镉染毒小鼠网织红细胞微核流式细胞术研究

目的通过流式细胞术(FCM)检测外周血微核方法，研究氯化镉(CdCl2)染毒小鼠的外周血含微核网织红细胞率，评价流式细胞术在微核检测中的应用价值及探讨氯化镉的致突变作用。方法采用转铁蛋白受体CD71荧光抗体和碘化丙啶(PI)染色，并以疟原虫感染小鼠外周血红细胞为微核模式生物调校流式细肛仪．检测10～60mg／kg环磷酰胺染毒和50～400μg／kg氯化镉染毒对小鼠的外周血含微核网织红细胞率的变化。结果不同剂量环磷酰胺染毒小鼠的外周血含微核网织红细胞率和骨髓含微核网织红细胞率均明显高于对照组小鼠，且二者之间相关性良好(r=0．955)；不同剂量氯化镉染毒小鼠外周血含微核网织红细胞率未见明显升高。结论FCM测定外周血含微核网织红细胞率可替代传统显微镜检查骨髓含微核网织红细胞率用于遗传毒性评价；在本研究条件下未观察到氯化镉有明显的诱导小鼠外周血含微核网织红细胞形成的作用。     

CD71荧光抗体和PI染色的微核流式细胞术检测研究

目的探索CD71荧光抗体和PI染色的微核流式细胞术自动化检测方法。方法采用-85℃甲醇固定细胞,CD71荧光抗体和PI染色,以伯氏疟原虫感染小鼠血细胞为微核模式生物调校流式细胞仪,建立微核流式细胞术检测方法,并对秋水仙素(COL)诱导的小鼠外周血含微核网织红细胞进行了检测。结果所建立的方法能明确分辨外周血含与不含微核的网织红细胞和成熟红细胞,COL处理小鼠的外周血含微核网织红细胞率呈明显的剂量依赖性升高,流式细胞术与显微镜检测的外周血含微核网织红细胞率有良好相关性(r=098)。结论微核流式细胞术检测方法能准确地检测小鼠外周血含微核网织红细胞。     

铁路危险品货运站空气颗粒污染物有机提取物的小鼠外周血网织红细胞微核试验

目的探讨铁路危险品货运站空气颗粒污染物对小鼠外周血网织红细胞微核的诱导作用.方法选用雄性BALB/c小鼠随机分为实验组和对照组,空气颗粒污染物的丙酮提取物按20、40、80、160 mg/kg的剂量,以腹腔注射方式一次性染毒.同时进行低剂量(10 mg/kg)腹腔注射重复式染毒,每天1次共5天,于染毒后24、48、72 h采集尾血进行网织红细胞微核分析.结果外周血网织红细胞微核率呈现出良好的时间-反应关系,微核的高峰出现在染毒后48 h,此时各剂量组微核率均高于对照组.低剂量重复染毒试验结果表明,在24～72 h内网织红细胞微核率始终处于较高水平,与对照组相比差异有显著性(P＜0.05).结论铁路危险品货运站空气颗粒污染物对雄性小鼠外周血网织红细胞微核具有明显的诱导作用.     

铁路危险品货运站空气颗粒污染有机提取物的小鼠外周血网织红细胞微核试验

目的 探讨铁路危险品货运站空气颗粒污染物对小鼠外周血网织红细胞微核的诱导作用。方法 选用雌性BALB／c小鼠随机分为实验组和对照组,空气颗粒污染物的丙酮提取物按20、40、80、160mg/kg的剂量,以腹腔注射方式一次性染毒。同时进行低剂量（10mg/kg）腹腔注射重复式染毒,每天1次共5天,于染毒后24、48、72h采集尾血进行网织红细胞微核分析。结果 外周血网织红细胞微核率呈现出良好的时间－反应关系,微核的高峰出现在染毒后47h,此时各剂量组微核率均高于对照组。低剂量重复染毒试验结果表明,在24－72h内网织红细胞微核率始终处于较高水平,与对照组相比差异有显著性（P＜0．05）。结论 铁路危险品货运站空气颗粒污染物对雌性小鼠外周血网织红细胞微核具有明显的诱导作用。     

DNA Damage among Peripheral Lymphocytes Induced by Phoxim Exposure in Mice

目的 探讨辛硫磷对小鼠的遗传毒性.方法 将60只健康SPF级昆明小鼠随机分为6组,分别为阴性对照(生理盐水)组和10、20、40、80mg/kg辛硫磷染毒组以及阳性对照组(40mg/kg环磷酰胺腹腔注射,每日1次,连续2d),每组10只,雌雄各半.采用灌胃方式进行染毒,染毒容量为10ml/kg,每日1次,连续染毒14d.采用微核试验和单细胞凝胶电泳试验检测辛硫磷的遗传毒性.结果 与阴性对照组比较,仅40和80mg/kg辛硫磷染毒组小鼠的骨髓细胞微核率显著性升高,各剂量辛硫磷染毒组小鼠外周血淋巴细胞的拖尾率和DNA损伤专用单位均升高,差异有统计学意义(P＜0.05).随着辛硫磷染毒剂量的升高,小鼠骨髓细胞微核率、外周血淋巴细胞的拖尾率和DNA损伤专用单位均呈上升趋势.结论 辛硫磷对小鼠具有遗传毒性.     

辛硫磷对小鼠外周血淋巴细胞DNA损伤的研究

目的探讨辛硫磷对小鼠的遗传毒性。方法将60只健康SPF级昆明小鼠随机分为6组,分别为阴性对照(生理盐水)组和10、20、40、80 mg/kg辛硫磷染毒组以及阳性对照组(40 mg/kg环磷酰胺腹腔注射,每日1次,连续2 d),每组10只,雌雄各半。采用灌胃方式进行染毒,染毒容量为10 ml/kg,每日1次,连续染毒14 d。采用微核试验和单细胞凝胶电泳试验检测辛硫磷的遗传毒性。结果与阴性对照组比较,仅40和80 mg/kg辛硫磷染毒组小鼠的骨髓细胞微核率显著性升高,各剂量辛硫磷染毒组小鼠外周血淋巴细胞的拖尾率和DNA损伤专用单位均升高,差异有统计学意义(P<0.05)。随着辛硫磷染毒剂量的升高,小鼠骨髓细胞微核率、外周血淋巴细胞的拖尾率和DNA损伤专用单位均呈上升趋势。结论辛硫磷对小鼠具有遗传毒性。     

有机磷农药对小鼠骨髓细胞遗传物质的影响

目的探讨较长时间接触有机磷农药对动物骨髓细胞遗传物质的影响,为有机磷农药对农副产品污染的综合防治提供科学依据。方法选择昆明种成年小鼠经口灌胃染毒,1次/d,连续染毒21 d。敌敌畏(DDVP)染毒剂量为5.0,10.0和20.0 mg/kg,氧乐果染毒剂量为2.5,5.0和10.0 mg/kg,对照组给予等容积的生理盐水。观察染毒21 d后小鼠骨髓嗜多染红细胞(PCE)微核率和染色体畸变细胞率的变化。结果各染毒组微核率和染色体畸变细胞率均显著高于对照组(P<0.05或P<0.01)。结论有机磷农药的长期毒性作用可引起小鼠骨髓细胞遗传物质的损伤,导致遗传毒性效应。     

虾青素对辛硫磷所致小鼠DNA损伤的拮抗作用

目的 探讨虾青素对辛硫磷所致小鼠遗传毒性的拮抗作用.方法 将50只健康SPF级昆明小鼠随机分为5组,分别为对照组和80 mg/kg辛硫磷染毒组以及低、中、高剂量虾青素保护组(2mg/kg、4mg/kg、8 mg/kg),每组10只,雌雄各半.早晨低、中、高剂量虾青素保护组分别灌胃染毒2、4、8 mg/kg虾青素,对照组和辛硫磷染毒组灌胃染毒橄榄油,染毒容量为10 ml/kg;间隔约8h后,下午低、中、高剂量虾青素保护组和辛硫磷染毒组分别灌胃染毒80mg/kg辛硫磷,对照组灌胃生理盐水,染毒容量为10 ml/kg,连续染毒14d.采用微核实验检测小鼠骨髓细胞微核率,采用单细胞凝胶电泳实验检测小鼠外周血淋巴细胞的DNA损伤.结果 虾青素干预能抑制辛硫磷染毒所致的小鼠骨髓细胞微核率及外周血淋巴细胞拖尾率和DNA损伤专用单位的升高;且随着虾青素剂量的升高,小鼠的骨髓细胞微核率、外周血淋巴细胞的拖尾率和DNA损伤专用单位均呈下降趋势.结论 虾青素对辛硫磷所致小鼠骨髓细胞微核率性的升高及外周血淋巴细胞的DNA损伤具有明显的拮抗作用.     

非整倍体毒剂和染色体断裂剂的FCM微核检测区分方法研究

目的研究流式细胞术（flow cytometry，FCM）微核检测筛选非整倍体毒剂和染色体断裂剂的方法。方法采用CD71荧光抗体和碘丙啶（PI）双染色，通过FCM检测环磷酰胺（CP）和秋水仙素（COL）诱导的NIH小鼠外周血含微核网织红细胞的PI荧光强度，比较它们与二倍体细胞PI荧光强度中位数的比值，以探索通过FCM微核检测筛选非整倍体毒剂和染色体断裂剂的方法。结果染色体断裂剂CP诱导的含微核网织红细胞与二倍体细胞PI荧光强度中位数的比值较低，非整倍体毒剂COL诱导的含微核网织红细胞与二倍体细胞PI荧光强度中位数的比值较高，两者间差异有统计学意义（P〈0．01）。结论通过FCM检测微核PI染色强度，可初步判断诱导微核形成的因素是否具有染色体断裂剂和非整倍体毒剂毒性。     

氯化镍对小鼠生殖毒性的影响

对探讨氯化镍（NiCl2)对雌性小鼠（F0及F1）生殖能力的影响,采用两代生殖毒性实验,依NiCl2注射剂量将雌性小鼠分为5mg/kg和10mg/kg两组,腹腔注射10天后使其交配。结果显示F0、F1染毒组小鼠的生殖能力明显低于对照组。说明NiCl2不仅影响F0代雌性小鼠的生殖能力,还对F1代雌性小鼠有性腺毒作用的远期效应。     

秋水仙素和环磷酰胺诱发微核的多色荧光原位杂交研究

目的 分析秋水仙素和环磷酰胺诱导的小鼠骨髓红细胞微核的染色体组成。方法 用着丝粒和端粒ＤＮＡ探针多色荧光原位杂交检测微核的染色体组成。结果 水仙素诱导的微核８３．５％既含着丝粒信号又含端粒信号,环磷酰胺诱导的微核７４．５％只含端粒信号。结论秋水仙素诱导的微核主要由整条染色体组成,环磷酰胺诱导的微核则主要由染色体继片组成。着丝粒和端粒ＤＮＡ探针多色荧光原位杂交是一种较为精确的分析微核染色体组成的方法     

Induction of micronuclei in somatic cells of mice exposed to x-rays or nonylphenol and to a combination of both agents

The effects of X-rays, nonylphenol (NL) and combination of both agents on the induction of micronuclei in mouse somatic cells were investigated. Pzh: SFIS mice were exposed during 2 weeks, 5 days per week to X-rays (doses: 0.05 Gy, 0.10 Gy,0.20 Gy), nonylphenol (doses: 25 mg/kg bw NL, 50 mg/kg bw NL, 100 mg/kg bw NL) and to a combination of X-rays and nonylphenol (doses: 0.05 Gy + 25 mg/kg bw NL, 0.10 Gy + 50 mg/kg bw NL). Samples from peripheral blood were taken 1 week after the start of exposure and 24 h after the end of exposure, whereas samples from bone marrow were taken 24 h after the end of exposure. Results obtained show that ionizing radiation, nonylphenol and combination of X-rays-NL in low doses induced micronuclei in peripheral blood and bone marrow reticulocytes. In contrast combined exposure to higher doses of both agents caused reduction frequency of micronuclei in the comparison to effects of X-rays acting alone. In bone marrow polychromatic erythrocytes the induction of micronuclei was enhanced after combined exposure to both agents in lower and higher doses.     

Comparison of 4', 6'-diamidino-2-phenylindole and Giemsa stainings in preimplantation mouse embryos micronucleus assay including a triple dose study

Analysis of micronuclei (MN) in preimplantation embryos is a good method for the evaluation of cytogenetic damage induced by occupational and environmental mutagen during early pregnancy. To examine whether conventional Giemsa staining produced the same accuracy of micronuclei as the DNA-specific 4', 6'-diamidino-2-phenylindole (DAPI) staining in preimplantation embryo induced by maternal exposure to chlorpyrifos, we conducted assays on 469 mouse (3 groups) preimplantation embryos micronucleus. Slides were stained with DAPI. After DAPI staining, the slides were de-stained and restained with Giemsa. Giemsa staining showed similar frequencies in MN to DNA-specific DAPI staining in all three groups. Both staining techniques revealed significant increases in frequency of MN in the treated group in comparison to the control group. Both methods showed a statistically significant correlation between MN frequency and the dose of chlorpyrifos. Compared with DAPI staining, the sensitivity of Giemsa staining was 85.0%, 86.0% and 90.9% for control, 40 mg/kg, and 80 mg/kg chlorpyrifos treated group, respectively. The specificity was 97.9%, 91.4% and 96.5% for control, 40 mg/kg, and 80 mg/kg chlorpyrifos treated group, respectively. Thus, we recommend that Giemsa staining technique be a standard staining method in detecting MN of preimplantation embryos induced by occupational or environmental hazards.     

猕猴桃汁抗环磷酰胺致突变作用的机理

目的用大鼠外周血双核淋巴细胞微核测试法（ＣＢＭＮＴ），在哺乳动物整体水平，研究猕猴桃汁抗环磷酰胺（ＣＰ）的致突变作用以及生物转化Ⅱ相酶的作用。方法测定大鼠外周血双核淋巴细胞微核细胞率及肝组织中总谷胱甘肽硫转移酶（ＧＳＴ）、尿苷二磷酸葡萄糖醛酸转移酶（ＵＤＰＧＴ）、谷氨酰胺转肽酶（γＧＴ）活性。结果猕猴桃汁对ＣＰ诱发的大鼠外周血双核淋巴细胞微核细胞率有显著抑制作用，能明显诱导大鼠肝脏总ＧＳＴ、ＵＤＧＴＰ活性，但对γＧＴ活性无显著影响。大鼠外周血双核淋巴细胞微核细胞率与总ＧＳＴ、ＵＤＧＴＰ活性呈明显负相关。结论猕猴桃汁抗ＣＰ致微核形成作用的机理可能是通过诱导机体外来化合物代谢解毒酶系，加速ＣＰ的代谢灭活     

Synthesis and in vivo anti-mutagenic activity of novel melatonin derivatives

Extensive literature suggests that melatonin play a role against the degenerative effect of central neurotoxins by its acting as free radical scavenger. This study aimed at evaluation of the anti-mutagenic activity of novel synthesized indole derivatives 2, 4a, and 8 in albino male mice in comparison with the parent melatonin. Efficacy of melatonin and its derivatives to influence cyclophosphamide (CP)-induced genotoxicity was tested using micronuclei (MN) formation in the bone marrow cells and determination of DNA, RNA and protein levels as well as cholinesterase and peroxidase activities in several organs of male mice. Following intragastrical injection of melatonin or one of its derivatives daily for 1 week, CP was given intraperitoneally, i.p., as a single dose of 25mg/kg BW. Pyridazin-4-yl thiadiazoloindole derivative 8, diaminothiophen-5-yl thiadiazoloindole derivative 4a and melatonin were significantly able to reduce the number of micronucleated polychromatic erythrocytes (MnPCEs) in the bone marrow cells induced by CP (P<0.0001, P<0.001, P<0.01, respectively). However, reduction of MN formation in the bone marrow cells was not significant when thiadiazoloindole derivative 2 was administered (P=0.14). Examination of the protective effect of melatonin and its derivatives on the levels of DNA, RNA and protein as well as enzyme activities showed that compound 8 had the ability to inhibit the clastogenic effect of CP in several organs of male mice. These findings suggest that compounds 4a, 8 and melatonin were able to reduce the mutagenicity effect of CP in male mice. The ability of compounds 4a, 8 and melatonin to reduce CP-related genotoxicity is possibly attributed to their antioxidant activity.     

母鼠有机磷农药暴露对着床前期小鼠胚胎的细胞遗传学及生长发育的影响

[目的]研究妊娠初期有机磷农药暴露对母体生殖功能，对子代细胞遗传学及生长发育的影响。[方法]以有机磷农药敌百虫为受试物，以着床期胚胎为研究对象，以反映染色体异常的微核形成和反映胚胎生长发育状况的构成细胞数为指标。孕鼠腹腔注射100或200mg/kg剂量的敌百虫溶液，3d后，在体视显微镜下从其子宫中取出胚胎，进行其形态学分析，微核试验及胚胎构成细胞数的分析。[结果]敌百虫暴露组的胚胎微核发生率明显高于对照组，分别为200mg／kg暴露组为16．9％，100mg／kg暴露组16．1％，对照组为5．3％，并有明显的剂量-反应关系。此外，母鼠暴露于敌百虫，可造成子代生长发育迟缓，表现为胚胎构成细胞数明显下降(200mg/kg组44．1，对照组54．7)。[结论]妊娠初期有机磷农药暴露对着床前期胚胎的细胞遗传学及生长发育有明显影响。着床前期小鼠胚胎的微核试验是评价母体早期暴露有机磷农药导致子代细胞遗传学改变的简便、灵敏的指标，本结果为进一步对着床后胚胎的研究，奠定了基础。     

Immunologic effects of nickel: I. Suppression of cellular and humoral immunity

The effects of nickel chloride on the cellular and humoral immune responses of mice were studied. A single intramuscular injection of nickel chloride (18.3 mg/kg) caused a significant involution of the thymus within 2 days following treatment. Significant reductions in the in vitro mitogen-stimulated response of lymphocytes from nickel chloride-treated mice (24 hr following a single injection of 18.3 or 36.6 mg/kg) were observed for the T-cell mitogens phytohemagglutinin (PHA) and concanavalin A (Con A), and the B- and T-cell mitogen pokeweed mitogen (PWM) but not the B-cell mitogen lipopolysaccharide (LPS). Theta-positive but not Ig-positive spleen cells were significantly reduced in nickel-treated mice compared with controls. Significant suppression of the primary antibody response to the T-cell dependent antigen sheep red blood cells was observed following a single injection of 18.3 mg/kg NiCl2. Natural killer (NK) cell activity was significantly suppressed following a single injection of 18.3 mg/kg NiCl2. The administration of NiCl2 (18.3 mg/kg) also decreased the amount of endotoxin required to kill 50% of treated mice, although this was not statistically significant. In all cases the immunosuppressive effects of NiCl2 were found to be transient with responses returning to normal within a few days. No alteration in the response of mice immunized with the T-cell independent antigen polyvinylpyrrolidone was observed following treatment with nickel. Furthermore, the phagocytic capacity of resident peritoneal macrophages from nickel-treated mice was not significantly different from saline-injected mice. The results indicate that NiCl2 predominantly affects T-cell mediated immune responses and natural killer cells.