Objective To study the effect of silencing Nek2 via RNAi on cell cycle of ovarian cancer SKOV 3 cells and the re‐lated molecular mechanism .Methods The Nek2‐siRNA was transfected into the ovarian cancer SKOV3 cells .The change of cell cy‐cle of SKOV3 cells at 48 h after transfection was examined by the flow cytometry technique ;Western blot assay was used to deter‐mine the change of level of the cell cycle related factors cyclinB1 ,CDK1 ,P27 and the phosphorylation level of the ERK1/2 after Nek2‐siRNA transfection for 48 h .Results The flow cytometry detection results showed that the proportion of the cells in G 2/M stage in the blank control group ,negative control group and RNAi group was 13 .72% ,12 .27% and 1 .56% respectively .Compared with the control group ,the number of the cells in G2/M stage in the transfected group was reduced obviously (P< 0 .05) .The Western blot detection results showed that compared with the control group ,the expression of cyclinB1 and CDK1 protein in SK‐OV3 cells was significantly reduced ,the expression of P27 was increased after silencing Nek2 and the phosphorylation level of ERK1/2 in SKOV3 cells was significantly reduced after silencing Nek2 gene(P< 0 .05) .Conclusion Silencing Nek2 gene might block the ovarian cancer cell line SKOV 3 initiating mitosis ,thus inhibit their proliferation .

BACKGROUND: Clinical studies demonstrate that the durable stability of the prosthesis depends on a close geometric fit between the prosthesis and femoral medullary cavity.OBJECTIVE: To study the law of the parameters of medullary canal section shape in proximal femur DESIGN: Repeated measurement observation.SETTING: Department of Orthopaedics, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University MATERIALS: Ten femoral samples without any damage was obtained from corpse in the Staff Room of Anatomy of the Shanghai Second Medical University METHODS: The experiment was carried out in the Department of Or thopaedics, Ninth People's Hospital between January 2000 and March 2000. Section morphology of medullary cavity of proximal femur was dealt with image-processing, and conus curve fitted parameter mathematical method was proposed; and at the same time, section of medullary cavity of proximal femur of 10 patients was measured manually.MAIN OUTCOME MEASURES: Computer-aided method and manual method were applied to measure the coordination at the end of extrude and the coordinate of link joint RESULTS: 10 femoral samples entered the stage of result analysis. Com puter was used to measure the coordination of the end of extrude of the section medullary cavity of proximal femur (X,Y) and the coordination of the connection point (X,Y).There was no significant difference of the measuring results between computer-aided method and manual CONCLUSION: Computer-aided imaging-processing method not only reduces the error but also can be completed by computer automatically in measuring section morphology of medually cavity in proximal femur. It is suitable for a variety of morphology measurement.

Objective To explore the effects of alendronate on adipogenic differentiation of bone marrow stremal cells (BMSCs) and the role of mitogen-activated protein kinases (MAPK) signal pathway in this process. Methods BMSCs were derived from 9-month-old ovariectomized SD rats and exposed to 0.01, 0.1, 1, 10 μmol/L of alendronate for 2 weeks. The number of BMSCs was counted under light microscope after oil red O staining. The expression of peroxisome proliferators activated receptor-γ, 2 (PPAR-γ2) was measured by RT-PCR. The effect of alendronate on MAPK signal pathway was detected by Western blot. Results After two weeks of induction of BMSCs by alendronate, BMSCs with positive oil red O staining significantly decreased as the increase of the concentration of alendronate (P <0.01), so did the expression of PPAR-γ2. The expression level of PPAR 2 increased when exposing BMSCs to ERK1/2 or JNK specific inhibitors, PD98059 and SP600125 for two weeks. However, the expression level of PPAR 2 decreased when exposing BMSCs to SB203580 (an inhibitor of p38) for two weeks. Condusion Alendronate can inhibit adipogenic differentiation of BMSCs derived from ovariectomized rats in a doso-dependent manner by activating ERK1/2 and JNK rather than p38.

Objective To assess the fixation efficiency and biocompatibility of the absorbable high strength poly L lactide (SC PLLA) screws which were designed and made by us through a new material reinforcement process, solid state compression (SC), by investigating the healing and tissue reaction of the rabbits which were given femoral osteotomy and fixed with the screws. Method The right femoral condyles of 12 New Zealand rabbits were first cut off and then fixed with a SC PLLA screw. They were sacrificed at 4, 8, 12, and 36 weeks postoperatively to exam their femurs radiologically and histologically. Results At all the sites of osteotomy was seen obvious formation of bony callus. There were no significant displacements at all the operative sides. Histologically, bony union showed at the site of osteotomy and direct connection between the bone and the implants was observed. Conclusion SC PLLA screws can provide sufficient internal fixation for the load bearing cancellous bone, and possess excellent biocompatibility.

ObjectiveTo study the influence of stress－ relaxation plate on disorganization andrepair of the cortex under the plate. MethodsA washer made of viscoelastic polyethylene was placedbetween the screw and the screw hole of a conventional stainless rigid plate(RP) to form a stress－ relax-ation plate(SRP). Both SRP and RP were applied to the osteotomized New Zealand rabbit tibia, thefracture healing process of either fixing with SRP or RP(control) was put under comparative study bypolarized light microscopy, in situ hybridization of collagen mRNA and immunohistochemistry tech-nique from 2 to 36 weeks postoperatively. ResultsThe study of plated bone remodeling showed thatthe degree of cortex osteoporosis beneath the plate was similar between the SRP and RP group 12 － weekpostoperatively. The organization of the bone structure in the SRP group happened later and milder thanthat of the RP group, and the repair process began 12 weeks after implantation. So the resorption cavi-ties became smaller and the structure of collagen fibers became well oriented along, and the osteoblastslying on the surface of resorption cavities expressed and synthesized type Ⅰ collagen 8 to 12 weeks afterimplantation. The changes above increased significantly in most cavities by 36 weeks. No expression andsynthesis of any kind of collagen could be observed during 12 to 36 weeks after implantation in the RPgroup. ConclusionWithout removal of the bone plate, the SRP fixation not only reduces the degreeof plated bone osteoporosis, but also makes the disorganized bone structure return to normal by the ex-pression and synthesis of type Ⅰ collagen mRNA of osteoblasts lying on the surface of resorption cavities.

Objective　To explore the influence of stress-relaxation plate(SRP) fixation on the remodeling of cortex under plate. Methods　Twenty-eight New Zealand rabbits were used in this study, the bilateral tibia were osteotomized in the middle and fixed with SRP (experimental group) and rigid plate (control group) respectively. The scanning electron microscopy was used to observe the bone remodeling process from 2 to 48 weeks after operation. Results　There was cortex osteoporosis beneath plate in different degree in both experimental and control groups before 8 weeks, it showed as the disorganization of collagen fiber structure and formation of resorption cavities. In comparison, the osteoporosis degree in experimental group showed milder than that of the control group. After 12 weeks, the resorption cavities became smaller, and the structure of collagen fibers became regular with the alignment parallel to the long axis of cortex. In contrast to the experimental group, the bone osteoporosis under plate of control group exacerbated continuously. Conclusion　Without removal of the bone plate, SRP fixation not only reduce the degree of plated bone osteoporosis, but also make the osteoporosic bone return to normal.

Objective To investigate fluoride-induced inflammation and nuclear factor-κB (NF-κB) signaling pathway in cultured human acute monocytic leukemia cells (THP-1).Methods In vitro cultured THP-1 cells were used as a model of microglia.After cultured with different concentrations of [0 (negative control group),10,50,100,500,1 000 and 5 000 μmol/L] sodium fluoride (NaF) for 48 h,the survival of cells was detected by CCK8.THP-1 cells were divided into 3 groups:control group,low dose and high dose fluoride groups according to the results of CCK8 assay,and then treated with different concentrations of sodium fluoride (0,500,5 000 μmol/L) for 48 h,concentrations of inflammatory cytokines,such as Interleukin-lβ (IL-1β) and tumor necrosis factor-α (TNF-α) were measured by enzyme linked immunosorbent assay (ELISA) in THP-1 mononuclear cell culture medium.The protein levels of IκBα,phospho-NF-κB p65 and phospho-IκB-α were detected by Western blotting.Results THP-1 cells were treated with different concentrations of sodium fluoride (500,1 000,5 000 μ mol/L) for 48 h.Fluoride group THP-1 cell survival rate [(73.21 ± 3.67)％,(31.40 ± 4.56)％,(0.40 ± 0.24)％] was lower than that of the negative control group [(100.00 ± 0.00)％,all P ＜ 0.01].Compared to the control groups [(0.36 ± 0.07),(31.07 ± 0.81)ng/L],significant increases of the inflammatory cytokines IL-1β [(1.42 ± 0.79),(19.47 ± 2.90)ng/L] and TNF-α [(61.06 ± 2.20),(172.72 ± 2.29)ng/L] were detected in culture medium in low-fluoride and high fluoride groups,respectively.Interestingly,compared to the control groups [(100.00 ± 5.48)％,(100.00 ± 14.82)％],significant increases of phospho-NF-κB p65 [(113.71 ± 8.99)％,(134.74 ± 1.93)％] and phospho-IκB-α [(152.61 ± 14.16)％,(176.91 ± 7.95)％] were observed in both low-fluoride and high fluoride groups.Meanwhile,the protein level of IκBα in high fluoride group [(63.53 ± 9.67)％] was significantly lower than that of the control group [(100.00 ± 10.99)％,P ＜ 0.01].Furthermore,significant positive correlation was detected between increased IL-1β,TNF-α and phospho-NF-κB p65 (r =0.74,0.72,all P ＜ 0.05).Conclusions Excessive fluoride can induce microglial cells to release inflammatory cytokines and activate nuclear factor-κB signaling pathway.The release of inflammatory cytokines and activation of the signaling pathway may be one of the mechanisms of the damage of the central nervous system caused by sodium fluoride.

Objective To evaluate effectiveness of bone marrow derived mesenchymal stem cells(BMSCs) transduced by adenovirus mediated human bone morphogenetic protein-2 (Adv-hBMP-2) gene in the repair of diaphysic segmental bone defects of goats. Methods The tibial bone defects (2.1 cm) of 22 goats, who weighted from 18.1 kg to 29.5 kg, were established and divided into 4 groups: groupⅠ, Adv-hBMP-2 transduced BMSCs group (n=8); groupⅡ, adenovirus mediated ?-galactosidase(Adv-?gal)gene transduced BMSCs group (n=6); groupⅢ, non-transduced BMSCs group (n=6); groupⅣ, untreated group (n=2). Roentgenography, histomorphometrical analysis and biomechanical measurement were studied at various times. Results Roentgenography showed more callus in the bone defect site of group Ⅰ. At 24th week after implantation, the healing rates of group Ⅰ,Ⅱ,Ⅲ, and Ⅳ were 6/7, 1/5, 2/5, and 0/1 respectively. There was statistic significant difference of X-ray grading between groupⅠand groupⅡor Ⅲ. Histomorphometrical analysis showed much more total new trabecular bone area (TBA) of groupⅠthan that of other groups. Biomechanical strength of groupⅠwas superior to that of groupⅡand Ⅲ. Conclusion Adenovirus mediated BMP-2 gene transferred BMSCs can repair segmental bone defect in goat without osteoconductive scaffold.

Objective To investigate the rejection mechanisms of bone grafts by comparing the immune reaction of xenografts versus allografts, and the influence of IL-4 and IL-10 on immune responses. Methods Twenty C57BL/6 mice and 1 New Zealand rabbit were used as transplant donors for allografts and xenografts, respectively. One hundred BALB/c mice were used as transplant recipients and randomly divided into 5 groups. Using a model of the muscle pouch for implantation, the immune reactions of bone allografts and bone xenografts were studied through observing the lymphocyte responses of the stimulating index of lymphocytes from the mixed lymphocyte culture (MLC), subsets of lymphocyte, cytokinetic and histological findings in 1, 2, 4 and 6 weeks after implantation respectively. Results Xenograft rejection was rapid (1 week) and stronger than allograft rejection (P

Objective To evaluate osteogenetic effectiveness of hyaluronic acid (HA) mixed with adenovirus mediated human bone morphogenetic protein-2 gene(Adv-hBMP-2) transfected bone marrow derived mesenchymal stem cells(BMSCs) of rabbits in the repair of radial shaft defects of rabbits. Methods The BMSCs were collected from bone marrow of rabbits, cultured and transfected by Adv-hBMP-2 in vitro. The radial shaft defects (1.5 cm) models of 20 rabbits (40 sides) were created and then respectively injected the cells and/or materials according to the following four groups (n=10): Ⅰ, Adv-hBMP-2 transfected BMSCs+HA; Ⅱ, Adv-hBMP-2 transfected BMSCs; Ⅲ, HA; Ⅳ, control. Results The bone defects of 6 out of 8 sides in the group Ⅰ were repaired completely and partial bone marrow cavities were revasculized. Only 3 sides were repaired completely in the group Ⅱ and no side was repaired completely in the groups Ⅲ and control. There were statistic significance between each groups in X-ray scores. In the groups ofⅠand Ⅱ, much woven callus formed in the sites of the bone defects in the 4-8th week after injection. Partial bone marrow cavities were revasculized in the 12th week after injection; while in the groups of HA and control, the sites of bone defects were mainly full of fibrous tissue and no healing happened. Both ends of the bone defects were hardened. There was significant difference statistically in the new trabecular bone area in the groups ofⅠand Ⅱ. The maximum vertical loading and elastic modulus were significant differenct in the group ⅠandⅡ. The ratios of average maximum vertical loading/normal value of radial shaft in the groups ofⅠandⅡwere 75.86% and 45.45%, respectively. Conclusion The tissue-engineered bone made by HA mixed with Adv-hBMP-2 transfected BMSCs can repair the bone defects in the radial shaft of rabbits. HA is a safe, effective carrier for tissue engineering.