Sze, Siu Ching. / Thesis M.Phil. Chinese University of Hong Kong 2015. / Includes bibliographical references (leaves 109-124). / Abstracts also in Chinese. / Title from PDF title page (viewed on 24, October, 2016).

He, Mian. / Thesis Ph.D. Chinese University of Hong Kong 2015. / Includes bibliographical references (leaves 165-188). / Abstracts also in Chinese. / Title from PDF title page (viewed on 24, October, 2016).

Orientador: Cecilia Amelia Fazzio Escanhoela / Dissertação (mestrado) - Universidade Estadual de Campínas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-15T12:26:43Z (GMT). No. of bitstreams: 1 Segatelli_Vanderlei_M.pdf: 1173808 bytes, checksum: efbd82ac6962165e42a166dfef483327 (MD5) Previous issue date: 2010 / Resumo: O carcinoma hepatocelular (CHC) destaca-se como uma das formas mais comuns de câncer no mundo, correspondendo a aproximadamente 90% das neoplasias primárias do fígado, sendo a cirrose seu mais importante fator de risco. A sua progressão, no entanto, depende de inúmeros fatores e, entre estes, a angiogênese assume papel fundamental, principalmente por ser o CHC um tumor altamente vascularizado. É sabido que tumores sólidos, ao atingirem volume de 1 a 2mm3, passam a ter seu crescimento dependente de neovascularização e no fígado esta se faz através de capilarização sinusoidal, processo pelo qual ocorrem alterações morfológicas nos sinusóides hepáticos, com perda das fenestrações e deposição de membrana basal. Neste estudo retrospectivo avaliamos a angiogênese, através dos marcadores imuno-histoquímicos CD34 e CD105 (Endoglina), com determinação da densidade microvascular (DMV) em 44 produtos de hepatectomia total, compreendendo 44 nódulos neoplásicos (CHC), 44 nódulos regenerativos (NR) e 15 nódulos com características microscópicas de lesões pré-malignas (ou Nódulos Displásicos - ND), sendo 8 de baixo grau e 7 de alto grau. A avaliação compreendeu a determinação da DMV no centro e na periferia de todos os nódulos. Para análise estatística foi utilizada análise descritiva com apresentação de medidas de posição e dispersão para variáveis contínuas; para a comparação dos grupos foi utilizada a ANOVA, considerando-se p<0,05 estatisticamente significante. O CD34 demonstrou positividade difusa no CHC, com valores médios da DMV de 38,3±12,6 e 34,2±11,2 no centro e periferia, respectivamente. Já o CD105 apresentou expressão sinusoidal predominantemente periférica no CHC, com valores médios da DMV de 6,2±4,1 e 10,7±4,4, com diferenças significativas entre os grupos (p<0,0001). Também foram observadas diferenças na imunoexpressão entre os marcadores em NR (p<0,0001) e ND (p<0,0019). Concluímos que existem diferenças na imunoexpressão do CD34 e CD105 entre os nódulos benignos e malignos, com gradual aumento da neovascularização, bem caracterizado pelo CD34, devido ao processo de capilarização sinusoidal. O CD105, expresso em "subset" de microvasos neoformados, pode ser um indicador de diferenciação diagnóstica entre NR e CHC / Abstract: Hepatocellular carcinoma (HCC) is one of the most common forms of cancer worldwide, accounting for about 90% of primary tumors of the liver. Cirrhosis is the most important risk factor for HCC. However, the progression of hepatocellular carcinoma is dependent on a number of factors. Among these factors, angiogenesis plays a major role, mainly because HCC is a highly vascular tumor. It is known that when solid tumors reach a volume of 1 to 2mm3 their growth depends on neovascularization. In the liver, this occurs through sinusoidal capillarization, a process by which morphologic alterations occur in the hepatic sinusoids, with loss of fenestrations and basement membrane depositions. In this retrospective study, we evaluated angiogenesis by using immunohistochemical markers CD34 and CD105 (Endoglin), with determination of tumor microvessel density (MVD) in 44 products of total hepatectomy, including 44 malignant nodules (HCC), 44 regenerative nodules (RN) and 15 nodules with microscopic characteristics of preneoplastic lesions (or Dysplastic Nodules-DN), 8 being low-grade and 7 high-grade. The evaluation encompassed measurement of MVD in the center and periphery of all nodules. For statistical analysis, a descriptive analysis with measurements of position and dispersion for continuous variables was used. ANOVA was used for group comparison, considering p<0.05 as statistically significant. CD34 demonstrated a diffuse positivity in HCC, with median MVD values of 38.3±12.6 and 34.2±11.2 in the center and periphery, respectively. On the other hand, CD105 exhibited sinusoidal expression predominantly in the periphery of the HCC, with median MVD values of 6.2±4.1 and 10.7±4.4, with significant differences between groups (p<0.0001). Differences in the immunoexpression between markers in RN (p<0.0001) and DN (p<0.0019) were also observed. In conclusion, there are differences in the immunoexpression of CD34 and CD105 between benign and malignant nodules, with a gradual increase in neovessel formation, well-characterized by CD34, due to a process of sinusoidal capillarization. CD105, expressed in a "subset" of newly formed microvessels may be an indicator of the diagnostic differentiation between RN and HCC / Mestrado / Anatomia Patologica / Mestre em Ciências Médicas

Lee Siu-wah. / Thesis submitted in: December 1999. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves [106]-116). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Figures --- p.ix / List of Tables --- p.x / Abbreviations --- p.xi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Hepatocellular Carcinoma (HCC) --- p.2 / Chapter 1.2 --- Etiology of Hepatocellular Carcinoma --- p.5 / Chapter 1.2.1 --- Viral Infection --- p.5 / Chapter 1.2.1.1 --- Hepatitis B Virus --- p.5 / Chapter 1.2.1.2 --- Hepatitis C Virus --- p.7 / Chapter 1.2.2 --- Cirrhosis and Chronic Inflammation --- p.8 / Chapter 1.2.3 --- Aflatoxin --- p.9 / Chapter 1.3 --- Genetic Studies of Hepatocellular Carcinoma --- p.9 / Chapter 1.3.1 --- Conventional Cytogenetics --- p.9 / Chapter 1.3.2 --- Molecular Cytogenetics --- p.12 / Chapter 1.3.3 --- Molecular Genetic Studies --- p.12 / Chapter 1.3.3.1 --- Proto-oncogenes --- p.12 / Chapter 1.3.3.2 --- Tumour Suppressor Genes --- p.13 / Chapter 1.3.3.3 --- Cell Cycle Genes --- p.14 / Chapter 1.4 --- Background of Study --- p.16 / Chapter 1.5 --- Objectives of Study --- p.17 / Chapter Chapter 2 --- Material and Methods --- p.18 / Chapter 2.1 --- Materials --- p.19 / Chapter 2.2 --- Analysis of Chromosomal Imbalances by Comparative Genomic Hybridization --- p.23 / Chapter 2.2.1 --- Comparative Genomic Hybridization --- p.23 / Chapter 2.2.2 --- Methods / Chapter 2.2.2.1 --- Preparation of Normal Metaphases --- p.30 / Chapter 2.2.2.2 --- Extraction of High Molecular Weight DNA --- p.30 / Chapter 2.2.2.3 --- Labeling of DNA by Nick Translation --- p.31 / Chapter 2.2.2.4 --- Labeling Efficiency --- p.31 / Chapter 2.2.2.5 --- Preparation of Probe --- p.33 / Chapter 2.2.2.6 --- Hybridization --- p.33 / Chapter 2.2.2.7 --- Washing and Detection of Signals --- p.35 / Chapter 2.2.2.8 --- Image Acquisition and Analysis --- p.35 / Chapter 2.2.2.9 --- Control Experiments --- p.36 / Chapter 2.3 --- Positional Mapping of Novel Amplicon by Interphase Cytogenetics --- p.39 / Chapter 2.3.1 --- Fluorescence in situ Hybridization --- p.39 / Chapter 2.3.2 --- Using Yeast Artificial Chromosomes (YAC) as Probe --- p.41 / Chapter 2.3.3 --- Methods --- p.48 / Chapter 2.3.3.1 --- Culture of Yeast Artificial Chromosomes --- p.48 / Chapter 2.3.3.2 --- Extraction of Total YAC DNA --- p.48 / Chapter 2.3.3.3 --- Verification of YAC Clones for Chimerism by FISH --- p.49 / Chapter 2.3.3.4 --- Inter-Alu-PCR --- p.49 / Chapter Chapter 3 --- Assessment of Genetic Changes in HCC by Comparative Genomic Hybridization (CGH) --- p.57 / Chapter 3.1 --- Introduction --- p.58 / Chapter 3.2 --- Materials and Methods --- p.58 / Chapter 3.2.1 --- Patients and Specimens --- p.58 / Chapter 3.2.2 --- Comparative Genomic Hybridization --- p.60 / Chapter 3.2.3 --- Statistical Analysis --- p.60 / Chapter 3.3 --- Results --- p.61 / Chapter 3.3.1 --- Overall Copy Number Aberrations in 67 HCC and Surrounding Cirrhotic Tissues --- p.61 / Chapter 3.3.2 --- TNM Staging --- p.61 / Chapter 3.3.3 --- Tumour Size --- p.72 / Chapter 3.3.4 --- Serum AFP Elevation --- p.72 / Chapter 3.3.5 --- Chromosomal Aberrations in HCC arising from Cirrhotic and Non- cirrhotic Livers --- p.72 / Chapter 3.4 --- Discussion --- p.73 / Chapter 3.4.1 --- Recurrent Gains --- p.73 / Chapter 3.4.2 --- Recurrent Losses --- p.75 / Chapter 3.4.3 --- Tumour Progression --- p.76 / Chapter 3.5 --- Conclusion --- p.78 / Chapter Chapter 4 --- Positional Mapping of a Novel Amplicon on Chromosome 1q21-q25 by Interphase Cytogenetics --- p.79 / Chapter 4.1 --- Introduction --- p.80 / Chapter 4.2 --- Materials --- p.82 / Chapter 4.3 --- Methods --- p.82 / Chapter 4.3.1 --- Preparation of Paraffin-embedded Tissue Sections --- p.82 / Chapter 4.3.2 --- Verification of YAC Probes for Chimerism --- p.83 / Chapter 4.3.3 --- Hybridization Efficiency of Test and Reference Probes --- p.83 / Chapter 4.3.4 --- Slide Pretreatment and FISH with YAC Probes --- p.88 / Chapter 4.3.5 --- Scoring of FISH Signals --- p.88 / Chapter 4.4 --- Results --- p.89 / Chapter 4.4.1 --- Relative Copy Number Gain --- p.89 / Chapter 4.4.2 --- Intratumour Heterogeneity --- p.90 / Chapter 4.5 --- Discussion --- p.90 / Chapter 4.6 --- Further Studies --- p.104 / Chapter 4.6.1 --- Fine Mapping of Chromosomal Region 1 q21 --- p.104 / Chapter 4.6.2 --- Isolation of Novel Genes in the Amplicon --- p.105 / Chapter 4.6.3 --- Expression Status of the EDC Genes --- p.105 / References --- p.106

Cheung Shiu-fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 131-139). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.iii / 摘要 --- p.vi / TABLE OF CONTENTS --- p.ix / LIST OF FIGURES --- p.xvi / LIST OF TABLES --- p.xviii / ABBREVIATIONS --- p.xix / PUBLICATION --- p.xxi / AWARD --- p.xix / Chapter SECTION 1. --- INTRODUCTION OF HEPATOCELLULAR CARCINOMA --- p.1 / Chapter 1.1 --- Epidemiology of Hepatocellular Carcinoma --- p.1 / Chapter 1.2 --- Etiologies of HCC --- p.1 / Chapter 1.2.1 --- Hepatitis B Virus and Hepatitis C Virus --- p.2 / Chapter 1.2.2 --- Aflatoxins and Alcohol --- p.3 / Chapter 1.3 --- Major Diagnostic and Prognostic Markers of HCC --- p.4 / Chapter 1.3.1 --- Biochemical Tumor Markers --- p.5 / Chapter 1.3.2 --- Clinico-pathological Features of HCC --- p.6 / Chapter SECTION 2. --- THE MITOCHONDRION --- p.7 / Chapter 2.1 --- Structure of the Mitochondrial Genome --- p.9 / Chapter 2.1.1 --- Nicotinamide Adenine Dinucleotide Dehydrogenase --- p.10 / Chapter 2.1.2 --- Cytochrome b --- p.10 / Chapter 2.1.3 --- Cytochrome c Oxidase --- p.11 / Chapter 2.1.4 --- ATP Synthase --- p.11 / Chapter 2.1.5 --- Ribosomal RNA --- p.11 / Chapter 2.1.6 --- Transfer RNA --- p.12 / Chapter 2.1.7 --- Displacement Loop --- p.12 / Chapter 2.2 --- Replication of Mitochondrial DNA --- p.17 / Chapter 2.3 --- Transcription of Mitochondrial DNA --- p.17 / Chapter SECTION 3. --- PHYSIOLOGY OF MITOCHONDRIA --- p.19 / Chapter 3.1 --- Energy Production by Oxidative Phosphorylation (OXPHOS) --- p.19 / Chapter 3.2 --- Programmed Cell Death: Apoptosis --- p.22 / Chapter 3.3 --- Morphology of Mitochondria in Hepatocytes --- p.25 / Chapter SECTION 4. --- MUTATIONS OF MITOCHONDRIAL DNA --- p.26 / Chapter 4.1 --- Special Terms Used in This Study --- p.26 / Chapter 4.1.1 --- Somatic Mutations and Polymorphisms --- p.26 / Chapter 4.1.2 --- Homoplasmic and Heteroplasmic Mutations --- p.26 / Chapter 4.2 --- Factors Causing High Mutation Frequency in mtDNA --- p.27 / Chapter 4.2.1 --- Presence of Reactive Oxygen Species --- p.27 / Chapter 4.2.2 --- Lack of Protective Histories --- p.28 / Chapter 4.2.3 --- Limited DNA Repair Mechanism --- p.29 / Chapter 4.3 --- Theories of Homoplasmic Mutations --- p.31 / Chapter 4.3.1 --- Replicative Advantage on Mutated mtDNA Sequence Selection --- p.31 / Chapter 4.3.2 --- Random Mutagenesis and Segregation --- p.32 / Chapter 4.4 --- MtDNA Mutations in Mitochondrial Disease and Aging --- p.33 / Chapter 4.5 --- MtDNA Deletions in Cancer --- p.34 / Chapter 4.6 --- Somatic Mutations of MtDNA in Various Cancers --- p.35 / Chapter 4.6.1 --- Frequencies of Somatic Mutations --- p.35 / Chapter 4.6.2 --- Distribution of Somatic Mutations in mtDNA --- p.36 / Chapter 4.7 --- Somatic Mutations of Mitochondrial DNA in HCC --- p.37 / Chapter SECTION 5. --- OBJECTIVES OF THIS STUDY --- p.44 / Chapter SECTION 6. --- MATERIALS AND METHODS --- p.46 / Chapter 6.1 --- Patients and Samples Collection --- p.46 / Chapter 6.2 --- DNA Extraction from Liver Tissues --- p.46 / Chapter 6.3 --- Amplification of Mitochondrial DNA by Polymerase Chain Reaction --- p.51 / Chapter 6.3.1 --- Design of Primers --- p.51 / Chapter 6.3.2 --- PCR Conditions and Contents --- p.54 / Chapter 6.3.3 --- Assessment of PCR Products by Agarose Gel Electrophoresis --- p.54 / Chapter 6.4 --- Purification of PCR Products --- p.54 / Chapter 6.5 --- Cyclesequencing of Mitochondrial DNA --- p.55 / Chapter 6.5.1 --- Design of Primers --- p.55 / Chapter 6.5.2 --- PCR Contents and Cycle Sequencing Procedures --- p.56 / Chapter 6.6 --- Purification of Sequencing Products --- p.56 / Chapter 6.7 --- Sequence Analysis by Automated Sequencer --- p.57 / Chapter 6.7.1 --- Preparation of Polyacrylamide Gel --- p.57 / Chapter 6.7.2 --- Sequence Analysis by Automated Sequencer --- p.58 / Chapter 6.7.3 --- "Search for Sequence Variants, Polymorphisms and Somatic Mutations" --- p.58 / Chapter 6.8 --- Further Studies on mtDNA Mutations --- p.59 / Chapter 6.8.1 --- Sequence Analysis in Buffy Coat --- p.59 / Chapter 6.8.2 --- Detection of the Presence of Somatic mtDNA Mutations in Plasma --- p.59 / Chapter 6.8.3 --- Frequency of Mutations in Two Nucleotide Repeat Sequences --- p.60 / Chapter 6.9 --- Clinical Data and Statistical Analysis --- p.61 / Chapter 6.9.1 --- Clinical and Pathological Data --- p.61 / Chapter 6.9.2 --- Statistical Analysis --- p.61 / Chapter SECTION 7. --- RESULTS --- p.63 / Chapter 7.1 --- Sequence Analysis of the Entire Mitochondrial Genome --- p.63 / Chapter 7.1.1 --- Sequence Variants and Polymorphisms --- p.63 / Chapter 7.1.2 --- Somatic Mutations --- p.71 / Chapter 7.2 --- Study of Mitochondrial Sequence in Lymphocytes --- p.78 / Chapter 7.3 --- Detection of Tumor DNA in Serum --- p.78 / Chapter 7.4 --- Analysis of Nucleotide Repeat Sequences --- p.79 / Chapter 7.4.1 --- General Results --- p.79 / Chapter 7.4.2 --- Statistical Analysis --- p.84 / Chapter SECTION 8. --- DISCUSSION --- p.89 / Chapter 8.1 --- Comparative Analysis of mtDNA Mutations with Two Previous HCC Studies --- p.89 / Chapter 8.1.1 --- Number of Cases and Region Studied --- p.89 / Chapter 8.1.2 --- Number and Distribution of Mutations in Normal Controls --- p.89 / Chapter 8.1.3 --- Number of Somatic Mutations --- p.90 / Chapter 8.1.4 --- Distribution of Somatic Mutations --- p.91 / Chapter 8.2 --- Similarities of Somatic mtDNA Mutations in This Study with Other Cancer Types --- p.93 / Chapter 8.2.1 --- Frequency and Distribution of Somatic Mutations --- p.93 / Chapter 8.2.2 --- Number of Homoplasmic Mutations --- p.93 / Chapter 8.3 --- Evaluation of Somatic Mutations of mtDNA in This Study --- p.96 / Chapter 8.3.1 --- Specificity of Somatic Mutations in Tumor Proved by Sequence Analysis in Lymphocytes --- p.96 / Chapter 8.3.2 --- Importance of Conserved Amino Acid Sequences with Other Species to the Presence of Somatic Mutations in Tumor --- p.96 / Chapter 8.3.3 --- Four Somatic Mutation Sites Are Detected in More than One Cancer Type --- p.101 / Chapter 8.3.4 --- Presence of Homoplasmic and Heteroplasmic Mutations --- p.101 / Chapter 8.3.5 --- Absence of Large-scale Deletions in Tumor Tissues --- p.102 / Chapter 8.4 --- Mutation Hotspots Region: Hypervariable Displacement-loop --- p.103 / Chapter 8.5 --- D310 Mononucleotide Repeats --- p.106 / Chapter 8.5.1 --- Description of D310 Mononucleotide Repeats --- p.106 / Chapter 8.5.2 --- Possible Causes of Varied Sequences at D310 --- p.106 / Chapter 8.5.3 --- Appearance of Nucleotide Repeats at D310 in Tumors --- p.107 / Chapter 8.5.4 --- Possible Outcomes of D310 Aberrations in mtDNA Replication and Transcription --- p.108 / Chapter 8.5.5 --- Comparison of D310 Alternations in HCC with Other Cancers --- p.109 / Chapter 8.6 --- Other Nucleotide Repeat Sequences --- p.112 / Chapter 8.6.1 --- The CA Dinucleotide Repeats --- p.112 / Chapter 8.6.2 --- Other Nucleotide Repeat Sequences Showing Genome Instability --- p.112 / Chapter 8.7 --- Evaluation of Somatic mtDNA Mutations as a Cancer Diagnostic Marker --- p.114 / Chapter 8.7.1 --- Coding Region --- p.114 / Chapter 8.7.2 --- D-loop Region --- p.114 / Chapter 8.7.3 --- D310 Nucleotide Repeats --- p.115 / Chapter 8.7.4 --- Possibility of Detecting Somatic Mutations in Serum --- p.116 / Chapter 8.8 --- Somatic mtDNA Mutations May Be a Prognostic Marker in HCC --- p.117 / Chapter 8.8.1 --- Possible Problems in Current Prognostic Factors --- p.117 / Chapter 8.8.2 --- Interpretation of Results --- p.117 / Chapter 8.8.3 --- Prognostic Values of Somatic Mutations at D310 --- p.118 / Chapter 8.9 --- Hypothesis of Somatic MtDNA Mutations on Tumorigenesis and Tumor Progression --- p.119 / Chapter 8.9.1 --- Somatic mtDNA Mutations Decline OXPHOS and May Inactivate Apoptotic Pathways --- p.119 / Chapter 8.9.2 --- Moderate Reactive Oxygen Species Production May Promote Mitosis --- p.120 / Chapter 8.10 --- Possible Appearance of Somatic Mutations in HCC with Chronic HBV Infection --- p.123 / Chapter 8.11 --- Possibility of HBx Protein Integration to MtDNA Mutations --- p.123 / Chapter 8.12 --- Conclusions --- p.125 / Chapter SECTION 9. --- LIMITATIONS AND FURTHER STUDIES --- p.127 / Chapter 9.1 --- Limitations and Improvements of Study --- p.127 / Chapter 9.1.1 --- Small Sample Size --- p.127 / Chapter 9.1.2 --- Sequence Analysis Method --- p.127 / Chapter 9.1.3 --- Fidelity of PCR Reactions and Long-range PCR Fragments --- p.128 / Chapter 9.2 --- Further Studies --- p.129 / Chapter SECTION 10. --- REFERENCES --- p.131

Lam Wai-Chun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves [64]-[73]). / Abstracts in English and Chinese. / Abstract (in English) --- p.i-ii / Abstract (in Chinese) --- p.iii -iv / Acknowledgements --- p.v / Table of contents --- p.vi -ix / List of Figures --- p.x / List of Tables --- p.x / Abbreviations --- p.xi -xii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Hepatocellular Carcinoma (HCC) --- p.1-2 / Chapter 1.2 --- Major risk factors of HCC / Chapter (1) --- Hepatitis B Virus (HBV) --- p.2-4 / Chapter (2) --- Hepatitis C Virus (HCV) --- p.5-6 / Chapter (3) --- Cirrhosis --- p.6 / Chapter (4) --- Dietary alfatoxin B1 (AFB1) --- p.6 -7 / Chapter (5) --- Alcoholic consumption --- p.7 / Chapter (6) --- Iron overload --- p.8 / Chapter 1.3 --- Genetic aberrations in HCC --- p.8-9 / Chapter (1) --- Chromosomal loss --- p.10-13 / Chapter (2) --- Chromosomal gains --- p.13-15 / Chapter 1.4 --- roposed study --- p.15 / Chapter (1) --- Hypomethylation of heterochromatin in chromosome 1q copy number gain. --- p.16 / Chapter (2) --- ositional mapping on 1q21 - q22 by interphase cytogenetics. --- p.16-17 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Southern Blot Analysis for Satellite DNA Hypomethylation. --- p.18-19 / Chapter 2.1.2 --- ositional Mapping by Interphase Cytogenetics. --- p.19 -24 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Southern Blot Analysis for Satellite DNA Hypomethylation / Chapter (1) --- Extraction of high molecular weight DNA --- p.25 / Chapter (2) --- DNA digestion with methyl-sensitive restriction enzyme --- p.25 -26 / Chapter (3) --- Control for the complete DNA digestion. --- p.26 / Chapter (4) --- Southern Blotting. --- p.26 -27 / Chapter 2.2.2 --- ositional Mapping by Interphase Cytogenetics / Chapter (1) --- Yeast Artificial Chromosome (YAC) --- p.28 -29 / Chapter (i) --- YAC culturing --- p.29 -30 / Chapter (ii) --- YAC DNA extraction --- p.30 -31 / Chapter (iii) --- Inter-Alu-Polymerase Chain Reaction --- p.32 -33 / Chapter (2) --- -1 derived Bacterial Artificial Chromosome (PAC) --- p.34 / Chapter (i) --- AC culturing and DNA extraction --- p.34 -35 / Chapter (3) --- FISHrobe labeling by nick translation. --- p.35 / Chapter (4) --- FISHrobereparation --- p.36 / Chapter (5) --- Dot-blot analysis. --- p.36 -37 / Chapter (6) --- Verification of the YAC andACrobes by metaphase FISH --- p.37 / Chapter (7) --- Hybridization efficiency test --- p.38 / Chapter Chapter 3 --- Southern Blot Analysis for Satellite DNA Hypomethylation / Chapter 3.1 --- Introduction --- p.39 -40 / Chapter 3.2 --- Materials and Methods / Chapter (1) --- atients --- p.41 / Chapter (2) --- Mathyl-sensitive restriction enzyme digestion. --- p.42 / Chapter (3) --- Classical satellite 2 DNArobe labeling and hybridization. --- p.42 -43 / Chapter (4) --- Membrane washing and signal detection. --- p.43 / Chapter (5) --- Signal detection and reference ratio determination. --- p.43 -44 / Chapter (6) --- Comparative Genomic Hybridization (CGH) --- p.44 -45 / Chapter 3.3 --- Results / Chapter (1) --- Heterochromatin hypomethylation and 1q12 breakpoint. --- p.45 / Chapter (2) --- Heterochromatin hypomethylation in adjacent hepatitis Infected liver tissue. --- p.46 / Chapter 3.4 --- Discussion --- p.47-51 / Chapter Chapter4 --- ositional Mapping of 1q21 - q22 by Interphase Cytogenetics / Chapter 4.1 --- Introduction --- p.52-53 / Chapter 4.2 --- Materials and Methods / Chapter (1) --- atients --- p.53 / Chapter (2) --- YAC clones --- p.53 -54 / Chapter (3) --- AC clones --- p.55 / Chapter (4) --- Formalin-fixedaraffin-embedded tissue sections pretreatment. --- p.55 / Chapter (5) --- Hybridization --- p.56 / Chapter (6) --- Signal detection --- p.56 -57 / Chapter 4.3 --- Results / Chapter (1) --- Relative copy number gain on YAC examined. --- p.57 -59 / Chapter (2) --- AC findings --- p.60 / Chapter 4.4 --- Discussion --- p.60 -63 / References

by Ray, Kit Ng. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 112-121). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgments --- p.iv / Abbreviations --- p.x / List of Figures --- p.xii / List of Tables --- p.xiv / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Hepatitis B Virus (HBV) --- p.1 / Chapter 1.2 --- Hepatitis B Virus X Protein (HBx) --- p.5 / Chapter 1.2.1 --- The Genomic Structure of HBx --- p.5 / Chapter 1.2.2 --- The HBx Protein Structure --- p.6 / Chapter 1.2.3 --- Subcellular Localization of HBx --- p.7 / Chapter 1.2.4 --- Possible Functions of HBx --- p.8 / Chapter 1.3 --- Etiology of Hepatocellular Carcinoma (HCC) --- p.12 / Chapter 1.4 --- Relationship between HCC and HBx --- p.13 / Chapter 1.5 --- Aims of Study --- p.14 / Chapter 1.6 --- The Basis of Tet-On System --- p.15 / Chapter 1.7 --- The Basis of DNA Microarray --- p.18 / Chapter 1.8 --- The Basis of Two-Dimensional Electrophoresis --- p.20 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Construction of a Tet-On HBx Expressing Cell Model --- p.22 / Chapter 2.1.1 --- Cloning of HBx Gene into pTRE2 Vector --- p.22 / Chapter 2.1.1.1 --- PCR of HBx Gene --- p.22 / Chapter 2.1.1.2 --- Purification of the PCR Product --- p.23 / Chapter 2.1.1.3 --- Restriction Enzyme Digestion --- p.23 / Chapter 2.1.1.4 --- Ligation of HBx into pTRE Vector --- p.24 / Chapter 2.1.1.5 --- Transformation of the Ligation Product into Competent Cells --- p.24 / Chapter 2.1.2 --- Preparation of the Plasmid DNA --- p.24 / Chapter 2.1.2.1 --- DNA Sequencing of the Cloned Plasmid DNA --- p.25 / Chapter 2.1.3 --- Cell Culture of AML12 Cell Line --- p.26 / Chapter 2.1.4 --- Transfection of pTet-On Vector into AML12 Cells --- p.26 / Chapter 2.1.5 --- Selection of the Transfected AML12 Cells by G418 --- p.27 / Chapter 2.1.6 --- Single Clone Isolation --- p.27 / Chapter 2.1.6.1 --- Luciferase Assay for Selection of Highly Inducible Clones --- p.28 / Chapter 2.1.7 --- Second Transfection of pTRE-HBx Plasmid --- p.28 / Chapter 2.1.8 --- Selection of the Transfected Cells by Hygromycin --- p.29 / Chapter 2.1.9 --- Second Single Clone Isolation --- p.29 / Chapter 2.1.10 --- Total RNA Isolation --- p.29 / Chapter 2.1.11 --- DNase I Digestion --- p.30 / Chapter 2.1.12 --- First-Strand cDNA Synthesis --- p.31 / Chapter 2.1.13 --- RT-PCR of HBx Gene --- p.31 / Chapter 2.1.14 --- Northern Blotting --- p.32 / Chapter 2.1.15 --- Preparation of the Probe --- p.33 / Chapter 2.1.16 --- Northern Blot Hybridization --- p.33 / Chapter 2.1.17 --- 3H-Thymidine Incorporation Assay --- p.34 / Chapter 2.1.18 --- Analysis of Cell Cycle by Flow Cytometry --- p.35 / Chapter 2.2 --- Microarray Analysis of Differential Gene Expression upon HBx Induction --- p.35 / Chapter 2.2.1 --- Sample Preparation for Microarray Analysis --- p.35 / Chapter 2.2.2 --- Probe Labelling --- p.36 / Chapter 2.2.3 --- Microarray Hybridization --- p.37 / Chapter 2.2.4 --- RT-PCR of the Candidate Genes --- p.38 / Chapter 2.2.5 --- Northern Blot Analysis of the Candidate Genes --- p.39 / Chapter 2.3 --- Two-Dimensional (2D) Gel Electrophoretic Analysis --- p.40 / Chapter 2.3.1 --- Protein Sample Preparation for 2D Gel Electrophoresis --- p.40 / Chapter 2.3.2 --- First-Dimension Isoelectric Focusing (IEF) --- p.40 / Chapter 2.3.3 --- Second-Dimension SDS-PAGE --- p.41 / Chapter 2.3.4 --- Silver Stain of 2D Gel --- p.42 / Chapter 2.3.5 --- Mass Spectroscopic Analysis --- p.43 / Chapter 2.4 --- Subcellular Localization of HBx --- p.44 / Chapter 2.4.1 --- Cloning of HBx into Green Fluorescent Protein (GFP) Expression Vector --- p.44 / Chapter 2.4.2 --- Transfection of GFP-HBx --- p.44 / Chapter 2.4.3 --- Propidium Iodide (PI) Staining --- p.45 / Chapter 2.4.4 --- Mitochondria Staining --- p.45 / Chapter 2.4.5 --- Subcellular Localization Study using Epi-Fluorescent Microscopy --- p.45 / Chapter 2.5 --- Analysis of Mitochondrial Transmembrane Potential --- p.46 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Construction of Tet-On AML12 Cell Line of HBx Gene --- p.47 / Chapter 3.2 --- Characterization of the HBx-Expressing Cell Model --- p.53 / Chapter 3.2.1 --- 3H-Thymidine Proliferation Assay --- p.53 / Chapter 3.2.2 --- Cell Cycle Analysis --- p.55 / Chapter 3.3 --- Microarray Analysis of Differential Gene Expression Pattern upon HBx Induction --- p.57 / Chapter 3.4 --- Northern Blot Analysis and RT-PCR of the Candidate Genes --- p.65 / Chapter 3.5 --- Differential Protein Expression Pattern under HBx Induction --- p.70 / Chapter 3.6 --- Subcellular Localization of HBx --- p.77 / Chapter 3.7 --- Analysis of Mitochondrial Transmembrane Potential --- p.83 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Conditional HBx-Expressing Cell Model --- p.84 / Chapter 4.2 --- The Effects of HBx in Clone X18 --- p.86 / Chapter 4.2.1 --- Proliferative Effect of HBx --- p.86 / Chapter 4.2.2 --- Deregulation of G2/M Checkpoint by HBx --- p.86 / Chapter 4.3 --- Early Differential Gene Expression due to HBx Induction --- p.88 / Chapter 4.4 --- The Relationship of the Potential Candidate Genes and Cancer Development --- p.90 / Chapter 4.5 --- The Protein Expression Pattern due to HBx Induction --- p.93 / Chapter 4.6 --- The Subcellular Localization of HBx --- p.96 / Chapter 4.7 --- The Possible Involvement of HBx in Mitochondrial Transmembrane Potential --- p.98 / Chapter 4.8 --- Conclusions --- p.101 / Chapter 4.9 --- Future Prospects --- p.104 / Appendix --- p.107 / References --- p.112

Au Chi Chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 157-169). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Table of Contents --- p.ii / Abstract --- p.v / 論文摘要 --- p.vii / Abbreviations --- p.viii / List of Figures --- p.ix / List of Tables --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- General introduction --- p.1 / Chapter 1.2 --- Hepatitis B virus and Hepatocellular carcinoma --- p.3 / Chapter 1.3 --- Pathogenesis of HBV related HCC --- p.6 / Chapter 1.4 --- Current screening test and tumor markers --- p.10 / Chapter 1.5 --- Expressed sequence tag (EST) sequencing --- p.13 / Chapter 1.6 --- Aim of the present study --- p.15 / Chapter 1.7 --- Characterization of KIAA0022 --- p.16 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Construction of liver HCC and normal counterpart libraries --- p.19 / Chapter 2.2 --- Plating out the human liver cDNA libraries --- p.19 / Chapter 2.3 --- PCR amplification of clones human liver cancer and the normal counterpart cDNA libraries --- p.21 / Chapter 2.4 --- Cycle sequencing of cloned human liver cancer and the normal counterpart cDNA libraries --- p.21 / Chapter 2.4.1 --- Dye-primer cycle sequencing (Pharmacia) --- p.21 / Chapter 2.4.1.1 --- Using Pharmacia LBKA.L.F. DNA sequencer --- p.21 / Chapter 2.4.1.2 --- Using Li-Cor 4200 Automated DNA sequencer --- p.22 / Chapter 2.4.2 --- Dye-terminator cycle sequencing (Pharmacia) --- p.22 / Chapter 2.5 --- Sequences analysis --- p.23 / Chapter 2.6 --- Cloning of full-length cDNA of KIAA0022 --- p.24 / Chapter 2.6.1 --- Amplification of KIAA0022 gene using PCR --- p.24 / Chapter 2.6.2 --- Purification of the PCR product --- p.25 / Chapter 2.6.3 --- Ligation --- p.25 / Chapter 2.6.4 --- One Shot® TOP 10 Chemical Transformation --- p.25 / Chapter 2.6.5 --- Small-scale preparation of the plasmid DNA --- p.26 / Chapter 2.6.6 --- Large-scale preparation of the plasmid DNA Table of Contents (continued) --- p.26 / Chapter 2.6.7 --- DNA sequencing of the full-length cDNA of KIAA0022 --- p.28 / Chapter 2.7 --- Northern Hybridization --- p.29 / Chapter 2.7.1 --- The Human multiple tissue Northern Blot --- p.29 / Chapter 2.7.2 --- Synthesis of the radiolabeled DNA probe --- p.29 / Chapter 2.7.3 --- Hybridization of the Northern blot --- p.30 / Chapter 2.8 --- Subcellular localization of KIAA0022 by tagging with green fluorescence protein (GFP) --- p.30 / Chapter 2.8.1 --- Amplification and purification of the KIAA0022 gene product --- p.30 / Chapter 2.8.2 --- Restriction enzymes digestion --- p.31 / Chapter 2.8.3 --- DNA ligation --- p.31 / Chapter 2.8.4 --- Preparation of the Escherichia coli competent cells for transformation --- p.31 / Chapter 2.8.5 --- Transformation of the plasmid DNA into competent Escherichia coli cells --- p.32 / Chapter 2.8.6 --- Small-scale preparation of the plasmid DNA --- p.32 / Chapter 2.8.7 --- Large-scale preparation of the plasmid DNA --- p.32 / Chapter 2.8.8 --- DNA sequencing of the cloned plasmid DNA --- p.33 / Chapter 2.8.9 --- Transfection --- p.33 / Chapter 2.8.10 --- Fluorescence microscopy examination --- p.33 / Chapter 2.9 --- Yeast two-hybrid screening assay --- p.34 / Chapter 2.9.1 --- "Cloning of the KIAA0022 gene into the yeast two-hybrid DNA-BD vector, pGBKT7" --- p.34 / Chapter 2.9.2 --- Small-scale transformation of pGBKT7-KIAA0022 plasmid --- p.34 / Chapter 2.9.2.1 --- Preparation of yeast competent cells --- p.34 / Chapter 2.9.2.2 --- Transformation of the pGBKT7-KIAA 0022 plasmid into the yeast strain PJ69-2A --- p.35 / Chapter 2.9.3 --- Screening a pretransformed library by yeast mating --- p.35 / Chapter 2.9.4 --- β -Galactosidase analysis - colony lift filter assay --- p.36 / Chapter 2.9.5 --- Analysis of yeast plasmid inserts using PCR and DNA sequencing --- p.37 / Chapter 2.9.5.1 --- PCR --- p.37 / Chapter 2.9.5.2 --- DNA sequencing --- p.37 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Results of ESTs sequencing in normal counterpart and HCC libraries --- p.38 / Chapter 3.1.1 --- The sequencing results of the normal counterpart cDNA clones --- p.38 / Chapter 3.1.2 --- Sequencing results of the human liver cancer cDNA clones --- p.41 / Chapter 3.1.3 --- The accuracy of the automated sequencing technique --- p.41 / Chapter 3.1.4 --- Catalogue of normal counterpart ESTs --- p.45 / Chapter 3.1.5 --- Catalogue of liver cancer ESTs --- p.47 / Chapter 3.2 --- Identification of genes differentially expressed in HCC using in silico method --- p.115 / Chapter 3.3 --- Sequence analysis of KIAA0022 --- p.121 / Chapter 3.3.1 --- Structural analysis of KIAA0022 --- p.121 / Chapter 3.3.2 --- Homology alignment --- p.122 / Chapter 3.4 --- Tissue distribution and expression profile of KIAA0022 using Northern blot analysis --- p.132 / Chapter 3.5 --- Subcellular localization of the KIAA0022 tagging by green fluorescence protein --- p.134 / Chapter 3.6 --- Yeast two-hybrid screening assay --- p.136 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Large-scale partial cDNA sequencing --- p.138 / Chapter 4.2 --- Characterization of ESTs --- p.139 / Chapter 4.3 --- Identification of genes differentially expressed in liver cancer using Poisson probability --- p.143 / Chapter 4.4 --- Characterization of KIAA0022 --- p.154 / Reference --- p.157 / Appendix --- p.170

Tang Wan-yee. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 192-212). / 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.

by Sung Ying-Man, Mandy. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 112-125). / Abstracts in English and Chinese. / Acknowledgments --- p.I / List of Abbreviations --- p.II / Abstract --- p.IV / 摘要 --- p.VII / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Epidemiology --- p.1 / Chapter 1.2 --- Aetiological factors --- p.4 / Chapter 1.2.1 --- Hepatitis B infection --- p.4 / Chapter 1.2.2 --- Aflatoxin exposure --- p.5 / Chapter 1.2.3 --- Alcohol consumption --- p.5 / Chapter 1.2.4 --- Genetic risk factors --- p.6 / Chapter 1.3 --- Aims of the study --- p.7 / Chapter Chapter 2 --- Polymorphisms of candidate genes in Interleukin- signalling pathway in HCC --- p.6 / Chapter 2.1 --- Introduction --- p.9 / Chapter 2.2 --- Materials and Methods --- p.15 / Samples and Genomic DNA isolation --- p.15 / PCR-PFLP --- p.15 / dHPLC --- p.16 / Direct Sequencing --- p.17 / Stattistical Analysis --- p.17 / Chapter 2.3 --- Results --- p.25 / Chapter 2.3.1 --- Known IL6 polymorphisms --- p.25 / Chapter 2.3.2 --- IL-6R and gp130 polymorphisms --- p.28 / Chapter 2.3.3 --- Stat-3 polymorphisms --- p.29 / Chapter 2.3.4 --- SOCS-1 polymophisms --- p.32 / Chapter 2.3.5 --- Mutation screening of IL-6 gene --- p.34 / Chapter 2.3.6 --- Mutation screening of SOCS-1 gene --- p.39 / Chapter 2.4 --- Discussion --- p.40 / Chapter 2.4.1 --- Interleukin-6 --- p.40 / Chapter 2.4.2 --- Gp130 and IL6-R --- p.43 / Chapter 2.4.3 --- STAT-3 --- p.44 / Chapter 2.4.4 --- SOCS-l --- p.46 / Chapter Chapter 3 --- Methylation status of SOCS-1 gene in Chinese HCC patients / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Methods and Materials --- p.53 / Tissue Sampling --- p.53 / Methylation specific PCR (MSP) --- p.53 / Chapter 3.3 --- Results --- p.55 / Chapter 3.4 --- Discussion --- p.56 / Chapter Chapter 4 --- Polymorphisms of enzyme encoding genesin steroidogenesis in Chinese / Chapter 4.1 --- Introduction --- p.59 / Chapter 4.1.1 --- Steroid 5a reductases (SRD5A) --- p.62 / Chapter 4.1.1a --- Steroid 5a reductase type II (SRD5A2) --- p.63 / Chapter 4.1.1b --- Steroid 5a reductase type I (SRD5A1) --- p.65 / Chapter 4.1.2 --- Cytochrome P450al7 (CYP17) --- p.67 / Chapter 4.1.3 --- "Cytochrome P450, family 1，subfamily A polypeptide1 (CYP1A1)" --- p.69 / Chapter 4.1.4 --- "Cytochrome P450, subfamily IIIA (niphedipine oxidase) polypeptide 4 (CYP3A4)" --- p.71 / Chapter 4.2 --- Materials and Methods --- p.74 / Samples and Genomic DNA isolation --- p.74 / PCR-PFLP --- p.74 / Direct Sequencing --- p.74 / Statistical Analysis --- p.74 / Chapter 4.3 --- Results --- p.77 / Chapter 4.3.1 --- SRD5A2 --- p.77 / Chapter 4.3.2 --- Linkage Disequilibrium in SRD5A2 gene --- p.83 / Chapter 4.3.2 --- SRD5A1 --- p.84 / Chapter 4.3.3 --- CYP17 --- p.87 / Chapter 4.3.4 --- CYP1A1 --- p.89 / Chapter 4.3.5 --- CYP3A4 --- p.92 / Chapter 4.3.6 --- Logistic regression --- p.95 / Chapter 4.4 --- Discussion --- p.96 / Chapter 4.4.1 --- SRD5A2 --- p.96 / Chapter 4.4.2 --- SRD5A1 --- p.99 / Chapter 4.4.3 --- CYP17 --- p.101 / Chapter 4.4.4 --- CYP1A1 --- p.103 / Chapter 4.4.5 --- CYP3A4 --- p.106 / Chapter 4.4.6 --- Logistic Regression --- p.107 / Chapter Chapter 5 --- Conclusions and Future Prospect --- p.108 / Chapter 5.1 --- Conclusions --- p.108 / Chapter 5.2 --- Future works and prospect --- p.111 / References --- p.113