Rutin targets AKT to inhibit ferroptosis in ventilator-induced lung injury.

Our previous research confirmed that rutin reduced ventilator-induced lung injury (VILI) in mice. Ferroptosis has been reported to participate in the pathogenic process of VILI. We will explore whether rutin inhibits ferroptosis to alleviate VILI. A mouse model of VILI was constructed with or without rutin pretreatment to perform a multiomics analysis. Hematoxylin-eosin (HE) staining and transmission electron microscopy were used to evaluate lung injury in VILI mice. Dihydroethidium (DHE) staining and the malondialdehyde (MDA) and superoxide dismutase (SOD) levels were detected. Molecular docking was performed to determine the binding affinity between rutin and ferroptosis-related proteins. Western blot analysis, real-time PCR (RT-PCR) and immunohistochemical (IHC) staining were conducted to detect the expression levels of GPX4, XCT, ACSL4, FTH1, AKT and p-AKT in lung tissues. Microscale thermophoresis (MST) was used to evaluate the binding between rutin and AKT1. Transcriptomic and proteomic analyses showed that ferroptosis may play a key role in VILI mice. Metabolomic analysis demonstrated that rutin may affect ferroptosis via the AKT pathway. Molecular docking analysis indicated that rutin may regulate the expression of ferroptosis-related proteins. Moreover, rutin upregulated GPX4 expression and downregulated the expression of XCT, ACSL4 and FTH1 in the lung tissues. Rutin also increased the ratio of p-AKT/AKT and p-AKT expression. MST analysis showed that rutin binds to AKT1. Rutin binds to AKT to activate the AKT signaling pathway, contributing to inhibit ferroptosis, thus preventing VILI in mice. Our study elucidated a possible novel strategy of involving the use of rutin for preventing VILI.

Rutin alleviates ventilator-induced lung injury by inhibiting NLRP3 inflammasome activation.

Whether rutin relieves ventilator-induced lung injury (VILI) remains unclear. Here, we used network pharmacology, bioinformatics, and molecular docking to predict the therapeutic targets and potential mechanisms of rutin in the treatment of VILI. Subsequently, a mouse model of VILI was established to confirm the effects of rutin on VILI. HE staining showed that rutin alleviated VILI. TUNEL staining showed that rutin reduced apoptosis in the lung tissue of mice with VILI, and the same change was observed in the ratio of Bax/Bcl2. Furthermore, rutin reduced the expression of NLRP3, ASC, Caspase1, IL1ß, and IL18 in the lung tissues of mice with VILI. Mechanistically, rutin suppressed the TLR4/NF-κB-P65 pathway, which promoted the M1 to M2 macrophage transition and alleviated inflammation in mice with VILI. Rutin relieved NLRP3 inflammasome activation by regulating M1/M2 macrophage polarization and inhibiting the activation of the TLR4/NF-κB-P65 pathway, resulting in the amelioration of VILI in mice.

GCH1 reduces LPS-induced alveolar macrophage polarization and inflammation by inhibition of ferroptosis.

OBJECTIVE: GTP cyclohydrolase 1(GCH1) was reported to protect against ferroptosis. However, it is not clear whether GCH1 reduced lipopolysaccharide (LPS)-induced macrophage polarization and inflammation by inhibition of ferroptosis. METHODS: Bioinformatics analysis was used to screen differential expression genes (DEGs) and obtain the different pathways and biological features. Lasso cox regression analysis with ferroptosis related DEGs was established to screen the most relevant genes for disease risk. LPS induced Raw264.7 macrophage polarization model and GCH1-specific siRNA oligos transfection were performed to confirm the function of GCH1. Immunofluorescence staining, western blot and quantitative real-time PCR were performed to detect the expression of iNOS, CD206, GCH1, IL6, SLC2A6, F4/80, IL1ß, TNFα, IL10, GPX4, ACSL4, AMPK and p-AMPK in macrophages. The levels of ROS, SOD, MDA and GSH were detected according to the instructions of the reagent kit, respectively. RESULTS: 542 DEGs were screened from GSE40885 microarray. GO and KEGG pathway enrichment analysis showed that the upregulated DEGs induced by LPS in alveolar macrophage were closely associated with inflammatory and immune responses, the downregulated DEGs were related to lipid metabolism, insulin resistance and AMPK signal pathway. Lasso cox regression analysis screened GCH1, IL6, and SLC2A6. Our experimental results showed that the expression of GCH1 and IL6 in the LPS group was higher than that in the control group, but there was no difference in the expression of SLC2A6. Bioinformatics analysis with GSE112720 observed that ferroptosis was enriched in GCHfl/fl + LPS group compared with GCHfl/flTie2cre + LPS group and GCHfl/fl + control group. Silence of GCH1 increased the levels of IL6, TNF-α and IL-1ß and decreased IL10 level. Silence of GCH1 increased iNOS level and decreased CD206 level. Moreover, silence of GCH1 raised ferroptosis induced by LPS in macrophages and suppressed the activity of AMPK pathway. CONCLUSIONS: GCH1 inhibited ferroptosis in LPS-stimulated macrophages, reduced macrophage toward to M1 polarization and inflammatory response.

Incidence, outcomes and risk factors of barotrauma in veno-venous extracorporeal membrane oxygenation for acute respiratory distress syndrome.

BACKGROUND: Although acute respiratory distress syndrome (ARDS) patients are provided a lung rest strategy during extracorporeal membrane oxygenation (ECMO) treatment, the exact conditions of barotrauma is unclear. Therefore, we analyzed the epidemiology and risk factors for barotrauma in ARDS patients using ECMO in a single, large ECMO center in China. METHODS: A retrospective analysis was performed on 127 patients with ARDS received veno-venous (VV) ECMO who met the Berlin definition. The epidemiology and risk factors for barotrauma during ECMO were analyzed. RESULTS: Among 127 patients with ARDS treated with ECMO, barotrauma occurred in 24 (18.9%) during ECMO and 9 (7.1%) after ECMO decannulation, mainly in the late stage of ARDS (75%) and ≥8 days during ECMO (54.2%). Univariate and multivariate analyses showed that younger ARDS patients (OR = 0.953, 95%CI 0.923-0.983, p = 0.003) and those with pneumocystis jirovecii pneumonia (PJP) (OR = 3.15, 95%CI 1.070-9.271, p = 0.037), elevated body temperature after establishing ECMO (OR = 2.997, 95%CI 1.325-6.779, p = 0.008) and low platelet count after establishing ECMO (OR = 0.985, 95%CI 0.972-0.998, p = 0.02) had an increased risk of barotrauma during ECMO. There was no difference in ventilator parameters between patients with and without barotrauma. Barotrauma during ECMO was mainly related to the etiology of the disease and disease state. CONCLUSION: There is a high incidence of barotrauma in ARDS patients during ECMO, even after ECMO decannulation. Young age, PJP, elevated body temperature and low platelet count after establishing ECMO are risk factors of barotrauma, and those patients should be closely monitored by imaging, especially in the late stage of ARDS.

Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression.

The effects of rosiglitazone (RSG) in patients with type 2 diabetes mellitus (T2DM) remain controversial. Here, we first used network pharmacology to identify the common targets of RSG in the treatment of diabetes angiopathy (DA). Enrichment analysis found that the common genes were involved in the inflammatory response, leukocyte cell-cell adhesion, mitochondrion organization and oxidative stress. Our previous research confirmed that heat shock protein 22 (HSP22) suppresses diabetes-induced endothelial activation and injury by inhibiting mitochondrial reactive oxygen species (mtROS) formation and dysfunction. We then constructed HSP22 knockout mice with T2DM to investigate whether RSG protected the vascular endothelium by upregulating HSP22. Our study suggested that RSG reduced vascular endothelial cell activation and injury by decreasing monocyte adhesion and cytokine secretion and simultaneously upregulating HSP22 expression. Mechanistically, RSG inhibited mitochondrial oxidative stress and dysfunction by regulating PPAR-γ in a manner partially dependent on expression of HSP22, resulting in reduced DA.

Dynamic analysis of gene signatures in the progression of COPD.

Aims: Oxidative stress is an important amplifying mechanism in COPD; however, it is unclear how oxidative stress changes and what its exact amplification mechanism is in the pathological process. We aimed to dynamically analyse the progression of COPD and further elucidate the characteristics of each developmental stage and unveil the underlying mechanisms. Methods: We performed a holistic analysis by integrating Gene Expression Omnibus microarray datasets related to smoking, emphysema and Global Initiative for Chronic Obstructive Lung Disease (GOLD) classification based on the concept of gene, environment and time (GET). Gene ontology (GO), protein-protein interaction (PPI) networks and gene set enrichment analysis (GSEA) were used to explore the changing characteristics and potential mechanisms. Lentivirus was used to promote HIF3A overexpression. Results: In smokers versus nonsmokers, the GO term mainly enriched in "negative regulation of apoptotic process". In later transitions between stages, the main enriched terms were continuous progression of "oxidation-reduction process" and "cellular response to hydrogen peroxide". Logistic regression showed that these core differentially expressed genes (DEGs) had diagnostic accuracy in test (area under the curve (AUC)=0.828) and validation (AUC=0.750) sets. GSEA and PPI networks showed that one of the core DEGs, HIF3A, strongly interacted with the ubiquitin-mediated proteolysis pathway. Overexpression of HIF3A restored superoxide dismutase levels and alleviated the reactive oxygen species accumulation caused by cigarette smoke extract treatment. Conclusion: Oxidative stress was continuously intensified from mild emphysema to GOLD 4; thus, special attention should be paid to the identification of emphysema. Furthermore, the downregulated HIF3A may play an important role in the intensified oxidative stress in COPD.

Using machine learning for the early prediction of sepsis-associated ARDS in the ICU and identification of clinical phenotypes with differential responses to treatment.

Background: An early diagnosis model with clinical phenotype classification is key for the early identification and precise treatment of sepsis-associated acute respiratory distress syndrome (ARDS). This study aimed to: 1) build a machine learning diagnostic model for patients with sepsis-associated ARDS using easily accessible early clinical indicators, 2) conduct rapid classification of clinical phenotypes in this population, and 3) explore the differences in clinical characteristics, outcomes, and treatment responses of different phenotypes. Methods: This study is based on data from the Telehealth Intensive Care Unit (eICU) and Medical Information Mart for Intensive Care IV (MIMIC-IV). We trained and tested the early diagnostic model of sepsis-associated ARDS patients in the eICU. We used key predictive indicators to cluster sepsis-associated ARDS patients and determine the characteristics and clinical outcomes of different phenotypes, as well to explore the differences of in-hospital mortality among different the positive end-expiratory pressure (PEEP) levels in different phenotypes. These results are verified in MIMIC-IV to evaluate whether they are repeatable. Results: Among the diagnostic models constructed in 19,249 sepsis patients and 5,947 sepsis-associated ARDS patients, the AdaBoost (Decision Tree) model achieved the best performance with an area under the receiver operating characteristic curve (AUC) of 0.895, which is higher than that of the traditional Logistic Regression model (Z = -2.40,p = 0.013), and the accuracy of 70.06%, sensitivity of 78.11% and specificity of 78.74%. We simultaneously identified three sepsis-associated ARDS phenotypes. Cluster 0 (n = 3,669) had the lowest in-hospital mortality rate (6.51%) and fewer laboratory abnormalities (lower WBC (median:15.000 K/mcL), lower blood glucose (median:158.000 mg/dl), lower creatinine (median:1.200 mg/dl), lower lactic acid (median:3.000 mmol/L); p < 0.001). Cluster 1 (n = 1,554) had the highest in-hospital mortality rate (75.29%) and the most laboratory abnormalities (higher WBC (median:18.300 K/mcL), higher blood glucose (median:188.000 mg/dl), higher creatinine (median:2.300 mg/dl), higher lactic acid (median:3.900 mmol/L); p < 0.001). Cluster 2 (n = 724) had the most complex condition, with a moderate in-hospital mortality rate (29.7%) and the longest intensive care unit stay. In Clusters 0 and 1, patients with high PEEP had higher in-hospital mortality rate than those with low PEEP, but the opposite trend was seen in Cluster 2. These results were repeatable in 11,935 patients with sepsis and 2,699 patients with sepsis-associated ARDS patients in the MIMIC-IV cohort. Conclusion: A machine learning diagnostic model of sepsis-associated ARDS patients was established. Three phenotypes with different clinical features and outcomes were clustered, and these had different therapeutic responses. These findings are helpful for the early and rapid identification of sepsis-associated ARDS patients and their precise individualized treatment.

IL35 attenuated LPS-induced acute lung injury by regulating macrophage polarization.

BACKGROUND: Interleukin 35 (IL35) has been reported to play a role in acute lung injury (ALI); however, the current results regarding the relationship between IL35 and ALI are inconsistent. Therefore, we aimed to further determine the function of IL35 in ALI in mice and the potential mechanism in this paper. MATERIALS AND METHODS: Hematoxylin-eosin (HE) staining and Masson staining were used to evaluate lung injury in mice. Immunohistochemical staining was used to evaluate the expression of IL35 p35, TLR4 and MD2 and the Bax/Bcl2 and p-P65/P65 ratios. The expression levels of IL35 EBi3, CD68, CD206 and MPO were assessed by immunofluorescence staining. RT-PCR was used to examine the expression levels of IL1ß and IL6. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was performed to detect apoptotic cells. RESULTS: Overexpression of IL35 alleviated LPS-induced ALI in mice. IL35 overexpression decreased the expression of CD68 and increased the expression of CD206 in mice with ALI. Furthermore, upregulation of IL35 expression obviously reduced the expression of MPO, IL1ß and IL6 in the lung tissues of mice with ALI. Mechanistically, IL35 suppressed the TLR4/NFκB-P65 pathway, leading to the promotion of the M1 to M2 macrophage transition and alleviation of inflammation in mice with ALI. CONCLUSIONS: IL35 relieved LPS-induced inflammation and ALI in mice by regulating M1/M2 macrophage polarization and inhibiting the activation of the TLR4/NFκB-P65 pathway.

Microarray Analysis Reveals the Changes in Circular RNA Expression and Molecular Mechanisms in Mice With Ventilator-Induced Lung Injury.

Circular RNA (circRNA) expression profiles in lung tissues from mice with and without ventilator-induced lung injury (VILI) were analyzed using high-throughput sequencing and bioinformatics to clarify their potential role in VILI pathogenesis and provide valuable molecular markers for VILI diagnosis and treatment. A VILI mouse model was established using high-tidal volume ventilation, and lung tissue was stained with HE and TUNEL. The present study used high-throughput sequencing technology to analyze the expression profile of circRNAs in the lung tissue of mice with and without VILI. Bioinformatics was used to analyze the enrichment of differentially expressed circRNAs using Gene Ontology and KEGG to predict function. Among the top 10 circRNAs with significant differential expression, we used real-time quantitative polymerase chain reaction technology (qRT-PCR) to verify the accuracy of the high-throughput sequencing results and constructed the corresponding circRNA-miRNA-mRNA-specific binding network map using software prediction. The most upregulated circRNAs were novel_circ_0000899 and novel_circ_0014815, and the most downregulated circRNAs were novel_circ_0015069. A total of 14,347 circRNAs were detected using high-throughput sequencing. Compared to the control group, 285 circRNAs were abnormally and significantly expressed in the lung tissues of VILI mice (|log2(FC)| > 1, p < 0.05). A total of 171 circRNAs were significantly upregulated, and 114 circRNAs were significantly downregulated. Gene ontology analyses indicated that the differentially expressed circRNAs were involved in multiple biological functions, such as regulation of metabolic processes, protein phosphorylation, and chromatin organization. KEGG pathway analyses revealed that the Ras signaling pathway, rap1 signaling pathway, PI3K - Akt signaling pathway, and ECM receiver interaction were related to the differentially expressed circRNAs. The qRT-PCR verification results were generally consistent with the circRNA expression trends of the high-throughput sequencing data. The circRNA-miRNA-mRNA interaction network suggested that miRNAs and mRNAs related to circRNAs played a key role in VILI. Differentially expressed circRNAs were identified in the tissues of VILI mice using high-throughput sequencing combined with bioinformatics analysis, and the results lay a foundation for further study of the mechanism of circRNAs in the occurrence and development of VILI.

[Bioinformatics analysis of ventilator-induced lung injury genome microarray based on gene expression omnibus database and key gene verification].

OBJECTIVE: To investigate differential expression gene (DEG) in mice with ventilator-induced lung injury (VILI) by bioinformatics analysis, and to verify the key genes by reproducing the VILI mouse model. METHODS: (1) Experiment 1 (bioinformatics analysis): the microarray dataset of GSE9368 and GSE11662 regarding VILI mice and those in the spontaneous breathing control group were downloaded from the gene expression omnibus (GEO) database. DEG obtained by R and Venn map was further used to obtain common DEG. DAVID online database was used to obtain gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Finally, the protein-protein interaction (PPI) analysis of common DEG was carried out by using Search Tool for the Retrieval of Interacting Genes Database (STRING) and the key genes were screened out by using CytoScape software, molecular complex detection (MCODE) analysis plug-in and CytoHubba plug-in with maximum cluster centrality (MCC), maximum neighbor connectivity (MNC) and degree. (2) Experiment 2 (related protein verification): VILI mouse model was reproduced by high tidal volume (20 mL/kg) ventilator. Spontaneous breathing control group was set up. Hematoxylin-eosin (HE) staining was performed to assess lung injury and the key genes screened in experiment 1 were verified by immunohistochemical staining. RESULTS: (1) Experiment 1 results: a total of 114 DEG were screened from GSE9368 dataset, including 99 up-regulated genes and 15 down-regulated genes. A total of 258 DEG were screened from GSE11662 dataset, including 188 up-regulated genes and 70 down-regulated genes. Furthermore, 66 common DEG were obtained, including 61 up-regulated genes and 5 down-regulated genes. GO analysis showed that the common DEG were mainly involved in inflammatory response, immune response, leukocyte and neutrophil chemotaxis. KEGG analysis showed that the common DEG were involved cell adhesion, cytokine receptor interaction and tumor necrosis factor (TNF) signaling pathway. STRING and CytoScape analysis were used to construct gene PPI network diagram and important sub modules. And the CytoHubba plug-in with MCC, MNC and degree algorithms was used to perform topology analysis and then taken an intersection to obtain eight genes including suppressor of cytokine signaling 3 (SOCS3), interleukin-1ß (IL-1ß), matrix metalloproteinase-9 (MMP-9), integrin Itgam, CXC chemokine ligand 2 (CXCL2), CXC chemokine receptor 2 (CXCR2), Sell and CC chemokine receptor 1 (CCR1). (2) Experiment 2 results: a mouse model of high tidal volume VILI was reproduced. Compared with the spontaneous breathing control group, the lung tissue was injured slightly at 0 hour after the end of ventilation, and the lung tissue structure was significantly damaged at 6 hours after the end of ventilation, showing bleeding in alveolar cavity, significant increase and collapse of alveolar wall thickness, and infiltration of inflammatory cells. The top three genes from intersection and topological analysis including IL-1ß, SOCS3 and MMP-9 were verified by immunohistochemical staining. The results showed that the expressions of IL-1ß, SOCS3 and MMP-9 were gradually increased with time of ventilation, the differences were found at 6 hours as compared with those in the spontaneous breathing control group [IL-1ß (integral A value): 8.40±2.67 vs. 5.10±0.94, SOCS3 (integral A value): 9.74±1.80 vs. 5.95±1.31, MMP-9 (integral A value): 11.45±6.20 vs. 5.36±1.28, all P < 0.05]. CONCLUSIONS: Bioinformatics analysis based on GSE9368 and GSE11662 data sets found that VILI is mainly related to inflammatory injury, cytokines and immune cell infiltration; IL-1ß, SOCS3 and MMP-9 might be biomarkers of VILI.

TMT-Based proteomics analysis of LPS-induced acute lung injury.

PURPOSE: The proteome during lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice is unclear. MATERIALS AND METHODS: In this study, eight-week-old male C57BL/6 mice were intraperitoneally injected with LPS and sacrificed 18 hours after LPS administration to identify protein expression levels in lung tissue using tandem mass tag (TMT) analysis for relative quantification. Hematoxylin-eosin (HE) staining was used to evaluate lung injury in mice. Immunohistochemical staining was used to calculate the production of myeloperoxidase (MPO) and TUNEL staining was performed to detect apoptosis. GO functional clustering and KEGG pathway enrichment analyses were performed to determine functions of differentially expressed proteins (DEPs) and transduction pathways. Domain annotation and subcellular localization analysis of the DEPs were also performed. Furthermore, parallel reaction monitoring (PRM) analysis was used to verify the top 30 DEPs. RESULTS: A total of 5188 proteins were found to be expressed in lung tissues from LPS- and saline-treated mice. Among these proteins, 293 were differentially expressed between the two groups; 255 proteins were upregulated in the LPS-treated ALI mice, while 38 were downregulated. GO analysis showed that the DEPs are mainly extracellular, and KEGG analysis suggested that the DEPs are mainly enriched in the NOD-like receptor signaling pathway, complement and coagulation cascades and natural killer cell-mediated cytotoxicity. Enrichment of the DEPs is mainly peptidase S1A, serine proteases, peptidase S1, and the serpin domain. 26.6% of the DEPs are in the nucleus, 24.6% are in the cytosol, 19.1% are in the extracellular space, and 18.8% are in the plasma membrane. PRM validation showed that the trend of 30 DEPs was same with TMT analysis. Among these, Cytochrome b-245 heavy chain (Cybb), Monocyte differentiation antigen CD14 (Cd14) and Neutrophil gelatinase-associated lipocalin (NGAL) were the most obvious change. CONCLUSIONS: Our results may help to identify markers and therapeutic targets for LPS-induced ALI.

Heat shock protein B8 promotes proliferation and migration in lung adenocarcinoma A549 cells by maintaining mitochondrial function.

Heat shock protein B8 (HSPB8) impacts on tumor proliferation and migration of malignancy. However, the role of HSPB8 in lung adenocarcinoma (LUAC) remains unclear. The aim of this study, therefore, was to clarify whether HSPB8 could bring benefits to proliferation and migration of LUAC and its underlying mechanisms. The expression of HSPB8 was first evaluated by immunohistochemistry in 35 LUAC samples. Then, A549 lung adenocarcinoma cells were transfected with pcDNA-HSPB8 or si-HSPB8 to induce HSPB8 overexpression and silence. Cellular activity was evaluated with a Cell Counting Kit-8 (CCK-8) assay. Cell proliferation and migration were observed by EdU assay and scratch assay. Mitochondria-specific reactive oxygen species (mtROS) and membrane potential were measured using MitoSOX Red probe and JC-1 staining. Superoxide dismutase (SOD) activities and malondialdehyde (MDA) level were measured using commercial kits, respectively. HSPB8 protein, mitochondrial fusion protein MFN2 and mitochondrial fission protein p-Drp1/Drp1 were measured using western blot. Compared with the normal tissues, the expression of HSPB8 protein was higher in LUAC tissues and upregulation of HSPB8 protein was related to tumor size and tumor location. Furthermore, HSPB8 overexpression aggravated cell proliferation and migration of A549 cells. Mechanistically, HSPB8 suppressed mitochondrial impairment, leading to promoting the progress of A549 lung adenocarcinoma cells. These data demonstrate that HSPB8 plays an important role in progression of LUAC and may be a new target to treat LUAC.

Maternal embryonic leucine zipper kinase: A novel biomarker and a potential therapeutic target in lung adenocarcinoma.

Maternal embryonic leucine zipper kinase (MELK), is an adenosine monophosphate-activated protein kinase-related kinase that serves important roles in tumourigenesis in multiple malignant tumours. However, to the best of our knowledge, the effect of MELK in lung adenocarcinoma (LUAD) has not been elucidated. The present study aimed to explore the clinical significance of MELK in the prognosis of LUAD. Data from Oncomine, Gene Expression Profiling Interactive Analysis (GEPIA) and The Cancer Genome Atlas (TCGA) were selected to predict the differential mRNA expression levels of MELK mRNA in LUAD and normal tissues. Subsequently, LUAD and adjacent normal tissue samples were collected from 75 patients with the disease, and immunohistochemistry was used to detect the protein expression of MELK. In addition, the Kaplan-Meier Plotter database, GEPIA and TCGA were used to verify the effect of MELK expression on clinical prognosis in patients with LUAD. MELK was significantly upregulated in LUAD tissues compared with that in normal tissues based on Oncomine, GEPIA and TCGA data (P<0.05). In addition, the results from immunohistochemistry demonstrated that the MELK protein level in LUAD tissues was significantly higher compared with that in matched normal tissues (P<0.05). Prognostic analysis performed using the Kaplan-Meier plotter, GEPIA and TCGA suggested that the expression of MELK was negatively associated with the overall survival time of patients with LUAD (P<0.05). In conclusion, MELK was highly expressed in LUAD based on bioinformatics and immunohistochemistry analysis, and increased expression of MELK was associated with a poor patient prognosis. MELK may serve as a potential diagnostic marker and therapeutic target for LUAD.

The expression and prognostic significance of Drp1 in lung cancer: A bioinformatics analysis and immunohistochemistry.

BACKGROUND: Dynamin-related protein 1 (Drp1) plays important roles in tumorigenesis, including lung cancer. However, the effect of Drp1 in lung cancer remains unclear. The present study was aimed to investigate the clinical significance and effect of Drp1 on prognosis of lung cancer. METHODS: Oncomine and The Cancer Genome Atlas (TCGA) databases were selected to predict the differential expression levels of Drp1 in lung cancer. Then, 70 cases of lung cancer and normal tissues were collected and immunohistochemistry was used to detect the expression of Drp1. In addition, Kaplan-Meier Plotter database and TCGA database were used to verify the correlation between Drp1 expression and the clinical prognosis in lung cancer patients. RESULTS: Drp1 was significantly overexpressed in lung cancer tissues based on Oncomine and TCGA databases (P < .05). Moreover, results from immunohistochemistry showed that Drp1 protein level in lung cancer was also significantly higher than that in the matched normal tissues (P < .05). Prognostic analysis from Kaplan-Meier Plotter database with the chosen probe IDs of 203105_s_at suggested that Drp1 was negatively correlated to overall survival (OS) of lung cancer patients (HR = 1.16, 95% CI: 1.02-1.31; P = .025), but not in the probe IDs of 226154_at (HR = 0.86, 95% CI: 0.73-1.01; P = .069). However, prognosis from TCGA database showed inconsistent results in which high expression of Drp1 was correlated with worse survival probability of all, male, female in lung adenocarcinoma (P < .05), but not in LUSC (P > .05). CONCLUSION: Drp1 was highly expressed in lung cancer based on bioinformatics analysis and tissue microarray, but there was a lot of inconsistency in prognosis depending on different levels of Drp1 from the bioinformatics analysis.

Peptide nucleic acids targeting mitochondria enhances sensitivity of lung cancer cells to chemotherapy.

Acquired resistance to chemotherapy is a major limitation for the successful treatment of lung cancer. Previously, we and others showed that formation of tumor spheres is associated with chemotherapy resistance in lung cancer cells, but the underlying mechanisms remained largely unknown. In the current study, we show that mitochondrial activity is significantly higher in A549 tumor spheres versus monolayer cells, establishing mitochondria as a putative target for antitumor therapy. To this end, we designed a peptide nucleic acids (PNAs) coupled with triphenylphosphonium (TPP) to target the displacement loop (D-loop) regulatory region of mitochondrial DNA (PNA-mito). Treatment with PNA-mito significantly disrupted mitochondrial gene expression, inhibited membrane potential and mitochondria fusion, resulting in proliferation inhibition and cell death. Consistently, in mouse xenograft models, PNA-mito could efficiently inhibit mitochondrial gene expression and block tumor growth. Treatment with a low dose of PNA-mito could significantly enhance the chemotoxicity of cisplatin (CDDP) in drug-resistant A549 tumor spheres. These results establish mitochondria-targeting PNAs as a novel strategy to enhance the accumulative therapeutic outcome of lung cancer.

The role of lncRNA CASC2 on prognosis of malignant tumors: a meta-analysis and bioinformatics.

Background: Cancer susceptibility candidate 2 (CASC2) is characterized as a tumor suppressor, which was first identified to be downregulated in endometrial carcinoma. Accumulating evidence was provided to testify the function of CASC2 in malignant tumors. However, a systematic and quantitative assessment is not available. The present study was designed to evaluate the role of CASC2 in multiple carcinomas through meta-analysis and bioinformatics. Materials and methods: A systematic assessment of the relationship of CASC2 with tumors was performed by using several computerized databases from inception to December 1, 2017. Pooled HR with 95% CI was calculated to summarize the effect. The data on prognosis of malignant tumors were also downloaded from The Cancer Genome Atlas (TCGA) project, OncoLnc, TANRIC and lncRNAtor database. Results: A total of 13 studies with 966 cancer patients were pooled in the analysis to evaluate the prognostic value of CASC2 in multiple tumors and the clinical features. The results of the meta-analysis revealed that low expression levels of CASC2 were associated with poor overall survival (OS) (pooled HR=0.39, 95% CI: 0.28-0.53, P<0.0001). CASC2 obviously has a negative correlation with advanced tumor node metastasis (TNM) stage, lymph node metastasis (LNM) and T stage, respectively (P<0.05). There was, however, no significant difference in gender, distant metastasis and high differentiation (P>0.05). In the Kaplan-Meier curves with log-rank analysis, higher expression of CASC2 was positively correlated with longer survival time than patients with a lower level (P<0.05), including kidney renal clear cell carcinoma, brain lower grade glioma, pancreatic adenocarcinoma and sarcoma. Conclusion: Findings from this meta-analysis suggest that lower expression of CASC2 is associated with poorer prognosis of cancers, as well as advanced TNM, LNM and T stage. Data from the bioinformatics analysis revealed that higher expression of CASC2 was related to longer OS in patients with malignant tumors.

A case of herbicide-induced acute fibrinous and organizing pneumonia?

BACKGROUND: To improve the understanding of acute fibrinous and organizing pneumonia (AFOP), we present one case of AFOP proven by percutaneous lung biopsy along with clinical features, chest imaging and pathology. CASE PRESENTATION: A 50-year-old man was admitted to our department after he was given empiric therapy for community-acquired pneumonia (CAP). The clinical symptoms of the patient were dry cough, chills, night sweats and high fevers. Chest computed tomography (CT) scan showed a high-density shadow in the right lung lobe, similar to lobular pneumonia. The patient was preliminarily diagnosed with community-acquired pneumonia; however, antibacterial treatment was ineffective. To confirm the diagnosis, we performed bronchoscopy and percutaneous lung biopsy; pathology was consistent with AFOP. After he was treated with glucocorticoids, the patient's symptoms were relieved, and the shadow seen on imaging dissipated during the follow-up period. CONCLUSIONS: AFOP is a rare histopathological diagnosis that can be easily misdiagnosed. Clinicians need to consider the possibility of AFOP in the case of invalid antibacterial therapy.

Endobronchial hamartoma mimicking malignant lung tumor contralateral endobronchial metastasis: A case report.

RATIONALE: Endobronchial hamartoma, the most common benign lung tumor, is located in the bronchus, and it easily mimics lung cancer or bronchial metastasis. Endobronchial hamartoma can cause coughing, hemoptysis, and pulmonary infection; thus, it should be treated right away by surgery or fiberoptic bronchoscopy. PATIENT CONCERNS: We report a rare case of endobronchial hamartoma in which the clinical symptoms and imaging overlapped strongly with malignant lung tumor contralateral endobronchial metastasis. DIAGNOSES: Endobronchial hamartoma coexisting with a malignant lung tumor. INTERVENTIONS: Fiberoptic bronchoscopy was conducted, and the pathologic diagnosis was hamartoma. A second fiberoptic bronchoscopy was conducted, and fine-needle aspiration cytology of the enlarged lymph nodes indicated squamous cell carcinoma. OUTCOMES: The clinical symptoms were relieved, and the treatment options were docetaxel, cis-dichlorodiamineplatinum, and endostatin. LESSONS: Fiberoptic bronchoscopy needs to be guided by imaging and can be considered an effective method for the diagnosis of endobronchial hamartoma.

Rapamycin inhibited the function of lung CSCs via SOX2.

The presence of cancer stem cells (CSCs) is the source of occurrence, aggravation, and recurrence of lung cancer. Accordingly, targeting killing the lung CSCs has been suggested to be an effective approach for lung cancer treatment. In this study, we showed that rapamycin inhibited the mammalian target of rapamycin (mTOR) signal transduction in A549 cells and improved the sensitivity to cisplatin (DDP). The mechanisms involve inhibition of the SOX2 expression, cell proliferation, epithelial-mesenchymal transition (EMT) phenotype, and sphere formation. Interestingly, knocked down SOX2 was a similar effect with rapamycin in A549 sphere. Furthermore, we showed that ectopic expression of Sox2 in A549 cells was sufficient to render them more resistant to rapamycin treatment in vitro. These data suggested that rapamycin inhibited the function of lung CSCs via SOX2. It will be of great interest to further explore the therapeutic strategies of lung cancer.

The association between MMP-12 82 A/G polymorphism and susceptibility to various malignant tumors: a meta-analysis.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases responsible for degrading essentially all components of the extracellular matrix (ECM). Accumulating evidence suggests that MMPs might play a critical role in growth, invasion, and metastasis of malignant tumors. A single nucleotide polymorphism (SNP) in the promoter region of MMP-12, MMP-12 82 A/G (rs2276109), has been recognized to play a critical role in regulating the expression of MMP-12, however, its correlation with tumor susceptibility remains controversial. To address this issue, we performed meta-analysis to investigate the association MMP-12 82 A/G polymorphism and susceptibility of nine malignant tumors from 11 studies, including 6153 cancer patients and 6838 controls. Two reviewers independently screened studies for eligibility and extracted data for included studies. While overall no evident association between MMP-12 82 A/G and tumor susceptibility was observed, subgroup analysis revealed a specific role of G allele in increasing the susceptibility for epithelial ovarian carcinoma (EOC) using the allele model (fixed effects OR = 2.45, 95% CI = 1.46-4.10, P = 0.001) and the dominant model (fixed effects OR = 2.52, 95% CI = 1.49-4.24, P = 0.001). We thus suggest that G allele of MMP-12 82 A/G polymorphism is a genetic risk factor for EOC.