Fluorescence imaging sheds light on the immune evasion mechanisms of hepatic stellate cells mediated by superoxide anion.

Whether and how the reactive oxygen species generated by hepatic stellate cells (HSCs) promote immune evasion of hepatocellular carcinoma (HCC) remains mysterious. Therefore, investigating the function of superoxide anion (O2•-), the firstly generated reactive oxygen species, during the immune evasion become necessary. In this work, we establish a novel in situ imaging method for visualization of O2•- changes in HSCs based on a new two-photon fluorescence probe TPH. TPH comprises recognition group for O2•- and HSCs targeting peptides. We observe that O2•- in HSCs gradually rose, impairing the infiltration of CD8+ T cells in HCC mice. Further studies reveal that the cyclin-dependent kinase 4 is deactivated by O2•-, and then cause the up-regulation of PD-L1. Our work provides molecular insights into HSC-mediated immune evasion of HCC, which may represent potential targets for HCC immunotherapy.

An efficient electrochemical biosensor for the detection of heavy metal lead in food based on magnetic separation strategy and Y-DNA structure.

Herein, an electrochemical biosensor was developed based on a magnetic separation strategy for the sensitive detection of the heavy metal Pb2+. The specific binding of Pb2+ and the aptamer (Apt) is used to trigger the release of the complementary chain (cDNA) on the magnetic bead system. The cDNA completes base complementary pairing with hairpins HP1 and HP2 at the electrode to form a Y-DNA structure. Then, the Y-DNA runs continuously with the assistance of the signal tag methylene blue (MB) and the current signal increases. However, in the absence of Pb2+, cDNA cannot be released and the Y-DNA structure cannot be formed on the electrode, resulting in a relatively low current signal. Under the optimal experimental conditions, the reduced peak current difference (ΔI) showed a good linear relationship with lg CPb2+ between 0.1 and 1000 nM, with a detection limit of 5.9 pM. In addition, the stability, reproducibility and detection capability of the sensors were investigated with satisfactory results.

A label-free impedance-based electrochemical sensor based on self-assembled dendritic DNA nanostructures for Pb2+ detection.

Here, a label-free impedance-based electrochemical sensor was developed for the quantitative detection of Pb2+. Using conductive gold nanomaterials as electrode substrate materials can provide sensors with larger specific surface area, action sites and excellent conductivity. DNA nanostructures are used for the determination of biomolecules due to their good properties. The Y-DNA structure is formed by the annealing of three DNA sequences, which acts as a stable structure and forms a dendritic structure in combination with the hybrid chain reaction. In the presence of the target Pb2+, it induces the conversion of specific aptamers into G-quadruplexes, resulting in HCR and Y-DNA loading on the electrodes and a significant change in the impedance value signal. Therefore, the proposed biosensor realizes the quantitative detection of Pb2+. Under the optimal experimental conditions, the concentration of Pb2+ exhibited a linear correlation range from 0.5 to1000 nmol/L with a limit of detection (LOD) of 0.38 nmol/L. The designed sensors have good recoveries in real samples (tap water and tea). This flexible experimental protocol has broad application prospects.