Study on relationship between BMP9-induced osteogenicdifferentiation and COX-2/PI3K/Akt in stem cells / 中国药理学通报

Aim To investigated the possible effect of COX-2 on the BMP9-induced activation of PI3K/Akt signal in progenitor cells.Methods The activity of alkaline phosphatase(ALP) was measured using histochemical staining or chemiluminescence.The mRNA level of ALP was determined using real-time PCR assay.The protein levels of osteopontin(OPN), osteocalcin(OCN), COX-2, Akt1/2 and phosphorylated Akt1/2 were detected by Western blot.The mRNA level of COX-2 was assayed with RT-PCR, and the mineralization was measured with Alizarin Red staining.Results The ALP activity was apparently increased by BMP9 in C2C12 cells, as well as the protein level of OPN and OCN.The mineralization was also markedly induced by BMP9 in C2C12 cells.BMP9 increased the level of phosphorylated Akt1/2 greatly, although no substantial effect was observed on total protein level of Akt1/2.The BMP9-induced ALP activity was dramatically decreased by the inhibitor of PI3K.The mRNA and protein level of COX-2 were both increased by BMP9 in C2C12cells, and the BMP9-induced ALP activity and mineralization were greatly attenuated by the inhibitor of COX-2.The BMP9-induced phosphorylation level of Akt1/2 was increased by the exogenous expression of COX-2, but decreased by the inhibitor of COX-2.Conclusion Activation of PI3K/Akt signaling may be a critical event in BMP9-induced osteogenic differentiation, and this process may be mediated by the BMP9-upregulated COX-2 in stem cells at least.

Study on relationship between anti-proliferation effect of resveratrol and resveratrol-induced bone morphogenetic protein 9 and its receptors in colon cancer cells / 中国药理学通报

Aim To study the anti-proliferation effect of resveratrol (Res)and the role of Res-induced bone morphorgenetic protein 9 (BMP9 )in this process in colon cancer cells.Methods Crystal violet staining and flow cytomtry were introduced to assay the anti-proliferation effect of Res in LoVo cells.The effect of Res on apoptosis in LoVo cells was also detected with flow cytometry.Then,RT-PCR and Western blot assay were employed to unveil the effect of Res on the ex-pression of BMP9 .The effect of BMP9 on the anti-pro-liferation of Res in LoVo cells was analyzed with crystal violet staining and flow cytometry too.Finally,the effect of Res on the expression of ALK2 and ALK3 was assayed with RT-PCR,and the inhibitor of ALK2 and ALK3 was used to figure out the possible mechanism of BMP9 on Res-induced proliferation inhibition in LoVo cells.Results Res apparently inhibited the prolifera-tion,arrested the cell cycle at S phase in LoVo and in-creased the percentage of apopotic cells in LoVo cells. Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous ex-pressed-BMP9 enhanced the anti-proliferation and ap-optosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res.Al-though Res had no apparent effect on increasing the phosphorylation of Smad1/5/8,it increased the ex-pression of ALK2 and ALK3 .Inhibition of ALK2 and ALK3 decreased the anti-proliferation effect of Res partly in LoVo cells.Conclusion Res is potent to in-hibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.

Anti-proliferation effect of resveratrol and p38 MAPK in human colon cancer cells / 中国药理学通报

Aim To investigate the relationship be-tween the anti-proliferation effect of resveratrol ( Res ) and p38 MAPK in colon cancer cells .Methods Crys-tal violet staining , Western blot and flow cytometry were employed to analyze the effect of Res on the pro-liferation in LoVo cells.Western blot assay was used to detect the effect of Res on the apoptosis of LoVo cells and the phosphorylation of p 38 MAPK.Crystal violet staining and Western blot assay were used to analyze whether p38 MAPK was involved in the Res-induced proliferation inhibition and apoptosis in LoVo cells .Re-sults Res inhibited the proliferation , arrested cell cy-cle at S phase , and increased the protein level of PC-NA in LoVo cells apparently .Res increased the level of Bad in LoVo cells, but decreased the level of Bcl-2. Although Res exerted no substantial effects on total lev-el of p38 MAPK, it markedly increased the phospho-rylation level of p38 MAPK in LoVo cells.p38 MAPK inhibitor promoted the proliferation , and decreased the anti-proliferation effect of Res on LoVo cells .Moreo-ver , the effects of Res on the level of Bcl-2 and Bad were both reduced by the p 38 MAPK inhibitor .Con-clusions Res can inhibit the proliferation of LoVo cells, which may be partly mediated by promoting the phosphorylation of p38 MAPK.

LONGEVITY AND PERIODICITY OF MICROFILARIAE OF BRVGIA MALAYI TRANSFUSED TO VARIOUS STRAINS OF LABORATORY MICE / 中国寄生虫学与寄生虫病杂志

Microfilariae (mff) obtained from the peritoneal cavities of infected jirds (Meriones unguiculatus) were injected intravenously into 6 mouse strains: SMMC/B, BALB/cCR, LACA, ICR/JCL, 615 and Kunming strain. The mff in the peripheral blood of the 6 mouse strains in the above sequence were detectable within 1-150, 24-80, 30-60, 3-96, 12-45 and 45-60 days after inoculation of 2 ? 105 mff per mouse. The survival period of mff was longer in SMMC/B mice than in mice of the other strains, and the density of mff in blood was also higher in the former than in the latter. Out of 18 SMMC/B mice 13 remained microfilaria-positive on the 60th day after inoculation. The duration and level of microfilaraemia were proportional to the dose of parasites injected Microfilariae disappeared from the peripheral blood of all mice 3 days after injection of 1 ?104 mff, but were still detectable 60 days after injection of 2 ? 105 mff. In addition, mff disappeared much faster from the peripheral blood of SMMC/B mice after the second inoculation than after the first one. Although the mff had already disappeared from the peripheral blood of the infected and reinfected mice for about 1 to 2 months, they could still be found in the internal organs, mostly in the small blood vessels of the lungs (about 90%). The mff maintained a nocturnal sub-periodicity in the recipient mice, similar to those observed in the donor jirds, with a peak density hour between 2:28 and 5: 48 a.m.The results show that this mouse model might be a simple and useful system in which various factors controlling the fate of mff can be studied independently.

Efficacy of Mebendazole Composite Cream in Treatment of Trichiniasis in Mice by Skin Administration / 第二军医大学学报

Male mice weighting 25-32g were infected with larvae of Trichinella spiralis. Skin administrations of mebendazole composite cream were carried out on the mice in 3 different experiment groups. It was found that there were the reduction rate of worms at 97.3%, 100% and 99. 7% in the adult worm group (50 mg/kg x 1 d), in the invasive larvae group (25 mg/kg x 7 d) and in the encysted larvae group (25 mg/kg x 7 d), respectively. These results indicate that mebendazole composite cream is very effective against both intestinal and muscular phases of T. spiralis.