Objective To explore the potential mechanism of severe liver injury shortly after withdrawal of antiviral therapy in chronic hepatitis B (CHB) patients.Methods Forty-nine patients with chronic hepatitis B virus (HBV) infection from the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University and 8 healthy volunteers from August 2014 to March 2015 were included in this study.All of them were human leukocyte antigen (HLA)-A2-positive.CHB patients were classified into three groups,including 15 cases in immune-tolerance group,20 cases in sustained antiviral treatment group,and 14 cases in recurrence of drug withdrawal group.The frequency of peripheral HLA-A0201-restricted hepatitis B core antigen (HBcAg)18-27 pentamer complex specific CD8+ T cells in CHB patients was analyzed by flow cytometry.Enzyme linked immunospot assay(ELISPOT) was used to detect interferon-gamma (IFN-γ) and tumor necrosis factor-α (TNF-α) secretions of HBcAg18-27-specific CD8+ T cells.The experimental data were analyzed using non-parametric U tests.Results In healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group,the frequencies of HBcAg-specific CD8+T cells were (0.17 ± 0.16) ％,(1.46±0.72)％,(3.24± 1.60)％ and (4.67±2.43)％,respectively.Compared with healthy control group,the difference were all statistically significant in the three groups (Z=-3.583,-4.018 and-3.823,respectively;all P＜0.01).The frequencies of HBcAg-specific CD8+T cells in immune tolerance group or recurrence of drug withdrawal group were both significantly different from that in sustained antiviral therapy group (Z=-3.400 and-2.030,respectively;both P＜0.05).The difference between immune-tolerance group and recurrence of drug withdrawal group was also significant (Z =-3.230,P＜0.01).The secretion levels of IFN-γ of HBcAg-specific CD8+T cells in healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group were2 (0-6),16 (2-53),106 (14-254) and 156 (28-395) spot forming cell (SFC)/106 peripheral blood mononuclear cell (PBMC),respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treatment group or recurrence of drug withdrawal group were all statistically significant (Z=-3.585,-4.069 and-3.824,respectively;all P＜0.01).The IFN-γ level of HBcAg-specific CD8+ T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.205,P=0.027),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.700,P＜ 0.01).The TNF-α levels secreted by HBcAg-specific CD8+ T cells in each group were 2 (0-5),16 (2-32),112 (15-283),and 195 (55-537) SFC/106PBMC,respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treament group or recurrence of drug withdrawal group were all statistically significant (Z=-3.619,-4.069 and-3.824,respectively;all P＜0.01).The TNF-α level secreted by HBcAg-specific CD8+T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.449,P=0.014),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.350,P＜0.01).Conclusions The changes of frequency and immune function of HBcAg-specific CD8+T cells in CHB patients may be one of the reasons causing severe liver damage after irregular withdrawal of nucleoside analogues.

ObjectiveTo analyze the levels of matrix metalloproteinase 9 (MMP-9 ) in cerebrospinal fluid (CSF) of patients with central nervous system (CNS) infection caused by different pathogens.MethodsThe levels of MMP 9 in CSF were detected by enzyme linked immunosorbent assay (ELISA) in 10 patients with tuberculous meningitis in both acute phase and recovery phase,10 purulent meningitis,10 cryptococeal meningitis,10 viral encephalitis and 10 controls.The differences among the groups were compared by t test. The correlations between MMP-9 levels and the cell count,glucose, chloride, and protein in CSF were analyzed by Spearmanrank correlation.ResultsThe CSFMMP-9levelsingroupsoftuberculousmeningitis, purulentmeningitis,cryptoeoccal meningitis and viral encephalitis were (569.46±162.42),(182.79±99.06),(54.69±19.93) and (18.52±10.31) ng/mL,respectively,which were all significantly higher than that in controls (3.51± 1.53) ng/mL. There were significant differences between tuberculous meningitis group and other groups (t=2.925,3.041,3.237,3.454;P0.0340,0.0270,0.0080 and0.0001,respectively). Moreover,in tuberculous meningitis group,the MMP-9 level in acute phase was (569.46±162.42) ng/mL,which was significantly higher than that in recovry phase (294.30+89.06) ng/mL.The CSF level of MMP-9 in tuberculous meningitis group was positively correlated with CSF protein (r=0.509,P=0.044),negative correlated with CSF glucose (r=-0.451,P=0.008) and chloride (r=-0.637,P=0.007),but no correlation with CSF cell count (r=0.308,P=0.246). And there were no correlations in other groups. ConclusionsMMP-9 level in CSF increases significantly in patients with tuberculous meningitis and purulent meningitis,which can be used as a marker of CNS infection.Dynamic monitoring of thc CSF level of MMP-9 may be meaningful for the diagnosis and treatment.

Objective To investigate the changes of plasma levels of glucagon in cirrhotic patients and its effect in the pathogenesis of portal hemodynamics.Methods Plasma levels of glueagon were measured by radioimmunoassay(RIA)in 42 patients with liver cirrhosis and 20 healthy controls.The max diameters,mean flow velocity,flow rate of the portal vein trunks(PV)and splenic veins(SV)were detected by Color Doppler Ultrasound.Results Plasma levels of glucagon in cir- rhotic patients were significantly higher than those in controls and were increased with the severity of hepatic function im- pairment as assessed by Child-Pugh grade.Patients with ascites showed signifieanthigher plasma glucagon levels than those without ascites.Plasma levels of glueagon were positively correlated with portal vein diameters,splenic vein diame- ters and splenic vein flows.Conclusion The increase of plasma glucagon levels reflects,in part,the severity of impair- ment of hepaticfunction.In addition,it may contribute to the pathogenesis of portal hypertension.

OBJECTIVE To investigate the clinical characteristics,risk factors and preventive measures for nosocomial pneumonia in patients with post-hepatitis liver cirrhosis.METHODS A prospective and retrospective study was carried out to investigate the clinical data of 495 patients with post-hepatitis liver cirrhosis in Department of Infectious Diseases during Jan 1,2005 to Dec 31,2007.RESULTS The incidence rate of the nosocomial pneumonia in post-hepatitis liver cirrhosis patients was 13.50 %.The death rate was 25.40 %,which was obviously higher than 6.8% of patients without no nosocomial infection(?2=23.77,P

Objective To understand the epidemiological characteristics and clinical features of Zika virus disease,and to improve its prophylaxis and treatment.Methods The first case with imported Zika virus disease in China was retrospectively reported and analyzed.The literature of Zika virus infection in human was reviewed.Results This patient was the first case with imported Zika virus disease in China who presented with typical clinical characteristics and had clear epidemiological history.All the contacts were test negative for Zika virus nucleic acid.Literature retrieval showed evidence of Zika virus propagation in more than 40 countries in Africa,Asia,and Americas.The majority of patients presented with mild symptoms and the main prevention measures included mosquito control and improved awareness of personal protection.Conclusions Human infected with Zika virus often shows recessive infection.Only a small part develop disease and have generally good prognosis with supportive treatment.

Objective To investigate the incidence and risk factors of acute kidney injury (AKI)in hepatitis B virus (HBV)related acute-on-chronic liver failure (ACLF)patients,and to explore the impact of AKI on the prognosis of ACLF.Methods The medical records of 227 patients who were diagnosed with HBV-related ACLF at the Department of Infectious Diseases in the First Affiliated Hospital of Nanchang University from January 2015 to August 2016 were retrospectively reviewed.Patients were divided into AKI group and non-AKI group based on the AKI criteria published by International Club of Ascites in 2015 .Demographic and clinical data were compared between groups.The AKI incidence and its impact on patients’prognosis were analyzed.The comparison of continuous variables was done by t test or rank-sum test.The comparison of categorical variables was done byχ2 test or Fisher exact test.AKI risk factors were analyzed by using logistic regression.Results There were 66 (29.1 %)cases were diagnosed with AKI among 227 ACLF patients,among which,45 patients (68.2%)were stage Ⅰ,14 (21 .2%) were stage Ⅱ and 7 (10.6%)were stage Ⅲ.Age,cirrhosis,concentrations of total bilirubin and albumin,international normalized ratio (INR),percentage of neutrophils,MELD scores and spontaneous peritonitis rate (SBP)were all statistically different between AKI group and non-AKI group (all P <0.05).The binary logistic regression analysis revealed that only INR (OR=3.132,P =0.001 )and SBP (OR=4.204,P =0.001 )were the independent risk factors of AKI.The optimal cut-off value for INR was 2.025 with AUROC of 0.609 (P =0.01),sensitivity of 59.1 % and specificity of 62.1 %.The 30-day mortality of AKI group was significantly higher than non-AKI group (χ2= 18.324,P < 0.01). Conclusions AKI is relatively common in patients with ACLF.The risk factors of AKI are INR and SBP. AKI has significant impact on the short-term survival rate of ACLF.Therefore,physicians should pay attention to patients with INR of ACLF at admissions and SBP during the management so as to prevent the occurrence of AKI and to reduce the fatality of ACLF.

Objective To analyze the characteristics and significance of matrix metalloproteinase-9 (MMP-9) expression in the pathophysiological processes of tuberculous meningitis in mice.Methods Sixteen mice were intracerebroventricularly injected with H37RV suspension as the model group.Meanwhile,the other 16 mice were injected with 0.9％ sodium chloride solution as the control group.Thirty days later,all mice were decapitated and the brain tissue were respectively used to for Mycobacterium tuberculosis (M.tuberculosis) incubation,pathological changes observation,MMP-9 activity detection by zymography,blood-brain-barrier permeability and moisture content detection,and immunofluorescence stain of MMP-9,glial fibrillary acidic protein (GFAP) and integrin αM (OX-42).The t test was used to compare the differences between the two groups.Results Every experimental mouse was injected with (1.271±0.111) × 106 colony-forming units (cfu) M.tuberculosis.Thirty days later,the amount of M.tuberculosis in brain tissue homogenates was (4.900± 1.407) × 104 cfu/mL,and the hematoxylin and eosin staining showed dilatation of subarachnoid and ventricular and infiltration of a large number of inflammatory cells.The cumulative absorbance (A) of MMP-9 bands of brain tissue was 47 821 ± 19 932 in the model group and 10 082 ± 3 544 in the control group.The difference was statistically significant (t =3.728,P=0.010).The evans blue (EB) content of brain tissue was (11.8 ± 3.6) μg/g in model group and (4.7 ±3.4) μg/g in control group.The difference was statistically significant (t=2.887,P=0.028).The moisture of brain tissue was 0.849±0.035 in model group and 0.775±0.037 in control group.The difference was statistically significant (t=2.925,P=0.026).The immunofluorescence staining showed that the infected brain tissue expressed high degrees of MMP-9,GFAP and OX-42.And MMP-9 was overlapped with both GFAP and OX-42 obviously.Conclusions The activity of MMP-9 is significantly enhanced in brain tissue of mice suffering from tuberculous meningitis and participates in blood-brain barrier damage,tissue edema and inflammatory cells exudation.Microglia cells-astrocytes network is involved in the secretion of MMP-9.

Objective To study the impact of tumor necrosis factor-α (TNF-α) and its antagonist on the expressions of intestinal mucosa claudin-1,Zonula Occludens-1 (ZO-1) and myosin light chain kinase (MLCK) in rat models of acute liver failure.Methods Fifty four healthy male SpragueDawley (SD) rats were randomly divided into normal control group,model group and intervention group according to a random number table.Rats in normal control (n=6) group were intraperitoneally injected with 0.9％ saline (12 mL/kg).Rats in model group (n=24) and intervention group (n=24) were intraperitoneally injected with a full dose of D-galactosamine (D-GalN) at a dose of 1 200 mg/kg to establish model of acute liver failure,while rats in intervention group were intraperitoneally injected with TNF-α antagonists (rhTNFR∶Fc) at a dose of 12.5 mg/kg before 24 hours given D-GalN.At each time point of hour 8,24,48 and 72,six rats in both model group and the intervention group were sacrificed,respectively,while the normal control group were all anesthetized and sacrificed at 72 h.Models were repeated five times.Serum liver function was detected by biochemical method,and serum TNF-α level was detected by enzyme-linked immunosorbent assay (ELISA).Hematoxylin-eosin (HE) stained sections of liver and terminal ileum were examined under an optical microscope for pathological changes;and protein expression of the terminal ileum Claudin-1,ZO-1 protein and MLCK were determined by immunohistochemistry and Western blot.Means among groups were compared with t test.Results Acute liver failure was successfully induced in the D-GalN injected rats.In the model group,alanine aminotransferase (ALT) began to decline,total bilirubin continued to rise,and enzyme-jaundice separation developed at hour 72.But total bilirubin in intervention group at hour 72 was decreased.Light microscope showed that at hour 72,villus lodged at terminal ileum in the model group with part of villus tip failing off in the model group.Villus mucosa and submucosa interstitial were edema and infiltrated with numerous neutrophils.The terminal ileum kept integrate in the intervention group,and villus mucosa and submucosa were mild edema and only infiltrated with a small amount of neutrophil.Expressions of tumor necrosis necrosis factor (TNF)-α in rats of model group and intervention group were gradually increased and peaked at hour 24 ([239.83 ± 15.81] and [182.71± 17.08] ng/L,respectively),which were significantly higher than that of the control group ([24.19±3.57] ng/L,t=22.68and 15.73,respectively;both P＜0.01).Expression of serumTNF-α in the intervention group was significantly lower than that of model group (t=4.58,P＜0.01).Expressions of Claudin-1 and ZO-1 in model group decreased gradually at an early stage and reached the lowest level at hour 24 (0.355 ± 0.068 and 0.387 ± 0.091,respectively),which were both significantly lower than that of control group (1.640±0.188 and 1.015±0.150,respectively;t=12.87 and 7.14,respectively;both P＜0.01).In the intervention group,expressions of Claudin-1 and ZO-1 also decreased to the lowest level at hour 24 (1.051 ± 0.370 and 0.642 ± 0.082,respectivley),which were both significantly lower than that of control group (t =2.84 and 4.36,respectively;both P＜0.05),but significantly higher than model group with stastically difference (t =3.70 and 4.15,respectively;both P＜0.01).MLCK protein levels in the model and intervention group were gradually increased,which peaked at hour 24 (1.298±0.194 and 1.033 ± 0.073,respectively),significantly higher than the control group (0.460±0.069,t=8.16 and 11.44,both P＜0.01);and MLCK in the intervention group was lower than that in the model group with statistically difference (t=2.56,P＜0.05).Conclusions Expression of serum TNF-α in rat model of acute liver failure increases,which leads to decreased expression of Claudin-1 and ZO-1,and increased expression of MLCK,makes cell shrunk and cell gap increased.TNF-α antagonist could significantly reduce the inflammation and liver cell apoptosis,improve liver function by inhibiting MLCK expression and preventing decrease of Claudin-1 and ZO-1 proteins.

Objective:To investigate the predictive value of serum soluble urokinase-type plasminogen activator receptor (suPAR) combined with alpha-fetoprotein (AFP) and model for end-stage liver disease (MELD) score in short-term prognosis assessment of patients with chronic hepatitis B (CHB) related acute-on-chronic liver failure (ACLF).Methods:From January 2018 to May 2020, 66 patients with CHB related ACLF from Fuzhou First People′s Hospital were enrolled. After 90 days of follow-up, the patients with CHB related ACLF were divided into death group and survival group according to the outcome. Meanwhile, 30 patients with CHB were enrolled by simple random sampling method. The differences of serum suPAR in patients with CHB related ACLF and patients with CHB were analyzed. The values of suPAR, AFP and MELD score were compared between death group and survival group in patients with CHB related ACLF. The predictive value of suPAR, AFP, MELD score, Child-Turcotte Pugh score (CTP score) and suPAR combined with AFP and MELD score in the short-term prognosis of patients with CHB related ACLF were analyzed by area under the receiver operator characteristic curve (AUROC). Data were analyzed by two independent sample t test or non-parametric test. Results:The serum suPAR level of patients with CHB related ACLF was (9.6±0.8) ln ng/L, which was higher than that of patients with CHB ((8.0±0.3) ln ng/L). The difference was statistically significant ( t=14.533, P<0.01). The suPAR and MELD score of patients with CHB related ACLF in the death group were (9.9±0.7) ln ng/L and 29.6 (7.1) points, respectively, which were higher than those in the survival group ((9.4±0.7) ln ng/L and 21.0 (5.0) points, respectively). The AFP level in the death group was 45.9 (108.1) μg/L, which was lower than that in the survival group (209.3 (187.1) μg/L). There were significant differences in suPAR ( t=2.895, P=0.005), MELD score ( Z=4.708, P<0.01) and AFP ( Z=3.051, P<0.01) between the death group and the survival group. AUROC of suPAR (0.741, P=0.001), AFP (0.724, P=0.002) and MELD score (0.885, P<0.01) had predictive value for death in patients with CHB related ACLF. The sensitivities of suPAR, AFP, MELD score, CTP score and suPAR combined with AFP and MELD score were 84.6%, 73.1%, 88.5%, 96.2% and 84.6%, respectively, and the specificities were 75.0%, 72.5%, 70.0%, 52.5% and 92.5%, respectively. The AUROC of suPAR combined with AFP and MELD score was 0.871 ( P<0.01), which was higher than that of CTP score (0.793, P<0.01). Conclusions:Serum suPAR is increased in patients with CHB related ACLF. SuPAR combined with AFP and MELD score could apply in the prognostic value for patients with CHB related ACLF.

Objective: To explore the efficacy of glucocorticoid (GC) therapy on hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF), and the effectiveness and safety of voriconazole (VCZ) in preventing pulmonary Aspergillus infection in HBV-ACLF patients treated with GC. Methods: Two hundred and thirty-two patients with HBV-ACLF were enrolled from January 2016 to December 2018 in the First Affiliated Hospital of Nanchang University. They were divided into non-GC group (104 cases), GC group (74 cases), and GC+ VCZ group (54 cases). The observation period was four months. The baseline liver function, the incidence of pulmonary Aspergillus infection, the survival rate during observation period, and the incidence of complications were compared among the three groups. The adverse reactions of VCZ were observed to identify the best dose for prevention. Quantitative data were analyzed by analysis of variance or rank sum test. Count data were analyzed by chi-square test or Fisher exact test. Results: The baseline liver functions were not significantly different among the three groups (all P>0.05). The incidence of pulmonary Aspergillus infection in the GC group (22.97%(17/74)) was both higher than that in the non-GC group (5.77%(6/104)) and GC+ VCZ group (1.85%(1/54)), the differences were both statistically signifrcant (χ²=11.373 and 9.843, respectively, both P<0.01). The overall mortality rate of HBV-ACLF patients with pulmonary Aspergillus infection was 79.2%(19/24). The survival rate in non-GC group (37.5%(39/104)) showed no statistical difference with that in GC group (39.19%(29/74), χ²=0.052, P=0.819). The survival rate of GC+ VCZ group (66.67%(36/54)) was significantly higher than that in GC group and non-GC group (χ² =12.126 and 9.431, respectively, both P<0.01). The blood concentrations of VCZ were randomly measured in 16 patients from the GC+ VCZ group, and the range was 0.82-5.38 mg/L, with no evident adverse reactions. Conclusions The GC treatment is effective in HBV-ACLF patients in early stage. The VCZ treatment effectively reduces the incidence of pulmonary Aspergillus infection in HBV-ACLF patients receiving GC treatment and increases the survival rate. Oral VCZ (200 mg/d) treatment has a stable blood concentration in HBV-ACLF patients, with rare adverse reactions and good safety.