Objective To observe the effects of Tongxinluo on cerebral ischemic tolerance and expression of Bcl-2 and Bax in rats with cerebral infarction.Methods The neurophysical assessment and the volume of the cerebral infarction in Tongxinluo group,middle cerebral artery occlusion(MCAO)group,preconditioning and MCAO(PC-MCAO)group were determined at different time point.The expression of Bcl-2 and Bax were measured with immunohistochemitry and the apoptosis of the nerve cells were detected with TUNEL.Results Among these groups in the corresponding time,the neurophysical marks of Tongxinluo group was the lowest and the cerebral infarction volume was the smallest at 24 h,3 d and 5 d points.The Bcl-2 positive neurons were the highest,while the Bax positive neurons and the apoptosis rate of nerve cells were the lowest around the necrotic areas in Tongxinluo group(P<0.05).Conclusion Tongxinluo can improve the cerebral ischemic tolerance and increase the expression of Bcl-2 and decrease that of Bax around the necrotic area after cerebral ischemia in rats.

ObjectiveTo determine endometrial thickness, endometrial volumeand, endometrial and subendometrial blood flows, by using transvaginal two-dimensional (2D) and three-dimensional (3D) ultrasound examination and to compare the value of all the parameters in the prediction of endometrial receptivity during in vitro fertilization-embryo transfer (IVF-ET) cycles.MethodsOne hundred and twenty patients undergoing IVF cycles were recruited for this study. Transvaginal 2D and 3D ultrasound examination was performed to determine endometrial thickness, endometrial volume, pulsatility index (PI), resisitance index (RI) , systolic/diastolic ratio (S/D) of endometrial, subendometrial blood flows , vascularization index (VI), flow index (FI) and vascularization flow index (VFI) of endometrial and subendometrial regions by experienced sonographers on the day of human chorionic gonadotropin (HCG) injection. Statistical comparison was carried out by Mann-Whitney test. The receiver operator characteristic (ROC) curve analysis was applied and the area under the curve (AUC) was calculated. Statistic analyses were performed using SPSS 13.0. The two-tailed value of was considered statistically significant.ResultsThere were no significant differences between the pregnant group and non-pregnant group in endometrial thickness, endometrial volume (P>0.05). Patients in the pregnant group had lower PI, RI and S/D of endometrial and subendometrial blood flows than those in the non-pregnant group (Z=-3.811,-3.097,-2.071, P=0.001, 0.001, 0.038), with statisticalsignificant difference. Patients in the pregnant group had higher VI, FI and VFI of endometrial and subendometrial regions than those in the non-pregnant group (Z=-1.970,-2.698,-1.981, P=0.490, 0.007, 0.048;Z=-2.098,-2.090,-2.115,P=0.036, 0.037, 0.034), with statistical significant difference. ConclusionsEndometrial and subendometria blood flows measured by transvaginal 2D and 3D ultrasound can be as a reliable indicator to assess endometrial receptivity and predict the pregnancy outcome. Among all the endometrial and subendometrial blood flow parameters, VI of endometrial and subendometrial are better predicators.

Objective To analyze the expression of purinergic receptor P2X ligand-gated ion channel 7 (P2X7R) on different cells and peripheral blood mononuclear cell (PBMC) and to investigate its correlation with inflammatory cytokines in patients with SLE.Methods Flow cytometry was used to detect surface expression of P2X7R on lymphocytes,CD4+ cells,and CD19+ cell in 29 SLE patients and 28 healthy human controls to compare the difference between the SLE patients and the controls in P2X7R expression.Enzyme linked immunosorbent assay (ELISA) was performed to detect P2X7R-related serum cytokines interleukin (IL)-1β,IL-6,tumor necrosis factor (TNF)-α level.T test,Wilcoxon rank sum test,Spearman's correlation analysis were used for statitical analysis.Results ① SLE patients had significantly higher expression of P2X7R on CD4+,CD8+ lymphocytes compared to controls [CD4+ cells∶ 2.21(3.55) vs 0.89(1.15),Z=-1.527,P=0.015； CD19+ cells∶ 11.53(20.01) vs 6.66 (6.27),Z=-2.091,P=0.037]； ② The levels of three cytokines in patients with SLE were significantly higher than those in control.The positive relationship between P2X7R expression in lymphocytes with the serum IL-6 level was found in SLE patients (r=0.449,P=0.015)；③ Patients with arthritis showed significantly higher expression of P2X7R on lym-phocytes compared to patients without arthritis (Z=-2.772,P=0.006).The expression of P2X7R on lymphocytes and CD19+ cell was significantly positively correlated with the SLEDAI score.Positive correlation with anti-β2GP Ⅰ in lymphocyteswas also found.Conclusion P2X7R may mediate the release of inflammatory cytokines involved in the pathogenesis of SLE,and may participate the development of arthritis,lupus nephritis and NPSLE in SLE patients.

Objective To study the gene expression profiles of megakaryocytes(MKs) from human cord blood CD34+ cells in vitro expanded and to understand megakaryopoiesis at the molecular level. Methods CD34+ cells were isolated using density gradient centrifugation and magnetic activated cell sorting. The cells were cultured and stimulated with recombinant human TPO ( 100 ng/ml). After 12 days, the MKs fraction was separated using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The gene expression profiles of MKs, non-MKs as well as meg-01 cells were studied by gene chip assay. THBSI, HOX A9,β-actin, lL-8,Annexin A6, FGF-8 were selected to validate the gene chip results by RT-PCR. Results A total of 116 genes between MKs and non-MKs cells were significantly different, 52 genes were up-regulated and 64 genes were down-regulated. In addition, 158 genes between MKs and meg-01 cells were significantly different, 71 genes were up-regulated and 87 genes were down-regulated. THBSI showed higher expression in MKs than in non-MKs. HOXA9 showed lower expression in MKs than in non-MKs. The expression of β-actin did not show any significant difference in MKs and non-MKs. IL-8 showed higher expression in MKs than in meg-01 cells, while ANXA6 showed lower expression in MKs than in meg-01 cells. The expression of FGF-8 did not show any significant difference between MKs and meg-01 cells. Conclusions MKs, non-MKs and meg-01 cells show different gene expression profiles. The regulatory genes include stress response genes,immune related genes, DNA synthesis and repair genes, metabolism genes, pro-onco genes and tumor suppressor genes.

Objective To evaluate the clinical applications and surgical methods of combined laparoscopic common bile duct (CBD) exploration with choledochoscopy. Methods From 2006 to 2009,clinical data of 42 patients with choledocholithiasis undergoing laparoscopic common bile duct exploration were retrospectively analyzed. We applied a step-by-step electric coagulating incision technique on the CBD,the step-by-step suturing technique, and the step-by-step clamping technique with alligator forceps, and soft tube irrigating technique with suctioning by selecting the proper exploration route, improving the common bile duct incision technique and calculus removing techniques. Results Procedures were successful in all the cases. There was no conversions to open surgery, no postoperative bleeding and no operative mortality. The mean operating time was 120 minutes (ranging, 90 to 150 minutes) with minimal intraoperative blood loss ( ranging, 20 to 40 ml). Ductal stone clearance was successful in 41 out of 42 patients ( 93% ). The largest number of the common bile duct stones was 16. With the diameter of stones larger than 15 mm in 18 cases in which the biggest was 30 mm. Bile leak developed in 1 patient, retained stones found in 3 patients,including intrahepatic cholelithiasis in one case. As a result, 38 out of 42 patients underwent common bile duct exploration. 35 patients were placed on T-tubes. Four patients underwent cystic duct exploration in which 3 had primary suture of the cystic duct and 1 had drainage. There was no infection and stenosis of biliary tract in the 42 followed-up cases. Conclusions Laparoscopic common bile duct exploration with stone extraction can be performed with high efficiency, minimal morbidity and without mortality. Improving the way of operation and selecting suitable exploration can result in better clinical outcomes.

Objective To investigate the role of endometrial and subendometrial blood flows measured by transvaginal three-dimensional(3D) power Doppler ultrasound in the prediction of endometrial receptivity during in vitro fertilization-embryo transfer (IVF-ET) cycles. Methods One hundred and twenty patients undergoing IVF-ET cycles were enrolled in this study. Transvaginal 3D power Doppler ultrasound examination was performed on the day of human chorionic gonadotropin injection to determine endometrial thickness,endometrial volume, vascularization index(VI), flow index(FI) and vascularization flow index (VFI) of endometrial and subendometrial regions by experienced sonograpbers. The patients were divided into pregnant group and non-pregnant group according to pregnant outcome. The data between the two groups were compared and analysed. Results There were no significant differences( P ＞0.05) between the pregnant group and non-pregnant group in endometrial thickness, endometrial volume. Endometrial and subendometrial VI,FI, VFI were higher in the pregnant group than those in the non-pregnant group, and statistical comparison had significant difference( P ＜0.05). Conclusions Endometrial and subendometrial blood flows measured by transvaginal 3D power Doppler ultrasound can be as a reliable indicator to assess endometrial receptivity and to predict the pregnancy outcome.

Objective To identify the immune activity of the recombinant protein Preli minarily after expressing Tir C-ter minal of enterohemorrhagic Escherichia coli(EHEC) in E.coli BL21/DE3 efficiently. Methods Tir C-ter minal (marked as Tir-C)was amplified by PCR considering the result of Bioinformatics analysis of Tir. The recombinant plasmid(designed as PET-30a(+)-Tir-C)was identified by PCR,sequencing and digested by restriction endonucleases. The positive recombinants were transformed into E.coli BL21/DE3 and induced by IPTG to express the Tir-C. The Tir-C protein was detected by SDS-PAGE and identify the antigenic of the recombinant protein Preli minarily by Western blot. Results The 675bp DNA was gained. The plasmid PET-30a (+)-Tir-C was built. Tir-C was expressed mainly in supernatant of lysis and was purified by Ni+affinity chromatograghy. Concentration of the purified protein is about 500 μg/mL and a unique band was detected with the relative molecular mass of approximately 24KDa by Western blot. Conclusion The recombinant Tir-C was expressed successfully and had immunoreactivity to some extent, which deserves the investigations for vaccine against EHEC.

Objective To evaluate the effectiveness of Percu Twist (PT) tracheostomy comparing with that of operative tracheostomy(OT)in intensive care unit(ICU). Methods Related data were retrieved from CBM,CNKI,Wanfang Data,VIP,PubMed,EMBASE,CENTRAL,and Web of Science from the time of their establishment to May 15th 2014,and the data of randomized controlled trials(RCTs)concerning PT and OT were selected. The risk of bias assessment and data extraction were performed by two independent reviewers. Meta analysis was conducted using RevMan 5.2 software. Results A total of 12 RCTs were identified,and 893 patients in ICU were involved. The results of Meta-analysis showed that PT could significantly shorten the operation time〔mean difference (MD)=-15.11,95% confidence interval(95%CI)=-17.14 to -13.07,P<0.000 01〕,reduce the volume of blood loss(MD=-17.59,95%CI=-21.90 to-13.28,P<0.000 01),reduce the size of incision(MD=-2.20, 95%CI=-2.57 to -1.82,P<0.000 01),shor ten the time of healing(MD=-3.60,95%CI=-4.15 to -3.05, P<0.000 01),and reduce complications such as infection of the wound〔odds ratio(OR)=0.20,95%CI=0.10-0.44,P<0.000 1〕and cutaneous emphysema/mediastinal emphysema(OR=0.22,95%CI=0.10-0.47,P<0.000 1)compared with OT group. The funnel plot suggested that publication bias might be found among 12 researches. Conclusions PT was shown to be more effective than OT in ICU with lower incidence of complications. As number of RCT cases is still small with unsatis factory quality,further clinical use is warranted for a better assessment.

Objective To study the therapeutic value of the endoscopic papillectomy for adenoma in the major duodenal papilla.Methods All 3 1 cases with the major duodenal papilla undergoing endoscopic papillectomy from January 2008 to June 2013 were retrospectively reviewed.The clinical data,endoscopic treatment,complications and follow-up were analysed.Results Endoscopic papillectomy was performed successfully in 29 patients.Post-operative pathology showed adenoma with low-grade dysplasia in 25 cases and high-grade dysplasia in 4 cases.One patient (3. 4%,1/29 )bled during operation;and there was no bleeding after operation. There was no perforation during and after the operation. Seventeen patients (58. 6%,17/29)developed hyperamylasemia and 3 patients(10. 3%,3/29)got mild pancreatitis,which were cured by conservative treatment.During follow-up,2 patients (6. 9%,2/29)with high-grade dysplasia had tumor recurrence after 6 months and 2 years respectively.The 27 other patients had no recurrence during the follow-up period.The shortest follow-up was 1 1 months,the longest was 5 years,with a mean time of 33. 5 ±8. 5 months.Conclusion Endoscopic papillectomy is feasible and safe for early tumors of the major duodenal papilla.Although the postoperative recurrence rate is low,it still needs close follow-up.

Objective To evaluate the effects and the safety of peroral endoscopic myotomy (POEM) for achalasia (AC).Methods POEM was performed on 7 AC patients from November 2010 to November 2011.Preoperative dysphagia grade,postoperative complications and short-term and long-term efficacy were recorded.Results POEM was performed successfully on all the 7 patients.The operation time were ranging from 46 to 113 minutes,mean 73 ± 20 minutes.Subcutaneous emphysema and mediastinal emphysema occurred in 2 patients,but cured after treatment.The 1-12 month follow-up showed dysphagia of all the patients was obviously improved.Conclusion POEM is efficient and safe to relieve dysphagia of AC patients.