Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) or high glucose on the toll-like receptor 4 (TLR4) expression,inflammatory cytokines and fibrotic factors in human tubular epithelial cells (HK-2),revealing the innate immune-related pathogenesis of diabetic nephropathy (DN) which may have clinical implications.Methods Three TLR4 siRNA sequences were designed and synthetized.After transfection,the most effective siRNA was selected to use for further expriments.The experiment consisted of 2 parts.Part 1:Cells were divided into three groups:normal-glucose group (NG,5.5mmol/L glucose),mannose group (M,5.5 mmol/L glucose + 19.5 mmol/L mannose),High-glucose group (HG,25 mmol/L glucose),preliminary validated the effects of high glucose and high osmotic pressure.Part 2:Cells were divided into seven groups:NG group,HG group,Ang Ⅱ group,Ang Ⅱ + negative group,HG+ negative group,Ang Ⅱ + siRNA group and HG+ siRNA group.Real time PCR was used to analyze the mRNA expression of TLR4,myeloid differentiation factor 88 (MyD88),heat shock protein 47 (HSP47).Western blotting was used to observe the protein expression of TLR4,MyD88,HSP47,NF-κB,type Ⅳ collagen (ColⅣ).ELISA was used to detect the expression of monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6).Results Compared with NG group,TLR4,MyD88,HSP47 mRNA and TLR4,MyD88,NF-κB,ColⅣ,HSP47 protein were highly expressed under high glucose or Ang Ⅱconditions (P ＜ 0.01),and the expression levels of MCP-1 and IL-6 also increased significantly (P ＜ 0.01).Compared with HG or Ang Ⅱ group,the above indicators were obviously inhibited in the TLR4 siRNA groups (P＜0.01).Comparison between blank vector transfected groups and HG group as well as Ang Ⅱ group indicated no statistic significance (P ＞ 0.05).Conclusions Both Ang Ⅱ and high glucose stimulate TLR4 expression,which result in the up-regulation of inflammatory and fibrotic factors in HK-2.Specific target of TLR4 gene silencing can block the TLR4 pathway that is activated by high glucose and Ang Ⅱ,and thus reduce the inflammatory and fibtogenic factors' release.TLR4 signal is the common innate immune response pathway which induces the release of inflammatory and fibrotic factors in HK-2 under high glucose or high angiotension conditions.

Objective To evaluate the efficacy and safety of InVance bulbourethral sling for stress urinary incontinence after radical prostatectomy.Methods Six male patients with stress urinary incontinence after radical prostatectomy were treated with perineal sling (InVance) from March 2005 to March 2011,and the clinical data were analyzed.Results The operation time was 80-120 min and intra-operative blood loss was 20-50 ml.The catheter was removed at the second day after surgery.In a follow-up of 12 to 29 months,4 patients were dry,1 patient was improved,and 1 patient suffered treatment failure.There were 2 patients transient acute urinary retention and 1 patient short-term perineal pain in the main post-operative complication.In six months after the operation,urodynamic examination showed (15.9 ± 7.7) ml/s at Qmax and 0-20 ml/s at postvoid residual.Conclusion InVance bulbourethral sling for the treatment of stress urinary incontinence after radical prostatectomy appears to be safe,simple in the procedure and good effect to mild and middle incontinence.

In medical clinical study, the researchers and the ethics committee members' personal economic in-terests and responsibilities conflicts may produce damage to experimental research and the subjects, may make the study being questioned the authenticity and the objectivity of clinical trial results, also damage to the subjects' rights and interests and damage the credibility of hospital. In order to prevent the happening of the conflict, put forward the strategies:strengthen legislation construction, improve the treatment method, set up a conflict of interest com-mittee, adhere to the principle of open, review and restriction to the personnel, regularly organize researchers and ethics committee members attend the training.

This study was aimed to develop an HPLC method for the determination of hesperidin,magnolol,honoki-oland liquiritin in Soft Capsule Jia-Wei Huo-Xiang Zheng-Qi (JWHXZQ). AKromasil C18 column (250 mmí4.6 mm, 5 μm) was used with water-methanol as mobile phasegradient elution. The flow rate was 1.0 mL·min-1, and the de-tecting wavelength was at 287 nm. The results showed that the linearity ranges ofhesperidin,magnolol,honokioland liquiritinwere 4.47-178.70 μg·mL-1, 3.42-136.96 μg·mL-1, 3.49-139.48 μg·mL-1, 3.92-157.20 μg·mL-1, respec-tively (r>0.999). The average recoveries of them were 99.48%, 99.05%, 99.57% and 99.79%, respectively. It was concluded that the method was accurate, sensitive and specific for quality control of Soft Capsule JWHXZQ.

Objective To investigate the relationship of pancreatic β-cell function and insulin resistance with microalbuminuria in a cross -sectional study of patients with type 2 diabetes.Methods A total of 524 partici-pants with type 2 diabetes were recruited in this cross -sectional study.All subjects'height,weight,waist circumfer-ence and blood pressure were measured.Venous blood samples were drawn to measure fasting plasma glucose (FPG), fasting lipids,glycated hemoglobin A1c (HbA1c),fasting C -peptide (FPC).24h -urine was collected to measure urinary albumin excretion rate (UAER).Homeostasis model assessment of pancreatic β-cell function (HOMA -B) and insulin resistance (HOMA -IR)were estimated using fasting plasma C -peptide.According to HOMA -B quar-tile,the subjects were divided into four groups,including q1 -q4.According to HOMA -IR,the subjects were also divided into four groups,including Q1 -Q4.We assessed the crude associations across quartiles of these data with demographic and clinical parameters using a nonparametric test for trend across ordered groups (trend using Stata software).Multivariable logistic regression analysis was performed to assess the relationships of pancreatic β-cell function and insulin resistance with microalbuminuria in patients with type 2 diabetes.Results Trend test showed that UAER gradually reduced with increase of HOMA -B.The UAER values in subjects with q1,q2,q3 and q4 were 8.92(5.53 -28.65),8.55(5.52 -20.95),7.57(4.79 -19.83)and 7.84(5.23 -14.38)μg/min,respectively, and the trend was statistically significant(z =-2.1,P <0.05 ).With HOMA -IR increasing,UAER gradually increased.The UAER values in subjects with Q1,Q2,Q3 and Q4 were 6.73(4.85 -16.52),8.61 (5.2 -20.37), 8.31(4.88 -27.04),8.75(6.03 -25.21)μg/min,respectively,and the trend was also statistically significant(z =2.41,P <0.05).Multivariable logistic regression analysis showed that subjects with the highest quartile of HOMA -B had lower possibility of microalbuminuria than patients with the lowest quartile of HOMA -B (adjusted OR q4 vs. q1 =0.39,95% CI:0.20 -0.76,Wald =7.59,P =0.006).Subjects with the highest quartile of HOMA -IR had higher risk of microalbuminuria than those with the lowest quartile of HOMA -IR (adjusted OR Q4 vs.Q1 =2.00, 95% CI:1.08 -3.72,Wald =4.84,P =0.028).Conclusion Insulin resistance is associated with an increased prevalence of microalbuminuria in type 2 diabetes,while improved pancreatic β-cell function is linked to decreased rates of microalbuminuria for those patients.

OBJECTIVETo construct and characterize the TGF-β1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction.

METHODSThe epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-β1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant.

RESULTSThe mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-β1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-β1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05).

CONCLUSIONSTGF-β1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-β1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.

Biomarkers ; metabolism ; Cadherins ; genetics ; metabolism ; Epithelial Cells ; drug effects ; physiology ; Epithelial-Mesenchymal Transition ; drug effects ; physiology ; Humans ; In Vitro Techniques ; Keloid ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; pharmacology ; Up-Regulation ; Vimentin ; genetics ; metabolism

OBJECTIVETo investigate the result of hip arthroplasty for failed internal fixation of femoral neck fractures.

METHODSFrom June 2007 to January 2014, 29 cases who underwent hip arthroplasty for failed of internal fixation of femoral neck fractures were reviewed. There were 12 males and 17 females. The mean age was 60.3 years (ranged 43 to 83 years) at the time of the fracture. Left hip was in 16 cases, right hip was in 13 cases. The average interval from fracture to arthroplasty was 23.3 months (ranged, 3 to 48 months).

RESULTSAll of 29 cases were performed total hip arthroplasty. There were 20 cases of cementless cup,7 cases of cementless cup with bone graft, 2 cases of cemented cup with bone graft; 13 cases of cementless stem, 16 cases of cemented stem. There were no complications occurred such as intraoperative fracture of the greater trochanter. The average operative time was (115 ± 38) minutes,the mean intraoperative blood loss was (420 ± 175) ml, the average postoperative drainage volume (240 ± 119) ml, intraoperative blood transfusion was (200 ± 220) ml, intraoperative fluid volume was (2,200 ± 400) ml, the average postoperative blood transfusion was (300 ± 200) ml. There was 1 case get postoperative dislocation. All patients were followed up for 14.7 months in average (ranged, 5 to 24 months). There was no revision for mechanical failure. Harris Hip Score significantly was improved from 51.1 ± 7.5 before the conversion to 88.5 ± 6.4 points at the final follow-up.

CONCLUSIONThe effect of the hip replacement for patients with failed internal fixation of femoral neck fractures was confirmed. This method can shorten the time on the bed and reduce the complications. It benefits the patients earlier functional recovery, but it must control operation indication. The long term efficacy is necessary to further observation.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Treatment Failure

This study was aimed to screen main active site to reduce blood lipid from Xin-Mai (XM) capsule and establish HPLC fingerprint of the site, in order to study the correlativity between active site and relevant fractions of its herbs. Solvent extraction was used to separate XM capsule into different polar fractions. Intraperitoneal injection of 75% egg-yolk emulsion was used to establish mice hyperlipidemia models. And the active site was screened. Chromatographic fingerprints of the site and relevant fractions of its herbs were configured by HPLC analysis. The
retention time of peaks was utilized as index to evaluate the correlativity. The results showed that lipid-lowering effect of ethyl acetate extract and garlic essential oil was significant (P<0.01). Fingerprint of the active site in XM capsule was established with 28 fingerprint peaks and the assignment results of 27 peaks were indicated. It was concluded that the active sites to reduce blood lipid of XM capsule were ethyl acetate extract and garlic essential oil. The established fingerprint method can effectively determine the correlativity between the active site and its relevant fractions, which contributed to pharmacodynamic material foundation and quality standard.

Objective To evaluate efficacy and safety of ursodeoxycholic acid(UDCA)for preven-tion of obstruction of biliary plastic stent.Methods A total of 88 patients with plastic biliary stents at our hospital were divided into UDCA group and the control group.Stents were collected 3 months after the opera-tion for argile biliare in the stent.The contents of APF,calcium bilirubin and calcium carbonate were ana-lyzed by biochemistry,and bacterium culture was conducted.Results The obstruction rate of UDCA group (n=43)and control group(n=45)were 8. 9% and 58. 1% respectively with significant differences(P<0. 05).There were significant differences in the protein level of APF between the UDCA group(17. 29 ± 9. 52)μm/L and the control group(10. 39 ±2. 17)μm/L(P<0. 05).Calcium bilirubin[(13. 90 ±3. 80) vs.(30. 92 ±7. 07)]and calcium carbonate[(12. 60 ±5. 69)vs.(16. 52 ±4. 11)]were less than those of the control group (P<0. 05 ).Only one type of bacterium was found,and Escherichia coli were cultured in 40. 0% samples,a lower incidence compared with the control(72. 1%).Conclusion UDCA would lessen biofilm and smooth stent surface by promoting secretion of endogenous bile acid and increasing APF,and keep the biliary patency.

OBJECTIVE Toexplorethefeasibilityofpreparingamigrainemodelusingratdura materofsuperiorsagittalsinusinfollowingelectricalstimulation.METHODS MaleSDratswereexposed to 1 h electrical stimulation after brain electrode implantation,at 3 V,6 Hz,and a pulse of 0.25 ms.At 0, 1 5,30 min,and 1 h,orbital blood and brain tissue were taken.The content of calcitonin gene-related peptide (CGRP),substance P and prostaglandin E2 (PGE2 )in the plas ma and brain tissue was detec-tedbyradioimmunoassay.RESULTS ThebraintissuecontentofCGRP,substancePandPGE2in the sham group and normal group was not significantly different.CGRP,substance P and PGE2 in brain tissue of model group were 3.41 ±0.93,1 .80 ±0.64,3.41 ±0.93 and (1 .80 ±0.64)ng·g -1 respec-tively,significantly different from sham group(P<0.05).At different time points,the content of CGRP, substance P and PGE2 in plasma orbital venous plexus showed no significant difference between sham groupandmodelgroup.CONCLUSION ThereleaseofCGRP,PGE2andsubstancePinbraintissue is significantly different fro m that of plas ma after electrical sti mulation.The results suggest that a model of migraine can be constructed by electrical stimulation.