Prognostic Value and Immunological Role of P4HA3 in Colon Adenocarcinoma.

Purpose: Prolyl 4-hydroxylase subunit alpha 3 (P4HA3) has been proven to participate in the occurrence and development of multiple cancers. However, the functional role of P4HA3 in the tumor immune microenvironment (TIME) of colon adenocarcinoma (COAD) and the prognosis of COAD patients has not been clarified. This study aimed to elucidate the immunological role and prognostic value of P4HA3 in COAD. Methods: P4HA3 expression in COAD tissues was analyzed via experiments and a bioinformatics algorithm. Based on the COAD patients in The Cancer Genome Atlas database, we comprehensively evaluated whether the expression levels of P4HA3 affected clinical prognosis, TIME, and immunotherapy of COAD using the R platforms and several public databases, including GEPIA, TIMER, TISIDB, and TCIA. Results: The results of the pan-cancer analysis indicated that P4HA3 expression was significantly different in most tumor tissues compared with normal tissues. P4HA3 was overexpressed in COAD tissues, and overexpression of P4HA3 was associated with a worse overall survival and a shorted progression-free interval in COAD patients. The expression of P4HA3 was positively correlated with pathological stage, T stage, N stage, perineural infiltration, and lymphatic infiltration. There were significant correlations of P4HA3 expression levels with immune cell infiltration and their makers, as well as immunomodulators, chemokines, and microsatellite status. Moreover, overexpression of P4HA3 was associated with a lower response rate to immunotherapy in the IMvigor210 cohort. Conclusion: Overexpression of P4HA3 is closely related to the poor prognosis of COAD patients, and P4HA3 is a potential target for immunotherapy in COAD patients.

Effects of Resveratrol on Tight Junction Proteins and the Notch1 Pathway in an HT-29 Cell Model of Inflammation Induced by Lipopolysaccharide.

Ulcerative colitis (UC) is closely associated with disruption of intestinal epithelial tight junction proteins. A variety of studies have confirmed that resveratrol (RSV), a natural polyphenolic compound, has a potential anti-inflammatory effect and can regulate the expression of tight junction proteins. However, the mechanism by which RSV regulates the expression of tight junction proteins in the intestinal epithelium remains unclear. Therefore, we investigated the potential effect of RSV on tight junction proteins in an HT-29 cell model of inflammation induced by lipopolysaccharide (LPS) and explored its mechanism of action. First, the downregulated expression of the tight junction proteins occludin, ZO-1, and claudin-1 in the HT-29 cell model of inflammation induced by LPS was reversed by incubation with RSV, accompanied by a decrease in the expression of tumor necrosis factor α-converting enzyme (TACE). Additionally, the Notch1 pathway was attenuated and the expression of the inflammatory factors IL-6 and TNF-α was decreased by treatment with RSV. Second, after Jagged-1 was used in combination with RSV to reactivate the Notch1 pathway, the protective effects of RSV against the LPS-induced reductions in the expression of the tight junction proteins occludin, ZO-1, and claudin-1 and the decreases in the levels of the inflammatory factors IL-6 and TNF-α were abolished. These results suggest that RSV might regulate the expression of tight junction proteins by attenuating the Notch1 pathway.

Optimization of GPC3-specific chimeric antigen receptor structure and its effect on killing hepatocellular carcinoma cells.

To investigate the effect of optimized GPC3-specific chimeric antigen receptor (GPC3-CAR) structure on killing hepatocellular carcinoma (HCC) cells. We constructed three lentiviral expression vectors with different CAR structures by genetic engineering and molecular cloning techniques. These three CAR structures shared the same intracellular signaling region consisting of 4-1BB and CD3ζ, but had different hinge and transmembrane regions. Specifically, GPC3-O4-CAR contained an optimized CD8α hinge region and a 4-1BB transmembrane domain; GPC3-CD8-CAR contained an optimized CD8α hinge region and a CD8α transmembrane domain; and GPC3-ori-CAR contained an original CD8α hinge region and a 4-1BB transmembrane domain. With similar transfection efficiency, it was observed by fluorescence microscopy that GPC3-O4-CAR expression on the surface of 293 T cells was much higher than those of the other two. Cytotoxicity experiments showed that T or NK cells with GPC3-O4-CAR structure were more lethal and could secrete more IFN-γ than the other two. In conclusion, GPC3-O4-CAR can be efficiently and stably expressed on the cell surface. Moreover, both the killing effect of transduced T and NK cells on GPC3-positive HCC cells and release of IFN-γ are increased.

Quantitative analysis of source and fate of N-nitrosamines and their precursors in an urban water system in East China.

The source and fate of N-nitrosamines and their precursors in terms of formation potential (FP) was investigated quantitatively in the city level for the first time. Different sources of nitrosamines and their precursors were investigated in one city in the Yangtze River delta, China. The source water located downstream of the city contained 8.4 ng/L of N-nitrosodimethyamine (NDMA) and 153 ng/L of NDMA FP. The contribution of each discharge source was evaluated based on the concentration, the river water flux, and the amount of wastewater discharges. Textile printing and dyeing wastewater, and electroplating industrial wastewater contained high concentration of nitrosamines and were important discharge sources. Taking NDMA and NDMA FP attenuation by photolysis and biodegradation into consideration, the mass load calculation showed upstream surface water brought about 13 ± 4% of NDMA and 21 ± 3% of NDMA FP to downstream source water. Local wastewater discharges contributed 30 ± 8% of NDMA and 17 ± 2% of NDMA FP to downstream source water. Endogenous formation via amino acids metabolism could contribute 36% of NDMA FP (maximum) to downstream source water. Overall, this study provides a protocol for quantitative evaluation of the nitrosamine contribution to urban water supply from different contamination sources.