Objective To explore curative efficacy of Xianlinggubao capsules combined with Lugua polypeptide injection in treatment of vertebral osteoporosis.Methods 90 patients of vertebral osteoporosis from March 2014 to March 2015 in our haspital were selected as research objects and randomly divided into the observation group and control group,with 45 cases in each group.The control group were treated with Lugua polypeptide injection, while the observation group were treated with Xianlinggubao capsules combined with Lugua polypeptide injection.Then bone mineral density, osteocalcin, visual analogue scale (VAS), Oswestry disability index (ODI) score, curative effect in two groups after treatment were compared.Results Ater treatment, bone mineral density, osteocalcin in observation group was than that in control group [(0.99 ±0.13)g/cm2 vs(0.89 ±0.14)g/cm2, (41.05 ±5.30)μg/L vs(35.80 ±5.23)μg/L],the difference was statistically significant(P<0.05); VAS、ODI score in observation group were lower than those in control group[(1.21 ±0.30)分vs(4.65 ±1.08)分、(18.24 ±4.97)% vs(31.26 ±5.73)%],the difference was statistically significant (P<0.05).The total effective rate of observation group was statistically higher than that in the control group 95.55%(43/45),the difference was statistically significant vs77.77%( 35/45 ) ( P<0.05 ) .Conclusion Xianlinggubao capsules combined with Lugua polypeptide injection is well for vertebral osteoporosis,which can improve the bone metabolism of patients, promote bone growth and improve treatment efficacy.

Objective To assess the feasibility and effectiveness of percutaneous transluminal angioplasty (PTA)for the salvage of immature arteriovenous fistula (AVF) and to identify the incidence of arterial and venous puncture site spasm.Methods The medical records and radiological data of 88 patients with 112 interventional procedures for immature AVFs were retrospectively reviewed.Results The stenosis lesions were (2.0 ± 1.4) cm long.Technical success rate and clinical success rate were 80.4％ (78/97) and 92.8％ (90/97) for PTA via brachial artery,85.7％ (6/7) and 100％ (7/7) for PTA via vein,25％ (2/8) and 50％ (4/8) for PTA via both brachial artery and vein,respectively.Spasm of pure arterial PTA occurred in 2 patients (2.1％) and was mild and moderate.Spasm of pure venous PTA occurred in 2 patients (28.6％) and was both moderate.Spasm of combined arterial and venous PTA occurred in 3 patients (37.5％) and from being severe to completely occluded.By comparison,there were statistical differences of technical and clinical success rate (P =0.000,0.019 ; P =0.000,0.029),fistulas spasm rate was statistically significant different (P =0.000).Conclusions Endovascular therapy was effective in restoring the dysfunctional native AVFs,it was safer and more effective and with less sideeffects especially in selecting coronary balloon to treat patients without large phlebangioma and round fistulas.

After the management models of chronic diseases were described and the disadvantages of traditional management model of chronic diseases and Internet + management model of chronic diseases were summarized, the whole course management-basedInternet+ management model of chronic diseases was proposed according to the studies on and the implementation of Internet + management model of chronic diseases with its connotations studied in order to better service its users.

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.

Objective To study the molecular characteristic of the epidemic plasmids carrying blaCTX-M-15 in Guangzhou. Methods A total of 38 strains of E. coli and 47 strains of K. pneumoniae both producing CTX-M-15 ESBLs were collected from nine hospitals in Guangzhou from 2007 to 2008. The clonal relationship of isolates carrying blaCTX-M-15 was determined by PFGE and MLST. Antimicrobial susceptibility was determined by microdilution test for all isolates. Conjugative plasmids carrying blaCTX-M-15 were obtained by mating and were subject to restriction analysis. PCR was used to determine phylogenetic groups of E. coli,and to study replicon type and the genetic contexts of the plasmids harboring blaCTX-M-15. Serum agglutination test was used to detect the serotype of E. coli. Results The 37 strains of E. coli were classified into 28 genotypes, while the 47 strains of K. pneumoniae were divided into 30 genotypes. ST131 was found in E. coli but not O25 serotype. Two novel-alleles of tonB and new ST were determined in K. pneumoniae. Forty out of 58 isolates represented independent genotypes have been succeeded to transfer the plasmid carrying blaCTX-M-15 to the E. coli C600(Rif) by conjugation. The sizes of plasmids carrying blaCTX-M-15 are 65 kb in 57.9% isolates of E. coli and 92 kb in 87.5% isolates of K. pneumoniae. Two epidemic plasmids were detected in E.coli and K. pneumoniae by restriction enzyme, designated p15-e and p15-k respectively. The blaCTX-M-15 and ISEcpI were identified on p15-e, and the blaCTX-M-15 ,ISEcpI,aac(6')- Ⅰ b,aac(3')-Ⅲ ,blaOXA-1 ,qnrB,qnrS,blaDHA-1 , blaTEM-1 were determined on p15-k. The p15-k also was identified to belong to the incompatible group FⅡ. Conclusion The local dissemination of blaCTX-M-15 appears to be due to the spread of epidemic plasmids harboring blaCTX-M-15. No evidence supports the dissemination of clone strains which carried blaCTX-M-15.

Objective To investigate the impact on the resistance of carbapenem with the expression of OprD2 or OprD2 mutation in Pseudomonas aeruginosa. Methods One hundred and one clinical strains of Pseudomonas aeruginosa with MIC for imipenem ≥8 μg/ml were studied. MIC were determined by the broth microdilution method, and the antibiotics tested were imipenem(IPM ), biapenem( BPM), meropenem(MEM) and panipenem(PEM). The expression of the oprD2 gene in Pseudomonas aeruginosa were analyzed by real-time reverse transcriptase PCR(RT-PCR). For the Pseudomonas aeruginosa with normal expression of OprD2 and resistance to imipenem, full-length oprD2 gene was amplified by PCR and the products were sequenced. Results According to the result of the expression of oprD2 gene, 101 strains of Pseudomonas aeruginosa were divided into two groups: group1 with diminished expression of OprD2, and group2 with normal expression of OprD2. Comparing isolates with MIC of 4 kinds of carbepenem agents ≥ 16 μg/ml in two groups. Data showed the amount of OprD2 expression were different between two groups(P ＜0.01 or P ＜ 0.05). In group1, there are 28 isolates with MIC ≥ 16 μg/ml of all the 4 kinds of carbapenems, among which 25 isolates have obviously diminished expression of OprD2 ( ＜ 0.4). Negative correlations tendency appeared between the level of OprD2 transcription and MICs of 4 kinds of carbepenem agents in Pseudomonas aeruginosa. In group2, 16 strains with OprD2 mutation divided into 4 types according to the pattern of alteration. Compared with PAO1, these strains have increased MIC with different degree to IPM,BPM, MEM and PEM. Conclusion The deletion or diminished expression of OprD2 resulted in resistance to imipenem in Pseudomonas aeruginosa. The level of OprD2 transcription and antimicrobial activities for carbapenem agents proved to be highly correlated in Pseudomonas aeruginosa. The mutation of OprD2 in Pseudomonas aeruginosa probably decreased the sensitivity of carbapenem agents against Pseudomonas aeruginosa.

BACKGROUND:A preliminary experiment developed a double-chamber stirred bioreactor which can carry out osteogenic and cartilage induction at the same time.
OBJECTIVE:To explore the effects of double-chamber stirred bioreactor on the repair of goat knee cartilage defects with tissue-engineered cartilage.
METHODS:Twelve goats were selected to make bilateral femoral condyle osteochondral defects models and randomized to three groups:experimental group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor under mechanical stimulation;control group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor;blank control group, without treatment. After 12 and 24 weeks of implantation, general observation, Masson staining, II col agen immunohistochemical staining and histological scoring were performed.
RESULTS AND CONCLUSION:In the experimental and control groups, new cartilage tissue and bone tissue were visible, but the experimental group showed better repair effects than the control group (P<0.05). The blank control group had no cartilage formation. These findings indicate that under the mechanical stimulation by the double-chamber stirred bioreactor in vitro, the repair effect of tissue-engineered osteochondral complex on knee joint cartilage defects can be improved.

Objective To study the role of deleted in liver cancer-1 (DLC-1) gene main domains on the regulation of human colon cancer HT29 cell proliferation.Methods Subcloning recombinant plasmid vectors with Rho GTPase activating protein (RhoGAP),sterile alpha motif (SAM) or steroidogenic acute regulatory-related lipid-transfer (START) domains of DLC-1 gene knockout were constructed and transfected into human colon cancer cell HT29.Wild HT29 cell group (control group),pcDNA3.1-HT29 cell group (vector group) and pcDNA3.1-HT29-DLC-1 cell group (whole DLC-1 gene transfected group) were set as control.The change of cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and colony formation test.The cell apoptosis was analyzed by flow cytometry.The activity of RhoA protein was detected by pull-down assay.The differences between the groups were analyzed by the analysis of variance.Results At 48 hours after the successful transfection,compared with control group and vector group,cells proliferation and the activity of RhoA protein were significantly suppressed in whole DLC-1 gene transfected group (F=146.36,698.08,both P＜0.05) and early cell apoptosis increased (F=294.08,P＜0.05).Compared with control group and vector group,there was no significant difference in cell proliferation ability,cell apoptosis and the activity of RhoA protein activity in RhoGAP knockout transfected cells (F=0.99,0.049,5.769,all P＞0.05).Compared with whole DLC-1 gene transfected group,the suppression of cell proliferation was more significant in SAM knockout transfected cells (F=31.00,P＜0.05),the activity of RhoA protein down regulated (F=92.57,P＜0.05) and apoptosis increased (F=130.44,P＜0.05).Compared with whole DLC-1 gene transfected group,the ability of cell proliferation increased (F=15.47,P＜0.05),apoptosis cell decreased (F＝110.23,P＜0.05) and the activity of RhoA protein up regulated (F=199.39,P＜0.05) in START knockout transfected cells.Conclusions The role of DLC-1 gene in the suppression of cell proliferation in HT29 cells was RhoGAP-dependent.SAM domain may be the self suppression domain for endogenous RhoGAP activity.START domain may take effect through enhancing RhoGAP domain.

Objective To explore the prognostic value of Acute Kidney Injury Network (AKIN)criteria combined with Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ)and Sequential Organ Failure Assessment(SOFA)scoring system in acute kidney injury(AKI)after cardiac surgery. Methods Clinical data of patients who underwent open-heart surgery in Zhongshan Hospital,Fudan University from April 2009 to August 2009 were prospectively collected.AKI after cardiac surgery was classified by AKIN staging system.APACHE Ⅱ and SOFA scores were evaluated according to the worst value of physiologic variables in the 1st 24 h after surgery.Discrimination and calibration of these three models were assessed by receiver operating characteristic(ROC) curve and Hosmer-Lemeshow goodness-of-fit test.Besides,their effects on inhospital mortality were evaluated by multivariate Logistic regression analysis. Results Of the 993 admissions,309 patients developed AKI and the incidence was 31.1%.The median time that developed postoperative AKI and reached the Scr peak were 1 d and 2 d respectively.Either APACHE Ⅱ or SOFA scores,which was positively correlted with the severity of AKI(APACHEⅡr=0.37,P<0.01;SOFA r=0.42,P<0.01)was higher in AKI patients compared with that in nonAKI patients(P<0.01).The mortality rose corresponding to the severity of kidney injury.However,the predicted death rate-adjusted(PDR-A)calculated by APACHE Ⅱ scoreS Was higher than the actual value in non-AKI patients and AKIN stage 1(P<0.01),while it was lower in AKIN stage 3 (P<0.01).The areas under the ROC curve of APACHEⅡ,SOFA and AKIN criteria were all above 0.8 and the results of Hosmer-Lemeshow goodness-of-fit test indicated good calibration of three models.Multivariate analysis showed that APACHE Ⅱ≥19(OR=4.26)and AKIN stage 3(OR=76.151 were independent predictors of in-hospital mortality. Conclusions AKI can be classified by AKIN criteria in the early stage after cardiac surgery and the AKIN staging system may serve the prediction of prognosis.The APACHE Ⅱ and SOFA scores just evaluated in the 1st 24 h after operation can discern the severity of patients'illness.Three models all present good discrimination and calibration in predicting patients'outcome.APACHE Ⅱ≥19 along with AKIN stage 3 are found to be the independent predictors of in-hospital mortality.It should be noticed that the deviation between PDR-A and the actual mortality in subgroups,dynamic evaluation may raise the accuracy of scoring system.