New technique using galactose-specific lectin for isolation of fetal cells from maternal blood

To isolate fetal cells from maternal blood, we developed a new method based on galactose-bearing conjugation. Nucleated red blood cells (NRBCs), which highly express galactose on their surface, were selectively attached to a substrate coated with a galactose-containing polymer via soybean agglutinin (SBA), a galactose-specific lectin. Cord blood samples were used to evaluate enrichment efficacy of NRBCs by this method. Blood samples were obtained from 131 pregnant women between 6 and 27 gestational weeks. After preliminary condensation of fetal cells by Ficoll gradient centrifugation, NRBCs were enriched using galactose-positive selection by adjusting SBA concentration. We isolated one to severalhundred NRBCs (mean±SD, 7.8±8.5) in 2.3 ml of peripheral blood samples from 96% of pregnantwomen. The isolated NRBCs were analyzed by a Y-chromosome FISH probe in eight cases carrying male fetuses. Y-signals were detected in all eight cases and more than half of the NRBCs wereoffetal origin. The study demonstrates that our new method using galactose-specific lectin provides effective enrichment of fetal NRBCs allowing non-invasive prenatal diagnosis. Copyright © 2002 John Wiley & Sons, Ltd.     

用高浓度对苯二甲酸溶液产电的微生物燃料电池

以高浓度对苯二甲酸（TA）溶液为底物,研究微生物燃料电池的产电效果.以厌氧活性污泥作为接种体,经过210 h驯化,开路电压达到0.54 V,证明了TA可以作为微生物燃料电池的底物进行产电.深入研究了不同pH值和底物浓度对产电的影响,实验结果表明,当体系pH为8.0时,负载两端（R=1 000 Ω）电压最大,底物浓度越高,负载两端电压越大,并逐渐趋近于一个最大值,通过Monod方程回归得到该微生物燃料电池体系输出电压的最大值U_max为0.5 V,K_s值为785.2 mg/L.当底物浓度（以COD计）为4 000 mg/L时,最大输出功率密度为96.3 mW/m²,库仑效率为2.66%,COD去除率为80.3%.     

剩余污泥燃料电池处理含铬废水的效能及机理

采用剩余污泥为阳极底物,六价铬为阴极电子受体,构建双室微生物燃料电池(MFC).MFC启动成功后,考察阳极室污泥初始浓度和阴极室六价铬初始浓度对MFC产电性能及六价铬还原速率的影响.较高的污泥浓度(8~12g/L)对六价铬的还原速率影响均较小,且去除率均可达99%以上.污泥浓度为10g/L的MFC具有较高的产电性能,内阻为108Ω,最大功率密度输出为3621mW/m3.阴极室较高的Cr(VI)初始浓度可维持较长时间的高输出电压,但对阳极污泥降解并无明显影响.XPS测试结果表明,阴极Cr(VI)的还原产物为Cr(III),因电场作用被吸附在电极片上,使得阴极溶液中的总铬浓度降低.研究表明,剩余污泥为底物的微生物燃料电池可以在产电的同时实现剩余污泥的资源化及电镀废水的无害化.     

电催化-生物电化学耦合系统处理青霉素废水的机制

抗生素生产过程中产生大量含有残存抗生素的生产废水,传统的污水生物处理技术难以有效地处理此类高浓度抗生素废水.针对此问题,采用电催化-生物电化学耦合系统来处理含有典型的β-内酰胺类抗生素青霉素的废水,利用硼掺杂金刚石（boron-doped diamond,BDD）电催化电极对青霉素废水进行预处理,其出水进入生物电化学系统（bioelectrochemical system,BES）进行后处理.研究发现,经电催化系统预处理后青霉素的去除率为89%,出水进入BES后可以稳定运行,该出水中又有79%的青霉素被BES去除,获得最大功率密度为（1124±28）mW·m^-2,与直接进青霉素原始废水的BES反应器相比提高了473%.经过电催化-生物电化学两级耦合系统处理后青霉素的总去除率达到98%.对BES反应器阳极生物量和生物相分析结果表明,青霉素对阳极混合菌群生物量和变形菌门微生物（主要产电菌）有一定的抑制作用,且会降低形成阳极生物膜的主要微生物不动杆菌属Acinetobacter和具有产电功能芽孢杆菌属Bacillus在反应器中含量,这是影响反应器产电性能和处理效果的主要原因.青霉素废水经电催化降解后,浓度明显降低,有效缓解了青霉素对BES的抑制作用,提高废水的可生化性,因此电催化-生物电化学耦合系统是一种高效低能耗处理抗生素废水的新工艺.     

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.     

石墨烯掺杂聚苯胺阳极提高微生物燃料电池性能

微生物燃料电池（microbial fuel cell,MFC）技术可分解代谢污染物质并同步输出电能,在环境及能源领域吸引了越来越多的关注.但是,输出功率密度较低、成本较高、底物降解率低等特点限制了其实际应用,其中阳极是主要限制因素之一.本研究选取具有优异导电性、大比表面积的石墨烯和生物相容性较好的聚苯胺（polyaniline,PANI）,并优化二者比例关系,制备得到石墨烯掺杂PANI复合材料.将复合材料涂覆在玻碳电极表面分析电化学性能,循环伏安（cyclic voltammetry,CV）和线性伏安扫描（linear sweep voltammetry,LSV）测试结果均显示石墨烯含量占比20%的复合电极（20%石墨烯）电化学性能最好.将复合材料修饰在碳布表面作为MFC阳极时以石墨烯含量占比5%的复合电极（5%石墨烯）生物电化学性能最佳,LSV得到最大输出功率密度为（831±45）mW·m^-2,分别是20%石墨烯、1%石墨烯、石墨烯、PANI、碳布阳极的1.2、1.3、1.3、1.5、1.8倍.最大输出电压、开路电压、化学需氧量去除率、库仑效率、生物量密度均以5%石墨烯电极最高.电化学阻抗分析表明5%石墨烯电极极化内阻仅为（24±2）Ω,是碳布电极的19.8%.电化学和生物电化学性能并不完全一致,说明电极材料的生物相容性是影响MFC性能的主要因素之一.5%石墨烯阳极充分发挥了石墨烯和聚苯胺的优点,提高了MFC的产电性能.     

Prenatal diagnosis of triple X using fetal cells obtained by endocervical lavage

Prenatal diagnosis of aneuploidies currently relies on invasive procedures such as chorionic villous sampling and amniocentesis. These methods can provide an accurate diagnosis of the fetal karyotype but are relatively invasive. Here, we report a case of the prenatal detection of triple X using fetal cells obtained by lavage of the endocervical canal prior to amniocentesis, following ultrasound diagnosis of hydrops fetalis. Copyright © 2003 John Wiley & Sons, Ltd.     

双室微生物燃料电池处含银废水的产电性研究

以剩余污泥为阳极底,糖蜜废水为基质,Ag Cl废水为阴极电子受体,构建了双室微生物燃料电池(Two-chamber Microbial Fuel Cell,简称MFCs),并研究了电池的产电特性、库仑效率及金属去除率.结果表明:Ag+不仅可以作为阴极电子受体,而且还能稳定产电,外电阻为1000Ω时,获得的最大电压为514.5 m V,最大功率密度为65.82 m W·m-2.在阴极实现了对废水中Ag+的去除,最大去除率可达71.6%,而且Ag+浓度为2000 mg·L-1时,回收金属银单质质量为197.66 g.在阳极对废水的处理效果也很显著,库仑效率最高为2.66%,COD去除率最大为81.22%.     

Synergistic cytotoxicity of binary combinations of inorganic and organic disinfection byproducts assessed by real-time cell analysis

Chlorine, chlorine dioxide, and ozone are widely used as disinfectants in drinking water treatments. However, the combined use of different disinfectants can result in the formation of various organic and inorganic disinfection byproducts (DBPs). The toxic interactions, including synergism, addition, and antagonism, among the complex DBPs are still unclear. In this study, we established and verified a real-time cell analysis (RTCA) method for cytotoxicity measurement on Chinese hamster ovary (CHO) cell. Using this convenient and accurate method, we assessed the cytotoxicity of a series of binary combinations consisting of one of the 3 inorganic DBPs (chlorite, chlorate, and bromate) and one of the 32 regulated and emerging organic DBPs. The combination index (CI) of each combination was calculated and evaluated by isobolographic analysis to reflect the toxic interactions. The results confirmed the synergistic effect on cytotoxicity in the binary combinations consisting of chlorite and one of the 5 organic DBPs (2 iodinated DBPs (I-DBPs) and 3 brominated DBPs (Br-DBPs)), chlorate and one of the 4 organic DBPs (3 aromatic DBPs and dibromoacetonitrile), and bromate and one of the 3 organic DBPs (2 I-DBPs and dibromoacetic acid). The possible synergism mechanism of organic DBPs on the inorganic ones may be attributed to the influence of organic DBPs on cell membrane and cell antioxidant system. This study revealed the toxic interactions among organic and inorganic DBPs, and emphasized the latent adverse outcomes in the combined use of different disinfectants.     

四环素类抗生素对淡水绿藻的毒性作用

四环素类抗生素是目前世界上应用最为广泛的抗生素之一,也是我国畜禽养殖业中使用量最大的兽药和饲料添加剂.由于该类物质不易被生物体吸收且当前的污水处理设施对抗生素不能彻底去除,故大量抗生素及其代谢产物最终进入地表水环境,其对水生态系统和人体健康的潜在威胁值得关注.本研究以水生态系统初级生产者淡水绿藻为受试生物,系统考察四环素、金霉素和强力霉素对蛋白核小球藻、斜生栅藻细胞膜通透性和生长抑制作用的影响.结果表明,抗生素暴露96 h后,强力霉素在整个作用浓度范围内使供试淡水绿藻细胞膜通透性降低;而金霉素和四环素低浓度暴露可以增大藻细胞膜通透性,此后随着作用浓度的增大转为降低.3种抗生素对淡水绿藻生长抑制率大致呈现强力霉素>四环素>金霉素的顺序趋势.斜生栅藻对四环素类抗生素的毒性响应较蛋白核小球藻更为敏感.     

利用分子生物等技术确定活性污泥硝化细菌的衰减特征

通过常规耗氧速率（OUR）测定方法、荧光原位杂交技术（FISH）和LIVE/DEAD细胞活性实验,分别研究了序批式反应器（SBR）硝化系统和生物营养物去除（BNR）系统中硝化细菌的好氧衰减特征.实验结果表明,SBR硝化系统中硝化细菌在衰减过程中由细胞死亡引起的衰减分别占细胞总衰减的33%（SRT=10 d）和50%（SRT=40 d）;相应地,由活性降低引起的衰减分别占细胞总衰减的67%（SRT=10 d）和50%（SRT=40 d）.长SRT可能会选择出能够更好地适应饥饿状态的硝化菌种,使细菌能够迅速地做出紧迫反应,从而降低其衰减速率.在BNR系统中（SRT=15 d）,硝化细菌在衰减过程中由细胞死亡引起的衰减约占细胞总衰减的45%,由活性降低引起的衰减约占细胞总衰减的55%.两种系统中硝化细菌由细胞死亡引起的衰减比例不同是由于两种系统中不同微生物组成所致.     

聚磷菌富集实验及其内源特征探究

通过磷酸盐释放速率(PRR)测定、荧光原位杂交技术(FISH)及LIVE/DEAD细胞染色技术,分别研究了生物营养物去除(BNR)系统与富集聚磷菌(PAOs)序批式反应器(SBR)系统中PAOs在好氧环境下的衰减特征.结果表明,当富集聚磷菌SBR系统进料中碳源(三水合乙酸钠和丙酸)是以36d为一个循环周期方式投加时,即三水合乙酸钠24d和丙酸12d,系统中PAOs富集比例可达91%.测定与计算结果表明,生物营养物去除(BNR)系统与富集聚磷菌SBR系统中PAOs衰减速率分别为0.113d-1和0.181d-1,死亡速率分别为0.048d-1和0.036d-1.这说明由细胞死亡引起的PAOs数量衰减在两个系统中分别占细胞总衰减的42%(BNR)和20%(SBR),而由细胞活性降低引起的活性衰减分别占细胞总衰减的58%(BNR)和80%(SBR).由此可见,PAOs数量衰减在其细胞总衰减中只占较小一部分,而绝大部分衰减是由活性衰减所引起.     

仿生脂肪细胞制备以及对水体中林丹去除的研究

利用生物体脂肪组织可以对脂溶性有机物有效富集的原理,通过界面聚合合成了类似脂肪细胞结构的仿生脂肪细胞,并对其结构进行了初步表征.仿生脂肪细胞具有亲水性膜以及亲脂性的内部结构,亲水性的膜允许携带脂溶性有机物的水体穿过膜,亲脂性的内含物将脂溶性有机物富集截留.仿生脂肪细胞对林丹具有较好的去除效果,15%三油酸甘油酯含量的仿生脂肪细胞与粉末活性炭具有相当的林丹去除能力.仿生脂肪细胞对林丹的去除机理包括内含物的生物富集以及膜上空腔的物理吸附两部分,而内含物的生物富集作用则是主要作用方式.     

微藻型微生物燃料电池的研究进展

在石油资源日趋紧张以及环境恶化日趋严重的今天,微生物燃料电池(MFC)因其可同时实现污水处理和能源回收而受到广泛关注。微藻技术与MFC技术结合产生的微藻型MFC系统得到证实并随之兴起,其中尤以微藻生物阴极型MFC因可实现污水处理、零碳排放、CO2捕捉、太阳能捕获及电能、生物柴油、藻体残渣有价回收等多重功能,成为研究热点。文章根据其中微藻所起的不同作用将微藻型MFC系统分成三类,在参阅大量文献的基础上进行了全面综述,并由此对构建高效微藻生物阴极型MFC进行了探讨,提出了计算机辅助菌种选择技术等相关设想,最后对微藻型MFC的发展提出了展望。     

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。     

羧基化碳纳米管载铂催化剂对微生物燃料电池阴极氧还原性能的影响

阴极催化剂是影响微生物燃料电池(microbial fuel cell,MFC)性能的关键要素.为了考察不同羧基化方法改性的碳纳米管(carbon nanotube,CNT)负载Pt的催化氧还原效率,分别在80℃和95℃条件下对CNT进行了羧基化,采用浸渍-沉淀法制备了Pt/CNT催化剂(Pt/CNT-80和Pt/CNT-95),并在空气阴极MFC体系中验证了其催化氧还原效果(MFC-80、MFC-95和MFC-C).结果表明,MFC-95和MFC-80的最大功率密度分别为568.8 mW.m-2和412.8 mW.m-2,内阻分别为204.7Ω和207.7Ω,开路电压分别为0.719 V和0.651 V.而对照MFC-C的最大功率密度仅为5.4 mW.m-2,内阻为826.2Ω.XPS和XRD分析结果显示,Pt/CNT-95催化氧还原效果优于Pt/CNT-80,原因可能是95℃羧基化过程在CNT表面引入了丰富的含氧基团.     

葡萄糖和硝基苯为混合燃料时MFC的产电特性研究

通过构建双极室微生物燃料电池(microbial fuel cell,MFC),以铁氰化钾溶液为阴极电子受体,以硝基苯(nitrobenzene,NB)和葡萄糖为混合燃料,研究MFC的产电特性和NB的降解情况.结果表明,在外阻为1000Ω的条件下,随着NB初始浓度的增加,双极室MFC的产电特性明显受到抑制.当葡萄糖浓度为1000mg/L,NB初始浓度分别为0、50、150、250mg/L时,MFC的运行周期逐渐缩短,分别为55.7、51.6、45.9、32.2h;最大输出电压分别为670、597、507、489mV;最大体积功率密度分别为28.57、20.42、9.29、8.47W/m3;电荷量分别为65.10、43.50、35.48、30.32C.MFC利用NB和葡萄糖为混合燃料,可以在稳定地输出电能的同时实现有机物高效降解,MFC对NB去除率高达100%,对COD的去除率达到87%～98%.但以250mg/LNB为单一燃料时,MFC无明显产电现象.DGGE图谱表明NB的加入改变了MFC阳极电极上微生物的群落结构.     

双酚A体外类雌激素活性评价及其作用机制的初步研究

采用体外MCF-7细胞增殖试验检测了双酚A(BPA)的类雌激素活性，并用生长曲线分析、细胞周期分析、他奠昔芬拮抗试验和凋亡检测对其作用机制进行了初步探讨。结果显示BPA有明显刺激MCF-7细胞增殖的类雌激素活性；细胞周期分析BPA组细胞增殖指数均明显高于溶剂对照组；BPA刺激MCF-7细胞增殖的类雌激素活性可被Tam拮抗：BPA能明显抑制MCF-7细胞凋亡的发生。结果提示BPA刺激MCF-7细胞增殖的类雌激素活性作用机制可能是BPA与雌激素受体结合后，影响细胞增殖及细胞凋亡过程，从而产生一系列生物效应。     

Magnetic-activated cell sorting (MACS) significantly decreases the hybridization efficiency of fluorescence in situ hybridization (FISH)

Fetal cells were enriched from maternal blood using density gradient centrifugation of Histopaque followed by magnetic-activated cell sorting (MACS) to select CD71-positive cells. For each specimen, cells partially purified by Histopaque were split into equal portions, and each portion was subjected to purification by MACS in parallel. Cells before and after MACS were subjected to dual-color fluorescence in situ hybridization (FISH) analysis with X- and Y-chromosome-specific probes. We found that the hybridization rates were decreased by approximately 10% after MACS based on duplicated analysis for each sample. Copyright © 2001 John Wiley & Sons, Ltd.     

好氧生物阴极型微生物燃料电池的同时硝化和产电的研究

在两室型微生物燃料电池的阴极室接种硝化菌实现了同时硝化和产电.硝化过程和产电过程在同一区域实现,不仅能够充分利用曝气的溶解氧,节省曝气能源消耗,而且硝化过程产生的额外的质子,有效地避免了产电过程所造成的阴极pH值升高.运行稳定期间MFC的最大电流和最大功率密度分别为47mA和45.50W/m3,当进水氨氮浓度为153.4mg/L时,硝化速率为5.98mg/(L·d).硝化菌会与产电菌竞争溶解氧,但当溶解氧浓度控制在3.5～5.0mg/L时,硝化过程未对产电产生明显影响.无缓冲溶液的条件下,加入氨氮时的阴极电势比未加入氨氮时的阴极电势高124mV,且阴极电势变化的阶段与氨氮降解的过程是一一对应的.H+离子的理论计算表明,硝化过程产生的H+离子(8.14×10-3mol)与产电过程消耗H+离子(8.54×10-3mol)数量相当,证实了硝化作用中产生的H+离子能够补偿阴极室由于产电造成的H+离子的消耗,维持系统pH值的稳定.