Aim To investigate the protective effects of sphingosine 1-phosphate ( S1 P ) postconditioning on hypoxia/reoxygenation( H/R) injury in human umbili-cal vein endothelial cells ( HUVEC ) and its mecha-nisms. Methods HUVECs cells were divided into five groups: normal ( control) group, S1P low concentra-tion group ( L ) , S1 P medium concentration group (M), S1P high concentration group ( H) and H/R group. MTT method was used to measure cell survival. Using flow cytometric analysis, the rate of cell apopto-sis was determined. The activities of total superoxide dismutase ( T-SOD) , copper/zinc superoxide dismuta-se ( CuZn-SOD ) , manganese superoxide dismutase ( Mn-SOD) activity, nitric oxide ( NO) and malondial-dehyde ( MDA ) content in cell culture medium were measured with colorimetry. Mitochondrial membrane potential in cells was observed with fluorescence micro-scope. Bax/Bcl-2, eNOS protein expression levels in HUVECs cells were observed with Western blot. Re-sults Compared with H/R group, S1P low, medium and high concentrations in the intervention group could significantly increase the cell survival rate after H/R injury, and increase activity of T-SOD, CuZn-SOD, Mn-SOD and decrease content of MDA. Moreover, S1 P could significantly increase NO content and in-crease eNOS protein expression, decrease apoptosis rate and inhibit the reduction of mitochondrial mem-brane potential. Conclusions S1P can decrease cell apoptosis rate of HUVECs after H/R injury with a cer-tain concentration dependence. The protection of S1P for cell apoptosis of HUVECs after H/R injury may be related to decreasing the intracellular MDA content and improving intracellular SOD activity, increasing mito-chondrial membrane potential and enhancing expres-sion of Bcl-2, anti-apoptotic protein.

Objective To study the therapeutic effect of EUS-guided biliary drainage (EUS-BD) on patients with malignant obstructive jaundice when ERCP failed.Methods From January 2014 to January 2016,all patients with malignant obstructive jaundice during hospitalization underwent EUS-guided biliary drainage (group A,36 cases) or PTCD treatment (group B,30 cases) by draw after failed ERCP.Operation success rate,liver function recovery time,complication rates,length of hospital stay and hospital costs were observed and compared.Results There was no significant difference in the operation success rates between two groups [94.44％ (34/36) VS 86.67％ (26/30),P＞0 05)].And there were significant differences in liver function recovery time (25.79± 6.48 d VS 30.24 ± 8.49 d),incidence of complications [5.56％ (2/36) VS 23.33％ (7/30)],length of hospital stay (21.54±4.73 d VS 25.68 ± 8.56 d) and hospitalization costs (23.5±8.4 thousand yuan VS 32.8±6.5 thousand yuan,P＜0.05).Conclusion EUS-guided biliary drainage could be the first option for its noninvasiveness and efficacy,when ERCP failed in patients with malignant obstructive jaundice.

Objective To evaluate the safety and diagnostic accuracy of preoperative endoscopic ultrasound-guid﹣ed fine-needle aspiration in patients with pancreatic cancer. Methods 256 patients with pancreatic cancer from Jan﹣uary 2010 to December 2014, 82 were considered resectable on the basis of cross-sectional imaging findings. Of these patients, 54 underwent EUS-FNA before surgery (FNA+group) and 28 underwent surgery without preoperative EUS-FNA (FNA- group), the diagnosis result of EUS-FNA and the survival time of the two groups were observed. Results All 54 lesions were visible on EUS, and all 54 attempts at FNA were technically successful. The diagnostic accuracy according to cytology and histology findings was 94.44 % (51/54) and 88.89% (48/54), respectively, and the total accuracy was 94.44 % (51/54). Two patients developed mild pancreatitis and two hemorrhage after EUS-FNA but were successfully treated by conservative therapy. No severe complications occurred after EUS-FNA. In the FNA+and FNA- groups, the median relapse-free survival (RFS) was 282 and 265 d, respectively (P>0.05), and the median overall survival (OS) was 568 and 557 d, respectively (P>0.05). RFS and OS were therefore not inferior in the FNA+group. These data indicate that the usage of EUS-FNA did not influence RFS or OS, nor did it increase the risk of other complications. Conclusions Preoperative EUS-FNA is a safe and accurate diagnostic method.

Objective To study the therapeutic effect of EUS-guided rendezvous (EUS-RV) when ERCP failed in patients with malignant obstructive jaundice. Methods 12 cases of malignant obstructive jaundice patients were underwent EUS-RV after unsuccessful ERCP. The operation success rate, liver function recovery 1 week and 1 month after operation, complication rates, hospital stay and patient survival were observed. Results All 12 patients were successfully operated and placed stents by endoscopic ultrasound puncture following by ERCP: 8 patients by the stomach, 4 patients by duodenum, the operation success rate was 100.00%; There were significant difference between the liver function recovery of preoperative and postoperative one week or postoperative one week and postoperative one month (P < 0.05). 1 bleeding occurred and were improved after conservative treatment, the complications rate was 8.33%; the hospital stay was (20.68 ± 5.76) d; the average survival time of patients was 224 d. Conclusion EUS-guided rendezvous may be the alternative treatment due to the diminutive trauma and good effect when ERCP failed in patients with malignant obstructive jaundice.

Objective To study the efficacy and safety of endobiliary radiofrequency ablation (RFA) with stent placement in treatment of unresectable extrahepatic cholangiocarcinoma (EHCC).Methods The patients with unresectable EHCC in Hangzhou First People's Hospital between October 2013 and January 2015 were enrolled in a prospective, randomized, single-blind cohort study, and were assigned randomly into two groups: RFA+stent group and stent-only group.The jaundice fade time, stent patency period, overall survival rate, and postoperative adverse events were analyzed between two groups.Results Among 59 patients with non-resectable EHCC, 28 were divided into RFA+stent group, and 31 were in stent-only group.There was no statistical difference in preoperative serum total bilirubin between the two groups.Mean serum bilirubin decrease time was significantly lower in the RFA+stent group than that in the stent-only group[17.9 d(7-22 d) VS 29.9 d(10-55 d),P=0.03].The biliary patency period in RFA+stent group was significantly longer than that of stent-only group (8.9 months VS 4.5 months, P=0.02).The mean survival time in RFA+stent group was significantly longer than that of stent-only group[13.3 months(6.2-16.5 months) VS 8.6 months(4.5-11.7 months), P=0.000).Incidence of postoperative adverse events showed no statistical difference between the two groups(P=0.727).Conclusion RFA with stent placement is effective and safe as a palliative measure in the treatment of non-resectable EHCC, and it can significantly shorten the jaundice fade time, prolong the biliary patency period and overall survival, while without increase of the incidence of adverse events.

Objective To explore the association between sleep quality and the increasing risk of type 2 diabetes mellitus (T2DM).Methods A total of 771 patients aged 25-70 years living in Xuzhou City of Jiangsu Province for at least 5 years were enrolled for the survey of risk factor related noninfectious chronic disease in 2013.In this investigation,those who suffered from other types of diabetes,neuropathy,other endocrine disease,cardiovascular,renal and hepatic dysfunction,dyspnea or cancer were excluded.To reduce the influence of confounding factors,another 771 participants were enrolled as controls.Each case was arranged to have a control who was matched in age (difference not more than 3 years),gender,residence and family history.All the participants were interviewed with self-designed questionnaire,and sleep quality was measured by Pittsburgh Sleep Quality Index (PSQI) questionnaire.Student's t test,Chi-square and multivariate logistic regression were used for data analysis.Results The PSQI score in the T2DM patients vs.the controls were 5.15±2.40 vs.2.71 ± 1.93 (t=21.96,P＜0.01).The scores of sleep-related factors,including subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency,sleep disturbance,use of sleep medication and daytime dysfunction,of the T2DM patients were higher than those of the controls.The proportion of sleep related behaviors of the T2DM patients was higher,except for early awakening,cold feeling and nightmare.Poor sleep quality was associated with the increasing risk of T2DM (odds ratio 2.06,95％ CI 1.69-2.52).In multivariate logistic regression,when adjusted for confounding factors,the risk of T2DM was still increased (odds ratio 1.72,95％ CI 1.62-1.83).Sleep-related factors (e.g.subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency and sleep disturbance) were correlated with the risk of T2DM (odds ratio was 3.34,1.63,1.10,1.87 and 3.89,respectively).Conclusion Low quality of sleep may be strongly associated with an increased risk of T2DM.

Objective To evaluate the relationship between ultrasound-assayed detrusor thickness and bladder outlet obstruction (BOO) in old men with benign prostatic hyperplasia (BPH).Methods The 106 BPH patients underwent the urodynamic examination on which the diagnosis of BOO was dependent. The obstruction was defined as the Abrams-Griffiths nomogram (A-G index)≥40 and the grade of linear passive urethral resistance relation (LinPURR)≥Ⅱ . When bladder capacity reached 150 ml, the detrusor thickness was measured by abdominal ultrasound. Results Compared with unobstructed group, the maximum flow rate and average flow rate were both lower in obstructed group [(10.1±3.0) ml/s vs. (17.4±3.1) ml/s, (5.5±2.2) ml/s vs. (11.2±2.2) ml/s, t= 10.26and 11.03, both P＜0.01]. And the residual urine volume and maximum detrusor pressure were significantly higher in obstructed group than in unobstructed group [(47.6 ± 24.3) ml vs. (17.0 ±5.6) ml, (39.3±14.4) cm H2Ovs. (26.8±8.0) cm H2O, t=6.32 and 4.07, P＜0.01 or 0.05].Detrusor thickness was positively correlated with maximum detrusor pressure (r= 0.419, P＜0.01),but negatively correlated with maximum flow rate (r =- 0.749, P＜ 0.01 ), mean flow rate (r=-0.853, P＜0.01) and voided volume (r=-0.556, P＜0.01). There was significant difference in detrusor thickness between obstructed group and unobstructed group [(3.0± 0.2) mm vs. (2.5 ±0.2) mm, t= 11.2,P＜0.05]. According to the diagnostic standard of detrusor thickness≥3.0 mm,it had a sensitivity of 90% and a specificity of 84.6%, a positive predictive value of 93.1% and a negative predictive value of 78.6%. Conclusions Detrusor thickness of 3.0 mm or greater has a certain predictive value for BOO in old men.

Objective To summarize the effects of medical treatment of urinary calculi in 112 infants under 3 years old with a history of feeding melamine-contaminated milk powder.Methods All the infants with a history of feeding melamine-contaminated milk powder[73 male,39 females;aged(14.43?8.63) months] were diagnosed with type-B ultrasonic,and through the main treatment procedures were 50 g/L sodium bicarbonate(0.5 mL/kg,intrarenous injection) and fluid infusion.The dosage of drugs were timely adjusted according to the monitored uine pH.Hemodialysis was performed in the patients with renal failure.These were decided on the basis of the effect of medical treatment.Results During their hospitalization of 3 to 25 days,56 cases obtained full response,49 cases showed partial response and 7 cases demonstrated no response.The mean diameters of calculi were(4.77?3.16) mm,(8.13?3.79) mm and(6.92?2.65) mm,and the average pH values were 5.64?0.11,5.75?0.10 and 6.38?0.31 in full response,partial response and no response groups,respectively.The average pH value in no response group was significantly higher than that in full response group(P0.05).Conclusions Medical treatment can help to obtain considerable therapeutic effects for infants under 3 years old with urinary melamine calculi.Treatment response has shown partial relationship with the size of urinary calculi and pre-treatment urine pH,while no relationship is found to be related to infants′ age.

Objective To study the effects of fluoride on minichromosone maintenance(MCM)3 mRNA and the bone formation-related gene:bone sialoprotein(BSP),osteocalcin(OC),osteopontin(OP)mRNA expression on human osteoblast cells.The expression of MCM3 was tested for diagnosis and surveillance value on osteoblast treated with excess fluoride.Methods Human osteoblast cell(Saos-2)was cultured in McCoy5A medium and treated with fluoride(sodium fluoride,NaF).There were eight groups including:0(control),0.625,1.250,2.500,5.000,10.000,20.000,40.000 mg/L groups.Expression of MCM3,BSP,OC,OP mRNA were detected by real-time PCR.Dual-standard curve method was used for analysis.ALPase was determined by measuring the absorbance using a micro titer plate reader. Results Expression of MCM3 mRNA was lower in the 0.625,1.250,2.500,5.000,20.000, 40.000 mg/L groups(0.059 ± 0.003,0.027 ± 0.001,0.272 ± 0.004,0.115 ± 0.002,0.137 ± 0.004,0.754 ±0.002, all P > 0.05) and was higher in10.000 mg/L group(21.300 ± 1.200, P < 0.01 ) than control group( 1.000 ±0.020), especially 10.000 mg/L group was higher than groups treated with fluoride(all P < 0.01 ), the differences among groups were significant(F = 305.842, P < 0.01 ). Expression of BSP mRNA was significantly higher in 0.625,1.250,2.500,5.000,10.000 mg/L groups(71.80 ± 3.60,133.00 ± 7.20,85.50 ± 0.60,80.90 ± 1.20,304.00 ± 21.00)than the control group( 1.00 ± 0.04), especially 10.000 mg/L group was higher than others groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 159.531, P < 0.01 ). Expressions of OC mRNA were higher in 0.625,1.250,2.500,5.000 mg/L groups(110.00 ± 12.00,143.00 ± 2.10,90.60 ± 4.10,23.70±1.20) than control group(1.00 ± 0.01, all P < 0.01), and the differences among groups were significant (F = 158.734, P < 0.01 ). Expression of OP mRNA were higher in 0.625,1.250,2.500,5.000,10.000,20.000 mg/L groups(167.00 ± 11.20, 111.00 ± 12.10,72.50 ± 3.50,134.00 ± 14.00,42.30 ± 2.40,45.20 ± 3.30) than the control group(1.00 ± 0.04, all P < 0.05 or < 0.01 ), the differences among groups were significant(F = 60.226, P < 0.01 ).Compared with control group(4.2 ± 1.2), the ALPase activity was increased in all groups treated with fluoride (6.0 ± 0.4,5.8 ± 0.1,5.7 ± 0.4,7.7 ± 1.1,19.2 ± 2.4,8.5 ± 3.0,18.1 ± 4.2), but only 10.000 mg/L and 40.000 mg/L groups were higher than control group and other groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 7.806, P < 0.01 ). Conclusions Irregular expression of MCM3 mRNA is not suitable as a diagnostic and monitoring biomarker of osteoblasts exposed to excessive fluoride. Fluoride may affect the osteoblast-related gene expression and to promote osteogenic differentiation.

Objective To observe the lecithin's effect on membrane of African green monkey kidney cells (Vero) exposed to sodium arsenite(NaAsO2). Methods Vero cells cultured in vitro were divided into 4 groups:control group (saline), model group (2.20 mg/L NaAsO2), high eoncentration of lecithin and arsenic group (53.33mg/L lecithin + 2.20 mg/L NaAsO2), low eoncentration of lecithin and arsenic group( 13.32 mg/L lecithin + 2.20 mg/L NaAsO2), 6 bottles of cells in each group, medium was changed every 2 days, cultured for 120 h. Na+ ,K+-ATPase activities of membrane were measured by spectrophotometry, and membrane phospholipids composition including phosphatidylserine (PS), phosphatidylethano-lamine (PE), phosphatidylcholine (PC) and sphingmyelin (SM) were measured by high performance liquid chromatography (HPLC). Results The Na~, K+-ATPase activities of membrane of control group, model group, high concentration of lecithin and arsenic group, low concentration of lecithin and arsenic group were (0.962 ± 0.081) × 106, (0.544 ± 0.037) × 106, (0.647 ± 0.043) x 106, (0.550±Compared with control group, the Na+ ,K+-ATPase activities of other 3 groups were significantly reduced (all P < 0.05). Compared with model group, the Na+ ,K+-ATPase activity in high concentration of lecithin and arsenic group was significantly higher (P < 0.05),but in low concentration of lecithin and arsenic group did not change significantly (P > 0.05). Compared with control group[(0.087 ± 0.003), (0.127 ± 0.053), (0.588 ± 0.105),(0.071 ± 0.029)g/L], PS, PE, PC, SM levels in model group[(0.051 ± 0.018), (0.073 + 0.030), (0.240 ±0.038), (0.047 ± 0.121 )g/L] were significantly lower(all P < 0.05) ;PS, PE, PC in high concentration of lecithin and arsenic group[(0.084 ± 0.011), (0.109 ± 0.363), (0.591 ± 0.476)g/L] did not change significantly(all P > 0.05), but SM[(0.057 ± 0.004)g/L] significantly decreased(P < 0.05) ;PS, PE, SM levels of low concentration of lecithin and arsenic group[(0.058 ± 0.020), (0.086 ± 0.177), (0.048 ± 0.103)g/L] significantly reduced (all P < 0.05), the PC did not change significantly [(0.521±0.098 )g/L, P > 0.05]. Compared with model group,the levels of PS, PE, PC, SM in high concentration of lecithin and arsenic group were significantly higher(all P <0.05);PS, PE, SM levels in low concentration of lecithin and arsenic group did not change significantly(all P > 0.05), and PC was significantly higher(P < 0.05). Conclusions High concentration lecithin has certain protective effect on Vero cell membrane exposured to sodium arsenite.