Objective To explore the correlation between the change of intestinal flora and insulin resistance in pre-diabetes population,and the role of intestinal flora in the occurrence and development in the pre-diabetes population.Methods two hundreds and fifty cases of pre-diabetes in our hospital were selected and divided into the group A(pre-diabetes intervention group)and B(prediabetes non-treatment group).Fifty cases of diabetes(positive control group)served as the group C.The cohort study was adopted.The follow-up intervention lasted for two years.The intestinal flora and blood biochemical indicators were detected in different groups and at different time periods.The insulin resistance index was calculated.Results The total bacterial count before intervention in the group A and B was significantly higher than that in the group C;Enterococcus genus in the group C was significantly higher than that in the group A and B;the proteus and lactobacillus content in the group A and B was higher than that in the group C,but the difference was not significant;Bifidobacterium genus in the group C was higher than that in the group A and B.After intervention,the total bacterial amount in the group Awas significantly higher than the group B and C,but there was no statistical difference between the group B and C;enterococcus genus in the group C was significantly higher than that in the group A and B,enterococcus genus after intervention in the group A was reduced and which in the group B was increased,the difference had statistical significance;the proteus and lactobacillus content in the group A and B was higher than that in the group C,but the difference was not significant;bifidobacterium genus in the group C was higher than that in the group A and B,moreover which after intervention in the group B was increased,the difference was statistically significant.The insulin resistance before processing had no statistical difference between the groups A and B,which in the group C was higher than the group A and B,the insulin resistance after intervention in the group A was decreased,the difference was statistically significant.The total bacterial count,enterococcus and bifidobacterium were positively correlated with insulin resistance,and bacteroides,proteus and lactobacillus had no significant correlation with insulin resistance.Conclusion Intestinal flora has correlation with insulin resistance,and serves as a marker of pre-diabetes development,implementing the probiotics adjustment combined the quantization aerobic muscular movement can realize the adjustment of insulin sensitivity and improves the diabetic development in the prediabetic population.

Objective To study the possible relationships between expression of matrix metalloproteinase(MMP)2,9 and the pathogenesis of preeclampsia in which trophoblast invasion is impaired. Methods MMP-2,9 expression were detected by immunohistochemistry streptavidin-biotin complex (SABC)method in 20 normal term placentae and 20 preeclampsia placentae,respectively.In addition, mRNAs for MMP-2,9 were analyzed by real time PCR in both groups.Results The intensities of both MMP-2 and MMP-9 immunostaining in preeclampsia placentae were significantly declined compared to those of normal term placentae(P

Objective To test the hypothesis that homocysteine can decrease MMP-2 and MMP-9 expression in cultured trophoblasts of early pregnancy and that homocysteine can prevent trophoblasts invasion in the early stage of preeclampsia.Methods Cytotrophoblasts from early pregnancy were isolated and cultured.Trophoblasts were treated with or without Hcy(1 mmol/L)for 48 hour,and real time RT-PCR and gelatin zymography were used to quantify the mRNA and protease activity of MMP-2,-9.Results Treatment with Hcy(1 mmol/L)induced a decrease in MMP-2 mRNA by 21% and MMP-9 mRNA by 11%.At protein level MMP-2 expression decreased 14% and MMP-9 expression decreased 52% compared with control.Conclusions Homocysteine can decrease MMP-2,-9 expression in trophoblasts of early pregnancy and influence its invasion process.

Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20:sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20: sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20:sTRAIL were secreted from MSC. scFvCD20:sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20:sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20:sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20:sTRAIL, pLenR. ISZ:sTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20:sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20:sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20:sTRAIL,and thus locally concentrated scFvCD20:sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20:sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .

Objective To observe the effect of early rehabilitation therapy on recovery from reoperation for recurrent lumbar disc herniation (RLDH).Methods Sixty-five cases who received surgery for RLDH between 2007 and 2009 were randomly divided into a rehabilitation group and control group.Both groups were treated with the same surgical approach and routine treatment.Early and comprehensive rehabilitation therapy was provided in the rehabilitation group during the perioperative period,including preoperative and postoperative muscle strength training,postoperative sitting and standing balance training,and acupuncture.The control group was instructed only in general exercise.Before the operation and 2 weeks and 3,6,12 and 24 months afterward,the surgical outcomes of all cases were assessed using the JOA score and the improvement rate in the JOA score.Any postoperative complications and intervertebral fusion were also observed.Results The average postoperative JOA scores of both groups were significantly higher than their preoperative scores.At all of the time points after the operation,the average JOA scores and all improvement rates in the rehabilitation group were significantly higher than those in the control group.Postoperative complications such as deep venous thrombosis,urinary retention and constipation were significantly less among the rehabilitation group than among the controls.All the intervertebral bone implants were well fused on time.Conclusion Early rehabilitation can significantly improve the effectiveness of RLDH reoperation and reduce the incidence of postoperative complications.It is recommended for clinical application.

Objective To evaluate the neuroprotective effect and safety of neonatal hypoxic-ischemic encephalopathy(HIE)treated with recombinant human erythropoietin(rhEPO).Methods Fifty-three neonates with HIE were randomly divided into rhEPO treated group(n=29) with the dosage of 300 U/(kg?time),three times a week for 2 weeks and control group(n=24)without rhEPO.All supportive measures were same between 2 groups.Neurological scoring was evaluated at d3,d5 and d7 Neonatal behavioral neurological assessment(NBNA) was evaluated at d7,d14 and d28.The neurodevelopment quote was evaluated at age of 3 and 6 months.Blood pressure,liver and renal function,blood electrolytes and blood hemoglobin,platelet and reticular red blood cell count were monitored before and after treatment in all infants.Results The neurological scoring between two groups had no difference at d3.The significant difference was found at d7(P0.05).Conclusions Teraphy with rhEPO on neonatal HIE infants can promote neurological recovery,and there is no serious side effect with rhEPO treatment.

Aim To look for novel small-molecule inhibitors of CDK9 through structure-based virtual screening and biological activity determination.Methods Homology modeling of CDK9 was based on the 3-D structure of other cyclin-dependent kinase family members,and then virtual screening by DOCK(molecular docking)of database of small molecule was carried on.MTT method was used in inhibition of tumor cell growth in vitro,while Western blot was used for further study of molecular mechanisms.Results From the top 1000 compounds with the best DOCK energy score,27 compounds were selected for biological assay based on the diversity of chemical structure and functional group.12 of 27 selected compounds showed significantly inhibition activity on tumor cell proliferation,and only one compound in 12 with half-maximum inhibition concentration(IC50)values less than 20 ?mol?L-1 named C-21 was selected for further molecular mechanism study.The western blotting data showed C-21 compound could effectively inhibit CDK9 from phosphorylating large subunit C-terminal of RNA polymerase Ⅱ in a dose-dependent manner.Conclusions Through homology modeling,virtual screening by computer,determination of biological activity and experimental studies of molecular mechanism,a new promising lead compound targeted for CDK9 was found and confirmed.

Xylanase and mannanase are two hemicellulases,which are widely used in many fields.To improve the expression of thermostable xylanase DSB and thermostable mannanase ManA in Pichia pastoris,three endogenous signal peptides of Pichia pastoris(Scw1 1,Dse4 and Exg1) were chosed.Their capability to mediate the secretion of DSB and ManA with that of the Saccharomyces cerevisiae α-factor were compared.In shake-flask cultivation,three endogenous signal peptides and α-factor efficiently mediated the secretion of DSB and ManA,but the secretion efficiency has obvious difference.As for DSB,the expression efficiency of α-factor was much higher than three endogenous signal peptides.But as for ManA,the expression efficiency of Dse4 was equal to α-factor and much higher than Scw11 and Exg1.Therefore,α-factoris the most efficient signal peptide for DSB expression and Dse4 or α-factor are the most efficient signal peptide for ManA expression in Pichia pastoris X33.Moreover,the intracellular activities of DSB and ManA by α-factor are higher than Scw1 1,Dse4 and Exg1,and the intracellular activity of ManA was higher than DSB (the molecular weight of ManA was larger than DSB).Thus,when ManA were expressed in Pichia pastoris,different signal peptide,such as Dse4,could be used for improving the secretion efficiency.A basis for identifying more available signal peptides and screening for the optimal signal peptide for the target protein was provided.

Objective To evaluate the effects of asymptomatic arteriovenous fistula closure on left ventricular morphology and function in renal transplant recipients.Methods Between March 2007 and March 2011,a total of 60 patients undergoing consecutive kidney transplantation with asymptomatic arteriovenous fistula were divided randomly into two groups: arteriovenous fistula closure group,and non-arteriovenous fistula closure group.By using echocardiography,the changes in CO,CI,EF,LVEDV and LVMI were analyzed.Results At 12th month after transplantation,the values of CO,LVEDV and LVMI were significantly lower than those before transplantation (P＜0.05).The value of CI also showed a tendency to decrease (P＞0.05),and the value of EF was increased significantly (P＜0.05).At 6th month after arteriovenous fistula closure (18 months after transplantation),the values of CO,CI,LVEDV and LVMI were significantly lower than those before arteriovenous fistula closure (12 months after transplantation) (P＜0.05),and the value of EF was increased significantly (P＜0.05),but the values of CO,CI,EF,LVEDV and LVMI remained unc(b)anged in controls (P＞0.05).At 18th month after transplantation,the values of CO (4.4 ±0.8 L/min),CI [3.0 ± 0.8 L·min-1·m-2],LVEDV (110.0 ± 17.4 ml) and LVMI (114.7 ± 42.5g/m2) in trial group were significantly lower than the values [CO: 5.1 ± 0.9 L/min,CI: 3.5 ± 1.0L·min-1·m-2,LVEDV: 121.4±19.3 mL,LVMI: 138.4±44.1 g/m2] in controls (P＜0.05),and the value of EF (75.2％ ± 7.4％ vs.70.5％ ± 8.2％) significantly higher (P＜005).Conclusion In both groups,kidney transplantation benefits significantly the regression of cardiac mass,cardiac index and left ventricular dimensions,but closure of asymptomatic AVF induces more significant regression.

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.