Regulation of YAP translocation by myeloid Pten deficiency alleviates acute lung injury via inhibition of oxidative stress and inflammation.

BACKGROUND: Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is intricately involved in modulating the inflammatory response in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Nevertheless, the myeloid PTEN governing Hippo-YAP pathway mediated oxidative stress and inflammation in lipopolysaccharide (LPS)-induced ALI remains to be elucidate. METHODS: The floxed Pten (PtenFL/FL) and myeloid-specific Pten knockout (PtenM-KO) mice were intratracheal instill LPS (5 mg/kg) to establish ALI, then Yap siRNA mix with the mannose-conjugated polymers was used to knockdown endogenous macrophage YAP in some PtenM-KO mice before LPS challenged. The bone marrow-derived macrophages (BMMs) from PtenFL/FL and PtenM-KO mice were obtained, and BMMs were transfected with CRISPR/Cas9-mediated glycogen synthase kinase 3 Beta (GSK3ß) knockout (KO) or Yes-associated protein (YAP) KO vector subjected to LPS (100 ng/ml) challenged or then cocultured with MLE12 cells. RESULTS: Here, our findings demonstrate that myeloid-specific PTEN deficiency exerts a protective against LPS-induced oxidative stress and inflammation dysregulated in ALI model. Moreover, ablation of the PTEN-YAP axis in macrophages results in reduced nuclear factor-E2-related factor-2 (NRF2) expression, a decrease in antioxidant gene expression, augmented levels of free radicals, lipid and protein peroxidation, heightened generation of pro-inflammatory cytokines, ultimately leading to increased apoptosis in MLE12 cells. Mechanistically, it is noteworthy that the deletion of myeloid PTEN promotes YAP translocation and regulates NRF2 expression, alleviating LPS-induced ALI via the inhibition of GSK3ß and MST1 binding. CONCLUSIONS: Our study underscores the crucial role of the myeloid PTEN-YAP-NRF2 axis in governing oxidative stress and inflammation dysregulated in ALI, indicating its potential as a therapeutic target for ALI.

hucMSC-sEVs-Derived 14-3-3ζ Serves as a Bridge between YAP and Autophagy in Diabetic Kidney Disease.

As nanoscale membranous vesicles, human umbilical cord mesenchymal stem cell-derived small extracellular vesicles (hucMSC-sEVs) have attracted extensive attention in the field of tissue regeneration. Under the premise that the mechanisms of hucMSC-sEVs on the treatment of diabetic kidney disease (DKD) have not been revealed clearly, we constructed DKD rat model with success. After tail vein injection, hucMSC-sEVs effectively reduced blood glucose, maintained body weight and improved renal function in DKD rats. Notably, we found that hucMSC-sEVs suppressed YAP expression in renal cortical regions. Further in vitro experiments, we confirmed that the expression of YAP in the nucleus of renal podocytes was increased, and the level of autophagy was inhibited in the high-glucose environment, which could be reversed by intervention with hucMSC-sEVs. We screened out the key protein 14-3-3ζ, which could not only promote YAP cytoplasmic retention instead of entering the nucleus, but also enhance the level of autophagy in the cytoplasm. Ultimately, excessive YAP protein was removed by autophagy, a classic way of protein degradation. In conclusion, our study provides new strategies for the prevention of DKD and proposes the possibility of hucMSC-sEVs becoming a new treatment for DKD in the future.

Participation of Increased Circulating MAIT Cells in Lung Cancer: a Pilot Study.

Mucosal-associated invariant T (MAIT) cells are a subset of innate-like T cells that regulate the immune response via rapidly releasing inflammatory factors, including during progression of some tumors. However, the immunological role of MAIT cells is still unclear in lung cancer. We measured percentage, partial function, and clinical correlation of circulating MAIT cells from lung cancer patients through flow cytometry. Lung cancer patients displayed a high concentration of CD4+, CD8+, and activated CD38+CD8+MAIT cells, and a decrease of PD1+ double negative (DN) MAIT cells in peripheral blood. Meanwhile, increased levels of interferon-γ, interleukin (IL)-6, and 8 were examined in the serum of lung cancer patients. Importantly, we discovered a statistically positive association between accumulation of CD38+CD8+MAIT cells and reduced progression-free survival of lung cancer patients. While preliminary, the altering frequency of MAIT cells might be involved in dysfunctional immune response in lung cancer.

[Role of miR-145-5p Targeting ARK5 in Regulating the Proliferation and Apoptosis of Human Epithelial Ovarian Cancer Cells].

Objective To explore the effect of miR-145-5p on the proliferation and apoptosis of human ovarian cancer cells and the possible molecular mechanisms involved.Methods Real-time quantitative PCR was performed to detect the expression of miR-145-5p in ovarian epithelial cells and ovarian cancer cells.CCK-8 and flow cytometry were used to detect the effects of miR-145-5p overexpression on the proliferation and apoptosis of ovarian cancer cells.TargetScan was employed to predict the target genes of miR-145-5p.Western blotting,dual luciferase reporter assay and rescue experiment were employed to predict and verify the underlying molecular mechanism of miR-145-5p function.Results The expression of miR-145-5p in ovarian cancer cells was significantly lower than that in normal ovarian epithelial cells(t=4.345,P=0.049).Compared with the control group,the overexpression of miR-145-5p reduced the proliferation rate(t=-15.790,P<0.001)and increased the apoptosis rate(t=5.433,P=0.032)of ovarian cancer cells.ARK5 was predicted as the direct target gene of miR-145-5p(t=4.583,P=0.010).The cells with ARK5 overexpression showed increased proliferation rate(t=27.290,P<0.001)and decreased apoptosis rate(t=-8.241,P=0.001).The overexpression of miR-145-5p can down-regulate the mRNA(t=-12.824,P<0.001)and protein(t=-4.792,P=0.001)levels of ARK5.The rescuing expression of ARK5 significantly offset the inhibitory effects of miR-145-5p on cell proliferation(t=15.580,P=0.004)and apoptosis(t=-12.470,P=0.006).Conclusion miR-145-5p may inhibit the proliferation and promote the apoptosis of ovarian cancer cells by targeting ARK5.

Preoperative prolonged fasting causes severe metabolic acidosis: A case report.

RATIONALE: Preoperative prolonged fasting may cause starvation ketoacidosis. Herein, we report of a case of starvation ketoacidosis due to long-term fasting before surgery. PATIENT CONCERNS: We report of a case of metabolic acidosis due to prolonged fasting in a previously healthy 44-year-old woman during a total laparoscopic hysterectomy. Hyperventilation was observed to occur when the surgery was completed. Metabolic acidosis and hypoglycemia were demonstrated by blood gas analysis of the radial artery. DIAGNOSIS: Metabolic acidosis. INTERVENTIONS: The patient received sodium bicarbonate and 5% glucose fluid at the end of the surgery. OUTCOMES: The tracheal tube was successfully removed when the tidal volume of the patient returned to normal after the therapy. However, the patient suffered pulmonary edema when she was transferred to the intensive care unit (ICU). With treatments with furosemide and sodium bicarbonate, acidosis and pulmonary edema were completely corrected at 8 hours after the surgery. On the second day after the surgery, the patient suffered nausea and vomiting. Nausea and vomiting were not completely relieved on the sixth day after the operation; therefore, the patient was transferred to the Department of Gastroenterology for further therapy. LESSONS: This case suggests that although the concept of enhanced recovery after surgery (ERAS) has been adopted by most physicians because of its positive outcomes, the issue of prolonged fasting still exists, and such patients may be exposed to the risk of starvation ketoacidosis.