Real-time, random-access organ screening for carbapenem-resistant organisms (CRO) reduces CRO-associated, donor-derived infection mortality in lung transplant recipients.

PURPOSE: Donor-derived infection (DDI) has become an important factor affecting the prognosis of lung transplantation patients. The risks versus benefits of using donor organs infected with multidrug-resistant organisms (MDRO), especially carbapenem-resistant organisms (CRO), are frequently debated. Traditional microbial culture and antimicrobial susceptibility testing at present fail to meet the needs of quick CRO determination for donor lungs before acquisition. In this study, we explored a novel screening method by using Xpert® Carba-R assay for CRO in donor lungs in a real-time manner to reduce CRO-associated DDI mortality. METHODS: This study was registered on chictr.org.cn (ChiCTR2100053687) on November 2021. In the Xpert Carba-R screening group, donor lungs were screened for CRO infection by the Xpert Carba-R test on bronchoalveolar fluid (BALF) before acquisition. If the result was negative, donor lung acquisition and subsequent lung transplantation were performed. In the thirty-five potential donors, nine (25.71%) with positive Xpert Carba-R results in BALF were declined for lung transplantation. Twenty-six recipients and the matching CRO-negative donor lungs (74.29%) were included in the Xpert Carba-R screening group. In the control group, nineteen recipients underwent lung transplants without Xpert Carba-R screening. The incidence and mortality of CRO-associated DDI were collected and contrasted between the two groups. RESULTS: Multivariate analysis showed that CRO-related death due to DDI within 60 days was significantly lower in the Xpert Carba-R screening group than that in the control group (OR = 0.05, 95% CI 0.003-0.74, p = 0.03). CONCLUSION: Real-time CRO screening of donor lungs before transplantation at the point of care by the Xpert Carba-R helps clinicians formulate lung transplantation strategies quickly and reduces the risk of subsequent CRO infection improving the prognosis of lung transplantation.

Multiplex digital PCR with digital melting curve analysis on a self-partitioning SlipChip.

Digital polymerase chain reaction (digital PCR) can provide absolute quantification of target nucleic acids with high sensitivity, excellent precision, and superior resolution. Digital PCR has broad applications in both life science research and clinical molecular diagnostics. However, limited by current fluorescence imaging methods, parallel quantification of multiple target molecules in a single digital PCR remains challenging. Here, we present a multiplex digital PCR method using digital melting curve analysis (digital MCA) with a SlipChip microfluidic system. The self-partitioning SlipChip (sp-SlipChip) can generate an array of nanoliter microdroplets with trackable physical positions using a simple loading-and-slipping operation. A fluorescence imaging adaptor and an in situ thermal cycler can be used to perform digital PCR and digital MCA on the sp-SlipChip. The unique signature melting temperature (Tm) designed for amplification products can be used as a fingerprint to further classify the positive amplification partitions into different subgroups. Amplicons with Tm differences as low as 1.5 degrees celsius were clearly separated, and multiple amplicons in the same partition could also be distinguished by digital MCA. We further demonstrated this digital MCA method with simultaneous digital quantification of five common respiratory pathogens, including Staphylococcus aureus, Acinetobacter baumannii, Streptococcus pneumoniae, Hemophilus influenzae, and Klebsiella pneumoniae. Since digital MCA only requires an intercalation dye instead of sequence-specific hydrolysis probes to perform multiplex digital PCR analysis, it can be less expensive and not limited to the number of fluorescence channels.

Gene regulation and prognostic indicators of lung squamous cell carcinoma: TCGA-derived miRNA/mRNA sequencing and DNA methylation data.

Lung squamous cell carcinoma (LSCC) is a common cancer worldwide, and this study aimed to investigate the key regulatory networks and prognostic indicators of LSCC. MicroRNA (miRNA)/messenger RNA (mRNA) sequencing and DNA methylation data were obtained from the Cancer Genome Atlas. Differentially expressed miRNAs (DEmiRNAs) and genes (DEGs) were identified by the limma package. Then, the transcription factors (TFs) of DEmiRNAs/DEGs, as well as the targets of miRNAs, were predicted by the TFmiR online tool. Using the t test, aberrant methylation was detected in TF binding sites (TFBSs) in promoters. Finally, integrated network and survival analyses were conducted using SPSS software. We obtained 104 DEmiRNAs and 4,491 DEGs, and validated 2,113 DEGs (VDEGs). Then, 103 TFs, 295 TFs, and 14 DEmiRNAs were predicted to target 95 DEmiRNAs, 821 DEGs and 283 DEGs, respectively. After TF-DEmiRNA/DEG and TF-DEmiRNA-DEG networks were constructed (e.g., E2F1-CDC25A, miR29a-RAN, miR326-TBL1XR1), five feedforward loops between ZEB1 and miR-141/200a/200b/200c/429 were found. Furthermore, VDEGs CDC25A, RAN, TBL1XR1 as well as miR-130b and miR-590 were negatively correlated with survival rates. E2F1-CDC25A, miR29a-RAN, miR326-TBL1XR1, and the feedforward loops between ZEB1/ZEB2 and miR-141/200a/200b/200c/429 might participate in LSCC development. Compared with BEAS-2B cells, the SK-MES-1 cells presented a higher expression level of miR-141, miR-200a, miR-200b, miR-200c but a lower expression level of ZEB1. Overexpressed miR-200c significantly attenuated the expression of ZEB1 and ZEB2 and inhibited the proliferation and migration of SK-MES-1 cells (all p < 0.05). In addition, CDC25A, miR-200a, miR-200b, miR-200c, miR-130b, and miR-590 are potential prognostic indicators of LSCC.

Cavity Formation is a Prognostic Indicator for Pathologic Stage I Invasive Lung Adenocarcinoma of ≥3 cm in Size.

BACKGROUND We investigated the correlation between cavity formation, prognosis, and tumor stage for pathologic stage I invasive lung adenocarcinomas (IADCs) ≤3 cm in size. MATERIAL AND METHODS 2106 candidates with pathologic stage I IADC were identified from Shanghai Chest Hospital between 2009 and 2014. There were 227 patients who were diagnosed as having cavity formation and another 1879 patients who were not (the non-cavitary lung cancer group). Kaplan-Meier analysis curves were conducted to compare the overall survival (OS) and relapse-free survival (RFS) between these 2 groups. Cox proportional hazards regression was performed to discover the independent risk factors of OS and RFS. Receiver operating characteristic (ROC) curve was done to determine the cutoff value of cavity size for predicting prognosis. Furthermore, subgroup analysis was stratified by the size of tumor and the 8th classification of T category. RESULTS Compared with non-cavitary lung cancer group, patients with cavity formation were found to have a higher prevalence of male patients (P=0.015), older age patients (P=0.039), larger size tumors (P=0.004), and worse cancer relapse (P<0.001). Survival analysis found that patients with cavitary IADC had significantly shorter RFS than those with non-cavitary IADC (P=0.001). Further, subgroup analysis confirmed a significantly worse RFS in cavitary IADC group both in stage T1a (P=0.002) and T1b (P<0.001), but not for stage T1c (P=0.962) and T2a (P=0.364). Moreover, cavity formation was still less of a significant predictor of RFS in multivariable analysis (hazard ratio [HR] 1.810, 95% confidence level [CI] 1.229-2.665, P=0.003). The ROC curve showed that the best cutoff value of maximum diameter of the cavity for judging RFS was 5 mm (sensitivity: 0.500; specificity: 0.783). At the same time, multiple cavities were more likely to lead to recurrence (sensitivity: 0.605; specificity: 0.439). CONCLUSIONS Cavitary adenocarcinoma was a worse prognostic indicator compared with non-cavitary adenocarcinoma, especially for cavity >5 mm and multiple cavities. Thus, for stage T1a and T1b, cavitary and non-cavitary IADC should be considered separately.

Sialyltransferase ST3GAL6 mediates the effect of microRNA-26a on cell growth, migration, and invasion in hepatocellular carcinoma through the protein kinase B/mammalian target of rapamycin pathway.

Aberrant sialylation profiles on the cell surface have been recognized for their potential diagnostic value in identifying the regulation of tumor properties in several cancers, including hepatocellular carcinoma (HCC). Recently, increasing evidence has suggested that the deregulation of microRNA (miRNA) is a common feature in human cancers. In this study, we found obvious upregulation of sialyltransferase ST3GAL6 both in HCC cell lines and in tissue samples. The altered expression of ST3GAL6 was found to correlate with cell proliferation, migration, and invasion ability in HCC. Further investigation showed that miR-26a negatively regulated ST3GAL6, inducing the suppression of cell proliferation, migration, and invasion in vitro. Moreover, we identified the protein kinase B/mammalian target of rapamycin (Akt/mTOR) pathway as the target of ST3GAL6 based on Western blot analysis. Analysis of a xenograft mouse model showed that miR-26a significantly reduced tumor growth by suppressing activation of the Akt/mTOR pathway by directly targeting ST3GAL6. In conclusion, these data indicate that ST3GAL6 promotes cell growth, migration, and invasion and mediates the effect of miR-26a through the Akt/mTOR signaling pathway in HCC.

The histone demethylase PHF8 is an oncogenic protein in human non-small cell lung cancer.

PHF8 is a JmjC domain-containing protein and erases repressive histone marks including H4K20me1 and H3K9me1/2. It binds to H3K4me3, an active histone mark usually located at transcription start sites (TSSs), through its plant homeo-domain, and is thus recruited and enriched in gene promoters. PHF8 is involved in the development of several types of cancer, including leukemia, prostate cancer, and esophageal squamous cell carcinoma. Herein we report that PHF8 is an oncogenic protein in human non-small cell lung cancer (NSCLC). PHF8 is up-regulated in human NSCLC tissues, and high PHF8 expression predicts poor survival. Our in vitro and in vivo evidence demonstrate that PHF8 regulates lung cancer cell proliferation and cellular transformation. We found that PHF8 knockdown induces DNA damage and apoptosis in lung cancer cells. PHF8 promotes miR-21 expression in human lung cancer, and miR-21 knockdown blocks the effects of PHF8 on proliferation and apoptosis of lung cancer cells. In summary, PHF8 promotes lung cancer cell growth and survival by regulating miR-21.

Analysis and identification of mRNAsi­related expression signatures via RNA sequencing in lung cancer.

High stemness index scores are associated with poor survival in patients with lung cancer. Studies on the mRNA expression-based stemness index (mRNAsi) are typically conducted using tumor tissues; however, mRNAsi-related expression signatures based on cell-free RNA (cfRNA) are yet to be comprehensively investigated. The present study aimed to elucidate the gene expression profiles of tumor stemness in lung cancer tissues and corresponding cfRNAs in blood, and to assess their links with immune infiltration. Tumor tissue, paracancerous tissue, peripheral blood and lymph node samples were collected from patients with stage I-III non-small cell lung cancer and RNA sequencing was performed. The TCGAbiolinks package was used to calculate the mRNAsi for each of these four types of sample. Weighted gene co-expression network analysis and differentially expressed gene analyses were performed to investigate mRNAsi-related genes, and pathway enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology-based annotation system. In addition, the STAR-Fusion tool was used to detect fusion variants, and CIBERSORT was used to analyze the correlations of stemness signatures in tissues and blood with immune cell infiltration. The mRNAsi values in peripheral blood and lymph nodes were found to be higher than those in cancer tissues. 'Hematopoietic cell lineage' was the only KEGG pathway enriched in mRNAsi-related genes in both lung cancer tissues and peripheral blood. In addition, the protein tyrosine phosphatase receptor type C associated protein gene was the only gene commonly associated with the mRNAsi in these two types of sample. The expression of mRNAsi-related genes was increased in the dendritic and Treg cells in tumor tissues, but was elevated in Treg and CD8 cells in the blood. In conclusion, cfRNAs in the blood exhibit unique stemness signatures that have potential for use in the diagnosis of lung cancer.

Identification of candidate genes for lung cancer somatic mutation test kits.

Over the past three decades, mortality from lung cancer has sharply and continuously increased in China, ascending to the first cause of death among all types of cancer. The ability to identify the actual sequence of gene mutations may help doctors determine which mutations lead to precancerous lesions and which produce invasive carcinomas, especially using next-generation sequencing (NGS) technology. In this study, we analyzed the latest lung cancer data in the COSMIC database, in order to find genomic "hotspots" that are frequently mutated in human lung cancer genomes. The results revealed that the most frequently mutated lung cancer genes are EGFR, KRAS and TP53. In recent years, EGFR and KRAS lung cancer test kits have been utilized for detecting lung cancer patients, but they presented many disadvantages, as they proved to be of low sensitivity, labor-intensive and time-consuming. In this study, we constructed a more complete catalogue of lung cancer mutation events including 145 mutated genes. With the genes of this list it may be feasible to develop a NGS kit for lung cancer mutation detection.

[Correlation analysis between imaging features and lymph node metastasis in T1a lung adenocarcinoma].

OBJECTIVE: To analyze the data of patients with clinical stage T1a lung adenocarcinoma and find the predictive factors associated with lymph node metastasis. METHODS: From January to June 2012, 271 patients with small nodules of peripheral lung adenocarcinoma were enrolled in the retrospective review. There were 105 male and 112 female patients, with an average age of (61 ± 11)years (range 32-85 years). The data were collected including age, gender, smoking history, carcinoembryonic antigen(CEA), imaging findings, surgical procedure, pleural involvement, symptoms, tumor size, pathological classification, pathologic stage, maximum standardized uptake value(SUVmax) and lymph node metastasis. The predictive factors of lymph node metastasis in clinical factors were detected by univariate and multivariate analysis. RESULTS: By preoperative thin-section CT, 35 patients were categorized as pure ground-grass opacity(GGO), 11 cases of atypical adenomatous hyperplasia, 24 cases of adenocarcinoma in situ, with no lymph node metastasis. Categorized as mixed ground-glass opacities in 89 patients, 84 patients (94.4%) had no lymph node metastasis, only 5 patients (6.0%) with lymph node metastasis. Categorized as solid nodules in 93 patients, a total of 28 cases (30.1%) had lymph node metastasis. There were statistically significant difference between three groups (χ(2) = 23.41, P < 0.001) . By univariate analysis, we found that the predictive factors of lymph node metastasis were as follows: tumor size > 1 cm (χ(2) = 9.021, P < 0.003) , imaging performance with mixed GGO or solid nodules (χ(2) = 23.41, P < 0.000) , CEA > 5 µg/L (χ(2) = 15.541, P < 0.000) and PET-CT SUVmax > 5 (χ(2) = 0.644, P < 0.000). By multivariate analysis, we found that imaging performance (mixed GGO or solid nodules) was the independent predictor of lymph node metastasis in clinical factors (OR = 166.116, 95%CI:18.161-25.19, P < 0.001) . CONCLUSIONS: Patients of pure GGO generally do not have lymph node metastasis. Tumor diameter > 1 cm, imaging findings with the mixed GGO or solid nodules, carcinoembryonic antigen CEA > 5 µg/L, PET-CT SUVmax > 5 are predictive factors of lymph node metastasis in which imaging is independent predictor.

Trichosanthin-derived peptide Tk-PQ attenuates immune rejection in mouse tracheal allotransplant model by suppressing PI3K-Akt and inducing type II immune polarization.

Obliterative bronchiolitis (OB) is one of the main complications affecting long-term survival of post-lung transplantation patients. In this study, we evaluated the efficacy of Tk-PQ (a peptide derived from trichosanthin) in alleviating OB in a mouse ectopic tracheal transplant model. We found that post-transplantation treatment of Tk-PQ significant ameliorated OB symptoms including luminal occlusion, epithelial cells loss and fibrosis in the allograft. In addition, Tk-PQ promoted immune suppressive environment by inducing Th2 polarization and increasing Treg population which in turn led to elevated levels of anti-inflammatory cytokines IL-4, IL-10, IL-33 and decreased levels of pro-inflammatory IL-1ß. Mechanistically, we used transcriptome analysis of splenic T cells from allografted mice to show that Tk-PQ treatment down-regulated the PI3K-Akt signaling pathway. Indeed, the immune suppression phenotypes of Tk-PQ was recapitulated by a PI3K inhibitor LY294002. Taken together, Tk-PQ regulates post-transplantation immuno-rejection by modulating the balance of T cell response via the PI3K-Akt pathway, making it a promising peptide based immune rejection suppressant for patients receiving allotransplant.

Hydrogen saline is protective for acute lung ischaemia/reperfusion injuries in rats.

BACKGROUND: Protective effects of saturated hydrogen (H(2)) saline on cardiac ischaemia-reperfusion (I/R) injury have been demonstrated previously. This study was designed to show that hydrogen-rich saline is protective in preventing lung I/R injury in rats. METHODS: Adult male Sprague-Dawley rats underwent 45 min occlusion of the right lung roots and 120 min reperfusion. Rats were divided randomly into three groups: sham-operated control group, I/R plus saline treatment, and I/R plus hydrogen-rich saline treatment (0.6 mmol/L, 0.5 ml/kg/d). Three days of intraperitoneal injection of hydrogen-rich saline before the reperfusion combined with immediate administration of hydrogen-rich saline after the reperfusion were performed. Following reperfusion, the lung tissue and the pulmonary artery was immediately obtained and the W/D ratio, pulmonary artery contraction and relaxation ability, H-E staining, TUNEL staining, caspase-3, MDA, 8-OHdG content and measurement of such biomarkers as WBC, CRP were measured or carried out. RESULTS: Hydrogen saline significantly protected vasoactivity of the pulmonary artery, reduced pulmonary oedema, decreased lung malondialdehyde (MDA), 8-OHdG concentration, alleviated lung epithelial cell apoptosis and lowered the level of such biomarkers as WBC, CRP, ALT and TBiL. CONCLUSIONS: It is concluded that hydrogen-rich saline is a novel, simple, safe and effective method to attenuate pulmonary I/R injury.

TRIM46 activates AKT/HK2 signaling by modifying PHLPP2 ubiquitylation to promote glycolysis and chemoresistance of lung cancer cells.

The incidence of lung cancer is increasing worldwide. Although great progress in lung cancer treatment has been made, the clinical outcome is still unsatisfactory. Tripartite motif (TRIM)-containing proteins has been shown to be closely related to tumor progression. However, the function of TRIM46 in lung cancer is largely unknown. Here, TRIM46 amplification was found in lung adenocarcinoma (LUAD) tissues and TRIM46 amplification was significantly associated with a poor survival rate. Overexpression of wild type TRIM46 increased the proliferation of LUAD cells and glycolysis, promoted xenografts growth, and enhanced cisplatin (DDP) resistance of LUAD cells via increased ubiquitination of pleckstrin homology domain leucine-rich repeat protein phosphatase 2 (PHLPP2) and upregulation of p-AKT. In contrast, overexpression of RING-mutant TRIM46 did not show any effects, suggesting the function of TRIM46 was dependent on the E3 ligase activity. Furthermore, we found that TRIM46 promoted LUAD cell proliferation and DDP resistance by enhancing glycolysis. PHLPP2 overexpression reversed the effects of TRIM46 overexpression. Amplification of TRIM46 also promoted LUAD growth and enhanced its DDP resistance in a patient-derived xenograft (PDX) model. In conclusion, our data highlight the importance of TRIM46/PHLPP2/AKT signaling in lung cancer and provide new insights into therapeutic strategies for lung cancer.

Integrative Serum Metabolic Fingerprints Based Multi-Modal Platforms for Lung Adenocarcinoma Early Detection and Pulmonary Nodule Classification.

Identification of novel non-invasive biomarkers is critical for the early diagnosis of lung adenocarcinoma (LUAD), especially for the accurate classification of pulmonary nodule. Here, a multiplexed assay is developed on an optimized nanoparticle-based laser desorption/ionization mass spectrometry platform for the sensitive and selective detection of serum metabolic fingerprints (SMFs). Integrative SMFs based multi-modal platforms are constructed for the early detection of LUAD and the classification of pulmonary nodule. The dual modal model, metabolic fingerprints with protein tumor marker neural network (MP-NN), integrating SMFs with protein tumor marker carcinoembryonic antigen (CEA) via deep learning, shows superior performance compared with the single modal model Met-NN (p < 0.001). Based on MP-NN, the tri modal model MPI-RF integrating SMFs, tumor marker CEA, and image features via random forest demonstrates significantly higher performance than the clinical models (Mayo Clinic and Veterans Affairs) and the image artificial intelligence in pulmonary nodule classification (p < 0.001). The developed platforms would be promising tools for LUAD screening and pulmonary nodule management, paving the conceptual and practical foundation for the clinical application of omics tools.

Comprehensive Analysis of the Prognostic Role and Mutational Characteristics of m6A-Related Genes in Lung Squamous Cell Carcinoma.

BACKGROUND: There have been limited treatment therapies for lung squamous cell carcinoma (LUSC). M6A-related genes may be the next therapeutic targets for LUSC. In this study, we explored the prognostic role and mutational characteristics of m6A-related genes in LUSC. METHODS: LUSC gene expression data, mutational data, and corresponding clinical information were extracted from The Cancer Genome Atlas database. Differentially expressed genes (DEGs) were identified, and the mutation characteristics of LUSC patients were explored. Then, m6A-related genes were extracted and the correlations among the genes were detected. Finally, the prognostic roles of the genes were investigated and the nomogram model was developed. Besides, the protein-protein interaction (PPI) network was used to explore the potential interactions among the genes. RESULTS: In total, there are 551 LUSC samples enrolled in our study, containing 502 LUSC tumor samples and 49 adjacent normal LUSC samples, respectively. There were 2970 upregulated DEGs and 1806 downregulated DEGs were further explored. IGF2BP1 and RBM15 had significant co-occurrence frequency (p < 0.05). Besides, METTL14 and ZC3H13 or YTHDF3 also had significant co-occurrence frequency (p < 0.05). All the m6A-related genes represent the positive correlation. WTAP was identified as a prognostic gene in the TCGA database while YTHDC1 and YTHDF1 were identified as prognostic genes. In multivariate Cox analysis, YTHDF1, age, pN stage, pTNM stage, and smoking were all identified as significant prognostic factors for OS. CONCLUSION: We investigated the expression patterns and mutational characteristics of LUSC patients and identified three potential independent prognostic m6A-related genes (WTAP, YTHDC1, and YTHDF1) for OS in LUSC patients.

Prognostic impact of vascular endothelial growth factor-A and E-cadherin expression in completely resected pathologic stage I non-small cell lung cancer.

OBJECTIVE: The purpose of this study was to evaluate the value of vascular endothelial growth factor-A and E-cadherin expression as well as other confirmed prognostic factors in predicting the clinical outcome after definitive surgery of pathologic stage I non-small cell lung cancer. METHODS: One hundred and eighty-five consecutive and non-selected patients who underwent definitive surgery for stage I non-small cell lung cancer in our institute were included in this study. Formalin-fixed paraffin-embedded specimens were stained for vascular endothelial growth factor-A and E-cadherin and the correlation between the staining, its clinicopathological parameters and its prognostic power were analyzed statistically. RESULTS: Of the 185 patients studied, 92 cases (49.7%) were strongly positive for vascular endothelial growth factor-A. Vascular endothelial growth factor-A expression was only related to visceral pleural involvement (P < 0.001). A total of 95 carcinomas (51.4%) were E-cadherin-negative tumors. E-cadherin expression correlated with histology (P < 0.001), tumor size (P = 0.001) and visceral pleural involvement (P < 0.001). In univariate analysis by log-rank test, gender, tumor size, lymphovascular invasion, visceral pleural involvement, vascular endothelial growth factor-A expression and E-cadherin expression were significant prognostic factors (P = 0.003, 0.042, 0.026, 0.035, 0.008 and 0.006, respectively). In multivariate analysis, gender, vascular endothelial growth factor-A and E-cadherin expression maintained its independent prognostic influence on overall survival (P = 0.013, <0.001 and 0.036, respectively). CONCLUSIONS: Expression of vascular endothelial growth factor-A is related to visceral pleural involvement, and E-cadherin expression correlates with histology, tumor size and visceral pleural involvement. Multivariate analysis confirmed gender, vascular endothelial growth factor-A and E-cadherin expression were significant predictive factors for overall survival in completely resected pathologic stage I non-small cell lung cancer.

Progress of Clinical Application for Ex Vivo Lung Perfusion (EVLP) in Lung Transplantation.

In recent years, medical advances make lung transplantation become a standard treatment for terminal lung diseases (such as emphysema, pulmonary fibrosis, pulmonary cystic fibrosis, and pulmonary arterial hypertension) that cannot be cured by drugs or surgery (Lund et al., J Heart Lung Transplant 34:1244, 2015). However, the current number of donor lungs that meet the transplant criteria is no longer sufficient for transplanting, causing some patients to die while waiting for a suitable lung. Current methods for improving the situation of shortage of lung transplant donors include the use of donation after cardiac death (DCD) donors, smoker donors, and Ex Vivo Lung Perfusion (EVLP). Among them, EVLP is a technique for extending lung preservation time and repairing lung injury in the field of lung transplantation. By continuously assessing and improving the function of marginal donor lungs, EVLP increases the number of lungs that meet the transplant criteria and, to some extent, alleviates the current situation of shortage of donor lungs. This chapter reviews the clinical application and research progress of EVLP in the field of lung transplantation.

A comprehensive study of construction and analysis of competitive endogenous RNA networks in lung adenocarcinoma.

BACKGROUND: Long noncoding RNAs (lncRNAs) have gain increasing attention in lung adenocarcinoma. In this study, we aimed at constructing and analyzing the lncRNAs and the related proteins based competitive endogenous RNA (ceRNA) network. METHODS: RNA expression data of lung adenocarcinoma were extracted from the TCGA database. Differentially expressed (DE) lncRNAs, messenger RNAs (mRNAs) and microRNAs (miRNAs) were identified and then a DElncRNA-DEmiRNA-DEmRNA ceRNA network was constructed for lung adenocarcinoma. We also analyzed the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of the DEgenes. Kaplan-Meier survival curves were also been further utilized for exploring the prognostic factors. RESULTS: After compared and calculated lncRNA, mRNA and miRNA expression profiles between lung adenocarcinoma and normal samples, 1709 differential expressed lncRNAs, 2554 differential expressed mRNAs and 116 differential expressed miRNAs were finally identified. Afterwards, a lncRNA mediated ceRNA network was constructed, according to the interactions among 544 pairs of DElncRNA-DEmiRNA relationships and 47 pairs of DEmiRNA-DEmRNA relationships. As for the survival analyses, we found 10 DElncRNAs, 25 DEmRNAs and 7 miRNAs have statistically prognostic significance for overall survival, respectively. CONCLUSIONS: This study provides meaningful information for deeper understanding the underlying molecular mechanism of lung adenocarcinoma and for evaluating prognosis, which could monitor recurrence, guide clinical treatment drugs and subsequent related researches.

The feasibility of navigation bronchoscopy-guided pulmonary microcoil localization of small pulmonary nodules prior to thoracoscopic surgery.

BACKGROUND: Accurate preoperative localization of small pulmonary nodules facilitates the rapid and precise video-assisted thoracoscopic surgery (VATS). This study aims to evaluate the feasibility, safety, and efficacy of navigation bronchoscopy-guided pulmonary microcoil placement for preoperative pulmonary nodule localization. METHODS: Twelve lung lesions were simulated by mixing lipiodol in three porcine models. After 1 week, two microcoils per lesion were deployed under bronchoscopic guidance. Computed tomography scans were then performed 1 day, 1 week, 2 weeks, and 4 weeks after the deployment to assess the position of the microcoils relative to the lesions. Surgical resection of the simulated lesions was performed under fluoroscopy 5 weeks after the deployment and the accuracy, stability, and associated complications of the microcoil localization were evaluated. Following this, an exploratory clinical study was conducted on three patients with pure ground-glass pulmonary nodules. RESULTS: The mean diameter of the twelve simulated lung lesions was 9.55±2.36 mm, and the mean distance from the pleura to the lesions was 8.29±2.99 mm. Twenty-four pulmonary microcoils were implanted in the bronchi surrounding the lesions. Four weeks later, the mean distance between the microcoils and the center of the lesions was 16.12±8.97 mm and the average migration of the microcoils relative to the baseline position (1 day after implantation) was 3.48±4.56 mm. All microcoils and target lesions were successfully resected in both the animal experiment and clinical study and no complications, such as pneumothorax, were observed during marker implantation or postoperative follow-up. CONCLUSIONS: The preoperative localization of pulmonary nodules by navigation bronchoscopy-guided microcoil placement is a safe, stable, and effective technique with minimal complication risk. This procedure can assist subsequent thoracoscopic resection.

Surgery performed at night by continuously working surgeons contributes to a higher incidence of intraoperative complications in video-assisted thoracoscopic pulmonary resection: a large monocentric retrospective study.

OBJECTIVES: Our goal was to assess the influence of working hours and working at night on intraoperative complications on surgeons conducting video-assisted pulmonary resections. METHODS: We identified all patients who underwent video-assisted thoracoscopic surgery (VATS) in Shanghai Chest Hospital from January 2015 to April 2017. Univariable and multivariable logistic analyses were used to analyse independent risk factors for intraoperative complications. A 1:4 propensity score matching analysis was conducted to verify those results. RESULTS: A total of 15 767 patients who underwent VATS pulmonary resection were included in this study. Among them, 15 280 patients (96.1%) were operated on during daytime working hours and 487 (3.1%) at night. A total of 203 (1.3%) intraoperative complications occurred. Vascular injury was the main cause of intraoperative complications, accounting for 92.1% (187/203). Multivariable logistic regression indicated that age [odds ratio (OR) = 1.68, 95% confidence interval (CI) 1.43-1.98; P < 0.001], gender (OR = 1.71, 95% CI 1.26-2.32; P = 0.001), surgical experience (OR = 2.07, 95% CI 1.56-2.75; P < 0.001), type of surgery (OR = 0.31, 95% CI 0.20-0.49; P < 0.001) and operative periods (OR = 2.69, 95% CI 1.61-4.86; P < 0.001) were independent predictors for intraoperative complications. The incidence of intraoperative complications during night-time surgery was significantly higher than that during daytime working hours. A 1:4 propensity score matching-based results verification showed that night-time surgery was still an independent risk factor after propensity score matching (OR = 2.76, 95% CI 1.47-5.15; P = 0.002). CONCLUSIONS: The incidence of intraoperative complications from VATS pulmonary resection performed during night hours was significantly higher than that performed during working hours. In the present labour environment, thoracic surgeons should avoid night-time surgery whenever possible.

HDAC6-specific inhibitor suppresses Th17 cell function via the HIF-1α pathway in acute lung allograft rejection in mice.

Background: Previous animal experiments and clinical studies indicated the critical role of Th17 cells in lung transplant rejection. Therefore, the downregulation of Th17 cell function in lung transplant recipients is of great interest. Methods: We established an orthotopic mouse lung transplantation model to investigate the role of histone deacetylase 6-specific inhibitor (HDAC6i), Tubastatin A, in the suppression of Th17 cells and attenuation of pathologic lesions in lung allografts. Moreover, mechanism studies were conducted in vitro. Results: Tubastatin A downregulated Th17 cell function in acute lung allograft rejection, prolonged the survival of lung allografts, and attenuated acute rejection by suppressing Th17 cell accumulation. Consistently, exogenous IL-17A supplementation eliminated the protective effect of Tubastatin A. Also, hypoxia-inducible factor-1α (HIF-1α) was overexpressed in a lung transplantation mouse model. HIF-1α deficiency suppressed Th17 cell function and attenuated lung allograft rejection by downregulating retinoic acid-related orphan receptor γt (ROR γt) expression. We showed that HDAC6i downregulated HIF-1α transcriptional activity under Th17-skewing conditions in vitro and promoted HIF-1α protein degradation in lung allografts. HDAC6i did not affect the suppression of HIF-1α-/- naïve CD4+ T cell differentiation into Th17 cell and attenuation of acute lung allograft rejection in HIF-1α-deficient recipient mice. Conclusion: These findings suggest that Tubastatin A downregulates Th17 cell function and suppresses acute lung allograft rejection, at least partially, via the HIF-1α/ RORγt pathway.