紫外线对小白鼠免疫遗传的损伤效应

以ICR雄性小白鼠为实验动物,以紫外线为诱变剂,研究了紫外线对ICR小白鼠的免疫遗传的损伤效应。我们采用了小白鼠骨髓嗜多染红细胞(polychromatic erythrocyte,PCE)微核试验、小白鼠骨髓细胞染色体畸变实验等方法。通过测定小白鼠血象、血红蛋白含量、胸腺及脾指数;小白鼠肝脏过氧化氢酶(catalase,CAT)活性;小白鼠PCE的微核率、染色体畸变率等指标。结果显示,实验照射时间及剂量的紫外线引起小白鼠胸腺指数以及脾指数明显下降,血红蛋白含量和CAT的含量也都明显下降。红细胞数、白细胞数以及血小板数也明显下降。PCE微核率以及骨髓细胞染色体畸变率均明显上升。结果说明紫外线对小白鼠的免疫系统以及遗传物质具有明显的损伤效应。     

氨氮胁迫对黄河鲤幼鱼红细胞微核、核异常的影响

以黄河鲤幼鱼为实验动物,研究氨氮暴露对鲤鱼红细胞微核、核异常的影响.结果表明:氨氮可引起鲤鱼红细胞微核及核异常率发生明显变化,随着氨氮暴露浓度的增加,鲤鱼红细胞微核及核异常率呈现规律性上升变化,且方差分析差异显著(P<0.01);随着氨氮暴露时间的延长,红细胞微核率呈现上升趋势,核异常及总核异常率呈现先升再降趋势,方差分析差异不显著(P>0.05).因此在试验浓度范围内氨氮对鲤鱼红细胞具有明显的遗传毒性,高密度养殖条件下氨氮的毒性不容忽视.     

巯基乙酸对中华蟾蜍红细胞核异常的影响

试验以健康的中华蟾蜍为实验动物,研究注射巯基乙酸对红细胞核异常的影响.试验结果表明,在一定的范围内,巯基乙酸可引起蟾蜍微核细胞率和核异常细胞率遗传指标发生明显变化,随着巯基乙酸浓度的增加和作用时间的延长,蟾蜍红细胞微核和核异常率呈现先上升后下降的规律性变化.     

除草剂丁草胺对蟾蜍红细胞微核及核异常的影响

本文研究除卓剂丁革胺对蟾蜍红细胞核的诱变效应。采用体内红细胞微核测定法,观察蟾蜍在不同浓度和不同的染毒时间红细胞微核及核异常的变化。结果表明,在一定的范围内,丁草胺可引起蟾蜍微核细胞率和核异常细胞率等遗传指标发乍明显变化,随着丁草胺浓度的增加和作用时间的延长,蟾蜍红细胞微核和核异常率呈现先上升后下降的规律性变化。丁草胺的作用具有双向性。一定利量的除草剂丁草胺对蟾蜍红细胞具有明显的遗传毒性,除草剂对水体的污染不容忽视。     

N_1N''''—二甲基甲酰胺对小鼠骨髓多染性红细胞微核出现率的效应

本研究采用微核测定方法来评价N,N′—二甲基甲酰胺(N,N′—Dimethyl formamide简称DMF)对小鼠骨髓多染性红细胞微核出现率的致突变效应。处理结果表明:单次注射不同剂量的DMF,其中1.0mg/kg(体重)剂量组的多染性红细胞微核出现率有明显的上升趋势(4.077‰),当剂量增加到6.0mg/kg时,多染性红细胞微核出现最高峰,微核率为4.374‰,表明该剂量使微核显著增高;随着剂量的继续增加微核出现率反而随之下降。 多次连续注射(每隔24小时注射一次)DMF1.0mg/kg微核出现率为5.204‰,比单次注射相同剂量的微核率高,说明药物有积累的遗传毒理作用。     

阿维菌素对黑斑侧褶蛙毒理效应的研究

本文选用生物农药阿维菌素作为染毒试剂,以农田中常见黑斑侧褶蛙Pelophylax nigromaculatus为研究对象,分别从急性毒性、亚急性毒性、遗传毒性和生理毒性四个方面的实验综合探讨了阿维菌素对黑斑侧褶蛙的生态毒理学效应。急性毒性实验结果表明,阿维菌素对黑斑侧褶蛙24h、48h、72h、96h的LC50分别为:0.3227mg/L、0.0966mg/L、0.0623mg/L、0.0432mg/L,其对黑斑侧褶蛙的安全浓度(SC)为0.004316mg/L。亚急性毒性实验结果表明,暴露期间,随着浓度的升高,毒害作用越大,其对黑斑侧褶蛙肝脏组织损害也越大,而经恢复期后,各实验组肝脏组织受损程度较暴露期都有降低。红细胞核异常及微核诱导实验结果表明,微核和核异常现象与阿维菌素浓度的相关性有一定的范围;随着时间的延长,红细胞核异常率及微核率先上升后下降。肝脏LDH同工酶聚丙烯酰胺凝胶电泳实验结果表明,在一定时间条件下,随浓度的升高,LDH1和LDH2酶带活性呈现降低趋势,LDH4酶带活性无明显变化,LDH5酶带活性先升高后再降低,在同一浓度条件下,随着时间的延长,肝脏LDH同工酶活性均呈升高的趋势。实验结果表明阿维菌素对黑斑侧褶蛙能够造成不同程度的生理、病理及遗传性损伤。     

污染水域鲫鱼外周血细胞形态和数量的变化

为探讨污染环境对鱼类外周血细胞形态及数量的影响,本研究选择相对无污染的刘家峡水库和污染较严重的黄河白银段为研究地点,以鲫鱼(Carassius auratus)为研究对象,常规方法制血涂片,Giemsa染色,统计红细胞微核率、核异常率和各类血细胞数量,并用带有数码采集头(Motic B5 Professional Series)的显微镜拍照各类细胞。结果表明,与刘家峡水库相比,黄河白银段鲫鱼红细胞数量显著减少(P<0·05),核异常率(P<0·05)和微核率(P<0·01)显著增加;白细胞总数和淋巴细胞数量显著增加(P<0·05),而嗜中性粒细胞(P<0·01)和血栓细胞(P<0·05)数量显著减少,单核细胞数量虽有减少趋势但两地差异不显著(P>0·05)。表明黄河白银段污染对鲫鱼外周血细胞数量及红细胞微核率和核异常率具有显著的影响。     

壬基酚对黑斑蛙的毒性效应

通过研究壬基酚对黑斑蛙(Rana nigromaculata)血浆渗透压以及血细胞的影响,探讨壬基酚对黑斑蛙血液的毒性效应。用200、400和600mg/kg壬基酚分别对黑斑蛙腹部淋巴囊注射染毒,在不同的时间间隔内利用渗透压仪测量各组血浆渗透压,同时制作血涂片观察血细胞的异常现象。结果表明,在相同处理时间内,随着壬基酚浓度的增加,黑斑蛙血浆渗透压值上升,血细胞膨大,血细胞核分裂以及核质不均匀现象明显;在相同浓度处理组中,随着处理时间的延长,黑斑蛙血浆渗透压上升,血细胞膨大,细胞核损害严重。壬基酚可诱发红细胞出现微核现象,随着壬基酚浓度的增加,同一处理时间内黑斑蛙红细胞微核及核异常率呈现先上升后下降的变化规律;随着处理时间的延长,各处理组红细胞微核率及核异常率呈现下降的趋势。     

小鼠腹腔注射甲醛后骨髓细胞微核变化检测

本实验目的是检测不同剂量甲醛经腹腔注射染毒小鼠后,小鼠骨髓嗜多染红细胞数量和微核的数量与注射剂量之间的关系。结果甲醛剂量和嗜多染红细胞数、微核数、微核率(‰)均呈正相关。     

城镇污水诱发青蛙蝌蚪红细胞微核及其在环境监测中的应用

本文利用青蛙(Rana nigromaculata Hallowell)蝌蚪红细胞微核试验,作为检测城镇污水诱变活性的一种新的监测技术。在16d生活污水处理的实验中,青蛙蝌蚪红细胞微核细胞率2d后就呈现统计上的显著增加,并随处理时间的延长而增高,第12d达到最大值。在不同浓度混合污水处理实验中,蝌蚪红细胞微核细胞率呈现明显的剂量依赖性增加。上述实验证明城镇生活污水和混合污水都具有较强的诱变活性。作者从遗传毒理学的角度评价了湖北黄州综合生物塘系统对污水诱变活性的净化效能。城镇混合污水经综合生物塘各级塘处理,蝌蚪微核细胞率逐级下降,由进水的7.54‰下降到最后出水的1.52‰,接近对照(1.07‰)水平。其中综合生物塘的藻菌单元比水生植物单元对污水诱变活性具有更强的净化效力。本文提出污水“诱变指数”可作为综合生物塘一项功能评价指标。     

除草剂精禾草克对黄鳝细胞遗传毒性的研究

采用红细胞微核和核异常、染色体数目和结构畸变的方法,研究了除草剂精禾草克对黄鳝细胞的遗传毒性。结果表明,不同浓度的精禾草克作用３０小时,红细胞微核细胞率没有明显变化,核异常细胞率和总核异常细胞率均有所上升,部分组与对照组差异显著。染色体数目畸变率均有所上升,有的组甚至与对照组差异级显著,染色体结构畸变率也显著上升,各组与对照差异显著或极显著。表明一定浓度范围内的精禾草克作用一定时间对黄鳝在明显的遗     

Effects of Roundup Herbicide and Increase in Water Temperature on the Parameters of Peripheral Blood Cells in Amur Sleeper <Emphasis Type="Italic">Perccottus glenii</Emphasis> Dybowski

The separate and combined effects of chronic 30-day exposure to the herbicide Roundup in a sublethal concentration of 2 μg/L and an increase in water temperature at a rate of 8°C/h on the parameters of red and white blood in juveniles of Amur sleeper Perccottus glenii Dybowski have been studied. The ratio of mature and immature erythrocytes in the peripheral blood do not change under the influence of the studied factors. An increase in temperature after chronic exposure to Roundup leads to a decrease in red blood cell sizes and increase in the share of abnormal cells. Exposure to the herbicide and the rise in water temperature have the opposite effect on the number of amitosis in erythrocytes and the ratio of leucocyte cells; an antagonistic effect is identified under the combined action of the factors. Changes in white blood correspond to a nonspecific stress response; changes in red blood indicate a reduction in compensatory responses to hypoxia.     

不同性别黄鳝六种组织中LDH同工酶电泳谱的初步研究

本文报告了运用聚丙烯酰胺凝胶圆盘电泳研究不同性别黄鳝的血清、心肌、骨骼肌、肝、肾和生殖腺等六种组织器官中ＬＤＨ同工酶。结果表明六种不同组织中ＬＤＨ同工酶谱各不相同,具有明显的组织特异性。在不同性别中某些同一种组织的ＬＤＨ同工酶谱也发生变化,这说明决定黄鳝ＬＤＨ同工酶表达的因素在不同性别中有差异。     

汞离子及铬离子对黄鳝脾免疫细胞的影响

采用毒性实验方法,用不同浓度的汞离子(Hg2+)、铬离子(Cr6+)分别处理黄鳝(Monopterusalbus),经1、2、4、8 d后,通过光镜观察黄鳝脾组织结构及免疫细胞数量的变化。结果表明,对照组黄鳝脾被膜较薄,未见明显的小梁,实质由红髓和白髓构成。白髓中淋巴细胞聚集成群,未见明显脾小结,但可见动脉周围淋巴鞘。红髓由脾索与脾窦组成。脾中有椭圆体,其末端向脾髓开放。黑色素巨噬细胞中心形成。经两种重金属离子分别染毒后的黄鳝脾与对照组相比,组织结构表现出相似的变化,即随着重金属离子浓度的增加和染毒时间的延长,脾组织中的黑色素巨噬细胞中心逐渐增大、增多,最后减少;黑色素巨噬细胞先增加后减少。淋巴组织逐渐松散,排列稀疏混乱,淋巴细胞界限逐渐不清晰,呈退化趋势,数量先增加后减少。粒细胞数量的变化趋势与淋巴细胞一致。红血细胞大量破坏,血窦扩张。     

Density-gradient enrichment of newly-formed mouse erythrocytes: Application to the micronucleus test

To facilitate scoring micronuclei in peripheral blood erythrocytes, we have developed a centrifugation method to concentrate polychromatic and newly-formed normochromatic erythrocytes from microliter quantities of blood in a Percoll density gradient. Erythrocytes were separated into two discrete bands in a continuous gradient generated in situ in a microhematocrit capillary tube. The upper band contained white blood cells and a mixture of polychromatic and young normochromatic erythrocytes with a density of 1.080–1.082 g/ml. More than 75% of the polychromatic erythrocytes in samples of normal blood were recovered in the upper band. Older normochromatic erythrocytes migrated to the lower band. The frequency of polychromatic erythrocytes was increased from approximately 2% in whole blood to 60–80% in the upper band. After clastogen treatments, the elevated frequencies of micronuclei in the upper band polychromatic erythrocytes were similar to those in unfractionated blood. The frequencies of micronucleated normochromatic erythrocytes in the upper band were higher than those in whole blood at 48, 72 and 96 h after clastogen treatment, consistent with the expectation that the low-density normochromatic cells are newly derived from polychromatic erythrocytes. This density-gradient centrifugation technique enhances the efficiency of scoring micronuclei in the acute peripheral blood micronucleus test.     

Density-gradient enrichment of newly-formed mouse erythrocytes. Application to the micronucleus test

To facilitate scoring micronuclei in peripheral blood erythrocytes, we have developed a centrifugation method to concentrate polychromatic and newly-formed normochromatic erythrocytes from microliter quantities of blood in a Percoll density gradient. Erythrocytes were separated into two discrete bands in a continuous gradient generated in situ in a microhematocrit capillary tube. The upper band contained white blood cells and a mixture of polychromatic and young normochromatic erythrocytes with a density of 1.080-1.082 g/ml. More than 75% of the polychromatic erythrocytes in samples of normal blood were recovered in the upper band. Older normochromatic erythrocytes migrated to the lower band. The frequency of polychromatic erythrocytes was increased from approximately 2% in whole blood to 60-80% in the upper band. After clastogen treatments, the elevated frequencies of micronuclei in the upper band polychromatic erythrocytes were similar to those in unfractionated blood. The frequencies of micronucleated normochromatic erythrocytes in the upper band were higher than those in whole blood at 48, 72 and 96 h after clastogen treatment, consistent with the expectation that the low-density normochromatic cells are newly derived from polychromatic erythrocytes. This density-gradient centrifugation technique enhances the efficiency of scoring micronuclei in the acute peripheral blood micronucleus test.     

Evaluation of nongenotoxic and genotoxic factors modulating the frequency of micronucleated erythrocytes in the peripheral blood of mice

The hematological micronucleus test is regarded as an indicator of the clastogenic effect of chemicals and acute cytogenetic damage. The test can be carried out in red blood cells of the bone marrow and of the spleen, as well as in peripheral erythrocytes. We have determined the precise background values of micronucleated red blood cells for the peripheral blood of BALB/c, DBA/2, and NMRI mice. Bleeding, phenylhydrazine-induced hemolysis, and splenectomy generated an increase of micronucleated erythrocytes in the peripheral blood of mice. Our data thus demonstrate that such factors should be taken into consideration when the micronucleus test is used for screening the genotoxic potential of chemicals. Furthermore, the micronucleus-inducing effect of cyclophosphamide was studied in normal and splenectomized mice and, in addition, a comparison of the sensitivity of the micronucleus test was carried out in peripheral blood and bone marrow after cyclophosphamide treatment. Our data demonstrate that the kinetics of micronucleus formation were similar in normal and in splenectomized mice in which the micronucleus levels had returned to normal. The comparison of micronucleus formation in bone marrow and peripheral blood after cyclophosphamide treatment revealed the generation of similar quantities of micronucleated red blood cells in both tissues. The physiological mechanisms of micronucleus formation and removal and the potential role of chemically induced spleen damage during this process are discussed; the usefulness of the peripheral micronucleus test as a simple, rapid, and animal-saving modification of the standard bone marrow test is evaluated.Abbreviations CP cyclophosphamide - MN micronuclei - MNCE micronucleated normochromatic erythrocytes - MNPCE micronucleated polychromatic erythrocytes - MNRBC micronucleated red blood cells - NCE normochromatic erythrocytes - PCE polychromatic erythrocytes     

Evaluation of nongenotoxic and genotoxic factors modulating the frequency of micronucleated erythrocytes in the peripheral blood of mice

The hematological micronucleus test is regarded as an indicator of the clastogenic effect of chemicals and acute cytogenetic damage. The test can be carried out in red blood cells of the bone marrow and of the spleen, as well as in peripheral erythrocytes. We have determined the precise background values of micronucleated red blood cells for the peripheral blood of BALB/c DBA/2, and NMRI mice. Bleeding, phenylhydrazine-induced hemolysis, and splenectomy generated an increase of micronucleated erythrocytes in the peripheral blood of mice. Our data thus demonstrate that such factors should be taken into consideration when the micronucleus test is used for screening the genotoxic potential of chemicals. Furthermore, the micronucleus-inducing effect of cyclophosphamide was studied in normal and splenectomized mice and, in addition, a comparison of the sensitivity of the micronucleus test was carried out in peripheral blood and bone marrow after cyclophosphamide treatment. Our data demonstrate that the kinetics of micronucleus formation were similar in normal and in splenectomized mice in which the micronucleus levels had returned to normal. The comparison of micronucleus formation in bone marrow and peripheral blood after cyclophosphamide treatment revealed the generation of similar quantities of micronucleated red blood cells in both tissues. The physiological mechanisms of micronucleus formation and removal and the potential role of chemically induced spleen damage during this process are discussed; the usefulness of the peripheral micronucleus test as a simple, rapid, and animal-saving modification of the standard bone marrow test is evaluated.Abbreviations CP cyclophosphamide - MN micronuclei - MNCE micronucleated normochromatic erythrocytes - MNPCE micronucleated polychromatic erythrocytes - MNRBC micronucleated red blood cells - NCE normochromatic erythrocytes - PCE polychromatic erythrocytes     

Simplified mouse peripheral reticulocyte micronucleus test with dimethylnitrosamine.

The induction of micronuclei by treatment with dimethylnitrosamine was evaluated and compared in peripheral blood and bone marrow cells of male CD-1 mice. Peripheral blood preparations were made on acridine orange (AO)-coated slides and scanned by fluorescence microscopy. A significant increase in micronuclei was observed 24 h after treatment in bone marrow polychromatic erythrocytes, and 24-48 h after treatment in peripheral reticulocytes. The peak frequency of micronuclei in peripheral reticulocytes was delayed by about 24 h relative to bone marrow polychromatic erythrocytes. This micronucleus test using peripheral blood was shown to be easy to do and as sensitive as the test using bone marrow cells. From this result, it is concluded that the method with AO-coated slides and peripheral blood is as suitable as bone marrow cells for the micronucleus assay.