| Object: By observing the changes of biochemical markers and mi RNA expression in cerebrospinal fluid(CSF)of patients with cerebral hemorrhage,screening CSF markers for the diagnosis of cerebral hemorrhage diseases at the protein level and nucleic acid level,and then evaluate its clinical application value.Methods: Twenty-six patients with cerebral hemorrhage(ICH)disease were enrolled and divided into mild ICH group(13 cases)and severe ICH group(13 cases)according to Glasgow score(GCS).At the same time,22 hospitalized patients without brain diseases were selected as the control group.The CSF samples of each group of subjects were collected for 3~5ml within 48 hours after admission,and the samples were prepared by pre-treatment of the original samples according to the purpose and requirements of the experiment.At the protein level: The expression of ?2 microglobulin(?2-MG),aspartate aminotransferase(AST),lactic acid(LAC),lactate dehydrogenase(LD),microalbumin(m Alb),neuron-specific enolase(NSE)and astrocyte-derived protein(S100B)was detected by automatic biochemical analyzer and chemiluminescence immunoassay.Assess the diagnostic value of the above indicators in cerebral hemorrhage diseases;at the level of nucleic acid level: using cel mi RNA-39 as an external reference,q RT-PCR method was used to detect 15 target mi RNAs related to central nervous system diseases reported in the literature.The expression changes and Diagnostic value of each target mi RNA in cerebral hemorrhage diseases were observed and evaluated.Bioinformatics analysis is then used to predict target genes potentially associated with brain damage.Results: At the protein level: compared with the control group,there was no significant difference in the expression of LD,GLU and CL between the mild ICH group and the severe ICH group(P>0.05).The levels of m Alb,PRO,and albumin index(QAlb,QAlb=CAlb/SAlb)were significantly increased in the mild and severe ICH groups(P <0.05),but did not change with the degree of ICH injury;The levels of NSE,S100 B,?2-MG,AST and LAC were significantly increased in the light and severe ICH group(P<0.05).And with the degree of ICH damage showed an increasing trend.Based on the preliminary pre-experiment results and combined with the literature reports,S100 B and NSE were selected as the next research objects.The ROC curve analysis of the two showed that the AUC,sensitivity and specificity of NSE and S100 B diagnosis of ICH were 0.931 [95% confidence interval: 0.857~0.892],80.8%,95.5% and 0.795 [ 95% CI: 0.670~0.921],76.9%,77.3%;The combined detection of the two can improve the AUC of the diagnostic ICH to 0.965 and the sensitivity to 95.6%.Further studies showed that the level of NSE was significantly correlated with the amount of bleeding in ICH patients(r=0.86;P<0.05),and the level of S100 B was weakly correlated with the amount of bleeding in ICH patients(r=0.49;P<0.05);There was a positive correlation between NSE,S100 B and Qalb(r=0.443,0.606,respectively)(P< 0.05),suggesting that the two indicators are related to blood-brain barrier damage;NSE,S100 B and NIHSS scores were positively correlated(r=0.596,0.615,respectively)(P<0.05),suggesting that S100 B and NSE proteins can be used for evaluation of neurological deficits in cerebral hemorrhage.Furthermore,the ROC curve analysis of its diagnostic efficacy showed that the sensitivity of cerebral hemorrhage in the low and high NIHSS scores of S100 B and NSE was 78.9% and 80.8%,and the specificity was 73.3% and 55.2%,respectively.The sensitivity was significantly increased to 95.9%(P<0.05).At the level of nucleic acid level,the expression of CSF-mi R-124-3p and mi R-146a-5p was significantly up-regulated in ICH patients,and then their efficacy in diagnosing ICH was analyzed.The results showed that the AUC,sensitivity and specificity of mi R-124-3p and mi R-146a-5p for diagnosis of ICH were 0.667 [95% CI: 0.512~0.822],68.8%,81.8% and 0.878 [95% CI: 0.776,~0.844],50.1%,86.4% respectively.The combination of the two can improve the sensitivity of ICH diagnosis to 84.4%;Further bioinformatics analysis showed that mi R-124-3p can predict 91 target genes,mi R-146a-5p can predict 126 target genes,including SERP1,PDCD6,VEGFR-2.It is suggested that mi R-124-3p and mi R-146a-5p may be involved in apoptosis,inflammatory reaction,vascular endothelial injury and repair process.Conclusion: 1.NSE and S100 B in cerebrospinal fluid can be used as a laboratory auxiliary diagnostic index for cerebral hemorrhage,and the change of expression level can reflect the process of brain injury;2.The expression of mi R-124-3p and mi R-146a-5p in cerebrospinal fluid of patients with cerebral hemorrhage was significantly upregulated,and the combined detection of the two could be used for laboratory-assisted diagnosis of ICH;3.mi R-124-3p and mi R-146a-5p may be involved in the pathophysiological process of brain injury. |
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