Preimplantation genetic diagnosis of the fragile X syndrome by use of linked polymorphic markers

Fragile X syndrome is the most common cause of familial mental retardation. The most common mutation is expansion of a triplet (CGG)_n repeat in the 5′ untranslated region of the FMR1 gene on Xq27.3. The expansion is refractory to PCR due to preferential amplification of the smaller allele in heterozygous cells and the high GC content of the repeat and surrounding sequences. Direct detection of the normal parental alleles in preimplantation embryos has been used for preimplantation genetic diagnosis (PGD) of this disorder. However, this approach is only suitable for approximately 63% of couples due to the heterozygosity of the repeat in the normal population. As an alternative we investigated the use of polymorphic markers flanking the mutation to track the normal and premutation carrying maternal chromosomes in preimplantation embryos. Using a panel of 11 polymorphisms, six (CA)_n repeats and five single nucleotide polymorphisms, diagnosis was developed for 90% of referred couples. Multiplex amplification of informative markers was tested in 300 single buccal cells from interested couples with efficiency and allele drop out (ADO) rates ranging from 69% to 96% and 6% to 18%, respectively. Use of this approach is accurate and applicable to a larger number of patients at risk of transmitting fragile X to their offspring. Copyright © 2001 John Wiley & Sons, Ltd.     

A comparison of different lysis buffers to assess allele dropout from single cells for preimplantation genetic diagnosis

Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999 ). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (ΔF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. Copyright © 2001 John Wiley & Sons, Ltd.     

内质网应激在氟暴露致小鼠睾丸损伤中的作用

探讨了内质网应激在亚慢性氟暴露致小鼠睾丸损伤中的作用及分子机制.选用健康初断乳ICR雄性小鼠30只,随机分为对照组（C）、低氟组（LF）和高氟组（HF）,分别饮用自来水、5、30 mg·L^-1氟化钠水溶液90 d.亚慢性氟暴露结束后,以睾丸脏器系数、睾丸组织氧化/抗氧化酶和形态结构、精子质量、睾丸细胞凋亡、葡萄糖调节蛋白78（GRP78）、CCAAT/增强子结合蛋白同源蛋白（CHOP）、半胱氨酸天冬氨酸蛋白酶12（CASPASE-12）、半胱氨酸天冬氨酸蛋白酶3（CASPASE-3）为观测点.结果表明,与对照组比,LF组和HF组LDH、SOD、T-AOC活性下降,MDA含量上升,HF组GSH-PX活性下降,差异有统计学意义（p<0.05或p<0.01）;LF组可见细胞层次减少、间隙变大,成熟精子数量减少,HF组细胞溶解、层次紊乱,空泡化严重,少见成熟精子;LF组和HF组小鼠的精子活力降低,HF组小鼠精子数量下降,畸形率上升,差异有统计学意义（p<0.05或p<0.01）;LF组和HF组睾丸细胞凋亡指数上升,差异有统计学意义（p<0.01）;LF组和HF组Grp78、Caspase-12、Caspase-3基因表达水平上升,差异有统计学意义（p<0.05或p<0.01）.结果提示,除氧化应激以外,Caspase-12和Caspase-3基因表达异常可能是氟暴露致小鼠睾丸细胞凋亡异常的分子机制之一.     

石墨烯/聚苯胺修饰阴极对反硝化MFC性能影响

反硝化生物阴极微生物燃料电池（MFC）以电极为电子供体,在自养条件下完成硝酸盐去除过程.本研究以碳布（CC）为基底材料,分别制备获得还原氧化石墨烯修饰（rGO-CC）,聚苯胺修饰（PANI-CC）及二者复合修饰的CC电极（rGO/PANI-CC）,并考察其作为阴极材料对反硝化生物阴极MFC产电脱氮性能的影响.扫描电镜结果显示,rGO-CC和PANI-CC的碳纤维分别被片层状rGO和网状PANI覆盖,而rGO/PANI-CC表面呈现PANI在附着rGO的碳纤维上团聚的形貌,均增大了碳布的比表面积.循环伏安测试显示,rGO/PANI-CC具有最高的电化学活性.以rGO-CC,PANI-CC和rGO/PANI-CC为阴极构建MFC的产电能力分别提高了82%,24%和41%,其阴极对NO₃^--N的去除能力增强了23%,9%和13%.16S rDNA测序结果揭示修饰后电极表面微生物的多样性下降,Stappia和Pacacoccus属微生物的丰度增加.     

Performance of different macrophytes in the decontamination of and electricity generation from swine wastewater via an integrated constructed wetland-microbial fuel cell process

Plants constitute a major element of constructed wetlands(CWs).In this study,a coupled system comprising an integrated vertical flow CW(IVCW) and a microbial fuel cell(MFC) for swine wastewater tre atment was developed to research the effects of macrophytes commonly employed in CWs,Canna indica,Acorus calamus,and Ipomoea aquatica,on decontamination and electricity production in the system.Because of the different root types and amounts of oxygen released by the roots,the rates of chemical oxygen demand(COD) and ammonium nitrogen(NH_4~+-N) removal from the swine wastewater differed as well.In the unplanted,Canna indica,Acorus calamus,and Ipomoea aquatica systems,the COD removal rates were 80.20%,88.07%,84.70%,and 82.20%,respectively,and the NH_4~+-N removal rates were 49.96%,75.02%,70.25%,and 68.47%,respectively.The decontamination capability of the Canna indica system was better than those of the other systems.The average output voltages were 520±42,715±20,660±27,and 752±26 mV for the unplanted,Canna indica,Acorus calamus,and Ipomoea aquatica systems,respectively,and the maximum power densities were 0.2230,0.4136,0.3614,and0.4964 W/m~3,respectively.Ipomoea aquatica had the largest effect on bioelectricity generation promotion.In addition,electrochemically active bacteria,Geobacter and Desulfuromonas,were detected in the anodic biofilm by high-throughput sequencing analysis,and Comamonas(Proteobacteria),which is widely found in MFCs,was also detected in the anodic biofilm.These results confirmed the important role of plants in IVCW-MFCs.     

Identification of potential electrotrophic microbial community in paddy soils by enrichment of microbial electrolysis cell biocathodes

Electrotrophs are microbes that can receive electrons directly from cathode in a microbial electrolysis cell (MEC). They not only participate in organic biosynthesis, but also be crucial in cathode-based bioremediation. However, little is known about the electrotrophic community in paddy soils. Here, the putative electrotrophs were enriched by cathodes of MECs constructed from five paddy soils with various properties using bicarbonate as an electron acceptor, and identified by 16S rRNA-gene based Illumina sequencing. The electrons were gradually consumed on the cathodes, and 25%–45% of which were recovered to reduce bicarbonate to acetic acid during MEC operation. Firmicutes was the dominant bacterial phylum on the cathodes, and Bacillus genus within this phylum was greatly enriched and was the most abundant population among the detected putative electrotrophs for almost all soils. Furthermore, several other members of Firmicutes and Proteobacteria may also participate in electrotrophic process in different soils. Soil pH, amorphous iron and electrical conductivity significantly influenced the putative electrotrophic bacterial community, which explained about 33.5% of the community structural variation. This study provides a basis for understanding the microbial diversity of putative electrotrophs in paddy soils, and highlights the importance of soil properties in shaping the community of putative electrotrophs.     

新型防腐涂层对直升机典型结构材料防护性能研究

目的研究新型防腐涂层对直升机典型结构用铝合金及复合材料在热带海洋性气候环境下的防护性能,并与原防护涂层进行对比。方法通过测试涂层腐蚀后的形貌、光泽度、色差、电化学阻抗等,对比新/原涂层对直升机典型结构用铝合金和复合材料的防护性能。结果经过2年腐蚀后,新涂层表面无明显变化,而原涂层表面有轻微腐蚀,光泽度和色差下降较大。电化学阻抗测试表明,暴露前后涂层结构无明显变化,涂层阻抗值呈现持续下降的趋势,但是原涂层阻抗下降更快。结论新涂层对直升机典型铝合金和复合材料有更好的防护性,更加适于海洋性环境的使用。     

海水中阴极极化对X80钢应力腐蚀及氢脆敏感性的影响

目的研究不同p H值海水中阴极极化对X80管线钢应力腐蚀及氢脆的抑制作用。方法采用慢应变速率拉伸试验、电化学测试、微观组织观察等分析方法。结果 X80钢在天然海水中的析氢电位约为-940m V(vs.SCE,下同),海水p H为3.5时析氢电位发生正移。其应力腐蚀敏感性与极化电位有很大关系,随着极化电位负移,X80钢的氢脆敏感性增加。天然海水中当极化电位负于-950 m V时,断口出现准解理断裂特征形貌。在-1050 m V极化电位下,钢材进入氢脆断裂区发生脆性断裂。海水p H为3.5时,-900 m V钢材有发生氢脆的危险。结论与天然海水相比,X80钢在p H为3.5的酸性海水中具有较高的应力腐蚀敏感性,两种海水介质中X80钢的应力腐蚀敏感性均随极化电位负移而增加。     

7B04铝合金和30CrMnSiA钢短期腐蚀的电化学行为研究

目的研究7B04铝合金和30CrMnSiA合金钢试样的电化学性能。方法采用电化学实验、扫描电子显微镜(SEM)、腐蚀坑深度测试分别对7B04铝合金和30CrMnSiA合金钢试样在是否经过天然海水浸泡的情况下进行测试分析,得到微观腐蚀形貌和腐蚀坑深度以及试样表面不带锈和带锈时的电化学性能。结果随着Na Cl浓度增大,7B04铝合金和30CrMnSiA钢的自腐蚀电流变大,两者之间存在明显的电位差,接触后会发生电偶腐蚀。电偶电流与阴阳极面积比和Na Cl浓度有关,阴阳极面积比增大或Na Cl浓度升高均会导致电偶效应增强。经过海水短期浸泡后,7B04铝合金表面的锈层不仅没有起到抑制腐蚀的作用,反而使自腐蚀加重,但电偶效应确实有所减弱。结论沿海设备在使用过程中应竭力避免铝合金在锈层下工作。     

铜合金实验室试验与实海挂片结果的比较与分析

目的比较实验室试验结果与实海挂片结果对B10和H62两种铜合金腐蚀性能评价的异同之处。方法采用线性极化、电化学阻抗、显微形貌观察等,比较两种铜合金在实验室模拟海水中的的腐蚀速率、点蚀倾向,结合反应机理、腐蚀形貌、环境等外部因素,分析导致实验室与实海环境下出现差异的原因。结果除榆林海域外,实海挂片与实验室结果都表明,B10试样的平均腐蚀速率小于H62。B10和H62两种铜合金随温度升高腐蚀速率增大,在实验室和实海情况下表现出一致性,且平均腐蚀速率都随浸泡时间的延长而降低。结论在实海环境中,温度不是影响点蚀的主要因素,微生物等其他环境因素会对点蚀产生较大影响。