Objective To explore the detailed underlying molecular and signaling mechanisms in the effects of icariin on bone formation by an in vitro cell model. Methods The proliferation of MC3T3-E1 osteoblast-like cells was evaluated by MTT, and gene expression of cell cycle related proteins in MC3T3-E1 cells after icariin treatment was detected by real-time PCR. The phosphorylation of MAPK signals, including ERK, P38, and JNK was determined by Western blot, and then the inhibitors of MAPK signals were used to treat cells with icariin alone or together to determine the role of MAPKs in the process of icariin treatment on MC3T3-E1 cell proliferation. Alkaline phosphatase and Alizarin red staining were used to detect the formation of mineralization nodules, and gene expressions of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in osteoblasts after being treated by icariin were evaluated by real-time PCR. ICI182780, and nilutamide was used to decide the participation of estrogen and androgen receptor signals in the process of icariin treatment on the differentiation and mineralization of MC3T3-E1 cells. Results Treatment with icariin promoted MC3T3-E1 cell growth in a time- and dose-dependent manner. This treatment also revealed that icariin increased the expression of mRNAs encoding both cyclin E and PCNA, positive regulators of cell growth, but decreased levels of mRNAs encoding Cdkn2b, a negative regulator of cell cycle progression. When MC3T3-E1 cells were cultured in a differentiated condition, icariin enhanced mineralized nodule formation and increased the expression of mRNAs encoding alkaline phosphatase, type Ⅰ collagen, and osteocalcin. Treatment with icariin significantly induced phosphorylation of both ERK and JNK and this phosphorylated effect occurred very rapidly within 5 minutes and reached peak at 15 minutes. Furthermore, the stimulated effects of icariin on proliferation and gene expression of cyclin E, PCNA, and Cdkn2b in MC3T3-E1 cells were dramatically attenuated by treatment with both U0126 and SP600125, inhibitors of MAPKs. Interestingly, such stimulating effects of icariin were at least partly reduced by treatment with ICI182780, an inhibitor of estrogen receptor. Icariin induced mineralized nodule formation and gene expression of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in MC3T3-E1 cells were also partly reduced when the cells were treated with ICI182780. Conclusions Our findings indicate that the anabolic effect of icariin on bone formation is, at least partly, mediated through the MAPK signaling pathway in order to modulate osteoblast proliferation and differentiation.

Objective The aim of this study was to explore the effect of low density lipoprotein ( LDL) on the proliferation and differentiation of MC3T3-E1 osteoblasts, as well as the expression of low density lipoprotein receptor-related protein 5(LRP5) and dickkopf-1(DKK1) mRNA of MC3T3-E1 osteoblasts. The possible mechanisms of the treatment of atorvastatin on LDL expression in MC3T3-E1 osteoblasts were also investigated. Methods Proliferation, osteocalcin expression, LRP5, and expression of DKK1 mRNA of MC3T3-E1 with interaction of LDL at 0. 05, 0. 10, 0. 20 mg/ml levels after 24 h, 48 h, 72 h were detected by CCK8, ELISA, and fluorescence quantitative PCR. Furthermore, proliferation, osteocalcin expression, LRP5 and DKK1 mRNA of MC3T3-E1 after the treatment of atorvastatin of 10-6 mol/L and 10-5 mol/L were also be studied, respectively. Results The effect of LDL on proliferation, expression of osteocalcin and expression of LRP5 and DKK1 mRNA in MC3T3-E1 osteoblasts was the most obvious under LDL with 0. 20 mg/ml level. Under that level, atorvastatin (10-6 mol/L or 10-5 mol/L) was able to make the proliferation of MC3T3-E1 osteoblasts in 48 h and 72 h be decreased, while significantly caused upregulation of osteocalcin, LRP5 mRNA expression; and down regulated DKK1 mRNA expression ( all P<0. 05). Conclusions Atorvastatin can reduce the inhibitory effect of LDL on the proliferation and differentiation of osteoblasts. The mechanisms of atorvastatin on osteoblasts are possibly related to the osteoblast proliferation and expression of LRP5 mRNA and DKK1 mRNA of osteoblasts of wnt signal pathway.

Baicalein (BAI) is an effective bactericide. The antibacterial activity and mechanism experiments were carried out by determining conductivity and content of macromolecules of membrane penetrability, the oxidative respiratory metabolism and protein synthesis changes and the inhibition of DNA topoisomerase activities. Electrical conductivity and the number of large molecules of BAI increased 2.48% and 1.8%, respectively, than that of the control. However, the membrane integrity did not destroyed by BAI directly. With BAI treatment, inhibition rates of activities for SDH and MDH were 56.2% and 57.4%, respectively, demonstrating that BAI could inhibit cell respiratory. After treated with BAI for 20 h, the total soluble content of proteins decreased by 42.83%. Moreover, the activities of DNA topoisomerase I and II were inhibited completely by 0.2 mmol x L(-1) BAI. These results indicated that BAI had obvious antibacterial activity on Staphylococcus aureus. The mechanism is that it could affect bacterial membrane penetrability, inhibit protein synthesis and influence SDH, MDH and DNA topoisomerase I and II activities to exert its antibacterial functions.

Objective To compare the efficacy of reversed less invasive stabilization system (LISS) and intramedullary fixation devices for treatment of femoral trochanteric fractures.Methods Data of 362 consecutive patients with femoral trochanteric fractures who were treated with reversed LISS or intramedullary fixation devices at our institution between March 2004 and May 2011 were retrospectively analyzed.There were 32 males and 38 females treated with reversed LISS.The mean age at injury was 73.4 years.There were 125 males and 167 females treated with intramedullary fixation.The mean age at injury was 74.7 years.The operation time,intraoperation blood loss and length of hospitalization were compared.The patients were asked to fill in a questionnaire of Harris hip score,and radiographs were used to evaluate the bone healing situation.Results The mean operation time was 120 min in reversed LISS group and 80 min in intramedullary group.The length of hospitalization was 12 days in reversed LISS group and 10 days in intramedullary group.More operation time and longer length of hospitalization were needed in reversed LISS group.The intraoperation blood loss was 100 ml in reversed LISS group and 100 ml in intramedullary group.There were 3 DVT in reversed LISS group,10 DVT and 3 PE in intramedullary group.No difference was found in the aspect of intraoperation blood loss,postoperative deep venous thrombosis and pulmonary embolism rate.A total of 320 patients were successfully followed-up,including 63 in LISS group and 257 in intramedullary group.The median Harris score was 75 in LISS group and 77 in intramedullary fixation group respectively.There was no difference of hip function score between two groups.The implant-related complication rate was 11.1％ in reversed LISS group,including 7 implant breakages.In intramedullary fixation group,there were neck screw exiting in 2 patients and cutout in 9 patients.So the implant-related complication rate was 4.3％ in intramedullary fixation group,which is statistically lower.Conclusion Both reversed LISS and intramedullary are effective for the treatment of proximal femoral fractures.There were no major difference in functional outcome between LISS and intramedullary nail.Intramedullary nail is still the choice of priority in most unstable proximal femoral fractures.The implants related complications in reversed LISS group are higher than Intramedullary nail group.However,for the unstable fractures proximal femoral fractures with lateral wall fracture,in which nailing may be difficult,reversely using LISS may be a good alternative.

Objective To explore the relationship between intermedin (IMD) and renal interstitial capillary loss in IgA nephropathy (IgAN) patients.Methods Renal biopsy specimens collected from primary IgAN patients in our hospital (n=80) were compared with normal renal tissues.Expressions of IMD,CD31 and VE-cadherin were examined by immunohistochemical method,and plasma concentrations of IMD and TGF-β1 in 37 cases from the 80 cases were compared.The relationship between IMD and renal interstitial capillary loss in IgAN patients was analyzed.Results IMD and VE-cadherin in renal tubule interstitium expressions increased compared to the control group at the early stage of IgAN (P ＜ 0.05).CD31 expression remained unchanged at the early stage of pathological lesions of IgAN (P ＞ 0.05),but decreased at the early stage of clinical stage of IgAN compared to the control (P ＜ 0.05).Expressions of IMD,CD31 and VE-cadherin were reducing as the disease progressed,and the correlations of CD31 and VE-cadherin (r=0.517,P ＜ 0.01),IMD and CD31(r=0.655,P ＜ 0.01) or IMD and VE-cadherin (r=0.576,P ＜ 0.01) were positive.Plasma concentrations of IMD and TGF-β1 were higher than those of the control group at the early stage of IgAN (P ＜ 0.05),and the changes of IMD and TGF-β1were correlated positively (r=0.582,P ＜ 0.01).Conclusion Compared with the control group,expression of IMD in kidney tubules increases at the early stage of IgAN,and change of IMD correlates closely with the renal interstitial capillary loss.Plasma concentrations of IMD and TGF-β1 increase compared with the control group at the early stage of IgAN,and the changes of IMD and TGF-β1 are related closely.

Objective To investigate the glomerular microvascular injury and repair in patients with IgA nephropathy (IgAN) as well as its relationship with intermedin (IMD).Methods Eighty cases of renal tissue taken from patients first diagnosed as IgAN in Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University and 15 cases of normal renal tissue were detected by the expression of glomerular IMD,CD31,and VE-cadherin through immunohistochemical method.ELISA method was used to detect VEGF and IMD of plasm from 31 normal subjects and 36 cases chosen from the IgAN patients.Their changes and internal relationship were analyzed according to Lee's and chronic kidney disease (CKD) classification.Results (1) Compared with the control group the expressions of CD31,IMD,and VE-cadherin in IgAN patients were statistically significant (P ＜0.01).Compared with the control group the levels of IMD and VEGF in plasma of IgAN patients in early stage of CKD group and late stage of CKD group were statistically significant (P ＜ 0.01).(2)Correlation analysis:the expression of glomerular CD31 and Lee's classification were negatively correlated (r=-0.232,P ＜ 0.05);glomerular IMD was negatively correlated with Lee's classification (r=-0.241,P＜0.05),while positively correlated with glomerular VE-cadherin (r=0.417,P＜ 0.01).VEGF in plasma of IgAN patients was positive correlated with CKD classification,BUN (r=0.458,0.409,P＜0.05),and negatively correlated with serum ALB (r=-0.532,P＜0.01).Conclusion Microvascular injury exists in patients with IgAN.The expression of VE-cadherin and IMD are positively correlated,suggesting that IMD may be involved in the progression of vascular protection and angiogenesis in IgAN.The contents of IMD and VEGF in plasma of IgAN patients increase,indicating that they may play a role in the progression of IgAN.

Objective To investigate the effects of icarrin on the activity and protein expression of core binding factor otl(Cbfa1) in rat osteoblasts cultured in vitro,and to explore whether mitogen-activated protein kinase (MAPK) pathway is involved in this process.Methods Calvarial osteoblasts were obtained from newborn (<24 h) SD rats by trypsin-coUagenase digestion method.The second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) or estradiol (10-8 mol/L) with or without extracellular-signal regulated kinase (ERK) inhibitor (UO126) or p38MAPK inhibitor (SB203580).Nuclear protein was extracted from osteoblasts.And then the activity of Cbfa1 was detected by ELISA.The amounts of Cbfa1 protein were detected by Western blot.Results Calvarial osteoblasts were obtained successfully and were used in this study after indentified by alkaline phosphatase and mineralized nodus staining.Cbfa1 expression and the activity in osteoblasts were up-regulated by both icariin and estradiol (P<0.05).The effects were partly inhibited by addition of U0126or SB203580 (P<0.05).Conclusions Either icarrin or estradiol can stimulate the proliferation and maturation of cultured osteoblasts in vitro via up-regulating the activity and expression of Cbfal.The MAPK signal pathway inhibitor seems to partly decrease Cbfa1 activity.It suggests that MAPK pathway may be involved in the transduction of icariin's impact on proliferation and mineralization of osteoblasts.

Objective To investigate the expression and clinical significance of tumor infiltrating dendritic cells(TIDCs) within gastric tumor tissues.Methods Immunohistochemistry(IHC),in-situ hybridization(ISH) and flow cytometry were applied to detect the expression of S100,CD83 mRNA and CD83 on DCs in 45 gastric adenocarcinoma tissues.The co-relationship of the S100 and CD83 expression with clinical(pathological) features was analyzed.Results IHC showed that S100 expression was unevenly distributed(within) 42 cancer tissue and CD83 was only expressed in tumor-adjacent tissue and normal tissue.ISH showed that CD83 mRNA was limitedly expressed within 7 samples.S100 expression had no significant(correlation) with clinical pathological features,while CD83 reversely correlated with TNM stages.Detected by flow cytometry,CD83 was expressed in low level in all 45 gastric cancer tissues and negative correlations were found with lymph node metastasis and TNM stages of gastric cancer(r=-0.879,P

BACKGROUND: Pedicle screw fixation is a common repair method for thoracolumbar fractures. Of them, pedicle screw placement into the injured vertebra is an important method. However, the long-term effects and the related biomechanical properties deserve further analysis. OBJECTIVE: To analyze the long-term effect and biomechanical characteristics of the internal fixation of thoracolumbar fractures by pedicle screw implantation. METHODS: A retrospective analysis was performed in 126 cases of thoracolumbar fractures. The patients were treated with pedicle screw implantation and the treatment was fol owed up regularly. Ten specimens of fresh calf chest and lumbar spines were obtained. Five specimens served as an experimental group to prepare models of anterior middle column fractures, and these specimens received pedicle screw fixation into the injured vertebra. The other five specimens served as the control group, and did not receive any treatment. In the two groups, along the sagittal axis, axial compression displacement, anterior flexion displacement, posterior extension displacement, torsion angle and pul -out strength were measured and recorded. RESULTS AND CONCLUSION: (1) Clinical trial results: 126 patients were fol owed up for 10-36 months. Compared with pre-treatment, compression ratio of the injured vertebra, the height of intervertebral space, vertebral slip distance, visual analogue score and Cobb angle were significantly improved at 1 month after treatment and during final fol ow-up (al P < 0.05). No significant difference in above indexed was detected between 1 month and final fol ow-up (al P > 0.05). At the end of fol ow-up, Frankel spinal cord injury grade was significantly improved (Z=-4.127, P < 0.05). After operation, three patients suffered from pulmonary infection, four patients affected urinary tract infection, and one patient experienced delayed wound healing. (2) Biomechanical results: axial compression displacement, anterior flexion displacement, posterior extension displacement, torsion angle and pul -out strength were smal er in the experimental group than in the control group (al P > 0.05). (3) Above findings showed that pedicle screw fixation into the injured vertebra for thoracolumbar fractures obtained good long-term effects, and had good biomechanical properties.

Objective The aim of this study was to investigate the possible effect of glycated low density lipoprotein ( LDL ) on the proliferation, differentiation, and expression of low-density lipoprotein receptor-related protein5(LRP5)mRNA,dickkopf-1(DKK1)mRNAinmouseosteoblasts(MC3T3-E1cells). WhethertheWnt signaling pathway was involved in the above process was explored else. Methods Mouse MC3T3-E1 cells were culturedwithvariousconcentrationsofglycatedLDL(glycatedLDLlevelwas2.4%,5.3%,8.7%,13.9%)for24, 48, 72 h. Proliferation of MC3T3-E1 cell was measured by CCK-8, the osteocalcin level in the medium was determined by ELISA and the expression of LRP5 mRNA, DKK1 mRNA was measured by realtime PCR. Results After cells being incubated with 5. 3% of glycated LDL for 24 h, the inhibition of MC3T3-E1 cells was more marked than that in control group(P<0. 01). The higher glycated LDL level, the severer the inhibition. The effect of LDL on the MC3T3-E1 cell proliferation was time- and dose-dependent under certain conditions. Osteocalcin level in cell culture fluid with glycated LDL was lowered significantly compared with control group. Meanwhile, the expression of DKK1 mRNA was increased significantly but expression of LRP5 mRNA decreased (P<0. 01) in MC3T3-E1 cells cultured with 5. 3% of glycated LDL for 24 h. Conclusions Proliferation and differentiation of the osteoblasts in mice can be inhibited significantly by glycated LDL. The possible mechanism in the role played by glycated LDL on the proliferation and differentiation in MC3T3-E1 cells seems to be related to expression of LRP5 mRNA, DKK1 mRNA in the Wnt signaling pathway.