间苯二酚与邻苯二酚对泥鳅血清卵黄蛋白原的诱导作用

为探讨间苯二酚(m-dihydroxybenzene)与邻苯二酚(o-dihydroxybenzene)对雄性泥鳅Misgurnus anguillicau-datus血清卵黄蛋白原(Vitellogenin,Vtg)的诱导作用,将雄性泥鳅分别暴露于4种不同质量浓度间苯二酚(5.287mg/L、6.531mg/L、8.072mg/L、10.000mg/L)和邻苯二酚(16.708mg/L、17.742mg/L、18.836mg/L、20.00mg/L)中持续7d、14d、21d和28d,采用碱不稳定性蛋白结合磷法检测血清Vtg水平的变化。结果显示,与对照组相比,泥鳅在低质量浓度间苯二酚(5.287mg/L)和邻苯二酚(16.708mg/L)中暴露7d时,血清Vtg水平均有极显著升高,并且随着暴露浓度的增加,Vtg水平逐渐升高;在同一暴露浓度下,Vtg水平随暴露时间的延长逐渐升高;2种酚类化合物相比,暴露在间苯二酚中的泥鳅血清Vtg水平高于邻苯二酚。上述结果说明,2种化合物均可诱导雄性泥鳅肝脏合成Vtg,具有一定的环境雌激素效应,其诱导作用随着暴露剂量的增大和时间的延长逐渐增强。     

镉胁迫对大弹涂鱼肝脏黄螵呤氧化酶和抗氧化酶活性的影响

研究了不同浓度镉离子对大弹涂鱼肝脏黄嘌呤氧化酶（XOD）、抗氧化酶（超氧化物岐化酶SOD、过氧化氢酶CAT）活性和丙二醛（MDA）含量的影响,以探讨其用于污染暴露的生物标记的可行性.结果表明,低浓度Cd²⁺（0.05 mg·L^-1）暴露使大弹涂鱼肝脏XOD和SOD活性随时间延长升高,第10天达到最大值,中高浓度暴露（0.5 和5 mg·L^-1 Cd²⁺）XOD和SOD活性显著或极显著升高;低和高浓度镉胁迫处理的CAT活性在12 h显著降低,随时间的延长低浓度组CAT活性恢复正常,高浓度组在第7天降到最低值, 并在恢复期的5 d中高浓度组CAT活性却极显著升高;低和中浓度镉胁迫处理的MDA含量12 h极显著升高,而高浓度却极显著下降,随时间延长低浓度恢复正常, 中浓度先上升后下降并到第5天达到最大值,而中高浓度在恢复5 d后MDA含量都极显著降低.     

铜对梨形环棱螺抗氧化酶活性和金属硫蛋白含量的影响

本实验采用暴露重金属的方法,研究了不同浓度硫酸铜（Cu2+ 分别为0、0.005、0.01、0.02、0.05 mg/L）在不同暴露时间（0—14d）下对梨形环棱螺（Bellamya purificata）过氧化氢酶(CAT)、超氧化物歧化酶（SOD）、谷胱甘肽硫转移酶（GST）的活性、还原性谷胱甘肽（GSH）和金属硫蛋白（MT）含量的影响,以探讨Cu2+ 对梨形环棱螺的氧化损伤及其防御作用的机理,并为水环境Cu2+ 污染的早期诊断及生态风险评价提供科学的依据。结果表明：Cu2+对梨形环棱螺肝脏和鳃中CAT、SOD、GST、GSH 和MT 均有明显影响,表现出时间剂量效应。SOD在前4天、CAT在前3天酶活性总体上表现出诱导趋势, GST在前4天酶活性处于诱导状态,随着暴露时间的延长,酶活性下降,到第5天时表现出抑制趋势;随着时间的进一步增长,至14d时, 0.005 mg/L剂量组酶活性维持在正常值附近波动, 0.01 mg/L剂量组酶活性被诱导, 0.02 mg/L剂量组酶活性在肝脏中表现为诱导而在鳃中则被抑制,0.05 mg/L剂量组酶活性被抑制。肝脏和鳃GSH含量的变化与GST相似,在短时间内表现出诱导效应,肝脏GSH在暴露的前5天、鳃GSH在暴露的前4天均处于诱导状态,随着暴露时间的延长,0.005 mg/L剂量组表现出诱导,0.05 mg/L剂量组则受到抑制。MT在整个实验期间均处于诱导状态,各剂量组在0.5d被极显著诱导,随后MT含量出现起伏波动,有上升和下降,至第14天时达到一稳定水平。其中,0.01 mg/L剂量组肝脏的MT在整个实验期间均被极显著地诱导（P <0.01）,0.01 mg/L 剂量组的鳃组织MT除第10天外也被极显著诱导（P <0.01）。在暴露14d时,除0.05 mg/L剂量组的肝脏MT外,其余处于极显著诱导状态（P <0.01）。     

邻苯二甲酸二丁酯(DBP)胁迫对三疣梭子蟹亚急性毒性

研究不同浓度邻苯二甲酸二丁酯(DBP)对三疣梭子蟹相关酶活的影响及组织损伤。设置了三个处理浓度(0.03 mg/L、0.2 mg/L和0.4 mg/L)分别在第0、5、10、15、20 d进行取样。结果表明,随着DBP浓度的增大,三疣梭子蟹死亡率升高。在0.03 mg/L浓度组胁迫下,血清超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和碱性磷酸酶(AKP)活性呈现升高-下降交替波动,过氧化物酶(POD)活性在第10 d低于对照组(P<0.05),其余时间与对照组持平,酸性磷酸酶(ACP)活性在第5 d较对照组显著升高(P<0.05),此后一直保持对照组水平;在0.2 mg/L浓度组胁迫下,血清SOD活性呈现先升高后下降的趋势,POD活性变化不明显,CAT活性先下降后升高,此后恢复到对照组水平,ACP活性在第15 d出现最高值,其余时间与对照组持平,AKP活性呈现先升高后恢复的波动趋势;0.4 mg/L浓度组SOD、CAT和AKP活性呈现先升高后下降的趋势,POD活性和ACP活性在第20 d较对照组低(P<0.05),其余时间与对照组持平。组织切片观察可见,DBP胁迫损伤了三疣梭子蟹肝胰腺和鳃组织,DBP浓度越大,胁迫时间越长,损伤越明显。     

Cd2+胁迫对石菖蒲生理生化特性的影响

通过水堵实验,研究了不同浓度(0、50、200、400μmol/L)下和不同时间内(3、7、11d)Cd2+对石菖蒲(Acorus ta-tarinowii Sehott)叶片抗氧酶活性、叶绿素含量、脂质过氧化程度以及脯氨酸含量的影响.结果表明:(1)超氧化物歧化酶(SOD)活性在3d时随着胁迫浓度的增大而增加,但7d和11d时都是在50 μmol/L浓度下达到峰值,然后降低,但都高于对照;除400μmol/L处理下SOD随胁迫时间延长呈先降后升的趋势外,其余浓度处理下均升高.(2)过氧化物酶(POD)活性在胁迫3d和7d时均随着胁迫浓度的增大而增加,11d时在200 μmol/L处理下达到峰值,然后降低;各浓度处理组均随着胁迫时间的延长POD活性呈下降趋势.(3)过氧化氢酶(CAT)活性在不同时间内随着胁迫浓度的增加呈先增后降趋势,并且各浓度处理组的CAT均随胁迫时间的延长而增加.(4)叶绿素含量在不同胁迫时间内均在200μmol/L处理下达到峰值,然后降低;除400μmol/L处理下的叶绿素随胁迫时间延长而增加外,其余浓度处理组均呈下降趋势.(5)丙二醛(MDA)含量和脯氨酸(Pro)含量均随着胁迫浓度的增加和胁迫时间的延长而增加.研究结果分析表明石菖蒲对Cd2+胁迫有一定的耐受性.     

镉对尖紫蛤抗氧化酶活性及脂质过氧化的影响

为阐明镉(Cd2+)对尖紫蛤消化盲囊和鳃抗氧化酶的毒性影响程度,研究了不同浓度的Cd2+(0.005、0.05、0.5 mg/L)在不同暴露时间(24h、72h、120h)对尖紫蛤鳃和消化盲囊中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)的活性以及丙二醛(MDA)含量的影响。结果表明,在Cd2+浓度为0.005 mg/L时,在整个实验期间Cd2+对消化盲囊和鳃内的SOD、CAT、GSH-PX活性并无显著影响。在Cd2+浓度为0.05 mg/L和0.5 mg/L时,SOD、CAT、GSH-PX在鳃和消化盲囊中的活性都呈现出明显的时间剂量依赖关系。在0.05 mg/L暴露时,鳃和消化盲囊中的SOD、CAT和GSH-PX的活性随时间逐渐增强,在72h时达到最大值,但在120h时略有降低。在0.5 mg/L暴露时,消化盲囊中SOD、CAT及鳃中CAT活性在24h时上升达到最大值,但鳃中SOD直到72h才达到最大值,并均在120h下降到最低,其中消化盲囊中SOD和CAT活性在120h低于对照组,这可能与消化盲囊对Cd2+的敏感性高于鳃有关。在0.5 mg/L暴露的鳃中,GSH-PX在24h、72h活性并不上升,在120h甚至低于正常水平,0.5 mg/L暴露的消化盲囊中,24h时迅速增高,然后逐渐下降到正常值。这可能与Cd2+结合了GSH-PX的活性中心,降低了GSH-PX的活性有关。与3种酶活性随着时间延长和剂量的增加,酶活性会降低的变化趋势不同,鳃和消化盲囊中的MDA的含量随时间延长和剂量的增加而增加,并不出现下降,这表明尖紫蛤鳃和消化盲囊中的MDA含量可以灵敏的反映机体内的氧化损伤程度,但不能敏感的反映水体中Cd2+的污染情况。     

重金属Cd胁迫对红树蚬的抗氧化酶、消化酶活性和MDA含量的影响

摘要：本文在实验室条件下,观测不同Cd胁迫处理（时间和水平）对红树蚬（Geloina coaxans (Gmelin)）的胃组织中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、丙二醛(MDA)含量,及淀粉酶、脂肪酶和蛋白酶等3种消化酶活性的影响效应。结果表明,高浓度组（4.00 mg?L-1和8.00 mg?L-1）在较短的暴露时间即出现SOD 活性显著诱导,而低浓度组则需要更长的暴露时间。所有处理组的CAT活性均先受诱导而后抑制,但去除胁迫后低浓度组活性上升,高浓度组则持续下降。低浓度处理组在胁迫初期MDA含量上升,但随后下降至较低水平;高浓度处理组MDA含量上升稍为滞后,但随后上升至较高水平。在消化酶方面,Cd对红树蚬胃组织淀粉酶的影响基本上表现为抑制;在胁迫初期脂肪酶活性受到显著诱导,随着胁迫时间延长酶活性则下降,同时胁迫解除后不同程度恢复;对蛋白酶的影响效应规律性不明显。显著的效应-剂量间相关关系存在于特定时间的SOD活性（1d）,CAT、淀粉酶活性和MDA含量（7d和恢复6d）。本文还探讨了这些指标作为生物标记物应用于监测海洋重金属污染的可能性。 关键词：红树林;红树蚬;Cd胁迫;消化酶;抗氧化酶;脂质过氧化     

铜胁迫对穿心莲幼苗生长及生理特性的影响

通过对盆栽穿心莲(Andrographis paniculata)幼苗浇灌Cu SO4溶液实验,研究外施Cu2+对穿心莲幼苗的生理特性和药效成分含量的影响。结果显示,外施Cu2+对穿心莲幼苗的生长、生理生化指标和穿心莲内酯、脱水穿心莲内酯的含量均有显著影响。当Cu SO4溶液浓度超过6.25 mmol/L时,Cu2+能显著抑制穿心莲幼苗株高、叶长、叶宽的增长,且随胁迫时间的延长和Cu2+浓度的增大抑制作用增强;穿心莲叶片中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量均随Cu2+浓度增大和胁迫时间的延长而升高,当Cu SO4溶液浓度达到12.5 mmol/L且胁迫20 d和30 d时,叶片中SOD活性达到对照组的168.3%和171.18%;过氧化物酶(POD)、过氧化氢酶(CAT)活性则随Cu2+浓度的增大呈先升高后下降的趋势;当外施Cu SO4溶液浓度大于1.25 mmol/L时,与对照相比,穿心莲药效成分穿心莲内酯和脱水穿心莲内酯的含量均显著降低(P0.05)。研究结果表明,外施Cu SO4溶液浓度高于6.25 mmol/L时,能显著影响穿心莲幼苗的生理特性和有效成分含量,从而降低穿心莲药材的产量和质量。     

镉长期暴露对黑斑蛙的氧化胁迫和抗氧化能力的影响

在实验条件下,将黑斑蛙暴露于12．5mg／L和25．0mg／L浓度的镉溶液中30d,分别测定了黑斑蛙在暴露10、20和30d时肝、肾组织中镉（Cd）含量、过氧化产物丙二醛（MDA）的含量、还原型谷胱甘肽（GSH）含量和超氧化物歧化酶（SOD）活性,以探讨镉对机体的脂质过氧化作用及机体的抗氧化损伤机制。实验结果表明,不同剂量组黑斑蛙肝、肾镉含量、MDA含量均随着镉暴露时间的延长而升高,且肝MDA含量与镉在肝中的蓄积量呈显著正相关（R^2=0．8643,n=9）。肝脏GSH含量随镉暴露时间的延长而被显著诱导,且与MDA含量呈显著正相关（R^2=0．5933,n=9）;肾GSH含量则随暴露时间的延长而显著下降,与MDA含量呈显著负相关（R^2=0．8609,n=9）。不同剂量组肝SOD活性随镉暴露时间的延长而升高,肾SOD活性在高剂量组随镉暴露时间的延长表现为先升高后回落下降的趋势。可见,在镉的长期暴露下,细胞膜过氧化增强是镉伤害机体的主要原因,而GSH含量、SOD活性的升高则可能是机体抗过氧化的机理之一。     

重金属Cu2+、Pb2+单因子及联合毒性对泥鳅卵细胞DNA的损伤效应

采用室内暴露试验方法, 研究了不同浓度Cu2+(0.01、0.10、0.25mg/L)、Pb2+(0.05、0.50、0.75mg/L)单因子染毒以及Cu2++Pb2+(0.01 mg/L+0.05mg/L、0.10 mg/L+0.50 mg/L、0.25 mg/L+0.75 mg/L)联合染毒对泥鳅卵细胞DNA的损伤效应, 并以SCGE技术进行检测。结果显示, Cu2+与Pb2+单因子染毒对泥鳅卵细胞DNA的损伤具有较为显著的剂量-效应与时间-效应关系(P0.05)。Cu2++Pb2+联合染毒, 在溶液暴露的0-5d表现为剂量-效应与时间-效应关系(P0.05); Cu2++Pb2+暴露5-10d 则表现出拮抗作用。研究结果显示, Cu2+、Pb2+ 单因子及联合染毒均造成泥鳅卵细胞DNA损伤, 具有基因毒性效应。       

Molecular Characterization of Japanese Sillago Vitellogenin and Changes in Its Expression Levels on Exposure to 17β-Estradiol and 4-<Emphasis Type="Italic">tert</Emphasis>-Octylphenol

We cloned a full-length cDNA encoding vitellogenin (VTG) from a marine teleost, the Japanese sillago Sillago japonica. The cloned sillago VTG contained signal peptide, lipovitellin heavy chain, phosvitin, lipovitellin light chain, and β′-component in the order from the N-terminus. An exposure to 17β-estradiol significantly increased the levels of plasma VTG, but not hepatic VTG mRNA in males. Neither plasma VTG nor hepatic VTG mRNA levels were affected by the exposure to 4-tert-octylphenol. Hepatic VTG mRNA levels in males increased at 1 day after intraperitoneal administration of 17β-estradiol but decreased in the subsequent 5 days. However, plasma VTG levels remained high for 5 days after administration, suggesting that the accumulation period of plasma VTG is longer than that of hepatic VTG mRNA in males. Therefore, VTG mRNA may be a suitable indicator of temporal exposure to estrogenic chemicals in the environment, whereas plasma VTG is useful to detect consecutive exposure.     

Effects of heavy metals Cd2+, Pb2+ and Zn2+ on DNA damage of loach Misgurnus anguillicaudatus

The effects of heavy metals Cd²⁺, Pb²⁺ and Zn²⁺ at 0.05, 0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1–35 days exposure were examined by single cell gel electrophoresis (SCGE). For each test group, 20 loaches with similar body size (5.17–7.99 g; 11.79–13.21 cm) were selected and kept in aquaria with dechlorinated water at (22±1)°C and fed a commercial diet every 48 h. According to the percentage of damaged DNA with tail and its TL/D (tail length to diameter of nucleus) value, the relationship between DNA damage degree and heavy metal dose and exposure time was determined. Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time. The highest percentage (84.85%) of damaged DNA was shown in 5.0 mg/L Zn²⁺ group after 28 days exposure and the biggest TL/D value (2.50) in all treated groups after 35 days exposure. During the first treated week, the damnification of DNA was mainly recognized as the first level, after that time, the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%. The joint toxic effects among Cd²⁺, Pb²⁺ or Zn²⁺ revealed much complexity, but it generally displayed that the presence of Cd²⁺ could enhance the genotoxicity of Pb²⁺ or Zn²⁺. In conclusion, the results suggested that there was a significant time-and dose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach, and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms. __________ Translated from Acta Hydrobiologica Sinica, 2006, 30(4): 399–403 [译自: 水生生物学报]     

Endocrine disrupters with (anti)estrogenic and (anti)androgenic modes of action affecting reproductive biology of Xenopus laevis: I. Effects on sex steroid levels and biomarker expression

Adult Xenopus laevis were exposed in vivo to ethinylestradiol, tamoxifen, methyldihydrotestosterone and flutamide as (anti)estrogenic and (anti)androgenic compounds, respectively, for four weeks at a concentration of 10(-8) M and to Lambro river water, a polluted river from Italy. Effects of the treatments were analysed by mRNA expression of retinol-binding protein (RBP), transferrin (TF), transthyretin (TTR) and vitellogenin (VTG) in the liver of male and female X. laevis, to analyse the potential of these genes to detect endocrine disrupting compounds (EDC) with different modes of action. In addition, plasma VTG and sex steroid levels, estradiol-17beta (E(2)) and testosterone (T), were analysed. Sex steroids were depressed by ethinylestradiol in both sexes whereas tamoxifen increased E(2) in females. The induction of VTG protein plasma levels was more pronounced at the protein level compared to hepatic VTG mRNA expression in response to estrogenic treatment but VTG mRNA expression detected both, estrogenic and antiestrogenic EDC. The mRNA expression of TF was decreased by estrogenic and increased by antiestrogenic treatment while TTR mRNA expression was down-regulated and RBP mRNA up-regulated by estrogenic exposure. The other treatments did not affect the mRNA expression of the examined genes.     

Genotoxicity and Estrogenic Activity of 3,3′-Dinitrobisphenol A in Goldfish

3,3′-Dinitrobisphenol A (dinitro-BPA) is formed in a mixture of bisphenol A (BPA) and nitrite under acidic conditions. It shows genotoxicity in male ICR mice on a micronucleus test, but its estrogenic activity has not been examined in vivo. We examined its estrogenic activity using goldfish (Carassius auratus) by measuring plasma levels of vitellogenin (VTG) by the ELISA method. Expression of VTG didn’t increase in the plasma of goldfish intraperitoneal injected with dinitro-BPA at a dose of 10 mg/kg of body weight.

We also examined the genotoxicity of dinitro-BPA by single-cell gel electrophoresis (comet assay) and a micronucleus test using goldfish. The DNA tail moment of blood cells increased after intraperitoneal injection of dinitro-BPA. Dinitro-BPA at the same dose significantly increased micronucleus frequency in gills of goldfish. On the other hand, BPA did not significantly increase the frequency of micronucleated cells.

In conclusion, we found that dinitro-BPA did not show estrogenic activity, but had genotoxic potency stronger than that of BPA.     

一株耐盐日本曲霉的筛选及其溶磷促生作用

【目的】从内蒙古种植葵花的盐碱地中筛选高效溶磷真菌,为盐碱地增产节肥开发生物肥料提供溶磷菌种资源。【方法】利用ITS r DNA序列鉴定菌株、固体培养基测定耐盐性,液体摇床培养与盆栽试验结合分析菌株溶磷能力,盆栽和田间试验明确菌株M1促进作物生长和增产作用;LC-MS技术测定菌株M1在液体培养基中分泌有机酸和植物激素含量,明确菌株M1的溶磷和促生机理。【结果】溶磷菌株M1鉴定为日本曲霉(Aspergillus japonicus)。液体培养基接种菌株M1培养6 d,以Ca_3(PO_4)_2为磷源时上清液有效磷达1020.89 mg/L,溶解率为63.30%;以AlPO_4为磷源时有效磷达995.69 mg/L,溶解率为48.59%;以贵州开阳磷矿粉、江苏锦屏磷矿粉、云南晋宁磷矿粉、河北钒山磷矿粉和云南昆阳磷矿粉为磷源接种菌株M1,从晋宁磷矿粉释放的有效磷浓度最高,达到363.64 mg/L。菌株M1可耐受10%NaCl。将M1制备的菌剂分别接种于施用Ca_3(PO_4)_2、AlPO_4和开阳磷矿粉3种磷源的4种盆栽试验土壤包括北京石灰性潮土、安徽黏性潮土、安徽水稻土和山东沿海盐潮土。结果显示,菌株M1对玉米植株促生效果显著,玉米植株鲜重比对照提高2.14%–90.91%、干重增加22.15%–268.28%;土壤有效磷提高21.81–24.27 mg/kg。菌株M1与4种土壤的适配性均高于对照菌株DSM 821。田间小区花生产量结果显示,接种溶磷菌剂M1增产效果最好,花生果实产量达4.46 t/hm~2,比不接种菌剂的对照处理增加0.81 t/hm~2,增产22.19%。菌株M1在含有磷酸三钙、磷酸铝和开阳磷矿粉3种难溶磷培养液中经过6 d培养,均产生7种有机酸,其中草酸和柠檬酸含量最高,分别为616.16 mg/L和413.69 mg/L;培养液均能检测到吲哚乙酸(IAA)和玉米素,IAA含量为15.45–77.58 mg/L,玉米素浓度为0.06–0.11 mg/L。【结论】获得了一株高效溶解多种难溶磷的日本曲霉菌M1,它能显著增加土壤有效磷、促进玉米生长和花生增产,与4种典型土壤适配性好,具有良好的农业应用前景。     

Antioxidative response of Lemna minor plants exposed to thallium(I)-acetate

The physiological effects of thallium(I)-acetate on the duckweed Lemna minor after 1-, 4-, 7- and 14-d exposure were analyzed. High bioaccumulation of Tl (221 mg kg⁻¹ dry wt at 2.0 μM Tl-acetate) caused an inhibition of plant growth. After 14-d exposure, 0.2, 0.5, 1.0 and 2.0 μM Tl-acetate reduced the frond-number growth rate by 21.1%, 39.4%, 66% and 83.1%, respectively. Tl-acetate also induced a modulation of the antioxidative response by depleting the ascorbate content and affecting the antioxidative enzymes activities. Superoxide dismutase showed a continuous increase of activity (31–67%) after Tl-acetate exposure. Other antioxidative enzymes displayed a biphasic response to both the concentration and the exposure period. Exposure up to 7 d decreased the catalase activity (up to 40%) in plants treated with higher Tl-acetate concentrations. In contrast, 14-d exposure increased the activity of the enzyme (≥90%). Short-term exposure increased ascorbate peroxidase activity (13–41%), except in plants exposed to the highest Tl-acetate concentration. However, 14-d exposure decreased the enzyme activity at all concentrations tested (38–60%). Although pyrogallol peroxidase activity increased (up to 26%) during 4-d exposure, longer exposures to the highest two concentrations decreased the activity of the enzyme (25–48%). In general, short-term exposure to Tl-acetate activated the antioxidant capacity, which resulted in recovery of the frond-number growth rates in Tl-treated plants. In spite of the activation of the antioxidative response during short-term exposure, higher Tl-acetate concentrations increased the hydrogen peroxide level (up to 45%) and induced marked oxidative damage to lipids, proteins and DNA. Longer exposure induced a decline of the antioxidative response, and plants showed the symptoms of oxidative damage even at lower Tl-acetate concentrations. The genotoxic effect was evaluated by an alkaline version of the cellular and acellular Comet assay, which revealed an indirect genotoxic effect of Tl-acetate, suggesting oxidatively induced damage to DNA.     

Effect of triethylenepentaminehexaacetic acid on the renal damage in cadmium-treated Syrian hamsters

Cadmium (Cd)-induced nephropathy was treated by triethylene-pentaminehexaacetic acid (TTHA) in male Syrian hamsters. Hamsters injected three times a week with 3 mg/kg body wt CdCl₂ showed proteinuria, urinaryN-acetyl-β-d-inglucosaminidase (NAG), and fractional excretion of sodium (FENa) when compared to saline-injected control. Cd-treated hamsters injected ip with TTHA 10 mg/kg body wt five times a week showed reduction of renal damage, including reductions in urinary protein (from 6.7±2.2 to 4.3±0.5 mg/d) and NAG (0.17±0.06 to 0.04±0.02 U/d). Urinary excretion of Cd was significantly increased (from 87±51.3 to 3052±1485 mg/L) by TTHA administration. Cd concentration in renal cortical tissue was slightly reduced (26.4±3.0 to 21.8±2.7 mg/g. protein). Excretion of malondialdehyde (MDA) was increased only in Cd-injected hamsters (to 2.1±1.6 nM/L), and elevated MDA in renal cortical tissue was not reduced by the administration of TTHA (1041±105 vs 1104±358 nM/g protein). Glutathione (GSH) concentration in the renal cortex was significantly elevated after Cd administration and further increased after TTHA administration (5.5±2.1 to 9.8±2.0 μg/50 mg protein). There were no marked effects on creatinine clearance (Ccr) and hematocrit. Moreover, renal morphological changes were improved significantly by treatment with TTHA. We demonstrated the efficacy of TTHA in the treatment of Cd-induced nephropathy in hamsters. Although the precise mechanism of the TTHA effects on Cd-induced nephropathy has not been elucidated, it might involve GSH reducing the elevated MDA concentration in renal tissue.     

双酚A诱导雄性中国林蛙肝细胞雌激素受体表达和卵黄蛋白原合成

为探讨双酚A(bisphenol A,BPA)对雄性两栖动物肝细胞中雌激素受体(estrogenreceptor,ER)表达和卵黄蛋白原(vitellogenin,VTG)合成的影响,将中国林蛙(Ranachensinensis)雄性成体分别持续暴露于10-7、10-6、10-5mol/LBPA水体中10、20、30d,设10-9、10-8mol/L雌二醇(E2)为阳性对照。用原位杂交技术检测ERmRNA在肝细胞中的表达定位,用免疫组织化学技术检测肝细胞中ER和VTG蛋白的表达。结果显示,各BPA和E2处理组肝细胞中均有ERmRNA阳性反应,ER和VTG蛋白的表达相对值均显著高于空白对照组。在同一暴露时间,ER和VTG表达值是随着BPA浓度的增加呈增高趋势。在同一BPA浓度,VTG表达值是随暴露时间的延长呈增高趋势,而ER表达值则无显著性变化。这些表明BPA可以通过诱导雄性中国林蛙肝细胞的ER高调导致VTG合成,但其效应远低于E2。     

Generating oxidation-resistant variants of <Emphasis Type="Italic">Bacillus kaustophilus</Emphasis> leucine aminopeptidase by substitution of the critical methionine residues with leucine

Bacillus kaustophilus leucine aminopeptidase (bkLAP) was sensitive to oxidative damage by hydrogen peroxide. To improve its oxidative stability, the oxidation-sensitive methionine residues in the enzyme were replaced with leucine by site-directed mutagenesis. The variants, each with an apparent molecular mass of approximately 54 kDa, were overexpressed in recombinant Escherichia coli M15 cells and purified to homogeneity by nickel-chelate chromatography. The specific activity for M282L, M285L, M289L and M321L decreased by more than 43%, while M400L, M426L, M445L, and M485L showed 191, 79, 313, and 103%, respectively, higher activity than the wild-type enzyme. Although the mutations did not cause significant changes in the K _m value, more than 67.8% increase in the value of k _cat/K _m was observed in the M400L, M426L, M445L and M485L. In the presence of 50 mM H₂O₂, most variants were more stable with respect to the wild-type enzyme, indicating that the oxidative stability of the enzyme can be improved by engineering the methionine residues. This revised version was published online in June 2006 with corrections to the Cover Date.