Research progress in antidotes of nerve agents in the USA / 军事医学

Nerve agent not only inhibit acetylcholinesterase ( AChE) at an early stage, but also induce prolonged and progressive neuroinflammation and delayed neurodegeneration.Recently, the US National Institute of Health ( NIH) has sponsored some major programs of toxic mechanisms and treatment of nerve agents, which aims at the development of quick and effective treatment to acute intoxication and delayed effect.The experimentally effective new antidotes mainly include AChE-targeting drugs, broad-spectrum reactivators and scavengers, antiinflamatory and nerve protection drugs.

Effect of glucose concentrations on energy metabolism in 16 HBE cells exposed to CEES / 军事医学

Objective To compare the changes in energy metabolism in 2-chloroethyl ethryl sulfide(CEES)-poisoned bronchial epithelial cell 16HBE cultured in media at different glucose concentrations .Methods Bronchial epithelial cell 16HBE was cultured in high (4.5 mg/ml) or low (1.1 mg/ml) glucose medium and exposed to a sulfur mustard simulant CEES of 0.2, 0.5, 1.0 mmol/L.Cell growth and cytotoxicity were tested using MTS .ATP, ADP and AMP were detected by HPLC and the value of ATP/ADP, total adenine nucleotides ( TAN) and energy charge ( EC) was subsequently calculat-ed.Mitochondrial oxidative phosphorylation-related proteins, COX-10 and ISCU, were detected using Western blotting . Rhodamine 123 was applied to detect the mitochondrial membrane potential using flow cytometry .Results Low glucose accelerated the growth and energy metabolism of 16HBE cells in regular culture , and the contens of ADP , TAN, COX-10 and ISCU in low glucose group were significantly higher than those in high glucose group .CEES exposure (≥0.5 mmol/L) significantly affected cell viability in both high and low glucose groups , with significant difference between the two groups exposed to 1.0 mmol/L CEES.In high glucose group, 24 h after 0.5 or 1.0 mmol/L CEES exposure, the contents of ATP, ADP and TAN were significantly increased , while ATP/ADP and EC decreased .In low glucose group , ADP, AMP and TAN significantly decreased, while ATP/ADP and EC increased 24 h after 1.0 mmol/L CEES exposure.The mi-tochondrial membrane potential (MMP) also changed differently after 0.5 mmol/L CEES exposure.MMP in high glucose group marginally increased at 3 h, and significantly increased at 8-12 h (P<0.05), and returned to normal at 24 h. MMP in low glucose group showed a transient decrease at 5 h (P<0.01), and back to normal at 8 h.The protein levels of COX-10 and ISCU were significantly increased in high glucose group 24 h after 0.5-1.0 mmol/L CEES exposure , but sig-nificantly decreased in low one 24 h after 1.0 mmol/L CEES exposure .Conclusion When 16HBE is cultured at a high or low glucose concentration , the cell growth, stress responses and energy metabolism including MMP , COX-10, ISCU and ATP production are in different status before or after CEES exposure .High glucose could protect against CEES exposure .

miR-34a partially reverses inhibition of CEES-exposed keratinocytes migration via ERK1/2 pathway / 军事医学

Objective To explore the effect of 2-chloroethyl ethyl sulfide(CEES) poisoning on keratinocyte migration and the regulatory role of microRNA(miR)-34a.Methods MTS was used to detect the viability of cells exposed to CEES in order to select an appropriate dose of CEES exposure in this in vitro model.The protein level of keratin 5 and keratin 10 was detected to assess cell differentiation status .Scratch assay was applied to evaluate cell migration ,and miR-34a silencing in keratinocytes was achieved by transfecting chemically synthesized miR-34a specific miRNA inhibitor.t-ERK1/2 and p-ERK1/2 levels closely related to cell migration were detected using Western blotting .Results An in vitro CEES exposure model of keratinocytes was established at the optimal concentration of 0.5 mmol/L CEES in the viability test , and this dose was chosen to evaluate cell migration changes .The migration of cells was significantly inhibited 24 h after CEES exposure , accompanied by no changes in morphology and keratin 5/10 levels.Silencing of miR-34a significantly increased the migration of cells exposed to CEES , which could be blocked by adding 5 μmol/L U0126 , an ERK1/2 phosphorylation selective inhibitor.Conclusion Silencing of miR-34a can significantly increase keratinocyte migration and partially reverse the inhibition of CEES-caused migration , which could be mediated by ERK 1/2 pathway activation .

Effects of hypoxia and sodium cyanide on superoxide dismutase,reduced glutathione and malondialdehyde in arterial blood of rabbits / 第三军医大学学报

Objective To investigate the effects of hypoxia and sodium cyanide(NaCN)on oxidative stress in rabbit arterial blood.Methods An artificial hypobaric hypoxia chamber was used to simulate 4 000-meter high altitude.Twenty rabbits were randomly divided into 4 groups:hypoxia group with high-or low-dose,non-hypoxia with high-or low-dose.The animals in the non-hypoxia groups were operated under normal circumstances while those in hypoxia groups were subjected to chamber in low pressure for 72 h before receiving the hypoxia experiments.Femoral arterial cannulation was performed on all animals under anesthetization with pentobarbital sodium(30 mg/kg,iv)and NaCN(ip)at the doses of 1.5 mg/kg and 2 mg/kg.Blood samples were collected at 10 min before intoxication,and 5,10,15,20,30,60,120 and 180 min after intoxication and blood seperation was conducted.The activity of superoxide dismutase(SOD),contents of reduced glutathione(GSH)and malondialdehyde(MDA)were determined.Results The MDA content was significantly increased(P

Effects of plateau hypoxia on the cyanide ion metabolism and correlative toxicant mechanism / 解放军医学杂志

Objective The aim of present study was to explore the effects of plateau hypoxia on the cyanide ion metabolism and correlative toxicant mechanism.Methods 12 adult rabbits and 72 male SD rats were randomly divided into two groups separately:plain control group with NaCN intoxication and plateau NaCN intoxicant group.2mg/kg NaCN was subcutaneously injected into the back of rabbits,the femoral vein blood was then collected at different designated time points for the measurement of cyanide,hemoglobin and ferrihemoglobin concentrations.Meanwhile 3.6 mg/kg NaCN was subcutaneously injected into the back of rats,the cardiac blood and the hepatic tissue were then collected at the different designated time points for the determination of cyanide ion,cytochrome oxidase activities and for the detection of pathologic changes of hepatic tissue.Results Under the condition of plain and plateau environments,the pharmacokinetics of rabbits induced by NaCN injection was characterized by one-compartment model.The blood cyanide concentration of rats in both plain and plateau groups reached peak value at 30min,and the activity of cytochrome oxidase decreased.Furthermore,the pathologic diagnosis of rats hepatic tissue suggested that liver injury induced by NaCN intoxication at high altitude was more serious than in plain intoxicant.Conclusion Hypoxia could markedly disturb the metabolic process of NaCN in vivo,aggravate the inhibition of cytochrome oxidase activity and lead to serious pathologic injury.