Yung Chung Ming Edmund. / Thesis submitted in: September 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 44-54). / Abstracts in English and Chinese. / Title Page / Contents / Abstract / 摘要 / List of Figures and tables / Abbreviations / Text / References / Chapter Chapter 1: --- General Introduction / Chapter 1.1 --- An overview to the study of CTLA-4 gene in childhood Graves' disease (GD) / Chapter 1.1.1 --- "Graves' disease 226}0ؤ features, incidence, aetiology and pathogenesis page" / Chapter 1.1.1.1 --- GD features 226}0ؤ from clinical to laboratory --- p.1 / Chapter 1.1.1.2 --- GD incidence - from adult to children --- p.2 / Chapter 1.1.1.3 --- GD aetiology - from environment to genes --- p.3 / Chapter 1.1.1.4 --- GD pathogenesis - from auto-antigen to autoantibody --- p.4 / Chapter 1.1.2 --- CTLA-4 gene study in Graves' disease --- p.5 / Chapter 1.1.3 --- Conclusion --- p.6 / Chapter 1.2 --- "Objectives, hypothesis and planning of the study" --- p.7 / Chapter 1.2.1 --- Objectives --- p.7 / Chapter 1.2.2 --- Hypothesis --- p.7 / Chapter 1.2.3 --- Planning --- p.7 / Chapter Chapter 2: --- Literature Review --- p.8 / Chapter 2.1 --- The CD28 / CTLA-4: B7 co-stimulatory pathway and Graves' disease --- p.8 / Chapter 2.1.1 --- Overview of co-stimulation and T cell activation --- p.8 / Chapter 2.1.2 --- Overview of the CD28 gene --- p.9 / Chapter 2.1.3 --- Overview of the CTLA-4 gene --- p.10 / Chapter 2.1.4 --- Co-stimulation and Graves' disease --- p.13 / Chapter 2.2 --- The study of CTLA-4 gene polymorphism in Graves' disease --- p.14 / Chapter Chapter 3: --- Methodology --- p.16 / Chapter 3.1 --- Recruitment of subjects --- p.16 / Chapter 3.1.1 --- Recruitment of cases --- p.16 / Chapter 3.1.2 --- Recruitment of controls --- p.16 / Chapter 3.1.3 --- Ethical approval --- p.17 / Chapter 3.2 --- Peripheral blood collection and genomic DNA preparation --- p.17 / Chapter 3.2.1 --- Peripheral blood collection --- p.17 / Chapter 3.2.2 --- White blood cell harvesting --- p.17 / Chapter 3.2.3 --- White blood cell digestion --- p.17 / Chapter 3.2.4 --- DNA extraction --- p.17 / Chapter 3.3 --- Polymerase Chain Reaction (PCR) amplification of CTLA-4 gene exon one --- p.18 / Chapter 3 .4 --- PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis of CTLA-4 gene codon 17 A/G dimorphism --- p.19 / Chapter 3.5 --- PCR-Single Strand Conformational Polymorphism (PCR-SSCP) analysis of of CTLA-4 gene codon 17 A/G dimorphism --- p.21 / Chapter 3.5.1 --- Preparation of SSCP gel and buffer --- p.21 / Chapter 3.5.2 --- ´5ةend labelling of forward PCR primer --- p.21 / Chapter 3.5.3 --- Preparation of PCR fragment for SSCP analysis --- p.21 / Chapter 3.5.4 --- SSCP analysis --- p.22 / Chapter 3.5.5 --- Autoradiography --- p.22 / Chapter 3.6 --- Sequence confirmation of the SSCP fragment by PCR cycle sequencing --- p.22 / Chapter 3.6.1 --- Preparation of sequencing template from SSCP fragment --- p.22 / Chapter 3.6.2 --- PCR cycle sequencing --- p.23 / Chapter 3.6.3 --- Preparation of cycle sequencing products for electrophoresis --- p.23 / Chapter 3.6.4 --- Sequencing by capillary electrophoresis (CE) --- p.24 / Chapter 3.7 --- Statistical analysis --- p.24 / Chapter Chapter 4: --- Results and Data Analysis --- p.26 / Chapter 4.1 --- Results --- p.26 / Chapter 4.1.1 --- Demographic data of case and control subjects --- p.26 / Chapter 4.1.2 --- PCR amplification of CTLA-4 gene exon one --- p.26 / Chapter 4.1.3 --- PCR-RFLP analysis of CTLA-4 gene codon 17 A/G dimorphism locus --- p.26 / Chapter 4.1.4 --- PCR-SSCP analysis of CTLA-4 gene codon 17 A/G dimorphism locus --- p.29 / Chapter 4.1.5 --- PCR cycle sequencing of the SSCP fragments --- p.31 / Chapter 4.2 --- Data analysis --- p.32 / Chapter 4.2.1 --- Overview of data --- p.32 / Chapter 4.2.2 --- CTLA-4 exon one polymorphism analysed with respect to sex --- p.32 / Chapter 4.2.3 --- CTLA-4 exon one polymorphism in patients with Graves' disease and controls --- p.34 / Chapter Chapter 5: --- Discussion --- p.36 / Chapter Chapter 6: --- Summary and Conclusions --- p.42