RESUMO
Objective To deplore the immunoregulatory function changes of mesenchymal stem cells(MSCs)from multiple myeloma(MM)patients and its effects on the pathogenesis of myeloma bone disease.Methods MSCs from MM patients and normal controls were isolated and the immunophenotype was detected.Real-time PCR was performed to detect the expressions of TGF-β1,TGF-β2,TGF-β3,IL-6,IL3,TNF-α,FasL and RANKL of MSCs.Transwell coculture systems were performed between MSCs and T cells.Lymphocyte proliferative assay was employed to detect the effect of MSCs on T cell proliferation.The effect of MSCs on T cell cycle and T cell activation markers CD25 and CD69 expression were analyzed by flow cytometry.Cleaved caspase 3 protein by western blot and hoechst 33258 staining were employed to detect the apotosis of T cells.Influence of T cells on the osteogenesis potential of MSCs were detected by Von kossa stain,real-time PCR and Western blot.Results MSCs from both MM patients and normal controls possessed similar morphology and immunophenotypes.MM derived MSCs exhibited increased expressions of TGF-β1,IL-6,IL-3,TNF-α and RANKL and decreased expression of TGF-β2,TGF-β3 and FasL.The inhibitory effect of MM derived MSCs on T cell proliferative ability was attenuated compared to control MSCs.MSCs from normal controls silence more T cells in Go/G1 phase than those from MM patients.The daupening effect of MM derived MSCs on activation-induced T apoptosis seemed to be enhanced.Expression of T cell activation markers were significantly inhibited by MSCs from normal controls.Both T cells cocultured with MM deprived MSCs and T cells directly from MM patients inhibited osteogenesis potential of MSCs from normal controls.Conclusion MSCs from MM patients showed impaired immunoregulatory capability on T cells.The activated T cells,in turn,inhibited the osteogenesis potential of MSCs.This may participate in the pathogenesis of myeloma bone disease.
RESUMO
Objective To analyze the genetic polymorphism of three short tandem repeat (STR) double loci,namely,six STR loci in the Chinese population of Han nationality in Jilin district and to obtain the corresponding genetic data.Methods DNA was extracted from the blood cells of one hundred and three unrelated individuals of Han nationality in Jilin district;Six loci were divided into three groups which were groupⅠ(TPOX+CSF1PO),groupⅡ(D3S1358+D13S317) and groupⅢ(D5S818+D19S400).The extracted DNA were amplified with PCR multiplexing method.The PCR products were analyzed by non-reduced PAGE,followed by silver staining.The allele frequency distributions of six STR loci were studied by statistical procedures.The heterozygosity (H),discrimination power (DP),probability of exclusion (PE) and polymorphic information content (PIC) were calculated.Results The allele frequencies of six loci were obtained.The genotypes distributions of the loci were consistent with Hardy-Weinberg equilibrium and had higher heterozygosities and PIC.The H,DP,PE and PIC were 0.7511-0.8376,0.8273-0.9247,0.5167-0.6718 and 0.7100-0.8195 respectively.Conclusions The three STR double loci exhibited higher polymorphism and were better genetic markers.The data on the allele frequencies of these six STR loci may be used in individual identification,paternity testing and in other population genetic researches.