| Part I: Distribution characteristics of PLCE1 gene single nucleotide polymorphisms in children of Guangxi Zhuang nationalityObjective: The distribution of single nucleotide polymorphism,rs10786156,rs3740360 and rs2274224 in children of Guangxi Zhuang was analyzed,and the genotype and allele frequency distribution differences in different ethnic groups and regions were compared。Methods: A total of 100 Guangxi Zhuang children who were undergoing physical examination in the Affiliated Hospital of Youjiang Medical College for Nationalities were selected to detect SNPs at four sites of PLCE1 gene rs17109674,rs10786156,rs3740360 and rs2274224 by next-generation sequencing.The Hardy-Weinberg Equilibrium(HWE)genetic law was used to evaluate whether the sample was population representative,SPSS22.0 statistical software was used to analyze the experimental data,and the genotype and allele frequency of the above points were compared with other ethnic and regional distribution differences in Guangxi Zhuang children by chi-square test or fisher exact probability method.Results: 1.Distribution of PLCE1 SNP locus genotypes and alleles in Guangxi Zhuang children:(1)G(63.5%)and A(36.5%)alleles and GG(36.0%),GA(55%)and AA(9.0%)genotypes were detected at rs17109674 locus of PLCE1 gene in Guangxi Zhuang children;rs10786156 loci detected G(63.5%),C(36.5%)alleles and GG(44.0%),GC(39.0%),CC(17.0%)genotypes;rs3740360 loci detected A(76.0%),C(24.0%)alleles and AA(60%),AC(32%),CC(7.0%)genotypes.The C(62.5%)and G(37.5%)alleles and CC(42.0%),CG(41.0%)and GG(17.0%)genotypes were detected at the rs2274224 locus.rs17109674,rs10786156,rs3740360 and rs2274224 genotypes of the four SNP loci of the PLCE1 gene The differences in the distribution of frequencies among healthy male and female Guangxi Zhuang children were not statistically significant(P>0.05).(2)The differences in the frequencies of the allelic genotypes of the four SNP loci of rs17109674,rs10786156,rs3740360 and rs2274224 in healthy male and female Guangxi Zhuang children were also not statistically significant(P>0.05).2.Distribution of PLCE1 gene SNP loci in different ethnic groups and regions:((1)rs17109674 genotype and allele frequencies were statistically significant(P<0.05)compared with those of the Beijing Han population and the Japanese population published in the 1000 Genomes Project database,but the differences were not statistically significant(P>0.05)compared with those of the Xishuangbanna Dai population;compared with the Finnish population,the differences in genotype frequency distribution were(P<0.05),and no statistically significant differences in allele distribution(P>0.05).(2)The differences in genotype and allele frequencies of rs10786156 loci were statistically significant(P<0.05)compared with those of the Beijing Han Chinese,Japanese and Finnish populations in the 1000 Genomes database,but not compared with those of the Xishuangbanna Dai population(P>0.05).(3)The genotypes and allele frequencies of rs3740360 loci were statistically significant(P<0.05)compared with those of the Beijing Han population and the Finnish population in the 1000 Genomes database,but not compared with those of the Xishuangbanna Dai population and the Japanese population(P>0.05).(4)The genotypes and allele frequencies of rs2274224 loci were statistically significant(P<0.05)compared with those of the Beijing Han population,Japanese population and Finnish population in the 1000 Genomes database,but not compared with those of the Xishuangbanna Dai population(P>0.05).Conclusion: The four SNPs of PLCE1 gene rs17109674,rs10786156,rs3740360 and rs2274224 in children of Guangxi Zhuang ethnic group were distributed differently in different regions and were regional.Part II Study of PLCE1 gene polymorphism and its serological level associated with primary nephrotic syndrome in Guangxi Zhuang childrenObjective: To investigate the correlation between four SNP loci of PLCE1 gene rs17109674,rs10786156,rs3740360,rs2274224 and the occurrence of primary nephrotic syndrome(PNS)and its response to hormone therapy in Guangxi Zhuang children,and also to study the expression of PLCε1(Phospholipase Cε1,PLCε1)protein serological level in Guangxi Zhuang children with PNS and analyze its significance.Methods: In a retrospective study,155 children with PNS in Guangxi Zhuang and 100 healthy children in Guangxi Zhuang during the same period were selected as study subjects.Four SNP loci of PLCE1 gene rs17109674,rs10786156,rs3740360,rs2274224 were genotyped using Fast Target combined with second-generation sequencing technology;meanwhile,the serum concentration of PLCε1 protein was detected by enzyme-linked immunosorbent assay(ELISA).ELISA was used to detect the serum concentration of PLCε1 protein in the samples.SPSS22.0 statistical software was used to analyze the experimental data,and the HWE genetic law was used to assess whether the healthy control samples were representative of the population,the chi-square test or Fisher’s exact probability method was used to analyze the differences in genotype and allele distribution,logistic regression was used to analyze the correlation between each SNP locus and the onset of PNS and its response to hormone treatment in Guangxi Zhuang children,and Mann-Whitney U rank sum test to analyze the differences in PLCε1 protein concentration levels between groups,and chain imbalance and haplotype analysis by SHEsis software.Results:(1)The differences in the distribution of rs3740360 locus genotype,dominant model(AC+CC vs AA),and allele frequency between the PNS group and healthy controls were statistically significant(X~2=11.725,P=0.03;X~2=8.884,P=0.003;X~2=5.770,P=0.016).rs3740360 locus dominant model AC+CC genotype,AC genotype and C allele were associated with the risk of PNS development,and after excluding confounding factors such as sex and age,it was suggested that AC+CC genotype,AC genotype and C allele were still associated with PNS development analysis(OR: 0.449,95% CI: 0.256-0.786,P=0.005;OR=0.354,95% CI : 0.188-0.667,P=0.001;OR=0.615,95% CI: 0.390-0.971,P=0.037).(2)There was no statistically significant difference in the distribution of genotypes and allele frequencies of rs17109674,rs10786156,and rs2274224 among the PNS and healthy groups(P>0.05).Further by logistic regression analysis with logistic regression correction after also excluding confounding factors such as sex and age,both results showed that the genotypes and alleles of the above three loci were not associated with the risk of developing PNS(P>0.05).(3)The differences in the distribution of genotype and allele frequencies of four loci,rs17109674,rs10786156,rs3740360,and rs2274224,between hormone-sensitive nephrotic syndrome(SSNS)and hormone-resistant nephrotic syndrome(SRNS)were not statistically significant(P>0.05).Further by logistic regression analysis with logistic regression correction after also excluding confounding factors such as sex and age,both results showed that the genotype and allele of the above four loci were not associated with the risk of SRNS development(P>0.05).(4)Four loci of PLCE1 gene rs17109674,rs10786156,rs3740360,and rs2274224 had linkage disequilibrium.(5)Haplotype results showed statistically significant differences in the distribution of AGCG,GGCC,and GGAG haplotypes between the PNS group and healthy controls(OR: 0.282,95% CI: 0.079-1.008,P=0.038;OR: 0.327,95% CI: 0.111-0.967,P=0.034;OR.4.616,95% CI 1.179-18.069,P=0.016),suggesting that AGCG and GGCC haplotypes may reduce the risk of developing PNS in Guangxi Zhuang children,while GGAG haplotype may increase the risk of developing PNS in Guangxi Zhuang children.(6)The serum PLCε1 protein level in the PNS group was significantly lower than that in the healthy group control group,and the difference was statistically significant(P<0.001);the PLCε1 protein level in the SRNS group was also lower than that in the SSNS group,and the difference was statistically significant(P<0.05).Conclusion:(1)The rs3740360 locus may be associated with the development of PNS in Guangxi Zhuang children,where the AC genotype,AC+CC genotype,and C allele may reduce the risk of developing PNS in Guangxi Zhuang children.(2)Four SNPs loci,rs17109674,rs10786156,rs3740360 and rs2274224,had linkage disequilibrium on PLCE1 gene;AGCG and GGCC haplotypes might reduce the risk of PNS development in Guangxi Zhuang children,while GGAG haplotype might increase the risk of PNS development in Guangxi Zhuang children.(3)Serum PLCε1 protein level may be a protective protein for PNS in Guangxi Zhuang children and may be associated with the response of PNS to hormone treatment in Guangxi Zhuang children. |
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