运用爪蟾变态实验检测镉的甲状腺激素干扰效应

将NF51阶段非洲爪蟾(Xenopus laevis)蝌蚪暴露于Cd2+浓度为0.01,0.1和1mg.L-1的溶液中21d,观测Cd2+对爪蟾的生长、变态和甲状腺组织结构的影响.1mg.L-1Cd2+暴露组爪蟾蝌蚪与对照组相比,7d后,平均体重减少了0.39倍,体长和后肢长分别减少了0.17倍和0.27倍,发育阶段稍有延缓;21d后,1mg.L-1Cd2+暴露组爪蟾蝌蚪后肢长减小了0.11倍,延缓了1个发育阶段.其余各浓度组在7d和21d暴露后,体重、体长、后肢长和发育阶段与对照组无显著差异.21d后,Cd2+各浓度组甲状腺组织结构出现滤泡数目减少、胶质减少甚至空泡化等现象,1mg.L-1Cd2+暴露组甲状腺滤泡之间还出现溶通现象.结果表明CdCl2具有甲状腺激素干扰效应,并可能是由于该化合物直接作用于甲状腺组织引起.由此可见,爪蟾变态实验可以有效检测重金属盐的甲状腺激素干扰效应.     

全氟辛磺酸对于爪蟾的甲状腺激素干扰效应

运用两栖类变态实验(AMA)研究了全氟辛磺酸(PFOS)对于非洲爪蟾(Xenopus laevis)蝌蚪的生长、变态和甲状腺组织结构的影响.结果显示,与对照组相比,7d后,10,50和250μg.L-1PFOS暴露组蝌蚪的后肢长分别减小了17%,11%和18%,蝌蚪的发育延缓了1~2个阶段;19d后,10,50和250μg.L-1PFOS组蝌蚪的体重分别增加了17%,10%和20%,体长分别增加了5%,4%和6%,而10μg.L-1和250μg.L-1PFOS组蝌蚪的后肢长分别减小了12%和13%;暴露组甲状腺组织结构出现滤泡数目减少、上皮细胞增生、胶质减少甚至滤泡溶通等现象.结论是,在本实验浓度下,PFOS对于非洲爪蟾的蝌蚪生长有促进作用,对于变态发育过程却有抑制作用.参照AMA的方法,可以判断PFOS为抗甲状腺激素干扰物.     

热带爪蟾胚胎对三丁基锡致畸作用响应的敏感时期

采用200 ng·L-1氯化三丁基锡对热带爪蟾(Xenopus tropicalis)胚胎进行第1天、第2天、第3天和3d全暴露实验.结果表明,第3天和3d全暴露组胚胎的存活率比对照组分别降低了10％和20％,第2天、第3天和3d全暴露组的体长分别减少了30％、20％和39％.暴露组胚胎畸形率均在90％以上,畸形类型主...     

视黄酸对热带爪蟾胚胎致畸效应的表型特征

取典型的胚胎致畸剂——全反式视黄酸对热带爪蟾(Xenopus tropicalis)胚胎进行24,36和48 h暴露.结果表明,2,10和50 μg·L^-1视黄酸暴露对胚胎的存活率没有影响,对胚胎的生长和发育却有明显的抑制作用,并导致所有胚胎出现畸形现象,说明视黄酸对热带爪蟾胚胎具有极强的致畸性,同时显示了热带爪蟾胚胎对视黄酸的敏感性.视黄酸主要引起胚胎脑部缩小、眼睛色素减少、围心腔水肿和尾巴弯曲等多种畸形类型. 24,36和48 h后,暴露组脑部畸形指数和眼睛畸形指数较对照组均有明显增加,并表现出较好的浓度-效应关系,而时间-效应关系不明显,表明视黄酸对脑和眼部的影响主要发生在暴露的前24小时;48 h后尾部的畸形指数相对于24 h有明显的升高.将视黄酸导致的胚胎畸形类型与污染物三丁基锡对比,表明视黄酸致畸特征图谱在三丁基锡致畸机制的研究中有重要的参考价值.由此可见,有可能将FETAX实验发展为一种基于致畸机制的发育毒性检测方法.     

三丁基锡暴露对褐菖鲉肾脏抗氧化防御系统的影响

观察TBT(1、10、100 ng.L-1)对褐菖鲉(Sebastiscus marmoratus)肾脏的抗氧化防御系统的影响.在实验条件下,分别于暴露后7、25、50 d和恢复实验的7、20 d采样进行理化分析.结果显示褐菖鲉暴露于TBT之后,肾脏MDA含量显著升高,之后MDA水平恢复到与对照组相同的水平;GST活性基本上无明显变化;TBT的暴露导致GPx活性产生诱导升高;对GSH含量影响的除7d时1 ng.L-1组显著减少,在暴露期间基本上没有变化.结果说明,褐菖鲉肾脏抗氧化防御系统对环境水平的TBT暴露是敏感的,而肾脏存在着有效的机制抵御氧化胁迫.     

三丁基锡（TBT）对褐菖鲉肝脏抗氧化系统的影响

使用环境浓度水平[1、10、100 ng(Sn)/L]的三丁基锡(tributyltin,TBT)对褐菖鲉进行水体暴露,观察TBT对褐菖鲉肝脏抗氧化防御系统的影响.暴露50 d后,10、100 ng/L组丙二醛含量的上升指示了环境浓度水平的TBT能够导致肝脏氧化胁迫.10、100 ng/L组的谷胱甘肽过氧化酶、超氧化物歧化酶和10 ng/L组的过氧化氢酶活性在TBT暴露50 d后被显著诱导.还原型谷胱甘肽含量在10、100 ng/L TBT暴露7 d后被显著诱导,而在暴露50 d后被浓度依赖性地抑制.TBT暴露7 d后,谷胱甘肽硫转移酶在10、100 ng/L组被显著诱导,而100 ng/L组在暴露25 d后被抑制.在恢复期7和20 d后,各浓度组的这些生物指标都回到与对照组相当的水平.本研究的结果表明:环境浓度水平的TBT能够引起鱼体肝脏氧化胁迫,这些生物标志物能够指示海洋环境中TBT的污染.     

三苯基锡对热带爪蟾胚胎的早期发育毒性

有机锡的外暴露能引起脊椎动物胚胎的独特畸形,但通过母体传递引起的内暴露毒性还缺乏研究.本文通过显微注射的方式研究了三苯基锡(TPT)、过氧化物酶体增殖体激活受体γ(PPARγ)的激动剂罗格列酮(Rosi)和抑制剂T0070907(C_(12)H_8ClN_3O_3)对热带爪蟾胚胎的发育毒性.将3种化合物注入S1一S2期的胚胎后,胚胎的存活率显著下降,其中5 ng TPT,80 ng Rosi和10 ng T0070907注射组存活率分别为46.9%、42.7%和54.2%.胚胎的体长也受到不同程度的影响,5 ng TPT,80 ng Rosi和20 ng T0070907注射组与对照相比体长分别减少了27%、22%和57%.3种化合物还引起了多样的畸形效应,尤其是头部变小及眼睛畸形.说明PPARγ在热带爪蟾早期胚胎的发育特别是头、眼发育中扮演非常重要的角色.TPT与相近剂量T0070907引起的畸形非常相似,说明TPT的致畸机制可能与PPARγ存在某种关系.另一方面,利用整胚原位杂交检测了注射TPT后S20及S25期胚胎的头、眼部标志基因的空间表达,结果表明bf1、en2、krox20及pax6的表达信号均随TPT剂量增大而逐渐变弱且区域变小,定量PCR进一步验证了TPT能在神经胚及早期尾牙期之前影响胚胎头、眼标志基因的表达.研究结果表明,TPT的内暴露对脊椎动物胚胎具有较强的致畸效应和神经毒性.     

三丁基锡抑制雌性斑马鱼卵子发生

【目的】研究三丁基锡(Tributyltin,TBT)长期暴露对雌性斑马鱼(Daniorerio)卵子发生的影响。【方法】用100,500ng·L-1的TBT连续暴露斑马鱼雌性成鱼28d,设置无 TBT 暴露的对照组,并用组织学的方法检测了 TBT 对斑马鱼卵子发育和卵子发生的影响;同时采用定量 RT-PCR 方法研究了 TBT 影响斑马鱼卵子发生的分子机制。【结果】与对照组的情况相比:1)TBT 降低了斑马鱼卵巢质量和性腺成熟系数(GSI),导致早期卵母细胞增多、完全生长期卵母细胞减少和卵母细胞凋亡;2)TBT 上调了20β-hsd,cyp17a2,fshr 和lhr 基因的表达量;下调了孕激素膜受体合成相关基因 mprα 和雌激素合成主要基因cyp19a1a 的表达量。【结论】TBT 可能通过抑制雌激素合成分泌,造成卵子发育受阻;并可能通过抑制孕激素膜受体合成和降低孕激素效应,导致排卵受阻和成熟卵母细胞凋亡。
     

三丁基锡对翡翠贻贝谷胱甘肽硫转移酶活性的影响

在实验条件下,将翡翠贻贝分别暴露于1,10,100,1 000 ng/L的三丁基锡溶液中,1,4,7,14,21 d后取样,测定鳃和消化腺谷胱甘肽硫转移酶(Glutathione S-transferase,GST)同工酶的活性(GSTs,GST-μ,GST-π).结果表明:(1)生理条件下GST同工酶在鳃组织中分布高于消化腺,且活性大小为:GSTs >GST-π>GST-μ;(2)污染条件下,各GST同工酶活性随暴露时间的变化趋势各有不同.鳃和消化腺GSTs活性在暴露第1天被强烈诱导,而后逐步表现为抑制作用;鳃GST-π活性基本无显著变化,消化腺GST-π活性于第7天被诱导至高点,并逐渐恢复到对照水平或被抑制;消化腺GST-μ活性随着污染时间的延长整体受到抑制.     

三丁基锡对热带爪蟾蝌蚪甲调基因mRNA表达的影响

用三丁基锡( TBT)对热带爪蟾(Xenopus tropicalis)NF51阶段蝌蚪进行48 h和96 h暴露.结果显示,50 ng·L-1和200 ng·L-1 TBTCl暴露对胚胎的存活率没有影响.50 ng·L-1TBTCl暴露组TRβ和DI 3 mRNA在蝌蚪尾巴中的表达比对照组增加了1～2.3倍.200 ...     

Spectrofluorimetric determination of artemisinin with pyronine B as the substrate for horseradish peroxidase

Enzyme-catalytic fluorescence determination of artemisinin （qinghaosu, QHS） was developed using pyronine B （PB） as substrate of horseradish peroxidase （HRP）. The interaction between HRP and QHS was an enzyme-substrate model. The catalytic characteristic of HRP in the oxidation reaction, in which the fluorescence of PB was decreased in the presence of QHS, was studied. The steady-state catalytic rate depended upon enzyme and substrate concentrations, and the Michaelis-Menten parameters Km, Vmax and Kcat were 8.4×10^-5mol·L^-1, 7.4×10^-6mol·L^-1s^-1 and 50.23s^-1. The catalytic activity of enzyme was inhibited in the presence of deactivated agents and at high temperature, respectively. Under optimum conditions, linear relationship between fluorescence intensity change （F0-F） of pyronine B and concen- tration of QHS was in the range of 1.41×10^-7-1.27×10^-6mol·L^-1. The detection limit （3σ） was determined to be 2.7×10^-8mol·L^-1. The proposed method was applied to the concentration determination of QHS in the media of plasma or urine samples.     

谷氨酰胺转胺酶发酵培养基的响应面分析优化

利用SAS（Statistical Analysis System）软件中二水平设计的Plackett-Burman设计和响应面分析法（Response Surface Analysis，简称RSA），研究了链霉菌（Streptomyces sp.）HS-1摇瓶发酵生产谷氨酰胺转胺酶的发酵培养基.通过二水平设计实验考察了八种因素对发酵生产谷氨酰胺转胺酶的影响，利用极差分析确定其中以豆饼粉水解物、甘油以及硫酸铵为影响链霉菌发酵生产谷氨酰胺转胺酶的重要因素.通过响应面分析法建立了这三个重要因素的二次回归模型，应用规范分析找到最优点，分别为豆饼粉水解物24.0 g &;#8226; L^-1，甘油24.4 g &;#8226; L^-1，硫酸铵5.40g &;#8226; L^-1.摇瓶实验表明，应用优化培养基谷氨酰胺转胺酶酶活由6.27 μ &;#8226; mL^-1提高到7.30μ &;#8226;mL^-1，提高了16.4%.     

[Zn(bim)2]·(H2O)1.67: A metal-organic open-framework with sodalite topology

Considerable efforts have been focused on the crystal engineering of the supramolecular architectures with desired properties such as zeolitic, magnetic and electric conducting materials[1—3]. But crystal engineering is still a great challenge to the chemists since lack of control over the coordination modes of metal centers and ligands during assembly of extended structures[4]. Some successful synthetic strategies based on topologic control of building blocks have been developed especially i…     

Induction of nicotine in tobacco by herbivory and its relation to glucose oxidase activity in the labial gland of three noctuid caterpillars

Tobacco Nicotiana tabacum L. is a host plant ofHelicoverpa armigera (Huibner), Helicoverpa assulta Guenee and Spodoptera litura (Fabricius) (Lepidoptera, Noctuidae). The difference in leaf nicotine response to the feeding by these three larvae and the mechanical simulation of their feeding was examined by HPLC. Results indicated that nicotine induction was suppressed by H. armigera and H. assulta larvae feeding or by simulated damage treated with their labial glands extracts. The production of nicotine was also suppressed by the glucose oxidase from Aspergillus niger when it was treated on mechanically wounded leaf area. On the contrary, the nicotine production was stimulated by S. litura larva feeding or by simulated damage treated with its labial gland extract. Heat denature can not counteract the stimulation effect of the S. litura labial gland extracts to tobacco nicotine production. The glucose oxidase activity was detected in labial gland extracts of both H. armigera and H. assulta, but the activity in H. armigera was significantly higher than that in H. assulta. No glucose oxidase activity was detected in labial gland extracts of S. litura. It is shown that the glucose oxidase activity in labial glands of caterpillars plays an important role in the nicotine response to herbivory. The glucose oxidase was mainly contained in the labial gland of H. armigera larva, and had the highest activity at pH 7.0. D-Glucose was the optimal substrate of the glucose oxidase. Labial gland glucose oxidase activities varied daily during larval development with high activities found when larvae were actively feeding.     

Mitochondrial response and calcium ion change in apoptotic insect cells induced by SfaMNPV

Mitochondrial responses and changes of calcium ions in apoptotic insect SL-1 cells induced by Syngrapha falcifera multiple nuclear polyhedrosis virus (SfaMNPV) are reported in this paper. By using Rhodamine 123 as a fluorescent labeling probe, flow cytometry analysis and confocal laser scanning microscope observation we observed that the mitochondrial transmembrane potential (△ψm) began to decrease in SL-1 cells at 4 h post infection and △ψm reduced continuously with the extension of virus infection. Western blotting indicated that the Bcl-2 level in the mitochondria gradually declined and was down- regulated. Cells undergoing apoptosis were found to have an elevation of cytochrome c in the cytosol and a corresponding decrease in the mitochondria, which indicated that cytochrome c was released from mitochondria into cytosol. These results suggest that mitochondrion-mediated apoptotic signal transduction pathway exists in apoptotic insect cell induced by SfaMNPV. Cytosolic free calcium ([Ca^2 ]i) concentration rapidly increased after SfaMNPV infection and the elevated calcium was tested to come partly from extracelllular calcium ion influx. Flow cytometry analysis indicated that the apoptosis in SL-1 cells was not influenced by established cytosolic calcium clamped conditions and the EGTA inhibiting calcium influx. Therefore, neither the elevation of cytosolic calcium ion nor extracellular calcium entry was the inducing factor of apoptosis, which hinted that the depletion of ER Ca^2 store contributed to SL-1 cell apoptosis induced by SfaMNPV.     

农田氮素非点源污染模型及年负荷估算研究

研究了农田中氮素随径流动力输出的过程，并依据径流特征和农田氮肥施用情况，将全年区分为水田施肥期、水田生长期和非水田期.按试验时的天气条件，分别采用人工模拟降雨或农田实测降雨的资料，分析农田非点源污染回归模型.水田施肥期采用人工模拟集中降雨情形，降雨强度为2 mm&;#8226;min^-1，模型氮素浓度范围为28~45 mg&;#8226;L^-1；水田生长期和非水田期采用天然降雨，降雨强度为0.037 6~0.075 1 mm&;#8226;min^-1，模型氮素浓度为0.2~4.0 mg&;#8226;L^-1.在分析长系列降雨资料的基础上，综合运用修正SCS（Soil Conservation Service）法、单位线法和相关分析法，估算得到农田径流过程排放的氮素量，再加上地下水渗漏流失的氮素量，取得上海地区一般降水年份农田的氮素非点源污染年负荷量为26.5 kg&;#8226;ha^-1.