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.     

非节肢类海洋无脊椎动物中蜕皮甾体及其功能研究

非节肢类海洋无脊椎动物的许多物种中都发现有蜕皮甾体的存在,且存在形式多样,但目前对其确切来源、生物体内的合成或加工途径仍缺乏足够的了解,在生长发育、生殖活动及其他生理功能中如何发挥作用也需要进一步研究。分子生物学和全基因组测序的发展为蜕皮甾体合成途径重要酶类及蜕皮激素信号传导所需受体研究提供了更多的信息,以帮助科学家逐步阐明相关机制。     

Analysis of IL-2-1ike factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder,Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2, (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%. This work was supported by National “973” Project G1999012003, G19999012006.     

Identification of a novel C-type lectin from the shrimp Litopenaeus vannamei and its role in defense against pathogens infection

Acting as one of the pattern recognition receptors (PRRs), C-type lectin is believed to mediate pathogen recognition and plays an important role in the clearance of pathogens as part of the innate immune system. In this work, a novel C-type lectin gene (named LvLec1) was cloned from the shrimp Litopenaeus vannamei. The ORF of LvLec1 is 510 bp, encoding 169 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids at the N-terminal and a carbohydrate recognition domain...     

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。     

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.     

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。     

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。     

Array technology as a tool to monitor exposure of fish to xenoestrogens

A variety of anthropogenic chemicals are capable of binding to the estrogen receptor of vertebrate species. Binding of these compounds can interfere with homeostasis by disrupting normal gene expression patterns. The purpose of this study was to investigate the feasibility of applying array technology as a monitoring tool for detecting the presence and distribution of estrogenic compounds in coastal habitats using sheepshead minnows as our model. cDNA clones that were isolated from differential display, including vitellogenin alpha and beta, vitelline envelope protein (ZP2), and transferrin, among others, were spotted on the macroarray. The results of these experiments demonstrate a characteristic expression pattern of estrogen responsive genes in sheepshead minnows exposed to 17 beta-estradiol (E2).