| BackgroundCoronary heart disease (CHD) is associated closely with the coagulation system. The disorders of blood coagulation are often found in CHD patients. The rupture of atheromatous plaque and thrombosis is the major cause of acute myocardial infarction and unstable angina. Coagulation factors and antifibrinolytic mass are important in atherosclerosis.Coagulation factor XIII (FXIII) is also called fibrin-stabilizing factor, its A-subunit is a transglutaminase. Plasma FXIII is a tetramer (A2B2), whereas the cellular FXIII is a homodimer of two A subunits. FXIII is activated by thrombin, which cleaves the A-subunit between Arg37 and Gly38. Activated FXIII catalyses the covalent cross-linking of the y and a chains of fibrin, increasing the mechanical strength of fibrin and its resistance to fibrinolysis.Recently a G→T point mutation in codon 34 of exon 2 which codes for aValine→Leucine(FXIII Val34Leu)change in the A-subunit was reported to protect against venous thrombosis but predispose to hemorrhage. Studies have demonstrated that fiber mass/length ratio and porosity were reduced in fibrin clot formed by FXIII 34Leu subjects which led to ineffective fibrin cross-linking or alternatively deficient cross-linking of other natural substrates for FXIII.The present studies for FXIII Val34Leu are almost around the white people. The association of FXIII Val34Leu with CHD and myocardial infarction is uncertain especially in Chinese. We investigated the FXIII Val34Leu using polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) technique. Our objective is to study the distribution of FXIII Val34Leu in Chinese people and its relationship with CHD and myocardial infarction.Material and Methods:1. Subjects:398 people of the Han nationality without blood relationship were recruited to the study. Among them, 195 CHD patients (154 male and 41 female, the average age was 64.25 ± 9.43) from the Cardiovascular Department of the Second Affiliated Hospital, Zhejiang University College of Medicine. All patients were confirmed by angiography: at least 50% obstruction of one major coronary artery. The CHD group was divided into myocardial infarction(MI) group(87) and none-myocardial infarction group(108) using criteria of by the World Health Organization (WHO) ;203 healthy control subjects(145 male and 58 female, the average age was 62.54±8.77) were from patients with normal angiographies and general practice registers, having no manifestation of CHD by history, physical examination and electrocardiogram (ECG).2. Methods:2.1 Collection of clinical features: All subjects were enquired in detail for history ofhypertension, diabetes, hyperlipemia, smoking. The stature, weight and major biochemical data were also determinated.2.2 FXIII Val34Leu genotype was determined by PCR-SSCP: (1) The primers of Kohler and Kangsadalampai were referenced, the forward primer: 5'-GAC CTT GTA AAG TCA AAA ATG TC-3'and the reverse primer: 5'-TAC CTT GCA GGT TGA CGC CCC GGG GCA CTA-3'. (2) The DMA template was obtained from 2-5ml venous blood using phenol/chloroform separation. (3) PCR was carried out using a 25ul reaction mix containing 10pmol of each primer ,100ng of genomic DNA,200uM of each dNTP,2.5ul of 10×buffer, 1-3uM of MgCI2,2U of Taq DMA polymerase. The PCR cycles were modified as follows:5 min initial at 94 ℃,followed by 30 cycles of 30 sec of denaturation at 94℃,30 sec of annealing at 53℃ and 30 sec of extension at 72℃ followed by a final 5 min extension step at 72℃.(4) SSCP: PCR products were denatured and loaded onto a 12% polyacrylamide gel (29:1) containing 5% glycerol for electrophoresis. Gels were visualized by ethidium bromide (EB) staining and analyzed with the Gel Imaging System. 3. Statistical analysis:Using the SPSS10.0 for windows statistical package. Count data were performed by Student's t-test and measure data were performed by chi-squared test. Genotype and allele frequencies between groups and the agreement with Hardy-Weinberg equilibriu...
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