盐酸川丁特罗遗传毒性研究

目的：研究盐酸川丁特罗（SPFF）的遗传毒性。方法：采用Ames试验、哺乳动物培养细胞染色体畸变试验、小鼠微核试验，考察SPFF对鼠伤寒沙门菌和细胞的遗传毒性。结果：经Ames试验、CHL细胞染色体畸变试验、小鼠微核试验，SPFF的结果均呈阴性。结论：SPFF无遗传毒性。     

益母草碱对胚胎-胎仔发育毒性和遗传毒性的评价

目的 检测益母草碱的发育毒性和遗传毒性。方法 在SD孕鼠妊娠第6~15天经口灌胃给予500、1 000和2 000 mg/kg体重的益母草碱,同时设溶媒对照组,经口灌胃0.5% CMC-Na溶液。妊娠第20天剖杀孕鼠,分析其生殖毒性。分别采用反映基因突变的鼠伤寒沙门菌回复突变试验（Ames试验）、反映染色体畸变的细胞染色体畸变试验（体外培养CHO）和ICR小鼠骨髓微核试验（体内）检测益母草碱的遗传毒性。结果 在500、1 000和2 000 mg/kg剂量益母草碱的作用下孕鼠的增重与对照组相比,差异均无统计学意义;各受试剂量组孕鼠各项指标与对照组相比,差异均无统计学意义;各剂量组胎鼠各类指标与溶媒对照组相比,无明显差异。Ames试验结果提示：在0.5、5、50、500、5 000 μg/皿受试剂量下,在有或无代谢活化S9系统时,与溶媒对照组相比,受试物对组氨酸缺陷型鼠伤寒沙门菌（TA97、TA98、TA100、TA102及TA1535）所诱发的回复突变菌落数均相近。染色体畸变试验结果显示：250、500和1 000 μg/ml 3个剂量的受试物,对有或无代谢活化系统S9培养的CHO细胞的染色体畸变率无明显影响。微核试验显示100、500和2 000 mg/kg各个剂量组对ICR小鼠的微核诱发率与溶媒对照组比较均无显著差异（P>0.05）。结论 益母草碱在500、1 000和2 000 mg/kg剂量下未观察到明显的母体毒性、胚胎毒性、胎儿毒性和致畸作用。益母草碱对鼠伤寒沙门菌无致突变性,对哺乳动物培养细胞的染色体无致畸变作用,对ICR小鼠无诱发骨髓嗜多染红细胞微核的效应。上述结果表明在本试验条件下,益母草碱无发育和遗传毒性。     

富马酸泰诺福韦双特戊酯的遗传毒性研究

目的 检测富马酸泰诺福韦双特戊酯(TDF)的遗传毒性,为临床用药提供理论依据.方法 应用鼠伤寒沙门细菌回复突变试验(Ames试验)、体外培养中国仓鼠肺成纤维细胞(CHL)细胞染色体畸变试验和小鼠骨髓微核试验检测该药物的遗传毒性.结果 该药物对鼠伤寒沙门菌无致突变性,对体外培养CHL细胞染色体无致畸变作用,对昆明小鼠无诱发骨髓嗜多染红细胞微核的效应,三个试验结果均呈阴性.结论 TDF不具有遗传毒性.     

聚乙二醇修饰降纤酶的遗传毒性评价

目的检测聚乙二醇修饰降纤酶的遗传毒性。方法应用鼠伤寒沙门菌回复突变试验(Ames试验)、体外培养CHO细胞染色体畸变试验和小鼠骨髓微核试验检测聚乙二醇修饰降纤酶的遗传毒性。结果 Ames试验结果显示每平皿100、20、4、0.8、0.16 U各个剂量组,在加或不加S9代谢活化系统时对组氨酸缺陷型鼠伤寒沙门菌TA97、TA98、TA100、TA102及TA1535所诱发的回复突变菌落数均与溶剂对照的突变菌落数相近。体外培养CHO细胞染色体畸变试验结果显示2.5、5.0和10.0 U.mL-1各个剂量组在加S9代谢活化系统于24 h和不加S9代谢活化系统于24 h、48 h培养的CHO细胞染色体畸变率与溶剂对照组比较均无显著差异(P>0.05)。小鼠骨髓微核试验显示425、850、1700 U.kg-1各个剂量组对ICR小鼠的微核诱发率与溶剂对照组比较均无显著差异(P>0.05)。结论聚乙二醇修饰降纤酶对鼠伤寒沙门菌无致突变性,对哺乳动物培养细胞的染色体无致畸变作用,对ICR小鼠无诱发骨髓嗜多染红细胞微核的效应。表明聚乙二醇修饰降纤酶在本实验条件下无遗传毒性。     

中药浮海石致突变的实验研究

目的研究中药浮海石致突变性。方法本试验采用鼠伤寒沙门菌回复突变试验(Ames试验),哺乳动物培养细胞(CHL)染色体畸变试验和小鼠骨髓细胞微核试验观察了浮海石的致突变作用。结果浮海石生理盐水浸提液0.5~5000μg/皿剂量下Ames试验结果阴性;250~1000μg/mL剂量下对CHL细胞无损伤作用,10~40g/kg剂量下对小鼠骨髓细胞染色体无致畸变作用。结论浮海石无致突变作用,用于临床是安全的。     

Wentilactone A的遗传毒性评价

目的 检测Wentilactone A的遗传毒性。方法 应用经典遗传毒性检测组合(Ames试验、体外培养CHO细胞染色体畸变试验和小鼠骨髓微核试验)检测Wentilactone A的遗传毒性。结果 Ames试验结果提示,Wentilactone A在每皿5 000、500、50、5、0.5 μg 5个剂量下,在加和不加代谢活化系统（S9）时,对鼠伤寒沙门菌均无致突变性。CHO细胞染色体畸变试验结果提示,在终浓度23.74、47.48、94.96 μg/ml 3个剂量组,在加和不加S9中,于作用4 h和24 h的条件下培养的CHO细胞,均未诱发染色体畸变。小鼠骨髓微核试验在100、200、400 mg/kg 3个剂量下作用24 h以及400 mg/kg剂量下作用48 h对骨髓细胞的微核诱发率,与溶剂对照组比较均无显著差异（P>0.05）。结论 Wentilactone A对鼠伤寒沙门菌无致突变性,对CHO细胞的染色体无致畸变作用,对ICR小鼠无诱发骨髓细胞微核的效应。上述结果提示Wentilactone A不具有遗传毒性和潜在致癌性。     

3D打印多孔钽浸提液体外细胞毒性和遗传毒性研究

目的 考察3D打印多孔钽浸提液的细胞毒性和遗传毒性.方法 参照GB/T 16886.12标准制备3D打印多孔钽浸提液,采用噻唑蓝(MTT)法检测样品浸提液的细胞毒性,分别采用体外哺乳动物细胞染色体畸变试验、细菌回复突变试验(Ames试验)、体内哺乳动物骨髓细胞微核试验检测样品浸提液的遗传毒性.结果 样品浸提液作用于小鼠...     

红景天苷注射液遗传毒性的研究

目的：检测红景天苷注射液的遗传毒性。方法：应用微生物回复突变实验（Ames实验,5 000、500、50、5.0、0.5μg/皿）、体外培养CHO细胞染色体畸变实验（2 0001、000、500μg/mL）和小鼠骨髓微核实验法（1 500、750、375μg/kg）检测红景天苷注射液的遗传毒性。结果：红景天苷注射液对鼠伤寒沙门菌无致突变性,对体外培养CHO细胞染色体无致畸变作用,对ICR小鼠无诱发骨髓嗜多染红细胞微核的效应,三个实验结果均呈阴性。结论：红景天苷注射液不具有遗传毒性。     

银参胶囊遗传毒性研究

目的探讨银参胶囊的遗传毒性。方法选用SPF级健康ICR小鼠,通过Ames试验、小鼠骨髓细胞微核试验、小鼠睾丸染色体畸变试验等遗传毒性试验验证银参胶囊的安全性。结果 Ames试验中,银参胶囊在8~5 000μg/皿剂量范围内,无论是否加入哺乳动物肝脏微粒体酶(S9),鼠伤寒沙门菌TA97,TA98,TA100,TA102等4株菌的回复突变菌落数均未出现剂量依赖性增加;微核试验中,2 500,5 000,10 000 mg/kg剂量组均未见骨髓中含微核的嗜多染红细胞数增加;小鼠睾丸染色体畸变试验中,药物质量分数为2 500,5 000,10 000 mg/kg时,细胞的染色体畸变率均未出现剂量依赖性增加。结论银参胶囊未显示致突变作用,可初步判定其在遗传毒性方面是安全的。     

双层人工皮肤的生物相容性研究

目的 考察一种新型的皮肤替代物双层人工皮肤的生物相容性.方法 依据GB/T16886医疗器械生物学评价标准规定的细胞毒性、急性毒性试验、皮内刺激试验、致敏试验、亚慢性毒性试验、植入试验和遗传毒性试验方法,评价双层人工皮肤的生物相容性,以验证其安全性和有效性.结果 显示双层人工皮肤无急性毒性、无皮内刺激作用、无致敏作用,细胞毒性不超过1级;皮下植入后1周、4周和12周后,植入组织样品的炎症反应和纤维囊形成与对照无明显差别;遗传毒性采用鼠伤寒沙门氏菌回复突变试验（Ames试验）、体外哺乳动物细胞染色体畸变试验、哺乳动物细胞体外基因突变试验,结果均为阴性,说明双层人工皮肤无遗传毒性.结论 双层人工皮肤具有良好的生物相容性,没有观察到不良反应,符合临床使用要求.     

Evaluating the mutagenic potential of chemicals the minimal battery and extrapolation problems

During the past ten years growing concern about damage to DNA as an important cause of human ill-health has resulted in an explosive development of the field of genetic toxicology. Adequate regulations to restrict exposure to chemical mutagens require recognition and evaluation of mutagenic activity. For this purpose a qualitative and an extrapolation phase can be distinguished. For the qualitative phase, the minimal battery should consist of at least three tests, that is: (1) tests for gene- or point mutations in bacteria (Salmonella or E. coli) with and without metabolic activation; (2) two tests for point mutations in eukaryotes, or (3) one such test and a test for the detection of chromosome aberrations in mammalian cells in vitro. Depending on experience and facilities, a choice of two can be made out of the following four test systems: (1) Tests for point mutations in mammalian cells in vitro, with and without metabolic activation (deficiency for HGPRT, or TK); (2) the sex-linked recessive lethal test with Drosophila melanogaster; (3) tests with yeast, Saccharomyces cerevisiae, for point mutations, with and without metabolic activation; (4) tests for chromosome aberrations in mammalian cells in vitro, with and without metabolic activation. Two different metabolic activation systems should be employed. For further selection of more sensitive test systems, studies on comparative mutagenesis are considered important. A mammalian test for chromosome aberrations in vivo is not included in this minimal battery. Since under in vivo conditions considerably lower concentrations have to be employed than in vitro, it seems unlikely that positive results will be obtained with an in vivo mammalian cytogenetic assay, following negative results in an in vitro cytogenetic assay or in two different tests for point mutations. The finding that the effective concentration for the production of chromosome breakage events differs from that required to induce point mutations (the two-level effect) will be briefly discussed. When mutagenic compounds are indispensible or, in the case of ubiquitous exposure, a quantification of risks becomes necessary and here one is confronted with many difficulties. Information on damage that is hard to measure directly can be obtained in an indirect way by comparison with end-points that can be determined experimentally, such as alkylation per nucleotide. Names of chemical substances tested: hydroxylamine; diepoxybutane; N-ethyl-N-nitrosourea; methylmethanesulfonate (MMS); DEN; Mitomycin C; Procarbazine; atrazine; benz(a)pyrene; EMS; pyrolitic products; flavonoids; mycotoxins; nitrosamines; TEMGiven at the International Conference Mutagenicity Testing of Pharmaceuticals: Present Status, Paris, 12–14 March, 1980, sponsored by the Fondation de l'Industrie Pharmaceutique pour la Recherche     

Appropriate in vitro test conditions for genotoxicity testing of fibers

With the exception of asbestos fibers, little information is available on genotoxicity testing of fibers (i.e., respirable-sized, fiber-shaped particulates, RFP). In contrast to standard genotoxicity testing of soluble substances, fibers bring about specific problems. Test results can be influenced by fiber dimensions, surface properties, and biopersistence. The mechanisms of fiber-induced genotoxicity are not yet clear, but direct interaction with the genetic material and indirect effects via production of reactive oxygen species (ROS) have been proposed. Asbestos did not significantly induce gene mutations in bacterial and mammalian systems but led to a clear induction of structural and numerical chromosome aberrations in cultured mammalian cells. It is the purpose of this article to critically review positive in vitro genotoxicity data obtained for asbestos in the comet assay, the chromosome aberration test, and the micronucleus test and to identify the test conditions that are specifically required for the detection of asbestos-induced genotoxicity. It is concluded that appropriate cell systems are available for testing fiber-induced genotoxicity. Fiber samples have to be well characterized and phagocytosis and cytotoxic effects have to be determined for the correct interpretation of genotoxicity test results. A combination of the micronucleus test and the comet assay using continuous treatment (without exogenous metabolic activation) seems to be well suited to detect genotoxic activity of asbestos fibers with high accuracy. Further investigations are needed to shown whether this approach can be recommended for genotoxicity testing of fibers in general.     

APPROPRIATE IN VITRO TEST CONDITIONS FOR GENOTOXICITY TESTING OF FIBERS

With the exception of asbestos fibers, little information is available on genotoxicity testing of fibers (i.e., respirable-sized, fiber-shaped particulates, RFP). In contrast to standard genotoxicity testing of soluble substances, fibers bring about specific problems. Test results can be influenced by fiber dimensions, surface properties, and biopersistence. The mechanisms of fiber-induced genotoxicity are not yet clear, but direct interaction with the genetic material and indirect effects via production of reactive oxygen species (ROS) have been proposed. Asbestos did not significantly induce gene mutations in bacterial and mammalian systems but led to a clear induction of structural and numerical chromosome aberrations in cultured mammalian cells. It is the purpose of this article to critically review positive in vitro genotoxicity data obtained for asbestos in the comet assay, the chromosome aberration test, and the micronucleus test and to identify the test conditions that are specifically required for the detection of asbestos-induced genotoxicity. It is concluded that appropriate cell systems are available for testing fiber-induced genotoxicity. Fiber samples have to be well characterized and phagocytosis and cytotoxic effects have to be determined for the correct interpretation of genotoxicity test results. A combination of the micronucleus test and the comet assay using continuous treatment (without exogenous metabolic activation) seems to be well suited to detect genotoxic activity of asbestos fibers with high accuracy. Further investigations are needed to shown whether this approach can be recommended for genotoxicity testing of fibers in general.     

Failure of the standard battery of short-term tests in detecting some rodent and human genotoxic carcinogens

Theoretical reasons and experimental evidence indicate that a no-effect level generally cannot be expected for genotoxic carcinogens; as a consequence, in quantitative risk assessment the capability of distinguishing genotoxic from non-genotoxic carcinogens is of fundamental importance in order to identify relevant levels of human exposure. According to generally accepted guidelines, the standard three-test battery for the detection of genotoxic compounds consists of: (i) an in vitro test for gene mutation in bacteria; (ii) an in vitro test in mammalian cells with cytogenetic evaluation of chromosomal damage and/or a test that detects gene mutations; (iii) an in vivo test for chromosomal damage using rodent hematopoietic cells. This test battery is designed to avoid the risk of false negative results for compounds with genotoxic potential, but it cannot be taken for granted that the risk is completely eliminated. As a matter of fact there are some chemicals, classified by the International Agency for Research on Cancer (IARC) as probably or possibly carcinogenic to humans, which gave consistent negative results in this test battery, and in contrast provided positive results in other not routinely employed genotoxicity assays. The failure of the standard test battery in detecting some genotoxic carcinogens is attributable to several causes, but the principal of them are the following ones: in vitro, the artificial metabolic activity of the liver S9-mix, and the different biotransformation of chemicals in cells of different type and from different animal species; in vivo, the pharmacokinetic behaviour of the test compound, and its possible species-, sex- and tissue-specificity.     

Thiourea induces DNA repair synthesis in primary rat hepatocyte cultures and gene mutations in V79 Chinese hamster cells

Thiourea was investigated for its capacity to cause DNA alterations in cultured mammalian cells. The induction of DNA repair in primary rat hepatocyte cultures and of gene mutations in V79 Chinese hamster cells were used as biological endpoints. In hepatocytes, thiourea elicited a linear increase in DNA repair replication in the concentration range tested (5–25 mM). In V79 cells, thiourea (10–40 mM) significantly increased the frequency of 8-azaguanine-resistant mutants. The present results show that thiourea is weakly, but definitely, genotoxic and mutagenic in cultured mammalian cells.     

伊拉地平单室渗透泵控释片的制备及在犬体内的生物等效性研究

目的：制备伊拉地平单室渗透泵控释片,考察其在Beagle犬体内的药动学特征。方法：以伊拉地平为模型药物制备单室渗透泵控释片,并考察其体外释放度。以国外市售伊拉地平控释片为参比制剂,选用8只Beagle犬,采用双周期交叉给药灌服参比制剂或受试制剂（自制伊拉地平单室渗透泵控释片）5mg,给药后O．5、1．0、1．5、2．0、3．O、4．0、5．0、6．0、8．0、12．0、24．0、36．0、48．0、60．0h取前肢静脉血2．0ml,采用超高效液相色谱．串联质谱法测定伊拉地平的血药浓度,以DAS3．2．2计算药动学参数。分析生物等效性。结果：成功制备了伊拉地平单室渗透泵控释片,其体外可持续释放24h。受试制剂和参比制剂的药动学参数t1/2分别为（5．2±2．6）、（6．1±2．5）h,Cmax分别为（8．40±4．25）、（8．47±4．54）ng／ml,AUC0-∞分别为（92．1±53．4）、（96．2±56．3）ng·h／m1,其中Cmax和AUC0-∞的90％可信区间均在80％～125％内,受试制剂对参比制剂的相对生物利用度为95．74％。结论：制备的伊拉地平单室渗透泵控释片实现了24h释药,具有缓释效果,且与参比制剂生物等效。     

美他多辛作用的研究

目的：因美他多辛对乙醇性肝病的生化杌理有积极作用,同时用于急性和慢性乙醇中毒、乙醇性肝病,但该药的致突变作用未见报道。本文在于检测关他多辛致突变性。方法：通过微生物回复突变试验、小鼠微核试验、哺乳动物培养细胞染色体试验证明。结果：美他多辛的三个试验结果均呈阴性。结论：美他多辛在用于治疗急性和慢性乙醇中毒、乙醇性肝病无致突变性。     

Mutagenic evaluation of the organophosphorus insecticides methyl parathion and triazophos in Drosophila melanogaster

The possible genotoxic effects of the organophosphorus insecticides methyl parathion and triazophos were evaluated by their ability to induce gene and chromosome mutations in male germ cells of Drosophila melanogaster. Sex-linked recessive lethal (SLRL), total and partial sex-chromosome losses (SCL), and non-disjunction (ND) assays were conducted. The routes of administration included adult feeding, injection, and larval feeding. Methyl parathion was unable to induce point mutations or chromosome mutations, although a small increase in the frequency of non-disjunction was detected after larval treatment. Triazophos induced point mutations when assayed in the SLRL test and induced a weak increase in the non-disjunction frequency, but gave negative results in the SCL test.     

Particokinetics in vitro: dosimetry considerations for in vitro nanoparticle toxicity assessments.

The rapid growth in the use of in vitro methods for nanoparticle toxicity assessment has proceeded with limited consideration of the unique kinetics of these materials in solution. Particles in general and nanoparticles specifically, diffuse, settle, and agglomerate in cell culture media as a function of systemic and particle properties: media density and viscosity and particle size, shape, charge and density, for example. Cellular dose then is also a function of these factors as they determine the rate of transport of nanoparticles to cells in culture. Here we develop and apply the principles of dosimetry in vitro and outline an approach for simulation of nanoparticle particokinetics in cell culture systems. We illustrate that where equal mass concentrations (mug/ml) imply equal doses for dissimilar materials, the corresponding particle number or surface area concentration doses differ by orders of magnitude. More importantly, when rates of diffusional and gravitational particle delivery are accounted for, trends and magnitude of the cellular dose as a function of particle size and density differ significantly from those implied by "concentration" doses. For example, 15-nm silver nanoparticles appear approximately 4000 times more potent than micron-sized cadmium oxide particles on a cm(2)/ml media basis, but are only approximately 50 times more potent when differences in delivery to adherent cells are considered. We conclude that simple surrogates of dose can cause significant misinterpretation of response and uptake data for nanoparticles in vitro. Incorporating particokinetics and principles of dosimetry would significantly improve the basis for nanoparticle toxicity assessment, increasing the predictive power and scalability of such assays.     

The genotoxicity of diaveridine and trimethoprim

We examined the genotoxicity of diaveridine and trimethoprim in the bacterial umu test, the bacterial reverse mutation test, the in vitro chromosome aberration test, the in vivo rodent bone marrow micronucleus test in two species, and the in vivo comet assay in five mouse organs. Both compounds were negative in the umu test (Salmonella typhimurium TA1535/pSK1002) and in the reverse mutation tests (S. typhimurium TA100, TA98, TA97, TA102, and Escherichia coli WP2 uvrA/pKM101) in the presence and absence of S9 mix. Diaveridine induced structural chromosome aberrations in cultured Chinese hamster CHL cells in the absence of a metabolic activation system, but not in the presence of a liver S9 fraction. No clastogenic activity in CHL cells was detected for trimethoprim. Bone marrow micronucleus tests in mice and rats conducted on diaveridine by single- and triple-oral dosing protocols were negative. The comet assay revealed that a single oral administration of diaveridine significantly induced DNA damage in liver, kidney, lung, and spleen cells, but not in bone marrow cells. The significant increase in migration values of DNA was reproducible with dose-response relationship. We suggest that the liver detoxifies the compound before it reaches the bone marrow, and that is why it is negative in the in vivo bone marrow micronucleus test. We concluded that diaveridine is genotoxic to mammalian cells in vitro and in vivo.