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研究了用重组孕激素受体基因酵母测定饮用水中内分泌干扰物的方法,并利用该种方法检测了南方某水厂不同处理工艺过程水样对孕激素受体活性的抑制水平.结果表明,重组孕激素受体基因酵母能够与孕激素专一性的结合,诱导产生明显的剂量-效应关系,EC50值为0.5 nmol/L,具有较高灵敏度;环境内分泌干扰物五氯酚和壬基酚具有孕激素受体抑制活性,其IC50值分别为2.4μmol/L和3.7μmol/L;重组孕激素受体基因/报道基因的酵母技术是一种筛选和定量分析具有孕激素受体抑制活性的内分泌干扰物的快速、有效方法.结合固相萃取的前处理技术,重组孕激素受体基因酵母对水厂不同处理工艺水样检测出明显的孕激素受体抑制活性,抑制率均大于58%,表明重组孕激素受体基因酵母检测技术能够快速监测和鉴别水样中具有抑制孕激素受体活性的物质. 相似文献
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利用固相萃取-高效液相色谱法调查了松花江流域某自来水厂原水、传统净水工艺各处理单元出水及管网水中的13种内分泌干扰物(EDCs).结果表明,固醇类雌激素物质除管网水中未检出外,其他各水样均有检出,浓度为4-44 ng·L-1;4种邻苯二甲酸酯(PAEs)、双酚A(BPA)及3种烷基酚(APs)在各水样中几乎100%检出,浓度为2~163 760 ng·L-1,其中以邻苯二甲酸二丁酯(DBP)与邻苯二甲酸二(2-乙基己基)酯(DEHP)为主.研究还发现,尽管有个别EDCs出水浓度高于进水浓度,但混凝沉淀工艺对EDCs的去除在该水厂中占主导作用,平均去除率为63%;砂滤与氯消毒工艺出水不稳定,只能对部分EDCs起到一定程度去除作用.此外,出厂水经管网输送后其中的绝大多数EDCs含量升高,说明进入管网水中的EDCs除水源水污染外,还有一个重要途径就是输水管材中相应污染物的渗析. 相似文献
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对国内的饮用水水源地内分泌干扰物污染现状进行总结,发现我国饮用水水源地很多地区都存在着内分泌干扰物的污染,由于地域的不同,污染物的种类存在差异,但有机氯农药和多氯联苯类化合物的污染尤为严重。同时对水源地内分泌干扰物的种类、来源及生物学效应进行综述,并对内分泌干扰物的毒性作用机制进行了归纳,发现不同来源的内分泌干扰物,毒性作用机制不同,且一种内分泌干扰物可能同时存在多种作用机制。 相似文献
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重组基因酵母筛选法因其快速灵敏、廉价易行和高通量而成为较为广泛应用的内分泌干扰物筛选法之一.采用96孔培养板替代三角锥瓶对酵母筛选法进行了改进.改进后的方法更加简便,节省了大量试剂. 相似文献
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使用固相萃取-酵母双杂交法研究了东北地区某污水厂内分泌干扰活性随季节的变化及不同处理单元的去除情况.结果表明,该法的EC50值为5.95×10-11mol·L-1,具有较高灵敏度.对污水水样检测得出,原污水及一沉池出水内分泌干扰活性雌二醇当量值分别为16.13~21.65和17.10~22.85n·gL-1,均以夏季最高、冬季最低;经过A/O工艺处理,二沉池出水的内分泌干扰活性降至3.37~6.76n·gL-1,以冬春较高、夏秋较低;整个污水处理工艺对内分泌干扰活性的去除率为58.1%~84.2%,平均为75.2%,也以夏季最高、冬季最低.研究还发现,二级生物处理工艺对内分泌干扰活性起主导去除作用,而一沉池对内分泌干扰物的去除能力非常有限,甚至较进水有升高现象发生. 相似文献
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介绍了发光细菌法、重组基因酵母法、免疫分析法、细胞增殖法、受体结合活性法、卵黄蛋白法、酵母双杂交法等几种环境中内分泌干扰物的生物检测技术及研究进展,并提出了展望。 相似文献
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环境内分泌干扰物前处理方法研究新进展 总被引:1,自引:0,他引:1
环境内分泌干扰物因其能够紊乱生物体正常内分泌功能而引起人们的广泛关注,然而内分泌干扰物在水环境中浓度极低,水体中复杂机质的干扰较大,在分析监测前一般要经过样品前处理过程,以起到富集痕量组分、消除基体干扰、提高方法灵敏度的作用。针对不同环境样品中内分泌干扰物的前处理方法不同,分别介绍了液体、固体或类固体环境样品的前处理技术,提出了各前处理方法的优缺点,总结并展望了环境内分泌干扰物样品前处理方法的发展方向。 相似文献
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环境内分泌干扰物浅析 总被引:5,自引:0,他引:5
国内外对环境内分泌干扰物的研究证实,人类的生殖障碍、发育异常、某些癌症及某些免疫系统、神经系统疾病与环境内分泌干扰物的效应有关,本文对环境内分泌干扰物概念及问题产生背景、种类、对人类健康的危害及作用机制作了浅要的综合分析,并提出了对污染现状的认识与防治对策。 相似文献
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接有芳香烃受体(AhR)基因和芳香烃受体核转位子蛋白(ARNT)基因的酵母Saccharomyces cerevisiae菌株是测定化学物质的Ah受体效应的方法之一.本实验通过优化各个实验条件,发现用0.2%葡萄糖作前培养液培养菌24 h,用2%半乳糖作化学物质暴露时的培养液,暴露在玻璃管时,暴露时间从原来的18 h缩短到8 h.并用已经优化的方法测定了六氯苯和五氯苯的Ah受体效应,得出六氯苯和五氯苯相对于TCDD的毒性等当量值(TEF)分别为0.018629和0.000294. 相似文献
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Weiwei Jiang Ye Yan Mei M Donghong Wang Qian Luo Zijian Wang Senthil Kumaran Satyanarayanan 《环境科学学报(英文版)》2012,24(2):320-328
Detection of estrogenic disrupting compounds (EDCs) in drinking waters around China has led to rising concerns about health risks associated with these compounds. There is, however, a paucity of studies on the occurrence and identification of the main compounds responsible for this pollution in the source waters. To fill this void, we screened estrogenic activities of 23 source water samples from six main river systems in China, using a recombinant two-hybrid yeast assay. All sample extracts induced significant estrogenic activity, with E2 equivalents (EEQ) of raw water ranging from 0.08 to 2.40 ng/L. Additionally, 16 samples were selected for chemical analysis by gas chromatography-mass spectrometry. The EDCs of most concern, including estrone (E1), 17βup-estradiol (E2), 17αup-ethinylestradiol (EE2), estriol (E3), diethylstilbestrol (DES), estradiol valerate (EV), 4-t-octylphenol (4-t-OP), 4-nonylphenols (4-NP) and bisphenol A (BPA), were determined at concentrations of up to 2.98, 1.07, 2.67, 4.37, 2.52, 1.96, 89.52, 280.19 and 710.65 ng/L, respectively. Causality analysis, involving comparison of EEQ values from yeast assay and chemical analysis identified E2, EE2 and 4-NP as the main responsible compounds, accounting for the whole estrogenic activities (39.74% to 96.68%). The proposed approach using both chemical analysis and yeast assay could be used for the identification and evaluation of EDCs in source waters of China. 相似文献
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Studies on estrogenic disrupting compounds(EDCs) occurrence and identification of main responsible compounds in river water discharged into the sea are of significance.In the present research,we screened estrogenic activities of 10 river water samples from 3 main rivers discharged into Bohai Sea in Tianjin using a recombinant two-hybrid yeast assay and chemical analysis by gas chromatography-mass spectrometry.All sample extracts induced significant estrogenic activity,with 17β-estradiol equivalents(EEQ) of raw water ranging from 5.72 to 59.06 ng/L.Six most concerned EDCs in the river water samples including estrone,17β-estradiol,17α-ethinylestradiol,estriol,diethylstilbestrol and estradiol valerate were determined,with their concentrations up to 50.70,31.40,24.40,37.20,2.56,and 8.47 ng/L,respectively.Through causality analysis by comparing the EEQ values of yeast assay and chemical analysis,17α-ethinylestradiol and 17β-estradiol were identified as the main contributors to the estrogenic effects of the river samples,accounting for the whole estrogenic activities(62.99% to 185.66%),and estrogen antagonistic compounds might presented in the heavy polluted water samples.The proposed approach using both chemical analysis and bioassay could be used for identification and evaluation of the estrogenic activity of EDCs in river water. 相似文献
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Na Li Weiwei Jiang Kaifeng Rao Mei M Zijian Wang Satyanarayanan Senthik Kumaran 《环境科学学报(英文版)》2011,23(2):301-306
Environmental chemicals in drinking water can impact human health through nuclear receptors. Additionally, estrogen-relatedreceptors (ERRs) are vulnerable to endocrine-disrupting e ects. To date, however, ERR disruption of drinking water potency hasnot been reported. We used ERR two-hybrid yeast assay to screen ERR disrupting activities in a drinking water treatment plant(DWTP) located in north China and in source water from a reservoir, focusing on agonistic, antagonistic, and inverse agonistic activityto 4-hydroxytamoxifen (4-OHT). Water treatment processes in the DWTP consisted of pre-chlorination, coagulation, coal and sandfiltration, activated carbon filtration, and secondary chlorination processes. Samples were extracted by solid phase extraction. Resultsshowed that ERR antagonistic activities were found in all sample extracts, but agonistic and inverse agonistic activity to 4-OHTwas not found. When calibrated with the toxic equivalent of 4-OHT, antagonistic e uent e ects ranged from 3.4 to 33.1 g/L. In thetreatment processes, secondary chlorination was e ective in removing ERR antagonists, but the coagulation process led to significantlyincreased ERR antagonistic activity. The drinking water treatment processes removed 73.5% of ERR antagonists. To our knowledge,the occurrence of ERR disruption activities on source and drinking water in vitro had not been reported previously. It is vital, therefore,to increase our understanding of ERR disrupting activities in drinking water. 相似文献
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It is generally known that there are many endocrine disrupting compounds (EDCs) in the e uents from wastewater treatmentplants (WWTPs). Most research has focused on the occurrence of estrogenic or androgenic activities, while ignoring that there areenvironmental chemicals disrupting thyroid system, which is essential for growth and development in both humans and animals. Inthe present work, a two-hybrid yeast assay was conducted to evaluate the removal e ciencies of agonistic or antagonistic thyroidreceptor (TR) mediated e ects in di erent treatment processes of three WWTPs located in Beijing. We found no TR agonistic, but TRantagonistic activities in all processes from the WWTPs. The TR antagonistic activities in organic extracts of water samples were thencalibrated regarding to a known TR-inhibitor, amiodarone hydrochloride (AH). The observed concentration of TR disrupting substancesranged from 2.35 10??8 to 6.19 10??7 mol/L AH in Gaobeidian WWTP, 3.76 10??8 to 8.75 10??8 mol/L AH in Lugouqiao WWTP,and 4.80 10??9 to 2.55 10??8 mol/L AH in Beixiaohe WWTP. Of the three WWTPs, the removal rates were 92.7%, 42.2%, and 23.1%respectively. Industrial sewage may contain more TR disrupting substances compared with domestic sewage. The recipient waters werefound to contain considerable concentrations of TR disrupting substances that may cause adverse e ects on the exposed organisms. 相似文献
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It is generally known that there are many endocrine disrupting compounds (EDCs) in the effluents from wastewater treatment plants (WWTPs). Most research has focused on the occurrence of estrogenic or androgenic activities, while ignoring that there are environmental chemicals disrupting thyroid system, which is essential for growth and development in both humans and animals. In the present work, a two-hybrid yeast assay was conducted to evaluate the removal efficiencies of agonistic or antagonistic thyroid receptor (TR) mediated effects in different treatment processes of three WWTPs located in Beijing. We found no TR agonistic, but TR antagonistic activities in all processes from the WWTPs. The TR antagonistic activities in organic extracts of water samples were then calibrated regarding to a known TR-inhibitor, amiodarone hydrochloride (AH). The observed concentration of TR disrupting substances ranged from 2.35 × 10-8 to 6.19 × 10-7 mol/L AH in Gaobeidian WWTP, 3.76 × 10-s to 8.75 x 10-s mol/L AH in Lugouqiao WWTP,and 4.80 × 10-9 to 2.55 × 10-s mol/L AH in Beixiaohe WWTP. Of the three WWTPs, the removal rates were 92.7%, 42.2%, and 23.1% respectively. Industrial sewage may contain more TR disrupting substances compared with domestic sewage. The recipient waters were found to contain considerable concentrations of TR disrupting substances that may cause adverse effects on the exposed organisms. 相似文献
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酵母双杂交技术构建重组雌激素受体相关受体(ERR)基因酵母 总被引:1,自引:0,他引:1
应用酵母双杂交技术构建了重组雌激素受体相关受体(ERR)基因酵母,用以筛选环境中具有ERR干扰活性的化合物.实验先提取并纯化含有ERRγ基因的酵母表达质粒pGBT9-ERRγ,称为诱饵质粒;再提取并纯化含有ERRγ协同激活因子GRIP1基因的酵母表达质粒,形成靶质粒pGAD424-GRIP1,然后将诱饵质粒和靶质粒同时转化到酵母细胞Y187中,于营养缺陷型培养基(SD/-Trp/-Leu)上筛选阳性菌落,构建ERRγ-GRIP1双杂交酵母.同时,考察了ERRγ-GRIP1酵母与典型的雌激素拮抗剂——4-羟基他莫西酚(4-OHT)的结合情况.结果表明,4-OHT能够抑制ERRγ-GRIP1酵母酶活性,并存在显著的剂量-效应关系,半数抑制浓度(RIC50)值为7.9×10-6mol·L-1.最后,进一步建立了检测环境化合物诱导/抑制ERRγ-GRIP1酵母β-半乳糖苷酶活性的实验方法,用于检测典型的内分泌干扰物.双酚A(BPA)实验结果表明,BPA不能诱导/抑制ERRγ介导的酶活性,但BPA与4-OHT共同暴露时,能够修复4-OHT的抑制活性;经体外代谢活化后,BPA的修复活性得到加强,证实了该重组基因酵母用于环境污染物内分泌干扰效应筛选的可行性. 相似文献