三种氨基甲酸酯类农药化合物对DNA的潜在损伤作用

应用紫外光谱法研究了3种氨基甲酸酯类农药对DNA的损伤作用，结果显示，这3种氨基甲酸酯类农药能够引起ctDNA的紫外吸收光谱发生发射波长以及吸收强度的变化，从而说明3种氨基甲酸酯农药能够与ctDNA进行作用，即意味着3种氨基甲酸酯类农药进入生物有机体后有可能通过形成DNA加合物的形式而进一步导致基因突变，最终对生物体的DNA产生化学损伤。     

稳定同位素稀释GC-MS测定不同年份葡萄酒中的氨基甲酸乙酯

改进了气相色谱—质谱(GC-MS)测定葡萄酒中氨基甲酸乙酯(EC)的方法,检测和分析了不同年份葡萄酒中的EC含量。葡萄酒样品加入EC-d5内标,经硅藻土吸附、二氯甲烷洗脱、减压浓缩、氮气吹干和甲醇复溶,气相色谱—质谱选择离子检测。方法检出限和定量限分别不大于2ng/mL和5ng/mL,验证的浓度范围内(5~100ng/mL),保持良好线性,r2=0.999 8,回收率为89.9%~100.1%,相对标准偏差为2.91%~4.76%。对4个品种、3个年份的葡萄酒中EC的评价结果表明,年份越早葡萄酒中EC含量越高;贮藏时间是导致葡萄酒中EC含量升高的主要因素。     

环境样品氨基甲酸酯类农药的分离和检测技术研究进展

氨基甲酸酯类农药是一类高效广谱型杀虫剂，近年来在农业生产中被广泛使用，然而由此带来的环境污染和食品健康问题也日益引起关注。无论是样品的前处理方法还是仪器检测方法都有待不断地完善和发展，因此，开发理想的样品前处理方法和更加有效的仪器分析方法成为该领域的研究热点。本文在查阅了近年国内外相关文献61篇的基础上，就环境样品氨基甲酸酯类农药的分离提取方法以及不同方法的优缺点做分析，同时对氨基甲酸酯类农药的仪器分析方法及其发展趋势做总结，以期为研究环境样品氨基甲酸酯类农药的分析技术提供有益的参考。     

Acute aldicarb toxicity in dogs: 15 cases (2001–2009)

Objective – To describe the common clinical signs, laboratory abnormalities, treatment, and prognosis associated with acute aldicarb toxicosis in dogs. Design – Retrospective observational study from 2001 to 2009. Setting – Urban referral hospital. Animals – Fifteen client‐owned dogs. Interventions – None. Measurements and Main Results – The most common clinical signs associated with acute aldicarb toxicosis were vomiting, ptyalism, diarrhea, and tremors. Of the 15 dogs, 11 were admitted to the hospital for treatment, 2 were euthanized at presentation and 2 were discharged against medical advice following minimal treatment and lost to follow‐up. Laboratory abnormalities included lactic acidosis and hyperglycemia in 12 and 9 patients, respectively. Treatment of hospitalized dogs included induction of emesis with apomorphine (4 dogs), activated charcoal (5), IV fluids (11), atropine (7), methocarbamol (3), diazepam (1), pralidoxime (1) and diphenhydramine (1). Ten of 11 hospitalized dogs survived to discharge; 1 was euthanized following a respiratory arrest after 36 hours of hospitalization. One patient received mechanical ventilation and treatment for pneumonia before discharge from the hospital. The median duration of hospitalization was 22 hours (range 12–168 h). Conclusions – Acute aldicarb toxicosis carries a good prognosis for survival and hospital discharge with treatment. Supportive care should be considered for at least 18–24 hours to monitor for response to therapy and development of respiratory failure.     

杀螺药物研究──碳酸酯、氨基甲酸酯类化合物的合成及其构效关系初步探讨

合成了碳酸酯类化合物23个和苯氨基甲酸苯酯类化合物7个。杀螺初筛试验发现，8个碳酸酯化合物具有杀螺活性，其中，化合物I1和I11，10ppm的杀螺率为100％和96．7％。碳酸苯酯的苯环上，用硝基或3个以上的氯原子取代，有利于增强该类化合物的杀螺作用。杀螺效果较好的三氯苯碳酸酯改变成同电异素体类似物，苯氨基甲酸酯后，杀螺作用明显下降。     

Carbamate poisoning in a dairy goat herd: Clinicopathological findings and therapeutic approach

Abstract

CASE HISTORY: Approximately 1 hour after the consumption of carnations from a nearby glasshouse 55 animals from a dairy goat herd exhibited signs of possible poisoning.

CLINICAL FINDINGS: Upon clinical examination affected animals exhibited signs of salivation, tympany, tachypnoea, polydipsia, urination, diarrhoea, bradycardia, miosis, tremor and convulsions. As poisoning from an acetylcholinesterase-inhibiting insecticide was suspected, treatment with atropine sulphate was initiated at a dose of 0.3 mg/kg bodyweight. The treatment was repeated for some animals that relapsed, and was effective in all cases, with the exception of one goat kid that died.

DIAGNOSIS: Necropsy of the goat kid showed pulmonary oedema and congestion of internal organs. Toxicological analysis of stomach contents and liver of the dead animal, as well as of the carnations, revealed high concentrations of carbamates.

CLINICAL RELEVANCE: Carbamate poisoning after consumption of polluted feedstuffs or glasshouse products can be fatal for dairy goats. Atropine sulphate, at 0.3 mg/kg, can be useful in treating this condition.     

Effects of tea tree oils on removing pesticide residue in cowpea

为研究茶树油清除果蔬农药残留的效果,该试验选取豇豆为供试材料,以不同浓度的茶树油和水溶性茶树油等清洗处理,利用气相色谱和气相色谱-质谱联用检测豇豆内有机磷类、拟除虫菊酯类和氨基甲酸酯类的农药残留量,计算农药清除率。供试7种农药中,水胺硫磷、马拉硫磷、氧乐果、三唑磷、毒死蜱、氯氰菊酯和速灭威在豇豆中的初始浓度分别为：20.395、1.690、6.524、10.719、0.160、12.104和23.057 mg/kg。茶树油处理后检测结果表明,茶树油具有清除残留在豇豆中农药的能力,清除效果随茶树油浓度增加而增强;清除有机磷类农药效果较拟除虫菊酯类和氨基甲酸酯类农药明显。茶树油比去离子水、市售果蔬农残清洗剂清除农药残留效果显著,同时,相同浓度的水溶性茶树油比相应茶树油清除农药残留能力强。0.8%水溶性茶树油清除效果最佳,清除率分别为水胺硫磷80.48%,马拉硫磷94.54%,三唑磷82.79%,毒死蜱84.58%,氧乐果72.20%,氯氰菊酯80.51%,速灭威72.21%。通过研究结果可知,茶树油可作为有开发前景的果蔬清除剂。     

家蚕乙酰胆碱酯酶基因bmace在毕赤酵母GS115中的表达及其分析

【目的】构建表达家蚕乙酰胆碱酯酶（BmAChE）的毕赤酵母菌株,实现BmAChE在毕赤酵母中分泌表达并对其活性进行分析。【方法】从家蚕头部提取总RNA,经RT-PCR反转录成cDNA,通过PCR扩增出家蚕乙酰胆碱酯酶基因bmace并进行序列测定。采用PCR及重叠延伸PCR去除原bmace中的信号肽和疏水肽,并对序列中存在的2个A+T富含区进行同义密码子替换的改造。将改造后的bmace与表达载体pPIC9K连接,转化毕赤酵母GS115,甲醇诱导表达。SDS-PAGE和Western blotting鉴定重组蛋白,以Ellman法测定酶活力,毒扁豆碱和有机磷及氨基甲酸酯类农药抑制试验分析重组BmAChE（rBmAChE）的生物活性。【结果】改造后的bmace经重组毕赤酵母菌株分泌表达了1个分子量约为70 kD的蛋白,具有AChE活性,酶比活力约为1 187 U•mg-1。抑制试验显示,该rBmAChE能够被毒扁豆碱强烈抑制,并且可被皮蝇磷等10种有机磷农药和克百威等5种氨基甲酸酯类农药抑制,最低抑制浓度IC50值可达2.78×10-10 mol•L-1。【结论】成功构建分泌表达BmAChE的毕赤酵母菌株,其表达了具有良好活性的rBmAChE,可用于进一步制备有机磷及氨基甲酸酯类农药快速检测分析制剂。     

Studies on the Purification and Characterization of Soybean Esterase, and Its Sensitivity to Organophosphate and Carbamate Pesticides

Soybean esterase, a cholinesterase-like enzyme, was purified by differential centrifugation firstly, then, ammonium sulfate precipitation, dialysis, and finally, DEAE-cellulose-32 ion-exchange chromatography after extracting it from soybean seeds with phosphate buffer （0.3 mol L^-1, pH 7.0）. The extract recovery rate of the purified enzyme was 8.18% and purification fold was 91.58. The soybean esterase appeared as two bands on the denaturing SDS-PAGE with molecular weights of 24 and 37.2 kDa, respectively, which proved that it is a dimer protein consisting of two subunits. The result of nondenaturing PAGE revealed that the soybean esterase is a single band with cholinesterase-like activity using α- naphthyl acetate as the substrate and fast blue B salt as coloring agent. The esterase showed very high sensitivity to 18 kinds of organophosphate pesticides and 6 kinds of carbamate pesticides with the lowest detective limits of 0.03125- 0.0625 and 0.03125-0.25 mg kg^-1, respectively, and can meet the demands of MRL specified by the most countries.     

新型固定化乙酰胆碱酯酶传感器的制备及氨基甲酸酯农残检测

以双醛纤维素(dialdehyde cellulose,DAC)为固载酶基质材料,通过固定化酶化学键联技术制备固定化乙酰胆碱酯酶(DAC-AChE),采用自组装法制备多层壳聚糖/固定化乙酰胆碱酯酶/功能化碳纳米管生物传感器(Chi/DAC-AChE/F-CNTs/GCE),利用交流阻抗法(AC impedance,ACI),循环伏安法(cyclic voltammetry,CV)及差分脉冲伏安法(differential pulse voltammetry,DPV)考察新型传感器对农残的检测性能。结果表明:成功制备了新型固定化乙酰胆碱酯酶电化学传感器;新型传感器对氨基甲酸酯杀虫剂1-萘基-N-甲基氨基甲酸酯(carbary,CAB)具有良好的响应,检测方法响应快速,线性范围宽,灵敏度高,最低检出限可达3.5 μg·L-1,具有很好的重现性和稳定性,有关结果可为氨基甲酸酯类杀虫剂的检测提供一种新的分析方法。