Primary neuronal-astrocytic co-culture platform for neurotoxicity assessment of di-(2-ethylhexyl) phthalate

Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.     

焦亚硫酸钠对大鼠背根节神经元钾电流的影响

应用全细胞膜片钳技术,研究SO2体内衍生物和食品添加剂———焦亚硫酸钠(sodiummetabisulfite,SMB)对大鼠背根节(DRG)神经元瞬间外向钾电流(TOCs)和延迟整流钾电流(IK)的影响.结果表明,SMB可增大TOCs和IK,且具剂量依赖性和电压依赖性.10μmolL-1的SMB不影响TOCs的激活过程,给药前后TOCs的半数激活电压为(3.15±2.29)mV和(4.72±1.92)mV(N=8,P>0.05),不改变其斜率;但10μmolL-1的SMB却非常显著地影响TOCs的失活过程,给药前后其半数失活电压分别为(-53.6±0.45)mV和(-38.85±0.68)mV(N=8,P<0.01),也不改变其斜率.10μmolL-1的SMB显著提前IK的激活过程,给药前后IK的半数激活电压为(-11.98±3.50)mV和(-33.21±5.67)mV(N=8,P<0.01),不改变其斜率.SMB显著增大TOCs和IK,使IK的激活过程提前,抑制TOCs的失活过程,从而导致细胞内K 通过K 通道外流增加,进而可能对DRG神经元功能产生不利影响.图6参16     

Adverse effects of metal exposure on chemotaxis towards water-soluble attractants regulated mainly by ASE sensory neuron in nematode Caenorhabditis elegans

Chemotaxis to water-soluble attractants is mainly controlled by ASE sensory neuron whose specification is regulated by che-1 in Caenorhabditis elegans. Our data suggested that exposure to high concentrations of metals, such as Pb, Cu, Ag, and Cr, would result in severe defects of chemotaxis to water-soluble attractants of NaCl, cAMP, and biotin. Moreover, the morphology of ASE neuron structures as observed by relative fluorescent intensities and relative size of fluorescent puncta of cell bodies, relative lengths of sensory endings in ASE neurons, and the expression patterns of che-1 were obviously altered in metal exposed animals when they meanwhile exhibited obvious chemotaxis defects to water-soluble attractants. In addition, the dendrite morphology could be noticeably changed in animals exposed to 150 mol/L of Pb, Cu, and Ag. Furthermore, we observed significant decreases of chemotaxis to water-soluble attractants in Pb exposed che-1 mutant at concentrations more than 2.5 mol/L, and in Cu, Ag, and Cr exposed che-1 mutant at concentrations more than 50 mol/L. Therefore, impairment of the ASE neuron structures and functions may largely contribute to the appearance of chemotaxis defects to water-soluble attractants in metal exposed nematodes.     

Brief exposure to copper induces apoptosis and alters mediators of olfactory signal transduction in coho salmon

Pacific salmon are particularly susceptible to copper (Cu)-induced olfactory injuries that can ultimately inhibit neurobehaviors critical to survival. However, the molecular mechanisms underlying Cu-mediated olfactory impairment remain poorly understood. In the present study, we conducted a short-term Cu exposure at levels relevant to urban runoff (5, 25 and 50 ppb) , and investigated the roles of impaired olfactory signal transduction and induced apoptosis as underlying mechanisms of olfactory injury. Increased cell death in the olfactory epithelium was evident in coho receiving 4 h exposures to 25 and 50 ppb Cu. Expression of olfactory marker protein (omp), a marker of mature olfactory sensory neurons, also decreased at 50 ppb Cu. Immunohistochemical analysis of coho olfactory epithelium demonstrated a loss of type 3 adenylate cyclase (ACIII) in the apical olfactory epithelium cilia at all levels of Cu exposure, suggesting an inhibitory effect of Cu in olfactory signaling. Accompanying the loss of ACIII in Cu-exposed coho were reduced intracellular cyclic guanosine monophosphate (cGMP) levels in the olfactory rosettes. Collectively, these results support a linkage among the initial steps of olfactory signaling in Cu-induced salmon olfactory injury, and suggesting that monitoring olfactory cGMP levels may aid in the assessment of salmon olfactory injury.     

Mother gestational exposure to organophosphorus pesticide induces neuron and glia loss in daughter adult brain

Chlorpyrifos (CPF) is a widely used organophosphorus pesticide with developmental neurotoxicity such as morphogenesis toxicity. In the present study, we assessed the effects of prenatal CPF exposure on systemic parameters and cytoarchitecture of medial prefrontal cortex (mPFC) in adulthood. Gestational dams were exposed to 5mg/kg/d of CPF during gestational days 13–17, while body weight, organ coefficient, and neuron and glia counts of offspring were determined on postnatal day 60. Our results showed that CPF treatment induced little or no effects on body weight and organ coefficients. There were also no significant pathological changes in mPFC. However, neuron and glia count analysis showed that CPF treatment reduced neuron and glia counts in anterior cingulate, prelimbic, and infralimbic areas of mPFC. The CPF react pattern was similar in both sexes, and there was no statistical difference in most of the sub-regions. Thus, our results revealed an embryonic origin brain deficit induced by gestational mother pesticide exposure.     

SO2衍生物对大鼠神经元和心肌细胞几种离子通道的影响

为了阐明大气污染物SO2对神经系统和心血管系统的毒作用机制,采用全细胞膜片钳技术研究了SO2衍生物(NaHSO3和Na2SO3,分子比为1∶3)对大鼠海马、背根节神经元和心肌细胞膜上钠、钾、钙离子通道的影响.结果显示:(1)SO2衍生物可显著增大大鼠海马CA1区神经元钠电流,不影响钠通道的激活过程,但可使钠电流的失活曲线向去极化方向移动,延迟钠通道的失活过程;另外,SO2衍生物可显著增大瞬间外向钾电流(IA)和延迟整流钾电流(IK),不影响IA的激活过程,使IK的激活过程向负电压方向移动,促进IK的激活过程,而使IA的失活曲线向正电压方向移动,延迟IA的失活过程.(2)SO2衍生物显著增大大鼠背根节神经元钠电流(TTX-S钠电流和TTX-R钠电流),可使两种钠电流的激活和失活曲线均向去极化方向移动,但对失活的影响大于对激活的影响,即延迟钠通道的失活过程;SO2衍生物显著增大背根节神经元瞬间外向钾电流(TOCs)和延迟整流钾电流(IK),不影响TOCs的激活过程,但可使IK的激活曲线向超极化方向移动,促进IK的激活.另外,还可使TOCs的失活曲线向去极化方向移动,即延迟TOCs的失活.(3)SO2衍生物可显著增大大鼠心肌细胞L-型钙电流(ICa,L),使ICa,L的激活和失活曲线均向去极化方向移动,但对失活的影响大于对激活的影响;SO2衍生物显著增大心肌细胞钠电流,不影响钠通道的激活过程,但可使钠电流的失活曲线向去极化方向移动,延迟钠通道的失活过程;SO2衍生物显著增大心肌细胞瞬间外向钾电流(Ito),使Ito的激活曲线向超极化方向移动,促进Ito的激活过程,但可使Ito的失活曲线向去极化方向移动,延迟Ito的失活过程;此外,SO2衍生物还可显著增大心肌细胞内向整流钾电流(IK1),但不影响其反转电位.结果表明,SO2衍生物可能通过影响神经元和心肌细胞膜上离子通道的活动而对中枢神经、传导神经以及心血管系统产生不利影响.提示大气SO2污染可能与神经系统和心血管系统疾病的发生有关.     

海域水质模型参数动态反演方法研究初探

海域水质模型长周期数值模拟中,模型参数全时段统一赋值的方法忽略了参数随时间动态变化的物理特性,降低了模型的可靠性,增加了海域水质模型验证工作的难度.本文建立了将数据驱动模型和水质模型有机结合的参数动态反演的新方法:以水质模型多参数设计工况的数值模拟,构建海域内部观测点污染物浓度响应解集,并将解集划分为若干时段;应用基于人工神经网络的数据驱动模型归纳建立观测点每一时段内污染物浓度同多个模型参数之间的非线性关系;将实测资料带入关系中,进行模型参数随时间变化的动态反演.以渤海海域水质模型为例,采用“孪生”实验验证参数动态反演新方法的可行性,结果表明该方法是有效的,能够保证模拟周期内较高的数值精度,提高了模型的准确性.     

不同季节大气可吸入颗粒物(PM_(10))诱导小鼠原代培养神经元炎性及其级联损伤效应

为了探讨不同季节可吸入颗粒物(PM10)对神经元的损伤效应,本研究通过建立小鼠大脑皮层原代神经元体外染毒模型,考察了不同季节PM10对炎性因子诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)和黏附分子(ICAM-1)表达水平的影响,并探讨了与之耦联的磷酸化Ca2+/钙调蛋白依赖性蛋白激酶Ⅱα(p-Ca MKⅡα)、环磷腺苷效应元件结合蛋白(p-CREB)及即早基因(c-jun、c-fos)的表达情况.结果表明,PM10暴露刺激神经元炎性细胞因子释放增加,显著上调p-Ca MKⅡα和p-CREB的水平,并刺激即早基因的高表达,且以冬季PM10的效应最为明显.由此提示,PM10暴露可能通过炎症反应机制激活Ca2+-Ca MK-CREB通路,进而刺激即早基因表达引发神经毒性作用,而冬季PM10中多环芳烃类物质负荷可能是造成效应季节性差异的主要原因.     

基于RS-ANN的通风系统可靠性预警系统

系统可靠性预警是度量系统运行状态偏离可靠性指标界线的强弱程度,确定预警等级和做出决策警示的过程。笔者在对目前国内外有关系统预警方法的分析比较基础上,针对矿井通风系统可靠性运行的实际状况,应用了粗糙集(RS)理论和神经网络(ANN)技术,提出了一种基于粗糙集神经网络(RSANN)的矿井通风系统可靠性预警方法:首先,建立了一套适合于矿井通风系统可靠性的预警指标体系;然后,利用人工神经网络与粗糙集理论的优势互补,以粗糙集作为前置处理系统优化指标结构,构建了基于RSANN的通风系统可靠性预警仿真模型,并应用该模型进行了实例验证。其结果表明,该模型的仿真结论与基于ANN的结论十分吻合,训练效率提高了667倍。     

基于自适应神经网络的通风系统故障过程及其规律研究

首先对影响通风系统故障过程特征量进行了定义;然后通过对矿井通风系统故障过程及其改善和劣化状态的分析,建立了基于两参数的Weibull过程故障率函数,给出了系统故障过程改善和劣化的判别式;最后,基于自适应神经网络技术对通风系统的故障规律进行了仿真研究,并应用C 语言和Matlab语言强大的解算及绘图功能,求出了故障过程特征参数值,绘制了故障曲线.