| Objective:Hepatitis B caused by hepatitis B virus(HBV)infection has become one of the serious public health problems worldwide.People with HBV infection often become high-risk people with cirrhosis and liver cancer.The kidney,as one of the main pathogenic organs other than the liver,has gradually attracted attention in the pathogenesis of hepatitis B virus-associated glomerulonephritis(HBV-GN).Epidemiological studies have found that some HBV carriers have a tendency to involve the kidneys and develop nephritis.The Center had retrospectively analyzed the spectrum of renal pathological types and their trends in the past 20 years(1995-2015).The diagnosis of HBV-GN in nephropathy is on the rise.At present,HBV antigen or HBV-DNA is detected as an important diagnostic basis for HBV-GN in kidney tissue sections.However,many existing detection methods lack certain sensitivity and specificity,and it is always necessary to look for HBV-GN immunological markers.It is the focus of renal pathologists.Hepatitis B virus X gene is the fourth open reading frame of the double-stranded circular DNA molecule of HBV gene.Hepatitis B virus X protein(HBX protein)is the basic viral protein in the viral infection cycle in vivo,which can act on viruses and host cells in a direct and indirect manner.It is closely related to the occurrence and development of hepatitis B-associated nephritis.This study retrospectively analyzed the distribution of HBX protein in HBs Ag-positive kidney tissue,the relationship between HBV-DNA levels and clinical pathological types,and further explored the expression and significance of HBX protein and HBV-DNA levels in glomerulonephritis with hepatitis B virus,and provided a theoretical basis for finding reliable detection indicators in HBV-GN kidney tissue.Methods:1.Collect case and clinical data: 102 cases of renal biopsy of patients with HBV serological positive in our center from January 2013 to December 2015 were selected.The clinical and laboratory data of the patients were collected,including: patient gender,age at renal biopsy.And indicators related to HBV infection: HBs Ag,HBe Ag,HBs Ab,HBc Ab,HBe Ab.2.Analysis of clinical pathological types of 102 cases of renal biopsy: The renal tissue specimens were divided into three parts.Paraffin specimens were sliced in3μmthickness anddetected by light microscopy(HE,PAS,Masson,PASM staining),immunofluorescence(IF)and Electron microscopy(EM).Immunohistochemical method(IHC)was used to detect the expression of HBs Ag and HBc Ag in renal tissues,and the expression differences between different pathological types were analyzed.3.Expression of HBX protein in renal tissue: Immunohistochemical method(IHC)was used to detect the expression of HBX protein in renal tissue,and its expression difference between different pathological types was analyzed.4.Detection of HBV-DNA content in kidney tissue: The content of HBV-DNA was detected by Q-RT-PCR method,and the difference in expression between different pathological types was analyzed.5.Evaluation of three detection methods in the diagnostic value of HBV-GN:comparing the expression differences between HBX,HBs Ag,HBc Ag and HBV-DNA,combined with clinical data and pathological features,retrospective statistical analysis,and then evaluate its valueof thediagnosis of hepatitis B viral infection-associated nephritis.Results:1.Among the 102 patients with seropositive hepatitis B virus collected by our center,22 were patients with large three positive patients,3 of whom were in the DNA replication phase.The incidence of men is slightly more than that of women,and it is morecommon around 40 yearsold.2.Analysis of clinical pathological types and expression of HBs Ag and HBc Ag in102 cases of renal biopsy:(1)Pathological types showed diversity,including: HBV-MN(membrane hepatitis B associated nephritis)in 24 cases,UAMN(atypical membranous nephropathy)in 18 cases,IMN(idiopathic membranous nephropathy)in 24 cases,Ig AN(Ig A nephropathy)15 cases,MCD(glomerular micro lesions)in 18 cases,FSGS(focal segmental glomerulosclerosis)in 2 cases,HBV-MPGN(membrane proliferative hepatitis B associated nephritis)in 1 case.(2)The positive rates of HBs Ag and HBc Ag in HBV-MN group were 50% and62.5%,respectively.The positive rates of HBs Ag and HBc Ag in UAMN group were16.67% and 22.22%,respectively.The positive rates of HBs Ag and HBc Ag in IMN group were 12.50% and 33.33%,respectively.One patient with HBV-MPGN only expressed HBc Ag;the expression of HBs Ag and HBc Ag was not detected in Ig A group,MCD group and FSGS group.3.Expression of HBX protein in renal tissue of 102 serologically positive patients:(1)71 cases(69.6%)were positively expressed in brown granules deposited in glomerular podocytes,subepithelial capillary vasospasm and renal tubular epithelial cytoplasm.All positive cases were mainly expressed in renal tubular epithelial cells,accompanied by Different degrees of capillary vasospasm and podocyte expression.(2)The positive expression rate of HBX protein in 25 HBV-GN groups was 80.00%,and the positive expression rate of HBX-MN renal tissue was 83.33%(20/24 cases),which was higher than that of IMN group(50.00%)(12/24 cases);UAMN The positive expression rate of HBX was about 83.33%(15/18 cases),and the positive rates of Ig AN and MCD groups were 46.67%(7/15 cases),83.33%(15/18 cases),and 1 case of HBV-MPGN.Positive expression of HBX protein was detected,and positive expressionof HBX was detected in 2 patients with FSGS.(3)The positive expression rate of HBX protein in Dasanyang group was about81.82%(18/22 cases).There was no significant difference in the positive expression rate of HBX protein between the “Da Sanyang” group and the “Non-Sanyang” group.(4)The expression of HBX protein was detected in 3 cases of renal tissue during DNA replication.4.Detection of HBV-DNA in renal tissue of 102 serologically positive patients:(1)The positive expression of HBV-DNA in the renal tissues of 24 patients(23.53%)was detected by Q-RT-PCR.The positive rates of HBV-GN group and non-HBV-GN group were statistically significant(P<0.001).(2)The positive expression rate of HBV-DNA in the "Da Sanyang" group was45.45%(10/22 cases).(3)There was a significant difference in the positive expression rate of HBV-DNA between the "Da Sanyang" group and the "Non-Sanyang" group(P<0.05).(4)The expression of HBV-DNA was detected in all 3 kidney tissues during DNA replication.5.Evaluation of three detection methods in the diagnostic value of HBV-GN:(1)The positive expression rate of HBX protein in renal tissue was higher than that of HBs Ag and HBc Ag,and the difference was statistically significant in UAMN group(P<0.001),Ig AN group(P<0.001)and MCD group(P<0.001).(2)The positive expression rates of HBX,HBs Ag and HBc Ag in the renal tissues of25 HBV-GN patients were 83.33%,50.00% and 64.00%,respectively.There was no significant difference between the groups.(3)The positive expression rate of HBX protein in 102 cases of hepatitis B serone positive kidney tissue was higher than that of HBV-DNA,and the difference was statistically significant(P<0.001).Conclusion:1.Glomerulonephritis caused by HBV infection is one of the main pathological types of secondary glomerulonephritis in China,among which membranous nephropathy is the most common.HBX protein is mainly composed of renal tubular epithelial cytoplasmic deposition in renal tissue of patients with HBV serologically positive.2.The positive expression rate of HBX protein in HBS-positive patients was higher than that of HBs Ag and HBc Ag.The positive expression rate of HBX protein was higher than that of HBV-DNA,and the difference was statistically significant(P <0.001).The detection of HBX protein expression by immunohistochemistry can provide more evidence for pathological diagnosis of hepatitis B virus infection-associated nephritis. |
PDF Full Text Request