Objective To explore high concentrations of urea-induced human brain microvascular endothelial cell line( HBMECs) to produce inflammatory cytokines and possible mechanism .Methods HBMECs were incubated in high concentrations of urea or mannitol ( as osmotic control ) for 3,6,12 and 24 hours.Expression of TNF-αand iNOS was observed by immunofluorescence .Western blot analysis was employed to assess the protein expressions of TNF-α, iNOS, COX-2, NF-κB/P65 and p-P65.NO concentration was determined by a commercial NO assay kit . Results Immunofluorescence showed high positive immunostaining of TNF-αand iNOS after incubation in high concentration of urea stimulued as compared with control group .The protein expressions of TNF-α, COX-2 and p-P65 were significantly increased at 3 and 6 hours after high urea treatment (P<0.01), and iNOS was continued to increase from 3 to 24 hours ( P<0.01 ) .Moreover , NO content was increased at 3 hours after high urea treatment ( P<0.05 ) .Conclusions High concentration of urea can induce HBMECs to produce inflammatory cytokines .

Objective To study the effect of Tspan 8 on metastasis and invasion of human hepatocellular carcinomas(HCC).Methods RT-PCR and western blot were used to detect the expressions of Tspan 8 in HCC cell lines,HCC and matched nontumorous tissues.The expression of Tspan 8 was then down-regulated by LV/GFP/Tspan 8 in HCC cells.The expressions of Tspan 8 mRNA and protein were determined by RT-PCR and Western blot assay,respectively.The proliferation was examined by MTT,the expression of AMDM12 was assessed by Western blot,and the invasion ability of HCC cells was evaluated by transwells.Results A high level of Tspan 8 was found in high metastatic potential HCC cells,and the expression of Tspan 8 in HCC tissues was much higher than that in the matched nontumorous tissues. Down-regulation of Tspan 8 had no influence on the proliferation of HCC cells (P＞0.05),while it inhibited the expression of ADAM12 and the invasive ability of HCC cells (P＜0.01,P＜0.01 respectively).Conclusion Tspan 8 played an important role in invasion and metastasis of human hepatocellular carcinomas and down-regulation by LV/GFP/Tspan 8 inhibited the invasiveness of human hepatocellular carcinoma cells.

Objective To investigate the expression and significance of CD28,cytotoxic T-lymphocyte antigen-4 (CTLA-4),programmed death-1 (PD-1) and T cell immunoglobulin mucin-3 (Tim3) on T lymphocytes in chronic HBV-infected patients.Methods A total of 102 chronic HBV-infected patients,including 42 patients with chronic hepatitis B (CHB),30 patients with hepatitis B-induced liver cirrhosis (LC),and 30 patients with hepatocellular carcinoma (HCC),were enrolled from the First Affiliated Hospital to Soochow University during October 2012 and June 2013.Thirty healthy individuals were also enrolled as controls.Expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes in peripheral blood were determined by flow cytometry,and the differences among groups were analyzed using one-way ANOVA and LSD-t test.Spearman correlation test was performed to analyze the correlations of the expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes with HBV DNA loads,HBeAg and ALT.Results The expression of CD4 + CD28 +,CD8 + CD28 +,CD4 + CTLA-4 + in chronic HBV-infected patients were lower than those in healthy controls.CD4 + CD28 + expression in HCC group was lower than that in CHB group (t =2.373,P ＜ 0.05) ; CD8 + CD28 + expression in LC and HCC group was lower than that in CHB group (t =4.324 and 4.088,P ＜ 0.01) ; CD8 + PD-1 +,CD4 + Tim-3 + and CD8 + Tim-3 + expressions in CHB group were higher than those in LC,HCC group and healthy controls (t =3.051,3.130,3.121,3.254 and 3.723,P ＜0.01).CD8 + PD-1 + expression was positively correlated with ALT levels and HBV DNA loads (r =0.516 and 0.582,P ＜ 0.01) ; CD8 + Tim-3 + expression was also positively correlated with ALT levels andHBV DNA loads (r =0.578 and 0.556,P ＜0.01); PD-1 and Tim-3 expressions on CD8 T lymphocytes were positively correlated with each other (r =0.578,P ＜ 0.01).Conclusion The abnormal expression of the molecules on T lymphocytes in chronic HBV-infected patients is closely correlated with immune function disorder and the progression of the disease.

Objective To investigate the significance of changes in expression of co-stimulatory molecules on T lymphocytes in patients with chronic hepatitis B (CHB) infection.Methods In a casecontrol study,a total of 82 CHB cases including 50 male cases and 32 female cases (the mean age was 42.32 ±3.74) were enrolled in the First Affiliated Hospital to Soochow University from October 2012 to November 2013,together with 30 cases health control (15 male cases and 15 female cases,and the mean age was 42.32 ± 3.74).Patients were divided into three groups:40 cases of non-treated,30 cases of effective-treated and 12 cases of ineffective-treated with anti-viral drugs and immune-related therapy.The expression levels of CD28,CTLA-4,PD-1,Tim-3 on T cells subset from peripheral blood were determined by Flow Cytometry.Serum load of HBV DNA was detected by Real-time PCR and the serology markers such as HBeAg and ALT were detected by conventional methods The Kruskal-Wallis test was used to analysis groups comparison.Independent samples t-test and Mann-Whitney U test were used to two sample comparison.Spearman's rank correlation test was used to analyze the correlation.Results The expression levels of CD28,CTLA-4 and PD-1 on CD4 in health control group was the highest compared to ineffectivetreated group [404.65 (331.65-536.09) vs 277.15 (249.90-344.25) (H=29.81,P＜0.001);32.89 (29.69-39.69) vs 19.26 (11.90-20.56) (H =43.13,P ＜0.001),respectively].The expression level of PD-1 on CD8 in pre-treated group was the highest,while expression level in health control group was the lowest [15.47 (12.50-17.78) vs 3.05 (1.41-3.97) (H=56.60,P＜0.001)].Similarly,the expression level of Tim-3 on CD4 in ineffective-treated group was the highest,while Tim expression on CD4 in health control group was the lowest [199.62 (55.61-239.45) vs 70.62 (53.88-112.32) (H =41.03,P ＜ 0.001)].The expression level of Tim-3 on CD8 in pre-treated group was the highest,while it was the lowest in health control group [82.50 (78.69-84.58) vs 3.07 (1.56-6.87) (H=74.84,P ＜0.001)].Compared to the group with low viral load,the expression level of PD-1 on CD8 both in the pre-treated group with high viral load and the post-treated group was significantly increased [17.87 (13.38-20.94) (U=25.00,P＜0.001); 16.95 (5.39-18.27) (U=63.50,P＜0.001),respectively].Importantly,in the post-treated group,the PD-1 expression on CD4 T (r =0.689,P ＜0.001) and on CD8 T (r =0.751,P ＜ 0.001) was also positively correlated with ALT.Conclusions The abnormal expression of these co-stimulatory molecules may be present in the antiviral treatment process of CHB.It may provide new clues for the reasonable treatment of CHB.

Objective To investigate the predictive value of T,B,and NK lymphocyte subsets jointly detected in poor prognosis in patients with sepsis.Methods Totally 101 patients with sepsis were enrolled for prospective study.The percentages of lymphocyte sub-populations in peripheral blood were detected within 48 h after admission.The outcomes of patients during hospitalization were observed by follow-up study.The predictive value of lymphocyte sub-populations jointly detected in death risk was assessed.Results The percentages of CD3,CD3 +/CD4 + in patients with septic shock were lower than those in patients with mild/moderate sepsis and severe sepsis.The percentages of CD3+/CD8+,CD3-/CD19 +,CD4 +/CD8 + in patients with severe sepsis and septic shock were lower than those in patients with mild/moderate sepsis.The percentage of CD (16 +56) + in severe sepsis and septic shock was higher than that in mild/moderate sepsis group (P ＜ 0.05).The total number of monocytes in the deaths was higher than that in survivals.The percentages of CD3,CD3-/CD19 +,CD3 +/CD4 +,CD4 +/CD8 + were lower in deaths than those in survivals (P ＜ 0.05).The CD3-/CD19 +,CD3 +/CD8 and CD4 + / CD8 + were protective factors against sepsis with their OR values of 0.235,0.006 and 0.108,respectively.The ROC curve of lymphocyte subsets jointly detected,APACHE]] score and SOFA score to predict the risk of death were 0.993,0.877 and 0.848,respectively.The misjudgment rate of discriminating disease severity function of lymphocyte subgroups jointly detected was 19.8％,and that of outcomes was 4％.Conclusion The percentages of lymphocyte sub-populations in patients with sepsis is abnormal,and therefore,the lymphocyte subgroups jointly detected has value in the judgment of disease severity and prognosis in patients with sepsis.

ObjectiveTo investigate clinical features and outcomes in acute ischemic stroke patients with remote symptomatic intracranial hemorrhage (sICHr) after intravenous thrombolysis.MethodsThe acute ischemic stroke patients with sICHr after intravenous thrombolysis therapy were enrolled retrospectively.The clinical data were collected and the related literature was analyzed and summarized.ResultsA total of 6 acute ischemic stroke patients with sICHr were enrolled, including 4 males.Three patients had a history of using antiplatelet agents, 2 with atrial fibrillation, 4 with hypertension, 3 with previous stroke history, 4 with smoking history, and 4 had sICHr at 2 h after intravenous thrombolysis.Of the 14 hemorrhagic foci (except in the infarct areas), 10 were in the cerebral cortex.Three patients died within 1 week, and 1 was in a persistent vegetative state.Conclusions SICHr after intravenous thrombolysis in patients with acute ischemic stroke is mainly located in the cerebral cortex.The outcomes in acute ischemic stroke patients with SICHr after intravenous thrombolysis are poor, and the mortality is high.

Using the guidelines of health economic evaluation reporting is conducive to regulating the contents of this reporting, improving the quality of studies into health economics evaluation. This article summarized the history of the guidelines and specified the instrument used to study " Quality of Health Economic Studies (QHES)" and the checklist about Consolidated Health Economics Evaluation Reporting Standards (CHEERS)-the two specific evaluation contents in this guideline, the article also introduced its present application status of the guideline and its pros and cons. The checklist of CHEERS emphasized the evaluation of this report, while QHES instrument focused on quantitative evaluation on the quality of economic studies. Despite different emphasis, the two guides are actually mutually complemented.

Objective To summarize the experience in the diagnosis and treatment of hepatic trauma. Methods The clinical data of 260 patients with hepatic trauma admitted from January 1988 to December 2007 were retrospectively reviewed with regard to degree of trauma, treatment methods, therapeutical effects, complications and SO on. Results One hundred and fifty-three eases were treated by operative management,1 07 cases by nonoperative management.236 cases were cured,24 cases died,and the case fatality rate was 10.2%.There were no death among 139 patients with hepatic trauma grades Ⅰ～Ⅱ,22 death among 119 grades Ⅲ～Ⅴ patients, all death of 2 in grade VI, which demonstrated the correlation between death and hepatic trauma grade was statistically significant. Complications appeared in 82 eases, mainly including Secondary hemorrhage, abdominal infection and so on. Conclusions Nonoperative management is suitable for hepatic trauma with stable hemodynamics. Operative management is rapidly selected when the hemodynamics aren't stable. The cooperation of many specialities can enlarge the application of nonoperative management and decrease complications.

Objective:To construct an outpatient counseling record sheet of breastfeeding based on Delphi method, so as to standardize the consultation process of breastfeeding clinics.Methods:Through literature analysis and discussion of the research group, the expert letter questionnaire was designed by themselves. From September to October 2020, two rounds of Delphi letter inquiries were conducted with 16 experts in Hangzhou to determine the outpatient counseling record sheet of breastfeeding.Results:The positive coefficients of two rounds of expert letter inquiries were 100%, the expert authority coefficients were respectively 0.93 and 0.95, and the Kendall's coefficient of concordance were 0.135-0.204 and 0.234-0.563 ( P< 0.05) . The final counseling record sheet constructed included 8 first-level items, 99 second-level items and 94 third-level items. Conclusions:Letter consulting experts have high enthusiasm, high degree of authority and good coordination. The breastfeeding clinic consultation record based on the Delphi method is highly scientific and practical, which can be applied to breastfeeding clinics in various maternal and infant medical institutions.

Objective:To explore the risk factors of oral-mucosal pressure injury (OMPI) in patients with tracheal intubation in ICU and to establish a nomogram model.Methods:Using the convenient sampling method, a total of 640 patients with oral tracheal intubation admitted to ICU of the First Affiliated Hospital of Xinjiang Medical University from January to May 2023 were selected as the research objects. They were divided into the occurrence group ( n=286) and the non-occurrence group ( n=354) according to whether OMPI occurred or not. Binomial Logistic regression analysis was used to explore the risk factors for OMPI in patients with tracheal intubation in ICU. A risk nomogram model was created based on independent risk factors, and internal verification was conducted by Bootstrap repeated sampling method. Results:OMPI occurred in 286 of 640 ICU patients with tracheal intubation. Binomial Logistic regression analysis showed that high APACHEⅡ score, modified Beck oral score greater than or equal to 11 points, use of sedative drugs, prone ventilation, long retention time of tracheal catheter, low oxygenation index less than 200 mmHg (1 mmHg=0.133 kPa) and tracheal catheter fixation frequency of 1 time /24 h were the risk factors for OMPI in patients with tracheal intubation in ICU ( P<0.05). A risk nomogram model for OMPI in patients with tracheal intubation in ICU was established based on independent risk factors. The results showed that the predictive performance (area under the receiver operating characteristic curve of subjects was 0.918, 95% confidence interval was 0.897 to 0.938) and calibration (χ 2 value of 4.647, P=0.795) of the risk nomogram model for OMPI in patients with tracheal intubation in ICU were good. When the threshold probability was 0 to 1, the decision curve showed that the model had good clinical effectiveness. Conclusions:The OMPI risk nomogram model of tracheal intubation patients in ICU established in this study has good calibration and differentiation, which can be used as an effective tool for screening high-risk patients.