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MicroRNAs (miRNAs) play important regulatory roles in a wide range of biological processes, and their aberrant expression is associated with cancer development and a variety of diseases. Here, we develop a simple, sensitive, and specific miRNA assay on the basis of circular exponential amplification in combination with the hairpin probes. The binding of target miRNA with a linear DNA template initiates the first strand displacement amplification (SDA) and generates the universal triggers which are complementary to the 3' protruding end of a hairpin probe. These universal triggers function not only as the primers to unfold the hairpin probes through an extension reaction, generating distinct fluorescence signals, but also as the amplification templates to initiate the second SDA reaction. Moreover, the second SDA reaction can release new triggers to initiate the above two consecutive SDA reactions, thus constituting a circular exponential amplification which enables the conversion of a small amount of miRNAs to a large number of universal triggers to unfold abundant hairpin probes. This hairpin probe-based circular exponential amplification assay exhibits high sensitivity with a detection limit of 3.80 × 10-13 M and a detection range of 4 orders of magnitude. It can even discriminate single-nucleotide difference between miRNA family members and perform well in real sample analysis. Notably, in this assay, the long-stem hairpin probes are unfolded through an extension reaction rather than through a conventional hybridization reaction controlled by the thermodynamic equilibrium in the case of molecular beacons, making the design of hairpin probes very simple. This hairpin probe-based circular exponential amplification assay holds a great promise for further application in biomedical research and early clinical diagnosis. [ABSTRACT FROM AUTHOR]

Sensitive and specific detection of DNA methylation in CpG sites of genomic DNA is imperative for rapid epigenetic evaluation and early cancer diagnosis. Here, we employ for the first time the thermostable ligation for methylated DNA discrimination and hyperbranched rolling circle amplification (HRCA) for signal enhancement, without the need for restriction enzymes, PCR amplification, or fluorescence-labeled probes. After bisulfite treatment of methylated DNA, the methylation-specific linear padlock probe can be circularized only in the presence of methylated DNA and serves subsequently as a template for HRCA, whose products are easily detected using SYBR Green I and a standard fluorometer. While in the presence of unmethylated DNA, the linear padlock probe cannot be circularized because of the defectively matched substrate, and no HRCA occurs. This ligation-mediated HRCA-based method exhibits excellent specificity and high sensitivity with a detection limit of 0.8 fM and a detection range of 4 orders of magnitude, and it can even distinguish as low as 0.01% methylation level from the mixture, which is superior to most currently used methods for DNA methylation assay. This method can be further applied to analyze genomic DNA in human lung cancer cells. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are important regulators of gene expression that control almost every physiological and pathological process. Although the complementarity between the seed region of a miRNA and its target mRNA is usually deemed as the key determinant in the miRNA-target recognition in animals, the mechanism of their recognition still remains enigmatic as more and more exceptions challenge the seed rule. Herein, we employ single-molecule fluorescence resonance energy transfer (smFRET) to investigate human miRNA-induced silencing complex (miRISC)-involved miRNA-target recognition with either perfect base pairing or poor seed match in real time. Our results demonstrate that the recognition between mammalian mIRNA and its target with perfect base pairing proceeds in a two-state model as prokaryotic guide DNA-mediated recognition, suggesting a conserved pattern of guide RNA/DNA strand recognition. In addition to the general rule of mIRNA-target recognition, our results reveal that annealing between miRNA and its target with poor seed match proceeds in a stepwise way, which is in accordance with the increase in the number of conformational states of miRNA-target duplex accommodated by the miRJSC, suggesting the structural plasticity of human miRISC to conciliate the mismatches m seed region. This new dynamic information revealed by smPRET has an important implication for comprehensive understanding of the role of m1RJSC in the target recognition in mammals. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are associated with various diseases. Here we develop a rapid, highly sensitive, and specific miRNA assay based on the two-stage exponential amplification reaction (EXPAR) and a single-quantum-dot (QD)-based nanosensor. The two-stage EXPAR involves two templates and two-stage amplification reactions under isothermal conditions. The first template enables the amplification of miRNA, and the second template enables the conversion of miRNA to the reporter oligonucleotide. Importantly, different miRNAs can be converted to the same reporter oligonucleotides, which can hybridize with the same set of capture and reporter probes to form sandwich hybrids. These sandwich hybrids can be assembled on the surface of 605 nm emission QDs (605QDs) to form the 605QD/reporter oligonucleotide/Cy5 complexes, where the 605QD functions as both a fluorescence resonance energy transfer donor and a target concentrator. Upon excitation with a wavelength of 488 nm, distinct Cy5 signals can be observed in the presence of target miRNA. This assay is highly sensitive and specific with a detection limit of 0.1 aM and can even discriminate single-nucleotide differences between miRNA family members. Moreover, in combination with the specific templates, this method can be applied for multiplex miRNA assay by simply using the same set of capture and reporter probes. This highly sensitive and specific assay has potential to become a promising miRNA quantification method in biomedical research and clinical diagnosis. [ABSTRACT FROM AUTHOR]

Recent advances in single-molecule detection, nanotechnology, and aptameric sensors hold exciting promise for many potential applications. By functionalizing the surface of a quantum dot (QD) with aptamers which can recognize cocaine, and taking advantage of single-molecule detection and fluorescence resonance energy transfer (FRET) between 605QD and Cy5 and Iowa Black RQ, we develop a single-QD-based aptameric sensor that is capable of sensing the presence of cocaine through both signal-off and signal-on modes. In comparison with the established aptameric sensors, this single-QD-based aptameric sensor has the significant advantages of simple sample preparation, high sensitivity, and extremely low sample consumption. With the advances in the development of varieties of aptamers for small molecules, nucleic acids, metal ions, and proteins, this single-QD-based aptameric sensor might find wide application in forensic analysis, environmental monitoring, and clinic diagnostics. [ABSTRACT FROM AUTHOR]

The sequence-specific RRE RNA-Rev binding is essential for HIV-1 replication and provides a useful in vitro system for real-time evaluating the inhibitory effect of drugs on the RRE-Rev interaction. The rapid and sensitive detection of RRE-Rev interaction in complex biological systems represents a fundamental challenge. Here we report the development of a single-quantum-dot (QD)-based nanosensor for sensitively quantifying Rev peptide-RRE interaction and characterizing the potential inhibitors by virtue of single-molecule detection and QD-based fluorescence resonance energy transfer (FRET). We demonstrate that the stoichiometry of Rev peptide binding to RRE can be accurately determined by using this single-QD-based nanosensor. Importantly, this single-QD-based nanosensor can sensitively quantify the inhibitory efficacy of proflavin on the Rev peptide-RRE binding, even in the presence of substantial levels of interference fluorescence from high-concentration proflavin, which usually prevents the discrimination of FRET signals in ensemble measurements. The application of this nanosensor in the screening of libraries of small-molecule drugs will facilitate the development of new drugs against various diseases, cancers, and HIV. [ABSTRACT FROM AUTHOR]

Fluorescence resonance energy transfer (FRET)-based nanosensors with quantum dots (QDs) as donors and organic dyes as acceptors have long been of interest for the detection of biomolecules such as nucleic acids, but their low FRET efficiency in bulk solution has prevented the sensitive detection of nucleic acids due to the large size of the QDs and the long length of nucleic acids. Here we describe a novel approach to improve the detection sensitivity of QD-based nanosensors using single-molecule detection in a capillary flow. In comparison with bulk measurement, single-molecule detection in a capillary flow possesses the unique advantages of improved FRET efficiency, high sensitivity, prevention of photobleaching, and low sample consumption. Greater FRET efficiency was obtained due to the deformation of DNA in the capillary stream. This technique can be easily extended to sensitive bimolecular analysis in microfluidic chips, and it may also offer a promising approach to study the deformation of small nucleic acids in fluid flow. [ABSTRACT FROM AUTHOR]

Rapid and highly sensitive detection of DNA is critical in diagnosing genetic diseases. Conventional approaches often rely on cumbersome, semi-quantitative amplification of target DNA to improve detection sensitivity. In addition, most DNA detection systems (microarrays, for example), regardless of their need for target amplification, require separation of unhybridized DNA strands from hybridized stands immobilized on a solid substrate, and are thereby complicated by solution–surface binding kinetics. Here, we report an ultrasensitive nanosensor based on fluorescence resonance energy transfer (FRET) capable of detecting low concentrations of DNA in a separation-free format. This system uses quantum dots (QDs) linked to DNA probes to capture DNA targets. The target strand binds to a dye-labelled reporter strand thus forming a FRET donor–acceptor ensemble. The QD also functions as a concentrator that amplifies the target signal by confining several targets in a nanoscale domain. Unbound nanosensors produce near-zero background fluorescence, but on binding to even a small amount of target DNA (∼50 copies or less) they generate a very distinct FRET signal. A nanosensor-based oligonucleotide ligation assay has been demonstrated to successfully detect a point mutation typical of some ovarian tumours in clinical samples. [ABSTRACT FROM AUTHOR]

Aim of Study: Colorectal cancer (CRC), now the third most common cancer across the world, is known to aggregate in families. Recently, genome-wide association studies have identified two single nucleotide polymorphisms (SNP) associated with CRC in Caucasians. Materials and Methods: To validate whether the same variations conferred risk to CRC in the Han Chinese population, we genotyped 760 individuals (380 controls and 380 cases samples) recruited from the Han Chinese origin. Results: We found rs11987193 in 8p12 (P = 0.0472 after correction, OR = 0.751) was significantly associated with CRC but rs12080929 in 1p33 (P = 0.0650 after correction, OR = 0.750) was not. Conclusion: Our findings supported that rs11987193 is a susceptibility locus for CRC, and gene DUSP4 was possible to play a role in the pathology of CRC. [ABSTRACT FROM AUTHOR]

We develop a riboadenosine-substituted DNA probe-based bioluminescence method for real-time monitoring of exonuclease III (Exo III) activity and Exo III-assisted target recycling. This method employs the targetdependent ATP as the bioluminescence reporter without the involvement of any sophisticated luciferase manipulations, and the glow-type reaction kinetics facilitates the performance of imaging assay. [ABSTRACT FROM AUTHOR]

We develop a new method for the sensitive detection of polynucleotide kinase (PNK) using rolling circle amplification-induced chemiluminescence. This method exhibits high sensitivity with a detection limit of 2.20 x 10-4 U mL-1, which is superior to most reported approaches. Moreover, this method can be used to screen both the inhibitors and the activators of PNK, and can be further applied for real sample analysis. [ABSTRACT FROM AUTHOR]

We report for the first time the synthesis of CdSe@Fe2O3core/shell nanoparticles by a one-step seeded-growth approach. These nanoparticles not only retain their individual semiconducting and magnetic functionalties, but also exhibit some new properties which are affected by the coating components. These bifunctional CdSe@Fe2O3nanoparticles might find potential applications in biosensing and in biomedical research. [ABSTRACT FROM AUTHOR]

Mycotoxin contamination in food products may cause serious health hazards and economic losses. The effective control and accurate detection of mycotoxins have become a global concern. Even though a variety of methods have been developed for mycotoxin detection, most conventional methods suffer from complicated operation procedures, low sensitivity, high cost, and long assay time. Therefore, the development of simple and sensitive methods for mycotoxin assay is highly needed. The introduction of nucleic acid signal amplification technology (NASAT) into aptasensors significantly improves the sensitivity and facilitates the detection of mycotoxins. Herein, we give a comprehensive review of the recent advances in NASAT-based aptasensors for assaying mycotoxins and summarize the principles, features, and applications of NASAT-based aptasensors. Moreover, we highlight the challenges and prospects in the field, including the simultaneous detection of multiple mycotoxins and the development of portable devices for field detection. [ABSTRACT FROM AUTHOR]

The addition of Phase Change Materials (PCM) to the salt gradient solar pond can significantly increase the heat capacity of the solar pond. Paraffin wax has a suitable range of phase transitions temperature and is feasible to be applied in solar pond. However, the poor thermal conductivity of paraffin is detrimental to thermal storage and release. In this study, the heat transfer behavior for the composite PCM of paraffin and steel wires has been studied firstly, the results shows that adding steel wires in paraffin improve the heat transfer performance obviously. With 10wt% adding percent, the maximum of heat transfer coefficient is 18.17 W/m2 °C−1 with increment of 18.84 % in melting process; and the maximum is 21.59 W/m2 °C−1 with increment of 20.88% during freezing process. Secondly, experiment study has been conducted in an experimental solar pond, with 20 PCM capsule tubes added, adding RT 50 and RT 60 made the day–night temperature differences 2.87 °C and 2.53 °C smaller than that of without any PCM conditions. In this study, adding RT 50 and RT 60 in solar pond achieve 9.34% and 14.13% exergy increments respectively. The study suggested that adding PCM to solar pond improves the quality of the heat energy of solar pond, and further extends the heat application scope of solar pond.

As important post-transcriptional regulators, microRNAs (miRNAs) play irreplaceable roles in diverse cellular functions. Dysregulated miRNA expression is implicated in various diseases including cancers, and thus miRNAs have become the valuable biomarkers for disease monitoring. Recently, clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) system has shown great promise for the development of next-generation biosensors because of its precise localization capability, good fidelity, and high cleavage activity. Herein, we review recent advance in development of CRISPR/Cas-based biosensors for miRNA detection. We summarize the principles, features, and performance of these miRNA biosensors, and further highlight the remaining challenges and future directions. [ABSTRACT FROM AUTHOR]

Abstract Fuel cells have attracted increasing attention due to their low cost, high energy density, low environmental pollution, and abundant raw materials. Oxygen reduction reaction (ORR) is a core technology of fuel cells, and the development of new electrocatalysts with high ORR performance is highly desirable. Herein, we synthesize a series of B, N co‐doped hierarchical porous carbons using a soft template method with the integration of self‐assembly, calcination and etching. The obtained materials exhibit hierarchical porous structures, controllable pore distribution, partial graphite structures, and B, N co‐doping. They can function as the cost‐effective and metal‐free electrocatalysts, facilitating the diffusion of electrolyte ions and the improvement of ORR performance. Especially, the B, N co‐doped porous carbon with the B‐to‐N molar ratio of 5 (BNC‐5) displays a high ORR activity with a half‐wave potential (E1/2) of 0.73 V, an onset potential (Eonset) of 0.94 V, and a high limiting current density (JL) of 5.98 mA cm−2, superior to the N‐doped C (NC) and BNC‐1 (the B‐to‐N molar ratio=1), BNC‐3 (the B‐to‐N molar ratio=3) and BNC‐7 (the B‐to‐N molar ratio=7) under the identical conditions. Moreover, the BNC‐5 exhibits good cycling stability after 5000 cyclic voltammetry (CV) cycles and excellent tolerance toward even 3 M methanol. This research provides a new approach for the facile synthesis of dual element‐doped carbon electrocatalysts with high ORR performance.