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DNA (cytosine-5)-methyltransferase1 (DNMT1) is the most abundant DNA methyltransferase in somatic cells, and it plays an important role in the initiation, occurrence, and rehabilitation of tumors. Herein, we developed a novel strategy for the detection of the level of DNMT1 in human plasma using the self-assembled nucleic acid probe signal amplification technology. In this method, the DNMT1 monoclonal antibody (McAbDNMT1) was immobilized on carboxyl magnetic beads to form immunomagnetic beads and then captured DNMT1 specifically. After that, DNMT1 polyclonal antibody (PcAbDNMT1) and biotinylated sheep anti-rabbit IgG (sheep anti rabbit IgG-Biotin) were sequentially added into the system to react with DNMT1 and form biotinylated double antibody sandwich immunomagnetic beads. In the presence of the bridging medium streptavidin, the biotinylated double antibody sandwich immunomagnetic beads would form a complex with biotinylated poly-fluorescein (Biotin-poly FAM), and the fluorescence intensity of the complex was proportional to the concentration of DNMT1. Immunomagnetic beads can capture the target DNMT1 in the sample, and Biotin-poly FAM can realize signal amplification. Using these strategies, we got a linear range of the system for DNMT1 level detection was from 2 nmol/L to 200 nmol/L, and the limit of detection (LOD) was 0.05 nmol/L. The method was successfully applied for the determination of DNMT1 in human plasma with the recovery of 101.3-106.0%. Therefore, this method has the potential for the detection of DNMT1 level in clinical diagnosis.Hypochlorite (ClO-), a type of reactive oxygen species (ROS), plays an essential role in complex biological systems. Real-time detection of the content and distribution of ClO- in cells or subcellular organelle is critically essential. In this paper, a lysosomal-targeted fluorescent probe, Cou-Lyso, was constructed for real-time detection of ClO- in a ratiometric manner, achieving high sensitivity with a low detection limit (0.58 μM). Upon reaction with ClO-, this probe was subjected to a significant fluorescence change from red emission (λmaxem = 610 nm) to green emission (λmaxem = 535 nm) with the ratio of I535 nm/I610 nm displaying a 76-fold enhancement from 0.04 to 3.03. The confocal imaging experiments for Cou-Lyso showed that this probe could detect ClO- in living cell and zebra fish. This probe has been successfully applied to stain lysosome and image lysosomal ClO- based on co-localization imaging experiments.A novel visual nanoprobe was developed for the sequential detection of morin and zinc ion (Zn2+) based on Cl and N co-doped carbon quantum dots (ClNCQDs) via a fluorometric and colorimetric dual-readout mode. The yellow fluorescence ClNCQDs was synthesized by the one-step hydrothermal treatment of o-chlorobenzoic acid and p-phenylenediamine. The most distinctive property of the ClNCQDs is the large stokes shift (177 nm), which is significantly higher than other reported CQDs. The fluorescence of the ClNCQDs can be effectively quenched by morin based on the synergistic effect of IFE, electrostatic interaction, and dynamic quenching process, and recovered upon the addition of Zn2+ due to strong interaction between morin and Zn2+. The nanoprobe exhibited favorable selectivity and sensitivity toward morin and Zn2+ with detection limits of 0.09 µM and 0.17 µM, respectively. Simultaneously, the color of the ClNCQDs solution was changed (light-pink → faint-yellow → dark-yellow) along with the variation of the fluorescence signal of the ClNCQDs. This proposed nanoprobe was successfully applied for morin and Zn2+ analyses in actual samples and live cells with high accuracy. The results of this study demonstrate the great application prospects of the ClNCQDs for morin and Zn2+ detection in complex actual samples and biosystems.A new pH-sensitive fluorescent probe NAP-MDA was designed and synthesized. NAP-MDA consists of 1,8-naphthalimide as fluorophore, morpholine and N,N-dimethylethylenediamine as pH-responsive groups. Due to the photoinduced electron transfer (PET) mechanism, the fluorescence of 1, 8-naphthalimide was thoroughly quenched under alkaline condition (pH > 10.0), however, NAP-MDA displayed increasing fluorescence as the rise of acidity. Notably, NAP-MDA possessed an excellent linear dependence with neutral to alkaline pH (7.2-9.4), with a pKa of 8.38. NAP-MDA had good photostability and reversibility. Meanwhile, the probe was selective to pH without interference from common reactive species, temperature and viscosity. Fluorescent testing strips were fabricated with NAP-MDA and were successfully utilized to visualize the different pH with a handhold UV lamp. Confocal fluorescence imaging in live cells demonstrated that NAP-MDA mainly fluoresced in lysosomes, and could be applied for quantification of the pH within live cells.

Exosomes in the tumor microenvironment (TME) facilitate tumor progression by enabling inter-cellular communication. Tumor cell-derived exosomes can polarize tumor-associated macrophages (TAMs) to an immunosuppressive M2 phenotype. The aim of this study was to determine the role of exosomal circFARSA in non-small cell lung cancer (NSCLC) and elucidate the underlying mechanisms.

In situ circFARSA expression in NSCLC tissues was analyzed using qRT-PCR. The in vitro migration of NSCLC cells was evaluated using a transwell assay or through indirect co-culture with M2 macrophages, as appropriate. Immunoprecipitation (IP), western blotting, RNA binding protein immunoprecipitation (RIP), and RNA pull down assays were conducted for mechanistic studies.

CircFARSA was significantly upregulated in NSCLC tissues, and the ectopic overexpression of circFARSA enhanced NSCLC cell metastasis. Furthermore, NSCLC cell-derived exosomal circFARSA polarized the macrophages to a M2 phenotype. The NSCLC cells co-cultured with macrophages transfected with circFARSA or pre-treated with exosomal circFARSA showed enhanced EMT and metastasis. Mechanistically, exosomal circFARSA induced M2 polarization via PTEN ubiquitination and degradation, which further activated the PI3K/AKT signaling pathway. see more In addition, eIF4A3 promoted circRNA biogenesis and cyclization by binding to its flanking sequences.

Exosomal circFARSA plays a crucial role in cross-talk between macrophages and NSCLC cells through the PTEN/PI3K/AKT signaling pathway, and is a promising diagnostic/prognostic biomarker for NSCLC.

Exosomal circFARSA plays a crucial role in cross-talk between macrophages and NSCLC cells through the PTEN/PI3K/AKT signaling pathway, and is a promising diagnostic/prognostic biomarker for NSCLC.