Molecular characterization and developmental expression patterns of thyroid hormone receptors (TRs) and their responsiveness to TR agonist and antagonist in Rana nigromaculata

Considering some advantages of Rana nigromaculata as an experimental species, we propose that this species, like Xenopus laevis, could be used to assay thyroid hormone（TH） signaling disrupting actions. To validate the utilizability of R. nigromaculata, we investigated the responsiveness of R. nigromaculata to a TH receptor（TR） agonist（T3） and antagonist（amiodarone） by analyzing expression, based on characterizing TR cDNA and developmental expression patterns. With high levels of identity with the corresponding genes in X. laevis, both TRα and TRβ in R. nigromaculata exhibited roughly similar developmental expression patterns to those of X. laevis, in spite of some species-specific differences. Both TRα and TRβ expression had greater changes in the liver and intestine than in the tail and brain during metamorphosis. T3 exposure for 2 days induced more dramatic increases of TRβ expression in stage 27 than in stage34 tadpoles but not in stage 42 tadpoles, showing that the responsiveness of R. nigromaculata to TH decreased with development and disappeared at the onset of metamorphic climax.Corresponding to greater changes of TRβ expression in the liver and intestine than in the tail and brain during metamorphosis, the liver and intestine had higher responsiveness to exogenous T3 than the tail and brain. Amiodarone inhibited T3-induced TRβ expression. Our results show that R. nigromaculata can be used as a model species for assaying TH signaling disrupting actions by analyzing TRβ expression, and intestine tissues at stage 27 are ideal test materials due to high responsiveness and easy accessibility.     

A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone (TH) signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A (TBBPA), a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3, whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.     

植物激素对平菇菌丝生长及酶活性的影响

采用固体平板和液体摇瓶培养方法,研究了赤霉素、2,4-D、吲哚乙酸和萘乙酸4种激素对平菇-126菌丝生长及酶活性的影响。结果表明,吲哚乙酸虽然能提高发酵液胞外蛋白的含量和过氧化物酶活性,但对菌丝的生长起抑制作用,这与吲哚乙酸能引起平菇菌丝裂解和细胞膜渗漏有关;萘乙酸、低浓度2,4-D能促进菌丝的伸长;萘乙酸、赤霉素在低浓度下也能促进菌丝的分枝,对菌丝的生长整体上起促进作用。除吲哚乙酸外,其他3种激素对平菇菌丝生长影响都有两重性：低浓度下能促进菌丝的生长,浓度过高则会对菌丝生长产生毒害作用。     

Re-evaluation of thyroid hormone signaling antagonism of tetrabromobisphenol A for validating the T3-induced Xenopus metamorphosis assay

We developed the T3-induced Xenopus metamorphosis assay, which is supposed to be able to sensitively detect thyroid hormone(TH) signaling disruption of chemicals. The present study aimed to validate the T3-induced Xenopus metamorphosis assay by re-evaluating the TH signaling antagonism of tetrabromobisphenol A(TBBPA), a known TH signaling disruptor. According to the assay we developed, Xenopus tadpoles at stage 52 were exposed to 10–500 nmol/L TBBPA in the presence of 1 nmol/L T3. After 96 hr of exposure, TBBPA in the range of 10–500 nmol/L was found to significantly inhibit T3-induced morphological changes of Xenopus tadpoles in a concentration-dependent manner in term of body weight and four morphological endpoints including head area(HA), mouth width(MW), unilateral brain width/brain length(ULBW/BL), and hind-limb length/snout-vent length(HLL/SVL).The results show that these endpoints we developed are sensitive for characterizing the antagonistic effects of TBBPA on T3-induced metamorphosis. Following a 24-hr exposure,we found that TBBPA antagonized expression of T3-induced TH-response genes in the tail,which is consistent with previous findings in the intestine. We propose that the tail can be used as an alternative tissue to the intestine for examining molecular endpoints for evaluating TH signaling disruption. In conclusion, our results demonstrate that the T3-induced Xenopus metamorphosis assay we developed is an ideal in vivo assay for detecting TH signaling disruption.     

生产源区人血中多溴联苯醚水平与甲状腺激素相关性研究

采用气相色谱/负化学电离源-质谱法对莱州湾南岸地区十溴联苯醚生产源区36位居民血清中的5种多溴联苯醚浓度进行了检测,同时测定了样品中的甲状腺激素水平.血清中Σ5PBDEs（BDE-28、-47、-153、-183、-209）的质量浓度范围（以脂重计算,下同）为130.3～4 478.4 ng.g-1,平均值为529.9 ng.g-1.5种同族体中,BDE-209对Σ5PBDEs的平均贡献率达到了69.8%.在与甲状腺激素水平的相关性研究中,计算了两者的Spearman秩相关系数,结果显示BDE-28、-47、-153、-183与TSH水平呈高度负相关,BDE-183与fT4水平呈现显著性负相关,而BDE-28、-47与T3,BDE-28、-153、-183与fT3之间均呈现显著性正相关.本研究中,生产源区人体血清中PBDEs水平处于较高的暴露水平,主要同类物为BDE-209,人体对PBDEs的暴露会影响上述4种甲状腺激素的水平,PBDEs对人体的影响不容忽视,有关PBDEs暴露水平与甲状腺激素水平之间的相关性,应进一步深入研究.     

氧化处理水中典型酚类污染物研究进展

环境激素是一类能干扰人类和动物正常内分泌的外源性物质,对生物体的生理过程产生很大的危害。本文介绍了水处理中,典型酚类污染物氧化剂降解技术的机理、特点、应用以及局限性,主要介绍了Fenton试剂、O3、二氧化氯等氧化剂及高铁酸钾新型氧化剂在酚类废水处理中的应用。并指出了复合氧化剂、新型氧化剂与其他水处理技术的联合应用将是酚类废水氧化处理的发展方向。     

北京市工业废水和城市污水中菊酯类农药残留分析

建立了基于18C固相萃取柱和气相色谱/电子捕获（GC/ECD）分析水体中环境激素类物质--菊酯类农药的分析方法,并对方法的回收率、灵敏度进行了评价,同时分析了北京市七类典型污染点源50个采样点位菊酯类农药的浓度,检出的菊酯类农药包括联苯菊酯、氰戊菊酯、氯氰菊酯和溴氰菊酯,其检出率都较低,低于22%.检出菊酯类农药的浓度范围是0.013～1.246μg/L.该方法对菊酯类农药的回收率达到67.7%～96.2%,检测限为0.010～0.015μg/L.溴氰菊酯在废水排放中的达标率为100%.     

环境激素双酚A释放及降解进展

通过对环境激素双酚A的概念、危害、应用、释放、降解与去除的介绍，指出在今后的科研工作中，加强双酚A降解菌的筛选，并研究其生理、生化、遗传特性，对从分子水平了解其降解机制以及构建高效工程菌具有重要意义。     

A Screening Test for the Determination of Ethylene Sensitivity

Ethephon, which releases ethylene within plant tissues after application, was chosen to perform assessments of the relative sensitivity of crops to ethylene and to determine which stages of plant development were most sensitive. The species chosen were: barley, wheat, oats, canola and field pea, all of which are important crops in the province of Alberta, Canada. Plants were treated with ethephon at one of 7 different stages. Plants were assessed for their vegetative and reproductive growth, including height, biomass, yield and seed quality. Visual symptoms were photographed and documented to compare them with symptoms caused by ethylene applied as a gas. It was concluded that in barley, wheat and canola the late vegetative and early reproductive stages were most sensitive, at least when sensitivity was defined as reductions in yield and quality. As for field pea, ethephon had no effect on yield but did cause increased numbers of pods, which in certain conditions could lead to increased yields. Significant effects on vegetative growth were only observed in the early vegetative stages of development but with no effects on yield. The screening protocol successfully identified sensitive cultivars and growth stages for further investigation of the effects of ethylene exposure.