Objective:To study the effect of Eupolyphaga Sinensis Walker (ESW) on red blood cell immune adherence (RCIA) in mice model with blood-deficiency by Cyclophosphamide , aimingatfurther research on the mechanism of blood stasis syndrome and promoting blood circulation and removing blood stasis.Methods:This model was made in mice by i.p. cyclophosphamide (0.01 g/kg).It has been observed that how the influence of ESW on the function of red cell immune adherence. The serum anticardiolipin antibodies (ACA),ACA-IgG?ACA-IgA and ACA-IgM levels were measured by using enzymelinked immunosorbent assay (ELISA). The mice body weight were weighted before model and after model and glandular weight were weighted. The serum levels of trace element zinc calcium were determined in healthy mice.Results:The results showed there is a decline in garland rate of red blood cell C3b receptors (RBC-C3bRR) in mice when animal model of Blood - deficiency by cyclophosphamide (0.01 g/kg) form, the serum ACA-IgG and ACA-IgA levels were markedly elevated,while mice body weight and spleen thymus weight markedly depressed. ESW (25 g/kg) increased RBC-C3bRR in the mice model with Blood-deficiency,restified the mice model body weight loss by cyclophosphamide, increased spleen thymus weight .ESW (25 g/kg) increased serum zinc calcium level in the normal mice.Conclusion:ESW increases activity of CR 1 of red blood cells and RCIA, inhibits serum ACA-IgG and ACA-IgA.ESW enhances serum zinc calcium level, that can be a good biological pharmacodynamics.

Objective To investigate the effect of salinomycin on cancer stem cell formation of prostate cancer cell line DU145 and its possible mechanisms,providing theoretical basis for the clinical application of salino-mycin. Methods (1)DU145 cells were treated with salinomycin. The percentage of ALDH+cells,which was used as the marker of cancer stem cells,was detected by flow cytometry.(2)After treated with salmonin,DU145 cells were subjected to Western-Blot analysis for the expression of mTORsignal pathway-related proteins such as p-70s6k, p-p70s6,p-s6 and so on. 3)DU145 cells were treated with salinomycin combined with mTOR signal pathway inhibi-tor rapamycin,and the ALDH+cancer stem cells were detected using flow cytometer. Results (1)Salmonomycin significantly inhibited ALDH-positive cancer stem cells in DU145cell line(inhibition rate in 77.8%),which was twice as high as that of traditional anticancer drug paclitaxel(which has a inhibition rate of 38.64%). This results suggesting that salinomycin would have the effect of inhibiting cancer stem cells. (2)The expression ofm-TOR p-70s6k,p-p70s6 and p-s6 in mTOR signaling pathway was inhibited by salinomycin in a time-dependent and dose-dependent manner,suggesting that salinomycin would inhibite mTOR signaling pathway.(3)Salinomycin combined with rapamycin can decrease the proportion of ALDH-positive DU145 cancer stem cells(inhibition rate in 77.95%), suggesting that salinomycin may inhibit ALDH-positive DU145 stem cells through the mTOR signaling pathway. Conclusion Salinomycin may play an important role in inhibiting cancer stem cells by inhibiting mTOR pathway signaling.

Amino Acid Substitution ; Exocytosis ; HEK293 Cells ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Membrane Proteins ; chemistry ; metabolism ; Microscopy, Confocal ; Protein Binding ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics ; Sirolimus ; analogs & derivatives ; chemistry ; metabolism ; Tacrolimus Binding Proteins ; chemistry ; genetics ; metabolism

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.