Study On The Variation Sites Of The Whole Exogenous Group In The Syndrome Of Purpuric Nephritis With Heat Toxin And Collateral Injury

Purpose:1.Use high-throughput sequencing technology to search for the information of mutation sites in the whole exome of children with purpura nephritis and heat injury syndrome,and to screen for pathogenic mutation sites or mutation genes;2.Retrospective literature research,through the review and verification of candidate genes in NCBI,to determine the possible pathogenic genes of children with purpura nephritis and heat injury syndrome.Data and method:The subjects of this experiment consisted of 6 children with HSPN and 6 healthy children.The two-stage case-control design was adopted.In the first stage,DNA was extracted from the peripheral venous blood,and the entire exon region DNA of the sample was collected.Perform efficient enrichment and high-throughput sequencing.Bioinformatics analysis was performed on the obtained sequencing sequence,and finally the candidate disease-causing mutations related to the disease were obtained.In the second stage,through consulting a large number of literatures related to candidate genes,and searching for candidate genes in NCBI,the final identification of possible pathogenic genes.Results:1.Exome can be obtained is point mutations,that is,single-nucleotide polymorphism(SNP)changes and insertion The deletion(INDEL,insertion-deletion)mutation is insensitive to copy number variation(CNV).2.There were a total of 51141 SNP mutation sites in the 12 samples.Among them,SIFT,Polyphen,Mutation Taster,and CADD evaluated the harmfulness as 3999;there were 91211 INDEL mutation sites with 715 harmfulness.Refer to the evidence of the American Society of Medical Genetics and Genomics(ACMG)to classify the harmfulness of mutation sites,and the number of mutation sites classified as pathogenic and possible pathogenic is 62,and the number of uncertain mutation harmful sites is 7862 Pcs.3.On the basis of filtering harmful sites,there are 142 mutant genes and 246 mutation sites in the screening sample.4.Enrichment analysis of candidate genes revealed that they are involved in protein digestion and absorption,in the catalytic metabolism of single small molecules in cells,and in the regulation of p53 signaling pathway and TGF-β signaling pathway.5.Enter the disease name in Phenolyzer software and sort the candidate genes.The higher the ranking,the greater the correlation with the disease,and finally 19 possible pathogenic genes will be obtained.6.Check 19 pathogenic genes and related articles in NCBI,and HSPN related articles in Pub Med,and find the possible pathogenic genes for children with purpuric nephritis and heat injury syndrome are COL4A3 and COL4A4.Conclusion:1.Compared with the healthy group,there is a significant difference in the gene expression of the full-explicit group of children with purpura nephritis and heat injury syndrome;2.The differentially expressed pan-explicit mutation sites may participate in the catalytic metabolism process of a single small molecule in the cell and participate in the regulation of p53 signaling pathway and TGF-β signaling pathway;3.COL4A3 and COL4A4 may be the causative genes of children with purpura nephritis and heat injury syndrome,but the exact conclusion still needs to be verified by a large number of subsequent samples.