The Relative Study Of Protein Z And Genetic Variations In The Gene Protein Z Gene In Patients With Acute Ischemic Stoke

Background：With the increase of older people, the incidence of cardiac-cerebral thrombosis diseaseis increasing, and cardiac-cerebral thrombosis disease has become one of the most commoncauses of death. Thrombosis is a complex pathological process, including the mechanism ofvascular factors, blood coagulation, anticoagulation and fibrinolysis disorders. Investigating the pathogenesis of thrombosis, it may have an important guiding significance in theclinical diagnosis and treatment of thrombosis. Protein Z (protein Z, PZ) is a vitaminK-dependent blood coagulation factors, protein Z-dependent protease inhibitor is (ProteinZ-dependent protease inhibitor, ZPI) inhibit the coagulation factor Xa cofactor, PZ inhibitthe coagulation factor can be improved ZPI Xa activity, play a role in regulating blood andbi-thrombotic diseases. Recently a small amount of literature at home and abroad has beenreported that the plasma PZ levels and gene polymorphism is related to cardiac-cerebralthrombosis disease, but results vary. This is the first in the Chinese han population toexplore the correlation of plasma PZ level and gene polymorphism with cardiac-cerebralthrombosis disease and its possible mechanism, for providing a theoretical basis ofprediction, diagnosis and treatment of linical cardiac-cerebral thrombosis disease.Objectives：1. To explore the correlations of plasma PZ levels and acute ischemic stoke. 2. To explore the relationships of PZ gene polymorphism and acute ischemic stoke.3. To reveal the possible relationship between plasma PZ levels and its intron F G79Aand Promoter A-13G gene polymorphism.4. To fill the blank of study on PZ with acute ischemic stoke disease genepolymorphism in Chinese population.Methods：The first part Testing the plasma PZ level in patients with acute ischemic stoke.The blood samples of acute ischemic stoke patients and healthy controls wereCollected.The plasma level of PZ was detected by using ELISA assay. The experiment wasdivided into2groups group：acute ischemic stoke (Sub-group and recurrent group), normalcontrol group. In strict accordance with the PZ ELISA kit instructions to complete theexperiment process. Accomplished statistical analysis and explored the change of plasmaPZ level in patients with acute ischemic stoke.The second part Detection PZ promoter A-13G and two in G79A polymorphism inintron F.The experiment was divided into2groups: acute ischemic stoke group, normal controlgroup. Each blood sample was completed collection, The blood samples in patients withacute ischemic stoke and healthy control group were collected. The genomic DNA wasextracted from white cells in each group of blood samples. To amplify the genomic DNAby Polymerase Chain Reaction (PCR), and identify PZ gene polymorphism by genesequencing, detection PZ promoter A-13G and two in G79A polymorphism in intron F.The third Part Analysis of PZ plasma levels and its correlation in PZ genepolymorphism.PZ plasma levels were compared using t test statistics; polymorphism genotype and allele frequencies were compared using chi-square test; calculated for each group of alleleand genotype frequencies using the Hardy-Weinberg equilibrium test; PZ plasma levels ofits genes polymorphism using the Spearman rank correlation coefficient indicates; PZgenotype and allele with AIS risk prediction using odds ratios (OR) and95%confidenceintervals (CI) to represent.Results：The first part The change of plasma PZ level in patients with acute ischemic stoke1. The plasma PZ level was significantly lower (P<0.01) in patients with acute ischemicstoke than that in healthy controls.2. AIS recurrence group PZ levels was lower the initial group, a significant statisticaldifference (P<0.01).The second part Distribute PZ promoter A-13G and two in G79A polymorphism inintron F.1. The AA genotype and A allele frequency of PZ in intron F G79A was significantlylower (P <0.05) in patients with acute ischemic stoke than in healthy controls.2. G allele frequency of PZ in intron F G79A was significantly higher (P<0.05) inpatients with acute ischemic stoke than in healthy controls.(P<0.05).3. There was statistically significant (AG, GG, AA) between genotype groups G79AAIS sites with the control group(P=0.05).4. AIS group A-13G locus allele frequencies and genotype (AG, GG, AA) notsignificantly different from the control group (P>0.05).5. Control group PZ promoter A-13G G allele frequency was significantly higher than theEuropean Caucasian population (60.7%vs13.1%), PZ intron F G79A in the A allelewere significantly higher than the European Caucasian population (61.1%vs40.7%).The third Part PZ plasma levels correlated with its gene polymorphism.1. The PZ level in AA genotype of the PZ in intron F G79A was significantly decreased(P<0.05). 2. The PZ level in GG genotype of the PZ in intron F G79A compared with normalcontrols was higher (P<0.05).3. AIS group PZ start no significant difference in the level of sub-A13G genotype andthe control group PZ (P>0.05).4. The acute ischemic stoke was significantly(P<0.01)related to these risk factors ofsmoking, hypertension, hyperlipidemia and diabetes.Conclusions：1. The plasma PZ levels were correlated with acute ischemic stoke, it tips PZ deficiencymay be a risk factor for acute ischemic stroke disease exists.2. PZ promoter A-13G locus A allele frequencies and genotype had no correlation withacute ischemic stoke, reflecting PZ gene A-13G polymorphism may not be involved ofdevelopment in the disease.3. The AA genotype and A allele frequency of PZ gene G79A was negatively correlatedwith acute ischemic stoke, reflecting the AA genotype as a protective risk factors for acuteischemic stoke.4. The plasma PZ levels of the AA genotype among PZ gene G79A locus AA, GG andGA genotypes were negative correlation with acute ischemic stoke, indicating that the levelPZ of the AA genotypes may reduce the risk of thrombotic disease by way ofdownregulating expression in thrombotic diseases, revealing AA genotype may reflect aprotective risk factors in patients with AIS.5. PZ gene mutation racial differences exist, the Chinese Han population PRD mutationrate was significantly higher than the European Caucasian population.6. The acute ischemic stoke was significantly related to these risk factors of smoking,Changing these risk factors can reduce the incidence and recurrence rates AIS.