Identification, cleavage, and amplification (ICA): A versatile strategy for highly sensitive detection of miRNA.

MicroRNAs (miRNAs) are small RNA molecules that can play important roles as diagnostic/prognostic biomarkers and therapeutic targets for cancers and other diseases. Herein, an identification-cleavage-amplification (ICA) strategy for highly sensitive and versatile detection of miRNA has been proposed, and successfully applied to miR-155 and miR-21 assays. It combines an aligner-target mediated cleavage with strand displacement amplification (ATMC-SDA) to achieve the ICA process. During the identification process, a DNA-aligner (DA) and a DNA-amplicon (DM) can bind together with the help of target miRNA, forming a T-junction structure. Then, a nicking endonuclease (NEase), binding on the recognition sequence at the stem part of DA, can make a cleavage on DM, and the cleaved DM (CDM) can serve as an initiator to trigger the SDA reaction for signal amplification. Sharing the same set of enzymes and primers, the proposed ATMC-SDA can serve as a versatile ICA strategy for highly sensitive detection of various miRNAs, without the requirement of reverse transcription. Results show that the limits of detection (LOD) for miR-155 and miR-21 are 5.4 aM and 6.8 aM, respectively, with a dynamic range from 10.0 aM to 10.0 pM. The compatibility of ATMC-SDA with biological samples has also been tested by using human serum, indicating a promising potential for a wide variety of applications.

Cis-bifenthrin inhibits cortisol and aldosterone biosynthesis in human adrenocortical H295R cells via cAMP signaling cascade.

Cis-bifenthrin (cis-BF) is a common-used pyrethroid insecticide frequently detected in environmental and biological matrices. Mounting evidence highlights the endocrine disrupting effects of cis-BF due to anti-estrogenic or anti-androgenic activity. However, little is known about the exposure effects of cis-BF on adrenal cortex function. In this study, effects of cis-BF on biosynthesis of adrenal steroids, as well as the potential mechanisms were investigated in human adrenocortical carcinoma (H295R) cells. Cis-BF decreased basal production levels of cortisol and aldosterone, as well as cAMP-induced production of cortisol. Both he basal and cAMP-stimulated transcriptional levels of several steroidogenic genes were significantly down-regulated by cis-BF. As an important rate-limiting enzyme in steroidogenesis, the protein level of StAR was prohibited by cis-BF on both basal and cAMP-induced conditions. Intracellular level of cAMP was significantly reduced by cis-BF. Overall, these data suggest that cis-BF may inhibit the biosynthesis of cortisol and aldosterone via disrupting cAMP signaling cascade.

Structural properties, antioxidant and immune activities of low molecular weight peptides from soybean dregs (Okara).

In this study, a method for preparing low molecular weight peptides (HPH-VAP) from okara using high-pressure homogenization assisted double enzymes was proposed. In order to explore its advantages, the effects of various methods on protein extraction rate and on the structure, antioxidant and immune properties of peptides were compared. The results showed that the protein extraction rate of this method was increased by 69% and 51% compared with other methods, and the structure only led to changes in the hydrogen bonds between peptide chains. HPH-VAP was screened out through functional characteristics, its structure was identified by HPLC-MS/MS, and further immunological activity analysis was carried out. The results showed that it promoted cell phagocytic ability, NO level and release of cytokines IL-6, IFN- γ, TNF-α. Therefore, this method is an effective and applicable method for industrial preparation of okara peptides, and has a positive effect on the reuse of okara resources.

Gas-solid phase flow synthesis of Cu-Co-1,3,5-benzenetricarboxylate for electrocatalytic oxygen evolution.

Using an environmentally friendly method to produce a stable and highly catalytically active electrocatalyst for the oxygen evolution reaction (OER) is becoming increasingly urgent. Herein, a novel bimetallic metal-organic framework (MOF), specifically a copper-cobalt 1, 3, 5-benzenetricarboxylate (Cu-Co-BTC) MOF, was successfully prepared by employing the gas-solid two-phase flow (GSF) synthetic technique. The as-prepared Cu-Co-BTC with its multiple active sites afforded a current density of 10 mA cm-2 at 239 mV for the OER in a 1 mol L-1 KOH solution, and showed a better electrocatalytic performance than did single-metal-containing Cu-BTC and Co-BTC materials. This work provides a new idea, one involving using novel gas-solid phase reactions for the preparation of electrocatalysts in large quantities.

Soft nanomaterial-based targeting polymersomes for near-infrared fluorescence multispectral in vivo imaging.

We report here the soft nanomaterial-based targeting polymersomes for near-infrared (NIR) fluorescence imaging to carry out in vivo tumor detection. Two polymersome-based NIR fluorescent probes were prepared through the self-assembly of amphiphilic block copolymers, poly(butadiene-b-ethylene oxide) (PEO-b-PBD). Each of them was encapsulated with distinct hydrophobic near-infrared dyes (DiD and DiR) and modified with different targeting ligands (anti-CEA antibody and anti-EGFR antibody), respectively. After simultaneous injection of these two probes into the tumor-bearing mice via tail vein, multispectral near-infrared fluorescence images were obtained. The results indicate that both probes are successfully directed to the tumor foci, where two distinguishable fluorescent signals were detected through the unmixed fluorescence images. By taking advantage of two targeting polymersome-based probes with distinct fluorescent features, the proposed multispectral near-infrared fluorescence imaging method can greatly improve the specificity and accuracy for in vivo tumor detection.

Peptide-induced fluorescence quenching of conjugated polyelectrolyte for label-free, ultrasensitive and selective assay of protease activity.

We report here a label-free method for ultrasensitive and selective assay of protease activity based on the peptide-induced fluorescence quenching of conjugated polyelectrolyte (PPESO(3)). It is very interesting to find that there is a critical length of oligo-polyarginine (i.e., Arg(5)) below which 1) the quenching efficiency of PPESO(3) is sharply decreased, and more importantly, 2) the trypsin-catalyzed hydrolysis is greatly slowed down. This opens good opportunities for not only the ultrasensitive assay of trypsin, but the specific detection of other proteases if carefully designing an appropriate peptide length and the cleavage site. Herein, the enzyme selected as a proof of concept is chymotrypsin. Due to the essence that any cleavage of the designed peptide probes will result in a notable decrease or even a complete loss of their capability to quench the emission of PPESO(3), the limits of detection for trypsin and chymotrypsin have been found as low as 0.25 ng/mL (11 pM) and 0.15 ng/mL (6 pM), respectively. Both are superior to those of most previous methods by 1-2 orders or higher.