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Objective:To establish the hepatic organoid of hepatitis B virus (HBV) infection on the basis of induced pluripotent stem cells (iPSC) and an inverted colloidal crystal polyethylene glycol scaffold (ICC), and to evaluate the antiviral effect of nucleoside drugs.Methods:iPSC was differentiated into hepatocyte-like cells (HLC), and inoculated into ICC to construct a hepatic organoid. The relative mRNA expressions of Nanog homeobox (NANOG), sex determining region Y-box (SOX) 2, SOX17, forkhead box protein A2 (FOXA2), alpha fetoprotein (AFP) and albumin (ALB) were detected by real time quantitative polymerase chain reaction (RT-qPCR). Confocal laser microscopy was used to photograph the three-dimension (3D) structure of organs. The expression of sodium taurocholate cotransporting polypeptide (NTCP) in HLC was analyzed by Western blot and immunofluorescence. HepG2.2.15 cells were used to extract HBV virus particles to infect hepatic organoid. The relative expression of HBV pregenome RNA (pgRNA) in cells was detected by RT-qPCR. The expressions of hepatitis B core antigen (HBcAg) and hepatitis B surface antigen (HBsAg) in cytoplasm were observed under confocal laser microscopy. A total of 0.5 μmol/L entecavir and 0.5 μmol/L lamivudine were used to treat the infected cells respectively. The relative expression of HBV pgRNA in infected and uninfected cells was detected by RT-qPCR. Independent sample t test and one-way analysis of variance were used for statistical analysis. Results:Within 21 days of iPSC differentiation, the mRNA expressions of NANOG and SOX2 in stem cells markers decreased ( F=158.90 and 8.31, respectivley; P<0.001 and P=0.002, respectively), while the mRNA expressions of SOX17 and FOXA2 in the endoderm increased first and then decreased ( F=37.23 and 82.57, respectively, both P<0.001). In the later stage of differentiation, the mRNA expressions of AFP and ALB in liver cells increased ( F=4.65 and 34.64, respectively, P=0.012 and P<0.001, respectively), and all differences were statistically significant. NTCP was highly expressed in differentiated cells detected by Western blot and fluorescence microscopy, the protein expression level was 0.803±0.099. Confocal laser microscopy confirmed that the differentiated cells expressed ALB and presented spherical structure in ICC. The expression of HBV pgRNA and the immunostaining of HBsAg and HBcAg confirmed that HBV successfully infected hepatic organoid. Three days after the application of entecavir and lamivudine, the HBV pgRNA level decreased significantly both in entecavir group (0.665±0.220) and lamivudine group (0.503±0.117) compared to the uninfected cells (3.347±0.454), and the differences were both statistically significant ( t=10.53 and 12.72, respectively, both P<0.001). Conclusions:HLC display hepatic specific genes ALB and NTCP. Hepatic organoids constructed with iPSC and ICC have human liver function and can be infected by HBV. Entecavir and lamivudine could effectively inhibit the replication of HBV in hepatic organoids.
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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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ObjectiveTo clarify the evolutionary laws of syndromes and syndrome elements at different stages during the malignant transformation of chronic hepatitis B （CHB）. MethodsA total of 671 patients with hepatitis B virus infection， who were admitted to the outpatient and inpatient departments of Dongzhimen Hospital of Beijing University of Chinese Medicine and The Fifth Medical Center of Chinese PLA General Hospital from July 1st， 2020 to June 30th， 2021， were included， involving 120 cases of CHB， 340 cases of hepatitis B liver cirrhosis （HBLC）， 64 cases of precancerous lesions with hepatitis B liver cirrhosis （PLHC）， and 147 cases of hepatitis B liver cirrhosis with hepatocellular carcinoma （HCC）. A Survey form of traditional Chinese medicine syndrome during malignant transformation of chronic hepatitis B was designed， and the general information， auxiliary examination and the four examinations results were collected. Factor analysis and K-means clustering were used to determine and statistically analyze the syndrome and syndrome elements. ResultsFive traditional Chinese medicine （TCM） syndrome types were identified in CHB patients， while there were six TCM syndrome types in HBLC， PLHC and HCC stages. Among CHB patients， the main syndromes were liver constraint and spleen deficiency （53.33%） and liver-gallbladder damp-heat （21.67%）， and the dominant syndrome elements were qi stagnation （27.60%）， heat （17.71%） and qi deficiency （17.71%）. In the HBLC stage， the syndromes were mainly blood stasis obstructing the collaterals （23.83%） and liver constraint and spleen deficiency （22.35%）， with dominant syndrome elements being blood stasis （19.25%）， dampness （17.46%）， and qi deficiency （15.01%）. For the PLHC stage， the primary syndrome types were blood stasis obstructing the collaterals （29.68%） and liver-kidney yin deficiency （20.31%）， and the leading syndrome elements were blood stasis （22.12%）， yin deficiency （15.93%）， and qi deficiency （15.04%）. In the HCC stage， the syndrome was dominated by blood stasis obstructing the collaterals （33.34%） and liver-kidney yin deficiency （19.73%）， with the main syndrome elements being blood stasis （24.52%）， yin deficiency （16.09%）， and qi deficiency （15.33%）. During the progression of CHB to malignancy， there was a gradual decrease in excess syndromes including liver-gallbladder damp-heat and water-dampness internal obstruction from 21.67% to 19.04%. In contrast， deficiency syndromes including liver-kidney yin deficiency and spleen-kidney yang deficiency increased from 15.83% to 31.97%. Additionally， excess syndrome elements including qi stagnation， heat and dampness decreased from 59.89% to 34.48%， while deficiency syndrome elements including qi deficiency， yin deficiency and yang deficiency increased from 32.30% to 41.00%. ConclusionDuring the malignant transformation of CHB， there exists a progression of syndrome and syndrome elements， shifting from qi stagnation， heat and qi deficiency to blood stasis （predominantly excess）， dampness and qi deficiency， and then to blood stasis （predominantly deficiency）， yin deficiency and qi deficiency， characterized by “deficiency-excess complex， and shift from excess to deficiency”.
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【Objective】 To analyze the characteristics of gene mutation in S region of blood donors with occult hepatitis B virus infection (OBI) in Huzhou area. 【Methods】 A total of 60 107 blood samples in Huzhou between October 2018 and June 2020 were collected by our blood station. Among them, 52 samples were NAT, yield and their epidemiological characteristics were analyzed. Twenty-seven OBI out of the 52 NAT yield samples were included in experimental group. Other eight HBV-infected individuals with positive HBsAg, core antibody (anti-HBc) and HBV-DNA were selected as positive control. Liver function and 5 serological markers of HBV were compared between the two groups, and HBV genotypes and amino acid mutation in S region in the two groups were analyzed. 【Results】 The number of NAT-yield samples were different by gender, age, and educational background (P0.05). Surface antigen (HBsAg) in the experimental group was significantly lower than that in the control group, while surface antibody (anti-HBs) and e antibody (anti-HBe) were significantly higher than those in the control group (P<0.05). Twenty sequences in S region were obtained from the experimental group, including 4 in S region and 16 in preSS region; 16 cases with type C and 4 cases with type B. 【Conclusion】 The follow-up of NAT-yield blood donors in Huzhou area should be conducted. Compared with HBV infected individuals with positive HBsAg, anti-HBc and HBV-DNA, those with OBI have a higher gene mutation rate in S region.
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【Objective】 To investigate the prognosis of blood donors with occult hepatitis B virus infection (OBI) by long-term follow-up and repeated testing of HBsAg and HBV DNA. 【Methods】 From January 1, 2010 to December 31, 2020, voluntary blood donors were screened by both serological and viral nucleic acid(NAT) testing, then samples were further confirmed as HBV DNA positive by manual nested-PCR amplification.A total of 306 cases were detected as HBsAg negative /HBV DNA positive, then followed-up for a long time and re-examined of HBsAg and HBV DNA to confirm whether they had infected with OBI.The prognosis of patients with OBI who experienced long-term immunization was determined by repeated testing. 【Results】 A total of 306 HBsAg negative/ HBV DNA positive blood donors had been followed up, and 40(13.07%, 40/306) were recalled frequently for re-examination.Among them, 90%(36/40), 57.5%(23/40), 40% (16/40)were anti-HBc + , anti-HBs + and anti-HBe + , respectively, and 50%(20/40), 40%(16/40), 7.5%(3/40) and 2.5% (1/40)were anti-HBs+ / anti-HBc + , anti-HBc + / anti-HBs -, anti-HBc -/ anti-HBs + and anti-HBc -/ anti-HBs -, respectively.Those 40 blood donors were followed-up for 1-13 times, with the duration of 8-108 months (0.6~9 years).1 donor (2.5%) was followed-up less than 1 year, 11 (27.5%)>1 year and ≤3 years, 23 (57.5%) 23(57.5%)>3 years and ≤5 years, and 5 (12.5%) for more than 5 years.After long-term following up and repeated testing, 50%(20/40)of OBI blood donors turned negative for HBV DNA (HBsAg negative / HBV DNA negative), 42.5% (17/40)were confirmed as OBI infection (HBsAg negative / HBV DNA positive), and 7.5%(3/40) were hard to determine (after repeated testing, the results were either positive or negative). 【Conclusion】 After long-term following up and repeated screening, we found that none of the OBI patients turned into acute or chronic HBV infection, and most of them maintained OBI.However, OBI blood donors carry very low load of HBV DNA for a long time, which could lead to false negative results of NAT and bring a great challenge to the safety of blood transfusion.
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Background and Aim: Globally, hepatitis B virus (HBV) infection is among the commonest chronic infections and the leading cause of liver cancer. This study evaluated inflammatory and liver injury biomarkers among newlydiagnosed HBV-infected patients to reveal inflammation and liver injury levels. Patients and Methods: This case-control study was conducted among 146 newly diagnosed drug-naive patients and 64 blood donors. Questionnaires were administered to obtain demographic data. Blood samples were collected to assess viral serological markers, inflammatory markers, liver function, and hematological indices. Also, noninvasive markers of liver fibrosis (APRI: aspartate transaminase - platelet ratio index, FIB-4: fibrosis 4 index, and AAR: aspartate - alanine transaminase ratio) were mathematically derived. The patients were categorized into acute and chronic infections based on their viral serological markers. Results: Overall, 81.5% of the patients had an acute HBV infection, whereas 18.5% had a chronic HBV infection. There was a significant increase in the biomarkers of inflammation, C-reactive protein (CRP) and interleukin 6, and liver injury (liver transaminases, FIB-4 index, and APRI) among the drug-naive chronic HBV-infected patients. The study also revealed significant anemia and leucocytosis in patients with chronic HBV infection. Further, the study showed a strong correlation between CRP and alanine transaminase among patients with chronic HBV infection. Conclusion: There was increased anemia, inflammation, and liver fibrosis among the drug-naive chronic HBVinfected patients; hence, public education is required so patients with viral hepatitis B in Ghana would visit the clinic earlier enough for proper clinical management.
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Hepatitis B virusABSTRACT
Objective To investigate the immune regulation and prognosis of hepatitis B virus (HBV) infection in infected patients, and to analyze the relationship between immune regulation and clinical outcome. Methods The data of 633 cases of HBV infection in Huhhot from January 2015 to December 2018 were retrospectively analyzed. At the same time, healthy people from the Affiliated Hospital of Inner Mongolia Medical University were selected as the control group. Immune cell regulatory factors and lymphocyte subsets in blood of HBV infected and healthy people were tested. The results of one-year clinical outcomes were calculated. The relationship between immune cell regulatory factors and lymphocytes and the clinical outcomes of patients was analyzed by multiple logistics regression. Results The levels of CD4+T cells, CD8+T cells, NK cells, IL-2, and IL-12 in the blood of patients with HBV infection were significantly lower than those in the control group, and decreased as the patients’ clinical outcome deteriorated (P0.05). Low CD4+ T cells, low CD8+ T cells, low NK cells, low IL-2, low IL-12, high IL- 4 and high IL-10 were independent risk factors leading to the deterioration of clinical outcomes (P<0.05). Conclusion The immune regulation of patients with HBV infection is in a state of suppression, which is an independent factor affecting the clinical outcome of patients.
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Objective To explore the impact of maternal hepatitis B (HBV) virus infection status on the vaccination effect of the offspring. Methods Eighty-six pairs of HBV-infected mothers and newborns who were treated in Dalian Sixth People's Hospital from 2017 to 2019 were selected as the research subjects. According to the strength of the hepatitis B vaccine response, all patients were divided into two groups, the observation group (none or weak response) and the control group (strong response). Hepatitis B virus infection was compared between the mothers of the two groups, and multivariate logistic regression was used to analyze the influencing factors of vaccination effect of the offspring. Results The positive rate of HBV DNA in mothers of the observation group was higher than that in the control group. HBV DNA was mainly at high load and HBeAg was at low level. The HBV DNA of the control group was mainly at low load, while the distribution of HBeAg was close. The difference was statistically significant (P "HBsAg(+), HBeAg(+), anti-HBc(+)" in 14 cases (51.85%), "HBsAg(+), anti-HBe(+), anti- HBc(+)" in 11 cases (40.74%) , while there were 27 cases (45.76%), 25 cases (42.37%), and 7 cases (11.86%), respectively, in the control group. The difference was statistically significant (P<0.05). Multivariate logisitic regression analysis showed that HBV DNA positivity, HBV DNA high load, HBeAg positivity, and low HBeAg levels were risk factors for weak response of the offspring vaccination. Conclusion HBV DNA positivity, high HBV DNA load, HBeAg positivity, and low HBeAg levels in HBV-infected mothers were risk factors for weak response of the offspring to vaccines, which should be paid attention to in clinic.
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Humans , Infant , Hepatitis B , Hepatitis B, Chronic , Hepatitis , Hepatocytes , Immunization , Immunization Programs , Immunotherapy , Prevalence , Vaccination , World Health OrganizationABSTRACT
Background@#Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.@*Methods@#The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.@*Results@#Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ2 = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response.@*Conclusion@#Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission.@*Trial Registration@#NCT01962155; https://clinicaltrials.gov.
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Objective To investigate the clinical features of hepatitis B core antibody (anti-HBc)positive patients with liver injury.Methods A total of 212 anti-HBc positive and HBsAg negative patients who were primarily diagnosed with liver injury from August 2013 to August 2014 at Ruijin Hospital were collected for this study.The patients were divided into cirrhosis group (n=60) and non-cirrhosis group (n =152) according to the status of cirrhosis.The 60 cases with cirrhosis were further compared with 60 cases with post-hepatitis B cirrhosis.The general information,biochemistry and immunology data were assessed.ANOVA was used to compare multiple groups of means,and Wilcoxon rank-sum test was used for non-parametric comparisons of the two groups.Results Only one case was positive for HBV DNA with the positivity rate of 0.5％.The causes for liver injury were as follows,60 cases with cryptogenic cirrhosis,which accounted for 28.3 ％;45 cases with drug-induced hepatitis,which accounted for 21.2 ％;33 cases with unexplained liver injury,which accounted for 15.6％;28 cases with acute hepatitis E,which accounted for 13.2％ and 15 cases with autoimmune hepatitis,which accounted for 7.1％.There were significant differences of T cell subpopulation,hepatitis B surface antibody (HBsAg) and hepatitis B e antibody (anti-HBe) quantitative level,red blood cells (RBC),platelet counts (PLT),prealbumin,albumin,alamine aminotransferase (ALT),aspartate transaminase (AST),international normalized ratio (INR),hyaluronic acid (HA),collagen Ⅲ (COL-Ⅲ) and collagen Ⅳ (COL-Ⅳ) between the cirrhosis group and non-cirrhosis group (all P＜0.05).The CD3+ CD4+ and CD3+ CD8+ counts,white blood cells (WBC),ALT,AST,total bilirubin (TBil) and albumin in anti-HBc-positive cirrhosis group were statistically different from those in post-hepatitis B cirrhosis group (all P＜0.05).Conclusions Some patients with positive anti-HBc still have HBV replication and infectivity.HBV anti-HBc positivity and HBsAg negativity may be associated with some cryptogenic cirrhosis and primary liver cancer.Patients with positive anti-HBc are prone to be complicated with drug-induced hepatitis,autoimmune hepatitis,and other liver damage related to immune mechanisms.Patients with cirrhosis have a higher risk to induce immune tolerance and progress to chronic disease than non-cirrhotic patients.Quantitative anti-HBc might be used as an indicator to predict disease progression after HBV infection.Disease condition in cirrhotic group with positive anti-HBc and negative HBsAg is less severe than that in post-hepatitis B cirrhosis group.
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BACKGROUND/AIMS: The objective of our study was to determine the epidemiological, laboratory, and serological characteristics of patients with chronic hepatitis B virus (HBV) infection and normal transaminases. The study also aimed to evaluate liver damage by measuring the liver fibrosis (LF) grade and to identify possible factors associated with the presence of fibrosis. METHODS: A retrospective observational study was conducted in patients with chronic HBV infection and classified as inactive carriers or immune-tolerant. Epidemiological variables of age, sex, immigrant, alcohol consumption, and body mass index (BMI), as well as virological variables (HBV DNA) and transaminase level were collected throughout the follow-up. The LF grade was evaluated by transient elastography. The cutoff value for significant fibrosis (SF) was liver stiffness ≥7.9 kPa. RESULTS: A total of 214 patients were included in the analysis, and 62% of them had a BMI ≥25 kg/m². During follow-up, 4% of patients showed transaminase elevation ( < 1.5 times normal). Most patients had a viral DNA level < 2,000 IU/mL (83%). Data on LF were available in 160 patients; of these, 14% had SF, 9% F3, and 6% F4. The variables associated with the presence of SF were transaminase alteration during follow-up, as 23% of patients with SF had elevated transaminases versus 3% of patients without SF (P < 0.005), and BMI, as the vast majority of patients with SF (88%) had a BMI ≥25 kg/m² versus 56% of patients without SF (P < 0.05). CONCLUSIONS: In patients with chronic HBV infection and normal transaminases, liver damage does not seem to be related to DNA levels, alcohol consumption, or immigrant status. SF seems to be associated with transaminase alteration during follow-up and elevated BMI. It is therefore recommended to measure LF grade with validated non-invasive methods in such patients.
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Humans , Alcohol Drinking , Body Mass Index , DNA , DNA, Viral , Elasticity Imaging Techniques , Emigrants and Immigrants , Fibrosis , Follow-Up Studies , Hepatitis B e Antigens , Hepatitis B, Chronic , Hepatitis, Chronic , Liver Cirrhosis , Liver , Observational Study , Retrospective Studies , TransaminasesABSTRACT
Objective@#To analyze the residual risk of transfusion transmitted hepatitis B virus (HBV) infection by enzyme-linked immunosorbent assay (ELISA) method in hepatitis B surface antigen (HBsAg) negative blood donors, and to assess the infection status.@*Methods@#A total of 45551 samples were collected from blood donors.All samples were tested by 2 different ELISA kids of HBsAg and nucleic acid testing (NAT) individually of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV). Those ELISA HBsAg negative and NAT single reactive (HBsAg-/HBV DNA+ ) specimens were analyzed by quantitative detection of HBV DNA and by serologic testing of HBV antigen and antibody.@*Results@#A total of 44 HBsAg-/HBV DNA+ samples were detected, including 42 occult HBV infections (OBI) and 2 window period infections (WP). The detection rate of OBI rate was 0.90‰, and 32 samples of OBI sample HBV DNA was less than 20 IU/ml, and the OBI detection rate was significantly different between different genders, ages and blood donation times (P<0.05). In the OBI sample, there were 6 serological models, 92.9%(39/42) OBI samples hepatitis B core antibody (HBcAb) positive, and 76.9%(30/39) HBV DNA in HBcAb positive samples were less than 20 IU/ml; 29.5% (13/42) of OBI blood donors hepatitis B e antigen (HBeAb) and HBcAb were also positive, of whom 84.6% (11/13) were HBV DNA quantitatively <20 IU/ml.@*Conclusions@#HBV residual risk of transfusion-transmitted infection may occur through HBsAg- and single NAT reactive blood donors, mainly include OBI, and HBV DNA low level. Blocking of single NAT reactive blood donors could reduce transfusion-transmitted HBV infection.
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The level of viral DNA in patients with occult hepatitis B virus infection (OBI) is very low, and it is difficult to detect the conventional serum marker-HBsAg. OBI brought challenges to the clinical diagnosis and treatment and blood transfusion safety. The mechanism involved in OBI and the clinical implication are getting more and more attention. OBI has a complex mechanism that may involve host factors, the virus itself, and other viral or nonviral factors. OBI has the risk of HBV transmission. HBV can be reactivated and the liver disease can be aggravated in patients with OBI.
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Hepatitis associated anti-tuberculous treatment (HATT) has been a main obstacle in managing patients co-infected with Mycobacterium tuberculosis and hepatitis B virus (HBV).Therefore,we evaluated the factors related to the severity of adverse effects during HATT,especially those associated with liver failure.A retrospective study was carried out at Tongji Hospital from 2007 to 2012.Increases in serum transaminase levels of＞3,5,and 10 times the upper limit of normal (ULN) were used to define liver damage as mild,moderate,and severe,respectively.Patients with elevated total bilirubin (TBil) levels that were more than 10 times the ULN (＞171 μrnol/L) with or without decreased (＜40％) prothrombin activity (PTA) were diagnosed with liver failure.A cohort of 87 patients was analyzed.The incidence of liver damage and liver failure was 59.8％ (n=52) and 25.3％ (n=22),respectively.The following variables were correlated with the severity of hepatotoxicity:albumin (ALB) levels,PTA,platelet counts (PLT),and the use of antiretroviral therapies (P＜0.05).Hypo-proteinemia and antiretroviral therapy were significantly associated with liver failure,and high viral loads were a significant risk factor with an odds ratio (OR) of 2.066.Judicious follow-up of clinical conditions,liver function tests,and coagulation function,especially in patients with high HBV loads and hypoalbuminemia is recommended.It may be advisable to reconsider the use of antiviral drugs failure during the course of anti-tuberculous treatment of HBV infection patients to avoid the occurrence of furious liver failure.
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Objective To explore the biological and serological characteristics of occult HBV infection among qualified young blood donors with anti-HBc positive in Suzhou area .Methods 120 young blood donors with negative HBsAg and reactivity in NAT in our hospital from October 2013 to June 2016 were selected as the research subjects .The anti-HBs quantitative detection and two pairs of semi-hepatitis B detection were performed .The samples of anti-HBc positive were conducted the viral nucleic acid extraction and nested PCR at the basic core promoter(BCP) region ,pre-core(PC) region and S region .The samples with positive amplification results were performed the gene sequencing and sequential analysis .Results Among 120 volunteer blood donors ,anti-HBc(+ ) had 31 cases ,in which 25 cases were in the 22-25 years old group and 6 cases in the 18-21 years old group ,the difference was statisti-cally significant(P<0 .05);anti-HBs(+ ) had 89 cases ,among them ,16 cases were in the 22 -25 years old group and 73 cases in the 18-21 years old group .The nest PCR was used to conduct PCR amplification in 31 samples of anti-HBc(+ ) ,in which 1 case was BCP region positive and 2 cases were S region positive ,all belonged to the 22-25 years old group .In the typing and sequencing for the samples of S region positive ,the results showed that all 2 cases were B type HBV .Compared with wild type DNA sequence , among them 1 cases had amino acid sequence E44U variation and 1 cases had T532G mutation .Conclusion For the anti-HBc(+ ) young blood donors qualified in HBsAg detection ,it is not guaranteed that their blood do not contain HBV DNA .It is still needed to further increase the sensitivity of nucleic acid detection method in hepatitis B highly prevalent areas fro decreasing the risk of HBV transmission by blood transfusion .
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Objective To study the prevalence of the occult hepatitis B virus infection (OBI) and the mutation of amino acid sequence in S gene of voluntary blood donors in AnHui/FuJian/Jiang Xi Province Blood centers.Methods Serologic testing for anti-HBc by ELISA was performed with HBsAg-HBV DNA+ samples from voluntary blood donors in three province blood centers.The S region of HBV of those samples was amplified and sequenced.The genotype and mutation of amino acid sequence were analyzed by MEGA6.Results 21 in 123046 blood donors from AnHui Province blood center were HBsAgHBV DNA+,the prevalence of OBI was 0.017％,and 76.2％ of these-OBI samples was positive in anti-HBc,S region was amplified by nest-PCR in 15 OBI samples,8 of them were B genotype,the others were C genotype.39 samples of 51 OBI blood donors from FuJian Province blood center were anti-HBc positive,16 samples of those OBI donors were amplified S region,14 were B genotype,the others were C genotype.There are 30 OBI blood donors from JiangXi Province blood center,24 of them were anti-HBc positive,S region was amplified in 4 samples,1 was B genotype,the others were C genotype.Of all 35 OBI samples,26 showed amino acid mutation,which was in MHR region of S gene,especially in HBV α epitope.Conclusion The rate of prevalence of OBI in AnHui Province was 0.017％,there was also certain OBI infection in FuJian and JiangXi Province.In the OBI samples which were amplified S region,the positive rates of anti-HBc in three blood centers were 73.3％,93.8％,100％.B Genotype was the main HBV genotype.The mutation in MHR region of S gene,especially in HBV α epitope,may be one of the reasons to cause OBI.
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Objective To investigate the interrelated liver damage and hepatitis B virus infection among breast cancer patients after chemotherapy, to provide guidance for future breast reduction combined hepatitis B virus infection after chemotherapy liver damage.Methods120 cases of breast cancer patients undergoing chemotherapy combined hepatitis B carries from June 2012 to November 2016 in ningbo women and children's hospital were selected as the research object, depending on whether the infection with the hepatitis B virus into the study group and the control group, the study group HBV-DNA, HBsAg are positive, totaling 62 cases;control group, HBV-DNA, HBsAg were negative, totaling 58 cases;compare two groups of patients after chemotherapy in cases of liver damage.ResultsThe study group after chemotherapy, the incidence of liver dysfunction 48.28% in the control group after chemotherapy, the incidence of liver dysfunction 6.45 percent, the study group after chemotherapy, the incidence of liver dysfunction was significantly lower than the control group, the difference was statistically significant (P<0.05).Study group Ⅰ liver damage degree, degree Ⅱ, degree Ⅲ, degree Ⅳ of apparent higher, the difference was statistically significant (P<0.05), antiviral therapy 20 cases, no antiviral treatment in 42 cases.Antiviral therapy HBV reactivation rate and incidence of liver dysfunction were 5.0%, 20.0%;no antiretroviral therapy in HBV reactivation rate and the incidence of liver dysfunction 31.0%, 52.4% respectively;HBV antiviral therapy re-activation rate and the occurrence of liver dysfunction were significantly lower than not antiviral therapy, and the data were statistically significant (P<0.05).ConclusionThe clinical having close links between liver damage and breast cancer combined hepatitis B virus infection with hepatitis B virus are more likely to occur after infection liver dysfunction chemotherapy, and breast cancer patients after chemotherapy.
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Objective To explore the risk factom of hepatitis B virus infection and alcoholic liver disease in patients with decompensated cirrhosis complicated with liver failure.Methods One hundred and fifty-eight cases hepatitis B virus infection and alcoholic liver disease in patients with decompensated cirrhosis were selected.According to whether complicated with liver failure,the patients were divided into observation group with 62 cases (complicated with liver failure group) and control group with 96 cases (without liver failure group).The clinical data and results of 2 groups were analyzed to screen the risk factors of liver failure.Results Compared with control group,observation group in alanine aminotransferase,aspartate aminotransferase,cholinesterase,total bilirubin,and prothrombin time,activated partial thrombin time live enzymes,thrombin time,fibrinogen,serum creatinine,the differences were not statistically significant (P ＞ 0.05);in albumin ((28.02±7.36) g/L vs.(23.26±6.54) g/L,t =4.421,P =0.002),serum urea nitrogen ((8.84±4.71) mmol/L vs.(9.33±5.24) mmol/L,t =3.656,P=0.007),upper gastrointestinal bleeding(x2 =7.534,P=0.006),ascites (x2 =8.615,P =0.003),infection (x2 =10.321,P =0.001),hepatic encephalopathy (x2 =6.561,P =0.010),hepatorenal syndrome(x2 =4.952,P=0.026),the difference were statistically significant.(2)The results of logistic regression analysis showed that upper gastrointestinal bleeding (OR =1.020,95％ CI:1.003-1.036),hepatorenal syndrome(OR=2.872,95％CI:0.385-21.423) were risk factor of hepatitis B virus infection and alcoholic liver disease in patients with decompensated cirrhosis complicated with liver failure.Conclusion Upper gastrointestinal bleeding,hepatorenal syndrome are independent risk factors of hepatitis B virus infection and alcoholic liver disease in patients with decompensated cirrhosis complicated with liver failure.
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Objective To analyze the mutation of PreS-S region in occult hepatitis B virus(OHBV) in HBV infected persons with positive HBsAb and investigate the biological mechanisms of the special infectious model.Methods A total of 38 HB-sAb positive OBI serum samples were amplified by Nested PCR and sequenced,HBV genotype and serotype were determined.The amino acid sequences of OHBV were compared to the corresponding sequence of wild-type strains of similar genotype obtained from the GenBank database.Results PreS-S segment of 11 samples were obtained and 8 samples were sequenced successfully.Among which,5 were genotype C and 3 were genotype B.Genotype B were all serotype adw,while genotype C were 1 adw and 4 adr.The mutation rates of PreS-S region,the immunoreactive area and the major hydrophilic region (MHR) were higher in OHBV than the wild-type strains (2.6％ vs 0.8％,x2 =40.23,3.2％ vs 0.3％,x2 =52.13,3.6％ vs 0.6％,x2 =13.25,all P＜0.01) and the substitutions of I126T,Q129R,M133T,F134I,D144E,G145K in α determinant were found in OBI samples.The mutation rate of amino acids in PreS-S region was higher in genotype C than genotype B (3.5％ vs 1.2％,x2--15.98,P＜0.01),meanwhile,the mutation rates in MHR,α determinant and immunoreactive region were higher in genotype C too,but no statistical significance was attained (4.7％ vs 1.7 ％,x2 =2.96,3.6 ％ vs 2.9％,x2 =0.25,4.1％ vs 2.3％,x2 =3.59,all P ＞0.05).Conclusion Mutations in PreS-S region,especially in immunoepitope,might change the virus'immunogenicity leading to escape from immune response and cause OBI with HBsAb positive.