雷公藤内酯醇的遗传毒性评价

目的 观察雷公藤的主要有效成分之一雷公藤内酯醇的遗传毒性。方法 采用鼠伤寒沙门氏菌回复突变试验(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雷公藤内酯醇的遗传毒性。结果 Ames试验提示在每皿1.6~1000 μg受试剂量下,在加或不加S9代谢活化系统时,受试物对组氨酸缺陷型鼠伤寒沙门氏菌TA97、TA98、TA100、TA102及TA1535所诱发的回复突变菌落数均与溶媒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0.01、0.02和0.04 μg/ml 3个剂量的受试物,对加与不加S9代谢活化系统培养的CHO细胞的染色体畸变率无明显影响。小鼠骨髓微核试验设180、360、720 μg/kg 3个剂量,在720 μg/kg剂量时,雷公藤内酯醇有诱发骨髓嗜多染红细胞微核率增高的效应。结论 在本实验条件下,雷公藤内酯醇对鼠伤寒沙门氏菌无致突变性,对哺乳动物培养细胞染色体无致畸变作用,720 μg/kg剂量下对ICR小鼠有诱发骨髓嗜多染红细胞微核率增高的效应。提示雷公藤内酯醇对人体可能具有潜在的遗传毒性。     

虾青素软胶囊遗传毒性研究

目的 研究虾青素软胶囊的遗传毒性.方法 采用鼠伤寒沙门氏茵回复突变试验(Ames试验)、小鼠骨髓细胞微核试验和小鼠精子畸形试验等实验方法对虾青素软胶囊的遗传毒性进行研究.结果 虾青素软胶囊鼠伤寒沙门氏茵回复突变试验受试物各剂量组回变茵落数均未超过自发回变组回变茵落数的2倍,结果为阴性;小鼠骨髓细胞微核试验和小鼠精子畸形试验受试物各剂量组与空白对照组比较,微核率及精子畸变率无明显差异(P＞0.05),实验结果为阴性.结论 在本实验条件下,虾青素无遗传毒性.     

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。     

崖柏精油致突变作用研究

摘 要 目的：采用鼠伤寒沙门菌回复突变（AMES）试验和哺乳动物微核试验对崖柏精油的遗传毒性进行评价。方法: AMES试验采用TA97、TA98、TA100和TA102四种菌株,对崖柏精油的致突变性进行评价。采用小鼠骨髓噬多染红细胞微核试验,对本品的染色体毒性进行评价。结果: 崖柏精油不同剂量组对TA97、TA98、TA100、TA102四种试验菌株,在加大鼠肝微粒体酶S9和不加S9的情况下,回复突变数均不超过溶剂对照组的1倍,差异无统计学意义（P<0.05）,为诱变阴性。微核试验表明,崖柏精油各剂量组对小鼠骨髓微核率无显著影响（P>0.05）。结论:崖柏精油无明显遗传毒性。     

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。     

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。     

灵丹菌质的3次遗传毒性实验研究

目的研究灵丹菌质的遗传毒性,为其开发利用提供依据。方法利用Ames试验、小鼠精子畸形试验以及骨髓细胞微核试验评价灵丹菌质的遗传毒性。Ames试验设立8、40、200、1 000和5 000μg/皿5个剂量组,计算各剂量组在加与不加体外代谢活化系统S9的情况下,对鼠伤寒沙门氏菌TA97、TA98、TA100和TA102的诱发回变菌落数。小鼠精子畸形试验设5、10和20 ml/kg·BW 3个剂量组,计算各剂量组小鼠精子畸形率。小鼠骨髓细胞微核设5、10和20 ml/kg·BW 3个剂量组计算各剂量组微核发生率。结果受试物各剂量组4种菌株的回变菌落数均未超过未处理对照组的2倍;受试物各剂量组的精子畸形率和微核细胞率均未见统计学意义。结论在本研究条件下,灵丹菌质未见遗传毒性。     

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。     

三七护肝胶囊的遗传毒性评价

摘 要 目的：对三七护肝胶囊的急性毒性和遗传毒性进行研究,评价其毒理学安全性。方法: 对三七护肝胶囊进行小鼠经口急性毒性试验、Ames试验、小鼠骨髓细胞微核试验、小鼠精子畸形试验。结果: 三七护肝胶囊对小鼠的急性毒性最大耐受剂量（MTD）大于20 g·kg^-1,属无毒物质;各剂量组Ames试验、微核试验和精子畸形试验结果均为阴性。结论: 在本试验条件下,三七护肝胶囊属于无毒级物质,未显示有遗传毒性作用。     

金锦香提取物毒理学安全性评价

目的研究金锦香提取物的毒理学安全性。方法采用小鼠急性毒性试验、遗传毒性试验、90 d亚慢性毒性试验对金锦香提取物的体内安全性进行评价。结果小鼠急性毒性试验表明,金锦香提取物的小鼠最大耐受剂量(MTD)> 16.0 g·kg~(-1);遗传毒性试验显示,金锦香提取物对鼠伤寒沙门氏菌菌株TA97、TA98、TA100、TA102均未呈现遗传毒性,对小鼠骨髓嗜多染红细胞微核率及嗜多染红细胞与正染红细胞(PCE/NCE)的比值均无影响,对小鼠精子畸形发生无显著作用(P <0.05);90 d亚慢性毒性试验表明,金锦香提取物各剂量组与对照组相比,动物的体重变化、进食量、食物利用率、主要脏器重量、血液学和血液生化指标等均无显著性差异(P <0.05)。结论金锦香提取物在20.00g·kg~(-1)剂量内是安全的,为金锦香的进一步开发利用提供了理论依据。     

Quantitative assessment of a prognostic or predictive biomarker panel

ABSTRACT

Methods for assessing whether a single biomarker is prognostic or predictive in the context of a control and experimental treatment are well known. With a panel of biomarkers, each component biomarker potentially measuring sensitivity to a different drug, it is not obvious how to extend these methods. We consider two situations, which lead to different ways of defining whether a biomarker panel is prognostic or predictive. In one, there are multiple experimental targeted treatments, each with an associated biomarker assay of the relevant target in the panel, along with a control treatment; the extension of the single-biomarker scenario to this situation is straightforward. In the other situation, there are many (nontargeted) treatments and a single assay that can be used to assess the sensitivity of the patient’s tumor to the different treatments. In addition to evaluating previous approaches to this situation, we propose using regression models with varying assumptions to assess such panel biomarkers. Missing biomarker data can be problematic with the regression models, and, after demonstrating that a multiple imputation procedure does not work, we suggest a modified regression model that can accommodate some forms of missing data. We also address the notions of qualitative interactions in the biomarker panel setting.     

The effect of dimethylaminoadamantane on neuronal membranes.

The effect of dimethylaminoadamantane (DMAA), an amantadine derivative with an anti-Parkinson property, on rat sensory nerve fibres was studied with the sucrose gap method. DMAA 10(-4) M in normal Locke solution reduced the spike amplitude without changing the resting potential, increased the membrane resistance and depressed repetitive spike activity elicited by depolarizing currents. From experiments performed with changed concentrations of sodium, potassium, calcium and chloride ions in the suspension medium it appears that the permeability of sodium, potassium and chloride ions is reduced by DMAA. The possible implication of the membrane effects of the drug in its action on dopaminergic transmission in the brain is discussed.     

In vitro and in vivo genotoxic evaluation of Bothrops moojeni snake venom

Abstract

Context: Bothrops moojeni Hoge (Viperidae) venom is a complex mixture of compounds with therapeutic potential that has been included in the research and development of new drugs. Along with the biological activity, the pharmaceutical applicability of this venom depends on its toxicological profile.

Objective: This study evaluates the cytotoxicity and genotoxicity of the Bothrops moojeni venom (BMV).

Material and methods: The in vitro cytotoxicity and genotoxicity of a pooled sample of BMV was assessed by the MTT and Comet assay, respectively. Genotoxicity was also evaluated in vivo through the micronucleus assay.

Results: BMV displayed a 50% cytotoxic concentration (CC₅₀) on Vero cells of 4.09?µg/mL. Vero cells treated with 4?µg/mL for 90?min and 6?h presented significant (p?<?0.05, ANOVA/Newman–Keuls test) higher DNA damage than the negative control in the Comet assay. The lower DNA damage found after 6?h compared with the 90?min treatment suggests a DNA repair effect. Mice intraperitoneally treated with BMV at 10, 30, or 80?µg/animal presented significant genotoxicity (p?<?0.05, ANOVA/Newman–Keuls test) in relation to the negative control after 24?h of treatment. Contrary to the in vitro results, no DNA repair seemed to occur in vivo up to 96?h post-venom inoculation at a dose of 30?µg/animal.

Discussion and conclusion: The results show that BMV presents cyto- and genotoxicity depending on the concentration/dose used. These findings emphasize the importance of toxicological studies, including assessment of genotoxicity, in the biological activity research of BMV and/or in the development of BMV-derived products.     

The human tumor colony-forming chemosensitivity assay: A biological and clinical review

Summary Over forty papers describing correlations between in vitro human tumor sensitivity to a variety of chemotherapeutic agents and the in vivo response of patients to those agents have been published since the publication in 1978 by Salmon and Hamburger of their results of a human tumor colony-forming chemosensitivityassay (CFCA). The true positive rate in over 1600 correlations is 71% and the true negative rate is 94%. The biological elements of the assay, its developmental history, its place in the spectrum of in vitro chemosensitivity assays, and its theoretical and practical limitations are discussed. The scope, design, and limitations of key clinical trials are presented and an analysis of the potential errors of statistical interpretation of the trials as well as the results of the trials is given.     

Genotoxicity studies on HM10760A,recombinant human erythropoietin conjugated to globin fragment

HM10760A is a recombinant human erythropoietin chemically conjugated to the N-terminus of human immunoglobulin Fc fragment through a polyethylene glycol linker. HM10760A was shown to have a relatively long half-life, compared with unconjugated recombinant erythropoietin. In this study, the genotoxicity of HM10760A was investigated by using a test battery of three different methods. In the Ames assay, five strains (TA100, TA1535, TA98, TA1537, and Escherichia coli WP2 uvrA) were tested at six concentrations of 3.13, 6.25, 12.5, 25, 50, and 100?μg/plate. HM10760A did not increase the number of revertant colonies in any tester strains with and without metabolic activation by rat-liver S9 mix. Subsequently, in vitro chromosomal aberration test, using Chinese hamster lung cells, were conducted at the concentrations of 25, 50, and 100?μg/mL. HM10760A did not induce chromosomal aberrations either in the short-period (6 hours) test with or without rat-liver S9 mix or in the continuous-treatment (24 hours) test. In the in vivo bone marrow micronucleus assay using the male ICR (imprinting control region) mouse, HM10760A was subcutaneously administered twice at 24-hour intervals at doses of 0, 150, 300, and 600?μg/kg. HM10760A produced a slight, but statistically significant, increase in the frequency of micronucleated polychromatic erythrocytes at 600?μg/kg. However, no biological significance was assumed, because this value was within the historical control range. From these findings obtained from the genotoxicity assays performed in this study, it appears unlikely that HM10760A acts as a genotoxic agent in vitro and in vivo.     

终止分光光度法快速检测酪氨酸多巴氧化酶活性

目的 提高分光光度法检测酪氨酸多巴氧化酶活性(多巴氧化酶)的灵敏度、特异性和稳定性.方法 用高氯酸对分光光度法进行改进,并与多巴色素法和连续检测法比较.结果 终止分光光度法检测多巴氧化酶的灵敏度为多巴色素法10倍,为连续监测法的2倍,且稳定性强,可分析浊样和非浊样样本,并且每小时样本测定量为另两种的2倍.结论 建立的终止分光光度法检测多巴氧化酶活性敏感、快速和特异,是一种可靠的实验诊断手段.     

HPLC法测定交沙霉素的含量及与微生物检定法的比较

目的:建立HPLC法测定交沙霉素含量的方法并与微生物检定法结果相比较.方法:采用RP-HPLC法,以C18为固定相;流动相:水-乙腈(45:55),用三乙胺调节pH值为8.0;检测波长:232 nl.结果:在0.1～1.4 mg·ml-1的浓度范围内呈良好线性关系.结论:本方法色谱系统简单,可靠.经与微生物检定法结果比较,结果准确可靠,精密度高.且能将交沙霉素与其各杂质组分实现良好分离,测定结果更加准确.     

A review of in vitro methods to assess the biological activity of tobacco smoke with the aim of reducing the toxicity of smoke

In the last few years tobacco companies have been developing several research strategies in order to reduce the risks associated with smoking. These strategies include, for example, the refining of alternative cigarette designs that reduce the amount of hazardous chemicals in the mainstream smoke by introducing modified filters, and/or reducing the amount of biologically significant ingredients in tobacco-burning cigarettes. In the last few decades numerous studies have been published to assess the biological activity of tobacco smoke using in vivo and in vitro test systems. In this scenario a general scientific consensus on how to measure and characterize the risk associated with cigarette smoke is still lacking. Short-term in vitro assays, which are widely accepted by regulatory agencies around the world, are useful tools to evaluate both the biological activity and the progress towards a reduction of tobacco smoke toxicity. These assays could be mainly applied to evaluate cytotoxicity and genotoxicity properties on whole cigarette smoke as well as condensates or fractions of whole smoke. Cytotoxicity induction can be measured as cellular viability and growth rates using different end-points. Otherwise, the target of genotoxicity studies is the DNA molecule. For genotoxicity evaluation, the end-points and cell systems should be chosen from those that are relevant and appropriate as clinical surrogate markers. In this respect, the occurrence of early biological effect markers, such as mutational or clastogenic events (point mutations, frameshifts, micronuclei, SCE, DNA adducts) in bacterial and mammalian cells should be studied in a tiered approach following the guidelines of regulatory agencies. The choice of criteria shall be matter of discussion.     

自动加样快速微孔板法在疑难配血中的应用

目的 在疑难配血中建立一种自动化、快速、准确的供血者标本血型抗原筛查方法.方法 使用自动加样快速微孔板法对188个供血者全血标本进行C、e、M、Fya、Jka抗原检测,同时使用手工试管法进行平行实验;记录并对比上述两种方法筛查总过程使用的时间,筛查出的供者标本数.用微板法和试管法对抗-C、抗-e、抗-M、抗-Fya、抗-Jka试剂进行抗体效价测定.结果 微孔板法、试管法均筛查出相同血型抗原阴性供者,微孔板筛查所需要的总时间较短.效价测定显示微板法灵敏度与手工试管法相同.结论 微孔板法在疑难配血中筛查特定抗原阴性的供者,具有快速、准确的效果.     

MTT法检测rhEGF生物活性

目的鉴于待测的生物活性因子均有适于其自身的最佳测定条件,规范了重组表皮生长因子(rhEGF)的MTT测定法。方法根据MTT测定法原理,在3个因素(每孔细胞数、MTT和胎牛血清浓度)及3个水平上进行了正交实验设计。结果MTT检测法的最佳条件是:Balb/c3T3细胞悬液(5×107cells·L-1)100μl·well-1,MTT(8g·L-1)20μl·well-1,FBS(1·5%)100μl·well-1。结论上述正交实验规范的MTT最佳实验条件,适用于Balb/c3T3细胞活性的检测及其药物抑制性实验。但在测定rhEGF生物活性时,为使对照孔细胞数保持在较低水平以便给生物活性因子留有适当的余地以显示其促细胞增殖活性,建议FBS浓度采用0·5%。