First-trimester prenatal diagnosis of spinal muscular atrophy using microsatellite markers

Twenty-five pregnancies at risk for spinal muscular atrophy I (SMA I) have been monitored by first-trimester prenatal diagnosis. Microsatellite markers were used in all cases to amplify polymorphic regions at the D5S125, D5S435, D5S39, D5S127, and D5S112 loci. All families, including 12 SMA I pedigrees with a deceased index child, were fully informative for DNA analysis. Three fetuses were predicted to be affected and 22 fetuses were predicted to be unaffected. Twenty-two newborns were unaffected by clinical examination at birth. These results support the accuracy of SMA I prenatal diagnosis based on linkage analysis.     

Maternal contamination of amniotic fluid demonstrated by DNA analysis

DNA from 16 sets of samples comprising DNA from uncultured amniotic fluid cells, cultured amniotic fluid cells, fetal tissue, and maternal blood was analysed by the polymerase chain reaction (PCR) with AC-repeat primers. The analysis was performed to investigate the presence of contaminating maternal cells in amniotic fluid which would affect the reliability of DNA studies for prenatal diagnosis. In three sets, maternal contamination of uncultured amniotic fluid cells was detected. In one of the three sets, maternal contamination was present in both uncultured and cultured amniotic fluid cells. The use of amniotic fluid cells as a source of DNA for prenatal diagnosis should be limited to cases where the purity of the DNA can be demonstrated prior to the diagnostic test being performed. This limitation in the use of amniotic fluid DNA also extends to other forms of diagnosis relying on the purity of amniotic fluid samples, particularly the new in situ hybridization methods currently being developed.     

基于模糊综合评判的电网应灾能力量化评估*

为应对极端自然灾害下电网在灾前、灾后应灾能力评估的不足,提出基于模糊综合评判的电网应灾能力量化评估方法。通过分析电网系统的综合数据和关联指标,建立电网灾前承受力和灾后恢复力评估指标体系;运用层次分析法和熵权法各自求得指标主、客观权重,并依据权重占比组合成综合权重,使用模糊综合评判对电网应灾能力进行量化评估;通过案例对所提的评估进行验证。结果表明:该评估方法更加有效体现电网实际应灾能力水平,对电网应对极端自然灾害具有一定指导作用。     

富勒烯对PCR反应的抑制作用

以110bp单链DNA为模板,研究富勒烯(C60)对聚合酶链式反应(polymerase chain reaction,PCR)的影响.实验结果表明,随着C60浓度的增加,PCR反应被显著抑制;将Taq DNA聚合酶、单链DNA模板与C60孵育后,其PCR扩增产物均显著减少;增加PCR反应体系中的Taq DNA聚合酶量,可消除C60的抑制作用,但增加起始单链DNA模板的数量,效应不明显,上述研究结果说明,C60不仅可抑制Taq DNA聚合酶活性,同时对DNA模板也具有一定的损伤作用.     

Effects of p,p'-DDE exposure on gonadal development and gene expression in Japanese medaka (Oryzias latipes)

Although 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p'-DDE),the major and most persistent metabolite of dichlorodiphenyl-trichloroethane (DDT),was continually detected in wild fishes that showed abnormal gonad development such as intersex,little is known about the impact of p,p'-DDE exposure on gonad development in fishes.To survey the effects of p,p'-DDE on gonadal development and gene expressions,male juvenile (20-d post hatch) Japanese medaka (Oryzias latipes) was exposed to 1,5,20,and 100 μg/L p,p'-DDE for two months.Increased hepatosomatic index (HSI) and decreased gonadosomatic index (GSI) were found in the p,p'-DDE-treated groups.Intersex was found in 100 μg/L p,p'-DDE exposure group,as well as 100 ng/L 17α-ethynylestradiol (EE2)group.By quantitative real-time RT-PCR,it was found that gene expressions of vitellogenins (VTG-1,VTG-2),choriogenins (CHG-H,CHG-L),and estrogen receptor α (ER-α) in the liver of the fish were significantly up-regulated by p,p'-DDE exposure.VTC-1 and VTG-2 were recommended as the preferred biomarker for assessing anti-androgenic p,p'-DDE because they were the highest up-regulated among the genes and showed good dose-response relationship.The up-regulated ER-α suggested that a potential synergetic effect would occur when p,p'-DDE coexists with other ER-α-binding endocrine-disrupting chemicals (EDCs).     

膜生物反应器对白斑综合症病毒的去除研究

研究了不同孔径的膜生物反应器对游离白斑综合症病毒(white spot syndrome virus,WSSV)的截留效果,对不同膜生物反应器的进水、上清液、出水进行荧光定量PCR检测.结果表明,0.45 μm孔径的膜组件对磷酸缓冲液中WSSV的去除量仅0.6 lg;0.22 μm孔径的膜对WSSV病毒的去除量为1.18 lg;0.1 μm孔径的膜对WSSV的去除量为5.5 lg.进一步研究表明,在养殖废水中添加游离的WSSV时,随着时间的延长,膜组件的生物样物质对WSSV去除起了重要的作用;孔径为0.45 μm的膜生物反应器,对WSSV的去除量也达到5.35 lg;3种不同孔径膜生物反应器中运行12h后,对WSSV的去除率并没有显著差异.     

水中轮状病毒实时定量PCR外标准品的构建

采用细胞培养和T-A克隆技术,在轮状病毒主要结构蛋白VP7基因序列上设计合成引物,经PCR扩增后将特异性产物连接入pGEM-T-easy载体中,经过酶切鉴定和测序分析获得轮状病毒cDNA标准品.利用常规PCR和实时定量PCR方法对所获得的cDNA标准品进行特异性、稳定性和重复性指标的检验.结果表明,利用此标准品制备的标准曲线具有较高的扩增效率和良好的线性关系(斜率为-3.353,R2=0.995);实时定量PCR熔解曲线分析表明,温度在81℃±0.5℃的PCR产物是轮状病毒VP7序列上的特异性产物,表明此标准品是轮状病毒特异性的;同时,所制备的标准品在实时定量PCR检测中具有较大的线性范围(9×100～9×1011 copies/μL,每个反应最低可以检测到9个拷贝数的轮状病毒cDNA,表明利用该标准品进行实时定量PCR分析时具有较高的检测灵敏性;此外,该标准品具有较高的稳定性和重复性(3次独立实验CV值在0.2%～0.9%之间),可长期稳定保存.构建的标准品可用作检测水环境中轮状病毒的实时荧光定量PCR的外标准品.     

Ag-AgIO3改性BiOI光催化剂湿法脱除气态单质汞

采用沉积沉淀-光还原法制备了不同AgIO₃含量的BiOI光催化剂,利用X射线衍射、N₂吸附-脱附、扫描电子显微镜、X射线光电子能谱、紫外-可见漫反射光谱和电子自旋共振对其物理化学结构进行表征,在光催化反应器上研究了AgIO₃负载量、光照、溶液温度、pH值、SO₂和NO对气态单质汞（Hg⁰）去除性能的影响.结果表明,与BiOI相比,Ag-AgIO₃改性后光催化剂的脱汞性能大幅提高.当AgIO₃负载量为4wt.%时,光催化剂的脱汞效率高达98%.与NO相比,SO₂对脱汞性能的抑制作用更大.由于Ag和AgIO₃在BiOI表面的高度分散性,Ag-AgIO₃（4%） BiOI的可见光吸收性能明显提高.荧光灯辐照与Ag-AgIO₃/BiOI光催化剂协同会产生大量的活性物种.在Ag-AgIO₃/BiOI光催化剂的高效脱汞体系中,超氧阴离子自由基（·O₂^-）为最主要的活性物质,而空穴（h⁺）和羟基自由基（·OH）次之.     

活性污泥中的腐殖酸对实时荧光定量PCR的影响

活性污泥基因组中的腐殖酸会影响实时荧光定量PCR检测的准确性.本研究采用抽提基因组前腐殖酸去除试剂盒、抽提基因组后腐殖酸去除试剂盒、抽提基因组前后共同去除试剂盒3种不同的方法,去除活性污泥基因组中的腐殖酸,经过PCR和实时荧光定量PCR方法的验证找出腐殖酸去除的最适方法;通过DNA酶Ⅰ消化基因组DNA保留杂质腐殖酸,向其中掺入已知浓度的总细菌质粒,5个浓度10倍梯度稀释10 ng·μL~(-1)的定量模板,探究腐殖酸和梯度稀释模板对活性污泥总细菌实时荧光定量PCR的影响.结果显示,在不同处理组中,通过反复洗脱的方式在基因组抽提前去除活性污泥中的腐殖酸,DNA的得率(162.46 ng·μL~(-1))最多,A260/230值为1.34,A260/280值为1.80,腐殖酸的残留最少;去除腐殖酸后外加内标质粒组,梯度稀释10 ng·μL~(-1)模板100倍及以上,腐殖酸的抑制效应显著降低后保持不变(p0.05),100倍以内变化不显著(p0.05);抽提前去除腐殖酸组进行实时荧光定量PCR的抑制效率为6.16%,显著低于对照组的72.68%(p0.05).研究表明,抽提前去除活性污泥腐殖酸,抽提后将基因组模板稀释100倍可显著降低活性污泥实时荧光定量PCR的抑制效率,提高定量结果的准确性.     

西江流域地表水微生物污染定量源解析

水体微生物污染标记物的源强特征是定量解析微生物污染来源的关键.应用实时荧光定量PCR（qPCR）技术分别对人源（qHS601F/qBac725R）、牛源（BacB2-590F/Bac708Rm）、猪源（Bac41F/Bac163R）及禽类（qC160F-HU/qBac265R-HU）特异性拟杆菌引物的源强特征进行了探索,并进一步对西江流域广东段进行微生物污染来源定量解析,结果表明：针对不同目标宿主,不同特异性拟杆菌引物的源强特征为人源（qHB） > 猪源（qPB） > 牛源（qRB） > 禽类（qCB）,DNA模板浓度对引物的源强特征影响较小.选取的目标研究区域水体普遍受到人、猪、牛及禽类粪便污染.干流中qHB、qCB及qPB自上游至下游缓慢升高.各支流中qHB、qCB及qPB的平均浓度较干流均上升了1个数量级,qRB在不同河段变化较小.说明下游河段的生活污水、猪源及禽类粪便污染较为严重,牛源污染的影响较小.统计分析显示在污染水平较高的各支流中,人源特异性拟杆菌引物与传统指示菌具有一定的相关性（P<0.05）,说明生活污水是传统指示菌浓度较高的主要原因,且为持续排放源.