Integrated Single Cell and Bulk RNA-Seq Analysis Revealed Immunomodulatory Effects of Ulinastatin in Sepsis: A Multicenter Cohort Study.

Sepsis is a leading cause of morbidity and mortality in the intensive care unit, which is caused by unregulated inflammatory response leading to organ injuries. Ulinastatin (UTI), an immunomodulatory agent, is widely used in clinical practice and is associated with improved outcomes in sepsis. But its underlying mechanisms are largely unknown. Our study integrated bulk and single cell RNA-seq data to systematically explore the potential mechanisms of the effects of UTI in sepsis. After adjusting for potential confounders in the negative binomial regression model, there were more genes being downregulated than being upregulated in the UTI group. These down-regulated genes were enriched in the neutrophil involved immunity such as neutrophil activation and degranulation, indicating the immunomodulatory effects of UTI is mediated via regulation of neutrophil activity. By deconvoluting the bulk RNA-seq samples to obtain fractions of cell types, the Myeloid-derived suppressor cells (MDSC) were significantly expanded in the UTI treated samples. Further cell-cell communication analysis revealed some signaling pathways such as ANEEXIN, GRN and RESISTIN that might be involved in the immunomodulatory effects of UTI. The study provides a comprehensive reference map of transcriptional states of sepsis treated with UTI, as well as a general framework for studying UTI-related mechanisms.

miR­505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT­NFκB pathway in NSCLC cells.

MicroRNAs (miRNAs) are short non­coding RNAs, which generally regulate gene expression at the post­transcriptional level. Dysregulation of miRNAs has been reported in numerous cancer types, including lung cancer. In the present study, the role of miR­505 in non­small cell lung cancer (NSCLC) cells was investigated. miR­505 served a tumor suppressor role in NSCLC cells. By reverse transcriptase­quantitative polymerase chain reaction detection, it was demonstrated that miR­505 was downregulated in NSCLC tissues and cell lines, which is negatively associated with large tumor size, Tumor­Node­Metastasis stage and distant metastasis in patients with NSCLC. Functional studies revealed that miR­505 inhibited cell proliferation, migration, invasion and epithelial­mesenchymal transition progress in vitro and tumor growth in vivo. Mechanically, mitogen­activated protein kinase kinase kinase 3 (MAP3K3) was identified as a direct target of miR­505 by binding to its 3'untranslated region and demonstrated to mediate the tumor suppressor roles of miR­505 in NSCLC cells. The effect of miR­505 on the activation of AKT/nuclear factor­κB (NFκB) pathway, which was downstream targets of MAP3K3, was further analyzed by western blot analysis and immunofluorescence analyses. The data demonstrated the inhibition of the AKT/NFκB pathway upon overexpressing miR­505 and the activation of AKT/NFκB pathway upon silencing miR­505. Collectively, the data revealed the novel role and target of miR­505 in NSCLC cells, which may provide novel insights regarding its role in the carcinogenesis of NSCLC and its potential values for clinical applications.