Towards molecular understanding of species differences in dioxin sensitivity: initial characterization of Ah receptor cDNAs in birds and an amphibian

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related planar halogenated aromatic hydrocarbons (PHAHs) are highly toxic to most vertebrate animals, but there are dramatic species differences in sensitivity, both within and among vertebrate classes. For example, studies in cultured avian hepatocytes have revealed differential sensitivity of birds to PHAHs [Kennedy et al. (1996). Toxicol. Appl. Pharmacol., 141, 214-230]. Differences in the characteristics or expression of the aryl hydrocarbon receptor (AHR) could contribute to these species differences in PHAH responsiveness. To investigate the molecular mechanism of differential PHAH sensitivity, we have begun to characterize the AHR in white leghorn chicken (Gallus gallus), Pekin duck (Anas platyrhynchos), and common tern (Sterna hirundo), as well as an amphibian, mudpuppy (Necturus maculosus). Partial AHR cDNAs encompassing the helix-loop-helix and PAS domains were cloned and sequenced. Comparison of amino acid sequences in this region indicated a high degree of sequence conservation among the bird species (97% amino acid identity). The percent identity between bird sequences and either mouse or mudpuppy was lower (79%); the mudpuppy AHR was 74% identical to the mouse AHR. Phylogenetic analysis of these and other AHR amino acid sequences showed that the bird and mudpuppy AHRs were more closely related to mammalian and fish AHR1 forms than to fish AHR2. Future studies include the in vitro expression and functional characterization of AHRs from these and other non-mammalian vertebrates.     

盐度变化对黑鲷生长激素及其受体的影响

将海水鱼类黑鲷Sparusmacrocephalus从盐度为26.6的海水直接转入淡水,考察渗透压的急剧降低对黑鲷的生长激素(growthhormone,GH)及其受体(growthhormonereceptor,GHR)等生理指标的影响。研究发现:(1)黑鲷从海水直接转入淡水后48h有33%的鱼死亡,对照组无死亡现象;(2)黑鲷在转入淡水48h内其血清生长激素(GH)含量、肝脏及鳃中GHR及GHRmRNA含量与对照组均无显著差异;(3)黑鲷在转入淡水48h后其肾脏中GHR含量较对照组显著升高(p<0.05),肾脏中GHRmRNA含量在24h及48h显著高于对照组(p<0.05)。结果表明,从GH含量看,黑鲷从海水环境到淡水低渗环境与鲑鳟鱼类从淡水到海水环境的适应性调节机制有差异,而黑鲷肾脏中GHR的含量变化可能参与黑鲷对外界环境渗透压的这种急剧降低的适应性调节。     

基于客观赋权法的长江流域环境污染事故风险受体脆弱性时空变异特征研究

从敏感性和恢复力两方面描述了环境风险受体脆弱性,综合考虑人群、社会经济和生态环境系统等风险受体的敏感性和恢复力,选取12个指标构建区域环境风险受体综合脆弱性评价指标体系。以长江流域为研究对象,引入"纵横向-拉开档次"法和"时序加权平均算子"法等客观赋权法探讨了流域脆弱性时空变异特征。结果表明,2000-2011年长江流域环境风险受体脆弱性呈波动下降趋势,且脆弱性空间差异较大,其中,上海、河南、江苏和西藏等省区脆弱性指数最高,陕西、重庆和云南等省市脆弱性较低。研究结果可为长江流域环境风险管理和污染事故应急管理提供依据。     

Influence of plant on the extractability of estrogen in the rhizospheric soil

Livestock wastes applied to agricultural land are potential sources of steroidal hormones （estradiol （E2）, estrone （E1）, and estriol （E3）） that can adversely affect the aquatic ecosystem as endocrine disruptors. But the effects of plants on the fate of estrogen in the environment are not clearly known. In the present report, the behavior of E1, E2 and E3 added to the rhizospheric soil in various concentrations and its effect on plant growth were examined by estrogen batch sorption test, plant pot test and estrogen extraction test. Trifolium repens was grown on the soil spiked with E2 or a mixture of E1, E2 and E3. Pots without plant were prepared as controls. All pots were watered in 50 ml dose almost every day, and the rhizosheric soil and plant body were collected 7, 19, 26, 33 days after planting. The soil was sequentially extracted with Milli-Q water, and methanol / 1M acetic acid solvent followed by methanol wash at the solid to liquid ratio of 1 ： 4 （g/mL）. Extracts were analyzed for El, E2 and E3 using a Shimadzu GCMS-QP5050. Plant dry weight （4 hours, 105 ℃） was measured as an indicator of plant growth. In batch sorption tests, E1 and E2 concentrations in the liquid phase significantly decreased within 24 hours after the addition of estrogen. The sum of methanol / 1M acetic acid extractable E 1 and E2 in the soil decreased to half of the initial added amount within 7 days. Reduction in total estrogenic activity of the soil extracts （estimated as 0.01CE1＋CE2＋0.1CE3 where CE1, CE2 and CE3 are respective El, E2 and E3 concentrations per unit soil weight） continued for 1 month due to slow oxidation of E2 to E1. Desorption of E2 from soil during the pot experiment was higher than expected from the sorption experiment.     

Estrogen receptor (ER) agonists and androgen receptor (AR) antagonists in effluents from Norwegian North Sea oil production platforms

The in vitro estrogen receptor (ER) agonist and androgen receptor (AR) antagonist potencies of offshore produced water effluents collected from the Norwegian Sector were determined using recombinant yeast estrogen and androgen screens. Solid phase extraction (SPE) concentrates of the effluents showed E2 agonist activities similar to those previously reported for the United Kingdom (UK) Continental Shelf (<0.1-4 ng E2 L(-1)). No activity was detected in the filtered oil droplets suggesting that produced water ER activity is primarily associated with the dissolved phase. Targeted analysis for methyl- to nonyl-substituted alkylphenol isomers show the occurrence of known ER agonists in the analysed samples. For the first time, AR antagonists were detected in both the dissolved and oil associated phase at concentrations of between 20 and 8000 microg of flutamide equivalents L(-1). The identity of the AR antagonists is unknown, however this represents a significant input into the marine environment of unknown compounds that exert a known biological effect. It is recommended that further analysis using techniques such as bioassay-directed analysis is performed to identify the compounds/groups of compounds that are responsible in order to improve the assessment of the risk posed by produced water discharges to the marine environment.     

扇贝(Chlamys farreri)多肽通过EGFR抑制受紫外线B(UVB)诱导的HaCaT细胞凋亡

利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。     

扫描电镜分析在大气颗粒物源解析中的应用

扫描电镜法作为大气颗粒物源解析的辅助方法,为源解析研究提供了定性和半定量的依据。运用扫描电镜法对吉林市采暖期和非采暖期的大气颗粒物样品以及源样品进行定性和半定量分析,将分析结果与CMB8.2化学质量平衡受体模型源解析结果进行比较,两者具有一致性。吉林市采暖期对TSP贡献较大的是扬尘和土壤风沙尘,非采暖期对TSP贡献较大的是土壤风沙尘和建筑尘。     

副溶血性弧菌产耐热直接溶血毒素对黑色素瘤的抑制作用研究

为探讨耐热直接溶血毒素(Thermostabile direct hemolysin,TDH)在体内和体外对小鼠黑色素瘤细胞B16的抑制作用,本研究通过MTT法、克隆形成试验、凋亡试验、Caspase-8和Caspase-3的活性试验、线粒体膜电位的检测以及体内C57BL/6小鼠(Mus musculus)荷瘤实验,比较TDH作用于不同细胞的半抑制浓度(IC₅₀),评价TDH对小鼠黑色素瘤细胞B16的体内外抑制作用。结果发现:人结肠上皮细胞NCM460、人正常肝细胞LO2、人肝癌细胞SMMC-7721和小鼠黑色素瘤细胞B16在TDH处理24 h之后,细胞的半抑制质量浓度IC₅₀分别为151、118、54和48 μg/mL,正常细胞的IC₅₀高出癌细胞近2~3倍。当质量浓度低于20 μg/mL时,TDH以剂量依赖性的方式抑制B16细胞的克隆形成,6 mg/kg TDH在移植瘤模型中显著抑制体内肿瘤的生长(P<0.001)。流式细胞术和荧光试剂盒检测表明:20 μg/mL的TDH能诱导19.4%的B16细胞发生早期凋亡,并激活Caspase-8和Caspase-3,但不影响线粒体膜电位。TDH具有体内外的抗肿瘤活性,可能通过细胞表面的死亡受体介导的凋亡信号通路引起凋亡,从而发挥抗肿瘤作用。