| Objectives By detecting the left-right shunt congenital heart disease blood cells of TGF-β1gene polymorphism+869T/C expression levels of TGF-β1and left-right shunt congenital heart disease-related myocardial fibrosis, as well as gene+869T/C polymorphism in the significance of myocardial fibrosis.Methods Surgical treatment of randomly selected year or echocardiography confirmed that left-right shunt congenital heart disease (VSD, ASD, PDA) in children with43cases, including26males and17females, aged0.288years, mean1.43±2.65years. Matched normal healthy children in35cases, including23males and12females, aged0.39to7years, mean2.51±2.24years. To obtain informed consent of their parents early morning fasting venous blood, the blood cells to extract DNA, through the establishment of TGF-β1/β-actin real-time quantitative PCR assay for detection of clinical samples were compared between the two different alleles the frequency distribution of gene copy number analysis+869T/C gene copy number variation and myocardial fibrosis.Results (1) The quality and quantity of the total DNA:OD260/OD280was between1.7-1.8by UV spectrophotometer detection.(2) The gel electrophoresis showed the band240bp and the137bp band were clear.(3) The standard curve had good liner correlation, the congtiguous coefficient R2>0.99. The standard curve equation of TGF-β1+869T/C gene was Y=-0.2964X+7.8603, R2=0.999; β-actin gene was Y=-0.2888X+0.9989, R2=0.9989.(4) The respective ratio of+869/β-actin between the experimental group and the control group was0.0881±0.0397(n=43) and0.0436±0.0061(n=35), t=2.632(n=78,p<0.05). The difference was statistically significant.(5) Experimental group and control group three genotypes (TC, TT, CC) the actual and predicted values measured by the chi-square test p>0.05, the difference was not statistically significant.Conclusions (1) the use of real-time quantitative TaqMan probe confirmed that the human genome technology of TGF-β1gene in left-right congenital heart disease is highly expressed.(2) left-right shunt congenital heart disease group and normal control group genotypes TGF-β1+869gene locus T/C mutation in the left-right shunt congenital heart disease, myocardial fibrosis has important of significance.(3) The experimental use of the TaqMan probe real-time quantitative PCR with high specificity and high reliability, is the ideal method of detecting gene expression levels. |
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