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.     

农闲期城郊农田灌溉水中典型污染物含量与评价——以西北江三角洲为例

摸查了农田灌溉系统水环境经农（夏）闲期降水等外部强干预调理后的环境污染物含量——该值理论上是环境自净作用之后的最不利值,对农田灌溉系统水环境进行水质评价,探究其对农田土壤质量的潜在污染风险.于2019年6月中旬—7月下旬（降雨集中期）,分别在西北江三角洲城市（清远市、佛山市和江门市）实验基地周边筛选研究区,并在雨后对有覆水的水源区域、灌渠、蓄水池和田间水等采集上覆水,共采得水样27×2份,对其pH值、悬浮物（SS）、矿化度、总磷（TP）、氨氮（NH₄⁺-N）、Cd、As、Pb、Cu和Zn的含量进行检测;对各研究区pH值、SS的成因和影响,矿化度的等级,重金属均值分布等进行分析;对全样品TP、NH₄⁺-N、重金属含量进行Pearson相关性分析和描述性特征分析;通过单因子水质标识指数P_i和综合水质标识指数P分别对各采样点、各构成项目和各研究区进行水质评价.研究发现雨后农田灌溉系统水环境构成复杂,物质呈无显著性差异的迁移,灌溉沿程上覆水中大部分物质可能处于动态平衡,受局部环境影响变小,TP、NH₄⁺-N和重金属等迁移物质主要还是灌溉水体中原有的,受外力驱动扰动后在水动力作用下可能以氮磷结合形态沿灌溉系统发生远距离迁移;使用河流水质标识指数法评价农田灌溉系统水环境,发现该法可以刻画局部灌溉的水质态势,对农田灌溉系统水环境的含量特征评价做出科学、合理的解释,也可以做出综合性定量评价.虽然灌溉水系统结构差异较大,但是从整体上可以初步得出农闲期西北江三角洲农田灌溉水的灌溉风险不高的结论,总体综合水质评价级别为Ⅰ类~Ⅱ类,达到水环境功能区的使用要求.     

电絮凝净化含聚污水影响因素研究

目的为保障油田正常安全生产,促进油田含聚污水的循环利用。方法采用电絮凝技术单因素研究方法开展净化含聚污水的静态实验研究,探讨外加电流密度、极板间距、初始pH及聚合物浓度等因素对净化效果的影响规律。结果当初始pH为7.0,电流密度为4.0 m A/m2,极板间距为1.0 cm,电解16 min时,综合处理效果最佳。此时,含油量与浊度去除率分别为98.85%,99.93%,处理每克油平均消耗为0.0494 g的Al,处理每立方米含聚污水的能耗为0.2895 k Wh。污水中的聚合物浓度越小,净化效果越好。结论中性处理,极板间距为1.0 cm,外加电流密度为4.0 m A/m2,通电16 min是最佳处理条件。     

预应力管桩的工作机理及其在基坑支护中的应用

分析了预应力管桩受力破坏的机理和单桩承载力提高的原因,以及预应力管桩作为基坑支护桩的受力特性,并以武汉南国置业有限公司高层公寓综合楼为工程实例,进一步阐明采用预应力管桩作为基坑竖向加固桩,具有明显的经济效益。     

4-氯苯酚在水溶液中的光化学反应(Ⅰ)单线态氧和自由基的产生

在模拟太阳光照射下，４氯苯酚（４ＣＰ）浓度迅速降低，反应过程对应着产生大量的单线态氧和自由基．金属离子Ｆｅ２＋、Ｆｅ３＋、Ａｌ３＋能够加速这一过程，尤以Ｆｅ２＋、Ｆｅ３＋的影响更明显；在模拟太阳光照射下，向４ＣＰ体系加入的富里酸（ＦＡ），对４ＣＰ浓度降低略起抑制作用，这表明二者之间存在相互作用．但当再加入金属离子Ｆｅ２＋、Ｆｅ３＋、Ａｌ３＋后，能够加速４ＣＰ浓度降低，尤以Ｆｅ２＋、Ｆｅ３＋的影响更显著．     

土壤生态系统微量元素的生物有效性研究现状

土壤中植物必需的微量营养元素的供给以及有害元素的毒性大小并不仅仅与元素的总量有关,而取决于元素的有效性。文章论述了元素生物有效性的含义,介绍了元素有效性的研究方法,即连续偏提取法和单提取法,并对一些相关问题进行了深入的分析讨论。     

单细胞凝胶电泳技术在环境污染物检测中的应用

主要介绍了单细胞凝胶电泳技术(SCGE),包括SCGE的原理、方法及其在环境污染物检测中的应用,该技术可用多种污染物的检测且灵敏度高,具有广阔的应用前景。即SCGE对大气污染物、重金属、醛类及辐射污染的检测,具有高度敏感性,是具有广阔应用前景的新技术。     

滑坡灾害危险性评价模型比较

通过对庆元地区滑坡灾害的详细研究，比较讨论了单变量评价、决策树评价与逻辑回归法等3种滑坡危险性评价模型在实际应用中的效果。单变量模型最为简单，计算方便，得到的结果也具有较高的准确性，当收集到的滑坡因素越多，对滑坡发育的刻画越精确，则最终评价结果也越可信；决策树算法能迅速从大量样本中归纳出一般规则，并具有较高的精度，但其结果也受到训练样本数量大小和样本维数即参与评价的因素集数量的影响；逻辑回归模型具有计算方法简单、评价结果物理意义明确等突出优点，但需要有大量样本数据的支持才能取得较好的效果。     

菲和镉单一及复合污染条件下在毛蚶体内的富集动力学研究

采用室内半静态双箱动力学模型实验,研究了菲和镉单一及复合污染条件下在毛蚶(Anadara subcrenata)体内的生物富集,通过对富集与释放过程中毛蚶体内菲和Cd的富集量进行非线性曲线拟合,获得了菲和Cd单一及复合污染条件下在毛蚶体内的吸收速率常数k1、释放速率常数k2、生物富集因子BCF、生物半衰期B1/2和平衡状态下最大富集量CA max等动力学参数。实验结果表明,菲和Cd在实验前期富集速率较高,8 d以后富集速率减缓,释放阶段与富集阶段相似。毛蚶对菲的BCF值为37.80,远大于Cd的BCF值13.12,且生物半衰期时间更长,菲更容易在生物体内富集。菲和Cd联合暴露条件下,在毛蚶体内的CA max和BCF值均大于单一作用,说明二者同时暴露时,毛蚶对菲和镉的吸收富集均有所增强。实验模型拟合度较好,输出值和实测值之间无显著性差异,拟合方程和拟合参数可信。