三疣梭子蟹ftz-f1 基因的克隆及相关核受体基因在蜕皮中的功能分析

采用RACE 技术克隆三疣梭子蟹核受体fzt-f1 基因, 全长cDNA 序列1763bp.该基因3'和 5'非编码区分别为1034bp 和141bp, 开放阅读框为588bp, 推测编码195 个氨基酸, 预测分子量为 22.8kDa, 理论等电点为6.35, 命名为PTNHr 基因。同源性和系统进化分析表明, 三疣梭子蟹fzt-f1 基因与刀额新对虾同源性高达75.4%, 并且与刀额新对虾聚为一支。对核受体基因fzt-f1、EcR 和RXR 在蜕皮周期中的表达情况进行实时荧光定量PCR 分析, 结果表明, 三个核受体基因在不同蜕皮时期, 均出现表达差异, 说明这三个基因都参与蜕皮调控过程。其中, fzt-f1 和RXR 基因在不同蜕皮时期的 表达模式上出现相反的表达特征, 预测这两个基因存在相互抑制的调节关系。EcR 和RXR 基因在不 同蜕皮时期的表达模式上出现部分相似的表达特征。     

半胱胺对斜带石斑鱼（Epinephelus coioides）生长激素及其受体mRNA水平的影响

以海水硬骨鱼类斜带石斑鱼为研究对象,通过投喂和腹腔注射半胱胺盐酸盐（CSH）,研究半胱胺对斜带石斑鱼生长激素（GH）及其受体（GHR）mRNA水平的影响,以初步揭示半胱胺促进斜带石斑鱼生长的作用机理。结果表明,长期投喂CSH可以显著提高斜带石斑鱼的相对体重增长率。投喂和腹腔注射不同剂量的CSH均能明显促进斜带石斑鱼脑垂...     

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。     

Endocrine effects of methoxylated brominated diphenyl ethers in three in vitro models

Methoxylated brominated diphenyl ethers (MeO-BDEs) in aquatic environments have been found to be primarily of natural origin in the marine environment and not from biotransformation of synthetic PBDEs. Two of the eight MeO-PBDEs (2′-MeO-BDE-68 and 6-MeO-BDE-47) that were detected in anchovy from the Yangtze River Delta, were natural products from marine organisms. So 2′-MeO-BDE-68 and 6-MeO-BDE-47 were chosen to study the potential to modulate androgen, estrogen, or thyroid hormone receptor- (AR, ER, ThR) mediated responses by use of reporter gene assays. 2′-MeO-BDE-68 was antiandrogenic at 50 μM, estrogenic at 10 μM and antiestrogenic at 10 and 50 μM (IC₅₀ = 4.88 μM). 2′-MeO-BDE-68 enhanced luciferase expression by 5 nM T3 at 50 μM. 6-MeO-BDE-47 exhibited potent antiandrogenicity at 1 μM and greater (IC₅₀ = 41.8 μM) and possessed estrogenic activity at 10 μM and antiestrogenic activity at 10 and 50 μM (IC₅₀ = 6.02 μM).     

Cell bioassays for detection of aryl hydrocarbon (AhR) and estrogen receptor (ER) mediated activity in environmental samples

In vitro cell bioassays are useful techniques for the determination of receptor-mediated activities in environmental samples containing complex mixtures of contaminants. The cell bioassays determine contamination by pollutants that act through specific modes of action. This article presents strategies for the evaluation of aryl hydrocarbon receptor (hereafter referred as dioxin-like) or estrogen receptor mediated activities of potential endocrine disrupting compounds in complex environmental mixtures. Extracts from various types of environmental or food matrices can be tested by this technique to evaluate their 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents or estrogenic equivalents and to identify contaminated samples that need further investigation using resource-intensive instrumental analyses. Fractionation of sample extracts exhibiting significant activities, and subsequent reanalysis with the bioassays can identify important classes of contaminants that are responsible for the observed activity. Effect-directed chemical analysis is performed only for the active fractions to determine the responsible compounds. Potency-balance estimates of all major compounds contributing to the observed effects can be calculated to determine if all of the activity has been identified, and to assess the potential for interactions such as synergism or antagonism among contaminants present in the complex mixtures. The bioassay approach is an efficient (fast and cost effective) screening system to identify the samples of interest and to provide basic information for further analysis and risk evaluation.     

Modulation by Biogenic Amines for the Hemocyte Count and Prophenoloxidase Exocytosis via Receptors in Litopenaeus vannamei

Hemocyte counts and phenoloxidase(PO)activity were examined after hemolymph being incubated in dopamine(DA),noradrenaline(NE)and serotonin(5-HT).Results showed that all the three biogenic amines(BAs)had a significant impact on total hemocyte count(THC),differential hemocyte count(DHC),and intracelluar and extracelluar phenoloxidase(PO)activity.Among these Bas,DA had the strongest effect on the above parameters,whereas 5-HT had the least effect.Preincubation with D1 receptor antagonist SCH23390,D2 receptor a...     

Removal of Natural Free Estrogens and their Conjugates in a Municipal Wastewater Treatment Plant

Removal of natural free estrogens and estrogen conjugates in a municipal wastewater treatment plant (WWTP) was investigated and analyzed by GC‐MS, in which estrogen conjugates were first transformed to their corresponding free estrogens with an acid solvolysis procedure before their analysis. Natural free estrogens, E1‐3‐sulfate (E1‐3S), and E3‐3‐sulfate (E3‐3S) were detected with high concentrations in both the influent and effluent of the primary settling tank (PS), while no estrogen glucuronides were detected in any of the monitored wastewater samples. Regarding their removal efficiencies, all were almost completely removed, except for E1 with only a minor decrease. The estrogenic/androgenic removal of the same WWTP was also evaluated with estrogen receptor (androgen receptor) (ER (AR))‐binding assays, in which the removal efficiencies for E2 equivalents (EEQ) or testosterone equivalents (TEQ) were 68.5 and 72.2%. In addition, the chemically calculated EEQ from natural estrogens were about 20.6–39.3% that of the ER‐binding assay, in which E3 contributed the biggest proportion in both the influent and PS, while the calculated value of E1 increased from only 6.7% in the influent to as high as 20.6% in the effluent.     

Rainfall–runoff modelling of ephemeral streams in the Valencia region (eastern Spain)

This paper presents preliminary results from the application of a transfer‐function rainfall–runoff model to ephemeral streams in Mediterranean Spain. Flow simulations have been conducted for two small catchments (Carraixet and Poyo basins), located in close proximity to one another yet with significantly different geological characteristics. Analysis of flow simulations for a number of high‐flow events has revealed the dominant influence of the rainfall on the catchment response, particularly for high‐rainfall events. Particular success has been attained modelling the highest magnitude events in both catchments and for all events in the faster responding (Poyo) catchment. In order to investigate the viability of the model for forecasting floods in ungauged catchments, additional investigations have been conducted by calibrating the model for one catchment (donor catchment) and then applying it to another (receptor catchment). The results indicate that this can be successful when either the donor catchment is a fast response catchment or when the model is calibrated using a high‐magnitude event in the donor catchment, providing that the modelled receptor catchment event is of a lower magnitude. Copyright © 2002 John Wiley & Sons, Ltd.     

许氏平鲉(Sebastes schlegelii)卵子发生及妊娠过程中雌激素、芳香化酶及其受体的变化规律

雌二醇是鱼类卵巢分泌的主要的性类固醇激素之一,在脊椎动物卵子发生、性别分化、卵黄形成、生理免疫以及中枢神经系统调节等生理过程中发挥重要的作用。许氏平鲉营胎生繁殖,且经过漫长的卵黄积累期和妊娠期,这一特殊的生殖方式使得它受到了国内外学者的广泛关注。通过分析与雌二醇合成及作用密切相关的cyp19a和Era在许氏平鲉卵巢上的定位、cyp19a和cyp19b mRNA在不同卵巢发育时期及不同组织中的表达以及不同发育期的卵巢中雌二醇含量变化,发现在卵巢发育的过程中,许氏平鲉体内雌二醇含量的峰值出现在IV期和体节期。并且cyp19a和cyp19b在卵巢中均有表达且在不同的卵巢发育期表达量不同。在卵黄积累的过程中, cyp19a和cyp19b在IV期的表达量最高,在妊娠时体节期表达量最高。不仅如此,在V期卵巢的滤泡层、基质和囊胚期的滤泡胎盘上检测到了强烈的Era的信号。这些结果说明雌二醇在胎生鱼类卵子发生及妊娠过程中均起到非常重要作用。这一研究为深入解析许氏平鲉胎生生殖特性积累了资料。