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Objective:To study the neuroprotective effect of Chuanjin Jieyu decoction on a chronic unpredictable stress (CUMS) induced depression rat model.Methods:A total of 48 adult male healthy Sprague-Dawley (SD) rats were randomly selected, with 12 rats as the control group, and the remaining rats were used to establish a CUMS depression model. After successful modeling, the rats were randomly divided into three groups: CUMS group, fluoxetine group, and Chuanjin Jieyu decoction group; 0.9% sodium chloride solution fluoxetine and Chuanjin Jieyu decoction were respectively administered for intervention for 4 weeks. After modeling and intervention, depression levels in rats were measured through open field experiments and sugar water preference experiments; After intervention, real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of brain-derived neurotrophic factor (BDNF), tyrosine protein kinase receptor B (TrkB), and cyclic adenosine monophosphate responsive element binding protein (CREB) mRNA in the dentate gyrus of the hippocampus. Immunofluorescence was used to detect the expression levels of BDNF and NeuN. Western blot was used to detect the level of glial fibrillary acidic protein (GFAP).Results:After modeling, the sugar consumption, sugar preference, vertical and horizontal movement results of the CUMS group, fluoxetine group, and Chuanjin Jieyu decoction group were significantly lower than those of the control group (all P<0.01); After intervention, the above indicators in the CUMS group were lower than those in the control group (all P<0.01). The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were higher than those in the CUMS group ( P<0.01 or P<0.05), but lower than those in the control group ( P<0.01 or P<0.05). The qRT-PCR results showed that the mRNA expression levels of BDNF, TrkB, and CREB in the CUMS group, the fluoxetine group, and the Chuanjin Jieyu decoction group were significantly lower than those in the control group (all P<0.01); The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were significantly higher than those in the CUMS group (all P<0.01). The immunofluorescence results showed that the expression levels of BDNF and NeuN in the CUMS group were significantly lower than those in the control group (all P<0.01); The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were higher than those in the CUMS group ( P<0.01 or P<0.05), and all indicators in the Chuanjin Jieyu decoction group and the NeuN expression level in the fluoxetine group were lower than those in the control group ( P<0.01 or P<0.05). Western blot results showed that the expression of GFAP protein in the CUMS group was lower than that in the control group ( P<0.01); The expression of GFAP protein in the fluoxetine group was higher than that in the CUMS group ( P<0.05). Conclusions:Chuanjin Jieyu decoction can improve the behavioral indicators of depression in rats, increase the expression of BDNF, TrkB, CREB, and NeuN in the hippocampal dentate gyrus, suggesting that Chuanjin Jieyu decoction may exert neuroprotective effects through the BDNF/TrkB/CREB pathway.
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Objective:We conducted a real-world multi-center clinical study with a large sample size to comprehensively evaluate the performance of three commercial hepatitis C virus (HCV) core antigen assays. The study aimed to evaluate the performance for their use in HCV infection screening, and to provide clues for further improving the sensitivity and specificity of the assays.Methods:Key performance indicators including the lower limit of detection (LOD), diagnostic sensitivity, and specificity of three HCV antigen assays (the Architect, Laibo, and ChemClin HCV core antigen assays) were evaluated using commercial seroconversion panels reflecting early HCV infection and clinical routine serum samples of outpatients and inpatients from 3 tertiary hospitals from January 2018 to April 2022. Factors that affect the performance indicators were further investigated.Results:The window period for detecting HCV infection with the three antigen assays was equal to or slightly longer than that of the RNA assay, but all are shorter than that of the anti-HCV assay. There was a good linear positive correlation between HCV core antigen and HCV RNA levels in treatment naive patients with hepatitis C ( r=0.90, P<0.01). For the most common genotype 1b strain in China, the LOD of the three HCV assays were equivalent to 531 IU/ml (Architect), 3,698 IU/mL (Laibo), and 4,624 IU/mL (ChemClin) HCV RNA, respectively. Due to the skewed distribution of HCV RNA levels in treatment-naive hepatitis C patients, more than 95% of the patients had viral loads higher than 6 166 IU/ml. Therefore, the three HCV antigens assays still maintained a satisfactory diagnostic sensitivity (94.33%-99.40%). Among 54 immunodeficient patients (leukemia patients) with HCV infection, 9% (5/54) had negative anti-HCV results, while the HCV antigen assays found all these infectors. Through further experiments, we revealed the amino acid polymorphism in the core region of genotype 3 strain impaired the sensitivity of all three HCV antigen assays. In addition, the sensitivity of the two domestic assays was impaired by anti-HCV antibodies in the serum. The specificity of HCV antigen assays for diagnosing hepatitis C is 99.94% to 99.98%. The rheumatoid factors, autoantibodies, and other unknown interference substances can lead to a small number of low level, "false positive" antigen results. Conclusions:HCV core antigen assay may be used as a satisfactory approach of infection screening, especially for the immunodeficient patents. However, the sensitivity and specificity of the assays are influenced by multiple factors, which should be further improved.
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【Objective】 To evaluate the role of anti-HBc detection in current blood screening strategy by the follow-up of repeated donors with antibody to hepatitis B virus core antigen. 【Methods】 Plasma samples were collected randomly from Dalian Blood Center. to test anti-HBc(dual reagents) and anti-HBs via ELISA. The re-donation of eligible donors who were anti-HBc+ and donors reactive to HBV detection were followed up. 【Results】 A total of 1 291 plasma samples were collected randomly from May 2017 to March 2018, among which 405 samples(31.4%)were anti-HBc+. The median age of anti-HBc+ group was observed much higher than that of anti-HBc-group (39 vs 31 years old) (P0.05). Among the 405 anti-HBc+ donors, 3 donors were OBI (0.7%), of which one was screened out in second donation. No HBV DNA was detected out in 3 OBI cases. 【Conclusion】 Although anti-HBc detection is not suitable in blood screening currently, it is of great value in the assessment of blood donor re-entry for HBV reactive donors in blood screening due to the high anti-HBc prevalence among blood donors.
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PCR is currently the non-debatable proof for diagnosis of HCV infection as well as conclusion of treatment outcomes. HCV core antigen (HCVcAg) testing is a neglected, less expensive and less time consuming test that's presumed to achieve the same aims. The aim of this study is to find the cost-effectiveness of HCV core antigen testing in the monitoring of treatment response as an alternative to the gold-standard PCR test
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Humans , Seroepidemiologic Studies , Environmental Monitoring , Public HealthABSTRACT
Abstract INTRODUCTION: We compared the hepatitis C virus (HCV) core antigen test with the HCV RNA assay to confirm anti-HCV results to determine whether the HCV core antigen test could be used as an alternative confirmatory test to the HCV RNA test. METHODS: Sera from 156 patients were analyzed for anti-HCV and HCV core antigen using a chemiluminescent microparticle immunoassay (Architect i2000SR) and for HCV RNA using the artus HCV RG RT-PCR Kit (QIAGEN) in a Rotor-Gene Q instrument. RESULTS: The diagnostic sensitivity, specificity, and positive and negative predictive values of the HCV core antigen assay compared to the HCV RNA test were 77.35%, 100%, 100%, and 89.38%, respectively. HCV core antigen levels showed a good correlation with those from HCV RNA quantification (r =0.872). However, 13 samples with a viral load of less than 4000 IU/mL were negative in the HCV core antigen assay. All gray-zone reactive samples were also RNA positive and were positive on repeat testing. CONCLUSIONS: The Architect HCV core antigen assay is highly specific and has an excellent positive predictive value. At the present level of sensitivity (77%), the study is still relevant in a low-income setting in which most of the HCV-positive patients would go undiagnosed, since HCV RNA testing is not available and/or not affordable. HCV core antigen testing can also help determine the true burden of infection in a population, considering the fact that almost 50% of the anti-HCV positive cases are negative for HCV RNA.
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Humans , Hepatitis C/diagnosis , Hepacivirus/genetics , RNA, Viral , Sensitivity and Specificity , Hepatitis C Antigens , Hepatitis C AntibodiesABSTRACT
Hepatitis B virus (HBV) belongs to the Hepadnaviridae family and can cause acute and chronic hepatitis, liver cirrhosis, and even liver cancer in humans. Current antiviral drugs cannot completely eliminate HBV in liver cells and thus it is difficult to achieve a curative effect. In recent years, the mechanism of persistent HBV infection has attracted wide attention, which mainly involves host and virus. This article elaborates on the research advances in persistent HBV infection from the aspect of virus, including covalently closed circular DNA, HBV particles, and HBV components.
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Objective To express HIV-1 capsid p24 antigen in an eukaryotic expression system and to evaluate its antigenicity and potential in the early diagnosis of HIV. Methods The full-length gene of HIV-1 p24 and the signal peptide DRVI gene were amplified by PCR. The signal peptide DRVI preceding the p24 gene was introduced using fusion PCR, and cloned into vector pDRVI1. 0. Two recombinant plas-mids pDRVI-p24 and pDRVI-p24s were constructed and transfected into 293F cells. Expression and secre-tion of p24 protein were detected by SDS-PAGE, Ni-NTA column chromatography and molecular sieve were used to purify p24s protein. The purified protein was identified by Western blot and indirect ELISA using hu-man/mouse HIV-1-positive serum samples. Results The eukaryotic expression system for HIV-1 p24 anti-gen was successfully established with high efficiency. The target protein of interest with the signal peptide DRVI was obviously detected in the supernatants of cell culture. The recombinant protein had good specifici-ty and sensitivity based on the results of serological tests using serum samples of five HIV-1-positive and five HIV-negative mice , 30 HIV-1-positive patients and 50 HIV-1-negative healthy individuals . Conclusions The eukaryotic expression system for HIV-1 p24 antigen was successfully established. The purified HIV-1 p24s antigen had good antigenicity. An indirect ELISA assay with good specificity and sensitivity for the de-tection of HIV-1 was preliminarily constructed and showed great potential for application.
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BACKGROUND: Transfusion-transmissible hepatitis B virus (HBV) infection is a major problem worldwide. Recently, confirmatory nucleic acid tests (NATs) for HBV DNA have been employed in several countries. We assessed the prevalence and yearly trends of HBV infection in blood donors in the Eastern Province of Saudi Arabia, screening for HBV surface antigen (HBsAg), antibody against HBV core antigen (anti-HBc), and HBV DNA. METHODS: Between 2011 and 2015, a total of 22,842 donors were screenedfor HBsAg, anti-HBc, and HBV DNA using the HBsAg Qualitative II kit (Abbott, Ireland Diagnostics Division, Sligo, Ireland), ARCHITECT Anti-hepatitis B core antigen antibody (HBc) II Assay kit (Abbott GmbH & Co. KG, Wiesbaden, Germany), and NAT Procleix Ultrio Elite Assay kit (Grifols Diagnostic Solutions Inc., Los Angeles, CA, USA), respectively. RESULTS: A total of 739 (3.24%) donors were HbsAg(+), anti-HBc(+), or HBV DNA(+); 63 (0.28%) were HbsAg(+), anti-HBc(+), and HBV DNA(+). Twelve (0.05%) were anti-HBc(+) and HBV DNA(+) but HBsAg(−); they were considered to have occult infection. Further, 664 (2.91%) were HBsAg(−) but anti-HBc(+), indicating chronic or resolving infection. HBV prevalence increased significantly from 2011 to 2012, increased marginally till 2013, and showed a decreasing trend from 2013 (P>0.05). CONCLUSIONS: The five-year prevalence of HBV infection among blood donors in the Eastern Province of Saudi Arabia (3.24%) is lower than that reported for other regions in the country. The occult HBV infection rate of 0.05% emphasizes the importance of NATs in isolating potential infectious blood units.
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Humans , Antigens, Surface , Blood Donors , DNA , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis B , Hepatitis , Ireland , Mass Screening , Prevalence , Retrospective Studies , Saudi Arabia , Tissue DonorsABSTRACT
Abstract INTRODUCTION: IgG subclasses involved in the immune response to hepatitis C virus (HCV) antigens have been rarely studied. We investigated the immune response mediated by IgG1 and IgG4 antibodies against the recombinant core and NS3 antigens in patients with chronic hepatitis C. METHODS: Sixty patients infected with HCV genotype 1 without antiviral treatment and 60 healthy subjects participated in the study. Serum levels of alanine aminotransferase, HCV viremia, and the presence of cryoglobulinemia and liver fibrosis were determined. We investigated the serum IgG1 and IgG4 antibodies against recombinant HCV core and NS3 non-structural protein antigens using amplified indirect ELISA. RESULTS: Anti-core and anti-NS3 IgG1 antibodies were detected in 33/60 (55%) and 46/60 (77%) patients, respectively, whereas only two healthy control samples reacted with an antigen (NS3). Anti-core IgG4 antibodies were not detected in either group, while 30/60 (50%) patients had anti-NS3 IgG4 antibodies. Even though there were higher levels of anti-NS3 IgG4 antibodies in patients with low viremia (< 8 × 105 IU/mL), IgG1 and IgG4 antibody levels did not correlate with ALT levels, the presence of cryoglobulinemia, or degree of hepatic fibrosis. High production of anti-core and anti-NS3 IgG1 antibodies was observed in chronic hepatitis C patients. In contrast, IgG4 antibodies seemed to only be produced against the NS3 non-structural antigen and appeared to be involved in viremia control. CONCLUSIONS: IgG1 antibodies against structural and non-structural antigens can be detected in chronic hepatitis C, while IgG4 antibodies seem to be selectively stimulated by non-structural HCV proteins, such as the NS3 antigen.
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Humans , Male , Female , Adult , Aged , Hepacivirus/immunology , Hepatitis C Antigens/immunology , Hepatitis C Antibodies/immunology , Hepatitis C, Chronic/immunology , Hepatitis C, Chronic/blood , Reference Values , Viremia , Immunoglobulin G/blood , Enzyme-Linked Immunosorbent Assay , Case-Control Studies , Statistics, Nonparametric , Hepatitis C Antigens/blood , Hepatitis C Antibodies/blood , Viral Load , Cryoglobulinemia , Alanine Transaminase/blood , Liver Cirrhosis/virology , Middle AgedABSTRACT
Occult hepatitis B infection (OBI) is a cause of concern while screening the blood donors to prevent transfusion-related transmission of infection. This study was conducted to assess the prevalence of OBI using total anti-HBc by ELISA and DNA detection by real time polymerase chain reaction (PCR). The samples included were negative for HBs Ag by ELISA. Out of 1102 samples tested, 156 were positive for total anti-hepatitis B core antigen and 52/156 by real-time PCR. Overall, the prevalence was found to be 4.71% (52/1102). The results indicate that nucleic acid-based testing should be an essential part of screening procedure to prevent missing of OBI.
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Objective@#To study the expression and purification of the chimeric virus-like particles displaying epitopes of EV71 as a candidate of enterovirus 71 gene recombined vaccinet.@*Methods@#The fusion protein hepatitis B core (HBc)-SP70 was constructed by inserting SP70 into the main immunogenic region of truncated hepatitis B core antigen (HBcAg) sequence, expressed in E. Coli, and purified through sonication, ion exchange chromatography, CsCl cushion centrifugation and density gradient centrifugation. Then its antigenicity was detected by ELISA and Western blot assay.@*Results@#Recombinant plasmid pHBc-SP70 was successfully constructed. And the soluble fusion protein was efficiently expressed induced by IPTG. The purity of the chimeric virus-like particles (VLPs) was up to 90% after the purification process described in method . The purified fusion protein HBc-SP70 could be spontaneously folded and assembled into empty virus-like particles and react with the monoclonal antibodies against HBc and SP70.@*Conclusions@#The chimeric VLPs displaying epitopes of EV71 were efficiently expressed and purified in E. Coli. with excellent antigenicity, which laid a foundation for evaluation of the immune effect evoked by this EV71 gene recombined vaccine.
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Objective@#To investigate the effect of hepatitis B core antigen (HBcAg) in promoting the invasion of hepatitis B virus (HBV)-related hepatocellular carcinoma cell line HepG2.2.15 and the role of Toll-like receptor 4 (TLR4) in the mechanism.@*Methods@#TLR4 mRNA and protein expression in HepG2 cells and HepG2.2.15 cells was measured by reverse transcription real-time PCR and Western blot analysis, respectively. HepG2.2.15 cells were transfected with TLR4 specific small interfering RNA (siRNA) to silence TLR4 expression, and stimulated by recombinant HBcAg in culture. The invasion of cells was measured by Transwell invasion assay. The expression of TLR4 signaling pathway-related proteins in the cultured cells and proinflammatory cytokines in the supernatant was also determined. The student’s t-test, one-way ANOVA, and SNK-q test were used for statistical analysis.@*Results@#TLR4 mRNA and protein expression in HepG2.2.15 cells was significantly higher than that in HepG2 cells. TLR4 siRNA transfection remarkably down-regulated TLR4 expression in HepG2.2.15 cells. Inhibiting TLR4 expression and/or HBcAg stimulation did not affect the proliferation of HepG2.2.15 cells. However, HBcAg stimulation significantly increased the invasion ability of HepG2.2.15 cells and the secretion of proinflammatory cytokines [including interferon (IFN)-γ, interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α]. Inhibiting TLR4 expression significantly reduced HBcAg-induced cellular invasion. Meanwhile, HBcAg stimulation elevated the expression of MyD88 and TRIF in HepG2.2.15 cells. TLR4 silencing inhibited HBcAg-induced increase in the expression of MyD88, while it showed no significant impact on TRIF expression.@*Conclusion@#HBcAg can promote the invasion of HepG2.2.15 cells. The TLR4/MyD88 signaling pathway may be involved in this process by inducing the expression of proinflammatory cytokines.
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Objective To evaluate the diagnostic value of hepatitis C virus core antigen(HCV-cAg),hepatitis C virus(HCV-IgG) and hepatitis C virus(HCV-RNA) in the laboratory diagnosis of Hepatitis C.Methods HCV-cAg and HCV-IgG were detected by enzyme-linked immunosorbent assay(ELISA),HCV-RNA was detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR) in 84 suspected HCV patients and 87 healthy control subjects.Results In 84 suspected HCV patients,the HCV-IgG positive rate was 84.5%,HCV-cAg positive rate was 13.1%,HCV-RNA positive rate was 52.4%.Among 71 cases of HCV-IgG positive patients,there were 35 cases with negative HCV-RNA,the false positive rate was 49.3%.In 11 cases of HCV-cAg positive patients,there were 5 cases with negative HCV-RNA,the false positive rate was 45.5%.In 44 cases of HCV-RNA positive diagnosis of hepatitis C patients,HCV-IgG false negative rate was 18.2%,HCV-cAg false negative rate was 86.4%.The false negative rate of combined detection of HCV-cAg and HCV-IgG was 13.6%,and the true positive rate was 100.0%.Conclusion HCV-cAg and HCV-IgG have certain false negative and false positive in laboratory diagnosis of HCV,combine these two methods,or joint with HCV-RNA detection,could reduce the rate of missed diagnosis.
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Objective To investigate the serum anti-HBc level in patients with different natural history of chronic hepatitis B virus (HBV) infection and cirrhosis,and its clinical value for distinguishing the natural history statue.Methods A total of 160 patients with chronic HBV infection from March 2015 to June 2016 were enrolled,and they were divided into immune tolerance group (n=43),HBeAg-positive chronic hepatitis B (CHB) group (n=37),inactive carrier group (n=39) and HBeAg-negative CHB group (n=41).A total of 44 patients with HBeAg-positive cirrhosis and 46 patients with HBeAg-negative cirrhosis were enrolled too.The general conditions data were collected,and HBsAg,HBeAg,anti-HBc,HBV DNA load and HBV genotype were detected.The associations between anti-HBc level and clinical parameters were analyzed,and the diagnostic value of anti-HBc for distinguishing different natural histories was analyzed.Results The anti-HBc levels in different natural history from high to low were as following: HBeAg-positive CHB group (4.22±0.68)log10 IU/mL,HBeAg-negative CHB group (3.89±0.88)log10 IU/mL,inactive carrier group (3.07±0.68)log10 IU/mL and immune tolerance group (2.88±0.82)log10 IU/mL.The anti-HBc levels in HBeAg-positive and HBeAg-negative cirrhosis patients were (3.04±0.82) and (3.15±0.86) log10 IU/mL,respectively.In HBeAg-positive CHB group,the anti-HBc was positively associated with ALT (r=0.353,P=0.032) and AST (r=0.421,P=0.009).In HBeAg-negative CHB group,the anti-HBc was positively associated with HBV DNA (r=0.343,P=0.028),ALT (r=0.458,P=0.003) and AST (r=0.495,P=0.001).The AUC of anti-HBc used to distinguish immune tolerance from HBeAg-positive CHB was 0.903,and the AUC used to distinguish inactive carrier from HBeAg-negative CHB was 0.833.Conclusion Anti-HBc levels in different natural history of chronic HBV infection are significantly different,and anti-HBc could be used to distinguish the natural history statue of chronic HBV infection with a higher diagnostic value than HBsAg.
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Objective To investigate the clinical significance of HCV core antigen (HCV-cAg )in hepatitis C screening. Methods From October 2014 to October 2015 ,a total of 8000 serum samples were collected from the outpatient and inpatient. HCV-cAg and hepatitis C antibody (HCV-Ab) were detected by using ELISA.The positive samples of HCV-cAg or HCV-Ab were performed the confirmative diagnostic detection by PCR.Results Among 8000 serum samples ,82 positive samples were screened out ,in which 73 cases were HCV-RNA positive by elementary screening with the standard of HCV-cAg or HCV-Ab positive ,a-mong which 72 were HCV-RNA positive and and 9 cases were HCV-RNA negative by HCV-RANA confirmation.The sensitivity and specificity of HCV-cAg were 45.83% and 99.98% ,sensitivity and specificity of HCV-Ab were 94.44% and 99.90% ,which of HCV-cAg and HCV-Ab combined detection were 100.00% and 98.86% respectively.Conclusion HCV-cAg and HCV-Ab play complementary role in HCV screening.Their combined detecting contributes to increase the screening rate of HCV.
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Objective To investigate the intensity of HBsAg and HBcAg expression in liver tissue of patients with chronic hepatitis B virus (HBV) infection and its clinical significance.Methods A total of 994 HBV infected patients underwent liver biopsy and histopathological examination.The expression of HBsAg and HBcAg in liver tissue was detected by histoimmunochemistry.Patients were divided into HBeAg (+)/HBVDNA(+), HBeAg(-)/HBV DNA(+) and HBeAg(-)/HBV DNA(-) groups according to HBeAg and HBV DNA levels;patients were divided into <2 × normal (ULN) group, 2-<5 × ULN groupand ≥5 × ULN group according to the alanine aminotransferase (ALT) levels.The histologic activity (A), fibrosis (F), the expression of HBsAg and HBcAg in liver tissue and their correlations with clinical features were analyzed.Logistic regression analysis was used to study the factors affecting the expression of HBsAg and HBcAg in liver tissue.Results Among 994 HBV infected patients, 941 cases (94.67%) were intrahepatic HBsAg positive and 553 cases (55.63%) were intrahepatic HBcAg positive;403 cases (40.85%) were ≥A2 in histologic activity and 371 cases (36.09%) were ≥F2 in fibrosis.The degree of A and F was the highest in HBeAg (-) / HBV DNA (+) group, followed by HBeAg (-) / HBV DNA (-) group, and was the lowest in HBeAg (+) / HBV DNA (+) group.The intensity of intrahepatic HBsAg expression was significantly different among three groups (x2 =6.299, r =-0.760, P < 0.05), however, the difference was not showed in pairwise comparisons.The difference of intrahepatic HBcAg intensity among three groups was statistically significant (x2 =282.995, r =-0.645, P < 0.01), the intensity was the highest in HBeAg (+) / HBV DNA (+) group and the lowest in HBeAg (-) / HBV DNA (-) group.The constituent ratio of HBeAg positive and HBV DNA level were higher and the average age was lower in intrahepatic HBsAg positive group than those in HBsAg negative group.The constituent ratio of positive HBeAg, the levels of ALT, AST, PLT and HBV DNA were higher and the average age, the average FIB-4 level were lower in intrahepatic HBcAg positive group than those in HBcAg negative group.The HBV DNA level was an independent risk factor for intrahepatic HBsAg intensity, and the HBeAg positive and HBV DNA level were independent risk factors for intrahepatic HBcAg intensity.There were no significant differences in A and F among different groups of intrahepatic HBsAg intensity (x2 =1.943 and 2.630, both P > 0.05).There was significant difference in F among different groups of intrahepatic HBcAg intensity (x2 =12.352, P < 0.01), but not in A.The degree of F was the highest in intrahepatic HBcAg negative group.There was significant difference in intrahepatic HBcAg intensity among different groups of ALT level (x2 =16.349, P < 0.01), but not in intrahepatic HBsAg intensity.The intrahepatic HBcAg intensity in ALT < 2 × ULN group was lower than that in other two groups.Conclusions Most of patients with chronic HBV infection are intrahepatic HBsAg positive and more than half of them are intrahepatic HBcAg positive.The intrahepatic HBsAg intensity is not associated with A and F, but correlates with HBV DNA level.The intrahepatic HBcAg intensity is not associated with A, but it is negatively correlated with F and positively correlated with positive HBeAg expression, HBV DNA level and ALT level.
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Objective:To prepare a virus-like particle (VLP),containing Hepatitis B virus core antigen (HBcAg) and N-terminal peptides of the L2 protein of human papilloma virus (HPV),and investigate the immunogenicity of the VLP in mice and the protection against different strains of HPV .Methods:A fusion gene was synthesized to insert a DNA fragment ,coding for the N-terminal epitopes of the L2 protein of HPV16,into the HBcAg coding sequence;HBc-L2 fusion protein was highly expressed in E.coli using the pET9a and BL21(DE3) expression system;the purified fusion protein was used to immunize BALB/c mice and antibody titers against the L2 epitopes in mouse sera were determined by indirect ELISA;the levels of neutralizing antibodies against both HPV 16 and 18 were also analyzed.Results:HBc-L2 fusion protein was expressed in E.coli and purified,with the purity >80%,by ammonium sulfate pre-cipitation and CL-4B gel filtration;analysis of the purified fusion protein ,using size exclusion chromatography with multi-angle laser light scattering detection ( SEC-MALS) and electron microscope ,revealed that HBc-L2 was assembled into a stable VLP structure auto-matically following its expression;immunization of BALB/c mice with the purified VLPs resulted in high antibody titers in mouse sera against the L2 epitopes;furthermore,it was demonstrated that the sera from the immunized mice had neutralization activities against both HPV16 and HPV18.Conclusion:The immunogenicity of the L2 epitopes was highly enhanced by the construction of HBc-L2 fusion protein and the formation of the VLP structure;the fusion protein was also capable of inducing protections against different serotypes of HPV,therefore,it could be a potential HPV vaccine with a broad coverage and low production cost .
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Hepatitis C virus core antigen (HCV Ag) is a recently developed marker of hepatitis C virus (HCV) infection. We investigated the clinical utility of the new HCV Ag assay for prediction of treatment response in HCV infection. We analyzed serum from 92 patients with HCV infection who had been treated with pegylated interferon and ribavirin. HCV Ag levels were determined at baseline in all enrolled patients and at week 4 in 15 patients. Baseline HCV Ag levels showed good correlations with HCV RNA (r = 0.79, P < 0.001). Mean HCV Ag levels at baseline were significantly lower in patients with a sustained virologic response (SVR) than in those with a non SVR (relapse plus non responder) based on HCV RNA analysis (2.8 log10fmol/L vs. 3.27 log10fmol/L, P = 0.023). Monitoring of the viral kinetics by determination of HCV RNA and HCV Ag levels resulted in similarly shaped curves. Patients with undetectable HCV Ag levels at week 4 had a 92.3% probability of achieving SVR based on HCV RNA assay results. The HCV Ag assay may be used as a supplement for predicting treatment response in HCV infection, but not as an alternative to the HCV RNA assay.
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Humans , Hepacivirus , Hepatitis C , Hepatitis C, Chronic , Hepatitis , Hepatitis, Chronic , Interferons , Kinetics , Ribavirin , RNAABSTRACT
Response-guided therapy is of limited use in developing countries because hepatitis C virus RNA detection by sensitive molecular methods is time- and labor-consuming and expen- sive. We evaluated early predictive efficacy of serum hepatitis C virus core antigen kinetics on sustained virologic response in patients with genotype 1 hepatitis C virus during pegylated interferon plus ribavirin treatment. For 478 patients recruited, hepatitis C virus RNAs were detected at baseline, and at weeks 4, 12, 24, 48, and 72 using Cobas TaqMan. Architect hepatitis C virus core antigen was performed at baseline, and weeks 4 and 12. Predictive values of hepatitis C virus core antigen on sustained virologic response were compared to hepatitis C virus RNA. In the first 12 weeks after treatment initiation the dynamic patterns of serum hepatitis C virus core antigen and hepatitis C virus RNA levels were similar in sustained virologic response, relapse, and null response patients groups. Although areas under the receiver operating characteristics curves of hepatitis C virus core antigen were lower than those of hepatitis C virus RNA at the same time points, modeling analysis showed that undetectable hepatitis C virus core antigen (rapid virological response based on hepatitis C virus core antigen) had similar positive predictive value on sustained virologic response to hepatitis C virus RNA at week 4 (90.4% vs 93.3%), and hepatitis C virus core antigen decrease greater than 1 log10 IU/mL (early virological response based on hepatitis C virus core antigen) had similar negative predictive value to hepatitis C virus RNA at week 12 (94.1% vs 95.Z%). Analysis on the validation group demonstrated a positive predictivevalue of 97.5% in rapid virological response based on hepatitis C virus core antigen and a negative predictive value of 100% in early virological response based on hepatitis C virus core antigen. In conclusion, hepatitis C virus core antigen is comparable to hepatitis C virus RNA in predicting sustained virologic response of chronic genotype 1 hepatitis C virus infected patients, and can be used to guide anti-hepatitis C virus treatment, especially in resource-limited areas.
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Adult , Female , Humans , Male , Middle Aged , Antiviral Agents/therapeutic use , Hepacivirus/immunology , Hepatitis C Antigens/immunology , Hepatitis C, Chronic/drug therapy , Interferon-alpha/therapeutic use , Polyethylene Glycols/therapeutic use , Ribavirin/therapeutic use , Genotype , Hepatitis C, Chronic/immunology , Hepatitis C, Chronic/virology , Predictive Value of Tests , ROC Curve , Recombinant Proteins/therapeutic use , Time Factors , Viral Core Proteins/immunologyABSTRACT
Objective To investigate the clinical characteristics,prevention and treatment strategy of de novo hepatitis B virus (HBV)infection after pediatric living liver transplantation.Methods In total,106 pediatric recipients undergoing living liver transplantation in Organ Transplantation Center of Affiliated Beijing Friendship Hospital of Capital Medical University and Organ Transplantation Center of Tianjin First Center Hospital from July 2010 to July 2014 were enrolled in this study.All surgeries were performed by the same surgical team.According to preoperative test outcomes of donor HBV serological markers,all recipients were divided into the positive (n =45)and negative (n =61)antibody to hepatitis B core antigen (anti-HBc)donor liver groups (positive group and negative group),and the prevalence of de novo HBV infection was compared between two groups.The risk factors of de novo HBV infection in the positive group were analyzed to elucidate the clinical characteristics of de novo HBV infection in affected children.Results The incidences of de novo HBV infection in positive and negative group were 18% (8 /45 )and 2% (1 /61 )respectively.The risk factors of de novo HBV infection in recipients with positive anti-HBc were negative anti-HBs before transplantation and absence of antiviral therapy post-transplantation in recipients (both in P <0.05 ).The median interval between time of onset and time of liver transplantation was 12 months (8-48 months).Seven cases were treated with lamivudine and the remaining two cases were left untreated.All nine recipients survived.Conclusions Application of positive anti-HBc donor liver have a risk of HBV infection in recipients after pediatric liver transplantation.Absence of postoperative nucleoside analogue therapy and negative anti-HBs before transplantation acts as risk factors of de novo HBV infection in the recipients with positive anti-HBc donor liver.After liver transplantation,nucleoside analogue therapy is recommended for the pediatric recipients with positive anti-HBc donor liver to prevent the incidence of de novo HBV infection.Besides,hepatitis B vaccine should be administered prior to liver transplantation.