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.

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.

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.

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

Objective To study the changes of renal cortex angiotensin Ⅱ (AngⅡ) expression and podocyte autophagy in aging rats with normotension versus hypertension,to further investigate the possible mechanism of renal injury in hypertension during aging.Methods Normotensive Wistar-Kyoto-rats (WKYs) were allocated to three groups by age of 3-month-old group,13-month-old group,22-month-old group.Gender,age and number-matched spontaneously hypertensive rats (SHRs) served as controls.Blood pressure was monitored.Levels of urinary albumin,urinary creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),Ang Ⅱ in serum and in renal cortex were detected.The kidney ultrastructural changes and podocyte autophagosomes were observed under light and transmission electron microscopy.Expressions of nephrin,LC3B Ⅱ,Atg5 and p62 in glomeruli were analyzed by Western blotting.Results Blood pressure was significantly higher in SHRs than in WKYs (P＜0.05).During aging from 3-month-old to 13-month-old and to 22 monthold group,the urinary albumin/creatinine ratio was increased significantly in SHRs with hypertension and in WKYs with normotension [WKY:(0.12±0.01) g/mmol,(0.14±0.04) g/mmol vs.(0.34± 0.05) g/mmol;in SHR:(0.29±0.04) g/mmol,(0.31±0.05) g/mmol vs.(0.72±0.16) g/mmol,P＜0.05],but SCR and BUN levels had no significant difference in SHRs and WKYs during aging (both P＞0.05).And Ang Ⅱ levels in both serum and renal cortex had no significant change in normotension group during aging three time points (both P＞0.05).An age-dependent decrease of Ang Ⅱ level in renal cortex appeared during aging three time points in hypertensive rats [SHR:(0.02 ±0.00) μg/L,(0.02±0.00) μg/L vs.(0.01±0.00) μg/L,P＜0.05],but serum level of Ang Ⅱ had no significant difference during aging three time points (P＞ 0.05) in hypertension group.The structural changes,including glomerulosclerosis,tubular atrophy and interstitial fibrosis were observed during aging three time points both in normotensive and hypertensive rats,but these pathological changes were more serious in hypertensive rats.Autophagosomes relatively accumulated in aging normotensive rats.The protein expressions of LC3B Ⅱ,Atg5 and p62 were increased in normotensive rats during aging (all P＜0.05).While the protein expressions of nephrin,LC3B Ⅱ,Atg5 and p62 were decreased in aging hypertensive rats (all P ＜ 0.05).Conclusions Ang Ⅱ expression level in renal cortex is decreased during aging in hypertensive rats.Podocyte autophagic activity is relatively decreased during aging in normotensive rats,but is relatively increased during aging in hypertensive rats.Ang Ⅱ-induced podocyte autophagy may be involved in the pathogenesis of renal injury in hypertension during aging.

Objective To observe the clinical effects and safety of scalp-body acupuncture and spine-manipulation therapy on preventively treating cervical cancer patients with postoperative urinary retention. Methods A total of 160 cases of cervical cancer patients with postoperative urinary retention were randomized into treatment group and control group, 80 cases in each group. Both groups were given conventional western medical treatment including preoperative indwelling of urethral catheter, clamping of urethral catheter 5 d after operation and release of the urine every 2-3 h, and trying to remove the urethral catheter 10-12 d after operation. Additionally, the treatment group received scalp acupuncture in the foot motor sensory area, body acupuncture of bilateral Shenshu (BL23) , Pangguangshu (BL28) , Ciliao (BL32) acupoints, and spine-manipulation therapy 5 d after operation. The clinical outcomes covered the incidence of urinary retention, residual urine volume, reset rate of catheter, and the clinical effect and safety in the two groups were evaluated. Results ( 1) The therapeutic effect of the treatment group was superior to that of the control group, and then the difference was significant (P<0.05). (2) After treatment, the incidence of urinary retention, residual urine volume, reset rate of catheter and average hospitalization days were less in the treatment group than those in the control group, the differences being significant ( P<0.05). ( 3) There was no adverse reaction during the treatment course. Conclusion Scalp acupuncture in the foot motor sensory area combined with body acupuncture and spine-manipulation therapy can promote the recovery of micturation function, and have satisfactory clinical effect and high safety in preventively treating cervical cancer with postoperative urinary retention.

Objective To examine the effects of benazepril and losartan on glomerular podocyte autophagy in aged spontaneously hypertensive rats (SHRs) and investigate the underlying mechanisms of renal-protective effects of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blockers (ARB).Methods Wistar-Kyoto rats (WKYs) were used as the normal control (NORM) group (2 ml physiological saline per day).SHRs were randomly divided into 4 groups:the CTRL group (2 ml physiological saline per day),the ACEI group (10 mg · kg-1 · d-1),the ARB group (30 mg· kg-1 ·d-1) and the combined group (10 mg· kg-1 · d-1 benazepril and 30 mg· kg-1 · d-1),with six 18-month-old make rats in each group.The experiments were conducted during a 4-month period.Blood pressure was monitored regularly.At the end of the experiments,we measured the levels of urine protein,urine creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),and serum and renal cortex angiotensin Ⅱ (AngⅡ).Ultrastructural changes in the kidney were examined under light and transmission electron microscopy.The expressions of nephrin,LC3BⅡ,Atg5 and p62 in the glomerulus were analyzed by Western blot analysis.Results After treatment,the blood pressure and the urine albumin/creatinine ratio of the four SHR groups were still significantly higher than those of the NORM group,but the blood pressure and the urine albumin/creatinine ratio of the ARB group and the combined group were significantly lower than those of the CTRL group (all P＜ 0.05);There were no significant differences in SCR and BUN levels among these five groups (P＞ 0.05);The level of serum AngⅡ of the combined group was significantly higher than that of the CTRL group [CTRL (0.08±0.00) μg/L,Combined (0.12±0.01) μg/L,P＜0.05];The levels of cortex AngⅡ of the four SHR groups were significantly lower than those of the NORM group,while the level of cortex AngⅡ of the ARB group was significantly higher than that of the CTRL group (all P＜0.05);Renal ultrastructural examination revealed shrunken glomeruli,fused or effaced epithelial cell foot processes,and focal atrophy of renal tubules in the four SHR groups.These pathological changes were more serious in the CTRL group but less so in the combined group.There were significantly more autophagosomes in the NORM group and the combined group than in the CTRL group (P＜0.05).Compared with the NORM group,the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the CTRL group were suppressed significantly (P ＜ 0.05).The expressions of nephrin,LC3BⅡ and Atg5 in the ACEI group and the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the ARB group and the combined group were higher than in the CTRL group (P＜0.05).Conclusions ACEI/ARB can decrease the autophagic activity of glomerular podocytes.The renal-protective effects of ACEI/ARB may be mediated by glomerular podocyte autophagy,which is induced by AngⅡ.