The cytotoxicity of nici2 for mammalian cells in culture†

The effects of NiCl₂ were studied in two human cell lines, HeLa and diploid embryonic fibroblasts as well as in V79 Chinese hamster cells and in L‐A mouse fibroblasts. NiCl₂ produces a dose‐dependent depression of proliferation, mitotic rate, and viability, accompanied by an increasing release of lactic dehydrogenase and stimulation of lactic acid production. The plating efficiency is reduced, as are DNA and protein synthesis and, to a lesser degree, RNA synthesis.

The cytotoxicity of NiCl₂ is comparable in degree to those of PbCl₂ and MnCl₂, but is weaker than those of HgCl₂ and CdCl₂. However, the different sensitivities of different cell lines must also be considered.

NiCl₂ effects are more severe in serum‐free medium than in medium containing serum or serum albumin indicating that serum constituents, notably albumin, bind the metal effectively and inhibit cellular uptake; this confirms earlier reports on the serum binding and slow uptake of NiCl₂.

Synchronized cells are most sensitive in the Gl and early S phases of the cell cycle. In the Painter test the depression of DNA synthesis persists following cessation of exposure to NiCI₂. These findings contribute an explanation for the known genotoxic effects of nickel.     

Biochemical properties of beryllium potentially relevant to its carcinogenicity†

The carcinogenicity of beryllium to several animal species is well established and evidence exists which strongly suggests that this is the case in human exposure. In this review several biochemical properties of the metallocarcinogen are considered including, the causation of cell transformation, and infidelity of DNA synthesis, inhibition of cell division and enzyme induction, and interference with regulatory mechanisms controlling gene expression. These effects are discussed in relation to beryllium chemistry, cellular accumulation mechanisms and distribution to subcellular organdies and molecular targets. It is suggested that the ultimate location and interactions of the metal ion in cell nuclei and its selective inhibition of certain protein phosphorylation reactions in particular are the biochemical effects potentially most relevant to induction of beryllium carcinogenesis.     

Uptake and genotoxicity of micromolar concentrations of cobalt chloride in mammalian cells

Literature data concerning the genotoxicity of cobalt salts have been conflicting. To establish appropriate incubation conditions, we conducted a series of uptake studies, before genotoxicity was determined by DNA strand break induction in HeLa cells and mutagenicity in V79 Chinese hamster cells. Co(II) is taken up by HeLa cells in a concentration‐dependent manner and is accumulated inside the cell. The uptake is preceded by a fast association step to the outer membrane, with no saturation up to 24 h. DNA strand breaks as determined by nucleoid sedimentation are induced at concentrations as low as 50μMCoCl₂. The induction is time‐dependent, showing the highest number of breaks after 4h incubation with no further increase up to 24h. CoCl₂ is mutagenic at the HPRT‐locus, enhancing the spontaneous mutation frequency 4.2‐fold at 100μ?. Besides direct interactions with DNA, the mutagenicity of CoCl₂ could also be due to a decrease in the Fidelity of DNA polymerisation.     

壬基酚、五氯酚及硝基苯对端足类河蜾蠃蜚(Corophium acherusicum)的毒性效应

为检测海洋中环境激素及芳烃类化合物对端足类生物的污染危害,实验选择端足类河蜾蠃蜚(Corophium acherusicum)为受试生物,研究了其在壬基酚、五氯酚、硝基苯三种有机污染物暴露下的96 h急性致死毒性效应和7 d慢性DNA损伤毒性效应。计算获得壬基酚、五氯酚和硝基苯对河蜾蠃蜚的96 h半致死浓度(LC50)分别为70、465、25 000μg·L-1,三种有机污染物对河蜾蠃蜚的毒性强弱顺序为壬基酚五氯酚硝基苯。运用碱解旋法检测壬基酚、五氯酚和硝基苯对河蜾蠃蜚DNA损伤的程度,计算得到7 d半效应浓度(EC50)分别为30、256、11 000μg·L-1。实验结果表明:三种有机污染物浓度的不断加大,引起河蜾蠃蜚DNA损伤程度的不断增加,呈显著的剂量-效应关系。     

威海荣成鹿角菜野生种群遗传多样性及资源保护策略

利用随机扩增多态性DNA（RAPD）方法对威海荣成地区鹿角菜（Silvetiasiliquosa）野生种群共30个个体的遗传多样性水平及遗传结构进行研究。结果表明,利用21条随机引物共检测到112个多态性位点,多态性位点百分率为93．75％,Nei基因多样性指数为0．3515±0．1352,Shannon多样性指数为0．5046±0．1126。通过聚类分析而划为一类的3个亚种群间的遗传分化指数为0．1349～0．2189,基因流为1．7793。研究结果表明威海荣成地区的鹿角菜种群遗传多样性较高,种群内存在较为明显的遗传分化,种群遗传资源不受遗传漂变的影响。针对当地鹿角菜遗传资源的现状,应加强现存种群的就地保护,恢复种群规模;进行取样养殖保护时则需避免因近交而造成种群资源退化。     

不同浓度Cd~(2+)对鲤鱼基因组DNA的影响

鲤鱼(建鲤品种)幼鱼暴露于4个浓度组(0.005、0.05、0.5、5mg/L)的镉离子(Cd~(2+))2d,期间各组的个体均无死亡.用10个RAPD引物对处理后鲤鱼基因组进行扩增,亦未发现有明显的差异扩增条带.但在用甲基化敏感扩增多态性方法研究暴露后鲤鱼肌肉组织的基因组DNA甲基化的变化时发现,5mg/L的Cd~(2+)在d2会使鲤鱼基因组DNA中CCGG区域甲基化发生明显的变化.该组个体1mol后出现大量死亡.另外,0.05mg/L和0.5mg/L Cd~(2+)组个体基因组DNA中CCGG区域甲基化比例2d内略有下降,甲基化位点没有太大改变.说明鲤鱼幼鱼基因组CCGG区域的胞嘧啶甲基化比例在不同浓度Cd~(2+)处理中都比较稳定.低浓度Cd~(2+)条件下鲤鱼主要通过甲基化区域的微调减少Cd~(2+)的生理毒性.高浓度Cd~(2+)处理下,利于基因组DNA甲基化区域发生相当的变化,可能导致一些沉默基因的异常表达.图2表2参31     

氨氮对中华绒螯蟹仔蟹肝组织RNA/DNA及肝细胞结构的影响

以中华绒螯蟹（Eriocheir sinesis）仔蟹为研究对象。采用静水试验法,温度在20℃左右,用氨氮与过滤海水分设5个质量浓度组（2.04、6.35、9.04、11.75和19.13 mg.L-1）,同时以过滤海水为对照组（氨氮质量浓度为0.30 mg.L-1）,每一个质量浓度组设3个平行样本,对中华绒螯蟹仔蟹进行6 d胁迫试验。每2 d观察仔蟹的蜕壳情况,并分别于实验后的2、4和6 d取仔蟹肝组织样,应用激光共聚焦技术分析测定RNA/DNA荧光相素比,同时用透射电镜观察6 d后,最高浓度组和对照组仔蟹肝细胞结构的变化情况。试验结果表明;对于氨氮的胁迫,仔蟹的变态率随染毒浓度的增高而降低,同时变态蜕壳的时间也会产生延迟现象。氨氮的胁迫会引起仔蟹的肝组织细胞内的DNA和RNA的质量分数逐渐降低,导致RNA/DNA比率的不断下降。在高质量浓度氨氮的胁迫下,使得仔蟹肝细胞线粒体部分解体、胞质空泡化和染色质浓缩、转运泡数量增多,体积增大,细胞出现许多空泡和微绒毛消失等一系列的损伤变化。破坏了仔蟹自身机能正常的代谢和生长水平。     

Development of pretreatment protocol for DNA extraction from biofilm attached to biologic activated carbon (BAC) granules

The biologic activated carbon (BAC) process is widely used in drinking water treatments. A comprehensive molecular analysis of the microbial community structure provides very helpful data to improve the reactor performance. However, the bottleneck of deoxyribonucleic acid (DNA) extraction from BAC attached biofilm has to be solved since the conventional procedure was unsuccessful due to firm biomass attachment and adsorption capacity of the BAC granules. In this study, five pretreatments were compared, and adding skim milk followed by ultrasonic vibration was proven to be the optimal choice. This protocol was further tested using the vertical BAC samples from the full-scale biofilter of Pinghu Water Plant. The results showed the DNA yielded a range of 40 μg·g^-1 BAC (dry weight) to over 100 μg·g^-1 BAC (dry weight), which were consistent with the biomass distribution. All results suggested that the final protocol could produce qualified genomic DNA as a template from the BAC filter for downstream molecular biology researches.     

分子标记技术在石斛属植物中的应用研究进展

石斛作为一种名贵中药,近年来受到国内外研究人员的重视.本文对RFLP技术、位点特异性PCR、RAPD技术、AP-PCR技术、AFLP技术、ISSR技术、SRAP技术、DNA序列分析、基因芯片技术等DNA分子标记技术的基本原理和特点作了简要介绍,DNA分子标记技术为石斛类药材的鉴别提供了更加有效、可靠的鉴定方法.综述了近年来上述分子标记技术在石斛属植物中的应用研究进展,总结了这些技术在中药材鉴定中的地位,并对其发展前景做出了展望.     

北京市大气细颗粒物的遗传和非遗传毒性研究

用胞质阻断微核试验与单细胞凝胶电泳法检测北京市大气PM_2.5无机提取物与有机提取物对Balb/c 3T3细胞微核形成与DNA链断裂的影响;通过划痕染料示踪技术(SL/DT)观察PM_2.5无机提取物与有机提取物对Balb/c3T3细胞间通讯的影响.发现PM_2.5有机提取物可引起双核微核细胞率显著增加(P<0.01)及导致慧星细胞率和DNA迁移长度显著增加(P<0.01);PM_2.5有机提取物引起细胞间通讯的抑制; PM_2.5无机提取物未见明显毒作用.结果表明,PM_2.5可引起染色体损伤和原发性DNA损伤,抑制细胞间通讯,其毒作用主要由其有机成分引起.