --- p.2 / Chapter 1.1.2.1 --- Menstrual and reproductive factors --- p.2 / Chapter 1.1.2.2 --- Genetics --- p.3 / Chapter 1.1.2.3 --- Physical and habitual factors --- p.6 / Chapter 1.1.2.4 --- Environmental toxins --- p.6 / Chapter 1.1.2.5 --- Immune comorbidity --- p.7 / Chapter 1.2 --- Pathogenesis --- p.8 / Chapter 1.2.1 --- Retrograde menstruation --- p.8 / Chapter 1.2.2 --- Lymphatic or vascular spread --- p.9 / Chapter 1.2.3 --- Coelomic metaplasia and induction theory --- p.10 / Chapter 1.3 --- Pathophysiology --- p.11 / Chapter 1.3.1 --- Dysfunction of immune system --- p.11 / Chapter 1.3.2 --- Attachment and invasion --- p.12 / Chapter 1.3.3 --- Angiogenesis --- p.12 / Chapter 1.3.4 --- Oxidative stress --- p.13 / Chapter 1.4 --- Symptoms --- p.13 / Chapter 1.4.1 --- Pelvic pain --- p.14 / Chapter 1.4.2 --- Dysmenorrhea --- p.15 / Chapter 1.4.3 --- Dyspareunia --- p.15 / Chapter 1.4.4 --- Infertility --- p.16 / Chapter 1.5 --- Diagnosis --- p.16 / Chapter 1.5.1 --- Laparoscopy --- p.17 / Chapter 1.5.2 --- Laboratory tests --- p.17 / Chapter 1.5.3 --- Imaging --- p.18 / Chapter 1.5.4 --- Therapeutic diagnosis --- p.19 / Chapter 1.6 --- Treatment --- p.19 / Chapter 1.6.1 --- Hormonal therapy --- p.19 / Chapter 1.6.1.1 --- Combined oral contraceptives --- p.20 / Chapter 1.6.1.2 --- Progestogens --- p.20 / Chapter 1.6.1.3 --- Danazol --- p.21 / Chapter 1.6.1.4 --- GnRH Agonists --- p.22 / Chapter 1.6.1.5 --- Gestrinone --- p.23 / Chapter 1.6.2 --- Surgical treatment --- p.23 / Chapter 1.6.2.1 --- Conservative surgery --- p.24 / Chapter 1.6.2.2 --- Definitive surgery --- p.24 / Chapter 1.6.3 --- Treatment of endometriosis-related infertility --- p.25 / Chapter 1.6.4 --- New potential therapies --- p.25 / Chapter 1.6.4.1 --- Angiogenesis inhibitors --- p.26 / Chapter 1.6.4.2 --- Antioxidant therapy --- p.26 / Chapter 1.6.4.3 --- Immunomodulatory therapy --- p.27 / Chapter 1.6.4.3.1 --- Pentoxifylline --- p.27 / Chapter 1.6.4.3.2 --- TNF-α inhibitor --- p.28 / Chapter 1.6.4.3.3 --- Loxoribine --- p.28 / Chapter 1.6.4.4 --- Others --- p.29 / Chapter 1.7 --- Summary --- p.29 / Chapter Chapter 2 --- Immunology of Endometriosis --- p.30 / Chapter 2.1 --- Cell-mediated immunity --- p.30 / Chapter 2.1.1 --- Monocytes/Macrophages --- p.31 / Chapter 2.1.2 --- Dendritic cells --- p.32 / Chapter 2.1.3 --- T lymphocyte --- p.34 / Chapter 2.1.4 --- Natural killer cells --- p.34 / Chapter 2.1.5 --- T regulatory cells --- p.35 / Chapter 2.2 --- Humoral immunity --- p.37 / Chapter 2.3 --- Cytokines and growth factors --- p.39 / Chapter 2.3.1 --- Interleukin-1 --- p.39 / Chapter 2.3.2 --- Interleukin-6 --- p.40 / Chapter 2.3.3 --- Interleukin-8 --- p.41 / Chapter 2.3.4 --- Interleukin-12 --- p.42 / Chapter 2.3.5 --- Monocyte chemotactic protein-1 --- p.43 / Chapter 2.3.6 --- RANTES --- p.43 / Chapter 2.3.7 --- Tumor necrosis factor-α --- p.44 / Chapter 2.3.8 --- Vascular endothelial growth factor --- p.45 / Chapter 2.3.9 --- Growth factors --- p.46 / Chapter 2.3.9.1 --- Insulin-like growth factor --- p.46 / Chapter 2.3.9.2 --- Transforming growth factor beta --- p.46 / Chapter 2.3.9.3 --- Platelet derived growth factor --- p.46 / Chapter 2.3.9.4 --- Macrophage-colony stimulating factor --- p.47 / Chapter 2.3.9.5 --- Hepatocyte growth factor --- p.47 / Chapter 2.4 --- Molecular modulations in immune response --- p.47 / Chapter 2.4.1 --- NF-κB Signaling pathway --- p.47 / Chapter 2.4.2 --- NF-κB activation in endometriosis --- p.48 / Chapter 2.4.3 --- NF-κB activation in endometriosis associated peritoneal immune response --- p.50 / Chapter 2.5 --- The relationship between estrogen and immune response --- p.51 / Chapter 2.5.1 --- Effect of estrogen on immune cells and cytokines --- p.51 / Chapter 2.5.2 --- Effect of hormone treatment on immune response --- p.52 / Chapter 2.6 --- Summary --- p.53 / Chapter Chapter 3 --- Study Hypothesis and Objectives --- p.55 / Chapter 3.1 --- Hypothesis and aims --- p.56 / Chapter 3.2 --- Objectives --- p.56 / Chapter Chapter 4 --- Methodology --- p.57 / Chapter 4.1 --- Experimental design --- p.57 / Chapter 4.2 --- Cell-mediated immunity --- p.58 / Chapter 4.2.1 --- Experimental endometriosis in vivo --- p.58 / Chapter 4.2.2 --- Collection of peritoneal fluid, peritoneal cells and endometrial fragments --- p.59 / Chapter 4.2.3 --- Quantification of peritoneal immune cells --- p.59 / Chapter 4.3 --- MDSC functional analysis --- p.60 / Chapter 4.3.1 --- T cell suppression --- p.60 / Chapter 4.3.1.1 --- Collection and sorting of MDSC and T cells --- p.60 / Chapter 4.3.1.2 --- T cell proliferation assay --- p.61 / Chapter 4.3.2 --- Arginase production of MDSC --- p.62 / Chapter 4.3.2.1 --- Isolation of peritoneal G-MDSC and M-MDSC --- p.62 / Chapter 4.3.2.2 --- Arginase assay --- p.63 / Chapter 4.3.3 --- WrightGiemsa staining --- p.64 / Chapter 4.4 --- Kinetics of G-MDSC and M-MDSC --- p.64 / Chapter 4.4.1 --- Collection of peripheral mononuclear cells --- p.64 / Chapter 4.4.2 --- Collection of bone marrow cells --- p.65 / Chapter 4.5 --- MDSC depletion --- p.65 / Chapter 4.5.1 --- Interventions --- p.65 / Chapter 4.5.2 --- Sample collection --- p.66 / Chapter 4.5.3 --- BrdU staining --- p.66 / Chapter 4.6 --- Peritoneal cytokines and chemokines profile --- p.66 / Chapter 4.6.1 --- Cytokines antibody array --- p.66 / Chapter 4.6.2 --- Chemokines antibody array --- p.67 / Chapter 4.6.3 --- Data analysis of array --- p.68 / Chapter 4.6.4 --- ELISA validation --- p.69 / Chapter 4.6.5 --- Histology and immunohistochemistry --- p.69 / Chapter 4.6.5.1 --- Histology --- p.69 / Chapter 4.6.5.2 --- Immunohistochemistry --- p.70 / Chapter 4.7 --- MDSC proliferation and mobilization --- p.71 / Chapter 4.8 --- Statistics --- p.71 / Chapter Chapter 5 --- Results --- p.72 / Chapter 5.1 --- Histology of endometrial fragment --- p.72 / Chapter 5.2 --- Immune cell profiles --- p.72 / Chapter 5.3 --- Identification of MDSC --- p.73 / Chapter 5.3.1 --- T cell proliferation assay --- p.73 / Chapter 5.3.2 --- Arginase production --- p.74 / Chapter 5.3.3 --- Wright-Giemsa staining --- p.75 / Chapter 5.4 --- Kinetics of G-MDSC and M-MDSC --- p.75 / Chapter 5.5 --- MDSC depletion --- p.76 / Chapter 5.5.1 --- Gr-1 antibody depletion --- p.76 / Chapter 5.5.2 --- Size and weight of endometriotic lesions --- p.76 / Chapter 5.5.3 --- Growth and development of endometriotic lesions --- p.77 / Chapter 5.5.4 --- T cells and macrophages --- p.77 / Chapter 5.6 --- Cytokines and chemokines profiles --- p.78 / Chapter 5.6.1 --- Cytokine and chemokine profiles --- p.78 / Chapter 5.6.2 --- ELISA validation --- p.79 / Chapter 5.6.3 --- Cytokines expression in endometriotic lesions --- p.79 / Chapter 5.7 --- Proliferation and mobilization of MDSC by cytokine --- p.80 / Chapter Chapter 6 --- Discussion --- p.81 / Chapter 6.1 --- MDSC in early endometriosis --- p.81 / Chapter 6.1.1 --- Identification of MDSC --- p.81 / Chapter 6.1.1.1 --- Terminology of MDSC --- p.81 / Chapter 6.1.1.2 --- Subsets of MDSC --- p.81 / Chapter 6.1.1.3 --- Characteristics of MDSC --- p.82 / Chapter 6.1.2 --- MDSC in cancer --- p.83 / Chapter 6.1.2.1 --- Expansion and accumulation of MDSC in cancer --- p.83 / Chapter 6.1.2.2 --- Suppressive mechanisms of MDSC in cancer --- p.84 / Chapter 6.1.2.3 --- Suppressive mechanisms of MDSC subsets --- p.89 / Chapter 6.1.3 --- MDSC in inflammation --- p.90 / Chapter 6.1.4 --- MDSC in endometriosis --- p.90 / Chapter 6.2 --- Other immune cells in early endometriosis --- p.92 / Chapter 6.3 --- Cytokines and chemokines in early endometriosis --- p.93 / Chapter 6.4 --- Potentials of the study --- p.94 / Chapter 6.4.1 --- Potentials for new therapy --- p.95 / Chapter 6.4.1.1 --- Facilitating differentiation of MDSC --- p.95 / Chapter 6.4.1.2 --- Inhibiting MDSC proliferation --- p.96 / Chapter 6.4.1.3 --- Eliminating MDSC --- p.97 / Chapter 6.4.1.4 --- Blockade of MDSC suppressive function --- p.97 / Chapter 6.4.2 --- Potentials for new biomarkers --- p.97 / Chapter 6.5 --- Limitations of the study --- p.98 / Chapter 6.5.1 --- Lack of human study --- p.98 / Chapter 6.5.2 --- Lack of stable cell line --- p.98 / Chapter 6.5.3 --- Unknown mechanisms of MDSC in endometriosis --- p.99 / Chapter 6.5.4 --- Signaling pathway --- p.99 / Chapter Chapter 7 --- Conclusion and future plan --- p.100 / Reference --- p.102 / Chapter Appendices: --- Tables & Figures --- p.128

by Tai Fai Fok. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (p. 211-215). / Microfiche. Ann Arbor, Mich. UMI, 1998. 3 microfiches ; 11 x 15 cm.

Chemical hardness (eta), calculated by density functional theory (DFT), was firstly used as one of the chemical reactivity descriptors to set up the one descriptor 2D-QSAR model of platinum drugs. In this simple but promising model, the antitumour activities (log GI50) evaluated by National Cancer Institute (NCI) of structure-based groups containing normal sp 3 nitrogen and R,R-diamminecyclohexane (R,R-DACH) as the ligand showed good correlation. It was also demonstrated that silane and stereoisomers of DACH groups showed special patterns. This study also made use of the COMPARE program from NCI to evaluate the activity profile and the analysis of the data revealed these distinct patterns are influenced by the mechanism of the drugs. / Computer-aided drug design (CADD) techniques have been applied to establish quantitative structure-activity relationships (QSAR) and quantitative structure- property relationships (QSPR) models. Although these techniques are widely used in organic drugs, new metal-based drugs were hindered from development for lack of metal parameters, such as potent new platinum drugs as a major group of drugs used in cancer treatment. The purpose of the present study, therefore, is to generate novel platinum parameters based on previous work and then set up the simple QSAR/QSPR model with predictive abilities. / Finally, two 3D-QSAR and 3D-QSPR models obtained using Sybyl software. One was for demethylcantharidin (DMC) analogues as phosphatase 2A (PP2A) inhibitors. The other was describing the hydrophobicity of platinum drugs. In this research, the platinum atom was introduced to Sybyl and thus made it possible for the first time to use comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) methods to investigate platinum drugs. All 3D models indicated good predictive ability and thus provided an effective method to design new potent platinum drugs. / To clarify the pattern of stereoisomers of the DACH group, new platinum parameters was introduced to the AMBER software successfully. Moreover, stereoisomers of the DACH group which formed 1,2-GG intrastrand cross-links with DNA were studied by molecular dynamics (MD) simulations using AMBER. The calculated binding energies between R,R-DACH-Pt, S,S-DACH-Pt and cis-DACHPt moieties and DNA revealed a strong correlation with antitumour activities. The result provided more clues to understand the biological interactions of chiral platinum drugs. DNA structure analysis indicated that DNA tolerated the distortion resulted in the different Pt-DNA adducts and various local and global structure distortions were found. Natural bond orbital (NBO) analysis of hydrogen bonding on Pt-DNA adducts at a AGGC site revealed that R,R-DACH-Pt moiety alleviated the repulsion by unwinding the DNA, whereas the S,S-DACH-Pt adduct avoided the interaction by distorting the H bonds of binding site basepairs. Hence, the structural differences of chiral platinum drug led to its distinct activity. / Yang, Lifeng. / "June 2008." / Advisers: Steve C. F. Au Yeung; Yee-Ping Ho. / Source: Dissertation Abstracts International, Volume: 70-03, Section: B, page: 1541. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (p. 159-172). / 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.

Additionally, we showed that SeC induced S-phase arrest in MCF-7 cells associated with a marked decrease in the protein expression of cyclin A, D1 and D3 and cyclin-dependent kinases (CDK) 4 and 6, with concomitant induction of p21waf1/Cip1, p27Kip1 and p53. Expose of MCF-7 cells to SeC resulted in delayed onset of apoptosis as evidenced by caspase activation, PARP cleavage and DNA fragmentation. SeC treatment also triggered the activation of JNK, p38 MAPK, ERK and Akt phosphorylation. Inhibitors of ERK (U0126) or Akt (LY294002), but not JNK (SP600125) and p38 MAPK (SB203580), significantly suppressed SeC-induced S-phase arrest and apoptosis in MCF-7 cells. In conclusion, our findings establish a mechanistic link between the PI3K/Akt pathway, MAPK pathway and SeC-induced cell cycle arrest and apoptosis in human breast cancer cells. (Abstract shortened by UMI.) / The role of selenium as potential cancer chemopreventive and chemotherapeutic agents has been supported by epidemiological, preclinical and clinical studies. Although cell apoptosis has been evidenced as a critical mechanism mediating the anticancer activity of selenium, the underlying molecular mechanisms remain elusive. In the present study, selenocystine (SeC), a novel organic selenocompound, is identified as a novel antiproliferative agent with a broad spectrum of inhibition against eight human cancer cell lines with the IC50 values ranged from 3.6 to 37.0 muM. Despite this potency, SeC was relatively nontoxic toward HS68 human fibroblasts with an IC 50 value exceeded 400 muM. Further investigation on the molecular mechanisms indicated that SeC induced caspase-independent apoptosis in MCF-7 breast carcinoma cells, which was accompanied by poly(ADP-ribose) polymerase (PARP) cleavage, caspase activation, DNA fragmentation, phosphatidylserine exposure and nuclear condensation. Moreover, SeC induced the loss of mitochondrial membrane potential (DeltaPsim) by regulating the expression and phosphorylation of pro-surivival and pro-apoptotic Bcl-2 family members. Loss of DeltaPsim led to the mitochondrial release of cytochrome c and apoptosis-inducing factor (AIF) which subsequently translocated into the nucleus and induced chromatin condensation and DNA fragmentation. MCF-7 cells exposed to SeC shown increase in total p53 and phosphorylated p53 on serine residues of Ser15, Ser20, and Ser392 prior to mitochondrial dysfunction. Silencing and attenuation of p53 expression with RNA interference and pifithrin-alpha treatment respectively, partially suppressed SeC-induced cell apoptosis. Furthermore, generation of reactive oxygen species (ROS) and subsequent induction of DNA strand breaks were found to be upstream cellular events induced by SeC. The thiol-reducing antioxidants, N-acetylcysteine and glutathione, completely blocked the initiation and execution of cell apoptosis. Taken together, these results suggest that SeC, as a promising anticancer selenocompound, induces caspase-independent apoptosis in MCF-7 cells mediated by ROS generation and p53 phosphorylation through regulating the mitochondrial membrane permeability. / Chen, Tianfeng. / Adviser: Yun-Shing Wong. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3260. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 124-136). / 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.

Bioactivity-guided isolation of SFE oil led to the identification of another sesquiterpene lactone, 6-O-angeloylprenolin, containing the bioactive alpha, beta-unsaturated cyclopentenone. MTT results showed that CNE cells were more susceptible to 6-O-angeloylenolin than the normal Hs68 cells. Besides, the inhibitory effect of 6-O -angeloylenolin on the CNE cells was slightly stronger than that of cisplatin, the positive control, albeit statistical insignificance. / Both volatile oils prepared by supercritical fluid extraction (SFE) and steam distillation (SD) were evaluated for their anti-NPC potential. Results showed that SFE oil was much stronger than that of SD oil. SFE oil significantly inhibited the growth of CNE cells by dysfunctioning the mitochondria and activating caspases. Gas chromatography-mass spectrometry analysis revealed that the responsible principals in the SFE oil were likely homologues of sesquiterpene lactones. / Centipeda minima (L.) A. Br. (Compositae), a Chinese medicinal herb, is used to treat nasopharyngeal carcinoma (NPC) in the Chinese folk. However, there is a paucity of information on its anticancer activities. In particular, both of its anti-NPC potential and the potent constituents remain elusive. / In this study, the n-hexane fraction of C. minima showed broad spectrum of inhibitory effects on five human cancer cell lines, including the breast carcinoma MCF7 cells, the prostate carcinoma PC-3 cells, the hepatocellular carcinoma Hep G2 cells, the nasopharyngeal cancer CNE cells and the acute promyelocytic leukemia HL-60 cells, with IC 50 values ranging from 6.1 to 47.3 mug/mL. Bioactivity-guided separation of the n-hexane fraction using the CNE cells as the cellular system led to the isolation of a sesquiterpene lactone, 2beta-(isobutyryloxy)florilenalin (IF), which contained the bioactive alpha-methylene-gamma-lactone ring. IF significantly induced CNE cell death with an IC50 value of 3.1 mug/mL. Despite this potency, its effect on the normal Hs68 cells was much weaker, with an IC50 value larger than 50 mug/mL. Its inhibitory effect on the CNE cells ascribed to apoptotic induction as evidenced by the cumulation of sub-G1 cell population, DNA fragmentation and nuclear condensation, caspase-3 activation and poly (ADP-ribose) polymerase (PARP) cleavage. Mechanistic study showed that both extrinsic and intrinsic apoptotic pathways were activated. In the extrinsic pathway, IF activated caspase-8, which further induced the activation of caspase-3 and caspase-7. In the intrinsic pathway, IF regulated the expressions of Bcl-2 family proteins, followed by depletion of mitochondrial membrane potential (Delta&PSgr;m), the release of cytochrome c to cytosol, the activation of caspase-9 and other downstream caspases, and finally the induction of apoptosis. / Mechanistic investigation showed that 6-O-angeloylenolin caused cell cycle arrest at S and G2/M phases and induced apoptosis in CNE cells. For the cell cycle arrest, a sharp decrease was found in the expressions of cyclin D1, cyclin D3, cdc25c, and p-cdc25c, with concomitant decrease in CDK4, cyclin A, cyclin E, p-Rb(Ser780), p21Waf1/Cip1, cdc2 and p-cdc2. For the induction of apoptosis, externalization of phosphatidylserine and depletion of Delta&PSgr;m prior to the detection of sub-G1 peak were found. Other apoptotic features including the presence of apoptotic bodies, the activation of caspase-3 activity and the cleavage of PARP were observed. Activation of caspase-8 and caspase-10 was detected. Besides, 6-O -angeloylenolin induced the release of cytochrome c and AIF to cytosol. The former formed apoptosome with caspase-9, further activated the downstream caspase-3 and caspase-7 and cleaved PARP, while the latter was translocated into the nucleus and caused large-scale DNA fragmentation. Failure of the pan-caspase inhibitor, z-VAD-fmk, to interrupt the apoptotic induction by 6-O-angeloylenolin suggested that caspase-independent pathway was involved. 6-O-Angeloylenolin was able to activate Akt, ERK and JNK pathways. But only with the addition of JNK inhibitor (SP600125), significant suppression of the 6-O-angeloylenolin-induced apoptosis was observed, suggesting the involvement of the JNK pathway in the apoptotic pathway. Taken together, this study provided a better mechanistic insight into the potential application of 6-O-angeloylenolin as a candidate for NPC treatment. / Overall, this study revealed that two sesquiterpene lactones, including IF and 6-O-angeloylenolin were found to be responsible for the potent anti-NPC effect of C. minima. This study reiterates the notion that Chinese medicinal herbs traditionally applied to cancer treatment may be good sources of anticancer drug discovery, and sesquiterpene lactone may be a group of noteworthy lead compounds displaying anti-NPC potential. / Su, Miaoxian. / Adviser: Hau Yin Chung. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 100-113). / 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.

Hong, Sijia. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 197-234). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

A breast cancer cell line stably transfected with the CYP19 gene had been employed for aromatase inhibition. Among the phytochemicals tested, the major dietary flavonoids, such as genistein and daidzein, produced very weak inhibition. On the other hand, the red clover isoflavone biochanin A, the hydroxychalcone butein and the red grape phytoalexin resveratrol were found to be effective aromatase inhibitors. Cell proliferation assay had shown that they could inhibit ER-positive cell proliferation induced by testosterone, and the inhibitory effect was specifically attributed to the reduction of estrogen synthesis. In another breast cancer cell line SK-BR-3, resveratrol, biochanin A and genistein inhibited CYP19 both in enzyme and promoter I.3/II transcriptional levels. The element responsible for the inhibition of aromatase by these phytoestrogens should fall within the region between -556 to -446 by upstream of exon II. / Breast cancer is one of the most common cancers affecting women. Estrogen plays an important role in breast cancer initiation and development. The majority of breast tumors are initially dependent upon estrogen to support their growth. Most breast cancers occur in the postmenopausal period. However, the intra-tumoral estradiol (E2) is maintained at a high level equivalent to the pre-menopausal status. High intra-tumoral E2 level in postmenopausal women is sustained by the biosynthesis of estrogens in the tumorous tissue. / Genistein and Biochanin A, ranged from 0.1 to 10 muM, might act as estrogen agonist and induced aromatase activity and promoter I.1 transactivation in ERalpha-transfected SK-BR-3 cells. (Abstract shortened by UMI.) / The aromatase enzyme, CYP19, belongs to a family of P450 enzyme. As a final rate-limiting step in estrogen biosynthesis, it catalyzes the conversion of C 19 steroids to estrogens. The expression of CYP19 is tissue-specific, and is regulated by alternate promoter usage. The use of aromatase inhibitors for breast cancer treatment has become a major therapeutic approach. / The consumption of some phytochemicals protects against breast cancer. Yet the mechanisms are far from clear. In my present study, various phytochemicals, including phytoestrogens, monoterpenes and carotenoids, were evaluated for their effect on aromatase. / Wang Yun. / "July 2005." / Adviser: Lai-Kwok Leung. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3716. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 145-169). / 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. / Abstract in English and Chinese. / School code: 1307.

Colorectal cancer is the second leading cause of cancer-related mortality in Hong Kong and lack of selectivity has limited the success of conventional chemotherapy. Given the recent interest in the anti-cancer effects of traditional Chinese medicine (TCM), there are two approaches to studying its bioactivity: as a mixture of ingredients or as single compounds. The objective of the present study is to examine the anti-tumor effects of Angelicae Sinensis Radix (DG) and Garcinia hanburyi resin (TH) using both approaches, respectively, as they are traditionally used to treat inflammation. In the present study, their anti-cancer effects and the mechanisms of actions were examined for the development of potential novel chemotherapeutic drugs for colon cancer since inflammation is a predisposing factor for colon cancer. / DG extract and its three main bioactive phtbalides: n-butylidenephthalide, senkyunolide A and z-ligustilide (LGT), were found to be cytotoxic to HT-29 cells with IC50 values (24 h) of 20.70 +/- 0.85, 72.51 +/- 8.65, 18.74 +/- 1.14 and 41.98 +/- 3.64 mug/ml, respectively. The results evidenced that LGT induced G0/G 1 arrest and apoptosis, triggering cleavage of PARP, pro-caspases-3, -8 and -9 and nuclear fragmentation. LGT and cisplatin synergistically reduced the viability of HT-29 cells. More interestingly, DG extract was more potent than individual phthalides, suggesting that there are other bioactive components and/or synergistic interactions. / Individual compounds purified from TH were investigated because gambogic acid isolated from this herb has been used clinically to treat cancer, 30-Epicambogin (EPC) and guttiferone K (GUTK) showed the highest cytotoxic selectivity and potency on HT-29 cells among 15 isolated compounds. IC50 values (24 h) for EPC and GUTK in HT-29 cells were 5.36+/-0.25 and 5.39+/-0.22 muM, respectively, and both induced G0/G1 arrest by down-regulation of cyclins D1, D3, CDK4 and CDK6, while up-regulation of p21Waf1/CiP1 and p27KiP1. Both compounds triggered the activation of caspases-3, -8 and -9 in apoptosis. The in vivo anti-tumor effects of GUTK were further investigated by using a subcutaneous Colon-26 mouse tumor model. GUTK (10 mg/kg i.p.) reduced tumor volume by 33.6% and potentiated the anti-tumor effects of 5-fluorouracil when administered concurrently. / Our findings revealed that DG rather than individual phthalides, is worthy for further study as a potential anti-cancer drug, due to the synergistic interactions among multi-components in the herb. On the other hand, EPC and GUTK, isolated from TH have potential to be developed as novel anti-tumor candidates for combination use with 5-fluorouracil. The results strongly support the use of different approaches to study TCM for chemotherapy, according to its traditional and empirical use. / Subsequently, the anti-proliferative effects of DG and Chuanxiong Rhizoma (CX) extracts and mixtures containing three phthalides in the proportions similar to their presence in both extracts were examined, since CX also contains the same phthalides, but in different proportions. DG extract was significantly more potent than its corresponding phthalide mixture to inhibit cancer cell proliferation and synergistic interaction was observed among the phthalides and other bioactive components, while the phthalides in CX extract interacted antagonistically with other components. / Kan, Lai Ting Winnie. / Adviser: Ge Lin. / Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 267-311). / 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.