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.

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.

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.

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.

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.

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.

G-MDSCs-Derived Exosomal miRNA-143-3p Promotes Proliferation via Targeting of ITM2B in Lung Cancer.

BACKGROUND: The immune environment of lung cancer is complex, and the critical immune factors that promote lung cancer progression need to be explored. Granulocytic myeloid-derived suppressor cells (G-MDSCs) are regarded as immune suppressing cells. However, they also promote tumor progression through other ways, which needs to be explored further. Therefore, we sought to study the regulatory mechanisms underlying the cancer promoting function of G-MDSCs in lung cancer. METHODS: G-MDSCs were isolated from lung cancer tissues using flow cytometry. Exosomes were separated from the G-MDSCs supernatant by ultracentrifugation and verified using flow cytometry, Western blot, and transmission electron microscopy (TEM). RNA sequencing was used to identify the differential miRNAs and genes. Real-time quantitative real-time PCR (RT-qPCR) confirmed these results. The proliferation rate was assessed using the CCK-8 assay. Lentiviral vectors were used to alter the expression of the miRNAs and genes to analyze their effects on lung cancer progression. RESULTS: G-MDSCs secreted more exosomes in the lung cancer tissues, which promoted cancer progression by accelerating proliferation. Micro RNA-143-3p (miR-143-3p) increased in G-MDSCs derived exosomes and downregulated integral membrane protein 2B (ITM2B) by targeting the 3'-untranslated region (UTR) region. Overexpression of miR-143-3p enhanced proliferation by inhibiting transcription of ITM2B to activate the PI3K/Akt signaling pathway, which can be blocked by deguelin. This phenomenon was further confirmed by accelerated tumor growth and worse prognosis in mice. CONCLUSION: The key findings of this study highlight the potential of the G-MDSC-derived exosomes and the miR-143-3p/ITM2B axis as therapeutic targets and clinical indicators of 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.

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.

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.

Survival analyses and immunohistochemical study of primary signet ring cell carcinoma of the lung adenocarcinoma.

BACKGROUND: The clinicopathological features, immunohistochemical (IHC) characteristics and survival outcomes of primary lung adenocarcinoma with signet ring cells (LAdSRCs) component were analyzed. METHODS: A retrospective analysis of primary LAdSRCs consecutively collected from 2010 to 2014 was performed and compared with lung adenocarcinoma (ADC). We investigated the survival outcomes and the expression of cytokeratin 7 (CK7), CK20, thyroid transcription factor-1 (TTF-1) and villin in these 35 cases of primary LAdSRCs. RESULTS: We identified 1,715 patients in total, 35 (2.0%) of whom had signet ring cell (SRC) component. Excepting for 2 cases without IHC information, a CK profile (CK7+/CK20-) were identified in 69.7% (23/33) of LAdSRC; 22 (66.7%) of 33 LAdSRCs exhibited positive TTF-1 expression, but no other TTF-1 positive was found in signet ring cell carcinoma (SRCC) of other organs, and villin was not identified in these 33 cases. Age, sex, tumor size, surgery resection, smoking history, tumor stage, nodal stage, predominant histology subtype, pathology and adjuvant therapy were all significant predictors of relapse-free survival (RFS) and overall survival (OS) in univariable analysis. Compared to lung ADC, the lung ADCs with SRC component had an inferior prognosis. In multivariable analysis, sex, tumor size, tumor stage, nodal stage, predominant histology subtype and adjuvant therapy were still significant predictors for both RFS and OS while the pathology [RFS: hazard ratio (HR), 0.918; 95% confidence interval (CI), 0.581-1.492; P=0.712; OS: HR, 1.392; 95% CI, 0.820-2.364; P=0.221, respectively] was not. CONCLUSIONS: Univariable analysis revealed primary LAdSRC is a rare ADC subtype with a worse prognosis compared with ADC. The high percentage expression of TTF-1 and immunostaining profiles CK7+/CK20- in primary LAdSRC is significant to identify the source of SRCCs.

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.

Identification of Common Genes and Pathways in Eight Fibrosis Diseases.

Acute and chronic inflammation often leads to fibrosis, which is also the common and final pathological outcome of chronic inflammatory diseases. To explore the common genes and pathogenic pathways among different fibrotic diseases, we collected all the reported genes of the eight fibrotic diseases: eye fibrosis, heart fibrosis, hepatic fibrosis, intestinal fibrosis, lung fibrosis, pancreas fibrosis, renal fibrosis, and skin fibrosis. We calculated the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment scores of all fibrotic disease genes. Each gene was encoded using KEGG and GO enrichment scores, which reflected how much a gene can affect this function. For each fibrotic disease, by comparing the KEGG and GO enrichment scores between reported disease genes and other genes using the Monte Carlo feature selection (MCFS) method, the key KEGG and GO features were identified. We compared the gene overlaps among eight fibrotic diseases and connective tissue growth factor (CTGF) was finally identified as the common key molecule. The key KEGG and GO features of the eight fibrotic diseases were all screened by MCFS method. Moreover, we interestingly found overlaps of pathways between renal fibrosis and skin fibrosis, such as GO:1901890-positive regulation of cell junction assembly, as well as common regulatory genes, such as CTGF, which is the key molecule regulating fibrogenesis. We hope to offer a new insight into the cellular and molecular mechanisms underlying fibrosis and therefore help leading to the development of new drugs, which specifically delay or even improve the symptoms of fibrosis.

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.

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.

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.

Knockdown of TRIM65 inhibits autophagy and cisplatin resistance in A549/DDP cells by regulating miR-138-5p/ATG7.

Cisplatin resistance is the main cause of treatment failure in patients with non-small-cell lung cancer (NSCLC). Autophagy is a key mechanism of resistance to chemotherapy. Given that tripartite motif (TRIM)-containing proteins are involved in the regulation of autophagy and chemoresistance, we aimed to study the functions of TRIM protein members in autophagy-mediated chemoresistance of NSCLC. We found that TRIM65 was significantly increased in cisplatin-resistant NSCLC cell line (A549/DDP) as compared to the parental cell line (A549). Knockdown of TRIM65 can enhance cisplatin-induced apoptosis and inhibit autophagy in A549/DDP cells, as indicated by Annexin V/PI staining, caspase3 activity test, and LC3-II immunofluorescence staining. Additionally, knockdown of TRIM65 significantly decreased the expression of an important autophagy mediator, ATG7, which was a potential target of miR-138-5p. miR-138-5p inhibitor significantly abolished the effects of TRIM65 knockdown on autophagy and cisplatin-induced apoptosis. Moreover, TRIM65 induced the ubiquitination and degradation of TNRC6A, resulting in the suppressed expression of miR-138-5p. TRIM65 knockdown inhibited the growth of tumors derived from A549/DDP cells. Furthermore, cisplatin-resistant NSCLC tissues displayed higher expression of TRIM65 mRNA and lower expression of miR-138-5p as compared to cisplatin non-resistant ones. miR-138-5p expression was negatively correlated with TRIM65 mRNA in NSCLC tissues. Collectively, the present study indicates that TRIM65 knockdown attenuates autophagy and cisplatin resistance in A549/DDP cells via regulating miR-138-5p.

Prognosis analysis of lobectomy and sublobar resection in patients ≥75 years old with pathological stage I invasive lung adenocarcinoma of ≤3 cm: a propensity score matching-based analysis.

BACKGROUND: To determine the clinical prognosis after sublobectomy versus lobectomy in elderly patients ≥75 years old with stage I invasive lung adenocarcinoma ≤3 cm in size. METHODS: In patients ≥75 years old, 255 patients were diagnosed with stage I invasive lung adenocarcinoma ≤3 cm in size between 2010 and 2014 in Shanghai Chest Hospital, they were all treated with sublobectomy or lobectomy. Potential confounding factors that consisted in the baseline characteristics of these two groups was balanced by the method of propensity score matching (PSM). The stratified analysis was conducted to compare the relapse-free survival (RFS) and lung cancer special survival (LCSS) rates in the sublobectomy and lobectomy groups. RESULTS: As for the 255 patients, 112 cases conducted sublobectomy and 143 with lobectomy. Significant difference existed in RFS before (P=0.002) and after (P=0.010) PSM. Similarly, we still recognized significant difference in LCSS between the two groups before (log-rank P<0.001) or after (log-rank P=0.002) PSM. We still identified different RFS or LCSS rates between the stratified tumor size group and the stratified lymph node dissection group after adjustment of PSM. CONCLUSIONS: Lobectomy showed a survival advantage for sublobectomy for patient ≥75 years old with stage I lung adenocarcinoma ≤3 cm in size. Considering that lobectomy could get a better prognosis, it should be preferable for the treatment of patient ≥75 years old with stage I lung adenocarcinoma ≤3 cm in size.

Pulmonary retransplantation.

Lung retransplantation has developed quickly these years with the increasing number of primary lung transplantation. But the operative risk and survival of lung retransplantation was inferior to primary lung transplantation. The bronchiolitis obliterans syndrome (BOS), the early graft failure and the irreversible airway complications are the main causes for lung retransplantation. In this review, we give a general view of the history of lung retransplantation. And we reviewed the factor related to the prognosis of lung retransplantation according to the previous publications. There have been four lung retransplantation in Shanghai Chest Hospital till now. We shared our preliminary experience here.

Application of ex vivo lung perfusion (EVLP) in lung transplantation.

Lung transplantation was the ultimate method to treat the end-stage benign lung diseases. Although the lung transplantation has been increasing quickly in the past 30 years, the shortage of donor lung still limited its development. Ex vivo lung perfusion (EVLP) was a promising technique that could provide the platform to preserve, evaluate and repair the donor lung graft. In this article, we give a general review of the development of EVLP, its clinical application and the animal model. With the increasing experience of clinical EVLP, some pre-identified unsuitable donor lungs have been re-evaluated and accepted for transplantation. EVLP have now been considered to be an effective way to expand the donor pool.