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Objective: To establish a method for Point of Care Testing detection Candida mannan IgG antibody based on chemiluminescence immunoassay. Methods: The candida mannan IgG antibody detection system was established according to the indirect method principle, and the analytical performance was evaluated according to the approved guidelines of the American Clinical Laboratory Standardization Association (CLSI). Results: Biotinylated Candida mannan antigen and Alkaline Phosphatase-labeled IgG antibody was successfully conjugated, and the optimal reaction concentration was 1 μg/ml. The limit of blank was 5.20 AU/ml. The limit of detection was 5.50 AU/ml. The precision and repeatability of CV were less than 10%. There was no cross reaction between the detection method and common specific antibodies. Hemoglobin, triglyceride and bilirubin had no significant effect on clinical samples. The overall coincidence rate between the detection method and ELISA were 90%, and the Kappa value=0.80, regression analysis and deviation analysis showed that the two methods had strong consistency (R²=0.989 8). Conclusion: The performance of the strip chemiluminescence detection system established in this study meets the relevant requirements of clinical auxiliary diagnostic technology, and is suitable for the rapid detection of Candida mannan IgG antibody in serum samples. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects and mechanisms of exogenous calcium-binding protein S100A4 on proliferation, survival and migration functions of glioma cells. Methods: Pan-cancer dataset was downloaded from UCSC database for the analysis of S100A4 expression and prognosis in pan-cancer, and transcriptome and clinical data of glioma patients were downloaded from CGGA database. Prognosis and progression of S100A4 expression in glioma patients were analyzed by R software. S100A4 protein network interaction of glioma patients were addressed by online analytical tools GEPIA and STRING. Human glioma cell lines (U87 and U251) were cultured in vitro, and the experiment was divided into 3 groups: PBS group, S100A4 group and S100A4+TAK242 group [Resatorvid (TAK242) was a specific inhibitor of Toll-like receptors 4 (TLR4)]. Flow cytometry was used to detect proliferation and apoptosis of glioma cells. Wound healing assay, Transwell assay and clone formation assay were used to detect migration and proliferation ability of U87 and U251 cells, and Western blot was used to detect levels of TLR4 protein and NF-κB related signaling proteins. Results: Bioinformatics results showed that S100A4 was significantly upregulated in multiple tumours (P<0.05), which included glioblastoma (GBM), lower grade glioma (LGG), stomach adenocarcinoma (STAD), liver hepatocellular carcinoma (LIHC), etc, with poorer prognosis in GBM and LGG. Compared with glioma patients with low expression of S1004, high expression S100A4 in glioma patients had shorter survival and higher degree of WHO classification. In addition, S100A4 protein in glioma patients may interact with Annexin A2 (ANXA2), TLR4 and advanced glycosylation end product-specific receptor (AGER) proteins. In cellular experiments, U87 and U251 cells showed enhanced proliferation and migration, and significantly up-regulated levels of TLR4, p-ERK1/2, p-p38 and p-p65 proteins after exogenous S100A4 treatment compared to PBS group (P<0.05), while glioma cells in S100A4+TAK242 group showed weaker proliferation and migration, and lower TLR4, p-p38 and p-p65 protein levels than those in S100A4 group, TLR4, p-ERK1/2, p-p38, p-p65 protein levels were significantly down-regulated (P<0.05). Conclusion: S100A4 may regulate function of glioma proliferation, migration and survival through TLR4/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of dexmedetomidine (DEX) on the malignant biological behavior of gastric cancer (GC) cells through the immune regulation mechanism mediated by cyclic GMP-AMP synthase-stimulator of interferon gene (cGAS-STING) pathway. Methods: GC cell line MGC-803 was randomly divided into Control group (blank medium treatment), DEX low concentration group (DEX-L group, 1 ng/ml), DEX medium concentration group (DEX-M group, 10 ng/ml), DEX high concentration group (DEX-H group, 100 ng/ml) and DEX high concentration+cGAS inhibitor RU.521 group (DEX-H+RU.521 group, 100 ng/ml DEX+1.0 μmol/L RU.521). Cell proliferation was detected by CCK-8 method. Cell scratch test was used to detect the migration ability of cells in each group. Transwell test was used to detect the invasive ability of cells in each group. The apoptosis rate was detected by flow cytometry. The levels of IL-2, interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in cells were detected by ELISA. Real time-fluorescent quantitative PCR (RT-qPCR) was applied to detect the expression levels of cGAS, STING and interferon type I (IFN-I) mRNA. Western blot was used to detect the expressions of cGAS, STING, Bax, CyclinD1, matrix metalloproteinase 9 (MMP9), N-cadherin, Vimentin, E-cadherin, Caspase3, Caspase8 and their shear type and phosphorylation level of TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3). Results: Compared with Control group, the cell migration rate, number of cell invasions, TNF-α level, CyclinD1, MMP9, N-cadherin, Vimentin protein expressions in MGC-803 cells in DEX-M and DEX-H groups were decreased obviously (P<0.05), the growth inhibition rate (48 h, 72 h), apoptosis rate, IL-2, IFN-γ, Bax, E-cadherin, Cleaved Caspase3, Cleaved Caspase8 protein expression levels, cGAS, STING, IFN-I mRNA levels and protein expression levels and phosphorylation levels of TBK1 and IRF3 were increased obviously (P<0.05). RU.521 weakened the inhibitory effects of DEX on the proliferation, migration and invasion of GC cells and the ability to induce apoptosis, and alleviated the improvement on immune function. Conclusion: DEX may inhibit the proliferation, migration and invasion of GC cells and induce apoptosis of GC cells by activating cGAS-STING pathway mediated immune regulation. [ABSTRACT FROM AUTHOR]