Objective To detect the serum viral load,lymphocyte subsets,serum enzymes and blood cell counts of severe fever with thrombocytopenia syndrome (SFTS) patients,and to use logistic regression analysis and receiver operating characteristic (ROC) curve to establish a model to analyze the severity of SFTS.Methods A case-control study of 24 SFTS cases admitted between May 2011 and July2012 was conducted at the First Affiliated Hospital of Nanjing Medical University.All SFTS cases were defined according to Fever with Thrombocytopenia Syndrome Prevention and Control Guidelines (2010 edition) issued by the Ministry of Health of the People's Republic of China.According to their disease severity,the patients were divided into two groups,the non-severe group (16 cases) and the severe group (8 cases).In addition,32 healthy volunteers were also enrolled in this study.Flow cytometry was adopted to detect the CD3 +,CD4 +,CD8 + lymphocytes and CD3-CD16 + CD56 + natural killer cells (NK cells) in the peripheral blood of SFTS patients,and cytometric beads array (CBA) was used to detect Th1/Th2/ Th17 cytokines.The serum viral load in patients with severe fever with thrombocytopenia syndrome virus (SFTSV) infection was detected by fluorescent quantitative PCR technology.Besides,white blood cells,platelets and serum enzymes were measured.The multi-index conjunctive model of logistic regression analysis and receiver operating characteristics (ROC) curve were used to analyze the predictive values of indexes on severity of SFTS.Results The ROC analysis of single index found that SFTSV RNA,CD3,CD4,CD8,CD56,AST,LDH,CK,IL-6 and IL-10 have good predictability on severity of SFTS in the early course of the disease ; the area under the curve (AUC) were 0.83,0.84,0.90,0.75,0.94,0.73,0.78,0.87,0.74 and 0.77 respectively,and the cut-off values of the SFTSV load,CD3 +,CD4 +,NK cells and CK were 6.19 log10 copies/ml,57.51％,19.47％,15.71％ and 696.45 U/L respectively.Using the step-by-step method logistic regression analysis to build a model and analyze the ROC curve of prediction probability,it was found that the predictability of joint indexes of SFTSV RNA/CD3/CD4/CD8/CD56,SFTSV RNA/ AST/lactate dehydrogenase (LDH)/CK,or SFTSV RNA/IL-6/IL-10 increases in a certain degree.The areas under the ROC curve were 0.95 (95 ％ CI:0.00-1.00),0.87 (95 ％ CI:0.75-0.99),and 0.83 (95 ％ CI:0.70-0.97),the sensitivities were 93.3％,86.70％ and 77.30％,and the specificities were 94.4％,83.30％ and 83.30％ respectively.Conclusions The level of serum SFTSV is highly related to the severity of the disease.The high levels of serum SFTSV load,LDH,CK,IL-6,and IL-10,together with significant reduction of CD3 + cells and CD4 + cells,may indicate poor prognosis of the SFTS patients.Based on the logistic regression model by multiple indexes,the severity of SFTS can be better predicted.

OBJECTIVE:To study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).METHODS:PBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.RESULTS:Exposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).CONCLUSION:HBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.

This report aims to investigate the dynamical changes of HBcAg18-27 epitope specific cytotoxic T lymphocytes(CTL), alanine aminotransferase (ALT), HBV DNA and HBsAg in peripheral blood of acute hepatitis B patients, and to explore the roles of HBcAg18-27-specific CTLs in virus clearance and liver injury. Acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients were divided into two groups according to results of HLA-A0201. Patients with positive HLA-A0201 were classified into HBcAg-specific CTL group and those with negative HLA-A0201 were referred as control group. The specific CTLs were stained with HLA-A0201 limited HBcAg18-27 epitope MHC-Pentamer and the frequencies of CTLs, T, B, NK and NKT cells were detected by flow cytometry (FCM). The serum ALT, HBV DNA and HBsAg were examined using speed analysis, quantitative PCR and abbott chemiluminescent technology. The frequencies of HBcAg18-27-specific CTLs in AHB patients were higher in the early three weeks as compared to the late three weeks. The apex time of HBV-specific CTL frequencies lagged behind those of HBV DNA, HBsAg and ALT. The loss of HBsAg in patients with high frequencies of HBV-specific CTL was earlier than that in patients with low frequencies (t = 2.018, P value is less than 0.05). In the second week the peak frequencies of CD3+CD8+ cells overlapped with that of HBcAg18-27-specific CTLs and with a positive correlation between (r = 0.420, P value is less than 0.05). During the early stages of AHB, the frequencies of NK and NKT cells were found significantly lower than that of control group and CHB group and the levels were back to normal after recovery. Moreover, a negative correlation existed between the frequencies of NK cells and the dynamic changes of HBcAg18-27-specific CTLs (r = -0.435, P value is less than 0.01) in AHB group. The frequencies of HBcAg18-27-specific CTLs were significantly higher as compared to CHB group in the first three weeks (z = -3.258, -4.04, and -3.259, P value is less than 0.01). The early loss of HBsAg was closely related to the high frequencies of HBcAg18-27 specific CTLs in AHB patients. HBcAg-specific CTL frequencies in peripheral blood could be used to predict clinical outcome after HBV infection. The frequencies of CD8+ T cells can reflect the changes of frequencies of HBcAg-specific CTL during acute HBV infection.

To investigate the clinical feature, risk factors and outcome of treatment in patients with liver failure complicated by invasive fungal infections.Retrospective analysis of the clinical data and related factors of 27 patients with liver failure complicated by invasive fungal infections was performed.These patients were admitted from January 2007 to August 2009 in our department.Among them, Candida albicans accounted for 17cases (54.84%), albicans tropicals for 4 cases (12.90%).Fungal infection in respiratory tract and alimentary tract accounted for 58.06% and 11% respectively.81.25% of them had fever fluctuating from 37.4 o C to 40 o C. 81.25% had elevated white blood cell counts .All had the usage of broad-spectrum of antibiotics, whereas some of them used corticosteroids and had invasive medical manipulation for the treatment.Most patients deteriorated after invasive fungal infections.21 cases accepted with the treatment of antifungal drugs and mortality rate was 63.00%.It was found that the invasive fungal infection possibility of patients with liver failure significantly increased.To prevent the occurrence of invasive fungal infection, promptly early treatment of liver failure, proper use of antibiotics, cautious use or disuse of corticosteroids, reduction of invasive medical manipulation should be well done.Early detection and treatment of fungal infection are vital to decrease in mortality rate.

OBJECTIVES In order to investigate the relationship among the toll-like receptor 2 (TLR2), hepatitis B e antigen and HBV DNA, the expression levels of TLR2 on peripheral blood monocytes of chronic hepatitis B (CHB) patients as well as on their monocytes stimulated by ligand of TLR2 (Pam3CSK4) and HBeAg were analyzed. METHODS Sixty-eight adults with CHB were enrolled, including 37 HBeAg-positive patients, 17 HBeAg-negative and HBV DNA negative patients, and 14 HBeAg-negative and HBV DNA positive patients. Sixteen healthy volunteers were also studied as controls. TLR2 expression levels on their peripheral blood monocytes stimulated with Pam3CSK4 or not stimulated were analyzed by FACS Caliber. The relationship of the expression levels of TLR2, HBeAg and HBV DNA were also analyzed. The level of TLR2 on peripheral blood monocytes of healthy volunteers and HBeAg-negative CHB patients stimulated by HBeAg was examined for six hours. RESULTS The TLR2 expression levels on CD14+ cells were significantly reduced in HBeAg-positive patients (47.57%+/-21.40 %) compared to both healthy volunteers (76.51%+/-7.46%) and HBeAg-negative patients (HBV DNA positive group 73.2%+/-14.2%, HBV DNA negative group 75.2%+/-11.3%); but there was no difference between those of the HBeAg-negative patients and the healthy volunteers. Expression levels of TLR2 on monocytes stimulated by TLR2 ligand in HBeAg-positive patients were obviously increased, and reached the basic levels of the healthy volunteers and the HBeAg-negative patients. The expression levels of TLR2 on monocytes stimulated by HBeAg of the healthy volunteers and the HBeAg-negative patients were markedly reduced. CONCLUSIONS In the presence of HBeAg, HBV down-regulates the expressions of TLR2 on CD14+ cells from peripheral blood, and there is no correlation between HBV-DNA and TLR2. Pam3CSK4 can boost the TLR2 expression in HBeAg-positive patients. The proposed interaction between HBV and TLR2 may provide an important clue to explain the reasons of the establishment of persistent HBV infection.