ObjectiveTo examine the effects of temsirolimus, an inhibitor of mammalian target of rapamycin, on bladder cancer cell lines T24 and BIU-87 in vitro and in vivo for purpose of evaluating the probability of mTOR targeted therapy for bladder cancer.MethodsAfter being treated by a different concentration of temsirolimus, T24 and BIU-87 cells were tested by MTT assay for cell proliferation activity.Cell cycle and apoptosis analysis were performed with flow cytometer. Wound scratch assay was used for cell migration activity and transwell motility assay. Western blot analysis was used to test the mTOR phosphorylation. Subcutaneous inoculation of 6-week-old nude mice was performed using 1 × 106 T24 cells in 50％ matrigel for both control (n = 10) and temsirolimus (n = 10) groups. The volume of tumors was examined and then the expression of Ki-67 was detected by immunohistochemistry.ResultsTemsirolimus significantly inhibited proliferation of T24 and BIU-87 cells in a dose- and time-dependent manner. After administration of temsirolimus on T24 and BIU-87 cell lines for 24 h, the rate of wound healing in 0 nmol/L groups were (88.9 ± 14. 1 ) ％ and ( 83.6 ± 16.3)％ , which were higher than in the 5 nmol/L groups, which were (42.7 ± 11.6) ％ and ( 36.9 ± 9.7 ) ％ ( P ＜ 0.05 ). In the transwell motility assay, the number of cells in the 0 nmol/L group was 26.5 ± 5.8 and 28.2 ± 4.6, which was higher than in the 5 nmol/L group ( 19.0 ±3. 8 and 21.3 ± 5.1, respectively) (P ＜ 0. 05). When temsirolimus was administered on T24 and BIU-87 cell lines for 48 h the percentages of cells delayed in phase G0/G1 in 5 nmol/L group were ( 77.46 ±6.11)％ and (73. 39 ± 4. 94)％ respectively, and higher than in the 0 nmol/L group, which were (65.99 ±5.01 )％ 、(60.15 ±3.98)％ (P ＜0.05). There was no statistically significant difference in the apoptosis rate between the two groups (P ＞ 0.05 ). In Western blot analysis, the ratios of p-mTOR/β-actin were 0.92 ±0.09 and 1.01 ± 0.08 in 0 nmol/L group, and higher than in the 5 nmol/L group (0.47 ±0.05、0.04 ±0. 01 ) (P ＜ 0.05 ). After administration of temsirolimus for 21 days, the tumor volume in nude mice in the control group were 351.1 ± 139.9 mm3 , which was larger than 351.1 ± 139.9 mm3 in the temsirolimus group ( P ＜ 0.05 ). The positive rate of Ki-67 expression was ( 67.3 ± 8.4 ) ％ in the control group, which was higher than in the temsirolimus group ( 35.5 ± 6.7 ) ％ ( P ＜ 0.05 ).ConclusionsThis study provides in vitro and in vivo evidence that temsirolimus may inhibit the viability of bladder cancer cells and temsirolimus could be exploited as a potential therapeutic strategy in bladder cancer.

Objective To investigate the correlation between serum lipoprotein (a) (Lp(a)) level andischemic stroke and its etiological subtypes. Methods The consecutive inpatients with acute ischemic stroke (case group) and age-and sex-matched healthy subjects (control group) over the same period were enrolled retrospectively. The demographic and baseline clinical data, as well as fasting blood glucose, fibrinogen,homocysteine, total cholesterol, triacylglycerol, high-densitylipoprotein cholesterol, low -density lipoprotein cholesterol, and Lp(a) concentration of the case group and the control group were collected. According to TOAST classification criteria, the patients in the case group were divided into large artery atherosclerosis (LAA), small artery occlusion (SAO) and cardioembolism (CE), and the patients with other determined etiology and undetermined etiology were excluded. Multivariate logistic regression analysis was used to make clear the correlation between serum Lp(a) and acute ischemic stroke and its etiological subtypes. Results A total of 214 patients with ischemic stroke were enrolled. Ninety-seven had LAA (45.33%), 64 (29.91%) had SAO, and 53 (24.77%) had CE. There were 118 subjects in the control group. There were significant differences in the proportions of hypertension, diabetes, hyperlipidemia, atrial fibrillation and alcohol consumption, as well as systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, low -density lipoprotein cholesterol, Lp(a), fibrinogen, and homocysteine between the case group and the control group (all P <0.001). Multivariate logistic regression analysis showed that after adjustment for age and sex, Lp(a) is an independent risk factor for ischemic stroke (odds ratio [OR] 2.014, 95% confidence interval [CI ] 1.273-3.092, P = 0.036). The independent risk factors for LAA included hypertension (OR 3.353, 95% CI 1.714-6.558, P = 0.001), systolic blood pressure ( OR 2.786, 95% CI 1.136-5.538, P =0.016), homocysteine ( OR 1.108, 95% CI 1.031-2.191, P = 0.005), total cholesterol (OR 2.169, 95% CI 1.599-4.943, P = 0.001), low -density lipoprotein cholesterol ( OR2.782, 95% CI 1.093-5.238, P =0.024), and Lp(a) (OR 3.072, 95% CI 1.907-8.064, P =0.001). Theindependent risk factors for SAO included hypertension ( OR 7.042, 95% CI 3.189-25.55, P =0.001), diabetes mellitus (OR 5.162, 95% CI 2.372-11.23, P =0.001), fibrinogen (OR 1.667, 95% CI 1.434-2.025, P = 0.045), and homocysteine (OR 1.967, 95% CI 1.859-1.995, P =0.036). The independent risk factors for CE included atrial fibrillation (OR 13.340, 95% CI 4.637-39.20, P = 0.001), fibrinogen (OR 2.365, 95% CI 1.147- 4.904, P =0.029), and Lp(a) (OR 1.656, 95% CI 1.996-3.001, P = 0.035). Conclusions Lp(a) is an independent risk factor for ischemic stroke, and can be used as a serum biomarker for predicting the risk of the onset of ischemic stroke. There are differences in independent risk factors between the different stroke etiological subtypes. Lp(a) is independently associated with LAA and CE; however, it has no independent correlation with SAO.

Objective To evaluate the effect of intrathecal dexmedetomidine on the expression of G-protein-coupled inwardly rectifying K+ channel 1 (GIRK1) in dorsal root ganglia of rats with diabetic neuropathic pain (DNP).Methods A total of 144 healthy adult male SPF Sprague-Dawley rats,aged 8-10 weeks,weighing 200-220 g,were randomly divided into 4 groups (n =36 each) using a random number table:control group (group C),dexmedetomidine group (group D),group DNP,and DNP + dexmedetomidine group (group DD).DNP model was established by single intraperitoneal injection of streptozotocin (STZ) 60 mg/kg.In D and DD groups,dexmedetomidine 1.5 μg/kg was injected intrathecally at 14 days after citrate buffer or STZ injection,while the equal volume of normal saline was given in group C.The mechanical pain threshold was measured before STZ injection (T0),at 14 days after STZ injection (T1),and at 2,4 and 6 h after intrathecal injection (T:2-4).After measurement of the mechanical pain threshold at T2-4,the rats were sacrificed,and the dorsal root ganglia of the lumbar segment (L4-6) were removed for determination of the number of GIRK1 positive cells and expression of GIRK1 protein by immunofluorescence and Western blot,respectively.Results Compared with group DNP,the mechanical pain threshold was significantly increased,the number of GIRK1 positive cells in dorsal root ganglia was significantly increased,and the expression of GIRK1 was significantly up-regulated at T2-4 in group DD (P＜0.05).Compared with group D,the number of GIRK1 positive cells in dorsal root ganglia was significantly increased,and the expression of GIRK1 was significantly up-regulated at T2-4 in group DD (P＜0.05).Compared with group C,the mechanical pain threshold was significantly decreased at T1-4 in group DNP (P＜0.05).Conclusion Intrathecal dexmedetomidine attenuates DNP through up-regulating the expression of GIRK1 in dorsal root ganglia of rats.

Objective To investigate the effect of hyperbaric oxygen(HBO)combined with 5-fluorouracil (5-FU)chemotherapy on colon carcinoma Lovo cells.Methods Lovo ceils were exposed to 0.20 MPa HBO combined with 5-FU at different concentrations.The cell cycle was monitored with flow cytometry,and cell proliferation was detected using a methylthiazol tetrazolium test(MTT).Results ①After exposure to HBO,phase accumulation of Lovo cells was significantly higher than in the control group,and the accumulation of Lovo cells exposed to HBO after 24 h was significantly higher than at 12 and 48 hours.②There was difference between HBO(0.20 MPa,post 24 h)+ 5-FU(≤8 pM)group and 5-FU(≤8 μM,12 h)group on the inhibition of cells proliferation(P ＜0.05),and significan difference could be seen between the two groups after treatment for 48 h(P ＜ 0.01); ③There was no significant difference between HBO + 5-FU group and 5-FU group treatment with the concentration(16,32,64 μM)of 5-FU(P ＞ 0.05)for 12 h ;however,the difference could be seen after treatment for 48 h between the two groups(P ＜ 0.05).Conclusion The lethal effect on Lovo cells of 5-FU can be enhanced by exposure to 0.20 MPa HBO,especially with low concentrations of 5-FU.

Objective To investigate the effects of hyperbaric oxygen (HBO) on the expression of hypoxia inducible factor-1α (HIF-1α) mRNA and vascular endothelial growth factor (VEGF) mRNA in colon adenocarcinoma cells. Methods SW480 colon cancer cells were divided into a control group and an HBO group, which was in turn divided into various subgroups according to the various HBO pressures (0.15 MPa, O. 20 MPa, 0.25 MPa) and exposure times (12, 24 and 36 h) tested. The expression of HIF-1α and VEGF mRNA was evaluated with RT-PCR after the various treatments. Results The expression of HIF-1α mRNA was lower in the SW480 cells than in the controls after exposure to 0.25 MPa HBO for 12 h. No expression of HIF-1α mRNA was detected after exposure to 0.25 MPa HBO for 24 h. The expression of VEGF mRNA was lower after exposure to 0.25 MPa HBO for 36 h than in the controls. Conclusion Absence of HIF-1α mRNA expression and decreased expression of VEGF mRNA in SW480 can be observed after exposure to 0.25 MPa HBO for 24 h, suggesting that HBO may inhibit the formation of tumor blood vessels through down-regulating the expression of these mRNAs.

Objective To discuss the indication for kidney-sparing surgery (KSS) on primary urothelial carcinoma of the distal ureter.MethodsClinical data of 108 patients with primary urothelial carcinoma of the distal ureter in our hospital from 2001 to 2009 were analyzed retrospectively.There were 75 males and 33 females with mean age of 62 ( range from 42 to 85 ) years old in this study.The patients were divided into KSS group and RNU group according to the operation methods.The recurrence rate of radical nephroureterectomy (RNU) and KSS were evaluated.Results The recurrence was seen none with T,stage,1 (12.5％) with T1 stage,4 (36.4％) with T2 stage and 4 (80％) with T3 stage in KSS group.In RNU group,there was none with Ta stage,4 ( 15.4％ ) with T1 stage,10 (33.3％) with T2 stage and 7 (36.8％) with T3 stage recurred.There was no difference between patients with Ta to T2 stages in KSS and RNU group (P ＞0.05 ) on recurrence,but there was a significant difference between patients with T3 stage (P＜0.05).There was 1 (33.3％) case with G1 grade,3 (18.8％) with G2 grade and 5 (62.5％) with G3 grade recurred in KSS group,while 2 (22.2％) cases with G1 grade,9 (20％) with G2 grade and 10 (37.0％) with G3 grade recurred in RNU group.There was no difference between patients with G1 to G2 grades in KSS and RNU group (P＞0.05),but there was a significant difference between patients with G3 stage in the two groups ( P ＜ 0.05 ).Conclusion KSS seems to be safe for patients with low stage and low grade primary urothelial carcinoma of the distal ureter.

Objective To investigate the revaccination efficacy of different dosages of hepatitis B vaccine among adult non-and hyporesponders so as to improve the protection rate of hepatitis B vaccination.Methods The healthy adults who were immunized with hepatitis B vaccine at least one standard scheme in two years and negative for hepatitis B markers were enrolled in this randomized and open-label study.The hepatitis B vaccine was injected intramuscularly in the deltoid muscle on an upper arm according to routine schedule (month 0,1,6).The adults were randomly given four different dosages:73 in group A were given hepatitis B vaccine 10 μg (made in China) each time;69 in group B were given hepatitis B vaccine (made in China) 20 μg each time;70 in group C were given gene recombinant yeast hepatitis B vaccine (Engerix) 20 μg each time and 48 in group D were given gene recombinant yeast hepatitis B vaccine 40 μg each time.The serum anti-HBs was tested before and 1,2,8,12 months after the first injection.The comparison of means was done by one-factor analysis of variance and the comparison of rates was done by chi square test.Results The anti-HBs positive rates were the highest at months 8 of re-immunization in group B,C and D,which were 68.1%,70.0% and 77.1%,respectively,and were all higher than that in group A (53.4%)(χ2=21.465,P＜0.01).The anti-HBs positive rate increased in group B,C and D with increasing immunized times and over time,but there was no significant difference;it went down at 12 months after re-immunization.The anti-HBs positive rates at 1,2,8 and 12 months after re-immunization in group A were 8.2%,19.2%,53.4% and 43.8%,respectively and differences were significant (χ2=53.07,P＜0.01).The anti-HBs levels in group B,C and D were all higher than that of group A(F=7.551,P＜0.05) at month 12 of re-immunization.There were no significant differences of anti-HBs levels at different re-immunization time points in group B,C and D.Conclusion Revaecination of hepatitis B vaccine can induce immune responses and increasing dosages can improve the immune efficacy in non- and hyporesponders.

OBJECTIVE: To observe the effects of Shengqing Capsule (SQC), a compound of traditional Chinese herbal medicine, on biochemical parameters in C57BL/6J mice with cholesterol gallstone. METHODS: Thirty-eight C57BL/6J mice were randomly divided into normal control group (n=10), untreated group (n=15) and SQC group (n=13). Cholesterol gallstone was induced in mice of the latter two groups by feeding high cholesterol diet. Mice in the SQC group were intragastricly administered SQC 0.22 g/(kg.d). After 8-week treatment, animals were sacrificed and sampled to calculate the incidences of stone formation. The contents of serum cholesterols and Ca(2+), and the malonaldehyde (MDA) content and superoxide dismutase (SOD) activity in liver tissues were detected. RESULTS: The incidences of stone formation were 73.33% in untreated group, 0% in normal control group, and 23.08% in the SQC group. And the INCIDENCE in untreated group was significantly higher than those in the normal control group and the SQC group (P<0.01). Contents of serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in the SQC group were lower than that in the untreated group (P<0.01), and high-density lipoprotein cholesterol (HDL-C) in the SQC group was higher than that in the untreated group (P<0.01). SQC could increase the SOD activity and decrease the MDA content in liver tissues, showing significant differences as compared with those in the untreated group (P<0.05). CONCLUSION: SQC can decrease the incidence of stone formation and improve the biochemical parameters, which may be one of the mechanisms in the treatment and prevention of cholesterol gallstone disease.

Objective To investigate the clinical feature,diagnosis,treatment and prognosis of endometriosis of the bladder.Methods A retrospective study was conducted to review the clinical data of 10 patients with bladder endometriosis.Patient's age ranged from 30 to 48 years (with mean age of 38 years).Eight cases were admitted to hospital with urinary tract irritating symptoms during the menstrual period and 6 cases with hematuria; 2 patients without any symptoms were found through examination.The course of disease was 1-36 months (with mean of18 months).Ultrasound shows with low echo,single,wide base and no significant blood flow mass whose boundaries are less clear within the bladder wall.CT reveals soft-mass protruding into the bladder.Results Eight of the 10 patients were undergone partial cystectomy.And 2 cases was treated with transurethral resection.All cases were pathologically confirmed to be bladder endometriosis.Recurrence and ectopic lesion had not be found during follow-up period from 10 to 72 months (with mean of 30 ± 5.6 mon).Conclusions Endometriosis is a common disease in females in their reproductive years,but thebladder endometriosis is rare.The initial diagnosis needs to be made combining with imaging studies.It is confirmed by cystoscopy and pathological biopsy.Surgery is the option for the treatment of bladder endometriosis.

Objective To explore the possible protective effect of hyperbaric oxygen (HBO) on cognitive deficits induced by amyloid β25-35 (Aβ25-35) and neuronal apoptosis in the hippocampi of rats with Alzheimer's disease (AD).Methods The animal AD model was established in 24 Sprague-Dawley rats by bilateral hippocampal injection of Aβ25-35.Twelve rats were injected with normal saline as controls,and another 12 served as normal controls.After the injection,the model rats were further divided into a model group and a treatment group.All the rats were housed with normal feeding for 2 weeks and then those in the treatment groups received a total of 2 courses of HBO treatment (10 days each with an interval of 3 days in between).The other groups were left with no treatment.After the treatment,the rats' learning and memory ability were tested using Morris' water maze test,and any neuronal changes were observed using TUNEL staining.The expression of mRNA and Bcl-2 and Bax proteins in the hippocampus were detected using a RT-PCR and Western blotting.Results HBO significantly improved the learning and memory impairment and alleviated neuronal apoptosis in the hippocampus compared against the control group.In addition,HBO treatment significantly increased the mRNA and protein expression of Bcl-2 and down-regulated the expression of Bax.Conclusion HBO treatment can prevent learning and memory impairment induced by Aβ25-35 peptides,which might be mediated by inhibiting neuronal apoptosis in the hippocampus.