Objective:To investigate the related factors and prognosis of invasive Klebsiella pneumoniae liver abscess syndrome (IKLAS). Methods:The in-patients diagnosed with Klebsiella pneumoniae liver abscess in the Department of Infectious Diseases, Huashan Hospital, Fudan University from January 2015 to February 2021 were retrospectively enrolled. The patients were divided into IKLAS group and non-IKLAS group according to whether they had IKLAS or not. The clinical data between the two groups were compared, including the prevalence of diabetes mellitus, the details of liver abscess, clinical symptoms such as fever and abdominal pain, as well as laboratory tests such as glycosylated hemoglobin and hemoglobin. Statistical analysis was performed using chi-square test or independent sample t test. Multivariate logistic regression analysis was used to analyze the factors influencing the occurrence of IKLAS. Results:A total of 75 patients with Klebsiella pneumoniae liver abscess were enrolled, including 55 patients (73.33%) in the IKLAS group and 20 patients (26.67%) in the non-IKLAS group. Fifty-two point seven three percent (29/55) of the patients had diabetes mellitus and 12.73%(7/55) of the patients had abdominal pain in the IKLAS group, which were 20.00%(4/20) and 45.00%(9/20) in the non-IKLAS group, respectively, and the differences were both statistically significant ( χ2=6.38 and 7.28, respectively, both P<0.05). Most of liver abscesses were single (50/75, 66.67%), and more likely to occur in the right liver (50/75, 66.67%). The maximum diameter of liver abscess in the IKLAS group was (4.58±2.04) cm, which was smaller than that in the non-IKLAS group ((6.49±3.11) cm), and the difference was statistically significant ( t=2.82, P=0.011). Compared with those in the non-IKLAS group, patients in the IKLAS group had higher glycosylated hemoglobin (8.69%±2.64% vs 6.18%±1.31%) and hemoglobin ((112.25±22.04) g/L vs (100.05±18.59) g/L), and the differences were both statistically significant ( t=-4.25 and -2.21, respectively, both P<0.05). The proportion of patients using antibiotics combined with abscess drainage in the IKLAS group was 38.18%(21/55), and that in the non-IKLAS group was 85.00%(17/20). The difference between the two groups was statistically significant ( χ2=12.86, P<0.001). A total of 16 patients (21 eyes) were diagnosed as endogenous Klebsiella pneumoniae endophthalmitis (EKPE), and all of them were IKLAS patients, and 14 patients underwent monocular/binocular eyeball injection and/or vitrectomy and silicone oil filling. The visual acuity of 13 patients decreased significantly. Multivariate logistic regression analysis showed that complicated with diabetes mellitus was an independent risk factor for IKLAS (odds ratio ( OR)=5.02, 95% confidence interval (95% CI) 1.01 to 25.03, P=0.049). The large diameter of liver abscess was a protective factor for IKLAS ( OR=0.64, 95% CI 0.47 to 0.86, P=0.003). Conclusions:The patients with IKLAS have less abdominal pain, and most of them complicate with diabetes mellitus. Diabetes mellitus is an independent risk factor for the occurrence of IKLAS, while the large diameter of liver abscess is a protective factor. EKPE is associated with poor visual prognosis.

Since the discovery of circulating hepatitis B virus (HBV) RNA in the peripheral blood of patients with chronic hepatitis B in 1996, a growing number of studies have focused on clarifying the biological characteristics and clinical application value of serum HBV RNA. This consensus mainly summarizes the research progress of serum HBV RNA existing profiles, quantitative detection methods, and current clinical applications. In order to better apply this indicator for the clinical management of patients with chronic HBV infection, recommendations on quantitative detection target regions, detection results, and clinical applications are put forward.

Objective:To analyze the changes and efficacy of antiviral treatment regimens in patients with chronic hepatitis C.Methods:This was a single center retrospective study. A total of 157 patients with chronic hepatitis C in Huashan Hospital, Fudan University from January 2014 to February 2019 were included. Clinical informations of antiviral treatment and follow-up were collected. The sustained virologic response (SVR) rate and adverse events in patients receiving different antiviral regimens were compared. Chi-square test was used for statistical analysis.Results:Among the 157 patients, 133 patients had sufficient follow-up data. Seventy-one patients received treatment before 2017, among which 63 patients received interferon regimens and the SVR rate was 74.65%(53/71). Sixty-two patients received treatment after 2017, among which 61 patients received direct-acting antiviral agents (DAA) regimens and the SVR rate was 98.39%(61/62). The difference in SVR rate between the two groups was statistically significant ( χ2=15.230, P<0.01). In 69 patients who received DAA regimens from 2014 to 2019, the SVR at post-treatment week 12 (SVR12) was 95.65%(66/69). Among 43 patients who received DAA regimens containing sofosbuvir, the SVR12 rates of patients with hepatitis C virus genotype 1, 3 and other genotypes were 15/15, 5/6 and 90.91%(20/22), respectively. All the 26 patients who received DAA regimens non-containing sofosbuvir achieved SVR12. The SVR12 rates of patients with different hepatitis C virus genotypes and DAA regimens were not significantly different ( χ2=5.243, P=0.263). The incidences of adverse events in pre-2017 group and post-2017 group were 84.62%(77/91) and 6.06% (4/66), respectively. The difference was statistically significant ( χ2=94.520, P<0.01). The most common adverse events were decreases in neutrophil cell count, decreases in hemoglobin level and decreases in platelet count. Treatment was ceased in six patients due to adverse events. Conclusions:After 2017, the majority of patients with chronic hepatitis C received DAA regimens instead of interferon regimens. The SVR rate increases and the incidence of adverse events decreases along with the changes of leading treatment regimens.The SVR12 rate is higher in patients receiving DAA regimens, regardless of hepatitis C virus genotypes.

Objective:To evaluate the clinical utility of a novel quantitative assay named hepatitis B virus (HBV) simultaneous amplification and testing (SAT) using a kit for HBV RNA detection (RNA probes).Methods:HBV RNA was detected in 170 serum samples of chronic hepatitis B patients and 30 serum samples of patients without HBV infection collected by simple random sampling method from June 2017 to June 2018 in Huashan Hospital, Fudan University, Shanghai using HBV SAT and reverse transcription polymerase chain reaction (PCR) method. The sensitivity, specificity, kappa value and quantitative correlation of the two methods were analyzed and compared.The detection rates of HBV RNA from samples with different HBV DNA concentrations of the two methods were analyzed and compared. Statistical analysis was performed by chi-square test.Results:Based on the clinical diagnosis, the detection sensitivity, specificity, kappa value of HBV SAT were 97.06%(165/170), 100.00%(30/30) and 0.908, respectively, while the reverse transcription PCR were 92.94%(158/170), 100.00%(30/30) and 0.798, respectively. Among samples with lower concentration of HBV DNA (HBV DNA<100 IU/mL), the detection rates of HBV SAT and reverse transcription PCR were 77.27%(17/22) and 59.09%(13/22), respectively. The linear correlation coefficient of the two methods was r=0.987 8. Conclusions:Quantitation results of HBV RNA by HBV SAT and reverse transcription PCR method are consistent. HBV SAT is a rapid and accurate method for HBV RNA quantitative detection, which has a slightly higher detection rate than reverse transcription PCR in samples with low concentration of HBV DNA.

Objective:To study the changes of cardiopulmonary function in patients with rheumatoid arthritis,and to analyze the correlation between the changes ofcardiopulmonary function with oxidative stress and the peripheral blood lymphocyte attenuation factor.Methods:130 cases of patients with rheumatoid arthritis were studied as case group, and 50 cases of healthy persons were studied as normal control group.Detected the heart function parameters of two groups,which contained EF%,SV%,FS%,E A,E/A;lung function parameters of FVC,FEV1,MVV,PEF;B and T lymphocyte attenuation factor expression and activation level.Peripheral Cytokine(IL-17 and TNF-α,IL-4,IL-35) and oxidative stress index (ROS,MDA,SOD,TAOC) were detected by enzyme linked immu-nosorbent assay.Results:The indexes of cardiac function in the case group were significantly lower than that in the control group.103 cases had abnormal cardiac function index in the case group,which accounted for 79.23% of the case group,while the E/A had the highest abnormal rate.The case group had thickening of LADd, increasing of peak A, decreasing of EF, E peak and E /A, than the normal control group,the differences were statistically significant (P<0.05).Compared with normal control group,pulmonary function parameters were significantly lower in case group.88 cases of case group had abnormal pulmonary function,accounting for 67.69% of the case group.Among them,the abnormal rate of PEF was the highest.Pulmonary function indexes of case group was significantly lower than that of the control group, the difference was statistically significant ( P<0.05 ).Correlation analysis showed that the correlation coefficient of cardiac function indexes EF with CD24+cells and CD19+CD24+cells were respectively -0.353 and -0.457,which had negative correlation,with ROS the correlation coefficient was 0.459,which had positive correlation.The correlation coefficient of the cardiac function indexes in FS with CD24+cells,and CD19+CD24+cells was -0.395 and -0.421,which had negative correlation; the correlation coefficient of peak A and CD19+cell was 0.423,which had positive correlation;the correlation coefficient of E/A and BTLA was 0.393,which had obvious positive correlation.SV and MDA,SOD were positively correlated.The parameters of lung function with hs-CRP and ESR had significantly negative correlation.The correlation coefficient of lung function parameters FVC with BTLA and CD19+CD24+were 0.513 and 0.596,which had a significant positive correlation,with the correlation coefficient and CD24+BTLA+, TNF-αwere -0.451 and-0.351,which had significantly negative correlation.The correlation coefficients of FEV1 with CD24+CD19+, TAOC and IL-4 were 0.535,0.466 and 0.519,which showed a positive correlation,with CD24+BTLA+,MDA were -0.461 and -0.358,which had significantly negative correlation.The correlation coefficient of PEF with SOD,TAOC,IL-4,IL-35 were 0.547,0.482, 0.643 and 0.452,which had significantly positive correlation,with MDA,ROS,IL-17 were -0.451,-0.423 and -0.417,which had a significant negative correlation ( P<0.05 ).Conclusion: RA imbalance of oxidative stress and cell immune disorders which runs through the whole process in the heart and lung injury.Therefore,in clinical treatment,treatment of joint symptoms in RA patients needs restore the body′s redox homeostasis,in order to increase the level of BTLA,activate B cell and,T cell,thereby inhibiting immune and inflammatory response,reducing the heart and lung function impairment.

Background and purpose:Overexpression of inhibitor of protein phosphatase 2 A-2 (I2PP2A) in many tumors including gastric cancer suggests that I2PP2A may contribute to the development of gastric cancer. To further study the biological function of I2PP2A and its role in gastric cancer, we established a BGC823 cell line for stable expression of shRNA targeting human I2PP2A gene. Methods: A double-stranded shRNA targeting the I2PP2A was designed, synthesized and was inserted into a lentivirus vector (pGLV2), and the insertion was identiifed by restriction endonuclease analysis and DNA sequencing. BGC823 cells were then transfected with the packaged recombinant lentivirus, and resistant cell clones were selected with puromycin. The expression of I2PP2A was examined using real-time PCR (RT-PCR) and Western blot. Results:Sequencing result proved that recombinant lentivirus vector pGLV2-shRNA-I2PP2A was constructed correctly. RT-PCR and Western blot results conifrmed that the expression of I2PP2A was signiifcantly down-regulated in this infected BGC823 cell line. The efifciency of siRNA interference of I2PP2A could be up to about 90%. Conclusion:A lentiviral vector carrying a shRNA targeting the I2PP2A gene is successfully constructed, and a BGC823 cell line stably expressing I2PP2A shRNA is established with this lentiviral system.

T cell immunoglobulin and mucin domain 3 (Tim-3) is well known to negatively regulate T cells responses, but its role in burn-induced T cells immune suppression remains unclear. In the present study, in order to identify the relationship between Tim-3 expression and post-burn T cells immune suppression, C57BL/6 mice were subjected to burn injury or sham injury, and the liver and spleen were harvested at the day 1 after operation. The expression level of Tim-3 on hepatic or splenic T cells and the functional properties of Tim-3(+) T cells were evaluated. It was found burn injury induced dramatically elevated Tim-3 expression on both hepatic and splenic CD4(+) and CD8(+) T cells in contrast with the post-burn depletion of T cells. Furthermore, Tim-3 expression was correlated with the suppressive phenotype of T cells following burn injury, including increased expression of anti-inflammatory cytokine IL-10, decreased expression of pro-inflammatory cytokines IFN-γ and TNF-α, reduced T cell proliferation and elevated co-expression of Tim-3 and PD-1. Moreover, Tim-3(+) T cells subsets were more prone to spontaneous apoptosis than Tim-3(-) T cells subsets. Our findings reinforce the idea that the up-regulated expression of Tim-3 on T cells after burn injury plays an important role in the development and maintenance of burn-induced T cell immune suppression.

T cell immunoglobulin and mucin domain 3 (Tim-3) is well known to negatively regulate T cells responses,but its role in burn-induced T cells immune suppression remains unclear.In the present study,in order to identify the relationship between Tim-3 expression and post-bum T cells immune suppression,C57BL/6 mice were subjected to bum injury or sham injury,and the liver and spleen were harvested at the day 1 after operation.The expression level of Tim-3 on hepatic or splenic T cells and the functional properties of Tim-3+ T cells were evaluated.It was found burn injury induced dramatically elevated Tim-3 expression on both hepatic and splenic CD4+ and CD8+ T cells in contrast with the post-burn depletion of T cells.Furthermore,Tim-3 expression was correlated with the suppressive phenotype of T cells following burn injury,including increased expression of anti-inflammatory cytokine IL-10,decreased expression of pro-inflammatory cytokines IFN-γ and TNF-α,reduced T cell proliferation and elevated co-expression of Tim-3 and PD-1.Moreover,Tim-3+ T cells subsets were more prone to spontaneous apoptosis than Tim-3 T cells subsets.Our findings reinforce the idea that the up-regulated expression of Tim-3 on T cells after bum injury plays an important role in the development and maintenance of burn-induced T cell immune suppression.

Objective To compare the efficacy of the conventional PPD skin test and a new enzymelinked immunospot assay(TSPOT-TB)for diagnosing latent tuberculosis infection(LTBI)in patients with rheumatic diseases.Methods Two hundred and sixty rheumatic patients were enrolled,and all were screened for LTBI based on clinical history,chest X-ray,PPD skin test or TSPOT.Results The positive rate of TSPOT assay was 24.1%and that of PPD skin test was 39.4%.The overall concordance rate between the 2tests was 61.0%.Among PPD negative patients (n=149).29 were TSPOT(+)(19.5%).Among PPD(+)patients(n=98),69 were TSPOT(-)(70.0%).The patients who got BCG vaccination or had history of tuberculosis infection showed a significantly higher rate of positive result of PPD skin test than those who did not (P<0.05 or P<0.01).While in TSPOT assay,the BCG vaccination or history of tuberculosis infection did not show influence on TSPOT results(P>0.05).Of the 127 patients who received biological agents after screening for LTBI,9 patients were pretreated with isoniazide.Twenty-seven patients stopped biological agent treatment because of the positive results of PPD or TSPOT.Twenty three patients who had positive PPD but negative TSPOT results received biological agent treatment without isoniazide,and none of them developed active tubereulosis after 6 to 18 months of follow-up.Conclusion BCG vaccination affects the result of PPD test in rheumatic patients,but has no influence on TSPOT results.The infection rate of latent tuberculosis of rheumatic patients in our research is 23.8%detected by TSPOT.

Objective To evaluate a new enzyme-linked immunospot assay (TSPOT-TB) for the diagnosis of latent tuberculosis infection in patients with rheumatic diseases.Methods The rapid TSPOT-TB assay was applied to detect ESAT-6 and CFP-10 specific T cells in blood samples from 126 rheumatic disease patients.A PPD skin test was performed on all patients simultaneously.Results The positive rate of TSPOT assay was 23.8% and that of PPD skin test was 34.9%.The overall agreement between the 2 tests was 71.4%.Among PPD (-) patients (n=82),11 were TSPOT (+) ( 13.4% ).Among PPD (+) patients (n=44),25 were TSPOT(-) ( 56.8% ).The patients who got BCG vaccination showed a significantly higher rate of positive results of PPD skin test than those who did not(41% vs 19%,P＜0.05).While in TSPOT assay,the BCG vaccination did not show any influence on TSPOT results (22% vs 27%,P＞0.05).Conclusion BCG vaccina-tion affects the results of PPD test in patients with rheumatic diseases,but has no influence on TSPOT results.The infection rate of latent tuberculosis in patients with rheumatic diseases in our study is 23.8% detected by TSPOT.