| Background and Purpose:It is known to all that ischemic cerebravascular diseases have high attack rate and mortality,which will result in heavy burden to family and society.Studies on ICVD risk factors are the important researching directions,especially genetics aspects, which can identifit ICVD predisposing genes.So that identify high risk group of ICVD and provide more energetic preventions and cure strategies for ICVD patients.Clotting and fibrinolytic dysfunction is one of the pathogenesis of ICVD.As the major inhibitor of fibrinolytic system,the plasma level of plasminogen activator inhibitor-1 reflects fibrinolytic function of organisms.Studies indicate that which is seriously participate in thrombus information during ischemie cerebralvascular disease course.If the coding gene mutated,which will caused the corresponding plasma level set up or down,thereby fibrinolysis dysfuncted.At present,more polymorphic sites of plasminogen activator inhibitor-1 have been reported,especially 675 4G/5G polymorphism site,as well is related to plasma level of plasminogen activator inhibitor-1.PAI-1 antigen activity is siginificantly impacted by nature and nurture factors, researchers still keep controversial about whether PAI-1 polymorphisms is the independent risk factor of ICVD,lack of large sample experiment,either.To further explore the relation between PAI-1 polymorphism,the genetic risk factor of cerebrovascular,and ICVD.The study will explore PAI-1 675 4G/5G,-844G/A gene polymorphisms in Henan han populations,to investigate the distribution of PAI-1 gene 675 4G/5G and -844G/A polymorphisms in Henan han population,so that to explore the role PAI-1 polymorphisms play in the development process of ICVD. Aim to further approach the etiopathogenisis and pathogenesis of ICVD,to provide new strategy for prevention ICVD accordingly.Methods:408 cases as patient group with ischemic cerebrovascular disease were enrolled from December 2006 to July 2008 in departments of neurology in the hospitals which be above second order of grade A in Henan province(220 men and 188 women,mean average 59.4±11.6 years).Diagnosis of ICVD reference to the diagnostic criteria on stroke which be revised by WHO in 2006.418 unrelated controls were selected from old subjects in hospital simultaneously(226 men and 192 women,mean age 57.8±11.1 years old).Physical and laboratory examination,history taking to be carried out in the group,meanwhile exclude the people who have the history of cardio -cerebrovascular disease.The study population were people in Henan han.The statistical analysis showed that the difference about distribution of age and sex between the two groups was not significant by statistic analysis.The people who had epilepsy,tumor,liver or kidney dysfunction,serious malnutrition were excluded from the two groups.Peripheral venous blood samples were collected from every object.Genomic DNA was extracted from white cells adopting standard Phenol-Chloroform method. Design the primers,and plasminogen activator inhibitor-1 675 4G/5G site,we performed allele-specific polymerase chain reaction(ASP) amplification and performed polymerase chain reaction-restrition fragement length polymorphism (RFLP) to detect the PAI-1-844G/A site polymorphism,the final products were electrophoresed on 2.0%agarose gels,then detect the results and take pictures in the ultraviolet imaging instrument.All the experimental data were input and establish database in SPSS 13.0 software,the frequencies of genotype and allele were estimated by the gene-counting method and compared using the x~2 analysis.The x~2 test were confirmed to sure these genotypes were compatible with genetic equilibrium and check the group comparison of genotypic and allele frequency,andα=0.05 was used as a statistical test standards.Results:1.In Henan han population,the genotype distribution of PAI-1 gene 675 4G/5G and -844G/A polymorphisms were compatible with the Hardy-Weinberg equilibrium.2.Among the subjects of the Henan han population,the mutation frequencies of PAI-1 gene 675 4G/5G,-844G/A site might vary with different ethnic groups or geographic regions.3.In Henan han population,the distribution of allele and genotype in 675 4G/5G and -844G/A polymorphism of PAI-1 had no significant difference between the cases of control group and ICVD patients.Conclusion:1.There are the PAI-1 gene 675 4G/5G and -844G/A polymorphisms in Henan han population,and the frequencies of the PAI-1 gene 675 4G/5G,-844G/A site mutations might vary with different ethnic group or geographic regions.2.PAI-1 gene 675 4G/5G and -844G/A polymorphisms may not be ICVD genetic risk factors in Henan han population. |
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