Objective To establish a stable AR42J cell line expressing EGFP LC3.Methods The EGFP LC3 overexpressed Lentivirus was constructed and transfected into pancreatic acinar cells (AR42J) of rats.The rats with Lentiviral EGFP transfection were treated as negative control.The transfection efficiency was detected by inverted fluorescence microscope and flow cytometry.The EGFP LC3 protein expression in the stable cell lines were analyzed by Western blot.The cells were treated with thapsigargin to establish endoplasmic reticulum stress model,and the LC3,PERK protein expressions were detected by Western blot.Results The transfection efficiency of Lentiviral EGFP LC3 of AR42J cell was ＞ 85％,which could achieve stable passage.The expression of LC3 mRNA of AR42J cells transfected with Lentiviral EGFP LC3 was 9.14 ±0.32 folds higher than that of negative control,which had no expression of LC3 protein,only EGFP expression.However,compared with non-transfection group,the LC3 mRNA expression in EGFP group was not significantly different.Conclusions A pancreatic acinar cell line (AR42J) of rat stably expressing EGFP LC3 protein is successfully constructed.And it may provide a new model for further research of the relationship between acute pancreatitis and autophagy.

Objective To analyze the expression of interleukin (IL)-31 in tuberculous pleural effusion,and to evaluate its diagnostic value of tuberculous effusion.Methods Seventy-one patients with pleural effusion were enrolled,including 40 cases of tuberculous pleural effusion and 31 cases of malignant pleural effusion.Luminex method was applied to detect the IL-31 expression in pleural effusion.IL-31 levels were compared using non-parametric Mann-WhitneyU test,and the receiver operator characteristic (ROC)curve was used to elvaluate the diagnostic value of IL-31 .Results IL-31 expression in tuberculous pleural effusion was significantly higher than that in malignant pleural effusion with statistical significance (529.4 ng/L vs 13.8 ng/L,U =62,P <0.01 ).Based on the level of IL-31 expression,area under the ROC curve was 0.95 with the optimum cut-off value of 67.5 ng/L.Thus,the sensitivity and specificity of IL-31 ≥67.5 ng/L for diagnosis of tuberculous pleurisy were 82.5 % (95 %CI :73.3% - 94.2%)and 100.0% (95 %CI :91 .4%-100.0%),respectively.Conclusion IL-31 is highly sensitive and specific for the diagnosis of tuberculous pleural effusion, which favors the differentiation of tuberculosis from malignance.

Objective To silence the beclin1 gene expression by using RNA interference technology in AR42J rat pancreatic acinar cells,and build a stable AR42J line silencing beclin1.Methods Three kinds of shRNA targeting rat beclin1 mRNA and negative control shRNA were designed and synthesized,and were inserted into the plasmids GV112,respectively.The recombinant plasmids were named as p-sh-Beclin1-1,psh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC.Lipo3000 was used to transfect the recombinant plasmid into AR42J cells,the expression of beclin1 mRNA were detected by RT-PCR to screen for the most efficient silencing plasmid,and then it was packaged into lentiviral (LV).AR42J cells were infected with LV and screened by puromycin.Beclin1 mRNA and protein expression was determined by RT-PCR and Western blot.Results The recombinant plasmid was confirmed by agarose gel electrophoresis and sequencing showed that shRNA sequences were in line with expectations.The beclin1 mRNA inhibition rates of AR42J cells after p-sh-Beclin1-1,p-sh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC transfection were (17.8 ± 4.0) ％,(30.6 ± 2.8) ％,(45.8 ± 7.7) ％,(7.0 ± 11.8) ％,respectively.The inhibition rates of three p-sh-Beclin1 transfection cells were significantly higher than that in non-transfection cells,and the difference was statistically significant (P ＜ 0.05).While the inhibition rate of p-shRNA-NC transfection cells was not significantly different from that of non-transfection cells,p-sh-Beclin 1-3 with highest rate of inhibition was packaged by LV,and infected AR42J cells,then puromycin was applied to screen,inhibition rate of beclin mRNA expression in LV infection cells was (86.1 ± 1.2) ％,and the protein expression inhibition rate was (87.9 ± 2.8) ％,and the difference between infection and non-infection groups was statistically significant (P ＜ 0.05).Conclusions The stable AR42J line silencing beclin1 is successfully established,which can provide a new cell model for future research of the role of beclin1 in the pathogenesis of acute pancreatitis.

Objective To investigate the imbalance of peripheral Th17 cells(Th17)and regulatory T cells(Treg)in children with allergic rhinitis(AR)and bronchial asthma(BA)and its clinical significance.Methods A total of 25 children with simple AR(AR group),25 children with simple BA(BA group)and 25 children with AR complicated with BA(AR+BA group)were selected.Another 25 healthy children were selected as control group at the same time.The levels of Th17 cells and Treg cells in peripheral blood,immunoglobulin E(IgE),transforming growth factor β1(TGF-β1),percentage of forced expiratory volume in one second in predicted value(FEV1％)and levels of serum IL-6,IL-17,IL-23 and IL-10 were compared among four groups.The correlations between IgE,FEV1％and Th17,Treg,TGF-β1,IL-17 in children with AR complicated with BA were analyzed.Results There were significant differences in peripheral blood Th17 cells,Treg cells,IgE,TGF-β1 and FEV1％between AR group,BA group,AR+BA group and the control group(P<0.05).IL-17 level was significantly higher in AR+BA group than AR group and BA group(P<0.05).Correlation analysis showed that IgE was positively correlated with levels of Th17 and IL-17,and negatively correlated with Treg and TGF-β1(P<0.05).Conclusion There is imbalance of Th17 and Treg cells in children with AR and BA.Their expression are correlated with IgE and FEV1％.

Objective To investigate the imbalance of peripheral Th17 cells(Th17)and regulatory T cells(Treg)in children with allergic rhinitis(AR)and bronchial asthma(BA)and its clinical significance.Methods A total of 25 children with simple AR(AR group),25 children with simple BA(BA group)and 25 children with AR complicated with BA(AR+BA group)were selected.Another 25 healthy children were selected as control group at the same time.The levels of Th17 cells and Treg cells in peripheral blood,immunoglobulin E(IgE),transforming growth factor β1(TGF-β1),percentage of forced expiratory volume in one second in predicted value(FEV1％)and levels of serum IL-6,IL-17,IL-23 and IL-10 were compared among four groups.The correlations between IgE,FEV1％and Th17,Treg,TGF-β1,IL-17 in children with AR complicated with BA were analyzed.Results There were significant differences in peripheral blood Th17 cells,Treg cells,IgE,TGF-β1 and FEV1％between AR group,BA group,AR+BA group and the control group(P<0.05).IL-17 level was significantly higher in AR+BA group than AR group and BA group(P<0.05).Correlation analysis showed that IgE was positively correlated with levels of Th17 and IL-17,and negatively correlated with Treg and TGF-β1(P<0.05).Conclusion There is imbalance of Th17 and Treg cells in children with AR and BA.Their expression are correlated with IgE and FEV1％.

Objective To analyze the expressions of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) in the peripheral blood of patients with active tuberculosis (ATB ) or latent tuberculosis infection (LTBI) ,and to evaluate its diagnostic value in differentiation of ATB and LTBI .Methods Forty-eight patients including 18 ATB cases and 30 LTBI cases were continuously enrolled from Wuxi No . 5 People′s Hospital and Huashan Hospital affiliated to Fudan University from January 2011 to March 2013 .Flow cytometry was applied to detect the CTLA-4 expression in CD4+CD25+ FoxP3+ T cells in the peripheral blood of the 48 subjects .CTLA-4 levels were compared using non-parametric Mann-Whitney U test .Results The median percentage of CTLA-4+ Treg in CD4+ CD25+ Foxp3+ Treg cells of ATB patients was 18 .95％ (quantile range :13 .86％ ,27 .73％ ) ,and that in LTBI patients was 6 .67％(quantile range :5 .74％ ,9 .59％ ) ,which was statistically significant (U=18 .0 , P< 0 .01) .Receiver operating curve (ROC) based on the CTLA-4 expression indicated that the area under the curve was 0 .96 , with the optimum cut-off value of 13 .25％ .Thus ,the sensitivity and specificity for the diagnosis of ATB were 86 .7％ and 94 .4％ ,respectively .Conclusion CTLA-4 has highly sensitivity and specificity for the differential diagnosis of ATB and LTBI whose interferon-gamma releasing assays are all positive ,which may also provide meaningful clue for the study of pathogenesis of ATB .

Objective:To find the epidemiological characteristics of nosocomial infection in burn patients, to establish a risk prediction model for nosocomial infection in burn patients based on the screened independent risk factors of the infection, and to analyze its predictive value.Methods:A retrospective case series study was conducted. From May 2016 to December 2019, 3 475 burn patients who were admitted to the Department of Burns of Affiliated Hospital of Jiangnan University met the inclusion criteria, including 2 290 males and 1 185 females, aged from 1 to 94 years. The incidence of nosocomial infection, the detection site and specific composition of pathogenic bacteria were counted. The patients were randomly divided into training group (2 434 cases) and verification group (1 041 cases) in R 4.1.3 statistic software with a ratio of about 7∶3. Factors including gender, age, total burn area, combination of full-thickness burn/inhalation injury/shock/diabetes on admission, admission to intensive care unit (ICU), status of central venous catheterization/endotracheal intubation/urethral catheter indwelling/surgery, nosocomial infection status, days of antibiotic use, and days of hospital stay of patients were compared between the two groups. According to the occurrence of nosocomial infection, the patients were divided into nosocomial infection group (102 cases) and non-nosocomial infection group (3 373 cases), and in addition to the aforementioned data, non-nosocomial infection related data, the season of admission and types of antibiotics used were compared between the two groups. The above-mentioned data were statistically analyzed with one-way analysis of independent sample t test, chi-square test, and Mann-Whitney U test, and the indicators with statistically significant differences between nosocomial infection group and non-nosocomial infection group were included as variables in multivariate logistic regression analysis to screen independent risk factors for the development of nosocomial infection in 3 475 burn patients. On the basis of independent risk factors and important clinical characteristics, a nomogram prediction model was constructed for the risk of developing nosocomial infection of burn patients in training group. In both training group and verification group, receiver operating characteristic (ROC) curves for prediction of nosocomial infection by the prediction model were plotted, and the area under the ROC curve was calculated; calibration curves were plotted to evaluate the conformity between the predicted results of the prediction model and the actual situation; clinical decision curves were plotted to evaluate the clinical utility of the prediction model. Results:The incidence of nosocomial infection of patients included in this study was 2.94% (102/3 475); pathogens were detected from 212 specimens, mainly wound (78 cases, accounting for 36.79%) and blood (64 cases, accounting for 30.19%) specimens; 250 strains of pathogenic bacteria were detected, mainly gram-negative bacteria (153 strains, accounting for 61.20%). All clinical characteristics of patients between training group and verification group were similar ( P>0.05). There were statistically significant differences between patients in nosocomial infection group and non-nosocomial infection group in the aspects of age, total burn area, days of antibiotic use, antibiotic use type, days of hospital stay, combination of full-thickness burn, combination of inhalation injury, combination of shock, ICU admission status, central venous catheterization status, endotracheal intubation status, urethral catheter indwelling status, surgery status (with Z values of 4.41, 14.95, 15.70, 650.32, and 13.73, χ2 values of 151.09, 508.30, 771.20, 955.79, 522.67, 967.40, 732.11, and 225.35, respectively, P<0.01). ICU admission, endotracheal intubation, urethral catheter indwelling, and days of hospital stay were independent risk factors for developing nosocomial infection by 3 475 burn patients (with odds ratios of 5.99, 3.39, 9.32, and 6.21, 95% confidence intervals of 2.25-15.99, 1.56-7.39, 2.77-31.31, and 2.48-15.92, respectively, P<0.01). In training group and verification group, the area under ROC curves of the nosocomial infection prediction model based on independent risk factors, total burn area, and central vein catheterization were both 0.97 (with both 95% confidence intervals being 0.95-0.99); the calibration curve analysis showed that the prediction results of the prediction model were in good agreement with the actual situation; the clinical decision curve analysis showed that the prediction model had good clinical utility. Conclusions:The nosocomial infection in burn patients is mainly caused by gram-negative bacteria, with wound as the main infection site, and the independent risk factors including ICU admission, endotracheal intubation, urethral catheter indwelling, and days of hospital stay. Based on independent risk factors and important clinical features, the risk prediction model for nosocomial infection has a good ability to predict nosocomial infection in burn patients.

Thirty eight patients with type 2 diabetes mellitus (T2DM) signed with the Lingering Garden Subdistrict Community Health Service Center of Suzhou Gusu District from October to December 2015 were enrolled in the study. The patients were classified as low risk group (n=3), moderate risk group (n=11), high risk group (n=21) and extremely high risk group (n=3) according to risk stratification of the JADE program based on estimated glomerular filtration rate (eGFR) and risk factors [obesity, dyslipidemia, hypertension, diabetic retinopathy, urinary protein creatinine ratio, foot disease, glycosylated hemoglobin A1c(HbA1c), fasting plasma glucose(FPG) or non fasting plasma glucose]. The patients were managed by a team consisting of general practitioners, general nurses and specialists; the individualized management was implemented with reduction of controllable risk factors and complications as the goal. After one year of management (March 2016 to February 2017) the indicators of T2DM were evaluated. The results showed that the TC, LDL-C, FPG and HbA1c levels were significantly improved in both moderate and high risk groups (P<0.05); the TC, LDL-C and FPG levels in the extremely high risk group were also significantly improved (P<0.05).