8 ?mol/L)was markedly restrained. Zearalenone, like estradiol, could markedly increase the proliferation of MCF-7 cells. The effects were time-dependent and dose-dependent. Conclusion: The tested phytoestrogens are biologically active, and they can differently affect the proliferation of human breast cancer cells in vitro. These data suggest that genistein may have preventive and therapeutic applications against breast tumors.

Objective: This study was designed to investigate the molecular mechanisms of proliferation and apoptosis by genistein and zearalenone through regulation of mRNA and protein expression of PCNA, bcl-2 and bax in breast cancer MCF-7 cells. Methods: The cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments, the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran–treated FBS. The cells were harvested and seeded in 6-well culture plates or in 75 ml flacks. After various concentrations of genistein and zearalenone treatments for 72 h, the cells were harvested and mRNA and protein expression of proliferation cell nuclear antigen(PCNA), bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. Results: At the concentration of 75 ?mol/L, GS could significantly down-regulate bcl-2 and PCNA mRNA expression and up-regulate bax mRNA expression, and zearalenone indicated an opposite result. These results were further confirmed by following immunohistochemistry. Conclusion:PCNA, bcl-2 and bax pathway might be involved in cell proliferation and apoptosis events regulated by dietary estrogens genistein and zearalenone in breast cancer MCF-7 cells.

Objective: To characterize the effects of genistein (GS) on tumor-associated gene expression in MCF-7 cells and to explore the mechanisms. Method: MCF-7 cells were treated with GS (75?10-6 mol/L) or with vehicle (0.1% ethanol, EtOH) for 72 h. The cells were collected and total RNA was extracted, marked by two different fluorescence dyes (Cy3 and Cy5) using reverse transcriptional reaction, respectively. The derived cRNA was hybridized to human tumor-associated microarrays. Data were processed using the Scan Array 3000 and ontological analysis was performed using the Ima Gene 3.0 software. Results: At 72 h, 18 transcripts were downregulated compared to EtOH vehicle control by greater than 2- fold and 4 were upregulated by less than 0.5-fold. The predominant ontological groupings were oncogenes, cell proliferation, apoptosis, and estrogen-regulation genes. Conclusion: GS treatment is associated with significant changes in gene expression in several functional categories in MCF-7 cells, and this might account for the role of genistein in prevention of breast cancer.

Objective To investigate the results of comminuted or open tibi al Pilon fractures treated with a joint bridging movable external fixator. Methods From January 2000 to May 2002, fifteen patients with either severely comminuted or open Pilon fractures were treated with self-designed joint bridging movable external fixators. In some patients, the external fixation was combined with li mited open reduction and Kirschner wire internal fixation. Results All patients were followed up. The average period of following up was 10 months (range 6 to 2 2 months). The fractures were externally fixed for averagely 3 months. The inter val between fracture and bony union was averagely 4.5 months. There was no nonun ion occurred. Two delayed unions were further treated with bone grafting and one of them was stabilized with a plate. All the 9 open fractures were infection fr ee, so were the closed injuries. There was no skin sloughs found in all cases bu t minor pin tract infection was found in two cases. Range of motion at last foll ow-up examination was excellent in five patients, good in seven, fair in two, an d poor in one. One patient required ankle arthrodesis for posttraumatic arthriti s. Conclusion The results of the severely comminuted or open tibial Pilon fractu res treated with joint bridging movable external fixators were fairly satisfying . The fixator can firmly stabilize the fracture while ankle movement can be resu med early. The operation is less-invasive, and is with higher union rate and few er complications.

Objective To study the effects of dietary estrogens genistein(GS)and zearalenone(ZEA)on apoptosis in-duced by estrogen depletion in PEO4cells.Methods The monolayer ovarian cancer cell line PEO4cells were cultured in DMEM medium containing10%bovine serum.Before the addition of the testing compounds the cells were washed in phosphate-buffered saline and the medium was displaced with a phenol red-free DMEM medium containing5%dextral charcoal-stripped FBS and the cells were cultured for5days in order to exhaust the estrogen stored in the cells,and then cells were divided into5groups,including solvent control group,estrogen control group,anti-estrogen control group and2experimental groups.After treatment the apoptotic features of the cells were observed by cellular morphology,DNA fragmentation and location and height of cell hypodiploid were indicated by flow cytometry.Results The typical characteristics of apoptosis in PEO4cells were observed after estrogen deletion and then disappeared following exposure of the PEO4cells to32?10 -9 mol/L and96?10 -9 mol/L ZEA for72hrs.32?10 -6 mol/L and96?10 -6 mol/L GS could significantly aggravate apoptosis in PEO4cells.Conclusion Zearalenone is a kind of mycoestrogen that has estrogenic activity to inhibit apoptosis in PEO4cells.Genistein is a kind of phytoestrogen that has anti-estrogen activity(tamoxifen-like)to promote apoptosis in PEO4cells with the high doses range.

Objective To investigate the effect of vitamin D (VD) on macrophage to phagocytize Staphylococcus aureus (SA). Method Macrophoge cell lines RAW264.7 were allocated into 3 groups:control group(C),bacterium group(B),active vitamin D+ bacterium group (VD+B). Cells in the VD+B group were incubated with 10-8mol/L active vitamin D for 24h,then cells in the VD+B group and the B group were cultured with SA for 1h,and phagocytosis rate,mitochondrial membrane potential,[Ca～(2+)]i,reactive oxygen species were determined by flow cytometry (FCM). Results The phagocytizing activity of macrophage in VD+B group was significantly higher than that in B group 1h after infection,At the same time,the mitochondrial member potential and [Ca～(2+)]i of macrophage in VD+B group were distinctly lower than that in B group; but reactive oxygen species of macrophage in the VD+B group was insignificantly different from B group. Conclusion Vitamin D can reinforce the phagocytizing activity of macrophage and inhibit the apoptosis of macrophage after phagocytize Staphylococrcus aureus.

Objective: To explore the mechanisms of the effect of isoflavone in reducing prostate cancer incidence throught studying the effects of isoflavone on apoptosis in PC-3 cell. Methods: Prostate cancer cell PC-3 was grown in RPMI 1640 medium supplemented with 10% bovine serum and 10 000 U/ml of penicillin/streptomycin in an incubator maintained at 5% CO 2 95% air and 100% humidity at 37 ℃. The respective test compound was added in fresh medium and the control cell received only the vehicle (MDSO). Apoptosis of PC-3 cells was analyzed by cell morphology under light microscope, agarose gel electrophoresis and flow cytometry. Results: Exposure of PC-3 cell to 50 ?mol/L GS, 75 ?mol/L DA and 75 ?mol/L GL after 72 h, the cell morphology indicated typical features commonly used to define apoptosis; agarose gel electrophoresis demonstrated laddered electrophoretic profiles of oligonucleosomal DNA fragments indicative of apoptosis, and flow cytometric analysis revealed a hyperdiploid population in the tested cells. Conclusion: Isoflavones could induce apoptosis in prostate cancer cells PC-3 and it may be the main cause of their role in cancer inhibition.

Objective To investigate the changes of serum osteocalcin and bone micro-structure in ovariectomized mice exposed to low-iron environment.Methods Twenty-four 12-week-old female C57BL/6 mice were divided equally into sham operation (SHAM) group,model(OVX) group,and low iron(OVX+DFO) group.In low-iron group,deferoxamine(DFO) was injected 3 times per week for 5 weeks after operation ; the other groups were injected with the same dose of 0.9％ normal saline for 5 weeks.The serum,left femur,uterus were harvested after five weeks of treatment.The serum osteocalcin and ferritin levels were measured by ELISA kit,the weight of the uterus was recorded by analytical balance.A high resolution micro-CT was used to scan the left femur for cortical bone and cancellous bone analysis.Results (1) The serum osteocalcin and serum ferritin levels in low-iron group were significantly lower than those in the other 2 groups (P＜0.01) ; (2) Compared with the sham group and ovx group,there were significant decrease of the BMD、BV/TV and Tb.N,but increase of Tb.Th and Tb.Sp in low-iron group (P＜0.01).Conclusion A certain dose of DFO (30 mg/kg) can decrease the serum ferritin levels as well as the bone formation index in ovariectomized mice.