[Objective]To investigate the capability of synthesis BMP-2-derived peptide on osteogenic induction in bone marrow stem cells(BMSEs),and to evaluate the osteoinductivity of BMP-2-derived peptide in vitro.[Method]After segregating and cultivating four weeks Wistar rats marrow stromal cells in induction group were induced by osteogenasis medium containing 200 gg/ml BMP-2-derived peptide,and in non-inductional group BMSCs were still culture with DMEM medium.Following continue culture for 2~4 weeks,cells film preparation,alkaline phosphatase activity and calcium deposition were measured.Type I collagen(CoM)and osteopontin(OPN)mRNA expression were measured using real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)technique.The capability of synthesis BMP-2-derived peptide on osteogenic induction of BMSCs was investigated.[Result]After inductived with BMP-2-derived peptide.BMSCs cultured survived well greatly changed in cell morphology,and showed a biological and morphologie characteristics similar to those of osteoblasts.The lever of alkaline phosphatase activity and calcium deposition increased.Col-Ⅰand OPN mRNA were expressed at higher level.In non-inductional group no conspicuous osteogenic induction was shown.[Conclusion]BMP-2-derived peptide can induce BMSCs to differentiate into osteoblasts.It has the similar capacity of osteogenic induction as nature BMP-2,so BMP-2-derived peptide is an ideal cell agent for bone tissue engineering,and can be available widely.

Objective To study the effects of two fungal elicitors on the growth of Dendrobium candidum protocorms cultured on the solid medium.Methods The medium for propagation of protocorms was selected and the growth curve on that medium was obtained.According to the curve,the two fungal elicitors were added at the different growth stages of protocorms.The fresh weight(FW) and dry weight(DW) of protocorms were measured.Results The medium for propagation of protocorms was 1/2MS+20% potato extract+3% sucrose+0.8% agar.Compared with the control MF23 elicitor added at weeks 11 and 13 could significantly increase the FW by 16.4%(P

The efficacy of different kinds of insulin analogue administered by multiple subcutaneous injection or continuous insulin infusion was compared. The two forms of administration all can well control blood glucose. Continuous subcutaneous injection has better compliance.

Objective To observe the efficacy and side effects of Jianpi Qutan Decoction combined with Quetiapine in the treatment negative symptoms of schizophrenia.Methods 60 inpatients with schizophrenia were randomly divided into two groups,with one group treated with Jianpi Qutan Decoction combined Quetiapine and the other group treated with Quetiapine.Effects and side effects were assessed with the Positive and Negative Syndrome Scale (PANSS) and Treatment Emergent Symptoms Scale (TESS) respectively 1,4 and 8 weeks before and after the treatment.Results There was no significant difference in the overall efficacy between the two groups,but the improvement of the negative symptoms and illness provocation was significantly better in the treatment group than that in the control group respectively (P＜0.05).Moreover,the adverse reaction was milder in the treatment group.Conclusion It is suggested Jianpi Qutan Decoction in combination with typical antipsychotics is effective and safe in treating negative symptoms in schizophrenia patients.

Objective To observe the effect of metformin and exercise therapy for IGT. Methods 86 patients with IGT were given metformin and exercise therapy,the range of glucose,BMI and symptoms improvement were observed before and after the therapy. Results After 6 months therapy,the fasting blood glucose and 2 h after oral administration of 75 g glucose levels were significantly reduced(P＜0.01＝,BMI reduced(P＜0.05＝.72 cases (83.7%)were back to normal glucose tolerance,14 cases maintained IGT(those unable to adhere to exercise and no diet controlled),0 case became DM. Conclusion Metformin and exercise therapy had good efficacy in curing IGT.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced podocyte injury and its signal transduction mechanism.Methods Differentiated mouse podocytes were exposed to normal glucose,high glucose,and different concentrations of 1,25(OH)2D3 or LY294002 (a selective PI3K inhibitor) for 24 h.PCR and immunofluorescent staining were used to detect nephrin,podocin,and desmin.Western blotting was used to detect protein expression of nephrin,podocin,desmin,PI3K,Akt and p-Akt.Results Compared with high glucose group,1,25(OH)2D3 (100 nmol/L and 1000 nmol/L) significantly up-regulated the expression of podocin and nephrin in podocytes induced by high glucose (P ＜ 0.05).Meanwhile,1,25(OH)2D3 (100 nmol/L) significantly reduced the expression of desmin (P ＜ 0.05).PI3K and p-Akt were obviously reduced in high glucose group.In the presence of 1,25(OH)2D3,the trends were reversed.However the above effects of 1,25(OH)2D3 were abolished when p-Akt was blocked by the PI3K inhibitor LY294002.Conclusions 1,25 (OH)2D3 can inhibit high glucose-induced pedocyte injury through PI3K/p-Akt signaling pathway.

Objective To investigate the effect of active vitamin D (VD) on macrophage M1 and M2 phenotype and its role in protecting podocyte impairment in diabetic nephropathy (DN).Methods Diabetes mellitus rats were established by intraperitoneal injection with streptozocin.Rats were randomly divided into four groups:normal-1 (NC-1,n=8),normal-2 (NC-2,n=8,normal rats treated with calcitriol 0.1 μg· kg-1 · d-1 by gavages),DN (n=24) and VD (n=24,DN+calcitriol 0.1 μg· kg-1 · d-1 by gavages).Blood glucose and body weight were assessed,and 24-hour urine was collected regularly.Blood and urine samples were taken for biochemical study,and kidney tissues were used for PAS staining to assess histological changes.Immunohistochemical staining was used to detect number of CD68 + macrophage.Western blotting was used to detect protein expressions of nephrin,podocin,CD68,M1 specific marker of inducible nitric oxide synthase (iNOS),TNF-α and M2 specific marker of CD163,arginase 1 (Arg-1),mannose receptor (MR).Results (1) In DN group,levels of BUN,Scr,urinary protein and glomerular mesangial matrix proliferation were significantly higher (P ＜ 0.05),and the expressions of nephrin,podocin were significantly decreased compared with NC groups (P ＜ 0.05).These above changes were significantly improved in VD group (P ＜ 0.05).(2)Number of CD68 + macrophage infiltration in DN group was increased in a time dependent manner compared with NC groups,which was significantly reduced in VD group (P ＜ 0.05).(3)To further definite M1 and M2 macrophage activation phenotype,the protein expressions of iNOS and TNF-α was increased in DN group at 8th,14th,18th weeks compared with NC groups (P ＜ 0.05),which were significantly decreased in VD group (P ＜ 0.05).Although,there were no significant difference of protein expressions of CD163,Arg-1 and MR between VD and DN group at both 8th and 14th week (P ＞ 0.05),the protein expressions of CD163,Arg-1 and MR were higher in VD group at 18th week than that in DN group (P ＜ 0.05),and the ratio of CD163/CD68 was also enhanced in VD group (P ＜0.05).(4)Moreover,the protein expression of iNOS was negatively correlated with expression of either nephrin or podocin (r =-0.707,P ＜ 0.01; r =-0.712,P ＜ 0.01),whereas the protein expression of CD163 was positively correlated with expression of either nephrin or podocin (r =0.627,P＜ 0.01; r=0.613,P ＜ 0.01).Conclusion Vitamin D can regulate macrophage phenotype,via inhibiting M 1 macrophage activation and enhancing M2 macrophage activation to protect podocyte impairment.

Objective To investigate the effects and underlying mechanism of calcitriol on ameliorating podocytes impairment in DN rats.Methods SD rats were randomly divided into four groups:normal control (NC) group,calcitriol treatment (VD) group:calcitriol 0.1μg· kg--1 d-1,diabetic nephropathy (DN) group:streptozocin (STZ) 58 mg/kg,DN treated with calcitriol (DN + VD) group:calcitriol 0.1 μg · kg-1 · d-1 + STZ 58 mg/kg.Rats were sacrificed at the end of 18 weeks.Results Compared with the DN group,the DN + VD group exhibited significantly lower proteinuria by 36％,improved renal histology at the end of the experiment (P ＜ 0.05),and similar levels of blood glucose,serum urea nitrogen as well as body weight (P ＞ 0.05).There were no significant differences in the serum concentrations of creatinine,calcium and phosphorus among the four groups (P ＞ 0.05).In DN group,the expressions of nephrin,podocin,VDR,PI3K-p85 and p-Akt were significantly decreased and the expression of desmin was increased compared to NC group.Calcitriol treatment could attenuate the above changes.Additionally,a positive correlation was observed between the expressions of nephrin and VDR (r=0.776,P ＜ 0.05).Likewise,the expression of nephrin was positively correlated with either PI3K -p85 or p-Akt (r=-0.736,r=0.855,all P ＜ 0.05).Conclusion Calcitriol can ameliorate podocytes injury in DN rats,which might be related with the further up-regulation of PI3K/p-Akt signaling pathway.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced macrophage activation and its underlying signal transduction mechanism.Methods RAW 264.7 cells were used to perform cell culture,the activity of intracellular iNOS was measured.VDR siRNA and PPARγ antagonist pre-treatment with macrophages were done before using 10-8 mol/L1,25(OH)2D3 to intervene high glucose pre-incubated macrophages.M1 markers including iNOS,TNF-α,IL-12,M2 markers including MR,Arg-1,IL-10 and nuclear receptors VDR and PPARγ were separately examined.Results The iNOS activity was increased in a glucose-dose and time dependent manner.Particularly,25 mmol/L glucose at 24 h gave the maximum response.After being treated with 25 mmol/L glucose for 24 h,not only inflammatory cytokines of TNF-α,IL-12 in the supernatant were increased,but quantitative real-time PCR and Western blotting analysis showed iNOS was also up-regulated (P ＜ 0.05).However,M2 markers,i.e.MR and Arg-l were significantly decreased (P ＜ 0.05).When in the presence of 1,25(OH),D3,the trends were reversed:the markers of M1,including TNF-α,IL-12 and iNOS were obviously reduced (P ＜ 0.05),while M2 markers,IL-10,Arg-1 and MR were increased (P ＜ 0.05).In addition,VDR and PPARγ were also increased (P ＜ 0.05).However,the above effects of 1,25 (OH)2D3 were abolished when further inhibited the expression of VDR and PPARγby VDR siRNA and PPARγ antagonist.Besides,accompanied by VDR,PPARγwas also decreased upon the treatment with VDR siRNA (P ＜ 0.05).Conclusion 1,25(OH)2D3 can promote high glucose induced classically activated macrophages (M1) converting to alternatively activated macrophages (M2) and this is achieved through VDR-PPARγ pathway.

Objective To investigate the effect of moderate static magnetic fields (SMF) on secretion of inflammato?ry factors tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8) in human monocytic leukemic cell line THP-1. Methods THP-1 cells at logarithmic phase were divided into control group and magnetic treatment group. CCK-8 method was used to detect cell proliferation after THP-1 cells were exposed to 60 mT, 200 mT and 400 mT static magnetic fields at 18, 24 and 48 h. Then THP-1 cells were divided into control group, magnetic treatment group, LPS activation group and LPS+SMF treatment group. When magnetic treatment group and LPS+SMF treatment group were ex?posed to SMF at 18, 24 and 48 h, the levels of the cytokines TNF-α, IL-6 and IL-8 were determined by ELISA. Results (1) 60 mT, 200 mT and 400 mT SMF had no significant effects on cell proliferation in THP-1 cells (P>0.05). (2)THP-1 cells secreted more TNF-αand IL-6 in 24 h than 18 h in every group, while IL-8 didn′t change. Compared with 24 h, the secre?tion of TNF-αdecreased and IL-6 didn′t change, while IL-8 increased in 48 h. At three sampled time THP-1 cells of LPS activation group secreted more TNF-α, IL-6, IL-8 than those of control group and magnetic treatment group. After magnetic treatment THP-1 cells of LPS+SMF treatment group secreted less TNF-α, IL-6, IL-8 than those of LPS activation group (P<0.05). Conclusion Static magnetic field may have some inhibitory effects on release of TNF-α, IL-6, IL-8 from THP-1 cells, which can provide basic data for the treatment of rheumatoid arthritis.