Research On The Mutation And Expression Of NKX2.5Gene In Congenital Heart Disease With Diminutive Pulmonary Blood

Objective： Researching on the mutation and expression of NKX2.5gene incongenital heart disease with diminutive pulmonary blood, We preliminarily explorethe association between NKX2.5gene and pathogenesis of congenital heart diseasewith diminutive pulmonary blood,which provides the basis for the prevention andtreatment for congenital heart disease.Methods:Experimental group:56cases of peripheral venous blood and hypertrophicmyocardial tissue from right ventricular outflow tract when the operation isconventional resection and reservation are collected in congenital heart disease withdiminutive pulmonary blood,whose size is0.5*0.5*0.5cubic centimeter,which isfrom the cardiac surgery,where are the1st Affiliated Hospital of Bengbu MedicalCollege and Anhui Province Children Hospital in May2012to May2013, ranging inage from6months to14years old, with an average age of6.3years.Control group:63cases of peripheral venous blood, the right front wall outflow tract incision orabnormal muscle tissue from right ventricle, whose size is0.5*0.5*0.5cubiccentimeter,the specimens are from the cardiac surgery, where are the1st AffiliatedHospital of Bengbu Medical College and Anhui Province Children Hospital in May2012to May2013, ranging in age from6months to14years old, the average age of6.9years.Using Polymerase Chain Reaction combined with DNA sequencingtechnology to detect NKX2.5gene exon sequence, and analyse whether NKX2.5genemutation associates with Congenital Heart Disease with diminutive pulmonaryblood;Extraction of myocardial tissue RNA,taking GAPDH as internal control, is todetect the expression changes of NKX2.5gene mRNA and make a statistical analysisstatistical analysis in the two groups of myocardial tissues.Results: Firstly,using polymerase chain reaction combined with DNA sequencingtechnology, the PCR amplification product length is in conformity with the design of the fragment length, which specificity is also ideal.United States ABI3130geneticanalyzer was used to capillary electrophoresis after purifying product, sequencingresults show that the type does not appear abnormal peak compared with NKX2-5gene loci when96hole boards in accordance with the procedures are included in theanalyzer automatic sequencing,56cases of experimental group and63cases of controlgroup are not detected in the peripheral venous blood NKX2.5gene exonmutation.Secondly,Real-time fluorescent quantitative PCR method are used tocalculate experimental NKX2.5mRNA relative quantity that can be further calculatedthe relative quantitative myocardial NKX2.5gene mRNA expression in theexperimental group is0.5638±0.0051, shows that the experimental group NKX2.5inmyocardial mRNA expression level decreases, SPSS17.0software is detected theexpression level by t test that there are differences between two groups (p<0.01),which shows that the expression level of differences between two groups hasstatistical significance,With the method of times than dilution cDNA samplesamplifying GAPDH and NKX2.5gene, calculating to participate in the project withineach of the diluted sample dilution gradient value of the gene and Δ Ct, in cDNAlogarithm concentration curve of the relationship with Δ Ct, the slope is0.0488, closeto zero, namely the amplification efficiency is basic consistent with GAPDH andNKX2.5gene,according to the logarithmic concentration of cDNA with thecorresponding Ct value and NKX2.5,GAPDH corresponding correlation coefficientsare0.9987and0.9992, the slope is3.3728and3.4017respectively, amplificationefficiency of97%and98%respectively, NKX2.5GAPDH and has a good linearrelationship between them.Conclusion: The factor of NKX2.5gene mutation may be associated with multiple factors; theoccurrence of congenital heart disease with diminutive pulmonary blood may berelated with decline in the myocardial tissue NKX2.5gene expression.