BackgroundCerebral infarction(CI)is a common cerebrovascular disease with high morbidity and mortality,It has become a great danger to human health .CI may arise from interactions between environmental and genetic factors in general.The imbalance of coagulation and fibrinolysis is an important cause participating in the pathogenesis of CHD and CI.Factorâ…¦(Fâ…¦) is the first enzyme in the extrinsic pathway of the blood coagulation system.Activation of the extrinsic coagulation pathway plays a key role in hemostasis,and Fâ…¦participates the intrinsic coagulation pathway.Some studies have found the Fâ…¦in plasma should be associated with CHD and ICVD.In 1991,Green reported Fâ…¦gene polymorphisms of R353Q could influence plasma Fâ…¦coagulant activity.Carrier of the Q allele(encoding GLn353)have levels of Fâ…¦c lower than than those of R allele (encoding Arg 353) and have reduced acute thrombotic events.Fâ…¦Q allele is a protective factor of the arterial thrombotic diseases.but the occurrence of Fâ…¦Q allele is rare in general population in studies abroad.the viewpoint that the Q is a protective factor in thrombotic diseases seems uncertain .The researcher about association of Fâ…¦R353Q with CI is scarce. We adopted PCR-RFLP to study whether the Fâ…¦R353Q polymorphism is associated with risk of CI in the Han population.Study population1.The case group include100 individuals with CI chosen from the patients who were in neurological department, the second Hospital of Jilin University during September 2007 to December 2007. Clinical diagnosed as CI by CT or MRI and conformed with the 1995 edited Standard of the 4th Meeting to cerebrovascular disease.69 male, 31 female aged extent: 33-83 years old. All of them are Han population.2.Controls 106 unrelated heathy controls were selected from subjects in outpatient who underwent regular check-up examinarion. 54 male, 52 female aged extent: 30-80 years old. The cases of Control group had no history of CHD,EH and DM.All of them are Han population.MethodsDNA was extracted from venous white blood cells. PCR amplifies the exon8 region of Fâ…¦gene , fragments are 312bp. Sense primer :5′-GGG AGA CTC CCC AAA TAT CAC-3'Antisense primer:5'-ACG CAG CCT TGG CTT TCT CTC-3'.The action system of PCR is 50μl , including DNA template 10μl, primer (20um)1μl, TaKaRa Taq(5U/ul) 0.5ul,10×PCR Buffer(Mg2+Plus) 5ul,dNTP Mixture (2.5mM) 4ul. PCR reaction condition: 94℃/5min , force-degeneration;94℃/55s,55℃/45s,72℃/ 1min,35circles;72℃program- dation 5 min.PCR production was identified by 2% agarose gel electro- phoresis and ultraviolet transilluminator. Genotypic identification in the exon8 region of Fâ…¦gene : After PCR products were purified by the purification kit, they were digested by the restriction enzyme MSPâ… . The action system is 20ul: PCR product 10ul, MSPI 1ul,10×T Buffer 2ul,0.1%BSA 2ul, distilled water 5ul.37℃incubation overnight. The result was identified by 2% agarose gel electrophoresis and ultraviolet transilluminator and wrote genotypes.Statistical analysis: Genotype and allele frequencies were estimated by the gene-counting method and compared using Hardy-weinberg equilibrium was confirmed with the X2 test.Genotype and allele frequencies were estimated by compared using the X2 analysis. All statistical analyscs were performed with SPSS 11.0 software.α=0.05.ResultsThere is Fâ…¦R353Q polymorphism in the Han population.R,Q allele and genotype of RR, RQ genotype can be founded in the control group and the CI group .The fragment length of RR genotype separately was 205bp,67bp and 40bp , the fragment length of RQ genotype was 272bp,205bp,67bp and 40bp;R allele genotype frequencies are separately 94.33% and 95.5% in the control group and the CI group ,Q allele genotype frequencies are separately 5.67% and 4.5% in the control group and the CI group . By Statistical analysis, the result is there were no significant differences in the R353Q polymorphism mutation between the cases of control group and CI patients(P>0.05 ).Q allele genotype is not relativity with the CI. It can be seen that the relationship between polymorphism and thrombus disease is still discussing. the frequ- encies of these two gene mutation might vary with different ethnic group or geographic regions.Conclusions1.There are the Fâ…¦R353Q polymorphism in Han population. RR, RQ genotype and R,Q allele can be founded in the control group and the cerebral infarction group .2. Unsuppor the viewpoint that the Q allele is a protective factor in CI .3.Frequencies of these two gene mutation might vary with different ethnic group or country.4.It can be seen that the relationship between Fâ…¦R353Q polymorphism and cerebral infarction is still discussing. We are going on deepening to research synergistic action about Fâ…¦R353Q polymorphism and other risk factor with cerebral infarction。...
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