Multiplex detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

A simple, highly sensitive, and specific assay was developed for the homogeneous and multiplex detection of microRNAs (miRNAs) by combining molecular beacon (MB) probes and T7 exonuclease-assisted cyclic amplification. An MB probe with five base pairs in the stem region without special modification can effectively prevent the digestion by T7 exonuclease. Only in the presence of target miRNA is the MB probe hybridized with the target miRNA, and then digested by T7 exonuclease in the 5' to 3' direction. At the same time, the target miRNA is released and subsequently initiates the nuclease-assisted cyclic digestion process, generating enhanced fluorescence signal significantly. The results show that the combination of T7 exonuclease-assisted cyclic amplification reaction and MB probe possesses higher sensitivity for miRNA detection. Moreover, multiplex detection of miRNAs was successfully achieved by designing two MB probes labeled with FAM and Cy3, respectively. As a result, the method opens a new pathway for the sensitive and multiplex detection of miRNAs as well as clinical diagnosis. Graphical Abstract A simple, highly sensitive, and specific assay was developed for the detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

Light-Excited Antibiotics for Potentiating Bacterial Killing via Reactive Oxygen Species Generation.

The irrational or excessive use of antibiotics causes the emergence of bacterial resistance, making antibiotics less effective or ineffective. As the number of resistant antibiotics increases, it is crucial to develop new strategies and innovative approaches to potentiate the efficacy of existing antibiotics. In this paper, we report that some existing antibiotics can produce reactive oxygen species (ROS) directly under light irradiation. Thus, a novel antibacterial photodynamic therapy (PDT) strategy is proposed by using existing antibiotics for which the activities are potentiated via light-activation. This antibiotic-based PDT strategy can achieve efficient bacteria killing with a low dosage of antibiotics, indicating that bacterial killing can be enhanced by the light-irradiated antibiotics. Moreover, the specific types of ROS produced by different antibiotics under light irradiation were studied for better elucidation of the antibacterial mechanism. The findings can extend the application of existing antibiotics and provide a promising strategy for treatment of bacterial infections and even cancers.

Conjugated Polymers Act Synergistically with Antibiotics to Combat Bacterial Drug Resistance.

The emergence of drug-resistant bacteria severely challenges the antimicrobial agents and antibacterial strategy. Here, we demonstrate a novel, simple, and highly efficient combination therapy strategy by direct combinations of cationic conjugated polymers (CCPs) with polypeptide antibiotics against Gram-negative and Gram-positive bacteria based on a synergistic antibacterial effect. The combination therapy method enhances the antibacterial efficacy with a significantly reduced antibiotic dosage. Also, the highly efficient and synergistic killing of drug-resistant bacteria is realized. Using combinations of CCPs and antibiotics to show increased antibacterial activity, this strategy will provide a much wider scope of the discovery of efficient antibacterial systems than that of antibiotic-antibiotic combinations. The proposed combination therapy method provides a universal and powerful platform for the treatment of pathogens, in particular, the drug-resistant bacteria, and also opens a new way for the development of efficient antibacterial systems.