| Purpose: 1.Establish expression profile of serum lncRNAs in children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis,and screen differentially expressed lncRNAs compared with healthy children;2.Analyze the enrichment functions and regulated signaling pathways of target genes of differentially expressed lncRNAs and explore the pathogenesis of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis;3.Explore the feasibility and clinical value of differentially expressed lncRNAs as potential diagnostic markers for children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.Material and method: This study adopted a case-control design in three stages.The first stage was the initial screening stage.A total of 10 samples of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis and healthy children were collected.High-throughput sequencing technology was used to detect expression profiles of lncRNAs in serum.The screening criteria was expression fold change FC?1.5 and P<0.05.The second phase was independent verification stage.30 children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis were collected respectively(three groups were set: isolited hematuria group;isolated proteinuria,hematuria and proteinuria group;moderate proteinuria group)and 20 healthy children in the control group.The five candidate lncRNAs screened in the initial screening stage were verified by RT-qPCR.The third stage was Pearson correlation coefficient analysis and ROC curve analysis were used to evaluate the value of single or multiple differentially expressed lncRNAs as potential clinical diagnostic markers for children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.Results: The expression profile of serum lncRNAs was obtained for the first time using high-throughput sequencing technology in children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.Compared with healthy children,820 differentially expressed lncRNAs were screened from the serum of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.34 differentially expressed lncRNAs were up-regulated and 786 differentially expressed lncRNAs were down-regulated.2.Bioinformatic technology was used to analyze differentially expressed lncRNAs.It was found that differentially expressed lncRNAs were involved in the production of proteins in vivo and participated in the regulation of Fas signaling pathway,p53 signaling pathway,NF-?B signaling pathway,B cell receptor signaling pathway and T cell receptor signaling pathway.3.The verification results of RT-qPCR of five lncRNAs with the most significant FC value(ENSG00000202354,ENSG00000252310,ENSG00000259001,HLA-B and ENSG0000025762)were consistent with the sequencing results in the expanded samples,which is statistically significant,indicating that the high-throughput sequencing results were relatively accurate.4.The results of Pearson correlation coefficient analysis showed that there were positive correlations between three lncRNAs and clinical grades of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis:ENSG00000252310(r=0.186,P<0.001),HLA-B(r=0.63,P<0.001),ENSG00000257621(r=0.42,P<0.001).The other two lncRNAs were negatively correlated with the clinical grade of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis: ENSG00000202354(r=-0.27,P<0.001),ENSG00000259001(r=-0.04,P<0.001).The r values of all the five lncRNAs ranged between(-1,1)and P values were statistically significant.5.The results of ROC curve analysis showed that the AUC of the five lncRNAs(ENSG00000202354,ENSG00000252310,ENSG00000259001,HLA-B and ENSG00000257621)ranged from 75.5% to 88.6%,both of which were statistical significance(P<0.001).The diagnostic efficiency of independent ENSG00000259001 as a diagnostic marker: AUC was 0.886,diagnostic index was 0.700,sensitivity was 1.000(0.832-1.000),specificity was 0.700(0.506-0.853),positive predictive rate was 0.690(0.492-0.847),negative diagnostic rate was 1.000(0.839-1.000),positive likelihood ratio was 3.330(2.600-4.200),and negative likelihood ratio was 0.The diagnostic efficiency of independent application of HLA-B as a diagnostic marker: AUC was 0.853,diagnostic index was 0.650,sensitivity was 0.750(0.509-0.913),specificity was 0.900(0.735-0.979),positive diagnostic rate was 0.833(0.586-0.964),negative diagnostic rate was 0.844(0.672-0.947),positive likelihood ratio was 7.500(5.700-9.900),and negative likelihood ratio was 0.280(0.070-1.000).6.Combined use of ENSG00000259001 and HLA-B for diagnostic efficiency detection showed that the combined AUC increased to 0.883(0.761-0.957)and the diagnostic sensitivity and specificity were 0.75 and 0.97,respectively.Conclusion: 1.Compared with healthy children,there was a significant difference in the expression of serum lncRNA in children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.2.Differentially expressed lncRNA may play an important role in the pathogenesis of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis by regulating Fas,p53,NF-?B,B cell receptor and T cell receptor signaling pathway.3.The results of Pearson correlation coefficient analysis showed that the diagnosis efficiency of HLA-B was the highest,suggesting that HLA-B could be used as a clinical marker to judge the severity of the condition of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.The greater the expression of HLA-B in the blood,the more severe the condition of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.4.The results of Pearson correlation coefficient analysis showed that ENSG00000257621 could be used as a secondary reference marker to determine the severity of childrensyndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.ENSG00000252310 and ENSG00000259001 were not suitable as clinical markers to determine the severity of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.5.The results of ROC curve analysis showed that HLA-B and ENSG00000259001 had high diagnostic efficiency.Combined with HLA-B and ENSG00000259001 could improve the sensitivity and specificity of diagnosis,suggesting that HLA-B combined with ENSG00000259001 could be used as a potential diagnostic marker for children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis.6.The results of ROC curve analysis showed that ENSG00000202354,ENSG00000252310 and ENSG00000257621 could be used as secondary reference markers for the diagnosis of children with syndrome of collateral injured by toxic and heat of henoch-sch?nlein purpura nephritis. |
PDF Full Text Request