Objective To summarize the causes of patients with fever of unknown origin (FUO) and to analyze the relationship between infectious diseases and FUO,in order to provide envidence for experiential therapy.Methods Clinical data of 374 FUO inpatients at He'nan Provincial People's Hospital from June 1,2009 to May 31,2013,including gender,age,diagnosis and department were analyzed retrospectively.Results Three hundred and twenty-seven cases among the overall 374 FUO patients (87.4％) were eventually etiological diagnosed based on supplementary examinations or diagnostic treatment.As for the causes of fever,209 were infection,accounting for 55.9％,among which 78 cases (20.9％) were diagnosed with tuberculosis,23 cases (6.1％) brucellic diseases,19 cases (5.1％) rickettsia infection.Meanwhile,the noninfectious diseases,such as connective tissue diseases (47,12.6％),hematonosis (37,9.9％) as well as the solid tumors (13,3.5％) also constituted considerable shares of the causes for FUO.However,the causes of 47 cases (12.6％) were not identified before discharge.Conclusions Infectious diseases are the main cause of FUO,in which tuberculosis accounts for the majority.Brucellosis and rickettsia infection also account for a considerable proportion.The causes of most FUO cases could be identified through detailed analysis of clinical data and supplemental examinations.

Objective:To observe the level and detection of ascites CD100 on the activity of CD4 + and CD8 + T lymphocytes in vitro in the peripheral blood of patients with liver cirrhosis combined with spontaneous bacterial peritonitis. Methods:Peripheral blood and ascites were collected from 77 cases of liver cirrhosis (49 patients with liver cirrhosis combined with simple ascites and 28 patients with liver cirrhosis combined with SBP), and peripheral blood was collected from 22 controls. Soluble CD100 (sCD100) in peripheral blood and ascites was detected by an enzyme-linked immunosorbent assay. Flow cytometry was used to detect membrane-bound CD100 (mCD100) on the surface of CD4 + and CD8 +T lymphocytes. CD4 + and CD8 +T lymphocytes in ascites were sorted. CD4 +T lymphocyte proliferation, key transcription factor mRNA, and secreted cytokine changes, as well as CD8 +T lymphocyte proliferation, important toxic molecule mRNA, and secreted cytokine changes, were detected after CD100 stimulation. The killing activity of CD8 +T cells was detected by direct contact and indirect contact culture systems. Data conforming to normality were compared using one-way ANOVA, a student's t-test, or a paired t-test. Data that did not conform to a normal distribution were compared using either the Krusal-Willis test or the Mann-Whitney test. Results:There was no statistically significant difference in plasma sCD100 level between patients with liver cirrhosis combined simple ascites (1 415 ± 434.1) pg/ml, patients with liver cirrhosis combined with SBP (1 465 ± 386.8) pg/ml, and controls (1 355 ± 428.0) pg/ml ( P = 0.655). The ascites sCD100 level was lower in patients with liver cirrhosis combined with SBP than that of patients with simple ascites [(2 409 ± 743.0) pg/ml vs. (2825±664.2) pg/ml, P=0.014]. There was no statistically significant difference in the level of mCD100 in peripheral blood CD4 + and CD8 + T lymphocytes among the three groups ( P > 0.05). The levels of mCD100 in ascites CD4 + and CD8 + T lymphocytes were higher in patients with liver cirrhosis combined with SBP than those in patients with simple ascites ( P < 0.05). CD100 stimulation had no significant effect on the proliferation of CD4 + and CD8 +T lymphocytes in the ascites of patients with liver cirrhosis combined with SBP ( P > 0.05). There were no significant effects on the expression of transcription factors in effector CD4 +T lymphocytes (T-bet, retinoic acid associated solitary nuclear receptor γt, aromatic hydrocarbon receptor) or secretion of cytokines (interferon-γ, 17, and 22) ( P > 0.05). CD100 stimulation had increased the relative expression of perforin, granzyme B, and granlysin mRNA and the levels of secreted interferon-γ and tumor necrosis factor-α, killing activity in ascites CD8+ T lymphocytes of patients with liver cirrhosis combined with SBP ( P < 0.05). Conclusion:The active form of CD100 is sCD100 instead of mCD100. There is an imbalance between the expression of sCD100 and mCD100 in the ascites of patients with cirrhosis combined with SBP. sCD100 can enhance the function of CD8 +T lymphocytes in the ascites of patients with cirrhosis combined with SBP and thus is one of the potential therapeutic targets.

Objective To explore the molecular mechanisms of hepatic stimulator substance (HSS) gene knockout in promoting the development and progression of nonalcoholic steatohepatitis (NASH).Methods NASH model mice (n=20) with HSS wild-type (HSS+/+) or HSS gene knockout (HSS-/-) were constructed using modified choline-deficient diet (CD-diet),untreated C57BL6-HSS-/-and C57BL6-HSS+/+ mice (n=20) were considered as control.Ten mice of each group were killed at month 1 and 2,respectively.The levels of triglyceride (TG) and total cholesterol (TC) in liver were measured using ELISA method.Histopathology and collagen deposition in liver tissue were observed using HE staining and Masson staining,respectively.Lipid content in liver tissue was observed and calculated using oil red O staining.The levels of mRNA and proteins of peroxisome proliferators activated receptor gama coactivator 1 alpha (PGC-1α),mitochondrial transcription factor A (TFAM),transcription factor-E2 related factor α (Nrf2),[-loop,dynamin-related protein 1 (Drp1),mitochondrial fission 1 protein (Fis1),mitofusins 1 (Mfn1),autophagy related gene 3 (Atg3) in liver tissue were detected using Real-time PCR and Western blot,respectively.Content of malonaldehyde (MDA),cyclooxygenase Ⅳ (COX Ⅳ) and adenosine tirphosphate (ATP) were measured using kits,and the activity of respiratory chain complex Ⅴ and cytochrome C oxidase in liver tissue were measured using spectrophotometry.the comparison between groups was done by t test.Results The levels of HSS mRNA and protein in mice-HSS-/-were 0.154± 0.04 and 0.08± 0.01,respectively,which were both significantly lower than those in mice-HSS+/+ (0.952 ± 0.08 and 1.362±-0.130,respectively),and t he differences had statistical significance (t =10.244 and 10.375,respectively,both P＜0.05).One month and 2 months after NASH modeled,TC contents in mice-HSS-/ were (248.6±21.5) μmol/g and (217.4±18.0) μmol/g,respectively,which were both remarkably higher than those in mice-HSS+/+ [(153.5 ± 11.2) μmol/g and (140.8 ±7.5) μmol/g,respectively],and the differences had statistical significance (t=15.270 and 10.524,respectively,both P＜0.05).The results form HE staining,oil red O staining and Masson staining indicated that fat deposition,collage deposition and inflammation in liver tissues of mice-HSS-/-were severer than those in mice-HSS+/+.One month after NASH modeled,protein levels of Drp1,Fis1,Mfn1 and Atg3 in liver tissues of mice-HSS-/ were all significantly decreased compared with those in mice-HSS+/+,and the differences had statistical significance (t=10.705,24.072,9.892 and 17.540,respectively,all P＜ 0.05).Two months after NASH modeled,protein levels of Drp1,Fis1,Mfn1and Atg3 in liver tissues of mice-HSS-/ were all significantly decreased compared with those in mice-HSS+/+,and the differences had statistical significance (t=125.378,15.926,34.330 and 13.437,respectively,all P＜0.05).One month after NASH modeled,mRNA levels of Drp1,Fis1,Mfn1 and Atg3 in liver tissues of mice-HSS-/-were all significantly decreased compared with those in mice-HSS+/+,the differences had statistical significance (t=36.337,40.825,33.508 and 28.104,respectively,all P＜0.05).Two months after NASH modeled,mRNA levels of Drp1,Fis1,Mfn1 and Atg3 in liver tissues of mice-HSS-/-were all significantly decreased compared with those in mice-HSS+/+,and the differences had statistical significance (t=35.210,42.375,27.753 and 20.560,respectively,all P＜0.05).The protein levels of PGC-1α,TFAM,Nrf2 and D-loop in liver of C57BL6-HSS-/-group were lower than those in liver of C57BL6-HSS+/+ group,and the differences had statistical significance (one month:t=20.548,31.036,19.445 and 10.974,respectively;two months:t=9.887,13.330,22.375 and 18.903,respectively,all P＜0.05).The mRNA levels of PGC-1α,TFAM,Nrf2 and D-loop in liver of C57BL6-HSS-/-group were all lower than those in C57BL6-HSS+/+ group,and the differences had statistical significance (one month:t=9.087,12.582,21.451 and 7.774,respectively;two months:t=23.758,17.924,9.924 and 15.209,respectively,all P＜0.05).One month and 2 months after NASH modeled,the levels of ATP mRNA in liver of C57BL6-HSS / group were both significantly lower than those in C57BL6-HSS+/+,and the differences had statistical significance 0=43.775 and 28.375,respectively,both P＜0.05);the levels of COXⅣ mRNA in liver of C57BL6-HSS / group were 0.142 ± 0.06 and 0.068± 0.001,respectively,which were both significantly lower than those in C57BL6-HSS+/+ group (0.255± 0.08 and 0.172 ±0.06,respectively),and the differences had statistical significance (t=28.337 and 19.782,respectively,both P＜0.05);the levels of MDA mRNA in liver of C57BL6-HSS-/-group were 0.973 ±0.112 and 1.253±0.054,respectively,which were both significantly lower than those in C57BL6-HSS+/+ group (0.366±0.02 and 0.872±0.05,respectively),and the differences had statistical significance (t=8.357 and 6.582,respectively,both P＜0.05).Conclusion Deletion of HSS accelerates NASH progression via inhibiting mitochondrial fusion,which leads to dysfunction of mitochondrial respiratory chain and inhibition of fatty acid oxidation.

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.

Background: Enteritis is one of the most frequently reported symptoms in piglets infected with porcine circovirus type 2 (PCV2), but the immunopathogenesis has not been reported. Objectives: This study examined the effect of a PCV2 infection on the intestinal mucosal immune function through morphological observations and immune-related molecular detection. Methods: Morphological changes within the ileum of piglets during a PCV2 infection were observed. The expression of the related-molecules was analyzed using a gene chip. The immunocyte subsets were analyzed by flow cytometry. The secretory immunoglobulin A (SIgA) content was analyzed by enzyme-linked immunosorbent assay. Results: The PCV2 infection caused ileal villus damage, intestinal epithelial cells exfoliation, and an increase in lymphocytes in the lamina propria at 21 days post-infection.Differentially expressed genes occurred in the defense response, inflammatory response, and the complement and coagulation cascade reactions. Most of them were downregulated significantly at the induction site and upregulated at the effector site. The genes associated with SIgA production were downregulated significantly at the induction site. In contrast, the expression of the Toll-like receptor-related genes was upregulated significantly at the effector site. The frequencies of dendritic cells, B cells, and CD8 + T cells were upregulated at the 2 sites. The SIgA content decreased significantly in the ileal mucosa. Conclusions PCV2 infections can cause damage to the ileum that is associated with changes in immune-related gene expression, immune-related cell subsets, and SIgA production.These findings elucidated the molecular changes in the ileum after a PCV2 infection from the perspective of intestinal mucosal immunity, which provides insights into a further study for PCV2-induced enteritis.

Objective: To observe ascitic interleukin-7 expression level in cirrhotic patients complicated with spontaneous bacterial peritonitis, and to detect the effect of recombinant human IL-7 on CD4⁺ and CD8⁺T lymphocyte function. Methods: A total of 84 patients with liver cirrhosis who were hospitalized from August 2017 to April 2018 were selected. Among them, 51 cases were complicated with cirrhosis and untainted ascites, and 33 cases were cirrhosis complicated with spontaneous bacterial peritonitis. Peripheral blood and ascites were collected routinely. The levels of IL-7 in peripheral blood and ascites were measured by enzyme-linked immunosorbent assay. CD4⁺T cells and CD8⁺T cells were purified from ascites, and were stimulated with recombinant IL-7. Cellular proliferation, key transcription factors for mRNA, and cytokines production by CD4⁺T cells in response to IL-7 stimulation was measured. mRNA expression corresponding to perforin, granzyme B, and granulysin as well as cytokines production by CD8⁺T cells was also measured in response to IL-7 stimulation. Cytolytic and non-cytolytic activity of CD8⁺T cells in response to IL-7 stimulation was also investigated in both direct and indirect contact co-culture system. Measurement data of the normal distribution were compared between the two groups by Student’s t-test and the data before and after stimulation were compared by paired t-test. Measurements that did not conform to normal distribution were compared between the two groups using Mann-Whitney U test, and data before and after stimulation were compared using Wilcoxon paired test. Results: There was no significant statistical difference in serum IL-7 levels between the two groups [(5 001 ± 1 458) pg/ml vs. (4 768 ± 1 128) pg/ml, P = 0.41]. The level of ascitic IL-7 in cirrhotic patients complicated with SBP was significantly lower than cirrhosis patients with untainted ascites [(966.4 + 155.8) pg/ml vs. (792.1 + 126.4) pg/ml, P < 0.01]. Recombinant IL-7 stimulation promoted the proliferation of CD4⁺ and CD8⁺T cells from ascites in patients with liver cirrhosis complicated by SBP. T-bet mRNA relative expression and IFN-γ secretion in CD4⁺T cells was also elevated in response to IL-7 stimulation in vitro. Moreover, IL-7 stimulation also increased the mRNA expressions of perforin, granzyme B, and granulysin as well as productions of IFN-γ and TNF-α by CD8⁺T cells. Recombinant IL-7 stimulation elevated cytolytic and non-cytolytic activity of CD8⁺T cells from ascites in patients with liver cirrohosis complicated by SBP, which manifested as increased target cell death and IFN-γ production in both direct and indirect contact co-culture system. Conclusion Ascitic IL-7 promotes T lymphocyte function in patients with liver cirrhosis complicated with SBP.

Objective To explore the correlation between adiponectin and liver fibrosis in patients with chronic hepatitis B virus infection.Methods 60 patients with chronic hepatitis B were divided into mild liver fibrosis group (n =28),severe liver fibrosis group (n =17)and cirrho-sis group (n =15),and 20 healthy volunteers were designed as the control group.The serum adiponectin level was detected in all the groups.Results There were significant differences of serum adiponectin levels among the four groups (P <0.05),and there was a positive correlation be-tween serum adiponectin level and liver fibrosis grade (r =0.967,P <0.05).The expression rate of liver tissue adiponectin in cirrhosis group was 93.33%,which was significantly higher than the mild group (14.29%)and severe fibrosis group (41.18%)(P <0.05).Conclusion Adiponectin is closely related to the liver fibrosis in patients with chronic hepatitis B virus infection.

Objective To explore the correlation between adiponectin and liver fibrosis in patients with chronic hepatitis B virus infection.Methods 60 patients with chronic hepatitis B were divided into mild liver fibrosis group (n =28),severe liver fibrosis group (n =17)and cirrho-sis group (n =15),and 20 healthy volunteers were designed as the control group.The serum adiponectin level was detected in all the groups.Results There were significant differences of serum adiponectin levels among the four groups (P <0.05),and there was a positive correlation be-tween serum adiponectin level and liver fibrosis grade (r =0.967,P <0.05).The expression rate of liver tissue adiponectin in cirrhosis group was 93.33%,which was significantly higher than the mild group (14.29%)and severe fibrosis group (41.18%)(P <0.05).Conclusion Adiponectin is closely related to the liver fibrosis in patients with chronic hepatitis B virus infection.

Objective: To investigate the coagulation function in patients with acute-on-chronic liver failure (ACLF) patients using thromboelastography (TEG), and to comprehensively and dynamically evaluate patients’ bleeding and coagulation status. Methods: The clinical data of ACLF patients were collected, and TEG was used to evaluate whole blood clotting kinetics in these patients. Routine biochemical parameters were measured, and complications were evaluated. The t-test was used for comparison of continuous data, the chi-square test was used for comparison of categorical data, and the Pearson correlation coefficient was used for correlation analysis. P < 0.05 was considered statistically significant. Results: A total of 60 patients (39 male and 21 female patients) were enrolled, with a mean age of 47.20±16.20 years. The TEG results showed that all patients had normal thrombokinetics, but TEG parameters were correlated with coagulation function, markers of systemic inflammatory response syndrome, laboratory markers, and prognosis. The patients with a higher R value had higher risks of infection (6.23±2.91 vs 4.74±1.12, P = 0.009), hepatorenal syndrome (5.64±2.54 vs 3.21±1.43 P < 0.01), and bleeding (6.71±3.51 vs 4.80±1.63, P = 0.01), and the patients with a lower K value (0.72±1.36 vs 1.64±1.43, P = 0.02), an increased α-angle (63.33°±10.02° vs 56.62°±12.13°, P = 0.03), and an increased MA (56.83±11.07 vs 50.40±10.81, P = 0.03) had increased risks of hepatic encephalopathy. Conclusion ACLF patients have low coagulation function, and TEG truly reflects the "rebalance" status of this low level. Abnormal TEG parameters suggest increased risk of complications in these patients, indicating a poor prognosis.