胚胎移植法在生殖发育毒理学中的应用

随着环境致畸物种类、数量的增多，使生殖毒性检测技术迅速发展起来。在研究内容上，由单纯致畸实验发展到其他雌、雄性生殖毒性实验；在研究手段上，由整体动物实验扩大至离体器官、细胞培养；在研究水平上由细胞水平发展到分子水平。胚胎移植技术的建立和成熟使生殖毒理学检测技术又上了一个新的台阶。     

女（雌）性生殖毒性的研究策略和方法

女（雌）性生殖毒性的研究策略和方法吴向东１丁训诚１综述蒋学之２审校１．中国生育调节药物毒理检测中心（上海２０００３２）２．上海医科大学公共卫生学院近年来，大量的化学物不断进入环境和人们的日常生活，这些外源性化学物对女（雌）性生殖系统的影响越来越受重视...     

湖北省药物安全性评价中心成立

20 0 4年 9月 13日 ,湖北省科技厅批准依托湖北省医药工业研究院有限公司成立“湖北省药物安全性评价中心”(简称中心 )。该中心由一般毒理室、局部毒理室、遗传及生殖毒理室、安全性药理室、病理室、临床检验室、动物实验室等组成 ,拥有国内一流的实验检测仪器。其人员结构相对合理、仪器设备比较齐全、动物实验设施较为完备、管理体系比较完善 ,并严格按照药品非临床实验管理规范 (GLP)要求运行。已开展了单次和多次给药毒性试验、生殖和遗传毒性试验、局部毒性试验、免疫原性试验及安全性药理等药物非临床安全性评价研究。该中心将立足…     

茶叶抗氧化剂对大鼠繁殖和喂养致畸试验

我们在进行了茶叶抗氧化剂对大鼠90天喂养试验的基础上又观察了对大鼠的繁殖和喂养致畸。剂量分组:对照,1000,5000,10000PPm。大鼠喂养13周后,以查到精子之日作为怀孕0天。受精鼠一半作繁殖试验,(记录孕鼠分娩情况,新生仔鼠有无外观畸形,生产当天,第四天,第21天称仔鼠体重并记录活仔鼠数)。一半作喂养致畸试验,(剖     

生殖毒性研究进展

随着毒理学新理论和新技术的不断涌现,生殖毒性的研究也随之不断深入;《药物生殖毒性研究技术指导原则》(2006年11月)的出台[1],促进了我国药物生殖毒性的研究水平迈入新高。结合我们多年来药物生殖毒性研究的经验和体会,现以药物生殖毒性研究为主线,对生殖毒性研究进展总结并归纳如下:1药物生殖毒理研究范畴更加深入细致1.1生殖器官概念的扩大化传统生殖毒性的研究中的雄性性器官,包括生殖器官和附属腺体,主要指的是睾丸、附睾、输精管、射精管和阴茎等[2-3],而近年来的概念逐步扩展到与生殖相关的腺体和器官,如前列腺、包皮腺、精液囊、提…     

我国遗传毒理学发展的回顾

回顾了中国遗传毒理学发展的三个阶段 :自 70年代末至 1983年为启动阶段 ,建立了一系列遗传毒性检测方法 ,筛检了大量环境化学物的遗传毒性 ,以及培训了大量专业人员 ,成立了中国环境诱变剂学会。自 1983～ 1993年为深入发展的关键阶段。遗传毒性检测方法标准化、规范化 ,开展遗传毒性机制的研究 ,遗传毒性试验列入新药、农药、食品、化妆品等安全性评价准则 ,并成立了中国毒理学会遗传毒理专业委员会。自 1993年至今 ,遗传毒理学进入了分子时代。建立了分子致突变测试系统 ,例如穿梭质粒载体系统 ,转基因动物致突变测试系统 ,以及其他应用分子生物学方法进行突变的分子分析等。开展了基因突变分子机制的研究 ,在非定标性突变方面取得了显著成就。并且 ,还回顾了遗传毒理在我国人类环境的现场监测、人群健康监测、遗传毒性与疾病、肿瘤的流行病学调查中的应用     

化学物质生殖毒性替代方法的研究现状

科技发展和人类进步,促进3R理论发展,即减少、替代和优化试验动物的使用。因此,众多努力用于毒理学替代方法的研究。生殖和发育毒性检测中使用的动物数量最多,但因哺乳动物生殖周期的复杂性,生殖发育毒性替代方法的研究进展缓慢。胚胎干细胞、微团检测和全胚胎培养试验,以及非洲蟾蜍胚胎试验,可作为发育毒性的筛检方法。利用人生殖细胞体外模型进行生殖毒性研究是一个挑战。跨领域技术、传感器技术、(Q)SARs及各种组学技术的迅速发展,都将使生殖发育毒性替代方法的研究得到新的发展和突破。     

第十届全国毒理学术研讨会征文通知

中国药理学会毒理专业委员会决定召开中国药理学会第十届全国毒理学术研讨会。会议承办单位 :重庆第三军医大学药理教研室北京昭衍新药研究中心1.征文内容 :关于药品和各种化学品安全性评价 ;替代性毒理学试验 ;体外毒理学 ;靶器官毒理学 ;遗传毒理和分子毒理学 ;生殖毒理学和发育毒理学等。2 .大会报告内容 :我国药物毒理实验指定原则 ;注册办法中有关毒理学要求 ;生殖毒理学进展 ;遗传毒理学进展 ;靶器官毒理研究 ;替代性毒理试验和体外毒理学研究进展等等。会议将组织有关专题讨论会 :包括GLP筹建和运行的经验和体会 ,遗传毒理学 ,生殖…     

国外已上市药物注册申请中药理毒理文献资料的选择

已在国外上市销售但尚未在国内上市销售,且无知识产权保护的药物长期以来一直是国内新药研发的主要方向。此类药物在国内注册时,药理毒理研究资料中的部分内容通常可以用文献资料予以代替,这些内容一般可包括主要药效学试验、一般药理学试验、急性毒性试验、长期毒性试验、遗传毒性试验、生殖毒性试验和致癌性试验。在此类新药的注册申请过程中,如何选择和提交药理毒理文献资料是一个常见的问题。本文将基于国外已上市药物的特点,从技术评价的角度,对于此类药物药理毒理文献资料的选择和提交提出一些个人观点。1药理毒理文献资料的作用在新…     

橙汁粉末香精毒理学研究及安全性评价

目的研究橙汁粉末香精的毒性和安全性。方法按照《食品安全毒理学评价程序和方法》进行试验。结果橙汁粉末香精对小鼠急性毒性实验LD50=9600mg·kg-1,属实际无毒级;通过Ames试验、骨髓细胞微核试验、小鼠精子畸形试验;传统致畸实验,发现橙汁粉末香精无遗传毒性;30天喂养实验以及90天喂养实验,发现对动物的体重、血液常规及血清生化指标均无明显毒性作用(P>0.05)。结论根据以上结果可基本断定橙汁粉末香精是一种安全、无毒的食品、药品矫味剂。     

Evaluation of OECD screening tests 421 (reproduction/developmental toxicity screening test) and 422 (combined repeated dose toxicity study with the reproduction/developmental toxicity screening test)

The Advisory Committee on Existing Chemicals (BUA) of the Federal Republic of Germany convened a panel with expertise in reproductive and developmental toxicology to evaluate the OECD Screening Tests 421 (Reproduction/Developmental Toxicity Screening Test) and 422 (Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test) with respect to their ability to unmask any potential toxic effects on reproduction. The original assignment for that panel was to "validate" those screening tests. However, the panel members recognized beforehand that this was actually an impossible task because of lack of a sufficient database. Only five chemicals with known reproductive toxicity had been examined following the OECD Screening Test Guidelines 421 or 422. A comparison of these test results with those of the definitive OECD Test Guidelines 414, 415, 416, or additional investigations could, therefore, only have been made with this very limited number of chemicals that had also undergone evaluation by one of the test guidelines cited. In each case biological properties relevant to reproductive toxicity were also indicated by the OECD Screening Tests 421 or 422. This communication reviews the main differences in study design of OECD Screening Test Guidelines 421 and 422 compared to those definitive test guidelines of similar study design for reproduction or developmental toxicity (especially with the one-generation study, OECD Test Guideline 415). The very limited possibilities of detecting late postnatal and postlactational manifestations are emphasized, as is the low statistical power of the OECD Screening Tests 421 and 422. Furthermore, the very limited ability to unmask teratogenicity is delineated. The outcome of screening tests was evaluated based on the results of 57 studies conducted according to the OECD Test Guideline 421 or 422. The test results were categorized according to the incidence of toxic effects on reproduction in the parent animals or their offspring and related to general toxic effects. Based on the ranking of these results, recommendations regarding setting rational priorities for further evaluations of existing chemicals' reproductive hazards are made. In general, the reviewer panel supports the OECD position that the screening tests are useful for initial hazard assessment and can contribute to the decision-making process on setting priorities for further test requirements. The panel also agrees with the OECD statement that the OECD Screening Tests 421 and 422 are neither an alternative to definitive tests (i.e., OECD Test Guidelines 414, 415, and 416) nor are they intended as their replacement.     

透明质酸钠的毒理学研究

目的通过透明质酸钠(SH)的毒理学试验,探讨其食用安全性。方法按GB15193-2003食品安全性毒理学评价程序和方法的规定进行急性毒性试验、三项遗传毒性试验和90 d喂养试验。结果 SH属实际无毒,未见有致突变作用。SH喂养大鼠90 d,动物生长活动正常,未见对动物体重、食物利用率、血常规、血生化、脏器重量及脏体比、病理组织学检查结果有不良影响。结论 SH为无毒物质。     

Freshwater toxicity testing using rehydrated Philodina sp. (Rotifera) as test animals

Rotifers have become widely used in aquatic toxicology as a rapid screening test for toxicity. The commercial availability of diapausing embryos (cysts) have facilitated their popularity because test animals can be obtained without having to master the details of culturing. Other rotifer species have life stages capable of surviving desiccation and also could be used in non‐culture systems for toxicity assessment. In this article, we describe a system for toxicity testing in freshwater based on rehydrating desiccated bdelloid rotifers in the genus Philodina. These animals can remain in this anhydrobiotic state for more than one year and then rehydrate within hours to provide animals for toxicity tests. We describe three endpoints: a 1.5 h ingestion test, a 24 h mortality test, and a five day reproductive test. The latter test requires feeding and a method using a dried commercial product is explained. Using desiccated rotifers and dried food in toxicity tests make this system especially attractive because of its flexibility and low threshold of biological expertise required to execute the tests. The use of the Philodina toxicity test is illustrated with four metals: copper, lead, mercury and cadmium. Reproduction generally was the most sensitive endpoint, with EC50s of 0.33, 0.44, 0.60, and 0.12 mg/L, respectively. Ingestion was a close second with EC50s of 0.13, 1.64, 0.64, and 6.26 mg/L, respectively.     

辅助生殖培养用液中庆大霉素临床前安全评价思考

目的：归纳总结庆大霉素对配子和胚胎的影响,并对辅助生殖培养用液临床前评价需要考虑的问题进行分析,为辅助生殖培养用液产品中庆大霉素的临床前安全评价提供参考。方法：以辅助生殖培养用液中添加的药物庆大霉素为研究对象,结合庆大霉素在产品中的应用,采用文献调研法总结归纳了庆大霉素对生殖细胞、胚胎以及胚胎干细胞产生的影响,并根据辅助生殖用液产品监管现状,提出临床前安全评价思考。结果与结论：庆大霉素被广泛应用在辅助生殖培养用液中,安全、规范地使用庆大霉素在辅助生殖技术中具有重要意义。目前有关庆大霉素对配子及胚胎毒性的研究较少,已知的结论都是基于鼠胚试验及其他动物试验得出的结果扩展到人类胚胎,有必要关注庆大霉素对人类配子及胚胎发育的影响。其次,对于辅助生殖用液产品中庆大霉素添加量的确定、原料质量控制以及检测方法等方面存在的问题提出有针对性的建议,为加强辅助生殖用液的质量控制和产业监管提供思路,从而促进辅助生殖技术的发展。     

Toxicology and safety of anti-oxidant of bamboo leaves. Part 1: Acute and subchronic toxicity studies on anti-oxidant of bamboo leaves.

The anti-oxidant of bamboo leaves (AOB) has recently been certificated as a novel kind of natural anti-oxidant by the Ministry of Health of the People's Republic of China, and has been used in various food systems. Here, AOB was subjected to a series of acute and subchronic toxicological tests to evaluate its safety. It was examined to evaluate acute oral toxicity by using Kun-Ming mice and Sprague-Dawley rats, and its mutagenic potential assessed by reverse mutation test using Salmonella typhimurium, bone marrow cell micronucleus test using Kun-Ming mice, and sperm abnormality test using Kun-Ming mice. In addition, a 90-day oral toxicity study using Sprague-Dawley rats was conducted to evaluate subchronic toxicology. The results showed that the maximum tolerated dose (MTD) of AOB was >10 g/kg body weight in both rats and in mice, which can be regarded as virtually non-toxic. No mutagenicity evidence was detected in any of the three mutagenic tests. Administration at levels of 1.43, 2.87 and 4.30 g/kg per day to the rats for 90 days did not induce significant hematological, clinic, chemical and histopathological changes, and suggested a no-observed-adverse-effect level (NOAEL) of 4.30 g/kg per day. These results indicate that AOB can be generally regarded as safe for use as a food additive.     

三硝基甲苯的遗传毒性研究

用果蝇伴性隐性致死（ＳＬＲＬ）实验和非整倍体检测实验，对三硝基甲苯（ＴＮＴ）的遗传毒性进行了研究。结果表明：雄性果蝇在５４．０ｍｇ、２７．０ｍｇ、１３．５ｍｇ／１００ｍｌ培养基浓度下喂饲２４小时，在ＳＬＲＬ实验中，可引起生殖细胞隐性致死率明显增加，并有明显的剂量反应关系；在非整倍体检测实验中，Ｘ或Ｙ染色体丢失率明显增加，也有明显的剂量反应关系。说明ＴＮＴ对果蝇生殖细胞具有明显的致突变作用。分阶段（２，３，３天）实验表明，ＴＮＴ对果蝇各阶段生殖细胞具有相同的致突变作用。     

Nonclinical safety of ziconotide: an intrathecal analgesic of a new pharmaceutical class

Ziconotide, a potent, selective, reversible blocker of neuronal N-type voltage-sensitive calcium channels, is approved in the United States for the management of severe chronic pain in patients for whom intrathecal therapy is warranted, and who are intolerant or refractory to other treatment, such as systemic analgesics, adjunctive therapies, or intrathecal morphine. In the European Union, ziconotide is indicated for the treatment of severe chronic pain in patients who require intrathecal analgesia. Nonclinical investigations of ziconotide included a comprehensive characterization of its toxicology, incorporating acute and subchronic toxicity studies in rats, dogs, and monkeys; reproductive toxicity assessments in rats and rabbits; and mutagenic, carcinogenic evaluations performed in vivo and in vitro. Additional investigations assessed the potential for cardiotoxicity (rats) and immunogenicity (mice, rats, and guinea pigs), and the presence or absence of intraspinal granuloma formation and local cell proliferation and apoptosis (dogs). The resulting nonclinical toxicology profile was predictive of human adverse events reported in clinical trials and consistent with ziconotide's pharmacological activity. Frequently observed nonclinical behavioral effects included tremoring, shaking, ataxia, and hyperreactivity. Occurrences were generally transient and reversible upon cessation of treatment, and intolerable effects occurred at doses more than 45 times the maximum recommended clinical dose. Ziconotide was not associated with target organ toxicity, teratogenicity, or treatment-related gross or histopathological changes; it displayed no mutagenic or carcinogenic potential and no propensity to induce local cell proliferation or apoptosis. Although guinea pigs developed systemic anaphylaxis, antibodies to ziconotide were not detected in mice, rats, or guinea pigs, indicating low immunogenic potential. No evidence of granuloma formation was observed with intrathecal ziconotide treatment. In summary, the results from these nonclinical safety assessments revealed no significant toxicological risk to humans treated with ziconotide as recommended.     

Validation of whole chick embryo cultures, whole rat embryo cultures and aggregating embryonic brain cell cultures using six pairs of coded compounds

A comparative study was performed to assess the effects of six pairs of coded compounds using cultures of whole chick and rat embryos as well as aggregating brain cell cultures. Developed originally for basic studies in developmental biology, these three culture systems have been adapted for the screening of chemicals in the field of prenatal toxicology. Chick and rat embryos were cultured for 2 days during the early stages of organogenesis. Aggregating cell cultures were prepared from early foetal rat telecephalon and grown for 14 days in a chemically defined medium. Concentration-response relationships were established by treating whole embryos in vitro for 2 days, and aggregating brain cell cultures for 9 days. After decoding the compounds, the results showed that, in the three test systems, specific effects were induced at comparable concentration levels. Similar compound-related malformations could be observed in both chick and rat whole embryo cultures. In aggregating brain cell cultures, neuron- and glia-specific effects could be distinguished. Based on the results obtained in the three in vitro systems, the following concentration ranges were determined for the teratogenic/toxic potencies of the test compounds (in mol/litre): <10⁻⁶: retinoids (Ro 13-6307, Ro 1-5488), 6-aminonicotinamide, ketoconazole; 10⁻⁶−10⁻³: 4-hydroxypyridine, sulfadiazine, sulfanilamide, caffeine, theophylline, metronidazole, methoxyacetic acid; >10⁻³: methoxyethanol. In general, the three in vitro test systems were found to provide concordant and complementary data on the toxicity and teratogenicity of a given compound. These data were also comparable with those available from in vivo studies. It is therefore concluded that such a test battery could contribute significantly to risk assessment and to the reduction of in vivo experimentation in reproductive toxicology.     

TestSmart-high production volume chemicals: an approach to implementing alternatives into regulatory toxicology.

This article examines the status and application of alternatives defined as replacements, refinements, and reduction for screening high production volume (HPV) chemicals. It specifically focuses on the Screening Information Data Set (SIDS), a series of toxicological tests recommended by the Organization for Economic Cooperation and Development to screen such chemicals. Alternative tests associated with acute, repeat-dose, genetic, and reproductive and developmental toxicity were examined at 2 meetings of academic, industry, and regulatory scientists and their status determined. Tests were placed in 1 of 3 categories: ready for immediate use, in need of or currently undergoing validation, or needing research/developmental work. With respect to traditional acute toxicity testing, the basal cytotoxicity approach was placed in the category of research with the up-and-down, fixed-dose, limit test, and the acute toxic class categorized as available for immediate use and the neutral red assay under validation. Cell culture methods that could provide information on acute target organ toxicity were all categorized in the research stage. Studies of the Ah receptor were placed under validation. All alternative tests for repeat-dose toxicity were placed in the category of research. With regard to genetic toxicity, the Ames, mouse lymphoma, and Chinese hamster ovary methods were considered ready for immediate use, while the in vitro micronucleus and Syrian hamster ovary assays were placed in the validation category. All alternatives for developmental toxicity, with the exception of gene chip technology, were placed in the category of validation. Gene chip technology is considered to be in the research stage. For reproductive toxicity, sperm motility and morphology were considered as ready for immediate use, with the other assays categorized as needing validation or in the research stage. Follow-up to these results is obvious. Work needs to be conducted to move those tests from the research stage to the validation and use stage. This is one approach to the development of alternatives to SIDS. Progress along these lines would apply not only to SIDS but also to toxicology in general.     

体外全胚胎培养技术及其在药物生殖发育毒性评价中的应用

体外全胚胎培养（whole embryo culture，WEC）技术是一项在受控的体外环境中培养动物胚胎的技术，是研究药物及化学物质在胚胎发育过程中的药理毒理及作用机制的重要方法之一。本文从大鼠和兔的WEC技术发展概况、培养条件、胚胎发育终点的评分系统和拓展的胚胎发育评价终点、药物生殖发育毒理学应用等方面，对WEC技术及近年来研究进展进行介绍。WEC技术是药物生殖发育毒性研究中重要的替代学研究方法之一。其中，对大鼠与兔组合的WEC研究更具有重要价值，可为胚胎发育毒性研究提供更全面的信息。