Search

Poor vacuum stability limits the application of many matrices in matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) that requires long-term measurement duration in high vacuum. In this study, a new approach using conjugate polymer anchor to protect unstable matrix from volatilizing in MALDI source based on ion bond is provided. Unlike strong covalent bonds which often introduce unnecessary groups, the weaker ion bonds are more conducive to breaking under laser radiation while effectively preventing matrix volatilization in a vacuum environment. The results confirm that conjugate polymer anchor will neither introduce additional ion peaks nor affect signal intensity, yet maintains comparable quantification properties. Vacuum stability of three kinds of typical matrices is enhanced using polymer anchors, and the in situ MALDI MS imaging of mouse brain and liver cancer is improved significantly., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Introduction: Hepatocellular carcinoma (HCC) has very poor prognosis due to its immunosuppressive properties. An effective measure to regulate tumor immunity is brachytherapy, which uses 125 I seeds planted into tumor. T cell immune receptors with immunoglobulin and ITIM domains (TIGIT) is highly expressed in HCC. The TIGIT-targeted probe is expected to be an effective tool for indicating immunomodulation of 125 I seed brachytherapy in HCC. In this study, We constructed a novel peptide targeting TIGIT to evaluate the immune regulation of 125 I seed brachytherapy for HCC by near-infrared fluorescence (NIRF)., Methods: Expression of TIGIT by immunofluorescence (IF) and flow cytometry (FCM) in different part and different differentiated human liver cancer tissues was verified. An optical fluorescence probe (Po-12) containing a NIRF dye and TIGIT peptide was synthesized for evaluating the modulatory effect of 125 I seed brachytherapy. Lymphocytes uptake by Po-12 were detected by FCM and confocal microscopy. The distribution and accumulation of Po-12 in vivo were explored by NIRF imaging in subcutaneous and orthotopic tumors. IHC and IF staining were used to verify the expression of TIGIT in the tumors., Results: TIGIT was highly expressed in HCC and increased with tumor differentiation. The dye-labeled peptide (Po-12) retained a stable binding affinity for the TIGIT protein in vitro . Accumulation of fluorescence intensity (FI) increased with time extended in subcutaneous H22 tumors, and the optimal point is 1 h. TIGIT was highly expressed on lymphocytes infiltrated in tumors and could be suppressed by 125 I seed brachytherapy. Accumulation of Po-12-Cy5 was increased in tumor-bearing groups while declined in 125I radiation group., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zeng, Shen, Shu, Min, Shu, Yao, Wang and Chen.)

Articular cartilage injury and therapy are important clinical issues around the world. Mesenchymal stem cells (MSCs) have the ability to differentiate into chondrocytes, which makes MSCs good candidates for use in cartilage repairing. However the regulation and the mechanism of chondrogenesisin MSCs is still unclear. To clarify the factor and mechanism which contribute to the process of chondrogenic differentiation, we focus on miRNAs. Considering the role of miR-181a in chondrogenesis and osteoblast formation, we tested the expression of miR-181a in the induced chondrogenic differential pig PBMSCs by using qRT-PCR. And we identified miR-181a as an up-regulated miRNA in the TGF-β3-induced pig PBMSCs chondrogenic differentiation from the early stages and maintained elevated throughout the whole process. After inhibition of the endogenesis miR-181a expression by transfecting the miR-181a inhibitor, the western-blot results and immunofluorescence results indicated that the expression of differentiation-related protein COL2A1, BMP2 were decreased, together with the Alcian blue assay, proving the process of differentiation was inhibited significantly. Taken together, our results demonstrated that miR-181a might be necessary in chondrogenesis of MSCs. Even so, the mechanism of miR-181a on regulating the chondrogenesis still needed to be investigated in future work. And our data would provide an experimental evidence for the research of tissue engineering., Competing Interests: None., (IJCEP Copyright © 2018.)