A rapid and sensitive fluorescent chromatography with cloud system for MPXV point-of-care diagnosis.

Monkeypox (mpox) is spreading around the world, and its rapid diagnosis is of great significance. In the present study, a rapid and sensitive fluorescent chromatography assisted with cloud system was developed for point-of-care diagnosis of mpox. To screen high affinity antibodies, nanoparticle antigen AaLS-A29 was generated by conjugating A29 onto scaffold AaLS. Immunization with AaLS-A29 induced significantly higher antibody titers and monoclonal antibodies were generated with the immunized mice. A pair of monoclonal antibodies, MXV 14 and MXV 15, were selected for fluorescence chromatography development. The Time-Resolved Fluorescence Immunoassay (TRFIA) was used to develop the chromatography assay. After optimization of the label and concentration of antibodies, a sensitive TRFIA assay with detection limit of 20 pg/mL and good repeatability was developed. The detection of the surrogate Vaccinia virus (VACA) strain Tian Tan showed that the TRFIA assay was more sensitive than the SYBR green I based quantitative PCR. In real samples, the detection result of this assay were highly consistent with the judgement of Quantitative Real-Time PCR (Concordance Rate = 90.48%) as well as the clinical diagnosis (Kappa Value = 0.844, P < 0.001). By combining the portable detection and online cloud system, the detection results could be uploaded and shared, making this detection system an ideal system for point-of-care diagnosis of mpox both in field laboratory and outbreak investigation.

Tannic Acid Accelerates Cutaneous Wound Healing in Rats Via Activation of the ERK 1/2 Signaling Pathways.

Objective: This study was aimed to evaluate the effect of tannic acid (TA), a natural plant polyphenol astringent, on wound healing in vitro and in vivo, and to elucidate the underlying molecular signaling pathway in the wound healing. Approach: Cutaneous skin wounds were created in rats and then treated until closure with purified TA, serum or tissue samples were collected to test the concentration of factors by enzyme-linked immunosorbent assay (ELISA), and the expression in gene or protein was measured by quantitative real-time polymerase chain reaction or Western blot. We explored the cell-/dose-specific responses of TA (0.1-0.4 µg/mL) on proliferation and gene and protein expression of fibroblast NIH 3T3 cells. Results: The wounds on rats treated by TA got healed faster than those in the untreated group. The histopathology study showed that TA accelerated re-epithelialization and increase in hair follicles could be detected. The levels of growth factors including basic fibroblast growth factor (bFGF), transforming growth factor-beta, and vascular endothelial growth factor in TA-treated groups were all increased, and the content of interleukin-1 (IL-1) and IL-6 was decreased significantly when compared with that of the untreated group. The NIH 3T3 cells grow faster in 6 h at concentration of 0.1 µg/mL, and the expression of bFGF in gene and protein was increased significantly in the 0.1 µg/mL TA group. Further study revealed that the protein levels of bFGF, extracellular signal regulated kinase (Erk) 1/2, and P-Erk 1/2 in Erk 1/2 pathway were increased after TA treatment. Innovation: The role of TA in wound healing efficacy is unclear; this study, therefore, assesses the effects of TA on wound healing in different periods and the underlying molecular mechanisms. Conclusion: These results suggested that TA could accelerate wound healing through modulation of inflammatory cytokines and growth factors and activate Erk 1/2 pathway. In conclusion, TA may be a potential agent in promoting wound healing.