Allele-specific amplification for preimplantation genetic diagnosis (PGD) of spinal muscular atrophy

We have developed a new allele-specific amplification method for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA; Werdnig-Hoffmann disease) from a single cell. This method is based on the detection of the deletion of exon 7 of the telomeric copy of the survival motor neurone (SMN^t) gene. An oligonucleotide was designed to be specific to the SMN^t nucleotidic sequence with exonic mismatch G (for SMN^t)→A (for SMN^c) at its 3′ end. This test produces reliable PCR products in 95% of single lymphoblasts (85/88) tested as well as in 16/16 blastomeres from normal controls. Specificity analysis showed that we were able to detect homozygous deletion of the SMN^t gene in 99% of single lymphoblasts (103/104) from a SMA patient. No contamination was detected in 68 blanks tested. Multiple cell and DNA dilution analysis revealed that the test is accurate and specific up to 100 pg DNA and should thus also be suitable for PGD at the blastocyst stage. This rapid procedure requires a single round of fluorescent PCR and no restriction digestion, while previously described single cell methods include nested PCR followed by restriction enzyme digestion. Two PGD cycles for SMA using this procedure were performed in our centre. Copyright © 2001 John Wiley & Sons, Ltd.     

DEHP经sirt1/pgc-1a通路诱导的HepG2细胞线粒体损伤效应

为研究邻苯二甲酸二（2-乙基己基）酯（DEHP）是否通过sirt1/pgc-1a通路对细胞产生损伤效应， 通过体外培养HepG2细胞，在1.6， 8， 40， 200， 1000mmol/L DEHP处理24或48h后，采用CCK-8测定细胞活力，ATP试剂盒检测细胞内ATP含量，NO试剂盒检测细胞上清NO含量，ELISA试剂盒检测细胞上清炎症因子TNF-a、IL-6的含量；同时在DEHP作用于HepG2细胞24或48h后，用Western blot检测线粒体调控基因sirt1、pgc-1a，nrf1、tfam的蛋白表达.结果表明：不同浓度的DEHP处理24， 48h后，细胞活力在高剂量都呈显著下降趋势.DEHP能引起ATP含量的显著下降、炎症因子含量的显著提高，但NO含量无显著变化.4种线粒体调控基因的蛋白表达水平sirt1、pgc-1a，nrf1、tfam在24h时无显著变化.而在48h时，随着剂量增加蛋白含量呈上升后下降趋势，并且sirt1在8mmol/L呈显著上升趋势，随后每个蛋白都在1000mmol/L显著下降（P < 0.05）.DEHP能引起HepG2细胞氧化应激，并通过影响sirt1/pgc-1a信号通路表达来影响线粒体生物合成从而造成细胞损伤.     

Toxicity screening of biodegradable polymers. II. Evaluation of cell culture test with medium extract

Cell culture testing with material extracts was applied to toxicity screening of some commercial degradable plastics: a plasticized cellulose acetate, an aliphatic polyester (Bionolle), polyhydroxybutyrate-co-hydroxyvalerate (Biopol), and polycaprolactone (TONE polymer). Cell culture medium with serum was used as extraction medium. Methods for the determination of morphology and viability of cells cultured in the extract were investigated. Phase-contrast light microscopy of cells, enhanced by neutral red staining, provides high-contrast images for qualitative evaluation of cell morphology and lysis. Compared to the determination of protein using the Bradford method and of neutral red uptake, the determination of dehydrogenase activity using 3-[4,5-dimethylthia-zol-2-yl]-2,5-diephenyl-tetrazolium bromide (MTT) is more sensitive and accurate. The relative MTT activity of cells cultured in fresh extracts indicate that TONE polymer (all shapes) and Bionolle (test bars and films) are comparable to materials currently used in the food industry (polyethylene terephthalate, atactic and isotactic polystyrene) with no toxic effects on cells.     

牛粪混合液微生物燃料电池长期运行稳定性研究

长期运行稳定性是微生物燃料电池(microbial fuel cells,MFCs)技术的一项重要特征,是其能否走向实际应用的关键.生物阴极MFC利用牛粪产电的长期运行特征的研究表明,该MFC可长期、高效、稳定地产电.在100Ω外电阻下,171 d的运行期内,电池功率密度平均为6.77 W·m-3±2.11 W·m-3.第70 d的电池开路电压、内阻、最高功率密度分别为0.874 V、22.1Ω和14.1 W·m-3.随着运行时间延长,每30 d的总化学需氧量(total chemical oxygen demand,TCOD)去除量不断递减,而库仑效率(Coulomb efficiency,CE)不断递增,在121~150 d,CE可达17.5%±3.3%.阳极微生物群落结构分析表明,Proteobacteria(45%)、Bacteroidetes(22%)、Firmicutes(17%)、Actinobacteria(11%)在阳极生物膜中占有优势地位.Clostridium、Cellulomonas等已被证明具有产电能力或纤维素降解能力的细菌是阳极生物膜中的关键功能种群.     

两种接种源微生物燃料电池同步降解刚果红与产电性能研究

分别以印染废水污泥和生活污水污泥接种微生物燃料电池(MFC-I和MFC-M),以刚果红为模型染料,研究了接种源对MFC同步脱色偶氮染料和产电性能的影响.结果表明,接种源对MFC同步降解刚果红与产电性能影响显著.产电方面,MFC-I的启动电压高于MFC-M,但稳定运行后二者最大输出电压差别不大.稳定运行阶段,MFC-M最大功率密度达到29.09 mW·m-2,比MFC-I高2.11倍;阳极极化阻抗(R ct)为748.9Ω,比MFC-I低38.0%.脱色方面,MFC-I对刚果红的脱色速率比MFC-M高14.4%~20.3%;但脱色产物一样,均为2-氨基—1,4-萘醌、2,2'-二氨基联苯、4,4'-二氨基联苯.经分离、纯化和鉴定,二者阳极均存在优势菌Pseudomonas sp.,但MFC-M阳极还存在优势菌Bacillus sp.,而MFC-I的阳极还存在优势菌Aquamicrobium sp..     

混菌接种条件下微生物燃料电池芯片的产电特性研究

微生物燃料电池(MFC)芯片因具有体积小、运行条件温和、产电稳定等优点而有可能成为一种新型的野外水环境监测系统中传感器供能方式.但目前采用纯菌种及贵重金属阳极构建的MFC芯片,不仅成本较高且纯菌种在复杂环境条件下不易存活和保持稳定.因此,本文通过采用混合菌群接种,以活性炭为阳极,构建了阳极体积为50μL的MFC芯片,发现其稳定运行最大输出电流为3.5μA,平均运行周期为8.0 h,最大输出功率约为160 nW,最大功率密度为10.2 mW·m-2.EIS分析结果表明,MFC芯片的总内阻约为35.6 kΩ,其中,阴阳极内阻占主要部分.本研究制备的MFC芯片产电性能达到了同类采用纯菌株及Au作阳极的MFC芯片的性能,表明采用低成本材料为阳极,接种混合菌液的MFC芯片是完全可行的.     

阳极内添加阳离子交换树脂提升辊压“三合一”膜电极MFC性能

阳极-隔膜-阴极的"三合一"膜电极结构能够最大程度减小阴阳极间距,提高微生物燃料电池(microbial fuel cell,MFC)的输出功率.为进一步提高MFC性能,本研究使用非贵金属材料构建了辊压"三合一"膜电极系统,其欧姆内阻降低至3~5Ω.以乙酸钠为底物,MFC的最高功率密度达到446 m W·m-2.向阳极内添加固体小球(如聚苯乙烯球和玻璃微球)可在辊压过程中增加阳极表面和内部的大孔,强化电解液向阴极的传递从而使MFC的功率密度提升10%.添加阳离子交换树脂能够进一步强化阳极内部的质子传递,提高阴极电位,从而将功率密度提升至543 m W·m-2.此外,阳极内添加阳离子交换树脂还可提高电池运行的稳定性和库仑效率.     

利用双室微生物燃料电池处理模拟废水的产电特性研究

本实验通过研究电池的启动过程、阳极有机物降解率和阴极Cu2+的去除率,评价了微生物燃料电池(microbial fuel cell,MFC)的产电和处理废水性能.以模拟糖蜜废水作为阳极基质,模拟电镀废水作阴极电子受体,建立简单的双室微生物燃料电池.结果表明在外电阻为800Ω的情况下,电池得到最大电压417.00 mV,从极化曲线上获得最大输出功率密度44.17mW.m-2,内阻为293Ω.电池在第五周期时,COD去除率也达到最高47.31%.在第四周期内,Cu2+最大的去除率为59.76%.综上所述,MFC在处理有机废水和电镀废水方面具有可行性.     

Investigation of nuclear enzyme topoisomerase as a putative molecular target of monohaloacetonitrile disinfection by-products

Disinfection by-products occur widely as the unintended effect of water disinfection and are associated with toxicity and adverse human health effects. Yet the molecular mechanisms of their toxicity are not well understood. To investigate the molecular basis of hyperploidy induction by monohaloacetonitriles, the interaction of monohaloacetonitriles with topoisomerase Ⅱ in Chinese hamster ovary cells was examined. We showed a concentration-dependent inhibition of DNA decatenation activity of topoisomerase under acellular conditions while in vitro monohaloacetonitrile treatment expressed mixed results. The working hypothesis, that topoisomerase Ⅱ is a molecular target of monohaloacetonitriles, was only partially supported.Nevertheless, this research serves as a starting point toward molecular mechanisms of toxic action of monohaloacetonitriles.     

以晶体硅太阳能电池产业为例的产业生命周期评价初探

面对日益复杂的环境问题和精细化环境管理需求,为了将生命周期评价在产业结构调整、发展方式转变中更好地发挥作用,对在产业层面开展生命周期评价的方法进行了探索研究.产业生命周期评价是在产品生命周期评价的基础上增加了:①基于“可拆解可组合”生态设计理念的功能单位和系统边界确定;②质量评估和数据整合的数据收集过程;③以不确定性分析来验证数据的合理性.选择晶体硅太阳能电池产业进行了产业生命周期评价的案例应用.结果表明:晶体硅太阳能电池产业可分为4个产品单元和11个工艺单元.基于上述产品单元和工艺单元的资源能源投入和污染物排放数据进行收集,在数据质量评估之后通过数据整合形成了产业生命周期数据清单.产业生命周期环境影响主要集中在呼吸系统影响（41.94%）、化石燃料（25.20%）、致癌（14.89%）和气候变化（8.80%）4个环境影响类别;减少环境影响的精准化途径是减少高纯多晶硅、硅片、电池片产品的电耗,组件产品中焊带消耗,硅片产品中的砂浆消耗和组件产品的铝合金边框消耗.蒙特卡洛分析结果显示,高纯多晶硅生命周期评价结果不确定性较高,与数据质量评估的结果较为一致.案例应用结果说明,产品生命周期评价可将生命周期评价从产品层面提升到产业层面,可为国家产业发展提供科学支撑.