| Objective:To detect the dynamic expression levels of HBV pre genomic RNA(pgRNA)and hepatitis B core associated antigen(HBcrAg)in circulating blood of patients with chronic hepatitis B(CHB)before and after treatment.The expression levels of HBV pgRNA and HBcrAg in circulating blood of patients with CHB and their correlation with the degree of disease were analyzed.To explore the relationship between the expression levels of HBV pgRNA and HBcrAg and the levels of HBV-DNA load,HBeAg and HBsAg.Objective to understand the change trend of HBV pgRNA and HBcrAg expression levels in patients with CHB during anti HBV treatment and after withdrawal.To investigate the expression of HBV pgRNA and HBcrAg in circulating blood of patients with CHB and their correlation with clinical value.To analyze the preliminary value of HBV pgRNA and HBcrAg in predicting the complete response of CHB patients after anti HBV treatment.Methods:Collected from March 2020 to October 2021 in affiliated hospital of North Sichuan Medical College outpatient department or inpatient department for the first time to use Entecavir(ETV)anti hepatitis B virus and other comprehensive treatment and followed up for 48 weeks in 105 cases of CHB patients as the treatment group.In the same period,39 cases of hepatitis B virus carriers,82 cases of hepatitis B related cirrhosis(compensatory stage)and 25 cases of hepatitis B related chronic subacute liver failure were selected as the control group.3ml of abdominal peripheral venous blood was collected from patients with CHB at the time of initial treatment,12 weeks after treatment,24weeks after treatment and 36 weeks after treatment,place it in heparin anticoagulant tube,centrifuge at 1000 rpm within 6 hours,and retain its supernatant after 15 minutes,and store it in-80℃ refrigerator.127 CHB patients who had been treated with anti HBV for 5-6 years(negative conversion of HBV DNA and HBeAg)and stopped taking drugs after informed consent in the Affiliated Hospital of North Sichuan Medical College,Suining Central Hospital,Mianyang 404 hospital and Nanchong central hospital were collected,including 10 patients who withdrew from follow-up due to liver damage and 117 CHB patients who were followed up for 24 weeks.HBV pgRNA in circulating serum of patients with chronic hepatitis B was detected by Real-time PCR.The expression levels of HBcrAg in peripheral venous blood of patients with chronic hepatitis B were detected by enzyme-linked immunosorbent assay(ELISA),HBV-DNA load with high accuracy was detected by real-time fluorescence quantitative method.The levels of HBsAg and HBeAg of HBV infection markers were detected by chemiluminescence.The test method of each index of the subjects in the control group is the same as that of the patients in the CHB group,which shall be operated by a specially assigned person within the effective use period according to the reagent operation manual.This study was approved by the ethics committee of the Affiliated Hospital of North Sichuan Medical College(File Number:2022ER 127-1).Results:1.With the progress of the disease,the expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA load in circulating serum decreased;The expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA load in circulating serum of CHB patients were lower than those in carrier group(P<0.05);Compared with cirrhosis group and chronic subacute liver failure group,the expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA in circulating serum of CHB patients were higher(P<0.05);There were significant differences in the levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA load in the circulating serum of the subjects(F=13.746,P=0.000;F=98.644,P=0.000;F=035,P=0.000;F=89.390,P=0.000).2.The expression levels of HBV pgRNA and HBcrAg in circulating serum of CHB patients decreased gradually with the aggravation of the disease.The expression levels of HBV pgRNA and HBcrAg in circulating serum of CHB patients were significantly different among the three groups(F=32.692,P=0.000;F=9.162,P=0.000).The expression level of HBV pgRNA and HBcrAg in circulating serum of mild CHB group was higher than that of moderate CHB group and severe CHB group(P<0.05);The expression levels of HBV pgRNA and HBcrAg in circulating serum were negatively correlated with the severity of CHB patients(F=-0.796,P=0.000;F=-0.670,P=0.000).3.After the treatment,the expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA load in circulating blood of patients with chronic hepatitis B decreased significantly,and the difference was statistically significant(P<0.05).4.The expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV DNA load in CHB patients were significantly lower than those before treatment at the 12th,24th and 48th week of ETV treatment(P<0.05),the difference was statistically significant;the expression levels of HBV pgRNA,HBcrAg,HBsAg and HBV-DNA in circulating serum of CHB patients were statistically significant at each period of treatment(F=12.278,P=0.000;F=23.799,P=0.000;F=120.501,P=0.000;F=576.577,P=0.000).5.Before treatment,the level of HBV pgRNA in HBeAg(+)group was higher than that in HBeAg(-)group(P<0.05),and the level of HBcrAg in HBeAg(+)group was higher than that in HBeAg(-)group(P>0.05);The expression levels of serum HBV pgRNA and HBcrAg in HBeAg(+)and HBeAg(-)groups before treatment were significantly higher than those in different periods after treatment(F=63.803,P=0.000;F=10.292,P=0.000;F=4.468,P=0.000;F=32.903,P=0.000).At the 48th week of treatment,the serum HBV RNA CHB patients in HBeAg(+)group were significantly lower than those before treatment(P<0.05).At the 48th week of treatment,there was no significant difference in the levels of HBV RNA and HBcrAg between HBeAg(+)and HBeAg(-)groups(P>0.05).6.There was a positive correlation between serum HBV RNA level and HBcrAg,HBsAg and HBV DNA load in CHB patients before treatment(r=0.620,P=0.000;r=0.729,P=0.000;r=0.720,P=0.000);There was a positive correlation between serum HBcrAg level and HBsAg and HBV DNA load(r=0.880,P=0.000;r=0.694,P=0.000).After treatment,there was a positive correlation between serum HBV RNA level and HBcrAg,HBsAg and HBV DNA load in CHB patients(r=0.550,P=0.000;r=0.770,P=0.000;r=0.551,P=0.000);There was a positive correlation between serum HBcrAg level and HBsAg and HBV DNA load(r=0.599,P=0.000;r=0.535,P=0.000).7.The predictive value of HBV pgRNA,HBcrAg,HBsAg and HBV DNA in the diagnosis of complete response in CHB patients was analyzed.The area under the curve(AUC)was obtained through ROC curve analysis.The results showed that the AUC(0.878)of HBV pgRNA was significantly higher than that of HBcrAg(0.736),HBV DNA(0.716)and HBsAg(0.662),P<0.05,The sensitivity and specificity were 0.83,0.86,0.75 and 0.66;0.85、0.58、0.73、0.62;The cut off values are 1.30,2.22,2.62 and 2.83.8.127 patients with CHB were treated with anti HBV for 5-6 years(negative conversion of HBV DNA and HBeAg)and stopped the drug without liver damage after informed consent,including 117 patients were followed up for 24 weeks.The expression levels of HBV pgRNA,HBcrAg and HBV DNA in circulating blood after drug withdrawal showed an upward trend.At 24 weeks after drug withdrawal,the expression level of HBV pgRNA and HBcrAg in circulating serum of CHB patients was significantly higher than that at the time of drug withdrawal and 4 weeks after drug withdrawal(P<0.05);At 24 weeks after drug withdrawal,the expression level of HBV DNA in circulating serum of CHB patients was significantly higher than that at the time of drug withdrawal(P<0.05).There were significant differences in the expression levels of HBV pgRNA,HBcrAg and HBV DNA in circulating serum of CHB patients among the four groups at 4,12 and 24 weeks after drug withdrawal(F=3.262,P=0.03;F=4.117,P=0.011,F=3.753,P=0.017).9.Among 127 CHB patients,21 cases recurred within 12 weeks and 18 cases recurred within 12-24 weeks after anti HBV treatment for 5-6 years.Among the recurrent patients,10 cases had abnormal liver function(HBV DNA and HBeAg were repositive alone or at the same time),including 2 cases of liver failure.There were 117 patients with CHB who stopped the drug without liver damage and were followed up for 24 weeks.At the time of withdrawal,the levels of HBV pgRNA and HBcrAg in recurrent patients were higher than that in the non recurrent patients(t=6.771,P=0.000;t=8.537,P=0.000).The level of serum HBV markers at 24 weeks was selected in the non relapsed group,and the level of serum HBV markers at relapse was selected in the relapsed group.The levels of HBV pgRNA,HBcrAg,HBV DNA and HBsAg in the circulating serum of relapsed patients were higher than those in the non relapsed group.The levels of HBV pgRNA,HBcrAg and HBV DNA,HBsAg in recurrent patients were significantly higher than those in non recurrent patients(t=11.045,P=0.000;t=15.871,P=0.032;t=19.209,P=0.000;t=20.174,P=0.000).Conclusion:1.The expression levels of HBV pgRNA and HBcrAg in circulating serum decreased with the aggravation of CHB patients.2.After treatment,the expression levels of HBV pgRNA and HBcrAg in circulating serum of CHB patients decreased significantly.With the extension of treatment time,the decline rate gradually slowed down.The expression level of HBV pgRNA was positively correlated with the level of HBcrAg and closely related to the level of HBsAg and HBV DNA load.3.The predictive value of HBV pgRNA and HBcrAg in circulating blood alone for complete response in patients with CHB showed that the sensitivity and specificity of HBV pgRNA and HBcrAg were higher than those of HBsAg and HBV DNA.4.At the time of withdrawal,the levels of HBV pgRNA and HBcrAg in recurrent patients were higher than that in the non recurrent patients,the detection of HBV pgRNA and HBcrAg levels may be one of the potential reference indexes for the endpoint selection of anti HBV treatment. |
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