Objective This study was designed to explore the effects of phytoestrogen genistein on proliferation of post-menopausal osteoblast and the likely molecular mechanisms. Methods Osteoblast cultures were prepared from the upper femur of postmenopausal patients. Osteoblast proliferation was determined by Methyl Thiazolyl Tetrazolium(MTT)assay and cell cycle distribution by cytometry. The protein expressions of cyclin D and E were examined using Western-blot. Results Genistein (10~(-8) to 10~(-6) mol/L) stimulated cell viability of postmenopausal osteoblasts and increased the distribution ratio of the cells at the G2/M and S phases (P

Objective: To explore the effects of zinc on bone development in vitro. Methods: Mice of 16 gestation were killed by snapping cervical vertebra, and the front limbs were cut off and cultured in basic medium. Which contained 20 ?mol/L Zn 2+ (control group). In Zn deprived group, the basic medium was added TPEN to final concentration 20 ?mol/L. In other three groups, the basic medium was added ZnSO 4 to final concentration of Zn 2+ at 45, 70, and 120 ?mol/L respectively. Results: 1. Compared with control limb(uncultivated),the test limb(cultivated for 6 d)showed longer bone length and higher bone density.Histological analysis showed active bone cell differentiation, proliferation, increased bone trabecula.2.Compared with control group, the Zn-deprived and Zn 2+ 120 ?mol/L group expressed decreased osteocalcin (OC) and 45 Ca content and lower activity of AKP; Zn 2+ 45 ?mol/L and 70 ?mol/L group presented increased OC and 45 Ca content and higher activity of AKP.3.Compared with the control group, Zn-deprived and Zn 2+ 120?mol/L group showed shorter bone length and lower bone density; Zn 2+ 45 ?mol/L and 70?mol/L group showed longer bone length and higher bone density. Conclusion: Zinc is involved in bone metabolism and growth; both zinc-deficiency and zinc-excess can alter bone growth and normal metabolism.

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 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 effects of genistein (Gen) on apoptosis in human osteoblast. Methods:Human periosteum was maintained in DMEM medium containing 10% newborn calf serum through four passages to isolate and proliferate osteoblast. Then, cells were maintained in phenol red-free DMEM supplemented with 5% charcoal-stripped FCS for 4 d before addition of treatments. Flow cytometer and DNA fragment were used to explore apoptosis in osteoblast, and immunohistochemistry was used to study protein expression of Bax, Bcl-2 and caspase-3. Results:Estrogen depletion could induce apoptosis in human osteoblast. With the similarity of 10-8 mol/L E2, at the conentrations of 10-6 mol/L and 10-5 mol/L, Gen could inhibit apoptosis induced by estrogen depletion. Compared with vehicle control, Gen significantly decreased protein expression of Bax and caspase-3 and increased protein expression of Bcl-2. Conclusion:Genistein could inhibit apoptosis induced by estrogen depletion in human osteoblast.

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: 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.

0.05, respectively), but those to noradrenaline (NA: 0.01?mol/L), histamine (His: 3?mol/L), and 5-hydroxytryptamine (5-HT: 0.1?mol/L), significantly reduced (P0.05, respectively). Under atropine or Ani pretreatment, the NA-, His- and 5-HT-elicited contractions in endothelium-denuded aorta were similar to those in endothelium-intact aorta. Conclusion Atropine and Ani can inhibit receptors-mediated constrictions of rabbit aortic vascular smooth muscle cells; the actions are endothelia independent.

AIM: To observe the changes of interleukin-6(IL-6), IL-8 and tumor necrosis factor-?(TNF-?) in serum and lung at different time, and the effects of anisodamine (654-2) treatment in rats with oleic acid-induced ARDS. METHODS: The ARDS model induced by intravenous injection of oleic acid in the rat was used and levels of IL-6, IL-8, TNF-? in serum and lung tissue supernatant were measured using enzyme linked immunosorbent assay (ELISA). RESULTS: Levels of serum and lung tissue IL-6, IL-8, TNF-? in oleic acid type ARDS 4 h group were increased significantly. These cytokines in oleic acid type ARDS 8 h group were lower than that of ARDS 4 h group, but serum IL-6, TNF-? and lung tissue IL-6 were still higher than that of control group . In oleic acid type ARDS 16 h group, serum IL-6, TNF-? were lower than that of the ARDS 8 h group and serum TNF-? and lung tissue IL-6 were higher than that of control group. After 654-2 treatment, the levels of serum and lung tissue IL-6, TNF-? were decreased significantly. CONCLUSION: IL-6, IL-8 and TNF-? might play important roles in the oleic acid-induced ARDS in the rat. 654-2 might alleviate ARDS by inhibiting excess production of IL-6 and TNF-?.

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.