| Objective To test the expression of polymorphism of promoter gene-1208I/D of TF in patients with venous thromboembolism(VTE) and analyze the correlation between the polymorphism of the gene and VTE.To test the antigen and the activity of plasma TF and discuss the relationship between the levels of TF and polymorphism of the gene.MethodsFirstly,the collection of samples.Patients,both outpatients and inpatients,are from the departments of vascular breast,respiratory and emergency in the Second Hospital affiliated with Shanxi Medical University from March 2006 to July 2007.Among these patients,there are 96 cases of DVT with 54 males and 42 females and their average age is 42.81±16.45;there are 14 cases of PTE with 6 males and 8 females and their average age is 51.44±18.24.Samples of the control group are the healthy medical examinees in the medical examination center of our hospital healthy.There are 59 cases in total with 31 males and 28 females and their average age is 45.2±17.31.Secondly,the extraction of genomic DNA.Genomic DNA in the single nuclear cell was extracted from all samples with Phenol-chloroform.Thirdly,amplification of PCR and analysis of the product.Polymorphism of TF promoter -1208I/D gene was tested by the means of polymerase chain reaction(PCR) and agarose gel electrophoresis.Genotypes of objects of the study were distinguished according to strip electrophoresis and the frequencies of the genotype were counted.Fourth,gene sequencing.Gene of some D/D and I/I homozygous samples were sequenced to verify the existence of two alleles.Fifth,plasma TF antigen and activity of 55 cases of DVT and 28 cases of control group were measured separately with the methods of ELISA and chromogenic substrate,analyzing the correlation of antigen and activity among the different genotypes.Sixth,a software of SPSS13.0 was used for the statistical analysis and gene counting method for calculating genotype frequency and allele frequency of each group.(?)+s was used to indicate the measurement data,t test to comparing the two groups of means and x~2 test to test the count data.Antigen and activity of the three genotypes in each group were compared with the analysis of variance and there was statistical significance when p <0.05.Results1.The mutation of the promoter gene -1208I/D of TF is insertion or deletion of 18bp base (-AGCTAAACGAGATATGTA-),with three genotypes D/D,I/D and I/I.Genotypes include D/ D-type 67.8%,I/D-type 25.4%and I/I-type 6.8%in the control group,D/D62.5%,I/D 29.8%, and I/I 8.3%in DVT group,D/D57.1%,I/D 35.7%and I/I 7.1%in PTE group.D allele frequency and the I allele frequency were 80.5%and 19.5%respectively in the control group, 77.1%and 22.9%in DVT group and 75.0%,25.0%in PTE group with p > 0.05.2.TF antigen in VET and control groups were 22.745±11.411(pg/mL) and 24.012±11.697 (pg/mL) respectively,p=0.637;TF activity were 4.469±4.630(pM) and 1.633±0.648(pM) respectively,p<0.001.3.TF antigen levels of three genotypes D/D,I/D and I/I were 21.437±9.814(pg/mL), 20.816±10.998(pg/mL) and 38.063±3.182(pg/mL) respectively in VET group,21.289±9.856 (pg/mL),23.178±7.125(pg/mL) and 41.667±18.950(pg/mL) in the control group.VET Group D/D,I/D and I/I genotype TF activity of the three F=2.129,p=0.129;in the control group F=0.212,p=0.811.Conclusions1.There is no significant correlation between gene polymorphism of promotor -1208I/D of TF and VET,and the gene may not be a susceptibility gene of VET.2.The increase of TF activity is one of risk factors inducing VET.3.There is an increase in plasma TF antigen of I/I homozygous genotype carriers.Gene polymorphism of promoter-1208I/D of TF can affect the gene expression of TF.4.It would be necessary to further study the gene polymorphism of TF.
|
PDF Full Text Request