High glucose enhances the aggressiveness of lung adenocarcinoma via activating epidermal growth factor receptor/signal transducer and activator of transcription 3 pathways.

Epidemiological studies revealed hyperglycemia as a poor prognostic factor for lung adenocarcinoma with unclear molecular mechanisms. The present study thus aimed to investigate the effects of high glucose on the progression of lung adenocarcinoma and its underlying mechanisms. Lung adenocarcinoma cell lines, A549 and RERF-LC-KJ, were cultured in 5.6 mM glucose (normal glucose; NG) or 25 mM glucose (high glucose; HG) resembling euglycemia and hyperglycemia. Cells were examined for proliferation by the MTT assay, and migration-invasion using Transwell. The expressions of signaling proteins in epidermal growth factor receptor (EGFR) pathways and their downstream targets were investigated using Western blots. The effects of diabetes mellitus (DM) and hyperglycemia on lung adenocarcinoma growth in vivo were studied in streptozotocin-induced diabetic BALB/cAJcl-Nu/Nu mice and their nondiabetic counterparts. High glucose significantly promoted proliferation, migration, and invasion of lung adenocarcinoma cells compared with those in normal glucose (P<.05). Western blot analyses showed the increased ratio of pEGFR/EGFR in cells cultured in high glucose and subsequently activated the signal transducer and activator of transcription 3 (STAT3). Epithelial-mesenchymal (EMT) markers were also altered in lung adenocarcinoma cells in high glucose conditions, corresponding with increased migration and invasion abilities. Erlotinib, an EGFR inhibitor, significantly reversed high glucose-induced aggressive phenotypes confirming high glucose-enhancing lung adenocarcinoma progression via the activation of EGFR. DM and hyperglycemia also promoted the growth of lung adenocarcinoma xenografts in vivo in which erlotinib significantly suppressed the growth of tumors (P<.05) suggesting EGFR inhibitor as an effective therapeutic agent for lung adenocarcinoma with DM.

Powered stapling system with gripping surface technology for pulmonary resection of lung cancer: real-world clinical effectiveness.

OBJECTIVES: Surgical lung resection involves a critical task of stapled ligation and transection of major vascular structures and tissue, which may lead to bleeding and complications. A newer powered stapling system with Gripping Surface Technology (GST) was introduced to account for tissue movements. This study aimed to examine the real-world effectiveness of GST system on intraoperative and postoperative outcomes of pulmonary resection. METHODS: A retrospective analysis was conducted using the electronic medical records of Sichuan Provincial People's Hospital between July 2020 and March 2021 in China. Patients who underwent their first procedures of single-port lobectomy or multi-port segmentectomy by video-assisted thoracoscopic surgery were identified and grouped as GST group or manual stapler group (manual group) by the stapler types. The intraoperative outcomes such as bleeding rate, blood loss volume, and intervention rate at the staple line (including intraoperative pressure, suture, and electrocoagulation) were documented by trained nurses during the surgery. Propensity score matching was performed between the two groups, controlling forage, BMI, smoking history, history of surgery, complications, and level of complexity of pneumonectomy. RESULTS: A total of 108 matched patients were included in the analysis (54 in the GST group and 54 in the manual group). GST group had lower risks for intraoperative bleeding (22.8% vs 51.9%; p = 0.003) and intraoperative interventions (31.5% vs 55.6%; p = 0.02), compared to the manual group. A decrease in the intraoperative blood loss was observed in the GST group, but not statistically significant (134.39 ± 52.82 ml vs 158.11 ± 73.14 ml, p = 0.102). The use of NEOVEIL (reinforcement material to prevent air leakage from the staple line) intraoperatively during surgery was significantly lower in the GST group (24.1%) than in the manual group (50%, p = 0.01). CONCLUSION: The GST system was associated with better intraoperative outcomes in clinical practice in China.

Deep learning reveals cuproptosis features assist in predict prognosis and guide immunotherapy in lung adenocarcinoma.

Background: Cuproptosis is a recently found non-apoptotic cell death type that holds promise as an emerging therapeutic modality in lung adenocarcinoma (LUAD) patients who develop resistance to radiotherapy and chemotherapy. However, the Cuproptosis' role in the onset and progression of LUAD remains unclear. Methods: Cuproptosis-related genes (CRGs) were identified by a co-expression network approach based on LUAD cell line data from radiotherapy, and a robust risk model was developed using deep learning techniques based on prognostic CRGs and explored the value of deep learning models systematically for clinical applications, functional enrichment analysis, immune infiltration analysis, and genomic variation analysis. Results: A three-layer artificial neural network risk model was constructed based on 15 independent prognostic radiotherapy-related CRGs. The risk model was observed as a robust independent prognostic factor for LUAD in the training as well as three external validation cohorts. The patients present in the low-risk group were found to have immune "hot" tumors exhibiting anticancer activity, whereas the high-risk group patients had immune "cold" tumors with active metabolism and proliferation. The high-risk group patients were more sensitive to chemotherapy whereas the low-risk group patients were more sensitive to immunotherapy. Genomic variants did not vary considerably among both groups of patients. Conclusion: Our findings advance the understanding of cuproptosis and offer fresh perspectives on the clinical management and precision therapy of LUAD.

Cyclin-Dependent Kinase 4/6 Inhibitors: A Potential Breakthrough Therapy for Malignancies of Gastrointestinal Tract.

Cancer is the leading cause of death worldwide for which effective treatments remain limited. This article aimed to critically review and discuss the potential of targeting cell cycle machineries as a vital tool for cancer treatment. Cyclin dependent kinase (CDK) 4/6 inhibitors were originally approved by the United State Food and Drug Administration (US FDA) for advanced-stage breast cancer treatment. The nearly double-prolonged survival time in patients who received CDK4/6 inhibitors are superior to the conventional chemotherapy or endocrine therapy alone and, thus, these medications have been designated a breakthrough therapy by the US FDA. The requirement of CDK4/6 in the progression of cancer cells, but probably dispensable in normal cells, makes CDK4/6 a popular target for cancer treatment. The effects of CDK4/6 inhibitors in cancer may also involve the tumor microenvironment in which the therapeutic effects are synergistically pronounced. These emerging roles, hence, prompt investigations regarding their therapeutic potential in other cancers, including gastrointestinal cancer. Many preclinical and clinical studies of CDK4/6 inhibitors in gastrointestinal cancers are underway and, as a result, several new potentials are gradually reported. Contrariwise, the primary effect of this drug group is arresting the cell cycle rather than inducing cell death. The efficacy of using CDK4/6 inhibitors as a single regimen in clinical practice is then limited. In this article, the effects of CDK4/6 inhibitors on the progression of gastrointestinal cancers, at both preclinical and clinical levels are reviewed. The future directions for research and the possibility of CDK4/6 inhibitors being "breakthrough therapy" for gastrointestinal cancers are also discussed.

Inferior pulmonary ligament approach and/or interlobar fissure approach for posterior and/or lateral basal segment resection: a case-series of 31 patients.

Background: Thoracoscopic posterior and/ or lateral basal segment resection is a major difficult segmentectomy for thoracic surgeons, because of its high surgical difficulty and high incidence of postoperative complications. Here we describe the surgical procedure and techniques of the transpulmonary ligament approach and/or interlobar fissure approach for the thoracoscopic posterior and/or lateral basal segment resection. Methods: The clinical data of 31 patients who underwent thoracoscopic posterior and/or lateral basal segment resection from January 2020 to June 2022 were included. All patients underwent posterior and/or lateral basal segment resection via the inferior pulmonary ligament and/or interlobar fissure approach. Follow-up was continued to September 2022. Results: All patients including 7 males and 24 females, with a median age of 51 [31-62] years, completed the operation successfully. One patient was converted to lobectomy due to insufficient margins intraoperatively. Two patients were treated using the interlobar fissure approach, 25 patients were treated via the inferior pulmonary ligament approach, and four patients were treated by employing both methods. Also, 20 patients had a single lesion and 1 patient had two lesions. Eleven patients underwent surgical resections on other lesions when posterior and/or lateral basal segment lesions resected. The median operation time was 120 [50-290] minutes, the median intraoperative blood loss was 50 [10-100] mL, the median postoperative drainage time was 4 [2-10] days, and the median postoperative hospital stay was 4 [2-13] days. There was no perioperative death. Postoperative complications included five cases of persistent air leakage longer than 5 days (7, 7, 8, 9, and 10 days), and one patient developed a pulmonary infection and abnormal liver function after the operation. The median maximum diameter of the lesion was 0.8 [0.2-1.5] cm, lymph nodes were resected in a median of 8 [4-15] case. Conclusions: The approach of the inferior pulmonary ligament to resect posterior and/or the lateral basal segment can optimize the surgical procedure. The surgical trauma and postoperative complications are reduced, which is worthy of popularization and application.

An integrated model of FTO and METTL3 expression that predicts prognosis in lung squamous cell carcinoma patients.

BACKGROUND: Lung squamous cell carcinoma (LUSC) approximately accounts for a third of lung cancers. However, the role of N6-methyladenosine (m6A) in LUSC remains largely unknown according to previous studies. METHODS: In this study, we investigated the mutations, copy number variants (CNVs), expression of 20 m6A RNA methylation regulators, and clinical data from The Cancer Genome Atlas-LUSC (TCGA-LUSC). These data were used for the training cohort of screening potential biomarkers. The prognostic model of m6A RNA methylation regulators was constructed. A receiver operating characteristic (ROC) analysis was undertaken to determine the area under the curves (AUCs) (for 3- and 5-year survival) for the model. Additionally, the accuracy of the two-gene model was confirmed with external data verifications. Combined two-gene model and clinincal information were performed to construct a nomogram to predict patient's prognostic risk assessment. RESULTS: Fat mass- and obesity-associated protein (FTO) and methyltransferase-like 3 (METTL3) were identified as potential prognostic biomarkers to evaluate benign and malignant tumors and prognosticate. The following prognostic model of m6A RNA methylation regulators was constructed: risk score = 0.162 × FTO - 0.069 × METTL3. Patients in low-risk group [median overall survival (mOS), 43.4 months] had longer survival than those with high-risk (mOS, 67.3 months) with P=0.0023. The smoking grade and risk score could be independent prognostic factors (P=0.00098 and P=0.0014, respectively). Ultimately, a nomogram was developed to assist clinicians to predict clinical outcomes. CONCLUSIONS: FTO and METTL3 are potential prognostic biomarkers of LUSC. The two-gene model's use of prognostic risk scores may provide guidance in the selection of therapeutic strategies.

Analysis of risk factors of acute kidney injury in perioperative patients after lung transplantation.

BACKGROUND: This study sought to examine the factors influencing acute kidney injury (AKI) in the perioperative period after lung transplantation (LT). METHODS: We collected and analyzed the clinical data of 25 patients, who were diagnosed with AKI in the perioperative period after LT at Sichuan Provincial People's Hospital from July 1, 2018 to June 30, 2020. Based on the clinical outcomes, the patients were divided into an AKI group and a non-AKI group. Differences between the two groups were compared, including differences in mechanical ventilation (MV) time and intensive care unit (ICU) stay time, the mode of transplantation, the total amount of dehydration in the first week after surgery, use of potential kidney damaging drugs, and whether extracorporeal membrane oxygenation (ECMO) was used. RESULTS: Nineteen patients (76%) were diagnosed with AKI in the perioperative period after LT. There were no statistically significant differences between the two groups in terms of basic information, the mode of transplantation, the total amount of dehydration in the first week after surgery, the daily dose of tacrolimus, whether ganciclovir was used, whether voriconazole was used, whether ECMO was used, and mortality (P>0.05). However, the MV time and ICU stay time of the AKI group was longer than that of the non-AKI group (P=0.006, 0.011, respectively). Analysis within the group shows there were no significant differences in terms of mortality, the MV time, and the ICU stay time between the AKI stage two group and the AKI stage three group (all P>0.05). A multi-factor analysis was conducted in which AKI was the dependent variable, whether an amount of dehydration greater than 2,000 mL, a body mass index (BMI) greater than 18, and the use of ganciclovir and voriconazole had been examined as an independent variable; however, none of these were found to be risk factors associated with AKI. CONCLUSIONS: The incidence of AKI in the perioperative period after LT is high. AKI in the perioperative period after LT prolonged patients' MV time and ICU stay time. In the perioperative management of LT, it is necessary to consider kidney protection to reduce the risk of AKI.

Physiological and quantitative proteomic analysis of NtPRX63-overexpressing tobacco plants revealed that NtPRX63 functions in plant salt resistance.

High salinity is harmful to crop yield and productivity. Peroxidases (PRXs) play crucial roles in H2O2 scavenging. In our previous study, PRX63 significantly upregulated in tobacco plants under salt stress. Thus, in order to understand the function of PRX63 in tobacco salt response, we overexpressed this gene in tobacco (Nicotiana tabacum L.), investigated the morphological, physiological and proteomic profiles of NtPRX63-overexpressing tobacco transgenic lines and wild type. The results showed that, compared with the wild type, the transgenic tobacco plants presented enhanced salt tolerance and displayed lower ROS (reactive oxygen species), malondialdehyde (MDA) and Na+ contents; higher biomass, potassium content, soluble sugar content, and peroxidase activity; and higher expression levels of NtSOD, NtPOD and NtCAT. Protein abundance analysis revealed 123 differentially expressed proteins between the transgenic and wild-type plants. These proteins were functionally classified into 18 categories and are involved in 41 metabolic pathways. Furthermore, among the 123 proteins, eight proteins involved in the ROS-scavenging system, 12 involved in photosynthesis and energy metabolism processes, two stress response proteins, one signal transduction protein and one disulfide isomerase were significantly upregulated. Furthermore, three novel proteins that may be involved in the plant salt response were also identified. The results of our study indicate that an enhanced ROS-scavenging ability, together with the expression of proteins related to energy mobilization and the stress response, functions in the confirmed salt resistance of transgenic tobacco plants. Our data provide valuable information for research on the function of NtPRX63 in tobacco in response to abiotic stress.

Quantitative Proteomic Analysis of Alligator Weed Leaves Reveals That Cationic Peroxidase 1 Plays Vital Roles in the Potassium Deficiency Stress Response.

Alligator weed is reported to have a strong ability to adapt to potassium deficiency (LK) stress. Leaves are the primary organs responsible for photosynthesis of plants. However, quantitative proteomic changes in alligator weed leaves in response to LK stress are largely unknown. In this study, we investigated the physiological and proteomic changes in leaves of alligator weed under LK stress. We found that chloroplast and mesophyll cell contents in palisade tissue increased, and that the total chlorophyll content, superoxide dismutase (SOD) activity and net photosynthetic rate (PN) increased after 15 day of LK treatment, but the soluble protein content decreased. Quantitative proteomic analysis suggested that a total of 119 proteins were differentially abundant proteins (DAPs). KEGG analysis suggested that most represented DAPs were associated with secondary metabolism, the stress response, photosynthesis, protein synthesis, and degradation pathway. The proteomic results were verified using parallel reaction monitoring mass spectrometry (PRM-MS) analysis and quantitative real-time PCR (qRT-PCR)assays. Additional research suggested that overexpression of cationic peroxidase 1 of alligator weed (ApCPX1) in tobacco increased LK tolerance. The seed germination rate, peroxidase (POD) activity, and K+ content increased, and the hydrogen peroxide (H2O2) content decreased in the three transgenic tobacco lines after LK stress. The number of root hairs of the transgenic line was significantly higher than that of WT, and net K efflux rates were severely decreased in the transgenic line under LK stress. These results confirmed that ApCPX1 played positive roles in low-K+ signal sensing. These results provide valuable information on the adaptive mechanisms in leaves of alligator weed under LK stress and will help identify vital functional genes to apply to the molecular breeding of LK-tolerant plants in the future.

Physiology and proteomic analysis reveals root, stem and leaf responses to potassium deficiency stress in alligator weed.

Alligator weed is reported to have a strong ability to adapt to potassium deficiency stress. Proteomic changes in response to this stress are largely unknown in alligator weed seedlings. In this study, we performed physiological and comparative proteomics of alligator weed seedlings between normal growth (CK) and potassium deficiency (LK) stress using 2-DE techniques, including root, stem and leaf tissues. Seedling height, soluble sugar content, PGK activity and H2O2 contents were significantly altered after 15 d of LK treatment. A total of 206 differentially expressed proteins (DEPs) were identified. There were 72 DEPs in the root, 79 in the stem, and 55 in the leaves. The proteomic results were verified using western blot and qRT-PCR assays. The most represented KEGG pathway was "Carbohydrate and energy metabolism" in the three samples. The "Protein degradation" pathway only existed in the stem and root, and the "Cell cycle" pathway only existed in the root. Protein-protein interaction analysis demonstrated that the interacting proteins detected were the most common in the stem, with 18 proteins. Our study highlights protein changes in alligator weed seedling under LK stress and provides new information on the comprehensive analysis of the protein network in plant potassium nutrition.

[MRTF-A Regulates the Proliferation and Migration of Non-small Cell Lung Cancer Cells of A549 through HOTAIR].

BACKGROUND: Non-small cell lung cancer (NSCLC) is a kind of lung cancer, because its high incidence has been concerned. Therefore, it has great significance to reveal the pathogenesis of NSCLC. As a transcriptional regulatory factor, MATF-A plays an important role in the development of multiple tumors, can regulate the migration process of a variety of tumor cells. HOTAIR is a long non-coding RNA (LncRNA) found in recent years, which expresses abnormally in multiple tumors and is involved in the proliferation and migration of multiple tumors. The aim of this study is to explore the role of MRTF-A through HOTAIR to regulate the proliferation and migration of NSCLC cell A549 cell. METHODS: We constructed the overexpression plasmid and interfering plasmid of MRTF-A, and detected the effect of MRTF-A on the proliferation and migration of A549 cells by CCK8 and wound healing methods respectively. Then, we designed the siRNA of HOTAIR to detect its effect on the proliferation and migration of A549 cells. Through qRT-PCR, we detected the effect of MRTF-A on HOTAIR expression. Finally, we constructed HOTAIR's promoter, and detect the effect of MRTF-A on HOTAIR promoter activity by luciferase reporter gene test. RESULTS: Overexpression of MRTF-A promotes the proliferation and migration of A549 cells, while silent MRTF-A inhibits its proliferation and migration. Next, we found that interfered HOTAIR expression inhibited the proliferation of A549 cells. We found that MRTF-A could influence the expression of HOTAIR and regulate the activity of HOTAIR promoter. CONCLUSIONS: MRTF-A regulates the proliferation and migration of A549 cell through HOTAIR.

Physiological Analysis and Proteome Quantification of Alligator Weed Stems in Response to Potassium Deficiency Stress.

The macronutrient potassium is essential to plant growth, development and stress response. Alligator weed (Alternanthera philoxeroides) has a high tolerance to potassium deficiency (LK) stress. The stem is the primary organ responsible for transporting molecules from the underground root system to the aboveground parts of the plant. However, proteomic changes in response to LK stress are largely unknown in alligator weed stems. In this study, we investigated the physiological and proteomic changes in alligator weed stems under LK stress. First, the chlorophyll and soluble protein content and SOD and POD activity were significantly altered after 15 days of LK treatment. The quantitative proteomic analysis suggested that a total of 296 proteins were differentially abundant proteins (DAPs). The functional annotation analysis revealed that LK stress elicited complex proteomic alterations that were involved in oxidative phosphorylation, plant-pathogen interactions, glycolysis/gluconeogenesis, sugar metabolism, and transport in stems. The subcellular locations analysis suggested 104 proteins showed chloroplastic localization, 81 proteins showed cytoplasmic localization and 40 showed nuclear localization. The protein⁻protein interaction analysis revealed that 56 proteins were involved in the interaction network, including 9 proteins involved in the ribosome network and 9 in the oxidative phosphorylation network. Additionally, the expressed changes of 5 DAPs were similar between the proteomic quantification analysis and the PRM-MS analysis, and the expression levels of eight genes that encode DAPs were further verified using an RT-qPCR analysis. These results provide valuable information on the adaptive mechanisms in alligator weed stems under LK stress and facilitate the development of efficient strategies for genetically engineering potassium-tolerant crops.

Water stress enhances expression of genes encoding plastid terminal oxidase and key components of chlororespiration and alternative respiration in soybean seedlings.

Plastid terminal oxidase (PTOX) is a plastid-localized plastoquinone (PQ) oxidase in plants. It functions as the terminal oxidase of chlororespiration, and has the potential ability to regulate the redox state of the PQ pool. Expression of the PTOX gene was up-regulated in soybean seedlings after exposure to water deficit stress for 6 h. Concomitantly expression of the NDH-H gene, encoding a component of the NADPH dehydrogenase (NDH) complex which is a key component of both chlororespiration and NDH-dependent cyclic electron transfer (CET), was also up-regulated. Transcript levels of the proton gradient regulation (PGR5) gene, which encodes an essential component of the PGR5-dependent CET, were not affected by water stress, while the expression of the alternative oxidase (AOX1) gene, which encodes a terminal oxidase of alternative respiration in mitochondria, was also up-regulated under water stress. Therefore, our results indicate that water stress induced the up-regulation of genes encoding key components of chlororespiration and alternative respiration. Transcript levels of the AOX1 gene began to increase in response to water stress before those of PTOX suggesting that alternative respiration may react faster to water stress than chlororespiration.

Combined thoracoscopic and laparoscopic minimally invasive esophagectomy.

With the improvement in thoracoscopic and laparoscopic surgery, thoracoscopic and laparoscopic esophagectomy (TLE), a minimally invasive approach, has attracted increasing attention as an alternative to open three-field esophagectomy. From June 2012 to October 2013, 90 patients underwent laparoscopic and thoracoscopic resection of esophageal carcinoma in our department. The VATS esophagectomy technique described here is the approach currently employed in the department of thoracic surgery at Sichuan Provincial People's Hospital of China.