Promotion of autophagy and the secretion of CXCR4 of human umbilical cord mesenchymal stem cells induced by low-dose immunosuppressive durgs / 中华器官移植杂志

Objective To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on autophagy and the secretion of chemokine receptor CXCR4 induced by low-dose immunosuppressive durgs.Methods Flow cytometry was used to detect the changes of hUC-MSCs surface markers after treatment with low-dose tacrolimus and rapamycin.The effect of treatment with tacrolimus and rapamycin on proliferation of hUC-MSCs was analyzed with WST-1 assay.Regular RT-PCR was applied to analyze the mRNAs expression of ligands such as LC3B,Atg5 and Beclin1 in hUC-MSCs.Western blotting was carried out to detect the expression of LC3B,Atg5,Beclin1 and p-ULK1 in hUC-MSCs after treatment with tacrolimus and rapamycin.The secretion of chemokine receptor CXCR4 in hUC-MSCs was analyzed under the state of autophay by flow cytometry.Results Flow cytometry analysis confirmed low-dose immunosuppressive drugs tacrolimus and rapamycin did not cause changes in hUC-MSCs phenotypes significantly.Low-dose tacrolimus had no cytotoxic effect on hUC-MSCs,while,rapamycin could inhibit the proliferation of hUC-MSCs after 24 h or 48 h,with survival rate being 73.66％ and 68.81％ (P＜0.05) of controls,respectively.Moreover,both tacrolimus and rapamycin could inhibit PI3K/AKt/mTOR signaling pathway to activate hUC-MSCs autophagy,and the related proteins of LC3B,Atg5 and Beclin1 increased significantly and induced the up-regulation of CXCR4 secretion.Conclusion Our results here demonstrated that low-dose tacrolimus and rapamycin induce autophagy in hUC-MSCs and promote the secretion of CXCR4.

Establishment of reference range for thromboelastography in pregnancy / 中国输血杂志

Objective To establish a reference range for the normal value of thromboelastography (TEG) in pregnant females.Methods According to the results of pregnancy and physical examination,166 pregnant females and 64 healthy females without pregnancy were selected as the pregnant group and the non-pregnant control group,respectively.The TEG value and the traditional coagulation index were measured.The TEG parameters of the two groups were compared and analyzed,establishing a reference range for the parameters.We further analyzed the effect of full-term pregnancy on TEG results and the correlation between traditional coagulation index and TEG test results.Results The traditional coagulation index and TEG test results of the pregnant females andthe non-pregnant females were significantly different.According to the results,a new TEG reference range was established:R 3.9-7.5 min,K 1.0-2.4 min,α 57.6°-74.9°,MA 55.7-75.7 mm,LY30 0-0.56％,CI(-0.97)-3.6.Full-term pregnancy had no significant effect on TEG results.In addition to LY30,other parameters of TEG had some correlation with the traditional coagulation index.Conclusions The general TEG reference range does not apply to pregnant females and established TEG normal reference range for pregnant females can be applied for clinical use.

Comparative analysis between HP-083/4 and rational used instrument on results of six items / 国际检验医学杂志

Objective To compare the performance of HP-083/4 and rational used instrument on detecting six items.Methods The rational instruments were used as contrast instrument,HP-083/4 was the verified instrument.A total of 100 blood specimens and 100 urine specimens were collected,and the levels of antistreptolysin O(ASO),hypersensitive C reactive protein (hsCRP),D-di-mer(D-D),glycosylated hemoglobin(HbA1c),rheumatoid factor(RF)and urine microalbumin(mAlb)were detected.The regression equation and correlation coefficient(r)of the two methods were calculated,and the Kappa values(κ)were analyzed to evaluate the performance of HP-083/4.Results There was a good linear correlation (r >0.950)for the two methods in detecting the serum ASO,hsCRP,D-D,HbA1c,RF and mAlb,r were 0.991,0.995,0.970,0.957,0.980 and 0.967 respectively.Besides,they had good concordance(κ>0.6),theκ values were 0.830,0.957,0.601,0.720,0.920 and 0.694 respectively.Conclusion HP-083/4 is effec-tive in detecting ASO,hsCRP,D-D,HbA1c,RF and mAlb,which should be suitable for clinical application.

Research on multiple myeloma cell apoptosis by inhibition of mTORC2 and chaperon pathways / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore apoptosis of multiple myeloma (MM) cells and its mechanism by the combined inhibition of mTORC2 signaling pathway and heat shock protein 90.</p><p><b>METHODS</b>The effects of Rapamycin, 17-AAG and the combination on proliferation of MM cell lines U266 and KM3 were assessed using MTT at different time points (0, 8, 24, 48 hour). Cell apoptosis and cell cycle distribution were measured by flow cytometry. The specific proteins p-AKT (ser473), p-AKT (thr450), p-S6 (S235/236) and AKT were detected by Western blotting.</p><p><b>RESULTS</b>Rapamycin, 17- AAG and the combination suppressed the proliferation of MM cell lines U266 and KM3, especially the combination of Rapamycin and 17-AAG synergistically inhibited the proliferation (P<0.05); Rapamycin induced G1 arrest both at 24 and 48 hours, 17-AAG also induced G1 arrest, especially at 48 hours (P<0.01); Rapamycin, 17-AAG alone decreased the expression of AKT and induced MM cell apoptosis to some extent (P<0.01); Chronic rapamycin treatment inhibited mTORC2; Inhibition of both mTORC2 and chaper on pathways degraded AKT and induced MM cell apoptosis, which was significantly higher than that of any single agent (P<0.01).</p><p><b>CONCLUSION</b>Inhibition of both mTORC2 and chaper on pathways decreased the expression of AKT to induce apoptosis of MM cells in vitro.</p>

Glycogen synthase kinase-3 and tumorigenesis / 国际肿瘤学杂志

It has recently been demonstrated that Glycogen synthase kinase-3β (GSK-3β) plays an important role in tumorigenesis through its regulation of tumor cell growth via a variety of mechanisms. In some tumors, activation of GSK-3β inhibits the growth of tumor cells whereas in other tumors, GSK-3β promotes tumor cells growth, meanwhile, GSK-3β may also regulate sensitivity of chemotherapy drugs to tumor cells,thus GSK-3β is likely to become a novel target for tumor therapy.

Prevention of aminoguanidine against impairment of endothelium-dependent relaxation induced by glycosylated proteins in isolated rat aorta / 中国药理学通报

AIM This study was designed to investigate the effect of aminoguanidine, an inhibitor of the glycosylated proteins formation, on the impairment of endothelium dependent relaxation induced by advanced glycation end products (AGE) in isolated rat thoracic aorta and its possible mechanisms. METHODS Exogenous glycosylated bovine serum albumin (AGE BSA) was prepared. Aortic rings were exposed to AGE BSA for 60 min to induce the impairment of endothelium dependent vasodilatation. In the drug treated groups, aortic rings were incubated with drug for 15 min and then exposed to AGE BSA for another 60 min in the presence of the drug. Vasodilator responses to acetylcholine (ACh) or sodium nitroprusside (SNP) of aortic rings were measured by isometric tension recording after drug treatment. RESULTS AGE BSA significantly inhibited the endothelium dependent relaxation in response to ACh, but did not affect endothelium independent relaxation in response to SNP. Pre incubation of aortic rings with aminoguanidine(50～500 ?mol?L -1 ) for 15 min and co incubation of aortic rings with AGE BSA for another 60 min markedly attenuated the inhibition of endothelium dependenet relaxation induced by AGE BSA in a dose dependent manner. Superoxide diamutase (SOD, 2?10 5 U?L -1 ), a scavenger of superoxide anions, also prevented the inhibition of endothelium dependent relaxation, which is similar to the effect of 500 ?mol?L -1 aminoguanidine. Furthermore, aminoguanidine (500 ?mol?L -1 ) also reversed impairment of endothelium dependent relaxation of rat aortic ring induced by endogenous oxygen free radicals generated by diethyldithiocarbamate (DETC, 10 ?mol?L -1 ) via inhibiting intracellular SOD. CONCLUSION Aminoguanidine can protect rat aortic endothelium against damage due to AGE BSA, and the beneficial effect of aminoguanidine may relate to its antioxidant properties.

Regulatory effect of indoleamine 2,3-dioxygenase on immune system / 中国药理学通报

The regulation of immune response is affected by several factors in order to maintain homeostasis.Indoleamine 2,3-dioxygenase(IDO) is a tryptophan-catabolizing enzyme expressed by different types of APC,including macrophages and dendritic cells.IDO activity leads to regulatory effects on T cells,which are mediated by tryptophan depletion in specific local tissue microenvironments,the production of proapoptotic metabolites,and inhibition of T-cell clone proliferation through regulatory T cells.IDO plays an important role in protecting the allogeneic fetus from rejection,the development of autoimmune diseases,allograft rejection,and cancer.So regulation of IDO activity may offer a novel drug-based strategy to treat immune-related diseases.

Relationship between the elevation of endogenous inhibitor of nitric oxide and metabolic control in streptozotocin-induced diabetic rats / 中国药理学通报

AIM To determine the relationship between the elevation of endogenous inhibitor of nitric oxide synthase (NOS)N G,N G-asymmetric dimethylarginine (ADMA) and metabolic control in diabetic rats. METHODS Diabetes was induced in Sprague-Dawley rats by a single intraperitoneal injection of streptozotocin. At 72 h after injection, half of diabetic rats received insulin treatment for 8 weeks (20 U?kg -1?d -1,ih, bid). Serum levels of ADMA were measured by high-performance liquid chromatography. Thoracic aortic rings from non-diabetic age-matched control, untreated diabetic, and insulin-treated diabetic rats were tethered in isolated organ baths,contracted with 1 ?mol?L -1 phenylephrine, and challenged with either the endothelium-dependent vasodilator acetylcholine or the endothelium-independent vasodilator sodium nitroprusside. Serum concentrations of glucose, glycosylated serum protein, and malondialdehyde, derived from lipid peroxidation were also examined to estimate metabolic control.RESULTS Serum levels of ADMA significantly elevated in untreated diabetic rats compared with control rats. This elevation of ADMA was accompanied by impairment of relaxation response to acetylcholine but not sodium nitroprusside in aortic rings. Chronic insulin treatment not only prevented the elevation of serum ADMA, but also improved the impaired endothelium-dependent relaxation in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialdehyde were significantly increased in parallel with the elevation of ADMA in untreated diabetic rats compared with control rats. These parameters were normalized after diabetic rats received insulin treatment. CONCLUSION These results provide the first evidence that the elevation of endogenous inhibitor of NOS in streptozotocin-induced diabetic rats is close related to metabolic control of the disease.