[mTOR signaling pathway-mediated autophagy involved in inhibition of osteoblast differentiation induced by cadmium in human bone marrow mesenchymal stem cells].

Objective: To investigate the role of autophagy mediated by mTOR signaling pathway in the inhibition of osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) induced by cadmium. Methods: HBMSCs were divided into 0, 2.5 or 5.0 µmol/L groups according to the exposure dose of cadmium chloride (CdCl2), and each group was treated for 1 day, 4 days and (or) 7 days. The ALP activity and mRNA and protein expression levels of osteogenesis markers (ALP, RUNX2 and OSTERIX), autophagy-related proteins (LC3 and Beclin-1) and mTOR signaling pathway related proteins (mTOR, p-mTOR and p-p70S6K) expression, alkaline phosphatase staining and alizarin red staining were detected. MHY 1485 was selected as the signaling pathway activator. The control group, CdCl2 group (5.0 µmol/L), MHY 1485 group and CdCl2+MHY 1485 combined treatment group were set. After 7 days of treatment, the expression levels of autophagy related proteins and mTOR signaling pathway related proteins of hBMSCs in each group were detected. Results: There was no significant difference in ALP activity between 0, 2.5 and 5.0 µmol/L groups on day 1 and 4 (P>0.05); On day 7, compared with the 0 µmol/L group, the ALP activity, expression of osteogenic markers (ALP, RUNX2, OSTERIX) and mTOR signaling pathway related proteins (mTOR, p-mTOR, p-p70S6K) expression decreased in the 2.5 and 5.0 µmol/L group (P<0.05). Compared with the 0 µmol/L group, the staining of the 2.5 and 5.0 µmol/L groups became lighter, and the formation of ALP and mineralized nodules was reduced. Compared with the CdCl2 group, the autophagy related protein expression in the CdCl2+MHY 1485 combined treatment group decreased, and the mTOR signaling pathway related protein expression increased. The difference was statistically significant (P<0.05). Conclusion: The inhibition of osteogenic differentiation of hBMSCs by cadmium may be related to autophagy mediated by mTOR signaling pathway.

[Pollution characteristics and health risk assessment of heavy metals in PM(2.5) in Lanzhou].

Objective: To understand the pollution characteristics and assess the pollution health risks of heavy metals in atmospheric PM(2.5) in Lanzhou. Methods: According to the regional characteristics of air pollution and industrial distribution characteristics in Lanzhou, atmospheric PM(2.5) was sampled monthly in Chengguan and Xigu Districts from January, 2015 to December, 2016. Detected the concentration of PM(2.5) and 12 kinds of elements (Sb, Al, As, Be, Cd, Cr, Hg, Pb, Mn, Ni, Se and Tl) by weighing method and inductively coupled plasma mass spectrometry. Enrichment factor and geo-accumulation index were used to describe the pollution characteristics, while health risk assessment was conducted using the recommended United States Environmental Protection Agency (USA EPA) model. The health risks of non-carcinogens were evaluated by non-cancer hazard quotient (HQ), the non-carcinogenic risk was considered to be negligible when HQ<1, HQ>1 meant a health risk. With a single contaminant cancer Risk value to evaluate the health risks of carcinogens, when the Risk value between 10(-6) to10(-4) as an acceptable level. Results: The daily average concentrations of PM(2.5) was 83.0 µg/m(3), 77.0 µg/m(3) in Chengguan and Xigu Districts, respectively, during the sampling periods, and the concentration of PM(2.5) in winter/spring was higher than summer/fall in both districts. The concentration of Al in PM(2.5) was the highest and other elements in descending order: Pb, Mn, As, Sb/Cd, Tl in both districts. Enrichment factor results showed that Al and Mn were mainly affected by natural factors, the rest of five elements were all typical man-made pollution elements and according to geo-accumulation index pollution level of Cd was the strongest in the winter. The results of health risk assessment showed that Mn had the highest non-cancer risks (HQ>1) and affected the health of the children seriously. HQ reached up to 2.44 and 1.79 in Chengguan and Xigu Districts, respectively. Pb, As, Sb, Cd had slight health impact (HQ<1), could be negligible. The cancer risks range of As, Cr were 6.33×10(-6) to 6.46×10(-5) between the acceptable level of risk (10(-6) to 10(-4)), which indicated that As and Cd had potential cancer-risks. Conclusions: The pollution level of atmospheric PM(2.5) and the heavy metals in it was still grim;the non-cancer risks caused by multiple metals on children deserved attention. Although the cancer risks of As and Cd were between the acceptable level of risk, the potential cancer risk still shall not be ignored.

Expression of human beta E-globin gene with erythroid enhancer in transgenic mice.

The human beta E-globin gene from HbE (beta 26 Glu-->Lys, G-->A) homozygote, a common hemoglobinopathy in China, was injected into the pronuclei of fertilized mouse eggs to prepare transgenic mice. A transgenic mouse bearing 16 copies of construct 5'.HS2 beta E has characteristics of typical transgenic mice. It has been verified that the presence of erythroid enhancer 5'HS2 is necessary for the high level expression of human beta E-globin gene in transgenic mice, indicating that the cis-element 5'HS2 is effective for abnormal beta-globin gene as well. Although the expression level of beta-globin gene was copy number-dependent as reported in previous studies, the average expression level per gene copy (12.1%) in transgenic mice bearing many copies of 5'HS2 beta E (16 copies) was significantly lower than that (79.7%) in transgenic mice bearing fewer copies of the gene (2 copies). The possible mechanisms for this decrease of expression are discussed. Novel hemoglobin tetramers which presumably consist of human beta E-globin chains and mouse endogenous alpha-globin chains have been assembled in transgenic mice.