Lead (Pb) produces aggresome-like inclusion bodies (IBs) in target cells as a toxic response. Our prior work shows metallothionein (MT) is required for this process. We used MT-I/II double knockout (MT-null) and parental wild-type (WT) cell lines to further explore the formation process of Pb-induced IBs. Unlike WT cells, MT-null cells did not form IBs after Pb exposure. Western blot of cytosol showed soluble MT protein in WT cells was lost during Pb exposure as IBs formed. Transfection of MT-I into MT-null cells allowed IBs formation after Pb exposure. Considering Pb-induced IBs may be like disease-related aggresomes, which often contain alpha-synuclein (Scna), we investigated Scna expression in cells capable (WT) and incapable (MT-null) of producing IBs after Pb exposure. Scna protein showed poor basal expression in MT-null cells. Pb exposure increased Scna expression only in WT cells. MT transfection increased Scna transcript to WT levels. In WT or MT-transfected MT-null cells, Pb-induced Scna expression rapidly increased and then decreased over 48 h as Pb-induced IBs were formed. A direct interaction between Scna and MT was confirmed ex vivo by antibody pulldown assay where the proteins coprecipitated with an antibody to MT. Pb exposure caused increased colocalization of MT and Scna proteins with time only in WT cells. In WT mice after chronic Pb exposure Scna was localized in renal cells containing forming IBs, whereas MT-null mice did not form IBs. Thus, Scna could be component of Pb-induced IBs and, with MT, may play a role in IBs formation.

The role of CpG methylation in the regulation of tissue-specific gene expression is highly controversial. Cyclin A1 is a tissue-specifically expressed gene that is strongly methylated in non-expressing tumor cell lines. We have established a novel real-time PCR method to quantitate genomic CpG methylation of the cyclin A1 promoter. Genomic DNA samples from different human organs were treated with bisulfite and amplified with methylation-specific primers and with primers amplifying methylated as well as non-methylated DNA. PCR product quantitation was obtained by using a fluorogenic probe labeled with FAM and TAMRA. These analyses demonstrated that the human cyclin A1 promoter was methylated in kidney, colon, spleen, testis, and small intestine, but not in brain, liver, pancreas, or heart. Expression of cyclin A1 was predominantly found in testis. Low level expression of cyclin A1 was present in spleen, prostate, leukocytes, colon, and thymus. Taken together, our data provide evidence that CpG methylation patterns of the human cyclin A1 promoter in human organs do not generally correlate with cyclin A1 gene expression in vivo. ß

Background Fall in elderly is a major public health problem. Characterizing trends in fall mortality in different subpopulations could help identifying the needs and developing preventive program for target groups. Here we evaluated the trends of fall-related deaths in Chinese mainland among adults aged ≥60 years specific in sex, age, and provinces, to measure the change in this mortality rate between 2013 and 2020, and to explore the underlying factors influencing this change.Methods Mortality data were retrieved from the National Disease Surveillance Points system(DSPs) of China, a national-level and provincial-level representative data source, to estimate the impact of elderly falls on mortality in the mainland of China and the specific provinces from 2013 to 2020. The joinpoint regression model was used to estimate the temporal trend of mortality in elderly fallen by calculating the annual percentage change (APC).Findings The age-standardized falls mortality was 10¢438 per 100 000 in 2020. The age-standardized mortality of elderly falls in total and female showed a steady increasing trend (APC=1¢96%, p = 0¢023 total; APC=3¢42%, p = 0¢003 female), with it was stable in males (APC=1.26%, p>0¢05). Fall mortality among the elderly was more common in people over 70 years of age and increased sharply. The death rates and APCs were highest among the oldest age groups(aged≥85 years). The higher fall mortality was mainly focused in the southeast and central regions, and lower rates were in the northeast provinces and Tibet.Interpretation Since 2013, the overall fall-related mortality trend among individuals aged ≥60 years has been consistently increasing in China, making it most critical public health challenge. Adherence interventions and increased social support for those at most risk should be considered.

Spider silks are attractive biopolymers due to their excellent mechanical properties and biomimetic potential. To optimize the electrostatic interaction for lysosomal drug delivery, a spider-eggcase-silk protein was genetically engineered using 5Â His Tag with a tailor-made isoelectric point of 4.8. By a facile HFIP-on-oil method, silk spheres were assembled as rapidly as 10 s. After the post-treatment of ethanol, silk spheres were determined with an improved compressive modulus by AFM indentation.Under incubation of silk spheres in a Doxorubicin solution, a maximum of 35% loading and average of 30% loading efficiency were determined. In the cytotoxicity experiment, silk spheres exhibited intrinsic biocompatibility and showed good control of the loaded drug in the neutral PBS solution. Significantly, by 96 h, the accumulative drug release at pH 4.5 was approximately 4.5-fold higher than that at pH 7.4. By conducting the platelet adhesion and hemolysis assay, Doxorubicin-loaded silk spheres exhibited good hemocompatibility. To further demonstrate this release behavior, within 24 h, Doxorubicin-loaded silk spheres were efficiently delivered to lysosomes and then released the payload to the nuclei of Hela cells. rsc.li/rsc-advances 9394 | RSC Adv., 2018, 8, 9394-9401 This journal is