霞浦某市场虾苗酱的致突变性试验研究

目的 通过细菌回复突变试验和体外哺乳类细胞HGPRT基因突变试验探索虾苗酱是否存在致突变性。方法 细菌回复突变试验选用经鉴定符合要求的鼠伤寒沙门氏菌TA97a、TA98、TA100、TA1535及大肠杆菌,试验设置5 000、1 580、500、158、50μg/皿剂量组进行试验。体外哺乳类细胞HGPRT基因突变试验采用MTT法检测虾苗酱对V79细胞的毒性,确定以5 000μg/mL为最高剂量进行V79细胞HGPRT基因突变试验。结果 各剂量组虾苗酱对测试菌株诱发产生的回变菌落数均未超过溶剂对照组回变菌落数的2倍;各剂量组虾苗酱的V79细胞集落突变率与溶剂对照组差异无统计学意义。结论 在本试验条件下,霞浦虾苗酱无细菌回复突变性、无基因突变作用。     

4种植物型染发剂的遗传毒性研究

目的了解4种植物型染发剂的遗传毒性作用,掌握其潜在的健康危害。方法通过鼠伤寒沙门氏菌回复突变试验(Ames试验)、CHL细胞染色体畸变试验和V79细胞HGPRT基因突变试验,检测4种染发剂对原核细胞和真核细胞在基因水平和染色体水平的诱变性。结果 Ames试验结果显示,1号和2号染发剂在高剂量5000μg/皿时,在添加和未添加代谢活化系统条件下,TA97、TA98和TA100的菌落回变数异常增加,结果为阳性;3号和4号染发剂在添加和未添加代谢活化系统条件下,各剂量组菌落回变数均在正常范围内,结果为阴性。与阴性对照组比较,4种植物型染发剂各剂量组均未引起CHL细胞染色体畸变率和V79细胞HGPRT基因突变频率明显增加(P>0.05),结果均为阴性。结论在本实验条件下,3号和4号染发剂未显示遗传毒性。1号和2号染发剂尽管在真核细胞水平的试验结果为阴性,但仍需十分谨慎评价其潜在的健康危害。     

细菌回复突变试验在聚乙烯醇栓塞微球材料评价中的应用

目的：通过细菌回复突变试验（Ames试验）评价聚乙烯醇栓塞微球潜在的遗传毒性，从而来评价该材料的潜在致突变性。方法：选择生理盐水（0.9%氯化钠水溶液）、二甲基亚砜（DMSO）和含10%血清培养基（R10）三种介质对试验样品进行浸提，采用平板掺入法计数鼠伤寒沙门氏菌组氨酸营养缺陷型菌株TA97a、TA98、TA100及TA102在活化和非活化条件下的细菌回复突变菌落数，以检测其致突变性。结果：阴性对照组和阳性对照组的试验结果成立。聚乙烯醇栓塞微球采用生理盐水、DMSO和R10浸提后，浸提液对试验所用菌株回复突变数均未超过阴性对照的2倍。结论：在本次试验条件下，栓塞微球浸提液均对鼠伤寒沙门氏菌营养缺陷型菌株TA97a、TA98、TA100、TA102无诱变性。     

1，1-二氨基2，2-二硝基乙烯对哺乳动物细胞致突变性研究

在非代谢活化（-S9）和代谢活化（+S9）条件下，以1，1-二氨基-2，2-二硝基乙烯（FOX-7）处理小鼠淋巴瘤L5178Y细胞和中国仓鼠肺CHL细胞，进行体外哺乳动物细胞TK基因突变试验和染色体畸变试验。结果显示，分别以500、250、125 μg/ml浓度的FOX-7处理L5178Y细胞，各剂量组突变频率与溶剂对照组相比，差异有统计学意义（P<0.05），且具有剂量相关性；分别以300、200、100 μg/ml浓度FOX-7处理CHL细胞，各剂量组染色体畸变细胞率与溶剂对照组比较，差异无统计学意义（P>0.05）。提示FOX-7可能对小鼠淋巴瘤L5178Y细胞TK基因具有致突变性，对中国仓鼠肺CHL细胞染色体无致突变性。     

脱细胞异体真皮基质的遗传毒性研究

目的:检测医疗器械注册产品脱细胞异体真皮基质的遗传毒性.材料与方法:以产品浸提液为供试液,采用细菌回复突变试验(Ames 试验) 和小鼠淋巴瘤细胞试验(MLA),在非代谢活化(-S9) 和代谢活化(+S9) 条件下检测.Ames 试验采用平板掺入法,检测了50 、100 、200 μl/ 皿3个剂量组诱发鼠伤寒沙门氏菌TA98 、TA100 、TA1535 、TA1537 及大肠杆菌WP2 uvrA 的回变菌落数;MLA 检测了终浓度2.5% 、5% 、10% (V/V) 3 个浓度组,在+S9/3 h、-S9/3 h 、-S9/24 h 三种处理条件下诱发的L5178Y 细胞TK 基因突变频率(MF) 和小集落突变百分率(SC%).结果:未经复水处理所得样品浸提液诱发五株菌的回变菌落数,对比氯化钠注射液溶媒(阴性)对照组均无明显增加;-S9/3 h 处理条件下诱发的MF 明显增加(P<0.01),且具有浓度相关性;复水处理后所得样品浸提液,在-S9/3 h 、24 h 条件下诱发的MF 均无明显增加.结论:脱细胞异体真皮基质对测试菌株his-/trp-无明显诱变作用,未经复水处理时在-S9 条件下对测试细胞tk-+/-及染色体具明显损伤作用,经复水处理后无明显损伤作用.产品在非代谢活化条件下表现出的阳性结果可能与其附加的冷冻保护剂有关.     

生川乌提取物遗传毒性研究

[目的]选用目前新药遗传毒性评价中推荐使用的3种试验方法(小鼠骨髓微核试验、鼠伤寒沙门氏菌回复突变试验、体外CHL细胞染色体畸变试验)研究生川乌的遗传毒性.[方法]小鼠骨髓微核试验设3个生川鸟提取物给药剂量组(26.0、13.0、6.5 g生药/kg)、阴性对照组和阳性对照组(环磷酰胺40 mg/kg).Ames试验选用两种方法:直接法和代谢活化法.试验菌株为鼠伤寒沙门氏组氨酸缺陷型菌株TA97、TA98、TA100、TA102、TA1535,生川乌提取物在试验中设6个剂量组(5.000、2.500、11250、0.625、0.313、0.156 mg生药/皿),同时设自发回复突变对照组、阳性对照组.体外CHL细胞染色体畸变试验也选用两种方法:直接法和代谢活化法.试验设5个生川乌提取物浓度组(5.000、2.500、1.250、0.625、0.313 mg生药/ml),阴性对照及阳性对照组.[结果]小鼠骨髓微核试验生川乌提取物各剂量组小鼠骨髓多染红细胞微核率与阴性对照组相比,差异无统计学意义(P>0.05).Ames试验生川乌提取物在加或不加肝微粒体酶(S9)时均未见引起TA97、TA98、TA100、TA102、TA1535试验菌株基因突变,即Ames试验阴性,试验重复一次,所得结论相同.体外CHL细胞染色体畸变试验生川乌提取物在加或不加S9时均未引起CHL细胞的染色体畸变,试验结果为阴性,试验重复1次,所得结论相同.[结论]在本实验室条件下,生川乌提取物在小鼠骨髓微核试验、Ames试验及体外CHL细胞染色体畸变试验中均为阴性结果,故认为在本实验条件下,生川乌提取物无遗传毒性.     

1,2,4-三唑-5-酮的遗传毒性研究

为探讨1,2,4-三唑-5-酮(TO)的遗传毒性,采用鼠伤寒沙门菌回复突变试验(Ames试验),计算回复突变菌落数;小鼠骨髓嗜多染红细胞微核试验,计算小鼠股骨骨髓细胞微核率;中国仓鼠肺细胞(CHL)体外染色体畸变试验,观察染色体结构畸变类型并计算畸变率。结果显示,TO对TA97a、TA98、TA100、TA102和TA1535 5种鼠伤寒沙门菌无致突变性,对哺乳动物培养细胞染色体无致畸作用,也无诱发骨髓嗜多染红细胞微核形成的作用,TO不具有潜在的遗传毒性。     

盐附子遗传毒性研究

[目的]选用目前新药遗传毒性评价中推荐使用的3种试验方法(小鼠骨髓微核试验、鼠伤寒沙门氏菌回复突变试验、体外CHL细胞染色体畸变试验)研究盐附子的遗传毒性. [方法]小鼠骨髓微核试验设3个盐附子给药剂量组(15.45、7.73、3.86 g生药/kg)、阴性对照组和阳性对照组(环磷酰胺40 mg/kg).Ames试验选用两种方法:直接法和代谢活化法.试验菌株为鼠伤寒沙门氏组氨酸缺陷型菌株TA97、TA98、TA100、TA102、TA1535,盐附子在试验中设6个剂量组(5.000、2.500、1.250、0.625、0313、0.156 mg生药/皿),同时设自发回复突变对照组、阳性对照组.体外CHL细胞染色体畸变试验也选用两种方法:直接法和代谢活化法.盐附子不加S9时处理终浓度为5、2.5、1.25、0.625、0.313 mg生药/ml,加S9时处理终浓度为2.5、1.25、0.625、0.313、0.156 mg生药/ml. [结果]小鼠骨髓微核试验盐附子各剂量组小鼠骨髓多染红细胞微核率与阴性对照组相比,差异无统计学意义(P>0.05).Ames试验盐附子在加或不加肝微粒体酶(S9)时均未见引起TA97、TA98、TA100、TA102、TA1535试验菌株基因突变,即Ames试验阴性,试验重复一次,所得结论相同.体外CHL细胞染色体畸变试验盐附子在加或不加S9时均未引起CHL细胞的染色体畸变,试验结果为阴性,试验重复一次,所得结果相同.[结论]在实验室条件下,盐附子在小鼠骨髓微核试验、Ames试验及体外CHL细胞染色体畸变试验中均为阴性结果,故认为盐附子无遗传毒性.     

抗菌脂肽对体外哺乳动物细胞遗传毒性的研究

目的研究抗菌脂肽对体外哺乳动物细胞的遗传毒性。方法根据细胞毒性试验结果,在存在/不存在代谢活化系统条件下:分别设86、43、22mg/L和36、18、9mg/L 3个剂量组,以抗菌脂肽作用中国地鼠肺成纤维细胞(CHL),分析CHL染色体致畸变率;分别设7.5、15、30、60、120mg/L和2.5、5、10、20、40mg/L 5个剂量组,以抗菌脂肽作用中国仓鼠肺成纤维细胞(V79),通过HGPRT基因突变试验,计算相对集落形成率,评估抗菌脂肽对体外哺乳动物细胞的致突变性。结果在存在/不存在代谢活化系统条件下,抗菌脂肽对CHL细胞染色体畸变率为2.0%~4.0%;作用V79细胞后,MF均11.0×10-6;均低于阳性对照,差异均有统计学意义(P值均0.05)。以上指标与阴性对照组差异均无统计学意义(P值均0.05)。结论在体外试验条件下,抗菌脂肽对哺乳动物细胞无致突变性或潜在的遗传毒性。     

克菌丹Ames试验的研究

目的研究克菌丹对Ames试验菌株的致突变性。方法取农药克菌丹,按照GB15670—1995《农药登记毒理学试验方法》中的平板掺入法进行试验和结果评价。结果在不加S-9代谢活化系统条件下,TA97、TA100试验菌株各剂量组(10～50μg/皿)回复突变菌落数随剂量增高而增多,均大于自发回复突变菌落数的2倍,存在剂量-反应关系,且有重复性,为阳性反应;在加S-9代谢活化系统条件下,TA97、TA98、TA100试验菌株中、高剂量组(25～50μg/皿)回复突变菌落数随剂量增高而增多,均大于自发回复突变菌落数的2倍,存在剂量-反应关系,且有重复性,为阳性反应。结论在本试验条件下,克菌丹对鼠伤寒沙门氏菌回复突变试验(Ames试验)为阳性。     

贵州铁皮石斛细菌回复突变试验研究

目的对贵州铁皮石斛进行细菌回复突变试验,评价其是否具有潜在的致突变性。方法采用生物学性状经鉴定遗传学特征符合要求的鼠伤寒沙门菌突变型TA97a、TA98、TA100、TA102、TA1535进行试验,用肝匀浆作为体外代谢活化系统(+/-s9),同时设自发回变、溶剂对照组和阳性对照组。试验分两次进行,每个剂量组及对照组均做3个平行样,在37t培养48 h以上,计数每皿回变菌落数。结果两次试验结果显示,无论在+S9和-S9的试验条件下,空白对照组、溶剂对照组的自发回变菌落数均在标准范围内,各剂量组的TA97a、TA98、TA100、TA102、TA1535回变菌落平均数均未超过阴性对照组回变菌落平均数的2倍,亦无剂量相关性,细菌回复突变试验结果为阴性。结论在本试验剂量范围内,铁皮石斛细菌回复突变试验结果为阴性,提示贵州铁皮石斛无致突变性。     

对二氯苯致突变性的实验研究

目的　研究对二氯苯的致突变性。方法　Ames试验用TA97、TA98、TA10 0和TA10 2菌株 ,加与不加S 9试验 (剂量设为 0 2 5、0 5、1 0、2 0、5 0mg/皿 ) ;小鼠骨髓多染红细胞微核试验 (剂量设为 10 0、2 0 0、40 0、80 0mg/kg)和小鼠睾丸细胞染色体畸变试验 (剂量设为 2 5 0、5 0 0、10 0 0mg/kg)。结果　Ames试验中各测试浓度的回变菌落数均未超过自然回变菌落数 2倍 ;小鼠骨髓多染红细胞微核试验和小鼠睾丸细胞染色体畸变试验 ,各剂量组和阴性对照组差异无显著性 (P >0 0 5 )。结论　该试验范围内 ,未见对二氯苯有致突变性。     

90%单甲脒盐酸盐原药的TK基因突变试验

目的研究90%单甲脒盐酸盐原药的致突变性。方法在活化和非活化条件下,采用小鼠淋巴瘤细胞株(L5178Y-3.7.2C)进行TK基因突变试验,分析不同剂量90%单甲脒盐酸盐原药(0.078～5.0 mg/ml)的致突变性。结果在有和无体外代谢活化系统S9时,1.25、0.625、0.313、0.156 mg/ml剂量组的90%单甲脒盐酸盐原药TK位点突变频率稍有增加,但与溶剂对照组比较,均无统计学意义(P>0.05)。结论在本体外哺乳动物细胞基因突变试验剂量范围内,未见90%单甲脒盐酸盐原药具有致突变性。     

Mutagenicity studies in a tyre plant: in vitro activity of workers' urinary concentrates and raw materials

The possible contribution to urinary mutagenicity of occupational exposures in the rubber industry was studied by assaying the urine concentrates of 72 workmen (44 smokers) employed in a tyre plant. Twenty three clerks (16 smokers) engaged in the administrative department of the same factory served as presumptive unexposed controls. XAD-2 resin concentrates of urine samples were assayed in the plate incorporation test and in the microtitre fluctuation assay with Salmonella typhimurium strains TA1535, TA98, and TA100. Furthermore, the in vitro mutagenicity of the major raw materials in use at the plant was determined in the plate incorporation assay with S typhimurium strains TA1535, TA1537, TA98, and TA100. The results obtained from the urinary mutagenicity study show that smoking habits, but not occupation, were statistically significantly related to the appearance of a urinary mutagenicity that was detectable with strain TA98. A possible synergistic effect of occupation with smoking was observed among tyre builders who were also smokers. The study of the raw materials showed that three technical grade materials were weakly active as mutagens in strain TA98 in the absence (poly-p-dinitrosobenzene) or in the presence of metabolic activation (mixed diaryl-p-phenylendiamines and tetramethyltiuram disulphide). The latter chemical was also weakly active in strain TA100.     

Mutagenicity studies in a tyre plant: in vitro activity of workers'' urinary concentrates and raw materials.

The possible contribution to urinary mutagenicity of occupational exposures in the rubber industry was studied by assaying the urine concentrates of 72 workmen (44 smokers) employed in a tyre plant. Twenty three clerks (16 smokers) engaged in the administrative department of the same factory served as presumptive unexposed controls. XAD-2 resin concentrates of urine samples were assayed in the plate incorporation test and in the microtitre fluctuation assay with Salmonella typhimurium strains TA1535, TA98, and TA100. Furthermore, the in vitro mutagenicity of the major raw materials in use at the plant was determined in the plate incorporation assay with S typhimurium strains TA1535, TA1537, TA98, and TA100. The results obtained from the urinary mutagenicity study show that smoking habits, but not occupation, were statistically significantly related to the appearance of a urinary mutagenicity that was detectable with strain TA98. A possible synergistic effect of occupation with smoking was observed among tyre builders who were also smokers. The study of the raw materials showed that three technical grade materials were weakly active as mutagens in strain TA98 in the absence (poly-p-dinitrosobenzene) or in the presence of metabolic activation (mixed diaryl-p-phenylendiamines and tetramethyltiuram disulphide). The latter chemical was also weakly active in strain TA100.     

Assessment of the mutagenic potency of sewage sludges contaminated with polycyclic aromatic hydrocarbons by an Ames sludges for fluctuation assay

The mutagenicity of crude extracts and subfractions of two samples of a reference sewage sludge material and two sewage sludges from two wastewater treatment plants (WWTP), one urban and the other one urban mixed with industrial, was assessed using an Ames fluctuation assay based on 384-well microtiter plates with liquid cultures. Crude extracts of sludges were obtained by ultrasonic extraction with dichloromethane/methanol, and further column fractionation yielded two fractions, one of which containing mutagenic polycyclic aromatic hydrocarbons (PAHs). Quantitative analysis performed by gas chromatography-mass spectrometry gave sum concentrations of the 16 PAHs listed as priority pollutants by the U.S. Environmental Protection Agency at levels between 1,305 and 2,442 microg/kg. Subjecting crude extracts and column fractions to the mutagenicity assay with Salmonella strains TA98 and TA 100 provided good qualitative correlation between the presence of mutagenic PAH and the induction of gene mutations. In general, the crude extracts and the PAH-fractions induced positive responses in the assay with both bacterial strains on metabolic activation by S9 rat-liver homogenate, whereas direct-acting mutagens were not detectable. In the assay with the real sludge samples of two different WWTPs, TA98 proved to be more sensitive than TA100; however, similar sensitivities of the tester strains were observed for two reference sewage sludge materials of the same origin. The outcomes of the Ames fluctuation assay demonstrated its performance as a cost-effective and relatively rapid screening tool to assess the genotoxic potential of complex environmental samples.     

碳纳米管的Ames试验研究

目的应用Ames试验系统观察多壁碳纳米管的诱变活性。方法选用TA97、TA98、TA100、TA102标准测试菌株,采用标准平板掺入法检测多壁碳纳米管的诱变活性。加S9混合液作为体外代谢活化系统。结果多壁碳纳米管在S9活化和非活化两种测试条件下,1～500μg/皿浓度范围,诱发TA98、TA100两种测试菌株回变菌落数与阴性对照组相比无明显增加,在1～10μg/皿浓度范围,诱发TA97和TA102产生的回变菌落数与阴性对照相比无明显增加,在10～500μg/皿测试浓度范围,对TA97和TA102产生的回变菌落数与阴性对照相比均达到阴性对照两倍以上。结论多壁碳纳米管具有碱基置换及移码型的直接诱变特性和间接诱变特性。     

Carcinogenicity of extract of airborne particles using newborn mice and comparative study of carcinogenic and mutagenic effect of the extract

An organic extract of airborne particles collected in Tokyo and its fractions (neutral, acidic, and basic) were investigated in Ames Salmonella assays for mutagenicity and in newborn mice for carcinogenicity. Mutagenicity to TA100 and TA98 strains was detected in the whole extract, the neutral, the acidic and the basic fractions with and without metabolic activation. In the carcinogenicity test, the incidence of lung tumor was as follows: whole extract, 4/25; neutral fraction, 7/25; acidic fraction, 0/20; basic fraction, 1/11; vehicle, 2/39; and uninjected, 3/47. The neutral fraction of the extract of the airborne particles showed highly potent mutagenicity and a high incidence of lung tumors in mice.     

3,4-双（4＇-氨基呋咱基-3′）氧化呋咱的Ames试验研究

目的应用Ames试验评价3,4-双（4′-氨基呋咱基-3′）氧化呋咱（DATF）的致突变作用。方法选择组氨酸营养缺陷型鼠伤寒沙门杆菌TA97a、TA98、TAl00和TAl02标准测试菌株,DATF设2 500、1 000、500、200、100μg/皿共5个浓度的剂量组,同时设溶剂对照和阳性对照,在有和无S9代谢活化剂下采用平板掺入法检测DATF的诱变活性。结果 DATF在S9活化和非活化两种测试条件下,诱发TA98、TA100、TA102 3种菌株回变菌落数与阴性对照组相比无明显增加;在500和1 000μg/皿2个剂量下诱发TA97a产生的回变菌落数与阴性对照相比均达到阴性对照2倍以上,并且存在剂量-效应关系。结论 DATF对鼠伤寒沙门杆菌TA97a具有一定的诱变特性。     

The results of microbial mutation test for forty-three industrial chemicals

The mutagenicity of 43 industrial chemicals in Salmonella typhimurium (TA98, TA100, TA1535, TA1537, and TA1538) and Escherichia coli (WP2uvrA) was examined. The output of these chemicals in Japan is more than a million kilograms per year. The mutation test was carried out under the condition of absence and presence of rat microsomal activation. Two chemicals, hexamethylenetetramine and 4,4'-methylenediphenyldiisocyanate, showed mutagenic activity in S. typhimurium TA98 and TA100 by metabolic activation. Hexamethylenetetramine also showed mutagenic activity in TA98 without microsomal activation. No mutagenic activity was observed in the 41 chemicals including 4 volatile and gaseous compounds.