RIPK4 Is an Immune Regulating-Associated Biomarker for Ovarian Cancer and Possesses Generalization Value in Pan-Cancer.

Ovarian cancer (OC) is the most lethal gynecologic cancer. Many studies have reported that RIPK4 (receptor interacting serine/threonine kinase 4) displayed a dysregulated level in many types of tumors. However, its expressions and functions in OC were rarely reported. The levels of RIPK4 were detected in OC and nontumor specimens using TCGA and GEO datasets. The prognostic values of RIPK4 in patients were determined using Kaplan-Meier methods and Kaplan-Meier assays. GO assays and KEGG pathway assays were carried out for functional enrichments. CIBERSORT was applied for estimating the fractions of immune cell types. Finally, RIPK4 was validated in pan-cancer. In this study, our group found that RIPK4 exhibited a higher level of RIPK4 in OC specimens than nontumor specimens. Survival studies revealed that patients with high RIPK4 expressions showed a shorter overall survival than those with low RIPK4 expression. Multivariate assays further confirmed that RIPK4 expression was an independent prognostic element for OC. KEGG pathway analysis displayed that the dysregulated genes in specimens with high RIPK4 expressions were enriched in focal adhesion, proteoglycans in cancer, central carbon metabolism in cancer, and insulin secretion. Correlation analyses showed that several TICs were positively correlated with RIPK4 expression. The pan-cancer validation results showed that RIPK4 was associated with survival in five tumors. Overall, our findings suggested RIPK4 as a prognostic marker in OC.

Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification.

OBJECTIVES: To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification. METHODS: The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained. RESULTS: There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/µL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/µL, and the discrimination accuracy was 100%. CONCLUSIONS: In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.