Probe-based confocal laser endomicroscopy for evaluating high-altitude pulmonary edema in canines.

Background: Real-time assessment of high-altitude pulmonary edema (HAPE) remains a challenge. Probe-based confocal laser microscopy (pCLE) allows a real-time in vivo visualization of the alveoli. This study aimed to develop a new non-invasive method for analyzing microscopic images in a canine model of HAPE using pCLE. Materials and methods: This was a prospective, controlled animal study in adult male beagle dogs randomized to control and HAPE groups. The HAPE group was exposed to a high altitude of 6000 m for 48 h. The blood gas levels, lung morphological changes, infectious factors, and lung wet-to-dry ratio were analyzed in different groups. The pCLE images were described based on the volume air index (VAI), which applies an integral over specific signal intensities. Results: The lung wet-to-dry weight ratio and injury scores in the HAPE group were significantly increased compared with those of the control group. The levels of infectious factors interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-6 were significantly increased in the HAPE group compared with those in the control group. VAI was significantly decreased in the HAPE group. Conclusion: pCLE is a potential adjudicative bronchoscopic imaging technique for assessing HAPE. VAI may be acquired from quantitative parameters in the analysis of images.

CIRC_0026466 KNOCKDOWN PROTECTS HUMAN BRONCHIAL EPITHELIAL CELLS FROM CIGARETTE SMOKE EXTRACT-INDUCED INJURY BY PROMOTING THE MIR-153-3P/TRAF6/NF-ΚB PATHWAY.

ABSTRACT: Background: Considerable data have shown that circular RNAs (circRNAs) mediate the pathogenesis of chronic obstructive pulmonary disease (COPD). The study aims to analyze the function and mechanism of circ_0026466 in COPD. Methods: Human bronchial epithelial cells (16HBE) were treated with cigarette smoke extract (CSE) to establish a COPD cell model. Quantitative real-time polymerase chain reaction and Western blot were used to detect the expression of circ_0026466, microRNA-153-3p (miR-153-3p), TNF receptor associated factor 6 (TRAF6), cell apoptosis-related proteins, and NF-κB pathway-related proteins. Cell viability, proliferation, apoptosis, and inflammation were investigated by cell counting kit-8, EdU assay, flow cytometry, and enzyme-linked immunosorbent assay, respectively. Oxidative stress was evaluated by lipid peroxidation malondialdehyde assay kit and superoxide dismutase activity assay kit. The interaction between miR-153-3p and circ_0026466 or TRAF6 was confirmed by dual-luciferase reporter assay and RNA pull-down assay. Results: Circ_0026466 and TRAF6 expression were significantly increased, but miR-153-3p was decreased in the blood samples of smokers with COPD and CSE-induced 16HBE cells when compared with controls. CSE treatment inhibited the viability and proliferation of 16HBE cells but induced cell apoptosis, inflammation, and oxidative stress, but these effects were attenuated after circ_0026466 knockdown. Circ_0026466 interacted with miR-153-3p and regulated CSE-caused 16HBE cell damage by targeting miR-153-3p. Additionally, TRAF6, a target gene of miR-153-3p, regulated CSE-induced 16HBE cell injury by combining with miR-153-3p. Importantly, circ_0026466 activated NF-κB pathway by targeting the miR-153-3p/TRAF6 axis. Conclusion: Circ_0026466 absence protected against CSE-triggered 16HBE cell injury by activating the miR-153-3p/TRAF6/NF-κB pathway, providing a potential therapeutic target for COPD.

Peroxidase is a novel potential marker in glioblastoma through bioinformatics method and experimental validation.

Peroxidase (PXDN), a specific extracellular matrix (ECM)-associated protein, has been determined as a tumor indicator and therapeutic target in various tumors. However, the effects of PXDN in prognostic performance and clinical implications in glioblastoma multiforme (GBM) remains unknown. Here, we assessed PXDN expression pattern and its performance on prognosis among GBM cases from TCGA and CGGA databases. PXDN was up-regulated within GBM samples in comparison with normal control. High PXDN expression was a dismal prognostic indicator in GBM. Single cell RNA analysis was conducted to detect the cell localization of PXDN. We also set up a PPI network to explore the interacting protein associated with PXDN, including TSKU, COL4A1 and COL5A1. Consistently, functional enrichment analysis revealed that several cancer hallmarks were enriched in the GBM cases with high PXDN expression, such as epithelial-mesenchymal transition (EMT), fatty acid metabolism, glycolysis, hypoxia, inflammatory response, and Wnt/beta-catenin signaling pathway. Next, this study analyzed the association of PXDN expression and immunocyte infiltration. PXDN expression was in direct proportion to the infiltrating degrees of NK cells resting, T cells regulatory, M0 macrophage, monocytes and eosinophils. The roles of PXDN on immunity were further estimated by PXDN-associated immunomodulators. In addition, four prognosis-related lncRNAs co-expressed with PXDN were identified. Finally, we observed that PXDN depletion inhibits GBM cell proliferation and migration by in vitro experiments. Our data suggested that PXDN has the potential to be a powerful prognostic biomarker, which might offer a basis for developing therapeutic targets for GBM.

Soybean Oil Bodies as a Milk Fat Substitute Improves Quality, Antioxidant and Digestive Properties of Yogurt.

In this experiment, the effect of replacing milk fat with soybean fat body (25%, 50%, 75%, 100%) on the quality, antioxidant capacity and in vitro digestive characteristics of yogurt was investigated while maintaining the total fat content of the yogurt unchanged. The results showed that increasing the substitution amount of soy fat body for milk fat had little effect on the pH and acidity of yogurt during the storage period, while the physicochemical properties, degree of protein gel network crosslinking, saturated fatty acid content, PV value and TBARS value of the yogurt significantly decreased (p < 0.05). Meanwhile, protein content, solids content, unsaturated fatty acid content, tocopherol content and water holding capacity significantly increased (p < 0.05). Flavor analysis revealed that yogurts with soybean oil bodies were significantly different when compared to those without soybean oil bodies (p < 0.05), and yogurt with 25% substitution had the highest sensory score. After in vitro digestion, the free fatty acid release, antioxidant capacity and protein digestibility of soybean oil body yogurt were significantly higher (p < 0.05). The SDS-PAGE results showed that the protein hydrolysis of the soybean oil body yogurt was faster. Therefore, the use of an appropriate amount of soybean oil bodies to replace milk fat is able to enhance the taste of yogurt and improve the quality of the yogurt.

Synthesis and discovery of a drug candidate for treatment of idiopathic pulmonary fibrosis through inhibition of TGF-ß1 pathway.

In this study, anti-IPF lead compounds 42 and 44, derived from natural sesquiterpene lactones Isoalantolactone and alantolactone, were discovered by screening from a high-throughput TGF-ß1 reporter luciferase assay. Notably, they could reduce the myofibroblast activation and extracellular matrix deposition both in vitro and in vivo. Additionally, compounds 42 and 44 could significantly attenuate bleomycin-induced pulmonary fibrosis in mice. Further validation of pharmacokinetics study and toxicity evaluation indicated that compound 44 might be a promising anti-IPF drug candidate.

[The efficiency of macrophage extracellular trap formation induced by different inducers in vitro].

OBJECTIVE: To compare the different methods of inducing the formation of macrophage extracellular trap (MET) in vitro. METHODS: MET release was initiated by culturing RAW264.7 cells with 0.5, 1, 5, 10 µg/mL lipopolysaccharide (LPS), or 10, 25, 50, 80 µmol/L phorbolmyristate acetate (PMA), or 50, 100, 150 µg/mL silicon dioxide (SiO2). Three and 6 hours later, MET were validated by immunofluorescence staining, followed by immunofluorescence-based semi-quantitative analysis. RESULTS: Immunofluorescence staining showed that the network structures were mainly composed of DNA and histones. RAW264.7 cells treated with 1 µg/mL LPS for 6 hours produced the highest percent of MET [(37.04±10.02)%], which was statistically higher compared with control group [(7.90±2.71)%]. RAW264.7 cells treated with 80 µmol/L PMA for 6 hours also produced the higher percent of MET [(22.40±1.83)%] compared with control group [(10.11±1.13)%]. However, there was no significantly increased MET formation in cells treated with SiO2 compared with control group. CONCLUSION: LPS and PMA can induce MET formation in vitro, while SiO2 was not efficient inducer.