| Objective: To detect the distribution of the SNP polymorphic site rs4950928 of CHI3L1 gene,and the SNP site rs1671153,rs1654419 of GP6 gene in the susceptible population of coronary artery disease by PCR technique and DNA direct sequencing method,comparing the SNP loci alleles distribution frequency in coronary artery disease group and normal control group,and results revealed the relationship between the SNP polymorphism site of CHI3L1,GP6 gene and coronary artery disease.Methods: 1560 patients with coronary artery disease were recruited from Hebei Cangzhong Pepole Hospital from January 2013 to December 2015 years by coronary angiography,coronary CT and color doppler ultrasound diagnosis.Meanwhile,900 subjects were collected as control group with matching gender and age.All the patients were excluded diseases such as infection,tumor,diabetes,severe congenital artery disease,kidney disease,autoimmune diseases,myocarditis,severe trauma.,etc.Take 5 ml sodium citrate anticoagulant blood to determine the fibrinogen and D dimer.Take 5 ml coagulated blood used for the determination of serum index and inflammation factors.Take 5ml EDTA-K2 anticoagulant blood to extracte Genomic DNA of each subject.Primers were designed with biological software Primer7.0 and compared with DNA sequence in the GeneBank.PCR technology was applied to amplify target gene fragments and direct sequencing method was used to analyze the DNA polymorphism of CHI3L1 and GP6 gene.All the analyses were performed with the SPSS 19.0 statistical analysis package.The statistical indexes were all showed by mean ± standard deviation(mean ± SD).The frequency of genotype and allele were calculated with gene counting method.The differences of the frequencies of the alleles and the genotype Between CHD group and control group were analyzed by constructing Chi-square(χ2)analysis.The relationship of CHI3L1 and GP6 genotype SNP site polymorphisms and gastric cancer was analyzed by stepwise Logistic regression analysis.P <0.05 for the difference was significant.Results:1 Compared with the control group,The gender,age and drinking history of Coronary artery disease group had no statistical difference(P > 0.05),while the smoking history and family history of coronary artery disease group was obviously higher than those of normal control group,,the difference has significant statistical(P < 0.05).2 The serum concentrations of TG,LDL,Glu,Fbg,D-D,HCY,HsCRP,CHI3L1,TNF-α,IL-1,GP6 and VCAM were significantly higher in Coronary artery disease group than those of control group.There were significant differences(P<0.05).The serum concentration of HDL was significantly lower than the normal control group,the difference was statistically significant(P < 0.05).3 The frequencies distribution of CC,CT,TT genotypes in SNP site rs4950928 of CHI3L1 gene in coronary artery disease group were 0.7808,0.1558 and 0.0634 respectively,the frequency of CT+TT genotypes was0.2192,the frequency of T allele was 0.1413,while the frequencies distribution of CC,CT,TT genotypes in control group were 0.9078,0.0756 and 0.0166 respectively,the frequency of CT+TT genotypes was 0.0922,the frequency of T allele was 0.0544.The frequencies of TT,CT genotype,CT+ TT gene type and T allele were obviously higher than those of normal control group,the differences was statistically significant between the two groups(X2 =38.38,33.14,64.42,88.37,P<0.05).4 The frequencies distribution of CC,CA,AA genotypes in SNP site rs1671153 of GP6 gene in coronary artery disease group were 0.9276,0.0500 and 0.0224 respectively,the frequency of CA+AA genotypes was 0.0724,the frequency of A allele was 0.047,while the frequencies distribution of CC,CA,AA genotypes in control group were 0.9466,0.0467 and 0.0067 respectively,the frequency of CA+AA genotypes was 0.0533,the frequencyof A allele was 0.0306.The frequencies of AA genotype,AA gene type and A allele were obviously higher than those of normal control group,the differences was statistically significant between the two groups(X2=8.723,8.811,P<0.05).5 The frequencies distribution of TT,TC,CC genotypes in SNP site rs1654419 of GP6 gene in coronary artery disease group were 0.9288,0.0411 and 0.0301 respectively,the frequency of TC+CC genotypes was 0.0712,the frequency of C allele was 0.0506,while the frequencies distribution of TT,TC,CC genotypes in control group were 0.9478,0.0378 and 0.0144 respectively,the frequency of TC+CC genotypes was 0.0522,the frequency of C allele was 0.0333.The frequencies of CC genotype,TC+CC gene type and C allele were obviously higher than those of normal control group,the differences was statistically significant between the two groups(X2=5.96,8.05,P<0.05).6 Stepwise Logistic regression:(1)Taken coronary artery disease as dependent variable(yes=1,no=0)and genotype(CC=0,CT=1,TT=2),TNF-α,IL-1,Hs-CRP,family history(yes=1,no=0),WC,BMI,HCY,Fbg,D-Dimer,TC,TG,HDL,LDL as independent variables to make Stepwise regression analysis,found that the genotype,LDL is closely related to the onset of coronary artery disease,the genotype and LDL can significantly increase the risk of coronary artery disease(CHD),they are independent risk factors for coronary artery disease pathogenesis,the OR(95% CI)values were 2.475(1.172 6.521),1.597(1.12 2.764)respectively(P < 0.05).(2)Taken coronary artery disease as dependent variable(yes=1,no=0)and genotype(CC=0,CA=1,AA=2),TNF-α,IL-1,Hs-CRP,family history(yes=1,no=0),WC,BMI,HCY,Fbg,D-Dimer,TC,TG,HDL,LDL as independent variables to make Stepwise regression analysis,found that the genotype,HCY and LDL is closely related to the onset of coronary artery disease,the genotype and LDL can significantly increase the risk of coronary artery disease,they are independent risk factors for coronary artery diseasepathogenesis,the OR(95% CI)values were 9.576(1.753-52.311),1.728(1.112-4.327),2.237(1.013-7.453)respectively(P < 0.05).(3)Taken coronary artery disease as dependent variable(yes=1,no=0)and genotype(TT=0,TC=1,CC=2),TNF-α,IL-1,Hs-CRP,family history(yes=1,no=0),WC,BMI,HCY,Fbg,D-Dimer,TC,TG,HDL,LDL as independent variables to make Stepwise regression analysis,found that the genotype,LDL and D-Dimer is closely related to the onset of coronary artery disease,the genotype and LDL can significantly increase the risk of coronary artery disease,they are independent risk factors for coronary artery disease pathogenesis,the OR(95% CI)values were 4.494(1.398-22.292),2.26(1.314-5.13),2.582(1.545-12.218)respectively(P< 0.05).Summary:1 The increased serum concentration of TG,LDL,Glu,Fbg,D-D,HCY,HsCRP,CHI3L1,TNF-α,IL-1,GP6 and VCAM in Coronary artery disease group were closely related to the occurrence of coronary artery disease(CAD).2 The distribution of the gene polymorphism of SNP site rs4950928 in CHI3L1 gene in coronary artery disease group was obviously higher than that of normal control group,two groups exist significant differences,prompt that the gene polymorphism of SNP site rs4950928 was closely related to coronary artery disease,TT genotype significantly increased the risk of onset of coronary artery disease.3 The distribution of the gene polymorphism of SNP site rs1671153 in GP6 gene in coronary artery disease group was obviously higher than that of normal control group,two groups exist significant differences,prompt that the gene polymorphism of SNP site rs4950928 was closely related to coronary artery disease,AA genotype significantly increased the risk of onset of coronary artery disease.4 The distribution of the gene polymorphism of SNP site rs1654419 in GP6 gene in coronary artery disease group was obviously higher than that of normal control group,two groups exist significant differences,prompt that thegene polymorphism of SNP site rs4950928 was closely related to coronary artery disease,CC genotype significantly increased the risk of onset of coronary artery disease.5 Gene polymorphism of CHI3L1,GP6,LDL,HCY,D-Dimer were independent risk factors for coronary artery disease,and monitoring these factors might be valuable for prevention of coronary artery disease. |
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