微流控芯片快速鉴定多重细菌

建立了一种用于多重细菌鉴定的微流控芯片分析方法。在芯片上实现细菌进样、培养和鉴定,结合培养池阵列的空间分辨力以及菌种特异性显色培养基的颜色分辨力,可以实现多重细菌检测。实验选用4种泌尿系统感染常见病原菌作为模拟测试对象,结果显示,这种芯片方法在15 h内可完成细菌鉴定,检测限可达101cfu/mL。临床样本测试结果显示,芯片方法可以实现4种细菌同步鉴定,检测结果与传统方法的一致性达到96.3%。这种微流控细菌鉴定方法简便快速,有望发展成为细菌检测的有力工具。     

开放式微流控芯片上的肿瘤组织微阵列构建

构建了一种具有自动形成细胞培养阵列和多步骤灵活操作特点的开放式微流控芯片。此芯片具有3层复合式结构:底层为微通道贯穿的细胞培养池阵列,顶层是开放式培养池,二者之间为一层纳米孔薄膜。纳米孔薄膜具有"透气阻水"的特性,既起到止流阀作用实现液体自动分配,又允许跨膜扩散,模拟血管内皮层扩散屏障结构。结合移液工作站,开放式微流控芯片可以完成细胞换液、药物处理和细胞活力测试等一系列分析步骤。本研究以乳腺癌细胞为模型,在芯片上90 s内可构建包含三维细胞培养和仿真血管内皮的10×10肿瘤组织微阵列。形态学和免疫组织化学检测证实了芯片肿瘤组织的仿真效果。抗肿瘤药物测试结果表明,这种开放式微流控组织阵列芯片允许在仿真条件下进行包含复杂操作步骤的细胞实验,是细胞研究的有利工具。     

阵列微流控浓度梯度网络用于细胞-化学刺激反应研究

设计和制作了具有5组平行浓度梯度形成网络和30个细胞培养池的微流控芯片,该芯片集成了细胞接种、培养、梯度浓度化学刺激、标记和检测等功能单元。芯片为玻璃-PDMS杂合结构,微流控通道刻蚀于玻璃层。芯片细胞培养池设计了系列围堰结构以利于细胞贴壁。细胞接种、灌流培养和试剂引入通过外接微量注射泵控制完成。该芯片可以生成连续、稳定的平行浓度梯度。观察发现,围堰结构有利于细胞接种和生长,乳腺癌MCF-7细胞在芯片灌流培养条件下生长良好。利用该芯片检测了在接受As2O3和乙酰丝氨酸(NAC)梯度浓度刺激后乳腺癌MCF-7细胞内谷胱甘肽(GSH)水平以及细胞阿霉素敏感性的变化,分析乳腺癌细胞阿霉素敏感性与细胞内GSH水平的关系。MCF-7细胞内GSH水平的变化与刺激药物浓度呈剂量-效应依赖关系,在接受As2O3刺激后GSH水平有所下降;而在接受NAC刺激后GSH水平有所升高。MCF-7细胞阿霉素敏感性与GSH水平相关。在降低GSH水平后药物敏感性升高;而升高细胞内GSH水平后敏感性降低。这种阵列微流控浓度梯度网络可以用于高通量细胞-化学刺激反应研究,有潜力成为细胞水平大规模药物筛选的技术平台。     

微流控细胞芯片LED诱导荧光检测微系统

基于微流控细胞芯片分析技术和微机电系统(MEMS)加工技术, 设计制作了阵列式微流控细胞检测芯片, 采用自组装的顶窗型光电倍增管(PMT)和信号采集电路采集芯片微管道内流动细胞的荧光信号, 构建了一种针对低浓度细胞悬浮液的微流控细胞芯片发光二极管(LED)诱导荧光的快速检测微系统, 实现了对低浓度(≤40 Cell/mL)荧光标记的HepG2肝癌细胞悬浮液样本的定量计数和测试, 而且在血液细胞共存的条件下, 仍可以有效地对血液中少量HepG2肝癌细胞进行荧光计数和测试. 整个系统结构简单, 操作方便且检测灵敏度较高.     

基于液芯波导原理的微流控芯片长光程光度检测系统

提出了一种基于液芯波导(Liquidcorewaveguide,LCW)原理的微流控芯片吸收光度检测系统.通过芯片与外界接口技术实现液芯波导管与芯片的耦合,建立了芯片上长光程(毫米至厘米级)吸收光度检测池.采用邻菲啉-铁(Ⅱ)显色体系验证系统分析性能,以5.5cm外覆TeflonAF液芯波导管作为检测池(检测池体积240nL)时,芯片系统的检测线性范围为0.03～50μmol/L,对邻菲啉-铁(Ⅱ)配合物的检出限为8nmol/L,检测池有效光程达1.7cm,分析精度RSD(n=5)为0.8%.     

基于微流控芯片的细菌趋化性检测研究进展

细菌趋化性是指有运动能力的细菌对环境化学物质梯度产生响应,趋向某些化学诱导剂或避开某些化学驱避剂的移动行为,是微生物适应环境变化而生存的一种基本属性.研究细菌趋化性对于利用细菌治理环境、控制病原菌侵染机体以及开发微生物工业项目等方面都具有重要意义.微流控芯片可以实现对细菌趋化性的定性与定量检测,与传统的检测方法相比,可以更好地对细菌的微环境进行控制,有较高的灵敏度.近年来,基于微流控技术检测细菌趋化性研究得到了飞速发展.本文从微流控芯片的结构、工作方式及主要应用3个方面对近年出现的微流控趋化性检测装置进行了介绍和评述.     

阵列纸芯片比色法检测碱性磷酸酶

采用5-溴-4-氯-3-吲哚磷酸盐(BCIP)/氯化硝基四氮唑蓝(NBT)显色体系,构建了阵列纸芯片比色检测碱性磷酸酶(ALP)的方法.首先,借助烘干处理方式在光刻法制备的阵列纸芯片微孔中固定显色试剂,然后加入ALP进行显色反应,最后,采用凝胶成像仪和普通照相机成像,读取显色强度(灰度值)进行比色检测.详细考察了显色条件对检测结果的影响,探讨了人血清白蛋白对ALP检测的增色效应,在最佳实验条件下,ALP检测的线性范围为1.5～20 U/L,检出限(3 σ)为0.78 U/L(n=18),比文献报道中纸芯片上检测ALP方法的检出限低约两个数量级.本方法成功用于实际血清样品检测,测定结果与临床值一致.在此基础上,构建了双色阵列纸芯片,通过颜色的变化实现了ALP的可视化半定量检测.     

基于离心式微流控技术的乙肝病毒检测新方法

基于传统的酶联免疫吸附测定(ELISA)原理,提出了一种基于离心式微流控技术的乙肝病毒检测新方法。设计制作的一次性芯片集成进样、酶联免疫反应、显色反应及检测单元。选用乙肝表面抗原作为实验对象,在芯片上提前包被检测抗体,分三个阶段完成待检样品中乙肝表面抗原的检测。将芯片上测试结果使用Origin软件拟合,相关系数R~2达到0.99212,芯片能够在60 min内实现对乙肝病毒表面抗原的快速精准检测,对乙肝表面抗原加标回收率为95%~102.3%,制作的芯片在一个月内的稳定性好。与传统乙肝检测方法比较,离心式微流控芯片检测方法具有检测速度快、稳定性好、检测线性好和操作简单等优点。     

用于细胞三维培养的集成微柱阵列的微流控芯片设计与验证

设计并验证了一种用于细胞三维培养的集成微柱阵列的微流控芯片.芯片由一片聚二甲基硅氧烷(PDMS)沟道片和一片玻璃盖片组成, 在PDMS沟道片上集成了一个由两排微柱阵列围成的细胞培养室和两条用于输送培养基的侧沟道.微柱间距直接影响了芯片的使用性能, 是整个芯片设计的关键.基于数值模拟和实验验证, 本研究对微柱间距进行了优化设计.优化后的微流控芯片可以很好地实现细胞与细胞外基质模拟材料混合液的稳定注入、培养基中营养物质向培养室内的快速扩散和细胞代谢物的及时排出.在芯片上进行了神经干细胞的三维培养, 证明了芯片上构建的细胞体外微环境的稳定性.     

异硫氰酸荧光素标记万古霉素的制备及其与细菌相互作用的荧光检测

以异硫氰酸荧光素（FITC）标记万古霉素（Vanco）制备了FITC Vanco（FV）,FV经高效液相制备色谱纯化后,进行了质谱结构鉴定。 分别通过荧光分光光度计和流式细胞仪（FCM）测定不同浓度FV培养基中培养细菌细胞的荧光强度。 结果显示,所制备的荧光探针对5种细菌有特异性结合,用荧光强度可表征细胞结合抗生素的数量,对细菌耐药机制研究和细菌对万古霉素作用敏感性的测试有应用价值。     

二氧化碳和丙烯直接合成甲基丙烯酸的NiPMo/TiO2催化剂的研究

用溶胶-凝胶法以磷钼酸(MPA)的镍盐溶液水解钛酸四丁酯制备了NiPMo/TiO2催化剂.使用ICP、 XRD、 TG-DTA、 IR、 TPD-MS和微反应技术研究了催化剂的化学组成、热稳定性、化学吸附性质和催化反应性能.杂多钼酸盐与TiO2通过O2-在TiO2表面发生了键合.在623 K下,杂多阴离子仍保持原有的Keggin结构.CO2在Lewis酸位Ni(Ⅱ)和Lewis碱位Ni-O-Mo的桥氧协同作用下生成CO2卧式吸附态Ni(Ⅱ)←O-(CO)←(O--Ni).丙烯有多种吸附态在催化剂上吸附.在563 K、 1 MPa和空速1500 h-1的反应条件下,丙烯的摩尔转化率为3.2%,产物MAA选择性为95%.     

Synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives

Different approaches for the synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives from simple amino acids have been investigated. A library of 33 precursors for the preparation of N-hydroxy pyrazinones was obtained in moderate to good yields.     

A general and convenient synthesis of 4-(tosylmethyl)semicarbazones and their use in amidoalkylation of hydrogen,heteroatom, and carbon nucleophiles

A general synthesis of previously unknown semicarbazone-based α-amidoalkylating reagents, 4-(tosylmethyl)semicarbazones, has been developed. The synthesis involved three-component condensation of semicarbazones of aliphatic or aromatic aldehydes with the same or other aldehydes and p-toluenesulfinic acid. The scope and limitations of this reaction were investigated. The compounds obtained were demonstrated to be an efficient α-(4-semicarbazono)alkylating agents. They were reacted with H- (sodium borohydride), O- (sodium methylate), S- (sodium phenylthiolate), N- (pyrrolidine, sodium succinimide), P- (trialkyl phosphites), and C-nucleophiles (sodium diethyl malonate) to give the corresponding products of the tosyl group substitution, 4-substituted semicarbazones, including analogues of nitrofurazone. Among the prepared compounds tested in vitro for antibacterial and antifungal activity, three nitrofuryl-containing semicarbazones exhibited high biological activities with minimum inhibitory concentration (MIC) values of 8–32 μg/mL.     

Toward new camptothecins. Part 6: Synthesis of crucial ketones and their use in Friedländer reaction

In the context of the preparation of camptothecin and luotonin A analogs, the synthesis of some key keto-precursors and their use in Friedländer condensation are described. This paper also focuses on the stability of these keto intermediates and emphasizes the major differences between indolizinones and pyrroloquinazolinones series. Noteworthy is also the report of some original structures isolated as by-products of some experiments.     

Synthesis of novel chiral bidentate hydroxyalkyl-N-heterocyclic carbene ligands and their application in palladium-catalyzed Mizoroki–Heck couplings and asymmetric addition of diethylzinc to benzaldehyde

A small library of new chiral bidentate hydroxyalkyl-imidazolium salts 1 is conveniently synthesized on multi-gram scale from inexpensive and commercially available chiral pool amino acids. The corresponding carbenes, generated by deprotonation of imidazolium salts 1, in combination with palladium(II) chloride were tested in the Mizoroki–Heck coupling reaction. The most significant results in terms of yields and reactivities were achieved with low catalyst loading. The catalytic activities of these imidazolium salts were also investigated in the asymmetric addition of diethylzinc to benzaldehyde. The use of MgO nanoparticles as an additive in conjunction with these ligands played a crucial role in increasing the efficiency of these reactions.     

α-Amino acid derived enaminones and their application in the synthesis of N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates and other heterocycles

A new and simple synthesis of novel N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates, which exist in equilibrium with their 4-oxo tautomers, has been developed in two steps starting from N-protected α-amino acids. The key intermediates are enaminones, which can also be isolated, characterized, and used for the construction of other functionalized heterocycles, before they spontaneously decompose to pyrrole products. 4-Hydroxypyrroles are prone to partial aerial oxidation but can be efficiently alkylated or reduced to stable polysubstituted pyrrolidine derivatives.     

Specific intramolecular aromatic CH insertion of diazosulfonamides

The chemoselectivity in the intramolecular CH insertion of various diazosulfonamides has been experimentally studied. The results reveal that the aliphatic 1,4-, 1,5-, or 1,6-C(sp³)?H insertions of diazosulfonamides are not accessible, while the aromatic 1,5-C(sp²)?H insertion can be realized specifically by adjusting the diazo-adjacent group. In addition, the general chemoselectivities in the intramolecular CH insertions of diazosulfonyl compounds are summarized. Generally, diazosulfones undergo both aromatic 1,5-C(sp²)?H and aliphatic 1,5- and 1,6-C(sp³)?H insertions, while diazosulfonates undergo aliphatic 1,5- and 1,6-C(sp³)?H insertions. However, diazosulfonamides only undergo aromatic 1,5-C(sp²)?H insertion.     

CoFe2O4自形成磁性液体场致结构化对磁化的影响

The Langevin paramagnetic theory can’t describe the relation between magnetization of ferrofluids and applied magnetic field. The structuralization of ferrofluids, which is considered the main influence factor of the magnetization, is regarded. The part of magnetization works is deposited when the structure is forming. This action influences the magnetization of ferrofluids directly or indirectly. On the base of the “compressing” model, the Langevin function that usually describes the magnetization of ferrofluid is modified, and a well-fitted curve is obtained. An equation of the relation between the equivalent volume fraction after being “compressed” and the intensity of magnetic field is discovered, which approximately describes the process of magnetization. The relation between the approximate initial susceptibility and the volume fraction can be obtained from modified formula.     

Microwave-assisted copper-catalyzed stereoselective ring expansion of three-membered heterocycles with α-diazo-β-dicarbonyl compounds

Microwave-assisted copper-catalyzed ring expansions of three-membered heterocycles with α-diazo-β-dicarbonyl compounds were investigated. Thiiranes generated 3-acyl-5,6-dihydro-1,4-oxathiines in the presence of copper sulfate and trans-3-acyl-5,6-dihydro-1,4-oxathiines as stereospecific products for 1,2-disubstituted cis-thiiranes through an intramolecular S_N2 process. Oxiranes gave rise to 2-acyl-5,6-dihydro-1,4-dioxines under the catalysis of copper hexafluoroacetylacetonate and cis-3-acyl-5,6-dihydro-1,4-dioxines as stereospecific products for 1,2-disubstituted cis-oxiranes via an intimate ion-pair mechanism. The current method provides a direct and simple strategy in efficient preparation of 3-acyl-5,6-dihydro-1,4-oxathiines and 2-acyl-5,6-dihydro-1,4-dioxines, important agents in medicinal and agricultural chemistry, from readily available thiiranes and oxiranes, respectively.     

KMnO4-mediated oxidative CN bond cleavage of tertiary amines: Synthesis of amides and sulfonamides

KMnO₄-mediated oxidative CN bond cleavage of tertiary amines producing secondary amine was introduced, which was trapped by electrophiles (acyl chloride and sulfonyl chloride) to form amides and sulfonamides. The reaction could take place at mild condition, tolerating a wide range of function groups and affording products in moderate to excellent yields.