Ratiometric Detection of microRNA Using Hybridization Chain Reaction and Fluorogenic Silver Nanoclusters

miRNA (miR)-155 is a potential biomarker for breast cancers. We aimed at developing a nanosensor for miR-155 detection by integrating hybridization chain reaction (HCR) and silver nanoclusters (AgNCs). HCR serves as an enzyme-free and isothermal amplification method, whereas AgNCs provide a built-in fluorogenic detection probe that could simplify the downstream analysis. The two components were integrated by adding a nucleation sequence of AgNCs to the hairpin of HCR. The working principle was based on the influence of microenvironment towards the hosted AgNCs, whereby unfolding of hairpin upon HCR has manipulated the distance between the hosted AgNCs and cytosine-rich toehold region of hairpin. As such, the dominant emission of AgNCs changed from red to yellow in the absence and presence of miR-155, enabling a ratiometric measurement of miR with high sensitivity. The limit of detection (LOD) of our HCR-AgNCs nanosensor is 1.13 fM in buffered solution. We have also tested the assay in diluted serum samples, with comparable LOD of 1.58 fM obtained. This shows the great promise of our HCR-AgNCs nanosensor for clinical application.     

A paper-based resonance energy transfer nucleic acid hybridization assay using upconversion nanoparticles as donors and quantum dots as acceptors

Monodisperse aqueous upconverting nanoparticles (UCNPs) were covalently immobilized on aldehyde modified cellulose paper via reduction amination to develop a luminescence resonance energy transfer (LRET)-based nucleic acid hybridization assay. This first account of covalent immobilization of UCNPs on paper for a bioassay reports an optically responsive method that is sensitive, reproducible and robust. The immobilized UCNPs were decorated with oligonucleotide probes to capture HPRT1 housekeeping gene fragments, which in turn brought reporter conjugated quantum dots (QDs) in close proximity to the UCNPs for LRET. This sandwich assay could detect unlabeled oligonucleotide target, and had a limit of detection of 13 fmol and a dynamic range spanning nearly 3 orders of magnitude. The use of QDs, which are excellent LRET acceptors, demonstrated improved sensitivity, limit of detection, dynamic range and selectivity compared to similar assays that have used molecular fluorophores as acceptors. The selectivity of the assay was attributed to the decoration of the QDs with polyethylene glycol to eliminate non-specific adsorption. The kinetics of hybridization were determined to be diffusion limited and full signal development occurred within 3 min.     

New organoruthenium metallates containing ferrocenecarboxalidine thiosemicarbazones and their nucleic acid/albumin binding and in vitro cytotoxicity

A string of four new hetero binuclear Ru(III) complexes of ferrocenecarboxaldehyde-4(N)-substituted thiosemicarbazones were synthesized and characterized by various spectral (infrared, ultraviolet–visible, Electron Paramagnetic Resonance (EPR) and High Resolution Mass Spectrometry (HR-MS) techniques. The binding abilities of the ligands/complexes with nucleic acid (calf thymus DNA, CT-DNA) and bovine serum albumin (BSA) were analyzed by absorption and emission titration methods. The complexes exhibited better DNA binding affinity than their parent ligands. The interaction with CT-DNA was found to be intercalative and with BSA static quenching mechanism was observed. All the synthesized Ru(III) complexes were subjected to study their in vitro cytotoxicity against MCF-7 (human breast cancer) and HT-29 (human colon cancer) cell lines. Among the four complexes, complex 3 [RuCp (FF-etsc)PPh₃]Cl exhibited the highest cytotoxicity in MCF-7 cells and complex 4 [RuCp (FF-ptsc)PPh₃]Cl was the most active on HT-29 cells.     

Micro enzymatic assay coupled to capillary electrophoresis via liquid junction

Summary A system for capillary electrophoresis combined with enzymatic assay has been evaluated for the two enzymes glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. Instrumentation included a post-column reactor coupled to the separation capillary by a liquid junction. A technique for generating a substrate solution flow into the reactor by utilizing two high voltage supplies is proposed. This method offers a high degree of freedom in optimizing the separation and enzymatic reaction conditions individually. Possibilities for improving the enzymatic assay sensitivity were also examined.     

Energy of formation for and liquid binary alloys

We have investigated the free energy of formation for Ag_xIn_1-x and Ag_xSn_1-x liquid binary alloys at temperatures 1173 and 1250 K, respectively. A microscopic theory based on the first order perturbation has been applied. The interionic interaction and a reference liquid are the fundamental components of the theory. The interionic interaction is described by a local pseudopotential. A liquid of hard spheres (HS) of two different effective diametres and charges is used to describe the reference system. The results of the calculations for energy of formation agree very well with the available experimental data. Our calculations also reveal that a simple perturbative approach along with appropriate effective pair potentials can produce nearly quantitative results for the concerned alloys.     

一个在水稻胚胎中差异表达基因的分离鉴定

通过筛选水稻开花后48～50h的原胚和120～122h的分化胚的cDNA文库,得到一个在胚胎发育过程中表达有差异的克隆,命名为OsEES,测序结果显示OsEES含有一个预测的亚精氨合成酶结构域,编码一个大小约38×103(pI6.5)的蛋白.同时RNA原位杂交的结果显示开花48～50h的原胚中的信号强度明显大于开花120～122h的分化胚,表明OsEES可能在水稻胚胎发生过程中起到一定的调控作用.     

Quantum Hall effect in back-gated InAs/GaSb heterostructures under a tilted magnetic field

We investigate the quantum Hall effect (QHE) in the InAs/GaSb hybridized electron–hole system grown on a conductive InAs substrate which act as a back-gate. In these samples, the electron density is constant and the hole density is controlled by the gate-voltage. Under a magnetic field perpendicular to the sample plane, the QHE appears along integer Landau-level (LL) filling factors of the net-carriers, where the net-carrier density is the difference between the electron and hole densities. In addition, longitudinal resistance maxima corresponding to the crossing of the extended states of the original electron and hole LLs make the QHE regions along integer-ν_net discontinuous. Under tilted magnetic fields, these R_xx maxima disappear in the high magnetic field region. The results show that the in-plane magnetic field component enhances the electron–hole hybridization and the formation of minigaps at LL crossings.     

槲寄生蛋白注射液细胞毒活性测定的MTT方法探讨

采用MTT比色法对体外培养的肿瘤细胞进行细胞毒作用实验，验证槲寄生蛋白注射液体外抗肿瘤效果．并且鉴定这种检测方法的有效性。实验结果表明槲寄生蛋白注射液有一定的体外抗肿瘤效果；MTT比色法是一种可用的体外细胞毒作用检测法，为新药品的开发提供了实验依据。