目的 研究SD大鼠反复暴露于84消毒液对多种器官的伤害情况.方法 SD大鼠30只,雌雄各半,在密闭空间内分别按100 mL/m2喷洒含0(对照组)、400、10000 mg/L次氯酸钠的84消毒液,大鼠暴露2次/d,每次1 h,持续28 d,腹主动脉穿刺收集血样,用于血液学和血液生化指标检测,于第29天,所有大鼠麻醉后实施安乐死,采集肝、肾、心、脾、肺、气管、眼球和皮肤等重要脏器进行进一步检查.结果 高剂量组大鼠重复染毒28 d后,气管、肺、肝、肾、血液系统出现病变,眼和呼吸道出现急性刺激症状,血细胞增多和凝血功能障碍,气管、肺、肝和肾发生病理学改变.低剂量组和对照组无异常改变.结论 长期大剂量接触84消毒液可能对动物机体健康造成不良影响,在使用中应加强个人防护.

线粒体是细胞能量产生、维持细胞代谢和内稳态以及调节信号通路的重要结构.药物可引起线粒体氧化应激,干扰线粒体毒性通路,损伤线粒体功能.线粒体功能受多条毒性通路的调控,抗氧化应激通路HO-1/PGC-1α在线粒体氧化应激中发挥重要作用.该文综述了氧化应激对毒性通路的影响以及HO-1/PGC-1α通路在调控线粒体氧化应激中的作用.

目的:探讨氟他胺对人肝细胞线粒体生物合成的影响以及抗氧化通路Nrf2的调节作用.方法:采用人源肝细胞HepG2,氟他胺(0～50μmol/L)给药24 h,通过RT-PCR和Western blot方法,检测mtDNA拷贝数和线粒体生物合成关键蛋白的表达,再通过应用基因敲降和特异性激活剂或抑制剂等技术方法进一步观察ERK1/2对氟他胺诱导的线粒体生物合成的影响以及Nrf2通路的调控作用.结果:氟他胺可诱导线粒体生物合成,mtDNA拷贝数和ERK1/2、PGC-1α蛋白均呈现剂量依赖性上升;ERK1/2抑制和激活能改变氟他胺诱导的mtDNA拷贝数和PGC-1α 的表达;Nrf2通路抑制可影响氟他胺诱导的mtDNA拷贝数和ERK1/2、PGC-1α 的表达.结论:氟他胺可影响线粒体生物合成,其机制与Nrf2介导的ERK1/2改变密切相关.

药物性肝损伤(DILI)是临床上一类常见的肝病变,已成为药物研发失败或药品撤市的重要原因.有害结局路径(AOP)作为将化学物质致毒的分子起始事件和一系列关键毒性反应到最终有害结局串联起来的框架,为化学物毒性评价和风险评估提供了新工具.近年来,AOP在DILI等肝损伤评估中得到越来越多的发展和应用,目前AOP知识库已收录25条肝损伤相关AOP.本文介绍AOP的内涵和基本框架,重点综述了肝损伤相关AOP的分子起始事件,代谢酶功能异常、线粒体损伤、炎症反应和免疫反应等关键事件,以及肝纤维化、脂肪变性、胆汁淤积和肝癌等有害结局,并从AOP网络和定量AOP介绍未来AOP的发展趋势,为建立并完善更多的肝损伤相关AOP提供数据资料和研究思路,以期推动AOP在DILI等肝损伤风险评估中的应用.

新型冠状病毒肺炎(简称“新冠肺炎”)疫情是一场严重危害人类健康以及经济和社会发展的重大疫情.化学消毒剂是有效阻击新冠肺炎疫情蔓延、实现疫情精准防控的重要手段.然而,过量使用化学消毒剂以及防护意识薄弱已成为当前疫情防控中较为关注的问题.这些化学消毒剂本身均具有一定的毒性,特别是过量使用时有可能导致明显的健康危害甚至危及生命安全,其毒性风险值得高度重视和警惕.

目的 了解新型冠状病毒肺炎期间武汉地区医院消毒剂的使用情况.方法 采用自行设计问卷和现场调研等方法调查了疫情较为严重的武汉地区医院的消毒剂使用情况,调查内容包括消毒剂的种类、使用频次、使用量及使用范围等.结果 新冠肺炎期间最常用的消毒剂是含氯类和醇类消毒剂;每天工作场所中消毒频率以3次(52.85％)及3次以上(22.36％)居多,个人每天手部使用消毒剂在10次以上的达到182人(73.98％).大部分人(210人,85.37％)按照说明书的浓度配制使用消毒剂,超过一半的人(137人,55.69％)所在工作场所中单次消毒剂使用量>500 mL.截止至3月,工作场所累计消毒时间几乎全部超过10 d以上.216人(87.80％)所处的工作环境为全方位消毒(空气、物体表面和地面消毒).结论 建议在消毒过程中按照合理使用和按需配制的原则,最大限度保障医护人员和就医人群的身体健康.

目的 调查分析新冠肺炎疫情期间部队营区消毒剂的使用及防护情况,发现问题并提出建议对策.方法 主要采用问卷调查方式,基于互联网平台对北京、武汉、西安等地的部队营区进行消毒剂使用和防护调查,并进行统计分析.结果 通过问卷了解到新冠疫情期间营区进行消杀作业最常用的消毒剂是含氯类(84等)消毒剂和75％乙醇,大多数工作场所消毒每天集中消毒2次(48.39％)或3次(36.69％);单次消毒剂使用量主要集中在10-100 ml(41.13％),能够按照说明书浓度配制消毒剂的占比90.32％;消毒范围以全方位消毒为主(46.37％),包括空气消毒、物体表面消毒和地面消毒;截止到4月中旬,大部分营区累计消毒时间已超过60天(70.97％);营区内专业消杀作业人员培训率为57.72％,经过培训的人员防护率明显高于未培训人员(P<0.05).结论 部队营区消毒剂使用存在过度消毒及培训不足、防护不规范等现象,部分单位有待加强消毒作业规范和防护指导.

目的 研究镉对LNCaP细胞氧化应激的影响以及活性氧(ROS)在镉诱导SENP1表达中的作用.方法 取对数生长期LNCaP细胞,分别以终浓度为0、1、4和8μmol/L的氯化镉处理24 h后,采用荧光显微镜和流式细胞仪测定细胞内ROS水平,采用比色法检测细胞内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,采用Western blot检测细胞内雄激素受体(AR)、去类泛素化酶1(SENP1)、以及氧化应激相关信号通路分子和转录因子IKKα/β、HO1、AKT、ERK1/2、P38、Nrf2的表达水平,进一步对细胞核内AR、Nrf2的蛋白表达水平进行检测;用ROS清除剂NAC预处理LNCaP细胞后,再进行氯化镉处理24 h,然后采用Western blot检测细胞内SENP1表达水平.结果 LNCaP细胞在镉处理后,与对照组相比细胞内ROS水平明显升高,差异有统计学意义(P＜0.001),镉处理组SOD活性降低,差异有统计学意义(P＜0.01),GSH-Px活性升高,差异有统计学意义(P＜0.05);AR、SENP1、p-IKKα/β、HO1、p-AKT、p-ERK1/2和p-P38的表达水平随处理组镉剂量的增加而升高,核内AR、Nrf2蛋白表达水平也随之升高;镉可以增强SENP1的表达,而ROS清除剂预处理后,这种增强作用减弱.结论 镉可能通过增加细胞内ROS水平,进一步激活AKT、ERK、P38和NF-κB信号通路,增强SENP1的表达,促进AR和Nrf2入核,从而导致前列腺癌的发生.

间充质干细胞(MSC)是可在体外贴壁培养的、具有多向分化潜能的成体干细胞,可从多种组织中分离得到.研究表明,MSC具有组织修复、免疫调节、支持造血及旁分泌等生物学特性,目前已有大量研究将其应用于对中枢神经系统、循环及造血系统疾病、移植物抗宿主病等疾病的治疗.该文综述了MSC在军事医学领域治疗急性放射病、造血系统损伤、胃肠综合征、皮肤损伤及混合伤等伤病的研究进展及应用前景.

Objective To investigate the toxic effects and mechanisms of sulphur mustard on DNA damage of human immortalized epidermal keratinocytes (HacaT cells).Methods The inhibitory effect of sulphur mustard on the proliferation of HacaT cells was detected by CCK-8 method.The apoptosis index of cells was measured by Annexin V-FITC method.The effects of sulphur mustard on DNA damage were detected by single cell gel electrophoresis.The expression levels of DNA damage and repair related proteins were detected by Western blot.Results The proliferation rate of HacaT cells was inhibited in a dose-dependent manner after treatment with sulphur mustard for 24 h (IC50 value was 121 mol/L).The apoptotic and comet tailing rates of cells treated with sulphur mustard also increased in a dose-dependent manner.The expression levels of DNA damage and repair related proteins were changed after treatment with sulphur mustard.Conclusion Sulphur mustard has significant cytotoxic effect on HacaT cells,and can induce apoptosis and DNA damage.In addition,ATM-P53-γH2AX-PARP signaling pathway plays an important role in the repair of DNA damage induced by sulphur mustard.

OBJECTIVE:To study the oxidative damage of PM2.5 in human embryonic stem cells (EBf-H9 cells), and to provide a theoretical basis for revealing the adverse health effects of PM2.5 and the potential mechanisms, and also to provide a new alternative cell model for PM2.5 risk assessment.METHODS:EBf-H9 cells were cultured with 0.00 (the constrast group) 3.91, 7.81, 15.63, 31.25, 62.50, 125.00 μg/cm(2) of PM2.5. CCK-8 kit was used to determine the survival rate of cells exposed to PM2.5 for 6 h. DCFH-DA probe was used to detect the total ROS content of cells exposed to PM2.5 for 1 h. The activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), and the content of lipid peroxides such as malondialdehyde (MDA) in cells exposed to PM2.5 for 6 h were detected by using the commercial kits. ANOVA model analyzed the statistical significance from the different concentration group.RESULTS:The cytotoxicity results showed that the cell survival rate was decreased gradually with the increase of the concentrations of PM2.5, and the half inhibitory concentration (IC50) was 83.01 μg/cm(2). When the exposure concentration was 3.91, 7.81, 15.63, 31.25, 62.5 μg/cm(2), after exposure of PM2.5 1 h, the ROS florescence was 27.12±0.21, 54.03±0.50, 60.93±0.08, 61.36±1.00, 68.21±0.93, 78.27±1.26 (compared to control group 27.12±0.21, all P level<0.01). After exposure of PM2.5 6 h, the activities of T-SOD was (9.78±0.28), (8.59±0.22), (8.90±0.33), (7.46±0.71), (4.21±0.17) U/mg protein (F=98.881, compared to control group (11.77±0.63) U/mg protein, all P level<0.01). The activities of GSH-Px was (181.59±3.65), (153.33±1.69), (168.74±2.22), (81.56±0.56), (48.62±2.13) U/mg protein (compared to control group (273.90±6.50) mU/mg protein, all P level<0.01). And the content of MDA was (0.38±0.03), (0.43±0.09), (0.47±0.09), (0.65±0.10), (0.70±0.12) nmol/mg protein (compared to control group (0.27±0.02) nmol/mg protein, all P level<0.05).CONCLUSION:PM2.5 exposure can decrease EBf-H9 cells viability, and improve the levels of lipid peroxidation. It may be due to induce EBf-H9 cells to increase the production of ROS and to make the cells appear oxidative stress, which lead to oxidative damage to cells. The present study reveals the mode of action of PM2.5 in terms of oxidative damage to EBf-H9 cells. It is also indicated that the cells may be a new alternative cell model for PM2.5 risk assessment.

Objective To observe toxic symptoms and signs , toxic damage extents and reversibility in rats after oral administration of Tangwang Mingmu granules .Methods Four dose groups with 40 rats in each group were designed in this study, including control group fed with distilled water and three groups at different dosages of the test drug .Tangwang Mingmu granules were orally administered to SD rats at the dosage of 8.4, 4.2 and 2.1 g/kg for 3 weeks and 14.0, 8.4 and 4.2 g/kg for 23 weeks, for 26 consecutive weeks .The general state of the rats was observed every day , while body mass and food consumption were calculated once a week .Halfway through and at the end of the administration (13 and 26 weeks) and after four weeks of recovery, parameters of body mass, hematology, hematological biochemistry, organ/body mass ratio and histopathology were measured .Results Compared with the control group at the same time-point, body mass of male rats in the other three groups was slightly reduced .Food consumption in high and medium dose groups was reduced (P<0.05), MCHC, ALT, TBIL and Na +in high dose group were decreased (P<0.05), TP, ALB and D-BIL were increased (P<0.05), the mean body mass and relative organ weight of thymus in medium dose male rats were decreased (P<0.05), relative organ weight of the liver and kidney in high dose male rats was increased (P<0.05), and focal chronic inflammation to different extent was observed in the liver , kidney and prostate gland .No dose-effect relationship was found in these perturbations that were all within the normal range of animals .No significant drug-related pathological changes were found.Conclusion The NOAEL of Tangwang Mingmu granules is considered to be 14.0 g/kg body mass/day (equal to 50 times the proposed clinical adult dosage ) for the 26-week repeated dose oral toxicity study in male andfemale rats.

目的 分析不同种类的消毒产品一般毒理学安全性评价结果,为消毒产品的安全性评价的剂量设计和毒性分级提供参考.方法 采用统计分析方法,对本研究室急性毒性和亚急性毒性试验的41个化学消毒剂产品试验结果进行分析和评价.结果 急性经口毒性试验评价的41个消毒产品中,属实际无毒级的消毒产品占总数的73.2％,属低毒级的消毒产品占总数的26.8％.亚急性毒性试验评价的41个消毒产品中,无可见有害作用水平(NOAEL)为1 000 mg/kg的消毒产品占总数的66.7％,NOAEL为500～1 000 mg/kg的消毒产品占总数的13.3％,NOAEL为100～500 mg/kg的消毒产品占总数的6.7％,NOAEL为10～ 100 mg/kg的消毒产品占总数的13.3％.结论 醇类、碘类、胍类消毒产品属实际无毒级,较为安全;过氧化物类、含氯类、醛类消毒产品毒性相对较大.

人诱导多能干细胞与人胚胎干细胞相似,在体外可分化为各种类型的体细胞,其来源充足,可针对个人或某种疾病取材,为药物早期毒性评价体外替代方法提供了一个新的可选细胞模型.目前,利用人诱导多能干细胞获得的心肌细胞用于药物引起的心率改变、QT间期延长和心肌损伤等心脏毒性评价;利用其获得的神经细胞,结合高通量、高内涵技术及电生理学技术,可用于药物引起的神经突出生长异常、电生理改变及神经发育毒性评价;利用人诱导多能干细胞可获得个体特异性的大量肝细胞,具有较高的细胞色素P450酶活性,能够比较真实的反映人肝细胞的代谢特征,用于评价药物肝细胞毒性;人诱导多能干细胞具有多能性,在体外可分化为外、中和内胚层,具有用于发育毒性评价的可能性,对三胚层相应标志分子的检测有助于发育毒性评价终点的确立;人诱导多能干细胞还可用作3D培养的种子细胞,构建三维立体组织和器官模型,用于药物早期毒性评价,进一步缩小细胞水平与体内水平评价结果的差异.

目的 建立消毒产品亚急性毒性试验SD大鼠血液生化正常参考值范围.方法 采用全自动生化分析仪及血细胞分析仪检测方法,对模型SD大鼠的12项血清生化指标及18项血液学指标进行评价,建立正常参考值范围.结果 SD大鼠10项血清生化指标雌雄性别差异显著(P＜0.05),其中丙氨酸氨基转移酶等3项指标为雄性大鼠高于雌性;总蛋白等7项指标为雄性大鼠低于雌性.SD大鼠8项血液学指标也存在雌雄性别差异(P＜0.05),除平均红细胞血红蛋白浓度雄性大鼠低于雌性之外,白细胞等其余7项血液学指标均为雄性高于雌性.结论 试验模型大鼠中多数血液生化指标存在雌雄性别差异,本研究建立的SPF级SD大鼠的血液生化正常参考值,可为消毒产品毒性评价提供参考.

通过研究空气颗粒物的代表性组分纳米碳黑(nano particle carbon black,NPCB)与重金属(Pb/Cr/Cd)联合染毒对BEAS-2B细胞存活率和LDH漏出率的影响,旨在阐明NPCB与重金属对细胞毒性的联合作用模式.检测NPCB与重金属(Pb/Cr/Cd)联合染毒24 h后BEAS-2B细胞存活率(CCK-8法)和LDH漏出率(LDH活性比色法)的变化,采用析因方差分析判断其是否存在联合毒性作用及联合作用模式.NPCB与重金属(Pb/Cr/Cd)联合染毒在细胞存活率和LDH漏出方面存在联合作用;与对照组和单独染毒组相比,低剂量Pb(125μmol·L-1)与NPCB联合染毒对细胞存活率无交互作用,对LDH漏出表现为拮抗作用;高剂量Pb(1000μmol·L-1)与NPCB联合染毒对细胞存活率表现为协同作用,对LDH漏出无交互作用;Cr和Cd与NPCB联合染毒在细胞存活率方面均表现为协同作用;低剂量Cr和Cd与NPCB联合染毒在LDH漏出方面无交互作用,高剂量时表现为协同作用.NPCB与重金属存在联合作用,金属不同、剂量不同以及评价指标不同,其联合作用模式不尽相同.

OBJECTIVE ToexplorethepossiblemechanismoractiontargetsofT-2toxinembryo toxicity by observing the effect of T-2 toxin on mitochondrial function of differentiated murine e mbryonic stemcells(mESCs).METHODS Duringdifferentiationat24,72and120h,ESCswereexposedto T-2 toxin 0.5 μg·L-1 .Meanwhile,mESCs were pre-treated with antioxidant Trolox (200 μmol·L-1 )for 30 min and exposed to T-2 toxin (0.5 μg·L-1 )for 72 h.The mitochondrial ultrasture of differentiated mESCs was observed under a transi mission electrical microscope (TEM).The differentiated ESC mito-chondrial function,including respiratory control ratio (RCR),ATP synthase activity and mitochondrial membranepotential(MMP),wasmeasuredat144hafterdifferentiation.RESULTS Significant decrease of the mitochondrial number,deformation of mitochondrial structure,and lack of complete mito-chodrial crest were observed through TEM in the groups of T-2 toxin exposed for 72 and 1 20 h,respec-tively.Compared with the normal control group,RCR declined by 49.5% and 55.1%,ATP synthase activity decreased by 84.9% and 89.3%,and MMP decreased by 23.2% and 35.2% in T-2 toxin 0.5 μg·L-1 exposure 72 and 1 20 h group,respectively.However,the inhibition of mitochondrial function by T-2 toxin in differentiated mESCs recovered significantly in the presence of the antioxidant Trolox. CONCLUSION T-2toxininducesoxidativestressandinhibitsmESCsmitochondrialfunctionindifferenti-ated mESCs,and ROS-induced mitochondrial malfunction plays an i mportant role in T-2 toxin e mbryonic toxicity mechanis m.

目的 评价季铵盐类消毒剂毒性,为实际应用提供安全性依据.方法 采用动物试验方法,对一种固体季铵盐消毒剂进行了一般毒性实验研究.结果 该季铵盐消毒剂对小鼠急性经口LD50为762.2 mg/kg,大鼠急性经口LD50为986.0 mg/kg.亚急性毒性试验各剂量组动物体重、血细胞计数及分类、血生化指标与对照组比较均无显著性差异,动物主要脏器未见明显病理变化.结论 该季铵盐消毒剂的急性作用主要与其消化道直接毒性有关,对大小鼠属低毒;未发现该季铵盐消毒剂有明显亚急性毒性作用.

目的 应用小鼠胚胎干细胞(mESC)作为模型探讨T-2毒素对mESC的作用靶点与作用机制.方法 选用0.5 ng/ml T-2毒素处理分化过程中的mESC 24、72和120 h,流式细胞术检测mESC的活性氧簇(ROS)的生成,比色法检测mESC的抗氧化防御酶活性,包括超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPx)的活力变化,以及脂质过氧化产物丙二醛(MDA)的含量.结果 0.5 ng/ml T-2毒素作用72与120 h,mESC中ROS大量蓄积,抗氧化防御酶(SOD、GPZ)活力下降,MDA增加,其效应呈现时间依赖关系.自由基清除剂(Trolox,200 μmol/L)预处理可以降低T-2毒素对mESC抗氧化防御酶活力的抑制作用,减轻T-2毒素引起的mESC中ROS的蓄积以及MDA的增加.结论 低剂量T-2毒素染毒引起mESC的氧化损伤是其发育毒性的重要机制之一.

OBJECTIVE To observe the modulation of resveratrol on cardiomyocyte differentiation of murine embryonic stem cells(ESCs) and investigate the underlying mechanism.METHODS The murine ESCs were differentiated as embryonic bodies(EB) in hanging drops and treated by resveratrol 0,0.44,4.4 and 44 μmol·L-1 for 96 h.The number of EB containing spontaneously contracting cells was recorded under the light microscope.The mitochondrial ultrastructure was observed in transmission EM photos.The gene expression of α-myosin heavy chain(α-MHC),peroxisome proliferator activated receptorγ(PPARγ),PPARγ coactivator 1α(PGC-1α),nuclear respiratory factor-1(NRF-1),mitochondrial transcription factorA(mtTFA) and mitochondrial respiratory chain complexⅣ(COXⅣ) was detected by real-time PCR while the expression of α-actinin,PPARγ and PGC-1α proteins was measured by Western blotting.RESULTS Resveratrol 0.44 and 4.4 μmol·L-1 increased the number of contracting myocardial cells in differentiated ESCs.Compared with normal control group,the expression of cardiac-specific α-MHC genes at the concentration of 0.44 and 4.4 μmol·L-1 resveratrol increased 6,6-fold and 4.7-fold,and cardiac-specific α-actinin protein increased 2.7-fold and 3.1-fold.Furthermore,the increase in the number of mitochondria was observed in transmission EM photos of differentiated ESCs treated by resveratrol 4.4 μmol·L-1.PPARγ gene and protein expression were increased in all groups treated by resveratrol at different concentrations.Meanwhile,the expression of genes correlated with mitochondrial biogenesis increased significantly after treatment by resveratrol,especially at the concentration of 0.44 and 4.4 μmol·L-1.CONCLUSION Resveratrol induces murine ESC differentiation to cardiomyocytes in vitro by agitating PPARγ and promoting mitochondrial biogenesis modulated by PGC-1α.