Part One The Clinical Value Of FibroScan Combined With Hepatic Fibrosis Score In Non-invasive Diagnosis Of NASH Fibrosis

Part one The clinical value of FibroScan combined with hepatic fibrosis score in non-invasive diagnosis of NASH fibrosisObjective:To explore the clinical value of FibroScan combined with non-invasive hepatic fibrosis score in distinguishing different degrees of fibrosis in patients with NASH.Methods:1.41 NASH patients diagnosed by liver biopsy were selected from the Department of Geriatrics Gastroenterology,the first affiliated Hospital of Kunming Medical University from November 2018 to December 2020.The sex,age,height,weight,BMI,drinking history,diabetes history,hypertension history,PLT,blood biochemical indexes,CAP and LSM were collected,and the non-invasive liver fibrosis scores(FIB-4,NFS,APRI,GPR)were calculated.2.According to the liver histopathological score,the patients with NASH were divided into three groups according to different stages of liver fibrosis(S),grade of inflammation(G)and degree of steatosis(F).3.The differences of clinical indexes,CAP,LSM and non-invasive liver fibrosis scores among different groups were analyzed by SPSS26.0 software,and the correlation was analyzed by Spearman correlation.The joint predictors were constructed by multi-factor binary Logistic regression,and the ROC curves were drawn.The area under the curve(AUC)was calculated to evaluate the value of CAP,LSM and FIB-4,NFS,APRI,GPR in the diagnosis of different degrees of hepatic fibrosis,inflammation and steatosis in NASH.The pairwise comparison of AUC was performed by MedCalc18.2.1 software Delong method to evaluate the early warning value of various non-invasive diagnostic methods alone or in combination in the diagnosis of advanced liver fibrosis,moderate and severe inflammation and moderate and severe steatosis in NASH.Results:1.There is a high correlation between LSM and the fibrosis stage(S)in the pathological diagnosis of liver puncture(r=0.614).The highest area under the ROC curve of NASH patients is 0.889,which is higher than FIB-4,NFS,APRI and GPR.The diagnostic ability of AUC is better than APRI,GPR(P<0.05).There is no significant difference with FIB-4 and NFS,the highest diagnostic accuracy is LSM of 84.5%.2.Joint diagnosis:the highest area under the ROC curve of LSM combined with FIB-4 and NFS in the diagnosis of progressive fibrosis of NASH patients is 0.902,and there is no significant difference compared with other combined indexes of AUC(P<0.05).The accuracy of LSM combined with FIB-4,NFS,APRI and GPR in the diagnosis of progressive liver fibrosis of NASH is 87.25%,followed by FIB-4 or APRI,and the diagnostic accuracy is 86.2%.3.There is a high correlation between NFS and inflammatory grade(G)in the pathological diagnosis of liver biopsy(r=0.584).The highest area under the ROC curve to distinguish moderate and severe inflammation in NASH patients is 0.871(P<0.05),but it’s diagnostic accuracy is 82.75%the highest of others.The diagnostic accuracy of LSM and liver biopsy is 78.6%,second only to NFS.4.There is a high correlation between CAP and the stage of steatosis(F)in the pathological diagnosis of liver biopsy(r=0.533).The highest area under the ROC curve to distinguish moderate and severe steatosis in NASH patients is 0.847(P<0.05),the diagnostic accuracy is the highest(80.9%).CAP combined with APRI can improve the diagnostic value of NASH steatosis,and the diagnostic accuracy is 83.35%,which is higher than that of CAP alone.Conclusions:1.LSM combined with FIB-4 or APRI can better evaluate the progress of NASH fibrosis.2.CAP combined with APRI can more effectively and noninvasively evaluate the degree of steatosis in NASH.NFS can early identify moderate and severe inflammation in NASH.The diagnostic accuracy of NFS is higher than LSM in mild inflammation.3.The application of LSM,CAP and NFS can make a more comprehensive noninvasive diagnosis and evaluation of NASH patients.Part two The clinical significance of miR-6763-5p in the diagnosis of NASHObjective:To explore the diagnostic value of miR-6763-5p in NASH patients by detecting the expression of miR-6763-5p in plasma.Methods:1.46 patients with NASH and 40 healthy controls were selected from the Department of Geriatric Gastroenterology,the first affiliated Hospital of Kunming Medical University from January 2019 to December 2019.All the subjects were examined by FibroScan and divided into two groups according to CAP and LSM.2.Collected the sex,age,height,weight,BMI,drinking history,diabetes history,hypertension history,PLT,blood biochemical index,CAP and LSM of all subjects,and calculate the non-invasive liver fibrosis score(FIB-4,NFS,APRI,GPR).3.The fasting blood of all subjects was collected,and the expression level of miR-6763-5p in plasma was detected by RT-qPCR.4.The relative expression of miR-6763-5p in plasma and clinical related indexes were analyzed by SPSS26.0 software,and the expression picture of miR-6763-5p was drawn by GraphPad Prism8.0 software.A multivariate diagnostic model was constructed by binary Logistic regression,and the ROC curve was drawn to evaluate the ability of miR-6763-5p,non-invasive hepatic fibrosis score and multivariate diagnostic model to diagnose NASH.Results:1.Compared with CON,TP increased(P<0.05),body weight,BMI,ALT,AST,ChE,GGT,UA,TG,FBG,CAP,LSM,APRI and GPR increased significantly,while HDL-C decreased significantly in NASH group.The expression of miR-6763-5p in plasma of NASH is significantly lower than that of control group(P<0.01).3.Correlation analysis between clinical parameters and miR-6763-5p:there is a negative correlation between ALT,AST,GGT,CAP,LSM,GPR and the expression of miR-6763-5p,the highest correlation coefficient of GGT is-0.413,followed by GPR and ALT,correlation coefficient are-0.382 and-0.374 respectively.There is a positive correlation between the expression of HDL-C and miR-6763-5p,and the correlation coefficient is 0.221(P<0.05).3.Correlation analysis between clinical parameters and NASH:body weight,BMI,TP,ALT,AST,ChE,GGT,UA,TG,FBG,CAP,LSM,APRI,GPR are positively correlated with NASH,and the correlation coefficients are 0.455,0.532,0.234,0.479,0.369,0.369,0.598,0.394,0.536,0.378,0.864,0.781,0.292,0.590 respectively.There is a negative correlation between HDL-C,miR-6763-5p and NASH,and the correlation coefficients are-0.383 and-0.440 respectively.4.The AUC of miR-6763-5p for the diagnosis of NASH in normal population is 0.755(95%CI:0.651-0.859).It’s sensitivity,specificity,positive predictive value,negative predictive value,Youden index and diagnostic accuracy are 56.50%,92.5%,88.28%,68.01%,0.49 and 74.5%respectively.5.The AUC of FIB-4,NFS,APRI and GPR in the diagnosis of NASH are 0.524,0.534,0.669 and 0.841 respectively,among which FIB-4 and NFS have no diagnostic value(P<0.05),APRI,GPR have diagnostic value(P<0.01).6.A multivariate diagnostic model is constructed by Logistic regression.MiR-6763-5p,BMI,TG,UA and FBG,logistic(P)=-1.112*serum miR-6763-5p+0.391*BMI+1.78*TG+0.02*UA+0.952FBG-22.899 are included in the model.The AUC for the diagnosis of NASH is 0.965(95%CI:0.934-0.996).It is significantly better than the AUC of NASH diagnosed by miR-6763-5p,APRI and GPR alone.7.There is no significant difference in the expression of miR-6763-5p in plasma between NASH patients with different degrees of hepatic fibrosis and steatosis,and it is not possible to distinguish between different degrees of hepatic fibrosis and steatosis in NASH patients.Conclusions:1.The relative expression of miR-6763-5p is significantly decreased in patients with NASH.2.Constructing a five-variable joint diagnosis model with miR-6763-5p,BMI,TG,UA and FBG by logistic regression can significantly improve the diagnostic accuracy of NASH.3.MiR-6763-5p is expected to become a biomarker for the diagnosis of NASH.