Objective To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression.Methods The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured.The cultured cells were treated with (1) forskolin groups: different concentration (0,2.5,5,50 or 100 μmol/L) of forskolin treated cells for 2 hours,and the optimal concentration of forskolin treated cells with different time (0,1,2,10 or 20 hours) ; (2)SP-cAMP groups: different concentration (0,2.5,5,50 or 100 μmol/L) of SP-cAMP treated cells for 2 hours,and the optimal concentration of SP-cAMP treated cells with different time (0,1,2,10 or 20 hours); (3)H-89 groups: different concentration (0,5,10,50 or 100 μmol/L) of H-89 treated cells for 2 hours,and the optimal concentration of H-89 treated cells with different time (0,1,2,10 or 20 hours).The level of intracellular cAMP and activity of PKA were detected by using ELISA,and immunohistochemistry was used to detect the localization of AQP3,the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis.Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay.Results (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group.(2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin,SP-cAMP and H-89 treatment (P ＞ 0.05).(3) After different concentration of forskolin treated 2 hours,the expression of total CREB had no significant difference among them(P ＞ 0.05).While the expression of cAMP level,PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 2.5 μmol/L,5 μmol/L,50 μmol/L forskolin group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 5 μmol/L forskolin group were higher than that in 2.5 μmol/L and 50 μmol/L (P ＜ 0.05).The optimal forskolin concentration was 5 μmol/L.(4) After different concentration of SP-cAMP treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 5 μμmol/L,50 μmol/L SP-cAMP group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 50 μmol/L SP-cAMP group were higher than that in 5 μmol/L (P ＜0.05).The optimal SP-cAMP concentration was 50 μmol/L (5) After different concentration of H-89 treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were lower in 10 μmol/L,50 μmol/L and 100 μmol/L H-89 group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 10 μmol/L H-89 group were lower than that in 50 μmol/L,100 μmol/L (P ＜ 0.05).The optimal H-89 concentration was 10 μmol/L.(6) p-CREB and AQP3 protein expression were significantly lower in 5 μmol/L forskolin combined 10 μmol/L H-89 incubating 2 hours group when compared with 5 μmol/L forskolin,but higher than that in 10 μmol/L H-89 treated group (P ＜ 0.05).Total CREB was no significant difference among the three groups (P ＞ 0.05).Conclusion cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

Aim To observe the expression of migration inhibitory factor (MIF) in the enteric neurons,and to explore the nervous regulation on MIF activity in experimental colitis.Methods Colitis was induced in sensitized rat and mouse by 2,4-dinitrochlorobenzene(DNCB)enema.MIF activity was measured both in the mesentery lymphocyte(by MTT)and in the enteric neurons(by immunofluorescence double staining).6-OHDA was intraperitonealy (ip) administered to mouse before DNCB treatment.Norepinephrine(NE) was added to lymphocyte culture in vitro during MIF preparation.Results The expression of MIF protein in enteric neuron was increased in DNCB-induced colitis in rat.ip 6-OHDA in colitis mouse(38～150 mg?kg-1) resulted in a further increase of MIF activity than ip vehicle in colitis mouse (P

Objective To evaluate the effectiveness of intern doctor training by 360 degree evaluation scale and to provide references for formulating targeted training plan. Methods A 360-degree evaluation scale with 6 subscales(Attendings, Peers, Nurse, Patients, Directors and Students) was employed to measure the status quo of medical education , represented by professional ethics and communication skills, among 129 intern doctors in medical clinical college. Cronbach's alpha was used to evaluate the reliability of the scale,one-way ANOVA was used to compare the difference of score among different scorers,and t test and Mann-Whitney test were employed to compare scores be-tween male group and female group, 5-year class and 8-year class (The significance level was set at P<0.05). Results Cronbach' α coefficients were 0.878, 0.948, 0.914, 0.908, and 0.934 for six sub-scales and 0.964 for the general scale. There were significant differences in evaluation scale scores a-mong differ-ent evaluators(P<0.01). After being adjusted by weight index, the average scores of pro-fessional ethics and communication ability & interpersonal relationship were ( 92 . 52 ± 8 . 09 ) and (93.32±8.67) respectively. No differences was observed between male and female, 5-year class and 8-year class(P>0.05). Conclusions 360 degree evaluation scale has reasonable reliability and it can provide valuable references for future similar survey in China. Meanwhile, intern doctors exhibit rela-tively good performances in professional ethics and communication ability & interpersonal relationship in the present study.

Objective To evaluate the efficacy and tolerability of zonisamide (ZNS) as add-on therapy in patients with refractory partial seizures.Methods In this Chinese muiticenter, double-blind, randomized, placebo-contrclled trial, ZNS was compared with placebo add-on therapy in 217 patients (intent-to treat (ITT) population) with uncontrolled partial-onset seizures.All patients entered a 3-month baseline period followed by a 4-week titration interval and a 12-week maintenance period.The starting dose of ZNS group was 100 mg/d, increased by 100 mg/d every week and reached the goal of 400 mg/d.The main outcome was measured by the median of the percentage of decreased seizure frequency.The secondary ouwomes points included the percentage of patients who had seizure attacks decreased by more than 50%,and adverse events.Results The median of the percentage of decreased seizure frequency in ZNS group was 33.33%, and the placebo group was 0.Thirty-eight patients (34.23%) experienced more than 50% reduction in the seizure frequency in ZNS group, compared with 19.81% of patients (21 cases) in the placebo group (χ2 =5.7159,P =0.0168) ; Moreover, 13 (11.71%) patients in ZNS group and 5 patients in placebo group were seizure free, 25 patients in ZNS group and 16 patients in placebo group who had seizure attacks decreased by more than 50%.The availability rate in ZNS group was higher than placebo group (34.23% vs 19.81%, U=2.4701, P=0.0135).The most common adverse events in ZNS group were drowsiness, fatigue, decreased appetite, gastrointestinal complaints, insomnia and constipation.Conclusion Zonisamide treatment was generally well tolerated and was associated with significant reductions in seizure frequency as adjunctive treatment for partial-onset seizures.

Objective To investigate the changes of peripheral blood expression levels of soluble fibrinogen-like protein 2 (sFGL2) in patients with chronic hepatitis B (CHB) before and after antiviral treatment.Methods From July 2013 to December 2014,initial CHB patients with entecavir antiviral therapy and healthy controls were enrolled.Clinical data at baseline and during follow-up were collected.Plasma levels of sFGL2 of all the included objects were measured by enzyme-linked immunosorbent assay (ELISA).All the patients received liver biopsy at baseline,and part of patients received a second liver biopsy at week 78 after treatment.The expression of sFGL2 in liver tissues were examined by immunohistochemistry.T test,Wilcoxon test and correlation analysis were performed for statistical analysis.Results A total of 71 CHB patients and 20 healthy controls were enrolled.At baseline,the level of plasma sFGL2 of CHB patients was significantly higher than healthy controls (105.6 μg/L (78.3 μg/L to 151.6 μg/L) vs 25.2 μg/L (18.8 μg/L to 34.3 μg/L),Z=-5.887,P＜ 0.01).The plasma sFGL2 level of patients with liver cirrhosis was 146.0 μg/L (111.3 μg/L to 166.8 μg/L),which was higher than that of patients without liver cirrhosis (79.0 μg/L (65.4 μg/L to 107.4 μg/L)),and the difference was statistically significant (Z=-4.912,P＜0.01).Plasma levels of sFGL2 were positively correlated with liver stiffness,liver inflammation and fibrosis stages (r=0.426,0.240 and 0.655;all P＜0.05).At 26 weeks and 52 weeks after treatment,the plasma levels of sFGL2 were 89.1 μg/L (69.8 μg/L to 125.5 μg/L) and 75.8 μg/L (53.4 μg/L to 98.9 μg/L),respectively,which gradually decreased compared with that at baseline (26 weeks vs baseline Z=-4.499,P＜0.01;52 weeks vs 26 weeks Z=-4.762,P＜0.01).Furthermore,at baseline the number of sFGL2 positive cells in the liver tissue of liver cirrhosis group was 33.0 ± 10.4,which was higher than that of non-liver cirrhosis group (17.6 ±6.7),and the difference was statistically significant (t=7.541,P＜0.01).Compared with that at baseline (24.5±2.0),the number of sFGL2 positive cells in liver tissue at week 78 after treatment decreased (11.3± 1.6),and the difference was statistically significant (t=11.980,P＜0.01).Conclusion Plasma level of sFGL2 is closely correlated with the degree of liver inflammation and fibrosis in CHB,and the plasma level of sFGL2 significantly decreases after long-term antiviral therapy.