Diagnostic Value Of Serum Micrornas In The Progression Of Chronic Hepatitis B Fibrosis

Objective:This study aims to find out the differential expression of serum micro RNAs in the progression of chronic hepatitis B fibrosis and to explore its diagnostic value.Methods: A retrospective study was conducted.The cases were from patients with chronic hepatitis B infection who underwent liver biopsy from January 1,2017 to January 30,2019 in our hospital.Serum samples of healthy control group were collected from healthy people in our hospital physical examination center.A total of114 patients with chronic hepatitis B who had undergone liver puncture were enrolled.43 normal people came from the physical examination center of our hospital.Findings:3 healthy controls?CN?,3 mild liver fibrosis?MF?,3 moderate to severe liver fibrosis?SF?.Screening set: 20 healthy controls?CN?,20 mild liver fibrosis?MF?,20 moderate to severe liver fibrosis?SF?.Verification set: 20 healthy controls?CN?,36 mild liver fibrosis?MF?,and moderate to severe liver fibrosis.Thirty-two cases of severe hepatic fibrosis?SF?.After total RNA extraction and quality inspection,library construction,purification of the DNA,DNA cluster formation on Illumina's Cluster Station?Illumina q2000?and sequencing,the differentiated micro RNAs were detected by q RT-PCR from screening sets?20 cases for CN,MF and SF respectively?,and the expression level of micro RNAs was detected by Ct.Value measurement.The comparison of differentially expressed genes between the two groups was expressed by 2-Delta ct,and the micro RNA panel equation was established by logistic regression.The validation set was used to validate the diagnostic effect of micro RNA panel.Results: There was no significant difference in virological indexes between mild hepatic fibrosis group and moderate hepatic fibrosis group,but there was difference in ALT of liver function.Sequencing analysis showed that 97 39507,100 31720 and 9990867 primitive sequences were obtained after primary analysis in control group,mild and moderate chronic hepatitis B fibrosis group,and the remaining 289819,469367 and 31364 comparable sequences were obtained after redundancy removal.Comparing the three data after normalization,the expression of serum micro RNAs in CN and MF was different in different liver fibrosis injury.A total of 84 differentially significant micro RNAs were found,which accorded with the expression difference multiple of more than four times.Eight micro RNAs with P < 0.05 were up-regulated genes and 18 down-regulated micro RNAs.Serum micro RNAs were differentially expressed in both MF and SF.A total of 124 differentially significant micro RNAs were found,which accorded with the expression difference multiple more than 4times.Fifteen micro RNAs with P < 0.05 were up-regulated genes and 13down-regulated genes.The 47 differentially expressed micro RNAs were further screened by screening sample set.The screening conditions were as follows: CT value35,detection rate 75%.Four candidate micro RNAs were screened for expression difference between CN and MF,respectively: hsa-mir-34a-5p,hsa-mir-1228-3p,hsa-mir-122-5p and hsa-mir-26a-5p.Two candidate micro RNAs,hsa-mir-34a-5p and hsa-mir-337-5p,were screened for their differences in expression between MF and SF. Logistic regression was used to establish regression equation.MF differentiated Logiit P1=-3.354+0.9354* Mi₃4-0.3546* Mi₂6a of healthy control group.SF and MF were regarded as independent variables,and the circulation number of mi RNA was regarded as covariate.Logistic regression was used to establish regression equation.SF differentiates MF from Logit P2 =-4.235 + 1.2433 * Mi₃4a + 0.5483 *Mi₃37.The area under the micro RNA panel 1?MF vs health?curve?AUC?was0.829?95% CI = 0.705 to 0.917?;sensitivity = 66.7%,specificity = 90%?P < 0.0001?.The AUC of micro RNA panel 2?SF vs MF?was 0.663?95% CI = 0.518 to 0.788?;sensitivity = 50%,specificity = 85%?P < 0.05?.Conclusion: Micro RNAs play an important role in the progression of chronic hepatitis B fibrosis.We found that there were differences in the expression of micro RNAs between normal controls and chronic hepatitis B fibrosis,and there were still differences in the expression of micro RNAs in the progression of hepatic fibrosis.Moreover,the combination of micro RNAs is valuable for the diagnosis of liver fibrosis.