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.     

东南大陆岩石圈板块地体构造

东南大陆的地质构造模型,具有欧亚大陆边缘向洋增生的独特形式。除外来地体之外,或以扩张、地幔上涌,洋壳向大陆地壳转化,并不断向洋增生;或是大陆分裂,地幔柱——热点物质不断注入变薄和破裂了的地壳,形成区域性新的生长构造层,构造岩浆地质体表现为有独特属性的“A型”花岗岩和流纹岩链。这些高DI值的富硅富碱富钠质岩石,出现在欧亚大陆边缘的重力梯度带上。裂解带的地壳结构模式是上地壳存在着重力不稳定的硅铝低速带;中地壳有洋——陆过渡型地壳的“类裂谷型”结构,P波速度为6.3～6.4km/s的中间壳层;在下地壳下部有速度为7.0～7.4km/s的壳——幔混合型高速层;随着“异常”的上地幔的形成,有大范围的热活动和壳——幔边界穹窿,穹窿的地盖比正常区为为薄。由于化学库和化学边界层的横向不连续,可划分出古老基底和后期地质发展史完全迥异的两个亚板块;和以壳层(或幔层)断裂为边界特点的八个地体;一个以分裂为主,并具热点径迹的最新生长构造层的扩张——裂解构造岩浆地体。     

Pearce型构造岩浆地球化学判别图评述——以玄武岩为例

自从六十年代板块构造的概念诞生以来,人们认识到有必要探讨古地理和古构造重建的方法.一段时期以来,很多学者(如Gilluly,1971)认为,岩浆作用与特定的大地构造环境紧密相关,不同的岩浆类型或火山岩系列产在一定的地壳环境,因而出现了许多运用主要或微量元素资料的分类体系或判别图.基于Irvine和Barager(1971)以及Miyashiro(1973)发表的主要元素图解能用来判别岩浆类型,而Pearce(1976)则介绍     

安徽张八岭地区构造演化及动力学

对张八岭地区的构造特征、演化及其动力学有不同解释。通过1：5万张八岭幅、珠龙幅区域地质调查．从地层、岩石组合、岩石化学、岩浆岩、构造变形和变质等研究认为．该区从中新元古代起，经历了成谷、沉积．下沉、顺层剪切、深部熔融、塑性流动、层间褶皱．叠加折劈、宽缓褶曲、逆掩推覆，壳层滑动、熔融成浆、聚浆热隆、侵入或喷发．剥蚀夷平、堆积充填等铸成现今地貌景观．引起上述构造特征和演化的动力源自重力、热力和地球自转速率变化的惯性离心力等的结合与转化，它们在每个阶段各自所起的作用不同，就产生不同构造特征，推动区域构造演化．     

EDTA和腐殖酸对铜在淡水藻(Scenedesmus subspicatus 86.81 SAG)细胞内外分布的影响

讨论了铜在淡水藻（Scenedesmus subspicatus86.81SAG)细胞壁和细胞内分布规律，以及乙二胺四乙酸（EDTA）和水体腐殖酸（FA）存在下对铜分布的影响，研究结果表明，EDTA和FA的存在显著降低了细胞表面铜的吸附量，但是不影响细胞内铜的浓度，对测定数据进行分析。可以认为溶液相铜与EDTA或FA形成的铜络合物是生物无效形态，不参与铜在藻细胞壁上特定位点或生物配体的竞争结合反应。