AIM: To evaluate the effects of all-trans retinoic acid (atRA) on the proliferation in cultured mouse cerebral microvascular endothelial cells (bEnd.3) . METHODS: Cultured cells were divided into five groups randomly, one as control group, the other four groups were 10-9, 10-8, 10-7 and 10-6 mol/L group. Effects of atRA on proliferation in bEnd.3 cells were detected by flow cytometry and immunocytochemitry of PCNA and MTT at 24 h, 48 h and 72 h. The effects of atRA (10-6 mol/L group) on the expressions of angiogenic genes in bEnd.3 cells were studied using microarray. RESULTS: The results of MTT and flow cytometry showed that all- trans retinoic acid at concentration of 10-6 mol/L significantly inhibited the proliferation of bEnd.3 cells. Immunocytochemical staining showed the expression of PCNA was markedly decreased in bEnd.3 cells at 24 h after treatment with atRA. Microarray results demonstrated that there were 11 down - regulated angiogenic genes and 2 up - regulated angiogenic genes in 10-6mol/L atRA group. CONCLUSION: All - trans retinoic acid at concentration of 10-6mol/L may significantly inhibit the proliferation of bEnd.3 cells treated for 24 h ire vitro via down-regulation of angiogenic genes and PCNA expression.

[Objective] To explore the bone histomorphology effect of QiangGuYin on neck of femur of ovariectomized rats.[Methods]Ninety femal SD rats,weight 230～280 gram,were rangdonmized into three groups:QiangGuYin group(treatment group),Miacalcic group(contrast group)and control group.All rats were variectomized.After 10 weeks,the osteoporosis model succeed,the treatment group,contrast group were given QiangGuYin,Miacalcic respectively.The treatment group used QiangGuYin everyday,1ml/g.The contrast group used Miacalcic 0.72u/kg,everyday hyodermic.And 3,4.5,6 months after the treatment,kill 10 rats to explore.The neck of femur were cut and stined with Masson Golder Trichrome for bone histomorphomemetric contrast group analysis.[Result]The treatment group ovariectomized rats' in 3m,BV/TV compared with statistical significance;compared with control group in Tb.sp,BV/TV,Tb.Th with statistical significance.After 4.5m treatment group Tb.Th compared with contrast group have statistical significance.Give the medicine 6m,Tb.sp,Tb.Th compared with contrast group all have statistical significance.[Conclusion]Giangguyin can effectively improve the histomorphometry index.The mechanism probably is stimulating osteogenesis and supperssing the cytoactive of osteoclast and trend of the high bone transform.

[Objective]To study whether Cervus and Cucumis Polypeptide(CCP ,Lugua polypeptide) can treat osteoporosis or not. [Methods]Forty 4-month-old female SD rats were randomized into four groups(10 in each group) ,including SHAM,0VX, OVX+CCP1 and OVX+CCP2 groups. Male rats in OVX groups take the ovariectomized surgery. Since the 13th week, rats in groups A and B got 0.9% physiological saline injection once in a day, group C was treated with CCP 0.4mL/(kg·d)injection and group D treated with CCP0.8mL/(kg·d) injection once one day. Twelve weeks later, al rats were sacrificed for obtaining specimen. Took right femurs, measured the bone biomechanical properties. [Results ]After 12 weeks since injection, the biomechanical property of right femurs in group OVX was significantly lower than that of the SHAM group( P<0.01); parameters such as maximum load and elastic load in group OVX+CCP1 and OVX+CCP2 were higher than that of group OVX(P<0.05), but lower than that of group SHAM(P<0.05). The distinction of the above parameters was significant( P<0.05)between group OVX+CCP1 and OVX+CCP2. [Conclusion]1.CCP can significantly im-prove the bone internal biomechanical properties in ovariectomized(OVX) rats, so it has function to treat osteoporosis caused by drop in estrogen level. There are differences of bone biomechanical properties aspects when treated with different dosage of CCP, which shows the tendency of dose-dependent effect. So we can conclude that CCP may have dose-dependent effect.

AIM: To study the cytological characteristics and gene expression of normal cultured bEnd.3, a mouse brain microvascular endothelial cell strain. METHODS: The morphology of bEnd.3 was studied by light and electronic microscopy, its molecular markers were observed by immunocytochemistry. Cell proliferation kinetics and apoptosis were analyzed by flow cytometry and MTT assay, PGE_2 level was measured by ELISA, and expression of the genes that closely related with vascular endothelial functions was studied by gene micro-array. RESULTS: bEnd.3 had morphological characteristics of microvascular endothelial cells (MVEC) growing in a cobblestone pattern, forming tube-like structure or capillary network and having microvilli. Furthermore, bEnd.3 showed positive staining for vW factor and CD34 and secreted high level of PGE_2 (644.55?30.24 ng/L). Gene micro-array analysis showed CD31, CD36, CD105 expression, and other genes closely related to microvascular endothelial functions also expressed at relatively high level. In addition, bEnd.3 responsed sensitively to mitogen such as basic fibroblast growth factor. CONCLUSION: bEnd.3 is a kind of MVEC, and it can be utilized to study the mechanisms of some diseases such as cancers and cardio- cerebral vascular diseases.

AIM: To study the effect and the mechanism of crenulatin, an effective constituent of Chinese traditional medicine, on apoptosis of cerebral microvascular endothelial cells. METHODS: The following terminal concentrations of crenulatin were used in the study: 25 mg/L and 100 mg/L. Apoptosis of mouse cerebral microvascular endothelial cells (bEnd. 3 cell line) was evaluated by flow cytometer, immunocytochemical assay (Fas, Bcl - 2) and Western blotting (caspase - 3) after culture for 24 h. RESULTS: Compared with control group, apoptosis of bEnd. 3 cells in 25 mg/L group was significantly inhibited ( P ＜0.05), but apoptosis in the 100 mg/L group was significantly increased (P ＜ 0.05). In apoptosis inhibited group, the Fas immunocytochemical staining was weaker, the positive cells were significantly decreased ( P ＜ 0.05) and caspase - 3 expression was decreased compared with control group; however, the Bcl - 2 staining was stronger and the positive cells were significantly increased ( P ＜ 0.05). On the other hand, in apoptosis increased group ( 100 mg/L group), the changes were just opposite. CONCLUSIONS: The effect of crenulatin on apoptosis of mouse cerebral microvascular endothelial cells possesses a dual - direction change, inhibitive effect in 25 mg/L and stimulative effect in 100 mg/L group, respectively. The mechanism is related to the alterations of Fas/Bcl - 2 expression and caspase - 3 activity.

] AIM: To study the effect and the mechanism of crenulatin, an effective constituent of Chinese traditional medicine, on apoptosis of cerebral microvascular endothelial cells. METHODS: The following terminal concentrations of crenulatin were used in the study: 25 mg/L and 100 mg/L. Apoptosis of mouse cerebral microvascular endothelial cells (bEnd.3 cell line) was evaluated by flow cytometer, immunocytochemical assay (Fas, Bcl-2) and Western blotting (caspase-3) after culture for 24 h. RESULTS: Compared with control group, apoptosis of bEnd.3 cells in 25 mg/L group was significantly inhibited (P