Ratiometric fluorescence and visual determination of tetracycline antibiotics based on Y3+ and copper nanoclusters-induced cascade signal amplification.

A ratiometric fluorescence probe is proposed for sensitive and visual detection of tetracyclinee (TC) based on cascade fluorescence signal amplification induced by bovine serum albumin-stabilized copper nanoclusters (BSA-CuNCs) and yttrium ions (Y3+). TC can combine with Y3+ to form the complex (TC-Y3+) to enhance the fluorescence of TC at 515 nm. Then, positively charged TC-Y3+ and negatively charged BSA-CuNCs was bonded together by electrostatic interactions to achieve the fluorescence resonance energy transfer (FRET) process. With the increase of TC concentration, the fluorescence intensity of TC-Y3+ at 515 nm (F515) gradually increased; meanwhile, the fluorescence intensity of BSA-CuNCs at 405 nm (F405) decreased gradually. The ratio of F515 and F405 was used for the quantitative determination of TC. The linear range of the constructed fluorescent probe is 1.0 to 60.0 µM, and the limit of detection is 0.22 µM. The method was successfully applied to the determination of TC in spiked milk with recoveries ranging from 94.3 to 112%. Furthermore, the color of this platform can be observed from dark violet to bright green under the UV lamp. Since the response time of the reaction is less than 10 s, an intelligent sensing platform based on the use of the smartphone as image acquisition equipment was also established to realize rapid on-site and portable detection of TC through the colorimetric recognition application.

Detection of fucosylated extracellular vesicles miR-4732-5p related to diagnosis of early lung adenocarcinoma by the electrochemical biosensor.

Our preliminary investigation has identified the potential of serum fucosylated extracellular vesicles (EVs) miR-4732-5p in the early diagnosis of lung adenocarcinoma (LUAD) by a fucose-captured strategy utilizing lentil lectin (LCA)-magnetic beads and subsequent screening of high throughput sequencing and validation of real-time quantitative polymerase chain reaction (RT-qPCR). Considering the relatively complicated procedure, expensive equipment, and stringent laboratory condition, we have constructed an electrochemical biosensor assay for the detection of miR-4732-5p. miR-4732-5p is extremely low in serum, down to the fM level, so it needs to be detected by highly sensitive electrochemical methods based on the Mg2+-dependent DNAzyme splitting nucleic acid lock (NAL) cycle and hybridization chain reaction (HCR) signal amplification. In this study, signal amplification is achieved through the dual amplification reactions using NAL cycle in combination with HCR. In addition, hybridized DNA strands bind to a large number of methylene blue (MB) molecules to enhance signaling. Based on the above strategy, we further enhance our signal amplification strategies to improve detection sensitivity and accuracy. The implementation of this assay proceeded as follows: initially, miR-4732-5p was combined with NAL, and then Mg2+-dependent DNAzyme splitted NAL to release auxiliary DNA (S1) strands, which were subsequently captured by the immobilized capture probe DNA (C1) strands on the electrode surface. Following this, abundant quantities of DNA1 (H1) and DNA2 (H2) tandems were generated by HCR, and S1 strands then hybridized with the H1 and H2 tandems through base complementary pairing. Finally, MB was bonded to the H1 and H2 tandems through π-π stacking interaction, leading to the generation of a signal current upon the detection of a potential capable of inducing a redox change of MB by the electrode. Furthermore, we evaluated the performance of our developed electrochemical biosensor assay. The results demonstrated that our assay is a reliable approach, characterized by its high sensitivity (with a detection limit of 2.6 × 10-17 M), excellent specificity, good accuracy, reproducibility, and stability. Additionally, it is cost-effective, requires simple operation, and is portable, making it suitable for the detection of serum fucosylated extracellular vesicles miR-4732-5p. Ultimately, this development has the potential to enhance the diagnostic efficiency for patients with early-stage LUAD.

Antibacterial mechanism of polysaccharides from the leaves of Lindera aggregata (Sims) Kosterm. by metabolomics based on HPLC/MS.

Lindera aggregata (Sims) Kosterm. is a traditional Chinese herb, which has been proven to have excellent antibacterial activity. In this work, we firstly extracted the polysaccharides from the leaves of Lindera aggregata (Sims) Kosterm. (LLPs), and explored their antibacterial activity and related mechanisms. The experimental results show that LLPs are a good antibacterial agent, which can damage the cell structure of bacteria and lead to the leakage of intracellular lysates. Compared with Escherichia coli (E. coli), LLPs showed better inhibitory activity against Staphylococcus aureus (S. aureus). Furthermore, the administration of LLPs not only led to the upregulation of the levels of fructose-1,6-bisphosphate (F-1,6-P) and citric acid in the glycolysis and tricarboxylic acid cycle pathways in bacteria, but also resulted in the down-regulation of the levels of oxaloacetate (OAA) and 1,3-diphosphoglycerate (1,3-BPG). This study confirmed that LLPs have good antibacterial activity, and broaden the application of the leaves of Lindera aggregata (Sims) Kosterm. in the antibacterial field. It provides ideas for exploring the antibacterial mechanism of active ingredients of Chinese herbal medicine through metabolomics.