Objective To discuss the curative effects of alfacalcidol combined with losartan potassium tablets in the treatment of early diabetic nephropathy (EDN).Methods 90 EDN patients in our hospital were chosen and randomly divided into the observation group and control group (45 cases in each group).The control group was given losartan potassium tablets treatment,while the observation group was given alfacalcidol combiend with telmisartan treatment.All the two groups were treated for 3 months.Before and after treatment,the fasting blood glucose (FBG), 24h urine trace albumin quantitative (UAER),24h urine protein (24h pro),serum creatinine (SCr),25 -hydroxyl vitamin D [2 5 (OH )D ],blood calcium (Ca2+),potassium (K+),glycosylated hemoglobin (HbA 1 C )and serum inflammatory factors[C-reactive protein (CRP),tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6 )] were observed,and the correlation between 25 (OH)D and UAER,24h pro was analyzed.At the same time,the clinical curative effects and adverse reactions during treatment were evaluated.Results In the observation group,the total effective rate was 93.3%,which was significantly higher than 71.1% in the control group (χ2 =7.601,P<0.05).In the two groups after treatment,the FBG,HbA1c,blood Ca2+and K+had no significant changes (all P>0.05).After treatment,Scr,24h pro and UAER in the two groups were all significantly reduced compared with before treatment (P<0.05),and 24h pro and UAER in the observation group were significantly lower than those in the control group (t=6.296,11.530,all P<0.05),but Scr had no statistically significant difference between two groups (t=0.331,P>0.331).After treatment,25(OH)D in the observation group decreased significantly compared with before treatment (t=12.000,P<0.05),which was significantly higher than that in the control group after treatment (t=11.278,P<0.05).Compared with before treatment,25(OH)D in the control group had no significant change after treatment (t=0.436,P>0.436).Pearson correlation analysis showed that 25 (OH)D level was negatively correlated with 24h pro and UAER (r=0.483,0.778,all P<0.05 ).In the two groups after treatment,the CRP, TNF-αand IL-6 levels significantly reduced (P<0.05 ),which in the observation group decreased more obviously (all P<0.05).In the two groups,no serious adverse events were found,the difference was not statistically significant (χ2 =0.212,P >0.151 ).Conclusion Alfacalcidol combined with losartan potassium tablets can significantly reduce the proteinuria levels of EDN patients and inflammation,which has better clinical curative effects and higher safety.

Objective To study the stability of micro-implant orthodontic anchorage(MA)with different pitch in the case of immediate loading.Method Employing 3D finite element analysis method,the stress and dis-placement distribution on the bone interface of MIA with different pitch(0.3 mm、0.5 mm、0.7 mm and 1.0 mm,respectively),which was 1.47 N loaded vertically in the major axis direction,were analyzed.Result The pitch affected the stress distribution significantly,because the maximum stress increased with the pitch decreasing and the impact of pitch on stress distribution on neck and central locations of MIA were different;to decrease the pitch could reduce the max displacement of the jaw,but the impact of pitch on displacement distribution of MIA was not significant.Condusions In the case of immediate loading.MIA with pitch 0.5 mm-0.7 mm is suggested to be selected as orthodontic anchorage in the clinic.

Objective To construct the recombinant plasmid of PA-ⅠL and express in E.coli BL 21 (DE3), and evaluate the biological activity of recombinant protein.Methods PA-ⅠL gene was amplified by PCR using primers designed according to Pseudomonas aeruginosa genome sequences and then cloned to the vector pET -28a ( +).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced to express by IPTG.The recombinant protein was purified by nickel affinity chromatography.The binding activity of recombinant PA-ⅠL with Gb3/CD77 was evaluated by flow cytometry.The function of recombinant PA-ⅠL on the binding of bacteria with host cells was evaluated by colony plate counting.Results and Conclusion The recombinant PA-ⅠL protein was highly expressed in E.coli BL21(DE3) and protein purity by SDS-PAGE analysis was high after nickel affinity chromatography .Besides, the recombinant PA-ⅠL had binding activity to Gb3/CD77 and inhibited the binding of PAO1 to host cells in a dose-dependent manner.

Objective:To construct the eukaryotic expressed vector which express recombinant toxin CD80 Linker SEA and predict the rationality and feasibility of the linker Methods:Utilize the sequence analysis software to analyze the flexibility、antigenicity、Hoop&Woods hydrophilicity and episode of recombinant toxin CD80 Linker SEA Results:Through the analysis of the software,it could be found that the recombinant toxin has correct domains of CD80 and SEA The linker has low episode、low antigenicity and high flexibility Conclusion:The results of computer analysis could help us to rationally design the recombinant toxin CD80 Linker SEA and keep it's maximum biological activity

Objective To investigate the association between CYP11 B2 gene polymo-rphism and left ventricle hypertrophy with meta analysis.Methods Literatures about the association of CYP11 B2 gene polymorphism and left ventricle hypertrophy from January 1992 to December 2011 were searched.The electronic databases retrieved from Pubmed,Embase,China national knowledge intemet,Chinese biological medicine disk,VIP fulltext database and Wanfang fulltext database.Odds ratio of CYP11 B2 genotype distributions in left ventricle hypertrophy patients comparing with healthy control were analyzed.RevMan5.1 software was applied for investigating hereogeneity among individual studies and summarizing effects with proper statistical methods.Six case control studies were enrolled.Results A total of 541 cases and 553 controls were enrolled for the study.The pooled OR of CC vs TT + TC genotype was 1.15 (95％ CI:0.74 ～ 1.80) (Z =0.63,P =0.53) in the subgroup of hypertension,and the pooled OR of CC vs TT + TC genotype was 1.15 (95 ％ CI:0.74 ～ 1.80) (Z =0.63,P =0.53) in the subgroup of race.The pooled OR of C vs T allele was 1.15 (95％ CI:0.76 ～ 1.74) vs 0.87 (95％ CI:0.58 ～ 1.31) (Z =0.67,P =O.50).Conclusion Whether the hypertension or the race,the genotype of CYP11 B2 polymorphism has no association with an increased risk of left ventricle hypertrophy.

Objective To analysis of risk factors for left atrial thrombosis in patients with rheumatic mitral stenosis.Methods From January 2001 to December 2008, 2277 patients with rheumatic mitral stenosis underwent operations in our hospital. There were 737 males and 1540 female, the age ranged from 19 to 84 years [average (50.9 ±10.2) years]. Left atrial thrombosis group (554 cases) and no thrombosis group (1723 cases) were divided, retrospectively collected data were analyzed with univariate and multivariate Logistic regression. Results 12 bvariables, including age, mitral valve orifice area, left atrial diameter, left ventricular diastole diameter, CRP, gender , degree of mitral stenosis, or regurgitation, degree of bicuspid regurgitation, degree of pulmonary hypertension, atrial fibrillation and heart function had statistic difference between two groups. With multivariate Logistic regression for these 12 factors, age, mitral valve orifice area, left atrial diameter, degree of mitral regurgitation and atrial fibrillation were found to be the affecting factors for left atrial thrombosis in patients with rheumatic mitral stenosis. Conclusion For patients with rheumatic mitral stenosis, age, mitral valve orifice area, left atrial diameter and atrial fibrillation are the risk factors for left atrial thrombosis. Mitral regurgitation is a protective factor for left atrial thrombosis.

Objective To investigate the feasibility of ultrasound-guided percutaneous drainage for the treatment of peripancreatic abscess. Methods The clinical data of 36 patients with peripancreatic abscess who were admitted to the General Hospital of the Chengdu Military Command were retrospectively analyzed. All the puncture sites were designed according to the region, range and shape of the abscess, and then the angle and the direction of the needle penetration were determined according to the spatial relationship between the puncture site and the abscess. Finally, the drainage tubes were placed under the guidance of the ultrasound. Results The technique was successfully performed on all the patients, and 33 patients were cured with the cure rate of 92%.The mean healing time was 37 days. Three patients were converted to laparotomy because of the unsatisfied therapeutic effects. Enterocutaneous fistula was detected in 3 patients after the surgery and they were cured after receiving nonoperative management. All patients were followed up for 3-48 months, and neither residual abscess nor recurrence was detected. Two patients were complicated with type one diabetes, one with dyspepsia, two with gallstone, and they were cured by symptomatic treatment. The body weights of 27 patients were increased compared to those before the operation. Conclusion Ultrasound-guided percutaneous drainage is effective for the treatment of peripancreatic abscess.

Objective To investigate the effect of the peritoneal lymphatic stomata on intra-abdominal infection and the regulatory mechanism of angiotensin Ⅱ receptor specific inhibitor PD123319 on peritoneal lymphatic stomata.Methods The experimental study was adopted.Forty rats were divided into the control group,sham operation group,intra-abdominal infection group and intra-abdominal infection drug intervention group by the random number table,every group had 10 rats.The classic appendix perforation (CLP) intraabdominal infection model was established in the abdominal infection group.After establishing the model of abdominal infection,PD123319 solution was injected intraperitoneally immediately (0.2 g/kg) in the abdominal infection drug intervention group.Abdominal cavity of the rats in the sham operation group was opened,and then was shut after flipping the intestine.The rats in the control group,sham operation group and intra-abdominal infection group were treated with intraperitoneal injection of 1ml stroke-physiological saline solution.After 2 hours,the rats were sacrificed,and peritoneal tissue was taken for the following tests.(1) The aperture size and distribution density of peritoneal lymphatic stomata were observed by scanning electron microscope (SEM).(2) The nitric oxide (NO) concentration in the peritoneal tissues was detected using nitric oxide nitric acid reduction method.(3) The expressions of endothelial nitric oxide synthase (eNOS) and Phospho-eNOS (P-eNOS) were detected by the Western blot.(4) The intracellular Ca2+ concentration were detect by flow cytometry.Measurement data with normal distribution were presented as-x ± s.The comparison among groups was analyzed using the ANOVA and pairwise comparison was analyzed by the LSD test.Results (1) The aperture size and distribution density of the peritoneal lymphatic stomata in the control group,sham operation group,intra-abdominal infection group and intra-abdominal infection drug intervention group were respectively (2.3 ± 0.4) μm,(2.5 ± 0.5)μm,(4.7 ±0.5)pm,(3.8 ±0.5)pm and (2.0 ±0.8) × 108/m2,(2.1 ±0.7) × 108/m2,(6.2 ± 1.3) × 108/m2,(4.6 ± 1.4) × 108/m2,with statistically significant differences among the 4 groups (F =98.130,56.780,P ＜ 0.05).There were statistically significant differences in the aperture size and distribution density of the peritoneal lymphatic stomata between the intra-abdominal infection group and control group or intra-abdominal infection drug intervention group (t =11.586,8.573,3.854,3.098,P ＜ 0.05) and no statistically significant differences between the control group and sham operation group (t =1.281,0.514,P ＞0.05).(2) The concentrations of NO in the peritoneal tissues in the control group,sham operation group,intra-abdominal infection group and intra-abdominal infection drug intervention group were respectively (0.380 ± 0.024) μmol/gprot,(0.450 ±0.020) μmol/gprot,(1.253 ±0.033) μmol/gprot and (0.579 ±0.035) μmol/gprot,with a statistically significant difference among the 4 groups (F =52.725,P ＜ 0.05).There were statistically significant differences in the concentration of NO between the intra-abdominal infection group and control group or intra-abdominal infection drug intervention group (t =10.536,67.798,P ＜ 0.05) and no statistically significant difference in the concentration of NO between the control group and sham operation group (t =2.007,P ＞ 0.05).(3) The results of Western blot showed that the expressions of eNOS and P-eNOS in the control group,sham operation group,intra-abdominal infection group and intra-abdominal infection drug intervention group were respectively (0.591 ± 0.028)U/mg,(0.603 ± 0.007) U/mg,(0.615 ± 0.027) U/mg,(0.626 ±0.026) U/mg and (0.578 ±0.003)U/mg,(0.603 ± 0.071) U/mg,(0.773 ± 0.033) U/mg,(0.710 ± 0.012) U/mg,with no statistically significant difference in the expression of eNOS among the 4 groups (F =0.902,P ＞ 0.05) and with a statistically significant difference in the expression of P-eNOS among the 4 groups (F =205.062,P ＜ 0.05).There were statistically significant differences in the expression of P-eNOS between the control group and sham operation group or intra-abdominal infection group (t =7.678,13.322,P ＜ 0.05) and between the intra-abdominal infection group and intraabdominal infection drug intervention group (t =4.035,P ＜0.05).(4) The results of flow cytometry showed that Ca2+ concentration in the control group,sham operation group,intra-abdominal infection group and intraabdominal infection drug intervention group were respectively 82.200％ ± 0.060％,81.730％ ± 0.052％,21.980％ ± 0.010％,29.500％ ± 0.004％,showing a statistically significant difference between the 4 groups (F =21 271.030,P ＜ 0.05).There were statistically significant differences in the Ca2+ concentration between the intra-abdominal infection group and control group (t =164.750,P ＜ 0.05) and between the intra-abdominal infection group and intra-abdominal infection drug intervention group (t =21.338,P ＜ 0.05),and no statistically significant difference between the control group and sham operation group (t =1.861,P ＞ 0.05).Conclusion The intra-abdominal infection could increase aperture size and distribution density of peritoneal lymphatic stomata,and PD123319 may be through inhibiting the activation of NO synthase to decrease the concentration of NO,enhance the concentration of Ca2+ in peritoneal mesothelial cells and reduce the opening of peritoneal lymphatic stomata.

Objective To evaluate the immunoprotection by the inactivated whole bacteria(IWB) of Stenotrophomonas maltophilia K279a in mice.Methods Mice were immunized by inactivated whole bacteria of S.maltophilia K279a made from formaldehyde.When the indicated the antibody titer of the mice reached the require level, the protective effect of the IWB was evaluated by performing the opsonophagocytic killing test in vitro and the poison attack experiments in vivo. Results It was found that IgG in serum of the immunized mice measured by ELISA was significantly increased after the second immune enhancement, and antiserum in vitro had strong phagocytic effect.Meanwhile, immunoprotection of the immunized groups was also significantly increased when challenged by S.maltophilia K279a.Conclusion Effective humoral immune response can be predominantly induced by the inactivated whole bacteria of S.maltophilia K279a, providing protection against challenge by S.maltophilia K279a in BALB/c mice.

Objective To explore the interaction of streptococcus suis serotype 2 recombinant suilysin ( SLY ) with platelets, and provide the theoretical basis for clinic treatment of patients infected with S.suis.Methods The nickel column affinity chromatography was used to purify the recombinant SLY.The hemolytic acivity was identified by optical density before the platelets aggregation induced by a SLY was detected by a platelet aggregometer or electron microscope and the effect of aspirin on platelets aggregation was analyzed.The impact of wild type 05ZYH33 and sly-deficient mutant strainΔSLY on platelets of mice was compared to predict the interaction of the SLY with platelets in vivo.Results and Conclusion Hemolytic activity of recombinant SLY was 2000 hemolytic units( HU) and platelets aggregation was induced at 1 μg/ml.The aggregation can be inhibited by aspirin in 5 mmol/L.SLY can also increase the volume and reduce the amount of platelets in mice.