纳米铜体内体外急性毒性对比研究

目的分别使用昆明小鼠急性经口毒性试验、秀丽隐杆线虫毒性试验及CHL体外细胞毒性试验研究纳米铜的急性毒性作用。方法小鼠急性经口毒性试验采用霍恩氏法;线虫急性毒性试验对秀丽隐杆线虫L4期幼虫进行24 h染毒;体外细胞毒性试验采用CHL细胞中性红染色法。结果纳米铜小鼠急性经口毒性试验雌性小鼠经口半数致死量LD50为14.0 mg/kg·bw,雄性小鼠经口半数致死量LD50为8.8 mg/kg·bw;线虫急性毒性试验显示,随着纳米铜染毒剂量的增加,线虫死亡率呈上升趋势,纳米铜对线虫IC50为12.6μg/ml;细胞毒性试验显示,随着纳米铜染毒剂量的增加,CHL细胞存活率呈下降趋势,纳米铜对CHL细胞IC50为2.33μg/ml。结论根据化学品急性经口毒性试验方法(GB/T 21603-2008)的分级标准判定,纳米铜小鼠急性经口毒性属高毒;12.9和30μg/ml纳米铜对线虫有毒性作用;大于3.75μg/ml纳米铜细胞毒性为4级。     

硫酸铝钾对小鼠的急性毒性和蓄积毒性试验

目的研究硫酸铝钾[Aluminium potassium sulphate,KAl(SO4)2]对小鼠的急性毒性、蓄积毒性以及神经毒性。方法采用霍恩氏法测定KAl(SO4)2小鼠急性经口毒性,即半数致死量(Median lethal dose,LD50);用定期递增剂量法测定KAl(SO4)2经口染毒蓄积毒性,采用自身对照,并以平面翻正试验,断崖回避试验以及游泳试验测定KAl(SO4)2对小鼠的神经行为毒性。结果 KAl(SO4)2经口染毒对雄、雌小鼠LD50值分别是4 300和3 690 mg/kg;定期递增计量法灌胃28 d后,雄、雌小鼠蓄积系数分别是K=9.97、K12.8;行为毒理学试验中小鼠游泳试验随着染毒剂量和时间增加得分明显减少(P0.05),其他试验结果无明显差异。结论在430~4 816 mg/kg和368~4 132.8 mg/kg剂量范围内,KAl(SO4)2对雄、雌小鼠蓄积毒性分别为轻度蓄积和无蓄积毒性,但对染毒动物的神经行为有一定影响。     

3种急性经口毒性试验方法的比较

传统的急性经口毒性试验多采用霍恩氏法。该法使用动物较多，造成人力和物力的浪费，固定剂量法和毒性等级法是2种主要的LD50测试替代方法。固定剂量法由英国毒理学会（BTS）于1984年正式提出，目前Bmsels委员会也承认其具有一定的实用价值。毒性等级法最初由Samiey提出，现已被许多欧美国家作为公认的LD50测试修正方法之一。此2种方法主要流行于欧洲国家，目前在国内还较少使用。     

20%氰戊菊酯乳油急性经口暴露对雄性大鼠血清生化指标的影响

目的了解20%氰戊菊酯乳油急性经口暴露引起的雄性SD大鼠血清生化指标的改变情况。方法霍恩氏法确定20%氰戊菊酯乳油对雄性大鼠LD50,测定20%氰戊菊酯乳油3个染毒剂量(1/2LD50、1/4LD50和1/8LD50)组动物及溶剂对照组动物在急性经口暴露后0. 5、1和2 h 3个时间点的18项血清生化指标。结果染毒组大鼠总蛋白、白蛋白、尿素氮、肌酐和血糖较对照组均有明显升高(P0. 05);天冬氨酸转氨酶、尿酸、总胆固醇、高密度脂蛋白、蛋白质羰基、超氧化物歧化酶和还原型谷胱甘肽较对照组均有明显降低(P0. 05)。结论 20%氰戊菊酯乳油急性经口暴露引起了雄性大鼠血清生化指标的异常改变。     

转HJC-1和G6-EPSPS基因抗虫耐草甘膦水稻表达蛋白的小鼠急性经口毒性研究

目的探讨转HJC-1和G6-EPSPS基因抗虫耐草甘膦水稻表达蛋白的急性经口毒性。方法选择体重18～22 g、SPF级健康昆明种小鼠,禁食16 h后,G6蛋白以1693.3 mg/kg.BW剂量,HJC-1蛋白以751和1 502 mg/kg.BW2个剂量经口灌胃染毒,连续观察2周。结果 G6蛋白和751 mg/kg.BW剂量的HJC-1蛋白染毒后未见明显中毒症状,观察期内无死亡;1502 mg/kg.BW剂量的HJC-1蛋白雌雄各死亡1只。结论 G6表达蛋白对小鼠急性经口毒性LD50>1693.3 mg/kg.BW;HJC-1表达蛋白对小鼠急性经口毒性LD50>751 mg/kg.BW。     

血管生长抑制因子Kringle 5的发酵条件研究

确定了适合重组工程菌生长和血管生长抑制因子Kringle 5表达的培养基,并采用正交试验优化了重组工程菌的发酵条件.20L发酵罐中Kringle 5蛋白的最终表达量为0.59g/L.     

食用色素红曲红的急性和亚慢性经口毒性研究

目的 通过开展食用色素红曲红的急性和亚慢性经口毒性试验,探讨其食用安全性,为红曲红的风险评估和标准修订提供依据。方法 急性经口毒性试验采用限量法。KM种小鼠24 h内一次经口灌胃给予10 g/kg·bw红曲红,观察动物的中毒体征和死亡情况;亚慢性经口毒性试验采用148只SPF级SD种大鼠(雌雄各半),主试验组大鼠经口灌胃给予0.50、1.08、2.32和5.00 g/kg·bw的红曲红90 d,中期和恢复期卫星组大鼠分别经口灌胃给予5.00 g/kg·bw的红曲红45 d, 90 d后继续观察28 d。结果 急性经口毒性试验期间未出现动物中毒反应和死亡,获得雌性和雄性小鼠经口LD50>10 g/kg·bw;亚慢性经口毒性试验期间无动物死亡,红曲红对大鼠一般临床观察、体重和摄食、血液学、血清生化、尿常规、尿生化及病理学指标均无明显影响。结论 红曲红急性经口毒性分级为实际无毒;亚慢性经口毒性雌性和雄性大鼠未观察到有害作用剂量为5.00 g/kg·bw(本试验最高剂量)。     

CLP配方的急性毒性及其对小鼠急性酒精性肝损伤的保护作用

目的研究CLP配方的急性经口毒性及对急性酒精性肝损伤的辅助保护作用。方法采用Wistar大鼠进行急性经口毒性试验研究;急性酒精性肝损伤的辅助保护作用试验采用50%酒精诱导昆明小鼠建立肝损伤模型,随机分为阴性对照组、模型对照组及低、中、高3个剂量组,连续给样30 d,末次给样后,各剂量组及模型对照组给予酒精造模。禁食16 h后,取样,对小鼠试验前后体重与肝脏重量相关指标进行分析,测定小鼠肝组织中还原型谷胱甘肽(GSH)、甘油三酯(TG)、丙二醛(MDA)的含量,同时对小鼠肝脏进行组织病理学检查。结果各剂量组试验前后体重与模型对照组比较差异无显著性;CLP配方能升高还原型GSH含量,降低TG含量;同时能减轻肝损伤和肝细胞脂肪变性。结论 CLP配方对Wistar大鼠急性经口毒性LD50大于20000 mg/kg·BW,属无毒级;CLP配方对小鼠急性酒精性肝损伤有辅助保护作用。     

两种动物品系与方法比较硝酸镧的急性经口毒性

目的 以近交系C57小鼠与远交系KM小鼠为研究对象,应用霍恩氏(Horn)法与上-下法(up-down procedure,UDP)的改良方案,比较硝酸镧(Ⅲ)六水合物的急性经口毒性.方法 参照国内外急性经口毒性试验标准或指南中Horn法与UDP法的设计与计算原理,分别选用90只C57小鼠与KM小鼠(雌雄各半)进行随机...     

抗菌肽Cecropin B-肿瘤血管生长抑制因子 Kringle5融合蛋白表达载体的构建及其表达

通过重叠区扩增法人工合成抗菌肽Cecropin B(CB)基因,从ThioA/L15-7上卸下肿瘤血管生长抑制因子Kringle5(K5)基因,将两者按正确的阅读框架融合并定向克隆至大肠杆菌高效表达载体pET32a上,构建成pET32a/CB-K5重组载体。重组载体转化BL21(DE3)细胞后,提取质粒进行PCR鉴定和序列分析,证明重组质粒pET32a/CB-K5阅读框架和融合基因序列均与预期相符。实验结果显示,成功地构建了抗菌肽CecropinB(CB)和肿瘤血管生长抑制因子Kringle5(K5)融合蛋白的表达载体pET32a/CB-K5。转化E.coliBL21(DE3),SDS-PAGE分析显示,在IPTG诱导下,融合蛋白以包涵体的形式在重组转化菌株中获得高效表达,表达量达到菌体总蛋白的50%。     

The international validation of a fixed-dose procedure as an alternative to the classical LD50 test

In an international study involving 33 laboratories in 11 countries, the acute oral toxicity to the rat of 20 substances and preparations was evaluated using a fixed-dose procedure and the results compared with those obtained for the test materials using the classical LD50 test. The study has shown that the fixed-dose approach to acute oral toxicity testing: (1) produces consistent results that are not substantially affected by inter-laboratory variations; (2) provides adequate information for risk assessment purposes on signs of toxicity, including their nature, time to onset, duration and outcome; (3) uses fewer animals than the current internationally agreed OECD procedure (Guideline 401-revised); (4) subjects animals to less pain and distress than the classical LD50 test and causes less compound-related mortality; and (5) enables substances and preparations to be ranked according to the EEC classification system on the basis of their acute oral toxicity, such ranking being compatible with that allocated by the results of classical LD50 studies.     

拟原白头翁素A毒性试验研究

目的为了对拟原白头翁素Ａ进行初步毒理学安全性评价。方法采用大鼠急性经口和经皮毒性试验，大鼠皮肤和大白兔眼粘膜刺激试验，Ａｍｅｓ试验，小鼠骨髓微核试验，小鼠睾丸初级精母细胞染色体畸变试验。结果大鼠急性经口ＬＤ５０４．６７３９（ＩＣ４．０６１４～５．３７８７）ｇ·ｋｇ－１，经皮ＬＤ５０＞２．０ｇ·ｋｇ－１，根据化合物毒性分级为低毒；对大鼠皮肤无刺激作用，对大白兔眼粘膜有中等刺激；Ａｍｅｓ试验、小鼠骨髓微核试验和小鼠睾丸初级精母细胞染色体畸变试验阴性。结论拟原白头翁素Ａ为低毒、致突变试验阴性的化合物。     

Oral acute toxic class method: a successful alternative to the oral LD50 test

The oral acute toxic class method (ATC method) was developed as an alternative to replace the oral LD50 test. The ATC method is a sequential testing procedure using only three animals of one sex per step at any of the defined dose levels. Depending on the mortality rate three but never more than six animals are used per dose level. This approach results in the reduction of numbers of animals used in comparison to the LD50 test by 40-70%. The principle of the oral ATC method is based on the Probit model and it was first evaluated on a biometric basis before a national and subsequently an international ring study were conducted. The results demonstrated an excellent agreement between the toxicity and the animal numbers predicted biometrically and observed in the validation studies. The oral ATC method was adopted as an official test guideline by OECD in 1996 and was slightly amended in 2001. The ATC method has been successfully used in Germany and in 2003 >85% of all tests on acute oral toxicity testing was conducted as oral ATC tests. In member states of the European Union the ATC method is used in the range of 50% of all tests conducted. Meanwhile the oral LD50 test has been deleted by OECD, by the European Union and by the USA, making the use of alternatives to the oral LD50 test mandatory.     

鼻渊灵冲剂的毒性实验研究

鼻渊灵口服（ｉｇ）测不出ＬＤ_(５０)，小鼠ｉｇ最大耐受量为１１２．５ｇ／ｋｇ；小鼠腹腔注射，ＬＤ-(５０)为４２．７ｇ／ｋｇ。长期毒性试验表明，大鼠分别ｉｇ鼻渊灵１５、３０、４５ｇ／ｋｇ，连续用药１２周，血液常规、生化指标及脏器系数均在正常范围内。结果提示，鼻渊灵ｉｇ毒性低，长期应用安全可靠。     

Comparison of the up-and-down method and the fixed-dose procedure for acute oral toxicity testing

The acute oral toxicity data for 10 compounds, generated by using two alternative methods in rats, the up-and-down method and the fixed-dose procedure, were compared with those obtained from the classical LD50 test. In this evaluation, both alternative methods offered a reduction in animal use, while providing adequate information to rank the compounds according to the EEC classification for acute oral toxicity. In addition to the ranking, these alternative methods also provided useful information on signs of toxicity and gross autopsy findings, although the results varied depending on the method used. Of the three methods, the up-and-down method required the fewest animals. Although the up-and-down method used only females, the LD50 values obtained were in good agreement with those obtained by the classical method, which used both sexes. It is concluded that the up-and-down method and the fixed-dose procedure are acceptable alternative methods to the classical LD50 test, and the choice of method depends on the type of toxicity information required.     

Toxicity of binary chemical munition destruction products: methylphosphonic acid, methylphosphinic acid, 2-diisopropylaminoethanol, DF neutralent, and QL neutralent

This paper reports the toxicity and environmental impact of neutralents produced from the hydrolysis of binary chemical agent precursor chemicals DF (methylphosphonic difluoride) and QL (2-[bis(1-methylethyl)amino]ethyl ethyl methylphosphonite). Following a literature review of the neutralent mixtures and constituents, basic toxicity tests were conducted to fill data gaps, including acute oral and dermal median lethal dose assays, the Ames mutagenicity test, and ecotoxicity tests. For methylphosphonic acid (MPA), a major constituent of DF neutralent, the acute oral LD(50) in the Sprague-Dawley rat was measured at 1888 mg/kg, and the Ames test using typical tester strains of Salmonella typhimurium and Escherichia coli was negative. The 48-h LC(50) values for pH-adjusted DF neutralent with Daphnia magna and Cyprinodon variegatus were > 2500 mg/L and 1593 mg/L, respectively. The acute oral LD(50) values in the rat for QL neutralent constituents methylphosphinic acid (MP) and 2-diisopropylaminoethanol (KB) were both determined to be 940 mg/kg, and the Ames test was negative for both. Good Laboratory Practice (GLP)-compliant ecotoxicity tests for MP and KB gave 48-h D. magna EC(50) values of 6.8 mg/L and 83 mg/L, respectively. GLP-compliant 96-h C. variegatus assays on MP and KB gave LC(50) values of 73 and 252 mg/L, respectively, and NOEC values of 22 and 108 mg/L. QL neutralent LD(50) values for acute oral and dermal toxicity tests were both > 5000 mg/kg, and the 48-h LD(50) values for D. magna and C. variegatus were 249 and 2500 mg/L, respectively. Using these data, the overall toxicity of the neutralents was assessed.     

羧甲基壳多糖毒理学研究

羧甲基壳多糖的毒理学研究，包括急性皮肤毒性，急性经口毒性，长期经口毒性。研究结果表明：羧甲基壳多糖属安全无毒性质。对小白鼠急性皮肤毒性ＬＤ５０〉２２００ｍｇ／ｋｇ，急性经口毒性，ＬＤ５０〉５００１ｍｇ／ｋｇ。长期经口毒性实验，采用高，中，低３个剂量组，对大白鼠体重增长百发率，血象，内脏外观，肝肾功能及肝，肾组织学检查，均未发现异常。     

Statistical evaluation of the revised fixed-dose procedure

The fixed-dose procedure (FDP) was introduced as OECD Test Guideline 420 in 1992, as an alternative to the conventional median lethal dose (LD50) test for the assessment of acute oral toxicity (OECD Test Guideline 401). The FDP uses fewer animals and causes less suffering than the conventional test, while providing information on the acute toxicity to allow substances to be ranked according to the EU hazard classification system. Recently the FDP has been revised, with the aim of providing further reductions and refinements, and classification according to the criteria of the Globally Harmonized Hazard Classification and Labelling scheme (GHS). This paper describes the revised FDP and analyses its properties, as determined by a statistical modelling approach. The analysis shows that the revised FDP classifies substances for acute oral toxicity generally in the same, or a more stringent, hazard class as that based on the LD50 value, according to either the GHS or the EU classification scheme. The likelihood of achieving the same classification is greatest for substances with a steep dose-response curve and median toxic dose (TD50) close to the LD50. The revised FDP usually requires five or six animals with two or fewer dying as a result of treatment in most cases.     

Acute toxicity of aversectin C : various routes of administration and dosage forms

The acute oral, cutaneous, and inhalation toxicity of aversectin C was studied on white unbred rats and mice. The compound was less toxic for rats than for mice, the LD50 for oral administration being 90 and 33 mg/kg, respectively. Aversectin C exhibited a maximum acute toxicity upon the inhalation in rats (LD50 = 40 mg/kg), while a minimum toxicity level was observed for the cutaneous application in rats (1700 mg/kg).     

Structure-toxicity relationship of monoketones

The structure-toxicity relationship of monoketones was examined in mice. In all test compounds, the acute oral toxicity (LD50) was determined under two conditions; control LD50 (LD50-cont.) and carbon tetrachloride (CCl4)-pretreated LD50 (LD50CCl4). LD50-cont. was larger than LD50-CCl4 in all test compounds, the difference being greater in compounds with longer carbon chains than in those with shorter chains. Both LD50-cont. and LD50-CCl4 were parabolic functions of the partition coefficient, log P, i.e., log (1/LD50-cont.) = -0.120(log P)2 + 0.417log P - 1.734 and log(1/LD50-CCl4) = -0.103(log P)2 + 0.462log P - 1.660, and both equations were statistically significant (P less than 0.01). The log Po values were 1.74 for the former and 2.24 for the latter.