基于Web的地压实时监测系统

针对矿井井下地压难以有效实时远程监测的现状,开展了井下地压远程监控研究。考虑到井下作业大范围、多参数的复杂性,设计了一个基于串口服务器的Winform与ASP.Net相结合的网络化地压监测系统。对系统结构功能、硬件设计、监测软件设计进行了介绍。现场运行结果验证了该系统的数据传输稳定性、实时性、操作灵活性和扩展性,实现了实时准确地对井下地压的远程监测。     

食蚊鱼(Gambusia afinis)cat、gapdh和gst基因的克隆及其在生态毒理学中的应用

根据Ensembl、Genbank登录的鱼类cat、gapdh和gst基因的CDS序列设计普通PCR扩增引物,寻找食蚊鱼的cat、gapdh和gst基因的c DNA片段,并根据定量引物设计要求设计出相应的SYBR Green I荧光定量RT-q PCR引物,建立了食蚊鱼cat、gapdh和gst基因的SYBR Green I荧光定量RT-q PCR方法。该方法在104~108数量级范围内有良好线性关系(R=0.999~1.000);熔解曲线显示扩增产物特异性良好,均为单一峰值;质粒标准品最高浓度与最低浓度的批内试验变异系数与批间试验变异系数均低于2%。利用该方法监测和评价环境污染物对水生生物的影响,选择了水体中常见典型药物污染物——双氯芬酸,研究其对食蚊鱼抗氧化基因表达的影响。结果表明,雌性食蚊鱼暴露在不同浓度双氯芬酸钠(0.005、0.05、0.5和5 mg·L-1)24 h后,其肝脏cat、gapdh和gst的mRNA呈现显著变化,相对于对照组,在低浓度0.005 mg·L-1时,cat与gst mRNA的表达量均有极显著上升(p0.01),而其它浓度均极显著下降(p0.01)。试验表明该方法具有快速、精确、灵敏度高的优点,可为利用该类小型鱼类的原位污染物的生物监测和生态毒理评价提供有力的技术支持。     

Arsenite transporters expression in rice (Oryza sativa L.) associated with arbuscular mycorrhizal fungi (AMF) colonization under different levels of arsenite stress

As a silicon hyperaccumulator, lowland rice takes up higher levels of As than many other plants due to silicic acid and arsenite sharing the same transporters (Lsi1 and Lsi2). Glomus intraradices (AH01) was inoculated to rice under different arsenite concentrations (0, 2 and 8 μM) in order to investigate the interactions between arbuscular mycorrhizal fungus and rice on the accumulation of arsenite. The relative mRNA expressions of Lsi1 and Lsi2 resulted in a down-regulating trend in mycorrhizal plants. Under 2 μM arsenite treatments, Lsi1 and Lsi2 were significantly decreased, by 0.7-fold (P < 0.05) and 0.5-fold (P < 0.01), respectively, in mycorrhizal plants when compared with non-mycorrhizal plants. This led to the decrease of arsenite uptake per unit of root dry mass. No organic As species were detected in both roots and shoots. The As(III)/As(V) ratios indicated that mycorrhizal plants immobilized most of the arsenite proportion in the roots and prevented its translocation from the roots to the shoots.     

Prenatal and postnatal characterization of Y chromosome structural anomalies by molecular cytogenetic analysis

We describe three cases in which we used fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) and comparative genomic hybridization (CGH) to characterize Y chromosome structural anomalies, unidentifiable by conventional G-banding. Case 1 was a 46,X,+mar karyotype; FISH analysis revealed an entire marker chromosome highlighted after hybridization with the Y chromosome painting probe. The PCR study showed the presence of Y chromosome markers AMG and SY620 and the absence of SY143, SY254 and SY147. CGH results confirmed the loss of Yq11.2-qter. These results indicated the presence of a deletion: del(Y)(q11.2). Case 2 was a 45,X [14]/46,XY[86] karyotype with a very small Y chromosome. The PCR study showed the presence of Y chromosome markers SY620 and AMG, and the absence of SY143, SY254 and SY147. CGH results showed gain of Yq11.2-pter and loss of Yq11.2-q12. These results show the presence of a Yp isodicentric: idic(Y)(q11.2). Case 3 was a 45,X,inv(9)(p11q12)[30]/46,X,idic(Y)(p11.3?),inv(9)(p11q12)[70] karyotype. The FISH signal covered all the abnormal Y chromosome using a Y chromosome paint. The PCR study showed the presence of Y chromosome markers AMG, SY620, SY143, SY254 and SY147. CGH only showed gain of Yq11.2-qter. These results support the presence of an unbalanced (Y;Y) translocation. Our results show that the combined use of molecular and classical cytogenetic methods in clinical diagnosis may allow a better delineation of the chromosome regions implicated in specific clinical disorders. Copyright © 2002 John Wiley & Sons, Ltd.     

Duplex,triplex and quadruplex PCR for the preimplantation genetic diagnosis (PGD) of cystic fibrosis (CF), an exhaustive approach

Most of cystic fibrosis (CF) pre-implantation genetic diagnosis (PGD) cases described to date are limited to the detection of ΔF508. Beside this predominant mutation, over 1000 mutations have been identified, rendering the development of a mutation-based PGD protocol impracticable. This is the reason why we, as well as the others, have developed PGD strategies on the basis of the identification of the pathogenic haplotype instead of the mutation(s). In a previous article, we reported the conditions for the co-amplification of two intragenic polymorphic markers and the F508 locus. Here we describe an improved protocol allowing the additional amplification of two new intragenic markers, intron 1 CA repeat (I1CA) and IVS17bTA. This new protocol should, theoretically, allow us to provide a diagnosis to all couples requiring PGD for CF. Using single lymphoblasts, we have tested four different PCR configurations, including one duplex, two triplexes and one quadruplex PCR. All of them gave results compatible with a clinical application. The number of single lymphoblasts tested in each series varied from 89 to 155. PCR efficiency ranged from 95.4 to 100%. A complete haplotype was achieved for 83.2 to 90.7% of the tested cells, with an allele drop out (ADO) rate comprised between 6.0 and 11.6%. We present here three cases that we performed either with the former test (one case using the triplex PCR combining F508, IVS8CA and IVS17bCA) or with the new one (one case using the triplex combining F508, I1CA and IVS17bTA and one case using a quadruplex test). We obtained two single pregnancies. Copyright © 2004 John Wiley & Sons, Ltd.     

分子生物学方法在微生物多样性及微生物生态研究中的应用

首先介绍了分子生物学技术在微生物多样性及微生物生态研究中应用的理论基础，以及在此基础上引入的分子生物学技术如：PCR、DNA杂交技术在分子层面上来研究微生物遗传多样性，并结合已有工作积累展望了分子微生物生态研究的发展前景．图2参20