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检索条件"任意字段=2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会"

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Modeling the three-dimensional structure of embryonic brain based on its continuous sections

Modeling the three-dimensional structure of embryonic brain ...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Binbao Wu Suai Xu Zihao Shen Yuqiong Liu Xiaoping Chen College of Biotechnology and Bioengineering Zhejiang University of Technology

Purpose:Embryonic brain is a sensitive and important target for toxicity ***,the complex developmental change makes it difficult to be represented by a few separated tissue *** this study,we developed a computer software based method to transform hundreds of embryonic brain slices data into three-dimensional reconstruction *** study will provide foundation for our further alternative testing study in *** E16 and E18 mice brains were fixed in PFA,dehydrated in gradient alcohol and embedded in paraffin,then cut into continuous slices in coronal orientation.190 pieces of continuous slices from hippocampus region(E18 embryo) and 120 pieces of slices from striatum region(E16 embryo)were selected based on the anatomical atlas,and the slices were stained then with cresyl violet(Nissl staining).Images were acquired using an Olympus LX71 microscope with 2×10 ***,grayscale-adjustment and automatic registration of images were done using a Photoshop CS6 and Image Pro Plus software,followed with a manual *** adjusted images were imported into 3 D-doctor software to construct three-dimensional images,and the prepared models were saved as STL *** final stereoscopic models were represented using Cura software to show,the three-dimensional ***:In the reconstructed hippocampus model,the images were represented in one-tenth ratio of the primary *** voxel width of X axis was 70.2 mm,the voxel width of Y axis was 56.9 mm,and the voxel width of Z axis was 37.7 *** the reconstructed striatum model,the images were represented in one in fifty ratio of the primary *** voxel width of X axis was 36.4 mm,the voxel width of Y axis was 19.0 mm,and the voxel width of Z axis was 16.7 mm,***:The three-dimensional reconstruction of continuous sections of the embryonic brain regions represent the stereoscopic images of embryonic brain ***,it helps to s

关键词： embryonic brain continuous sections three-dimensional reconstruction Hippocampus Striatum

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Alternative Methods for Skin Sensitisation Risk Assessment

Alternative Methods for Skin Sensitisation Risk Assessment

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Yuan Gao Petra Kern Cindy.A.Ryan Frank Gerberick P&G Technology(Beijing) Co. Ltd. Procter & Gamble Services Company NV. Procter & Gamble Company

Characterization of the skin sensitisation potential is a key requirement for the safety assessment of cosmetic ingredients especially when significant dermal exposure to an ingredient is *** traditional methods to identify the skin sensitisation potential of chemicals have been guinea pig assays such as the Magnusson & Kligman or the Buehler *** a step forward,the mouse local lymph node assay（LLNA） has been implemented to replace the guinea pig *** LLNA is still an in vivomethod however provides significant scientific and animal welfare improvements such as quantitative and objective read-out,reduction in time and animal numbers *** underwent a thorough evaluation（OECD 429,442 A and B） and is widely applicable as the most technically sound in vivo test ***,nowadays the risk assessment for new ingredients must rely on non-animal methods as animal tests are no longer feasible in most instances,*** cosmetic *** recent years,significant progress has been made to develop non-animal methods to meet cosmetics animal testing ban *** alternative assays have been *** far three assays have been validated by OECD:（i） the direct peptide reactivity assay（DPRA,OECD TG 442 c）,（ii） the human cell line activation assay（h-CLAT,OECD TG 442 e）,（iii） the KeratinoSens assay（OECD TG 442 d）.In addition,a large number of other novel assays,have been developed by industry and ***,Cosmetic Europe（CE） funds an extensive research program,method development and method evaluation to refine the fundamental understanding of how key pathways contribute to the induction of skin sensitisation and develop in vitro test *** these alternative assays are aiming to address the individual key events in the skin sensitisation AOP（Adverse Outcome Pathway）.An objective framework is needed to interpret the results from these methods and their prediction models to facilitate decision making（hazard identification or safety

关键词： Alternative Methods for Skin Sensitisation Risk Assessment

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Pentachlorophenol cause spontaneous abortion through autophagy in pregnancy mice

Pentachlorophenol cause spontaneous abortion through autopha...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Xiaomin Huang Zhenyao Huang Xiumei Han Mingming Yu Yan Zhang Yun Fan Chuncheng Lu Yankai Xia State Key Laboratory of Reproductive Medicine Institute of ToxicologyNanjing Medical University Key Laboratory of Modern Toxicology of Ministry of Education School of Public HealthNanjing Medical University

Pentachlorophenol(PCP) is an environmental endocrine compounds usually used as herbicides and insecticides,which have carcinogenic,teratogenic,reproductive toxicity and is associated with spontaneous abortion in previous *** aim of this study was to evaluate the developmental toxicity of PCP by in vivo and in vitro experiments,and explored the mechanism of PCP leading to spontaneous *** ICR pregnancy mice were gavaged with PCP at forth dose(20μg/kg/bw/day,200μg/kg/bw/day,2 mg/kg/bw/day,20 mg/kg/bw/day) on embryonic days O.S to 8.5(GD0.5-8.5).The fetal lost rate on GD16.5 increased in a dose effect relationship in pregnancy *** HE staining showed that the sinusoidal structural is abnormal,epithelial cells reduced and trophoblast cells decreased in the *** autophagosome was observed in the electron microscopy(EM) in the PCP exposure *** RT-PCR and western blotting indicated that bcl2 decreased and beclinl enhanced while the LC3 II/LC3 I increased in placental in PCP exposure *** bioinformatic predictions,we found that miRNA-30 played an important role in autophagy gene *** RT-PCR of miRNA-30 expression was lower in placental in PCP exposure *** HTR-8/SVneo exposed to 20 uM and 200 Mm PCP to explore the *** autophagosome was also found in *** RT-PCR showed that the miRNA-30 was higher in PCP exposure cell,bcl2 reduced and beclinl *** gene test proved the combination of miRNA-30 and *** conclusion,PCP exposure can down-regulate the expression of miRNA30 which might mediate the autophagy through bcl2 reduction leading to increment of beclin1 to result in spontaneous abortion.

关键词： PCP Pentachlorophenol cause spontaneous abortion through autophagy in pregnancy mice

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Organic extracts of PM2.5 Repress Neuronal Differentiation from hESCs by Interfering GABA Signaling

Organic extracts of PM2.5 Repress Neuronal Differentiation f...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Wei Hongying Feng Yan Liang Fan Cheng Wei Zhou Ren Wang Yan The Ninth People Hospital of Shanghai Jiao Tong University School of Medicine School of Public Health Shanghai Jiao Tong University School of Medicine MOE-Shanghai Key Laboratory of Children's Environmental Health

Air pollution,especially fine particulate matter(PM),has been reported to be associated with neurodevelopmental *** potential mechanisms have been proposed to interpret the developmental neurotoxicity of ***,few studies focus on the relationship between signal transduction molecules and neuronal differentiation under ***,we aimed to investigate the effect of organic extracts of PM（Po） on neuronal differentiation from human embryonic stem cells（ESC） and the role of GABA signaling mechanisms during this *** H9 hES cell line expressed pluripotent markers OCT4,SOX2 and *** neural induction,nearly 98%cells form H9 were NESTIN and SOX2 dual-positive neural precursor cells（NPCs）.75%cells of NPCs could differentiate into neurons with typical neurite and characterized with higher levels of MAP2,TUJ1 and *** Po treatment,the differentiation states of neurons were significantly repressed,showing significant decreases in MAP2 staining,counts of newly differentiated neurons,neurite length and neurite branch ***,the mRNA level of GABRB3 was downregulated with a time-dependent manner under Po ***3 immunostaining also revealed obvious decreases in the distributions of GABRB3 on cell *** effects of GABRB3 agonist,GABRB3 antagonist or GABA confirmed the regulating role of GABRB3 in Po-induced impairments of neuronal *** downstream effects of GABA sinaling,BDNF and bHLH,also involved in the supression of neuronal differentiation induced by *** togther,our study demonstrated the involvement of GABA signaling dysfunction in Po-induced suppression in neuronal differentiation,uncovering an important molecular mechanism of GABA sinaling in the neurodevelopmental disorders associated with particulate air pollution.

关键词： Organic extracts of PM2.5 Repress Neuronal Differentiation from hESCs by Interfering GABA Signaling

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Acrylonitrile-induced mitochondrial dysfunction is correlated with CYP2E1 mitochondrial translocation in rats

Acrylonitrile-induced mitochondrial dysfunction is correlate...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： BoBo Yang Fang Liu Fang Li Ying Dong Suhua Wang Guangwei Xing Rongzhu Lu Department of Preventive Medicine and Public Health Laboratory Sciences School of MedicineJiangsu University

Cytochrome P450 2 E1(CYP2 E1) detoxifies or bioactivates many low molecular-weight *** knowledge about CYP2 E1 activity relies on studies of the enzyme localized to endoplasmic reticulum(ER) or exactly the microsomes(mcCYP2 E1);however,CYP2 E1 undergoes transport to mitochondria(mtCYP2 E1) and becomes metabolically *** present study reports the first pomparison of the expression and activity of hepatic mcCYP2 E1 and mtCYP2 E1 in rats exposed to acrylonitrile(AN).Male SD rats were randomly divided into four groups,i.e.,control,25 mg/kg AN,50 mg/kg AN and 75 mg/kg AN *** group was further divided into three subgroups depending on the time of treatment(1 h,6 h and24 h).The results showed AN significantly increased the expression and activity of mtCYP2 E1 and suppressed that of mcCYP2 E1,though the dose and time-responses were not consistent at all time ***,the total CYP2 E1 mRNA level was markedly reduced by AN at three dosages and time ***,AN reduced mitochondrial membrane potential(MMP),ATP and increased ROS in a dose-and time-dependent manner in *** conclusion,these results demonstrate that AN can promote the mitochondrial translocation of CYP2 E1,which may be resulted from the redistribution of CYP2 E1 in mitochondrial and ER,thus causing mitochondrial damages and increased mitochondrial ROS in rats.

关键词： mtCYP2E1 mcCYP2E1 acrylonitrile mitochondrial translocation mitochondrial damages

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Identification of Key Molecules Involved in Spatial Memory Impairment of Triple Transgenic Mice of Alzheimer's Disease Caused by Long-term Copper Exposure

Identification of Key Molecules Involved in Spatial Memory I...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Xifei Yang Jun Yu Xuemei Lin Jianjun Liu Key Laboratory of Modern Toxicology of Shenzhen Medical Key Laboratory of Guangdong ProvinceMedical Key Laboratory of Health Toxicology of ShenzhenShenzhen Center for Disease Control and Prevention

Alzheimer's disease(AD) is the most common neurodegenerative disease characterized by a progressive impairment of cognitive functions including spatial learning and *** copper exposure accelerates the development of AD;however,the potential mechanisms by which copper exacerbates the symptoms of AD remain *** this study,we explored the effects of chronic copper exposure on cognitive function by treating 6 month-old triple AD transgenic(3 xTg-AD) mice with 250 ppm copper sulfate in drinking water for 6 months,and identified several potential key molecules involved in the effects of chronic copper exposure on memory by proteomic *** behavioral test showed that chronic copper exposure aggravated memory impairment of 3 xTg-AD ***-dimensional fluorescence difference gel electrophoresis(2 D-DIGE) coupled with mass spectrometry revealed a total of 44 differentially expressed proteins(18 up-regulated and 26 down-regulated) in hippocampus between the wild-type(WT) mice and non-exposed 3 xTg-AD mice.A total of 40 differentially expressed proteins were revealed(20 up-regulated and 20 down-regulated) in hippocampus between copper exposed and non-exposed 3 xTg-AD *** these differentially expressed proteins,complexin-1 and complexin-2,two memory associated proteins,were significantly decreased in hippocampus of 3 xTg-AD mice compared with the WT ***,the expression of these two proteins was further down-regulated in 3 xTg-AD mice when exposed to *** abnormal expression of complexin-1 and complexin-2 identified by proteomic analysis was verified by Western-blot *** analysis revealed that overexpression of complexin-1 and complexin-2 prevented spatial impairment of 3 xTg-AD *** together,our data showed that chronic copper exposure accelerated memory impairment and altered the expression of proteins in hippocampus in3 xTg-AD *** functional analysis on the differentially expressed proteins suggested t

关键词： Alzheimer's disease Two-dimensional fluorescence difference gel electrophoresis Copper Memory impairment

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Could maternal urine estrogenic metabolite levels predict puberty timing in the offspring in rats after pregestational ethinyl estradiol exposure ?

Could maternal urine estrogenic metabolite levels predict pu...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Ningning Li Yaqi Li Kaixuan Xu Yankai Xia Di Wu State Key Laboratory of Reproductive Medicine Institute of ToxicologySchool of Public HealthNanjing Medical University Key Laboratory of Modern Toxicology of Ministry of Education School of Public HealthNanjing Medical University

Ethinyl estradiol(EE) is widely used in oral contraceptive *** tested whether metabonomics measuring urine estrogenic metabolites could be used to relate progestational EE exposure and puberty timing in the *** SD rats were gavaged with oil,50,200 or 800 μg/kg bw/day EE for 15 consecutive *** 15 days of washout,females mated and got *** samples were collected on the day before exposure,the 1 st and15 th day of washout,gestation(G) day 7,14 and *** estrogenic metabolite levels were measured using UPLC-MS/*** puberty timing were recorded and kisspeptin,GPR54,NK3 R,LeptinR and InsulinR protein levels in hypothalamic Arcuate(ARC) on Postnatal Day 60 were *** showed that progestational EE treatment could both significantly advance(800 μg/kg male) and delay(50 μg/kg male and 800μg/kg female) offspring puberty ***-treated groups had significantly higher level of ARC kisspeptin in female offspring and lower level in *** InsulinR level decreased significantly in females while ARC LeptinR level increased in both *** estrogenic metabolite analysis indicated that in male offspringpuberty timing were negatively associated with 2-MeOE2 on G7 and G14,4-MeOE2,Progesterone and 17α-OHprogesterone on G14,E1 on G21,butpositively associated with2-MeOE1 on *** females,puberty timing were positively associated with 2-MeOE1 on G14,but negatively associated with 16 a-OHE1 and 4-OHE1 on *** results indicated maternal estrogen metabolism data during middle and late pregnancy might help understand offspring puberty timing.

关键词： Pregestation Ethinyl estradiol Puberty timing Metabonomics Kisspeptin

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Overexpression of HO-1 assisted PM2.5-induced autophagy-related cell necrosis and apoptosis failure

Overexpression of HO-1 assisted PM2.5-induced autophagy-rela...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Wei Zhou Xiaoyan Yuan Li Zhang Baoting Su Dongdong Tian Yang Li Jun Zhao Sbuangqing Peng Evaluation and Research Center for Toxicology Institute of Disease Control and PreventionAcademy of Military Medical Sciences

Severe smog/haze events accompanied by extremely high concentrations of airborne fine particulate matter(PM2.5) are now a frequent occurrence in northern China and the potential health consequences are of growing *** these episodes,there is a reported surge in hospital visits for acute respiratory symptoms and respiratory diseases *** investigate the underlying pathogenic mechanisms,we exposed human lung epithelial cells(BEAS-2 B) to PM2.5(0,25,50,100,200,400 μg/mL) for 24 h and found that high doses of PM2.5 caused cell necrosis and autophagy dysfunction,while co-treatment with the autophagy inhibitor 3-MA could partially reduce PM2.5-induced cell *** to PM2.5 also increased the expression and mitochondrial transposition of heme oxygenase 1(HO-1),which consequently reduced the release of cytochrome c from mitochondria to *** of HO-1 by siRNA attenuated the mitochondrial accumulation of HO-1,reversed HO-1-dependent reduction of cytochrome c release and promoted PM2.5-induced cell *** contrast to necrosis,PM2.5-induced autophagy was independent of *** conclusion,our results demonstrate that acute exposure to high concentrations of PM2.5 causes autophagy-related cell necrosis,a cell death mode that increases local *** decrease in cytochrome c release and apoptosis by unregulation of HO-1 may be involved in PM2.5-induced autophagy-related cell ***,this study reveals dual roles for HO-1 in PM2.5-induced cytotoxicity and presents a possible explanation for the onset of acute respiratory symptoms under extreme particulate air pollution.

关键词： apoptosis Overexpression of HO-1 assisted PM2.5-induced autophagy-related cell necrosis and apoptosis failure

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PGC-1α介导的线粒体生物发生与分裂/融合的交互作用在鱼藤酮诱导的多巴胺神经元变性损伤中的作用研究

PGC-1α介导的线粒体生物发生与分裂/融合的交互作用在鱼藤酮诱导...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者：彭凯歌杨利葵肖瀞淞施锐曹佳赛燕第三军医大学军事预防医学系毒理学研究所

目的:鱼藤酮(rotenone)是一种从鱼藤及多种植物的根部提取的天然化合物,一直被视为安全有效的杀虫剂。大量的流行病学和实验室研究显示鱼藤酮与PD的发病相关。鱼藤酮为线粒体复合体I的抑制剂,线粒体在鱼藤酮所致的多巴胺神经元变性损伤... 详细信息

目的:鱼藤酮(rotenone)是一种从鱼藤及多种植物的根部提取的天然化合物,一直被视为安全有效的杀虫剂。大量的流行病学和实验室研究显示鱼藤酮与PD的发病相关。鱼藤酮为线粒体复合体I的抑制剂,线粒体在鱼藤酮所致的多巴胺神经元变性损伤中发挥了核心作用。因此,本课题通过建立鱼藤酮诱导的多巴胺神经元变性损伤细胞模型,探讨多巴胺神经元线粒体分裂/融合和生物发生在鱼藤酮致多巴胺神经元变形损伤过程中的交互作用及机制。方法:建立鱼藤酮诱导的高分化PC12细胞体外染毒模型,评价鱼藤酮对PC12细胞的毒性效应,并在透射电子显微镜下观察鱼藤酮对线粒体形态学、线粒体的长度和嵴的数量的影响;激光共聚焦显微镜下观察线粒体的片段化及线粒体质量的改变;同时,使用TMRM染料检测PC12细胞线粒体膜电位的改变。利用荧光实时定量PCR检测多巴胺神经元线粒体拷贝数;采用RT-PCR和WB实验检测线粒体生物发生相关因子PGC-1α和mtTFA,融合相关因子MFN2和OPA1,和分裂相关因子Drp1和Fis1基因转录和蛋白表达水平的表达变化;分别使用线粒体融合促进剂M1和分裂抑制剂Mdivi-1确定线粒体分裂/融合在鱼藤酮诱导的多巴胺神经元变性损伤中的作用;另外,通过siRNA转染和慢病毒感染构建低表达和过表达PGC-1α基因的细胞模型,确定PGC-1α对多巴胺神经元线粒体分裂/融合的调控作用。结果:鱼藤酮可以导致多巴胺神经元细胞活性降低,TH蛋白的表达下调,分泌释放多巴胺的能力降低;线粒体膜电位降低,活性氧的生成增加等。透射电镜下可见大量的小圈状或点状的碎片化线粒体,同时线粒体的长度、面积以及线粒体嵴的长度明显减小。同时,线粒体生物发生相关因子(PGC-1α和mtTFA)及线粒体融合分裂相关因子(MFN2、OPA1、Fis1)的基因转录和蛋白表达水平在鱼藤酮的作用下均出现不同程度的降低,而磷酸化Drp1蛋白却出现表达上升的趋势,与对照组比较具有显著性差异。线粒体分裂抑制剂M1和融合促进剂Mdivi-1可以提高细胞活性、增加线粒体DNA拷贝数以及TH蛋白的表达,减少线粒体DNA的片段化,具有改善的线粒体质量和形态的作用。过表达PGC-1α可显著抑制鱼藤酮诱导的PC12细胞的死亡,提高线粒体质量和线粒体DNA拷贝数,升高了MFN2的蛋白表达并显著降低了p-Drp1的蛋白表达水平,因此,过表达PGC-1α对鱼藤酮诱导的多巴胺神经毒性有显著的保护作用。而低表达PGC-1α可加剧鱼藤酮诱导的PC12细胞的神经毒性作用,MFN2的蛋白表达水平显著下降,p-Drp1的蛋白表达水平显著上升。另外,免疫荧光实验的结果也进一步提示提示鱼藤酮可促使p-Drp1蛋白表达升高并从胞浆向线粒体转位。结论:鱼藤酮暴露可以引起多巴胺神经元线粒体生物发生障碍、分裂/融合失衡进而导致多巴胺神经元线粒体形态结构和功能的异常。PGC-1α通过调控线粒体生物发生与分裂/融合的交互作用在鱼藤酮诱导的多巴胺神经元的变性损伤中发挥了重要作用。

关键词：鱼藤酮线粒体生物发生分裂/融合 PGC-1α 多巴胺神经元

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Silica nanoparticles induce abnormal mitosis and apoptosis via PKC-δmediated negative signaling pathways in GC-2 cells of mice

Silica nanoparticles induce abnormal mitosis and apoptosis v...

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2017（第三届）毒性测试替代方法与转化毒理学（国际）学术研讨会

作者： Jin Zhang Lihua Ren Jialiu Wei Jianhui Liu Yanbo Li Caixia Guo Zhixiong Shi Zhiwei Sun Xianqing Zhou Department of Toxicology and Hygienic Chemistry School of Public HealthCapital Medical University Beijing Key Laboratory of Environmental Toxicology Capital Medical University

Backgrounds The potential health hazards of silica nanoparticles(SiNPs) have attracted more and more *** had shown that SiNPs could damage seminiferous epithelium and reduce the quantity and quality of sperms,but little is known about the toxicity mechanism of SiNPs on male reproductive *** present study was designed to investigate toxic mechanism of SiNPs on male mice using spermatocyte lines(GC-2 spd cells),with exposure to 0,6.25,12.5,25 and 50 μg/mL SiNPs for 24 h,***:SiNPs could enter GC-2 cells and mainly localized in the cytoplasm and *** SiNPs damaged mitochondria structures such as swollen mitochondria,mitochondrial crest fracture and vacuolization in ***,SiNPs induced a dose-dependent reduction in cell viability,as well as increases in the LDH release and cell apoptosis rate in GC-2 *** could lead to DNA strand breaks and abnormal mitosis,arresting GC-2 cells at the G0/G1 ***,SiNPs could simultaneously activate PKC-8 mediated negative signaling pathway(PKC-δ/p53/p21 cip1) and PKC-α mediated positive signaling pathway(PKC-α/MAPK).However the lower expressions of cyclin E and cyclin-dependent kinases 2(CDK2) indicated PKC-δsignaling pathway played a major role in the process of cell ***:The present results suggest that SiNPs can be internalized into GC-2 cells,then disturb PKC mediated signaling pathway of cell cycle through DNA strand breaks resulting from oxidative *** PKC-δmediated negative signaling play a leading *** activation of PKC-δ signaling pathway led to cell cycle arrest and apoptosis,resulting in abnormal *** present study may provide new evidence to elucidate the toxicity mechanisms of male reproductive system and may be beneficial for safety assessment of SiNPs products.

关键词： Silica nanoparticles GC-2spd cell Mitosis Cell cycle PKC signaling pathway

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