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Objective To investigate the effect of DNA damage protein targeting VCP (DVC1) in DNA protein crosslink repair induced by nitrogen mustard in human bronchial epithelial cells (16HBE). Methods DVC1 siRNA was used to interfere its expression in 16HBE. Then the cells were exposed to 50 μmol/L nitrogen mustard, and the expression levels of DNA damage repair proteins were detected by Western blotting. The total content of DNA-protein crosslink was determined by fluorescence microplate analyzer from 1 to 24 h. The crosslink content of DNA with O6-methylguanine-DNA methyltransferase (MGMT), a methyltransferase, was detected by Slot blotting. The level of ubiquitinated MGMT was detected by immunoprecipitation. After GST-DVC1 fusion protein was constructed, GST pull-down was used to detect the binding of DVC1 with MGMT and ubquitin. Results After si-DVC1 intervention, the total content of DNA-protein crosslink was increased, and exposure to nitrogen mustard decreased the level of prototype of MGMT, but increased the level of ubiquitinated MGMT. DVC1 could bind to ubquitin, and si-DVC1 intervention further enhanced the ubiquitination of MGMT, as well as the crosslink between MGMT and DNA, which was similar with NSC697923 (an E2 ubiquitin-conjugating inhibitor) and MG132 (a proteasome inhibitor) treatment. The expression of Rad51 and Ku70, 2 DNA repair proteins, were increased and decreased respectively by si-DVC1 interference. Conclusion Clearance of DNA-protein crossslink is depend on ubiquitination of MGMT. DVC1 promotes the clearance of nitrogen mustard induced DNA-protein crossslink by interacting with ubiquitin, and maintain the ability of homologous recombination repair.

Chemical, biological, radiological, and nuclear (CBRN) defense training is an essential means to deal with CBRN threats, and is given great importance by military forces in different countries. This article summarizes CBRN training methods adopted by foreign armies, and introduces various CBRN training methods including computer simulation training, base training, simulation equipment training, simulated chemical warfare chemical agent training, station-touring training, non-military scenario training, and transnational training. In the CBRN training, foreign armies emphasize internet- and computer-based training, the establishment of high level training bases and scenario-simulating equipment, the application of station-touring training, and the combination with non-military conditions. The practices of foreign military forces provide valuable lessons for improving the techniques, approaches and the overall level of CBRN training quality in China.

Objective To investigate the effect of vitamin D3 on nitrogen mustard (NM)-induced keratinocyte inflammation and the role of NOD-like receptor protein 3 (NLRP3) inflammasome in the process. Methods HaCaT cells were treated with 20 μmol/L NM, in the presence or absence of 10 μmol/L MCC950 (a specific inhibitor of NLRP3 inflammasome) or 10 μmol/L Ac-YVAD-cmk (YVAD, a specific activator of caspase-1) or 5 nmol/L vitamin D3 for 4 h. CCK-8 assay was used to detect cell viability. The IL-1β content in the supernatant of cell lysate was determined by Human IL-1β ELISA kit, and the expression of NLRP3, caspase-1, IL-1β and COX-2 was detected by Western blotting. Results Treatment with 20 μmol/L NM had no significant effect on the viability of HaCaT cells, but remarkably up-regulated the expression of NLRP3, caspase-1 p20, IL-1β and COX-2, and enhanced the accumulation of IL-1β in the culture supernatant of HaCaT cells (P < 0.05). Moreover, MCC950 or YVAD treatment abolished the above effects of NM (P < 0.05). Simultaneously, vitamin D3 treatment notably inhibited the upregulated expression of NLRP3 and caspase-1 p20, and remarkably reduced the increased expression and release of IL-1β and COX-2 in HaCaT cells induced by NM (P < 0.05). Conclusion Vitamin D3 ameliorates NM-induced keratinocytes inflammation by reducing the synthesis of inflammatory mediators through suppressing the activation of NLRP3 inflammasome

Aging‐associated renal dysfunction promotes the pathogenesis of chronic kidney disease. Mitochondrial dysfunction in renal tubular epithelial cells is a hallmark of senescence and leads to accelerated progression of renal disorders. Dysregulated calcium profiles in mitochondria contribute to aging‐associated disorders, but the detailed mechanism of this process is not clear. In this study, modulation of the sirtuin 1/angiotensin II type 1 receptor (Sirt1/AT1R) pathway partially attenuated renal glomerular sclerosis, tubular atrophy, and interstitial fibrosis in D‐galactose (D‐gal)‐induced accelerated aging mice. Moreover, modulation of the Sirt1/AT1R pathway improved mitochondrial dysfunction induced by D‐gal treatment. Transient receptor potential channel, subtype C, member 3 (TRPC3) upregulation mediated dysregulated cellular and mitochondrial calcium homeostasis during aging. Furthermore, knockdown or knockout (KO) of Trpc3 in mice ameliorated D‐gal‐induced mitochondrial reactive oxygen species production, membrane potential deterioration, and energy metabolism disorder. Mechanistically, activation of the AT1R/PKA pathway promoted CREB phosphorylation and nucleation of CRE2 binding to the Trpc3 promoter (−1659 to −1648 bp) to enhance transcription. Trpc3 KO significantly improved the renal disorder and cell senescence in D‐gal‐induced mice. Taken together, these results indicate that TRPC3 upregulation mediates age‐related renal disorder and is associated with mitochondrial calcium overload and dysfunction. TRPC3 is a promising therapeutic target for aging‐associated renal disorders. [ABSTRACT FROM AUTHOR]

Using sulfur mustard analog 2-chloroethyl ethyl sulfide (CEES), we established an in vitro model by poisoning cultured immortalized human bronchial epithelial cells. Nile Red staining revealed lipids accumulated 24 h after a toxic dose of CEES (0.9 mM). Lipidomics analysis showed most of the increased lipids were triglycerides (TGs), and the increase in TGs was further confirmed using a Triglyceride-Glo™ Assay kit. Protein and mRNA levels of DGAT1, an important TG biogenesis enzyme, were increased following 0.4 mM CEES exposure. Under higher dose CEES (0.9 mM) exposure, protein and mRNA levels of PPARγ coactivator-1ɑ (PGC-1ɑ), a well-known transcription factor that regulates fatty acid oxidation, were decreased. Finally, application with DGAT1 inhibitor A 922500 or PGC1ɑ agonist ZLN005 was able to block the CEES-induced TGs increase. Overall, our dissection of CEES-induced TGs accumulation provides new insight into energy metabolism dysfunction upon vesicant exposure. In CEES (0.9 mM)-injured cells: Triglycerides (TGs) were abundant in the accumulated lipids. Expression of DGAT1, not DGAT2, was increased. Expression of PGC1ɑ, not PGC1β, was reduced. DGAT1 inhibitor or PGC1ɑ agonist blocked the CEES-mediated increase in TGs. [ABSTRACT FROM AUTHOR]