46,XY,dup(10q) in direct CVS preparation and mosaic 48,XXXY,dup(10q) in CVS long-term culture and fetal tissue

Chorionic villus sampling (CVS) was performed on a 40-year-old woman at 9 1/2 menstrual weeks because of advanced maternal age. The direct preparation showed 46,XY,dup(10)(q11.2q23.2). CVS long-term culture and fetal tissue revealed a rare additional abnormality: 48,XXXY,dup(10)(q11.2q23.2). This abnormality represented the major cell line (>85 per cent in 691 cells) in an (XY)/XXY/XXXY/(XXXXY) mosaic (all cell lines presumably bearing the dup(10q); the presence of XY and XXXXY cell lines is uncertain). To our knowledge, this is the first report of trisomy 10q11-q23 and of prenatally detected 48,XXXY in chorionic villi. The mosaic could have resulted from early post-zygotic non-disjunctions in a 46,XY,dup(10q) or 47,XXY,dup(10q) zygote. The results from DNA studies of four polymorphisms, mapped to Xp and Xq, support this theory. The literature on prenatally detected cases with sex chromosome tetrasomy and pentasomy and those with additional autosomal abnormalities is reviewed. The reported case underlines the problem of false-negative findings when only direct CVS preparations are karyotyped.     

Effects of plant species coexistence on soil enzyme activities and soil microbial community structure under Cd and Pb combined pollution

The relationship between plant species coexistence and soil microbial communities under heavy metal pollution has attracted much attention in ecology. However, whether plant species coexistence could o set the impacts of heavy metal combined pollution on soil microbial community structure and soil enzymes activities is not well studied. The modified ecological dose model and PCR-RAPD method were used to assess the e ects of two plant species coexistence on soil microbial community and enzymes activities subjected to Cd and Pb combined stress. The results indicated that monoculture and mixed culture would increased microbe populations under Cd and Pb combined stress, and the order of sensitivity of microbial community responding to heavy metal stress was: actinomycetes > bacteria > fungi. The respirations were significantly higher in planted soil than that in unplanted soil. The plant species coexistence could enhance soil enzyme activities under Cd and Pb combined. Furthermore, planted soil would be helpful to enhance soil genetic polymorphisms, but Cd and Pb pollution would cause a decrease on soil genetic polymorphisms. Mixed culture would increase the ecological dose 50% (ED50) values, and the ED50 values for soil enzyme activities decreased with increasing culture time. The dehydrogenase was most sensitive to metal addition and easily loses activity under low dose of heavy metal. However, it was di cult to fully inhibit the phoshpatase activity, and urease responded similarly with phosphatase.     

镉致仔猪睾丸支持细胞抗氧化酶活性降低及DNA损伤研究

为了研究镉对仔猪睾丸支持细胞毒性影响,以仔猪睾丸支持细胞为实验模型,采用二步酶消化法分离支持细胞进行培养,并通过油红O和免疫细胞化学方法进行鉴定,探讨了0、10、20、40、80μmo.lL-1氯化镉对支持细胞的毒性作用.结果表明,二步酶消化法能分离出纯度较高的支持细胞,10μmol·L-1以上的氯化镉对支持细胞的生长具有抑制作用,并能使支持细胞的丙二醛含量升高,超氧化物岐化酶、谷胱甘肽过氧化物酶活性下降,造成支持细胞DNA损伤,并随着氯化镉浓度的增大存在明显的剂量-效应关系.     

一种高效提取焦化废水活性污泥总DNA的方法

对焦化废水活性污泥中微生物建立高质量的总DNA提取方法是开展分子生态学研究的重要前提.通过反复冻融-蛋白酶K-SDS、溶菌酶-反复冻融-SDS及溶菌酶-反复冻融-蛋白酶K-SDS这3种综合方法对焦化废水活性污泥总DNA进行提取,以OD260/OD280、OD260/OD230、产率、片段完整性、片段大小5个指标来评价样品总DNA的提取效果.结果表明,溶菌酶-反复冻融-蛋白酶K-SDS法所提取的总DNA的OD260/OD280值约为1.8,产率为1.90～16.30 μg·g-1,片断完整性好,主带清晰,大小约为23 kb,其PCR反应抑制物少,能够直接进行16S rRNA基因的PCR扩增.溶菌酶-反复冻融-蛋白酶K-SDS法能够为焦化废水活性污泥中微生物的分子生态学研究提供高质量的总DNA.     

不同浓度甲醛致大鼠肝细胞DNA氧化损伤作用

为了探讨外源性化合物暴露致内脏细胞DNA氧化损伤程度的定量检测方法,尝试以8\|羟基脱氧鸟苷（8-hydroxy-2′-deoxyguanosine, 8-OHdG）为分子生物标志物,以甲醛为模式外源性化合物,以丙二醛（malondialdehyde, MDA）为参考指标,应用大鼠肝细胞悬液进行体外染毒实验研究.同时,实验设置甲醛染毒的终浓度分别为0、5、15、45μmol· L-1,在大鼠肝细胞悬液染毒1h后,分别测量了肝细胞悬液中的8-羟基脱氧鸟苷（8-OHdG）和丙二醛（MDA）含量.研究结果显示,随着甲醛浓度升高,大鼠肝细胞中8-OHdG和MDA含量均呈升高趋势（F=59.55,p<0.01;F=75.82,p<0.01）,高浓度组（45μmol·L-1）8-OHdG和MDA含量与对照组的差异均具有显著性（p<0.01）,中浓度组（15μmol·L-1）的这种差别也具有显著性（p<0.05, p<0.01）,低浓度组（5μmol·L-1）的这种差异没有显著性（p>0.05）.因此,8-OHdG不但可以用于血液和尿液样品的检测,而且采用本项研究摸索出的前处理方法可以很好地定量测定内脏细胞DNA氧化损伤的程度.     

Improved preparation and identification of aristolochic acid-DNA adducts by solid-phase extraction with liquid chromatography-tandem mass spectrometry

Aristolochic acid (AA) is a known nephrotoxin and potential carcinogen, which can form covalent DNA adducts after metabolic activation in vivo and in vitro. A simple method for preparation and characterization of aristolochic acid-DNA adducts was developed. Four AA-adducts were synthesized by a direct reaction of AAI/AAII with 2’-deoxynucleosides. The reaction mixture was first cleaned-up and pre-concentrated using solid phase extraction (SPE), and further purified by a reversed-phase high performance liquid chromatography (HPLC). By the application of developed SPE procedure, matrices and byproducts in reaction mixture could be greatly reduced and adducts of high purity (more than 94% as indicated by HPLC) were obtained. The purified AA-DNA adducts were identified and characterized with liquid-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-Q-TOF-MS/MS) and LC-Diode array detector-fluorescence (LC-DAD-FL) analysis. This work provides a robust tool for possible large-scale preparation of AA-DNA adduct standards, which can promote the further studies on carcinogenic and mutagenic mechanism of aristolochic acids.     

pH值对SBR生物反应器出水溶解态有机质含量与构成的影响

为探讨污水二级生化出水中有机物(EfOM)的构成特点和变化规律,采用配置生活污水,对不同pH值运行条件下, SBR生物反应器出水中的溶解态有机质(DOM)含量和构成进行了比较研究.结果表明,运行期间pH值的调节对SBR反应器出水DOM含量与构成的影响均较大.初始pH为6.5的SBR反应器, pH值降低为6.0后,出水DOC由原来的4.0mg·L-1不断升高,进一步提高pH值,能够起到一定的缓解作用;初始pH为8.0的SBR反应器,出水DOC从4.0mg·L-1快速降低并保持在2mg·L-1左右, pH值降低为6.5后, DOC变化不大.单宁酸和蛋白质、碳水化合物、DNA等微生物代谢产物(SMPs)是出水DOM的主要组成部分, pH值对SMPs生成的影响是一个长期的过程,运行过程中降低pH值,造成出水SMPs含量不断升高. pH值对整个循环过程中溶解态有机物降解情况的影响,在厌氧阶段发挥了主要作用,较高pH值有利于厌氧阶段有机物的降解,而好氧阶段的有机物降解受pH值影响不大.     

蒙脱土、高岭土和针铁矿对DNA吸附与解吸特征

采用平衡法研究了蒙脱土、高岭土和针铁矿在不同pH值和不同电解质溶液下的DNA吸附解吸特征.结果表明,在4种电解质体系中,尤其在溶液pH值>5.0时,3种矿物在吸附过DNA后,pH有不同程度上升,但CaCl2电解质体系中含针铁矿溶液的pH开始下降.针铁矿对DNA的吸附率大约为75%～91%,高岭土对DNA的吸附率从pH为3.5时的90%开始下降,至pH 6.0～7.0时的下降至大约18%～50%.在一价电解质体系,当溶液pH值为2.2～5.0时蒙脱土对DNA的吸附率大约为90%开始下降至25%～40%,但在二价电解质体系蒙脱土对DNA的吸附率大约为92%.Freundlich等温吸附方程能更好的拟合针铁矿、蒙脱土和高岭土对DNA的吸附.蒙脱土对DNA相对吸附容量在pH 3.5时大于针铁矿,而pH 5.0时小于针铁矿.以NaOAc为解吸剂,针铁矿、蒙脱土和高岭土上吸附DNA在不同pH时解吸率分别为0.1%～0.4%、63.7%～90.2%和29.7%～87.5%.以NaH2PO4为解吸剂时,这些矿物吸附DNA的解吸率分别为69.6%～78.7%、0.8%～7.0%和0.4%～2.2%.这表明针铁矿吸附DNA时键合作用较大,蒙脱土和高岭土静电引力较大,这是DNA在不同电荷类型矿物表面的吸附差异.     

高效氯氰菊酯对小鼠肝细胞的氧化损伤

为了探讨高效氯氰菊酯对生物体的氧化损伤,以昆明小鼠为受试体,高效氯氰菊酯按10、和40mg·kg20-13个剂量水平,灌胃染毒小鼠7d,并以肝匀浆测定活性氧自由基(ROS)、还原型谷胱甘肽(GSH)、丙二醛(MDA)含量,以肝细胞测定DNA-蛋白质交联(DPC)系数.实验结果表明,随着高效氯氰菊酯染毒剂量的升高,ROS和MDA含量及DPC系数逐渐上升,GSH含量逐渐降低,各指标呈一定的剂量-效应关系.染毒剂量≥20mg·kg-1时,处理组的ROS含量和DPC系数与对照组有显著差异(p<0.05);染毒剂量≥40mg·kg-1时,GSH和MDA含量与对照组有显著差异(p<0.05),DPC系数有极显著差异(p<0.01).说明较高剂量的高效氯氰菊酯可造成小鼠肝脏的氧化损伤和DNA-蛋白质交联作用增强.