Experience in quantitative analysis of floristic structure and diversity in Samarskaya Luka

Quantitative methods have been applied to the study of floristic structure and diversity in Samarskaya Luka. The results show that Samarskaya Luka is a floristically heterogeneous area in which six elementary floras can be distinguished. This area has been assessed as a floristically autonomous formation, and its floristic representativeness was estimated.     

First-trimester fetal sex determination in maternal serum using real-time PCR

An Erratum has been published for this article in Prenatal Diagnosis 22(13) 2002, 1241. Fetal sex prediction can be achieved using PCR targeted at the SRY gene by analysing cell-free fetal DNA in maternal serum. Unfortunately, the results reported to date show a lack of sensitivity, especially during the first trimester of pregnancy. Therefore, determination of fetal sex by maternal serum analysis could not replace karyotype analysis following chorionic villus sampling. A new highly sensitive real-time PCR was developped to detect an SRY gene sequence in maternal serum. Analysis was performed on 121 pregnant women during the first trimester of pregnancy (mean gestational age: 11.8 weeks). Among them, 51 had at least one previous male-bearing pregnancy. Results were compared with fetal sex. SRY PCR analysis of maternal serum was in complete concordance with fetal sex. Among the 121 pregnant women, 61 were bearing a male fetus and 60 a female fetus. No false-negative results were observed. Furthermore, no false-positive results occurred, even though 27 women carrying a female fetus during the current pregnancy had at least one previous male-bearing pregnancy. This study demonstrates that a reliable, non-invasive sex determination can be achieved by PCR analysis of maternal serum during the first trimester of pregnancy. This non-invasive approach for fetal sex prediction should have great implications in the management of pregnant women who are carriers of an X-linked genetic disorder. Prenatal diagnosis might thus be performed for male fetuses only, avoiding invasive procedures and the risk of the loss of female fetuses. Copyright © 2001 John Wiley & Sons, Ltd.     

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.     

现行财税体制下我国耕地资源数量变化的实证研究

耕地资源数量变化受到人口、经济等诸多因素的影响，而财税体制是影响耕地资源数量变化的更深屡次原因。通过数量关系的初步观察和作用机理分析说明我国现行财税体制对耕地资源数量变化存在正反两方面作用，总体来看，对耕地资源保护不利。计量结果进一步证实了我国现行财税体制对耕地资源数量变化存在显著影响。揭示了其影响的方向和大小。根据研究结果，文章有针对性地提出了增加地方财政预算收入，合理分配土地出让金比例和科学使用土地出让金等相关政策建议。     

Pitfalls in molecular diagnosis in a family with severe factor VII (FVII) deficiency—misdiagnosis by direct sequence analysis using a PCR product

Molecular diagnostic tests are becoming a routine analysis in many laboratories. These modern analyses are widely used in clinical medicine, forensic, genetic and prenatal diagnosis and also in preimplantation genetic diagnosis. The accuracy of analysis is highly dependent on the success achieved in minimising genotyping errors. The pitfalls in molecular diagnostic tests can be due to a simple technique such as the polymerase chain reaction (PCR) used universally. This technique is routinely used for its apparent accuracy, but it is also a well-known source of errors. We report an error introduced during PCR reaction that leads to a wrong sequence result and consequently to a ‘false’ molecular result in a next prenatal diagnosis in a family with severe factor VII (FVII) deficiency. This error was verified using an unsuitable primer design in a rich repetitive sequence of the FVII gene that leads to a false annealing and then to a wrong molecular diagnosis. It is essential to link closely molecular data with clinical and phenotype analysis in order to avoid false-negative or false-positive results, which is of great importance to diagnosis and molecular prevention. Copyright © 2003 John Wiley & Sons, Ltd.     

pH值对亚硝酸盐氧化菌动力学及功能基因的影响

为探究pH值对亚硝酸盐氧化菌（NOB）活性动力学影响,本试验采用序批式活性污泥（SBR）反应器,以富含NOB的活性污泥为对象,基于Monod模型考察不同pH值对NOB活性动力学的影响并进行统计学分析.结果表明,Monod方程可较好地反映不同pH值条件下基质底物浓度对NOB比亚硝态氮氧化速率（SNiOR）的影响,且pH=7.0时动力学参数Ks为（6.167mg/L）,r_max为[1.134g/（g·d）],此时NOB活性最好.利用钟形经验模型进行非线性回归拟合,最大比降解速率（r_max）随pH值的增大呈钟形变化,本试验NOB的最佳pH值为（6.9±0.1）,其中r_max维持在r_opt一半以上的pH值范围（ω）为（3.26±0.4）.以亚硝酸盐氧化还原酶类基因（nxrA、nxrB）为引物,基于荧光定量PCR技术分析结果显示,在不同pH值条件下nxrA基因和nxrB基因拷贝数的变化趋势均与动力学参数（Ks、r_max）的规律一致,且nxrA和nxrB基因在系统的降解过程中起协同作用.     

红树林土壤解磷菌的分离鉴定及解磷特性

从罗源湾红树林根际土壤中分离解磷菌,筛选出洋葱伯克霍尔德菌（Burkholderia cepacia, NR 113645.1）和短小芽孢杆菌（Bacillus pumilus, NR 043242.1）,研究它们的解磷特性和动态解磷过程. HPLC结果表明, B. cepacia菌液上清中含葡萄糖酸、丙酮酸、乳酸、乙酸、丁二酸, 这些有机酸的产生导致磷酸盐溶解. B.pumilus菌液上清中仅检测到少量的葡萄糖酸, 因此该菌溶磷效果不佳. 在细菌动态溶磷过程中发现, 经B. cepacia菌处理的磷酸钙, 其X射线衍射（XRD）特征峰强度随时间的增长降低, 显微镜图像显示磷酸钙颗粒随时间增长变小, 甚至消失. 而经B.pumilus菌处理的结果表明, 在试验期间内磷酸钙峰强度和颗粒大小没有明显变化. 细菌产生有机酸是溶解磷酸盐的重要前提, 这是解决板结土壤中难溶性磷源转化为生物可利用磷源问题的关键.     

中国“四化”建设对水足迹强度的影响分析

采用2000—2016年中国30省市（不包括香港、澳门、台湾、西藏四省区）的数据,构建四化指标体系,计算四化发展指数,进行地区四化水平评价,通过OLS和GMM回归模型考察四化发展对水足迹强度的影响,同时设置四化之间的交互项来识别四化指标之间的相互作用。主要结论如下：首先,从四化水平的地域发展差异上来看,中国四化水平总体上呈现出东部优于中部、中部优于西部的格局,水足迹强度的分布与之类似,说明四化水平发展与水足迹强度之间可能存在相关关系;其次,计量模型显示四化水平的提升对于水足迹强度的改善有积极影响,但每一化对于水足迹强度改善的影响大小不一;第三,以交互项衡量的四化相互作用对于水足迹强度的影响作用为正,表明四化发展对水足迹强度的改善效果逐渐趋于收敛。因此,优先提升落后地区的四化水平,以四化水平的提升推动水足迹强度的降低,能够有效降低我国总体水足迹强度水平。