不同终点检测5种双酚A类化合物对MCF-7的细胞毒性

双酚A(BPA)及其类似物作为聚碳酸酯的主要合成原料,一直是环境污染的重要问题.其雌激素效应是当今科学研究的重点,而毒性效应研究甚少.为评估环境中BPA类物质的毒性效应,实验采用四唑盐(MTS)比色法检测5种双酚A类化合物对人雌激素受体缺失乳腺癌细胞MCF-7(ER-)增殖活性的影响,2,4-二硝基苯肼法检测细胞乳酸脱氢酶(LDH)露出率,单细胞凝胶电泳(SCGE)检测DNA损伤.用非线性最小二乘法对MTS实验结果拟合剂量-效应曲线,表明所有的剂量-效应曲线(DRC)均能用Weibull或者Logit函数有效表征.以模型估算的半数效应浓度负对数值(pEC50)评估5种化合物的毒性大小依次为:BPB>BPC>TDP>BPE>BPA.LDH检测以及SCGE检测受试化合物对MCF-7(ER-)的损伤作用表明,在效应浓度EC20下,细胞核DNA轻微损伤,细胞增殖受轻微抑制;在效应浓度EC40下,细胞核DNA损伤严重,细胞增殖受到显著抑制,从而导致细胞膜通透性显著改变,使LDH大量露出.     

固定化果胶酶抑制铜绿微囊藻生长研究

为证实固定化果胶酶抑制蓝藻生长的作用,在实验室条件下,以铜绿微囊藻（Microcystis aeruginosa）为受试藻种,用共培养法观察了固定化果胶酶对藻细胞群体的作用、用电镜观察了共培养后藻细胞的损伤状况,测定了对其生理生化特征的影响.结果表明固定化果胶酶与藻共培养液第3 d明显黄化,且黄化程度与固定化果胶酶的用量和培养时间呈正相关系;电镜照片显示固定化果胶酶对藻细胞有损伤作用,轻微损伤的藻细胞出现质壁分离,表面粗糙、凸凹不平,形状不规则,严重损伤的藻细胞表面发生深度皱缩或细胞结构完全解体;随着固定化果胶酶与铜绿微囊藻共培养时间的延长,藻细胞生长量、叶绿素a含量显著降低,表明藻细胞受到胁迫和伤害,藻细胞正常的光合作用受到严重影响.丙二醛（MDA）值显示藻细胞抗氧化防御体系被破坏,细胞内发生严重膜脂过氧化.固定化果胶酶能有效抑制铜绿微囊藻细胞的生长,铜绿微囊藻生长抑制率可高达96%.     

NO参与铝诱导蚕豆保卫细胞死亡的调控

铝(Al)是地壳中含量最丰富的金属元素,是酸性土壤中导致植物生长抑制和作物产量下降的一个主要因素,但铝毒性作用机制尚不清楚.本文以蚕豆叶表皮为材料,研究铝胁迫对气孔保卫细胞活性的影响,探讨NO在铝诱导细胞死亡中的作用.结果表明,一定浓度的A1Cl3可诱导气孔保卫细胞活性降低,部分细胞死亡,且随着铝浓度的增高细胞死亡率增高.死细胞呈现核固缩、核崩解、凋亡小体等典型凋亡特征,且凋亡抑制剂Z-Asp-CH2-DCB能阻止AlCl3诱发的细胞死亡.用NO清除剂c-PTIO、NO合酶抑制剂L-NAME或硝酸还原酶抑制剂NaN3降低铝处理组胞内NO后,细胞死亡率显著降低,胞内ROS、Ca2+水平同期降低;NaN3还能降低铝处理组中具有程序性死亡特征的细胞比率.用ROS清除剂AsA清除铝处理组胞内ROS后,细胞死亡率显著降低,胞内Ca2+和NO水平亦显著降低;铝处理液中加入Ca2+通道抑制剂LaCl3后,细胞死亡率低于铝单独处理组,胞内ROS和NO水平无明显改变.研究结果表明,铝胁迫引起的胞内NO合成增加通过Ca2+信号途径介导了保卫细胞的程序性死亡.     

小飞扬草(Euphorbia thymifolia L.)中镉的亚细胞分布及化学形态

采用差速离心法和逐步提取法,分析Cd超积累植物小飞扬草根、茎、叶中Cd的亚细胞分布和化学形态。培养液中Cd浓度5mg/L和10mg/L条件下,在小飞扬草根、茎、叶中,细胞壁和细胞膜是Cd主要的结合场所,含量分别占总量的47.1%～49.0%、39.7%～41.4%、42.9%～56.2%,其次是胞液组分,含量分别占37.5%～41.4%、28.2%～40.7%、35.2%～46.8%,细胞器组分中Cd含量较少。Cd在小飞扬草根、茎、叶中均以氯化钠提取态为主,含量分别占70.5%～84.8%、72.4%～83.0%、50.0%～74.8%。根部各提取态含量依次为CNaCl>CHAC>CEtOH>CHCl>CW>CR,茎、叶中各提取态含量依次为CNaCl>CHCl>CHAC>CEtOH>CW>CR。在小飞扬草中,Cd与细胞壁和细胞膜中的果胶酸和蛋白质等物质结合固定,而进入细胞的Cd大部分与有机酸络合而隔离于液泡内,这可能是小飞扬草忍耐并超积累Cd的机制。     

邻苯二甲酸二丁酯暴露对雄性大鼠生精细胞功能影响

观察不同剂量的邻苯二甲酸二丁酯(DBP)对成年大鼠生精细胞损伤、端粒酶和端粒酶逆转录酶(TERT)表达影响.对40只健康雄性SD大鼠随机分成4组,分别为500mg·kg-1·d-1、250mg·kg-1·d-1、50mg·kg-1·d-1 3个染毒组和1个对照组,灌胃量为5mL·kg-1体重,灌胃法连续给药6周.对体重...     

Competition of bloom-forming marine phytoplankton at low nutrient concentrations

Competition of three bloom-forming marine phytoplankton (diatom Skeletonema costatum, and dinoflagellates Prorocentrum minimum and Alexandrium tamarense) was studied through a series of multispecies cultures with di erent nitrate (NaNO3) and phosphate (NaH2PO4) levels and excess silicate to interpret red tide algae succession. S. costatum outgrew the other two dinoflagellates in nitrate and phosphate replete cultures with 10 mol/L Na2SiO3. Under nitrate limited (8.82 mol/L NaNO3) conditions, the growth of S. costatum was also dominant when phosphate concentrations were from 3.6 to 108 mol/L. Cell density of the two dinoflagellates only increased slightly, to less than 400 and 600 cells/mL, respectively. Cell density of S. costatum decreased with time before day 12, and then increased to 4000 cells/mL (1.5 mg/L dry biomass) at NaNO3 concentrations between 88.2 and 882 mol/L with limited phosphate (0.36 mol/L NaH2PO4) levels. In addition, P. minimum grew well with a maximal cell density of 1690–2100 cells/mL (0.5–0.6 mg/L dry biomass). Although S. costatum initially grew fast, its cell density decreased quickly with time later in the growth phase and the two dinoflagellates were dominant under the nitrate-limited and high nitrate conditions with limited phosphate. These results indicated that the diatom was a poor competitor compared to the two dinoflagellates under limited phosphate; however, it grew well under limited nitrate when growth of the dinoflagellates was near detection limits.     

培育和壮大知识产权产业的市场主体

党的十六届三中全会《关于完善社会主义市场经济体制若干问题的决定》指出,要“全面推进经济法制建设。按照依法治国的基本方略,着眼于确立制度、规范权责、保障权益”。这一重要论述对完善我国市场主体制度和现代产权制度具有重要指导作用。国家正在完善各类市场主体的法律制度,使各类市场主体真正具有完全的行为能力和责任能力。     

环讯瞭望

北京 环保部发布指导意见 培育引导环保社会组织发展 从环境保护部获悉：为促进环保社会组织健康、有序发展,努力构建全民参与环保的社会行动体系,环境保护部近日出台《关于培育引导环保社会组织有序发展的指导意见》,明确了培育引导环保社会组织的基本原则和总体目标。     

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.     

Mutagenic and cytotoxic potential of Endosulfan and Lambda-cyhalothrin - In vitro study describing individual and combined effects of pesticides

Excessive use of pesticides poses increased risks to non target species including humans. In the developing countries, lack of proper awareness about the toxic potential of pesticides makes the farmer more vulnerable to pesticide linked toxicities, which could lead to diverse pathological conditions. The toxic potential of a pesticide could be determined by their ability to induce genetic mutations and cytotoxicity. Hence, determination of genetic mutation and cytotoxicity of each pesticide is unavoidable to legislate health and safety appraisal about pesticides. The objective of current investigation was to determine the genotoxic and cytotoxic potential of Endosulfan（EN） and Lambda-cyhalothrin（LC）; individually and in combination. 3-（4,5-Dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium Bromide（MTT） assay was utilized to determine cytotoxicity, while two mutant histidine dependent Salmonella strains（TA98, TA100） were used to determine the mutagenicity of EN and LC.Moreover, mutagenicity assay was conducted with and without S9 to evaluate the effects of metabolic activation on mutagenicity. Even though a dose dependent increase in the number of revertant colonies was detected with EN against both bacterial strains, a highly significant（p 〈 0.05） increase in the mutagenicity was detected in TA98 with S9. In comparison, data obtained from LC revealed less mutagenic potential than EN. Surprisingly,the non-mutagenic individual-concentrations of EN and LC showed dose dependent mutagenicity when combined. Combination of EN and LC synergistically induced mutagenicity both in TA98 and TA100. MTT assay spotlighted comparable dose dependent cytotoxicity effects of both pesticides. Interestingly, the combination of EN and LC produced increased reversion and cytotoxicity at lower doses as compared to each pesticide, concluding that pesticide exposure even at sub-lethal doses can produce cytotoxicity and genetic mutations, which could lead to carcinogenicity.