High-content analysis identified synergistic drug interactions between INK128, an mTOR inhibitor, and HDAC inhibitors in a non-small cell lung cancer cell line.

BACKGROUND: The development of drug resistance is a major cause of cancer therapy failures. To inhibit drug resistance, multiple drugs are often treated together as a combinatorial therapy. In particular, synergistic drug combinations, which kill cancer cells at a lower concentration, guarantee a better prognosis and fewer side effects in cancer patients. Many studies have sought out synergistic combinations by small-scale function-based targeted growth assays or large-scale nontargeted growth assays, but their discoveries are always challenging due to technical problems such as a large number of possible test combinations. METHODS: To address this issue, we carried out a medium-scale optical drug synergy screening in a non-small cell lung cancer cell line and further investigated individual drug interactions in combination drug responses by high-content image analysis. Optical high-content analysis of cellular responses has recently attracted much interest in the field of drug discovery, functional genomics, and toxicology. Here, we adopted a similar approach to study combinatorial drug responses. RESULTS: By examining all possible combinations of 12 drug compounds in 6 different drug classes, such as mTOR inhibitors, HDAC inhibitors, HSP90 inhibitors, MT inhibitors, DNA inhibitors, and proteasome inhibitors, we successfully identified synergism between INK128, an mTOR inhibitor, and HDAC inhibitors, which has also been reported elsewhere. Our high-content analysis further showed that HDAC inhibitors, HSP90 inhibitors, and proteasome inhibitors played a dominant role in combinatorial drug responses when they were mixed with MT inhibitors, DNA inhibitors, or mTOR inhibitors, suggesting that recessive drugs could be less prioritized as components of multidrug cocktails. CONCLUSIONS: In conclusion, our optical drug screening platform efficiently identified synergistic drug combinations in a non-small cell lung cancer cell line, and our high-content analysis further revealed how individual drugs in the drug mix interact with each other to generate combinatorial drug response.

Insight into the lncRNA-mRNA Co-Expression Profile and ceRNA Network in Lipopolysaccharide-Induced Acute Lung Injury.

Long non-coding RNAs (lncRNAs) participate in acute lung injury (ALI). However, their latent biological function and molecular mechanism have not been fully understood. In the present study, the global expression profiles of lncRNAs and mRNAs between the control and lipopolysaccharide (LPS)-stimulated groups of human normal lung epithelial cells (BEAS-2B) were determined using high-throughput sequencing. Overall, a total of 433 lncRNAs and 183 mRNAs were differentially expressed. A lncRNA-mRNA co-expression network was established, and then the top 10 lncRNAs were screened using topological methods. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis results showed that the key lncRNAs targeting mRNAs were mostly enriched in the inflammatory-related biological processes. Gene set variation analysis and Pearson's correlation coefficients confirmed the close correlation for the top 10 lncRNAs with inflammatory responses. A protein-protein interaction network analysis was conducted based on the key lncRNAs targeting mRNAs, where IL-1ß, IL-6, and CXCL8 were regarded as the hub genes. A competing endogenous RNA (ceRNA) modulatory network was created with five lncRNAs, thirteen microRNAs, and twelve mRNAs. Finally, real-time quantitative reverse transcription-polymerase chain reaction was employed to verify the expression levels of several key lncRNAs in BEAS-2B cells and human serum samples.

Inhibition of gp130 alleviates LPS-induced lung injury by attenuating apoptosis and inflammation through JAK1/STAT3 signaling pathway.

BACKGROUND AND OBJECTIVE: Acute lung injury or acute respiratory distress syndrome (ALI/ARDS) is a life-threatening respiratory disease. Gp130 is a signal transduction receptor that participates in a variety of essential biological processes. The biological function of gp130 in ALI/ARDS is unclear. This study aims to investigate the roles and potential mechanisms of gp130 in lung injury induced by lipopolysaccharide (LPS). METHODS: The ALI/ARDS mouse model was established using intratracheal LPS administration. Hematoxylin and eosin staining and bronchoalveolar lavage fluid analysis were used to evaluate the degree of lung injury. Cell apoptosis was assessed by TUNEL staining, flow cytometry, and western blot. Then the expression of gp130, IL-6, IL-10, TNF-α, and the JAK1/STAT3 signaling pathway-related proteins was assessed by RT-PCR, western blot, and immunohistochemistry. RESULTS: The expression of gp130 increased after 24 h of LPS treatment. Inhibiting gp130 improved inflammatory infiltration and alveolar collapsed, decreased IL-6 and TNF-α levels, raised IL-10 levels, and decreased cell apoptosis in LPS-induced mice. Meanwhile, suppressing gp130 reduced the inflammatory response and cell apoptosis in LPS-induced Beas-2B cells. Furthermore, p-JAK1 and p-STAT3 expressions were elevated after LPS stimulation and decreased following gp130 inhibition, suggesting that gp130 may regulate the JAK1/STAT3 signaling pathway in LPS-induced mice and Beas-2B cells. CONCLUSION: The findings suggest that gp130 regulates the inflammatory response and cell apoptosis through the JAK1/STAT3 signaling pathway, thereby mitigating LPS-induced lung injury. Gp130 may be a potential therapeutic target for ALI/ARDS.

SIRT6 ameliorates LPS-induced apoptosis and tight junction injury in ARDS through the ERK1/2 pathway and autophagy.

Sirtuin6 (SIRT6) has been demonstrated to be involved in a range of physiological processes and diseases, while its role in acute respiratory distress syndrome (ARDS) remains unclear. Therefore, this study focused on the role and underlying mechanism of SIRT6 in ARDS with the aim of identifying potential therapeutic targets. In this study, we found that SIRT6 was significantly decreased in lipopolysaccharide (LPS)-induced A549 cells and a murine model. In vitro overexpression of SIRT6 restored the expression of tight junction proteins (ZO-1 and occludin) and alleviated cell apoptosis and inflammation, while knockdown of SIRT6 aggravated the loss of tight junction proteins (ZO-1 and occludin) and promoted cell apoptosis and inflammation in LPS-induced A549 cells. Furthermore, the overexpression of SIRT6 enhanced autophagy and inhibited the ERK1/2 pathway, while the knockdown of SIRT6 inhibited autophagy and activated the ERK1/2 pathway. The autophagy activator rapamycin and the ERK1/2 inhibitor PD98059 rescued the effects of SIRT6 knockdown on tight junction proteins, apoptosis, and inflammation. Mechanistically, SIRT6 deacetylated histone 3 at Lys9 to negatively regulate the ERK1/2 pathway. In vivo, the SIRT6-specific inhibitor OSS_128167 also significantly accelerated LPS-induced loss of tight junction proteins, lung inflammation, and apoptosis. Meanwhile, the SIRT6-specific inhibitor OSS_128167 also activated the ERK1/2 pathway and inhibited lung autophagy. These results suggested that SIRT6 could ameliorate the loss of tight junction proteins, inflammation, and apoptosis in LPS-induced ARDS by inhibiting the ERK1/ 2 pathway and enhancing autophagy, indicating that SIRT6 plays a beneficial role in ARDS and might be a potential therapeutic target for ARDS.

Clinical characteristics of allergic bronchopulmonary aspergillosis in patients with and without bronchiectasis.

OBJECTIVE: Allergic bronchopulmonary aspergillosis (ABPA) is classified radiologically as serologic ABPA (ABPA-S) or ABPA with central bronchiectasis (ABPA-CB). This retrospective case series study aimed to describe and compare the clinical characteristics of both forms of ABPA. METHODS: Patients with ABPA treated in the hospital between February 2011 and June 2019 were enrolled and were divided into ABPA-S and ABPA-CB groups based on whether their cases were complicated with central bronchiectasis. Demographic data, symptoms, laboratory values, comorbidities, and image findings were collected. ABPA-S patients were followed up retrospectively through medical records. RESULTS: Ninety-three (93) patients were enrolled, including 74 ABPA-CB patients and 19 ABPA-S patients. The most common predisposing condition was asthma (36.6%), with a median course of 30 years (IQR 13-42.5) prior to ABPA diagnosis. Patients of 54.8% had been misdiagnosed, with ABPA-S more likely than ABPA-CB to have been misdiagnosed as asthma (p < 0.01). Obstructive ventilation dysfunction and mixed ventilation dysfunction were found in 21 patients (22.6%) and 16 patients (17.2%), respectively. Compared with ABPA-S, ABPA-CB had a higher median blood eosinophil count (880 vs. 700 cells/µl), serum IgE (2957 vs. 2616 IU/ml), and Aspergillus fumigatus specific-IgE (20.6 vs. 7.31 kUA/L), although these findings were not statistically significant. Three ABPA-S patients developed bronchiectasis during follow-up and experienced relapses more than twice. CONCLUSIONS: Our findings suggested that the clinical characteristics between ABPA-CB and ABPA-S were mostly similar. ABPA-S had a relatively lower immunological activity level than ABPA-CB but was still immunologically active and could develop bronchiectasis.

Molecular Evolution and Local Root Heterogeneous Expression of the Chenopodium quinoa ARF Genes Provide Insights into the Adaptive Domestication of Crops in Complex Environments.

Auxin response factors (ARFs) influence plant growth and development via the coupling of basic biological processes. However, the evolution, expansion, and regulatory mechanisms of ARFs in the domesticated crop quinoa after artificial selection remain elusive. In this study, we systematically identified 30 Chenopodium quinoa ARFs (CqARFs). In this typical domesticated crop, ARFs divided into three subfamilies are subjected to strong purification selection and have a highly conserved evolutionary pattern. Polyploidy is the primary reason for the expansion of the ARF family after quinoa domestication. The expression patterns of CqARFs in different tissues have been differentiated, and CqARF2, 5, 9 and 10 from class A have the characteristics of local heterogeneous expression in different regions of roots, which may be the key factors for crops to respond in complex environments. Overall, we examined the evolution and expansion of ARFs in representative domesticated crops using the genome, transcriptome, and molecular biology and discovered a class A ARF-centered heterogeneous expression network that played an important role in auxin signaling and environmental responses. We provide new insights into how ARFs promote domesticated crop adaptation to artificial selection by polyploid expansion.

Ameliorative effects of eosinophil deficiency on immune response, endoplasmic reticulum stress, apoptosis, and autophagy in fungus-induced allergic lung inflammation.

BACKGROUND: Respiratory fungal exposure is known to be associated with various allergic pulmonary disorders. Eosinophils have been implicated in tissue homeostasis of allergic inflammation as both destructive effector cells and immune regulators. What contributions eosinophils have in Aspergillus fumigatus (Af)-induced allergic lung inflammation is worthy of investigating. METHODS: We established the Af-exposed animal asthmatic model using eosinophil-deficient mice, ∆dblGATA1 mice. Airway inflammation was assessed by histopathological examination and total cell count of bronchoalveolar lavage fluid (BALF). The protein level in BALF and lung mRNA level of type 2 cytokines IL-4, IL-5, and IL-13 were detected by ELISA and qRT-PCR. We further studied the involvement of endoplasmic reticulum (ER) stress, apoptosis, and autophagy by western blots, qRT-PCR, immunofluorescence, TUNEL, or immunohistochemistry. RNA-Seq analysis was utilized to analyze the whole transcriptome of Af-exposed ∆dblGATA1 mice. RESULTS: Hematoxylin and eosin (HE) staining and periodic acid-Schiff staining (PAS) showed that airway inflammation and mucus production were alleviated in Af-challenged ∆dblGATA1 mice compared with wild-type controls. The protein and mRNA expressions of IL-4, IL-5, and IL-13 were reduced in the BALF and lung tissues in Af-exposed ∆dblGATA1 mice. The results demonstrated that the significantly increased ER stress markers (GRP78 and CHOP) and apoptosis executioner caspase proteases (cleaved caspase-3 and cleaved caspase-7) in Af-exposed wild-type mice were all downregulated remarkably in the lungs of ∆dblGATA1 mice with Af challenge. In addition, the lung autophagy in Af-exposed ∆dblGATA1 mice was found elevated partially, manifesting as higher expression of LC3-II/LC3-I and beclin1, lower p62, and downregulated Akt/mTOR pathway compared with Af-exposed wild-type mice. Additionally, lung RNA-seq analysis of Af-exposed ∆dblGATA1 mice showed that biological processes about chemotaxis of lymphocytes, neutrophils, or eosinophils were enriched but without statistical significance. CONCLUSIONS: In summary, eosinophils play an essential role in the pathogenesis of Af-exposed allergic lung inflammation, whose deficiency may have relation to the attenuation of type 2 immune response, alleviation of ER stress and apoptosis, and increase of autophagy. These findings suggest that anti-eosinophils therapy may provide a promising direction for fungal-induced allergic pulmonary diseases.

Reversible bronchial dilatation in adults.

Bronchiectasis is characterized by the destruction of bronchial wall and persistent irreversible bronchiectasis due to respiratory infection and bronchial obstruction. Reversible bronchiectasis is rarely reported in adults. The study aims to evaluate dynamic changes in chest computed tomography (CT) findings and relevant factors affecting improvement or reversal of bronchiectasis. A total of 239 patients with bronchiectasis admitted to our hospital from January 2009 to December 2019 were retrospectively analyzed. 23 patients (group A) with bronchiectasis reversion or improvement confirmed by chest CT were matched with patients in group B according to gender and age (difference <5 years). 23 patients (group B) with bronchiectasis progression or no obvious improvement confirmed by chest CT were selected from the remaining patients. The clinical features and imaging findings of the two groups were compared. The duration of disease in group A was less than that in group B, and the stable period was longer (p < 0.05). There was no significant difference in other clinical features between the two groups. In contrast to the traditional concept of "Bronchiectasis", our results suggested that some adult bronchiectasis can be improved and completely radiologically reversed.

Gastric floating tablet improves the bioavailability and reduces the hypokalemia effect of gossypol in vivo.

Gossypol (Gos) is a natural polyphenolic compound that has shown a number of valuable biological properties such as antifertility, antioxidation, and antitumor activities. However, the clinical application of Gos has been hindered by its notable adverse effects such as hypokalemia, hemolytic anemia, and so on. Using sustained-release dosage form provides a hopeful solution to this problem. In this study, a gastric floating tablet for sustained-release of Gos (Gos-GFT) was developed using polyvinylpyrrolidone, hydroxypropyl methyl cellulose, ethyl cellulose, lactose, sodium bicarbonate, and magnesium stearate. Gos-GFT had an average weight of around 200 mg with a drug content percentage of around 13.66%. The physicochemical properties of Gos-GFT satisfied the pharmacopoeial requirements for tablets. Gos-GFT was able to float in an acidic medium and had a sustained drug release for over 12 h. In vivo studies showed that the relative bioavailability of Gos-GFT, as compared with Gos powders, was larger than that of a non-gastric floating tablet which was a dosage form used for comparison with Gos-GFT. Furthermore, compared with the Gos powders and the non-gastric floating Gos tablets, Gos-GFT could prolong the in vivo action time of Gos, and significantly relieve hypokalemia which is a major adverse effect of Gos. These properties made Gos-GFT a promising Gos preparation that warrants further investigation for more extensive clinical applications of this natural compound.

The association between serum albumin/prealbumin level and disease severity in non-CF bronchiectasis.

Non-cystic fibrosis (non-CF) bronchiectasis is a chronic pulmonary disease that can lead to malnutrition. Serum prealbumin and albumin level are related to inflammatory and nutritional status. Thus, we aimed to confirm our hypothesis that low serum albumin and prealbumin level, as well as body mass index (BMI), is correlated to severe non-CF bronchiectasis. We conducted a retrospective cross-sectional study of 128 patients, including 75 patients with prealbumin test and 79 patients with albumin test. Detailed medical history was recorded, including pulmonary function tests and high-resolution computed tomography. bronchiectasis severity index (BSI) and FACED scores were calculated. Leicester Cough Questionnaire, Quality of Life Questionnaire-Bronchiectasis, chronic obstructive pulmonary disease (COPD) assessment test and Patient Health Questionnaire-9 questionnaires were used to assess patients' clinical symptoms. Correlation analysis showed that BSI score was more correlated to patients' clinical symptoms than FACED. Thus, patients were divided into three groups of different severity based on BSI score. Albumin, prealbumin and BMI showed a significant difference between three groups. Correlation and multivariable linear regression analysis showed that serum albumin and prealbumin level were correlated to BSI, FACED and questionnaires. The analysis between three indices and PFT/high-resolution computed tomography (HRCT) showed that prealbumin, albumin and BMI could reflect the PFT and modified Reiff score in non-CF bronchiectasis. In conclusion, BMI, albumin and prealbumin showed a significant correlation with the BSI, FACED, as well as patients' clinical symptoms. Among them, serum albumin was the indicator most strongly associated with the BSI and questionnaires, while prealbumin could better reflect lung function decline and radiological severity.

Suppressing Endoplasmic Reticulum Stress Alleviates LPS-Induced Acute Lung Injury via Inhibiting Inflammation and Ferroptosis.

Acute lung injury (ALI) is a life-threatening clinical disorder with high mortality rate. Ferroptosis is a new type of programmed cell death with lipid peroxidation and iron ion overloading as the main characteristics. Endoplasmic reticulum (ER) stress and ferroptosis play pivotal roles in the pathogenesis of ALI. The study aimed to investigate the underlying relationship between ER stress and ferroptosis in ALI. The ER stress inhibitor 4-phenylbutyric acid (4-PBA) alleviated LPS-induced inflammation, and decreased IL-1ß, IL-6, and TNF-α levels in BALF and lungs. The increased MDA and decreased GSH induced by LPS were partially reversed by 4-PBA, which also inhibited the expressions of ferroptosis-related protein ACSL4, COX-2, and FTH1. TEM further confirmed the ferroptosis within airway epithelia cells was ameliorated by 4-PBA. Moreover, 4-PBA reduced the production of ROS and lipid ROS in LPS-exposed BEAS-2B cells in a concentration-dependent way. Meanwhile, 4-PBA mitigated LPS-induced cell apoptosis in vivo and in vitro. Mechanistically, the MAPK signaling pathway activated by LPS was downregulated by 4-PBA. Collectively, these findings suggested that 4-PBA protected against ALI by inhibiting inflammation and ferroptosis through downregulating ER stress, thus providing a potential intervention for ALI and revealing the possible interaction between ER stress and ferroptosis in ALI.

Smart Dual-Exsolved Self-Assembled Anode Enables Efficient and Robust Methane-Fueled Solid Oxide Fuel Cells.

Perovskite oxides have emerged as alternative anode materials for hydrocarbon-fueled solid oxide fuel cells (SOFCs). Nevertheless, the sluggish kinetics for hydrocarbon conversion hinder their commercial applications. Herein, a novel dual-exsolved self-assembled anode for CH4 -fueled SOFCs is developed. The designed Ru@Ru-Sr2 Fe1.5 Mo0.5 O6-δ (SFM)/Ru-Gd0.1 Ce0.9 O2-δ (GDC) anode exhibits a unique hierarchical structure of nano-heterointerfaces exsolved on submicron skeletons. As a result, the Ru@Ru-SFM/Ru-GDC anode-based single cell achieves high peak power densities of 1.03 and 0.63 W cm-2 at 800 °C under humidified H2 and CH4 , surpassing most reported perovskite-based anodes. Moreover, this anode demonstrates negligible degradation over 200 h in humidified CH4 , indicating high resistance to carbon deposition. Density functional theory calculations reveal that the created metal-oxide heterointerfaces of Ru@Ru-SFM and Ru@Ru-GDC have higher intrinsic activities for CH4 conversion compared to pristine SFM. These findings highlight a viable design of the dual-exsolved self-assembled anode for efficient and robust hydrocarbon-fueled SOFCs.

Partial Exsolution Enables Superior Bifunctionality of Ir@SrIrO3 for Acidic Overall Water Splitting.

The pursuit of efficient and durable bifunctional electrocatalysts for overall water splitting in acidic media is highly desirable, albeit challenging. SrIrO3 based perovskites are electrochemically active for oxygen evolution reaction (OER), however, their inert activities toward hydrogen evolution reaction (HER) severely restrict the practical implementation in overall water splitting. Herein, an Ir@SrIrO3 heterojunction is newly developed by a partial exsolution approach, ensuring strong metal-support interaction for OER and HER. Notably, the Ir@SrIrO3-175 electrocatalyst, prepared by annealing SrIrO3 in 5% H2 atmosphere at 175 °C, delivers ultralow overpotentials of 229 mV at 10 mA cm-2 for OER and 28 mV at 10 mA cm-2 for HER, surpassing most recently reported bifunctional electrocatalysts. Moreover, the water electrolyzer using the Ir@SrIrO3-175 bifunctional electrocatalyst demonstrates the potential application prospect with high electrochemical performance and excellent durability in acidic environment. Theoretical calculations unveil that constructing Ir@SrIrO3 heterojunction regulates interfacial electronic redistribution, ultimately enabling low energy barriers for both OER and HER.

Reverse Atom Capture on Perovskite Surface Enabling Robust and Efficient Cathode for Protonic Ceramic Fuel Cells.

Protonic ceramic fuel cells (PCFCs) hold potential for sustainable energy conversion, yet their widespread application is hindered by the sluggish kinetics and inferior stability of cathode materials. Here, a facile and efficient reverse atom capture technique is developed to manipulate the surface chemistry of PrBa0.5Sr0.5Co1.5Fe0.5O5+ δ (PBSCF) cathode for PCFCs. This method successfully captures segregated Ba and Sr cations on the PBSCF surface using W species, creating a (Ba/Sr)(Co/Fe/W)O3- δ (BSCFW)@PBSCF heterostructure. Benefiting from enhanced kinetics of proton-involved oxygen reduction reaction and strengthened chemical stability, the single cell using the optimized 2W-PBSCF cathode demonstrates an exceptional peak power density of 1.32 W cm-2 at 650 °C and maintains durable performance for 240 h. Theoretical calculations unveil that the BSCFW perovskite delivers lower oxygen vacancy formation energy, hydration energy, and proton transfer energy compared to the PBSCF perovskite. This protocol offers new insights into advanced atom capture techniques for sustainable energy infrastructures.

PAI-1 Deficiency Promotes NET-mediated Pyroptosis and Ferroptosis during Pseudomonas Aeruginosa-induced Acute Lung Injury by Regulating the PI3K/MAPK/AKT Axis.

Circulating neutrophil extracellular trap (NET) formation is an adaptive process during acute lung injury (ALI). The important role of plasminogen activator inhibitor (PAI)-1 in NET formation during ALI remains unclear. This research intends to examine the impacts of the decrease in PAI-1 levels on NET formation and the underlying mechanism. We found a relative association between the increase in plasma NET levels and thromboinflammation-induced lung damage in patients with ARDS. PAI-1 knockout (KO) mice exhibited significant increases in Pseudomonas aeruginosa (PAO1 strain)-induced ALI, inflammation, inflammatory cell accumulation, and proinflammatory cytokine secretion, and wild-type mice exhibited the opposite changes. During PAO1-induced ALI, PAI-1 KO increased NET release and the levels of prothrombotic markers in mice. PAI-1 deficiency also promoted NET formation and NET-mediated pyroptosis and ferroptosis by activating the PI3K/MAPK/AKT pathway in a PAO1-induced ALI mouse model. In conclusion, PAI-1 KO exacerbated PAO1-induced pneumonia-associated injury and contributed to NET-mediated pyroptosis and ferroptosis through PI3K/MAPK/AKT pathway activation. Thus, targeting PAI-1 and NETs may be a promising therapeutic approach for ameliorating pneumonia and thromboinflammation-associated ALI.

Nuclear SPHK2/S1P induces oxidative stress and NLRP3 inflammasome activation via promoting p53 acetylation in lipopolysaccharide-induced acute lung injury.

A bulk of evidence identified that macrophages, including resident alveolar macrophages and recruited macrophages from the blood, played an important role in the pathogenesis of acute respiratory distress syndrome (ARDS). However, the molecular mechanisms of macrophages-induced acute lung injury (ALI) by facilitating oxidative stress and inflammatory responses remain unclear. Herein, we noticed that the levels of mitochondrial reactive oxygen species (mtROS), SPHK2 and activated NLRP3 inflammasome were higher in peripheral blood mononuclear cells (PBMCs) of ARDS patients than that in healthy volunteers. Similar observations were recapitulated in LPS-treated RAW264.7 and THP-1 cells. After exposure to LPS, the SPHK2 enzymatic activity, NLRP3 inflammasome activation and mtROS were significantly upregulated in macrophages. Moreover, knockdown SPHK2 via shRNA or inhibition SPHK2 could prominently decrease LPS-induced M1 macrophage polarization, oxidative stress and NLRP3 inflammasome activation. Further study indicated that upregulated SPHK2 could increase nuclear sphingosine-1-phosphate (S1P) levels and then restrict the enzyme activity of HDACs to facilitate p53 acetylation. Acetylation of p53 reinforced its binding to the specific region of the NLRP3 promoter and drove expression of NLRP3. In the in vivo experiments, it was also observed that treating with Opaganib (ABC294640), a specific SPHK2 inhibitor, could observably alleviate LPS-induced ALI, evidencing by lowered infiltration of inflammatory cells, increased M2 macrophages polarization and reduced oxidative damage in lung tissues. Besides, SPHK2 inhibition can also decrease the accumulation of acetylated p53 protein and the activation of NLRP3 inflammasome. Taken together, our results demonstrated for the first time that nuclear S1P can regulate the acetylation levels of non-histone protein through affecting HDACs enzyme activities, linking them to oxidative stress and inflammation in response to environmental signals. These data provide a theoretical basis that SPHK2 may be an effective therapeutic target of ARDS.

Microarray profiling of circular RNAs in LPS-induced Beas-2B cells and identification of circ_2979 as a potential biomarker for ARDS.

Acute respiratory distress syndrome (ARDS) is a major cause of high mortality and morbidity in critically ill patients. Circular RNAs (CircRNAs) are widely expressed in numerous tissues and are associated with various diseases. However, the role of circRNAs in ARDS remains unclear. In this study, we found that cell viability and proliferation were reduced in lipopolysaccharide (LPS)-induced Beas-2B cells. Microarray analysis identified 1131 differentially expressed circRNAs in LPS-treated Beas-2B cells, with 623 circRNAs significantly upregulated and 508 circRNAs strongly downregulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed significant enrichment and indicated potential functions and pathways of differentially expressed circRNAs. Reverse transcription-polymerase chain reaction (RT-PCR) analysis confirmed that expression of circ_2979, circ_5438, circ_4557 and circ_2066 in LPS-induced Beas-2B cells was consistent with the results obtained by RNA sequencing (RNA-seq). Additionally, we recruited 17 patients with ARDS and 13 healthy volunteers and found that expression of circ_2979 in serum was significantly increased in the patients with ARDS compared with healthy volunteers. Spearman's analyses indicated that circ_2979 was correlated with partial pressure of carbon dioxide in arterial blood (PaCO2), the ratio of partial pressure of arterial oxygen to the fraction of inspired oxygen (PaO2/FiO2), interleukin 2 receptor (IL-2R) and fibrinogen (FIB). The results suggested that circRNAs may play an important role in the progression of ARDS, and that circ_2979 may serve as a diagnosis and prognosis biomarker for ARDS.

Identification and validation of ferroptosis-related genes in lipopolysaccharide-induced acute lung injury.

BACKGROUND: Emerging evidence reveals the important role of ferroptosis in the pathophysiological process of acute lung injury (ALI). We aimed to identify and validate the potential ferroptosis-related genes of ALI through bioinformatics analysis and experimental validation. METHODS: Murine ALI model was established via intratracheal instillation with LPS and confirmed by H&E staining and transmission electronic microscopy (TEM). RNA sequencing (RNA-seq) was used to screen differentially expressed genes (DEGs) between control and ALI model mice. The potential differentially expressed ferroptosis-related genes of ALI were identified using the limma R package. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, gene set enrichment analysis (GSEA), and protein-protein interactions (PPI) were applied for the differentially expressed ferroptosis-related genes. CIBERSORT tool was used to conduct immune cell infiltration analysis. Finally, protein expressions and RNA expression of ferroptosis DEGs were validated in vivo and in vitro by western blots and RT-qPCR. RESULTS: Among 5009 DEGs, a total of 86 differentially expressed ferroptosis-related genes (45 up-regulated genes and 41 down-regulated genes) were identified in the lungs between control and ALI. GSEA analysis showed that the genes enriched were mainly involved in response to molecule of bacterial origin and fatty acid metabolic process. The GO and KEGG enrichment analysis indicated that the top 40 ferroptosis DEGs were mainly enriched in reactive oxygen species metabolic process, HIF-1signaling pathway, lipid and atherosclerosis, and ferroptosis. The PPI results and Spearman correlation analysis suggested that these ferroptosis-related genes interacted with each other. Immune infiltration analysis confirmed that ferroptosis DEGs were closely related to immune response. Consistent with the RNA-seq data, the western blot and RT-qPCR unveiled increased mRNA expressions of Cxcl2, Il-6, Il-1ß, and Tnfα, and protein expressions of FTH1, TLR4 as well as decreased ACSL3 in LPS-induced ALI. In vitro, the upregulated mRNA levels of CXCL2, IL-6, SLC2A1, FTH1, TNFAIP3, and downregulated NQO1 and CAV1 in LPS-stimulated BEAS-2B and A549 cells were verified. CONCLUSION: We identified 86 potential ferroptosis-related genes of LPS-induced ALI through RNA-seq. Several pivotal ferroptosis-related genes involved in lipid metabolism and iron metabolism were implicated in ALI. This study may be helpful to expand our understanding of ALI and provide some potential targets to counteract ferroptosis in ALI.

Polymer Composite with Enhanced Thermal Conductivity and Insulation Properties through Aligned Al2O3 Fiber.

Thermoplastic polyolefins, such as polyethylene (PE), are traditionally one of the most widely used polymer classes with applications in the electric industry, and their nanocomposites have caught the interest of researchers. The linear filler is shown to be beneficial in decreasing the charge injection and hindering the formation of charge packs. So, we demonstrate a novel composite with excellent properties. The low-density polyethylene (LDPE) composite with aligned aluminum oxide (Al2O3) fiber has been prepared in electric field conditions. The direction of the Al2O3 fiber was parallel to the thickness direction of the LDPE composite. The breakdown strength of the Al2O3/LDPE composite with 0.2% aligned Al2O3 fiber was 498 kV/mm, which is higher than other fillers induced. The aligned Al2O3 fiber has effect on preventing accumulation of space charge and reducing the amount of free electron in the material. In addition, the thermal conductivity of the LDPE composite (0.22 W/m·K) was increased to 0.85 W/m·K when doped with 0.5 wt% aligned Al2O3 fiber. The present structure provides a new possibility for mass new nanocomposites with excellent microstructures and remarkable functionality.

Albumin level as an independent predictive factor for adverse outcomes in COVID-19 patients: a retrospective cohort study.

INTRODUCTION: Research on the association between albumin (ALB) level and clinical outcomes of coronavirus disease 2019 (COVID-19) are limited. This study aimed to investigate the relationship between albumin level at the time of admission and adverse outcomes in patients with COVID-19. METHODOLOGY: This was a retrospective cohort study with 199 COVID-19 patients from five designated hospitals in Fujian Province who were enrolled between 22 January and 27 February, 2020. Clinical characteristics and laboratory values at the time of admission were collected. Adverse outcomes were defined as meeting at least one of the following criteria: development of acute respiratory distress syndrome (ARDS), respiratory failure, shock, multiple organ failure (MOF), intensive care unit (ICU) admission and in-hospital mortality event. The univariate and multivariate linear regression models and generalized additive models (GAM) were used to analyze the relationship between ALB and adverse outcomes. RESULTS: A non-linear relationship with an inflection point of 32.6g/L was detected between ALB and adverse outcomes after adjusting for potential confounders. The odds ratio and the confidence intervals on the left and right sides of the inflection point were 0.204 (0.061-0.681) and 0.908 (0.686-1.203), respectively. This suggested that ALB was negatively correlated with adverse outcomes when ALB was less than 32.6 g/L, and for every 1 unit increase in ALB, the risk of adverse outcomes was reduced by 79.6%. CONCLUSIONS: The relationship between ALB and adverse outcomes of COVID-19 is non-linear. ALB level is an independent predictive factor for adverse outcomes in COVID-19 patients.