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BACKGROUND: Uniportal video-assisted thoracoscopic surgery (VATS) segmentectomy is widely used in the field of thoracic surgery. However, anatomical variations in the bronchi and lung vessels may be critical obstacles during precise pulmonary segmentectomy. Thus, it is necessary to optimize uniportal VATS segmentectomy and to accurately identify the plane between lung segments by precisely transecting the bronchi and blood vessels of the lung segments. The indocyanine green fluorescence (ICGF)-based method has the potential to be a feasible and effective technique to facilitate the uniportal VATS segmentectomy. The present study aims at comparing the short-term outcomes of ICGF versus the traditional inflation-deflation method for uniportal VATS segmentectomy. METHODS: The perioperative clinical data in 200 consecutive patients undergoing uniportal VATS segmentectomy from December 2018 to August 2020 at Shanghai Chest Hospital were analyzed retrospectively. The targeted segment structures were identified and dissected precisely by using ICGF-based (N=100) or the traditional inflation-deflation (N=100) methods. The parameters of intraoperative blood loss and operation time, postoperative drainage volume, air leakage time, drainage tube retention time, length of hospital stay, and complications in the ICGF group were collected. Further, the operation time between the ICGF and the inflation-deflation groups was compared. The data summary and statistical analysis were performed by SPSS 19.0. P value

The thick filament is a key component of sarcomeres, the basic force-generating and load-bearing unit of striated muscle1. Mutations in thick filament proteins are associated with familial hypertrophic cardiomyopathy and other heart and muscle diseases2, 3. Despite this central importance for sarcomere force generation, it remains unclear how thick filaments are structurally organized and how its components interact with each other and with thin filaments to enable highly regulated muscle contraction. Here, we present the molecular architecture of native cardiac sarcomeres in the relaxed state, determined by electron cryo-tomography. Our reconstruction of the thick filament reveals the three-dimensional organization of myosin heads and tails, myosin-binding protein C (MyBP-C) and titin, elucidating the structural basis for their interaction during muscle contraction. The arrangement of myosin heads is variable depending on their position along the filament, suggesting that they have different capacities in terms of strain susceptibility and activation. Myosin tails exhibit a distinct arrangement and pattern of interactions. These are likely orchestrated by three alpha and three beta titin chains that are arranged like a spring, suggesting the existence of specialized roles of thick filament segments in length-dependent activation and contraction. Surprisingly, while the three titin alpha chains run along the entire length of the thick filament, titin beta does not. The structure also demonstrates that the C-terminal region of MyBP-C binds myosin tails and unexpectedly also directly interacts with the myosin heads, suggesting a previously undescribed direct role in the preservation of the myosin OFF state. Furthermore, we visualize how MyBP-C forms links between thin and thick filaments. These findings establish a robust groundwork for forthcoming research endeavors aiming to explore muscle disorders that involve sarcomeric structural components.

Muscles are essential for movement and heart function. Contraction and relaxation of muscles relies on the sliding of two types of filaments—the thin filament and the thick myosin filament. The thin filament is composed mainly of filamentous actin (F-actin), tropomyosin, and troponin. Additionally, several other proteins are involved in the contraction mechanism, and their malfunction can lead to diverse muscle diseases, such as cardiomyopathies. We review recent high-resolution structural data that explain the mechanism of action of muscle proteins at an unprecedented level of molecular detail. We focus on the molecular structures of the components of the thin and thick filaments and highlight the mechanisms underlying force generation through actin–myosin interactions, as well as Ca2+-dependent regulation via the dihydropyridine receptor, the ryanodine receptor, and troponin. We particularly emphasize the impact of cryo–electron microscopy and cryo–electron tomography in leading muscle research into a new era. Expected final online publication date for the Annual Review of Biochemistry, Volume 92 is June 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Disease-causing bacteria use a variety of secreted toxins to invade and subjugate their hosts. While the machinery responsible for secretion of many smaller toxins has already been established, it remains enigmatic for larger ones such as Tc toxins from human and insect pathogens, which approach the size of a prokaryotic ribosome. In the present study, we combine targeted genomic editing, proteomic profiling and cryo-electron tomography of the insect pathogenYersinia entomophagato reveal that a specialized subset of bacterial cells produces the Tc toxin YenTc as part of a complex toxin cocktail released into the environment by controlled cell lysis using a transcriptionally-coupled, pH-dependent type 10 secretion system (T10SS). Our results dissect the process of Tc toxin export by a T10SS in hitherto unprecedented detail, identifying that T10SSs operate via a previously unknown lytic mode of action, and establishing them as crucial players in the size-insensitive release of cytoplasmically folded toxins. With T10SSs directly embedded in Tc toxin operons of major human pathogens such asYersinia pestisandSalmonella enterica, we anticipate our findings to model an important aspect of pathogenesis in bacteria with a significant impact on global human health.

BACKGROUND: Carinal resection, performed in only a few high-volume centers, remains one of the most complicated and technically demanding surgeries. Few studies have examined the outcomes of carinal resection and reconstruction with complete pulmonary parenchyma preservation. METHODS: Patients who underwent isolated carinal resection and reconstruction at the Shanghai Chest Hospital between 2006 and 2020 were retrospectively reviewed. Clinicopathological, perioperative, and follow-up outcomes were analyzed. RESULTS: A total of 36 patients were included, including 19 men and 17 women. The average age was 50.7±14.8 years. Right posterolateral thoracotomy (n=33, 91.7%) and cross-field intubation during anastomosis (n=31, 86.1%) were selected for the majority of the carinal surgeries. The average intraoperative blood loss was 225.0 mL, and the mean operation duration was 196.1 minutes. Postoperative complications were observed in 14 patients (38.9%), including cicatricial stenosis (n=8, 22.2%), anastomotic fistula (n=3, 8.3%), air leak (n=1, 2.8%), cardiac arrhythmia (n=4, 11.1%), pneumonia (n=2, 5.6%), respiratory failure (n=1, 2.8%), and pulmonary embolism (n=1, 2.8%). There were 2 perioperative deaths (5.6%). Multivariate analysis revealed that being overweight was an independent favorable factor for postoperative complications [P=0.042, odds ratio (OR) =0.092, 95%, confidence interval (CI): 0.009–0.922]. Pathological diagnoses included squamous cell carcinoma (SCC) (n=12, 33.3%), adenoid cystic carcinoma (ACC) (n=15, 41.7%), mucoepidermoid carcinoma (MEC) (n=2, 5.6%), stricture (n=1, 2.8%), and other rare histological types. An R0 resection was achieved in 14 patients, while 21 patients (60.0%) had microscopically positive margins. Lymph node metastasis was confirmed in 6 patients (17.1%). Overall survival (OS) was 94.4% at 1 year and 79.4% at 5 years, with 107 months as the median survival time (95% CI: 64.0–150.0 months). All patients with negative margins remained alive during the follow-up period, while those who received R1 resections had much poorer survival rates due to tumor recurrence [P=0.042, hazard ratio (HR) =4.938, 95% CI: 1.062–22.950]. CONCLUSIONS: In selected patients, carinal resection and reconstruction with complete pulmonary parenchyma preservation was a feasible option to achieve an appreciable long-term survival at the risk of acceptable operative mortality and morbidity, particularly when complete resection with negative margins could be realized.

BACKGROUND: Airway management in tracheobronchial surgeries, especially carinal resection and reconstruction, remains one of the greatest challenges to thoracic surgeons. This study investigated the safety and effectiveness of venovenous extracorporeal membrane oxygenation (VV-ECMO) for respiratory support during tracheobronchial surgeries. METHODS: The data of patients who underwent VV-ECMO-assisted tracheobronchial surgeries at the Shanghai Chest Hospital from August 2006 to August 2021 were retrospectively reviewed. The clinicopathological, perioperative, and follow-up outcomes were analyzed. RESULTS: A total of 7 patients (4 males and 3 females) with a median age of 56 years (range, 11–70 years) were included in the study. The following tracheobronchial surgeries were conducted: carinal resection and reconstruction with complete pulmonary parenchyma preservation (n=4), left main bronchus and hemi-carinal sleeve resection (n=1), right upper sleeve lobectomy and hemi-carinal resection (n=1), and tracheal resection and reconstruction (n=1). The mean time on VV-ECMO was 167.7±65.8 min, and the mean operative time was 192.4±55.0 min. The average estimated blood loss was 271.4±125.4 mL. No perioperative death or reimplantation of VV-ECMO occurred. Postoperative complications were observed in 2 patients, including 1 case of respiratory failure due to preoperative severe chronic obstructive pulmonary disease (COPD) and 1 case of chylothorax. The median hospital stay was 11 days (range, 7–46 days). The median follow-up time was 30 months (range, 21–33 months). All the patients remained alive, and no postoperative readmission occurred during the follow-up period. CONCLUSIONS: VV-ECMO is a safe and feasible ventilation mode when intraoperative oxygen saturation cannot be well maintained during tracheobronchial surgery.

BACKGROUND: This study assessed the incidence and risk factors (RFs) of postoperative atrial fibrillation (POAF) and its impact on clinical outcomes in patients undergoing pneumonectomy for lung cancer. METHODS: Between 2013 and 2018, this monocentric retrospective study enrolled 324 consecutive pneumonectomy patients for primary lung cancer from our institution and 350 lobectomy and 349 segmentectomy cases matched by age, sex and body mass index (BMI). RF for POAF and postoperative death in pneumonectomy patients were assessed by logistic regression, and long-term outcomes after a median follow-up of 30 (range, 2–61) months by Cox proportional hazard model. Electrophysiology study (EPS) files of 30 AF patients with lung resection history were reviewed. RESULTS: POAF developed more often after pneumonectomy than lobectomy and segmentectomy (23.2% vs. 6.6% vs. 1.4%, respectively; P60 years and left atrial diameter (LAd) ≥35 mm as independent predictors. POAF, infection and hemorrhage were independent RFs for perioperative death after pneumonectomy; however, POAF was not RF for long-term death. Pulmonary vein (PV) trigger was identified in 60% (18/30) of AF patients with lung resection history, with stump PVs being more active than non-stump PVs (38.2% vs. 10.5%, P60 years and LAd ≥35 mm, was mostly solitary and possibly secondary to stump and non-stump PV triggers. POAF, along with infection and hemorrhage, was a RF for perioperative death.

OBJECTIVES The paediatric tracheobronchial inflammatory myofibroblastic tumour (IMT) is a rare disease. Whether limited surgical resection is a feasible surgical approach for these patients remains controversial. The objectives of this study were to report the long-term prognosis after limited surgical resections on paediatric tracheobronchial IMT and provide a surgical management strategy for this rare disease. METHODS Paediatric tracheobronchial IMT patients who underwent limited surgical resection from 2012 to 2020 were enrolled in this study. The clinical characteristics, course of treatment and long-term outcomes of all participants were collated. We presented the accumulated data and analysed the feasibility of limited surgical resection on the paediatric tracheobronchial IMT. RESULTS A total of 9 children with tracheobronchial IMTs were enrolled in our study. Cough and shortness of breath were the most common symptoms. All 9 participants underwent surgical treatment, including 2 tracheal reconstructions, 4 carinal reconstructions and 3 bronchial sleeve resections. Among the participants, 6/9 (66%) were positive for the anaplastic lymphoma receptor tyrosine kinase gene in terms of immunohistochemistry. None of the participants died of short-term complications. The follow-up period was 5.4 (range, 1.1–9.3) years, during which all participants remained well. CONCLUSIONS Limited surgical resection is preferred for paediatrics with tracheobronchial IMTs. Meanwhile, patients with complete resection have an excellent long-term prognosis.

Background Phrenic nerve injury (PNI) during lung cancer surgery, without apparent nerve section or damage, is still not well-studied. The aim of our study is to find an easy and objective way to evaluate a significant diaphragm elevation (SDE) suggestive of inadvertent PNI and its incidence and impact on lung cancer patients undergone video-assisted thoracoscopic surgery (VATS) lobectomy. Methods Extent of diaphragm elevation was first examined on chest X-ray in a cohort of patients with invasive thymoma in whom phrenic nerve was intentionally transected. The result was then used as the criterion to diagnose a SDE suggestive of PNI in another cohort of VATS lobectomy patients. Fluoroscopy test was used to validate the results. Spirometry test was repeated to evaluate pulmonary function loss after surgery. Results Diaphragm elevation was 24.24%±6.2% in 22 invasive thymoma-patients, with 30% elevation adopted as the criterion to diagnose SDE suggestive of PNI. In 753 VATS lobectomy patients, 56 (7.4%) were diagnosed of SDE. On Fluoroscopy test, diaphragm movement was significantly less in patients with diaphragm elevation >30% than those without (5.0 vs. 11.0 mm, P=0.003), together with a significantly smaller diaphragm movement ratio on the operation (OP) side than on the contralateral side (17% vs. 42%, P=0.018). Although no difference in postoperative complications was found, reduction in FEV1, FVC, and DLCO was significantly greater in patients with a SDE than those without (P=0.009). Conclusions Patients with more than 30% diaphragm elevation after VATS lobectomy is highly likely to have PNI and should undergo fluoroscopic validation. Inadvertent PNI during VATS lobectomy is an underestimated phenomenon and is associated with significantly greater loss of pulmonary function.

Rationale: Subsets of patients with early-stage lung adenocarcinoma (LUAD) have a poor post-surgical course after curative surgery. However, biomarkers stratifying this high-risk subset and molecular underpinnings underlying the aggressive phenotype remain unclear. Methods: We integrated bulk and single-cell transcriptomics, proteomics, secretome and spatial profiling of clinical early-stage LUAD samples to identify molecular underpinnings that promote the aggressive phenotype. Results: We identified and validated THBS2, at multi-omic levels, as a tumor size-independent biomarker that robustly predicted post-surgical survival in multiple independent clinical cohorts of early-stage LUAD. Furthermore, scRNA-seq data revealed that THBS2 is exclusively derived from a specific cancer-associated fibroblast (CAF) subset that is distinct from CAFs defined by classical markers. Interestingly, our data demonstrated that THBS2 was preferentially secreted via exosomes in early-stage LUAD tumors with high aggressiveness, and its levels in the peripheral plasma associated with short recurrence-free survival. Further characterization showed that THBS2-high early-stage LUAD was characterized by suppressed antitumor immunity. Specifically, beyond tumor cells, THBS2+ CAFs mainly interact with B and CD8+ T lymphocytes as well as macrophages within tumor microenvironment of early-stage LUAD, and THBS2-high LUAD was associated with decreased immune cell infiltrates but increased immune exhaustion marker. Clinically, high THBS2 expression predicted poor response to immunotherapies and short post-treatment survival of patients. Finally, THBS2 recombinant protein suppressed ex vivo T cells proliferation and promoted in vivo LUAD tumor growth and distant micro-metastasis. Conclusions: Our multi-level analyses uncovered tumor-specific THBS2+ CAFs as a key orchestrator promoting aggressiveness in early-stage LUAD.

In skeletal muscle, nebulin stabilizes and regulates the length of thin filaments, but the underlying mechanism remains nebulous. In this work, we used cryo–electron tomography and subtomogram averaging to reveal structures of native nebulin bound to thin filaments within intact sarcomeres. This in situ reconstruction provided high-resolution details of the interaction between nebulin and actin, demonstrating the stabilizing role of nebulin. Myosin bound to the thin filaments exhibited different conformations of the neck domain, highlighting its inherent structural variability in muscle. Unexpectedly, nebulin did not interact with myosin or tropomyosin, but it did interact with a troponin T linker through two potential binding motifs on nebulin, explaining its regulatory role. Our structures support the role of nebulin as a thin filament “molecular ruler” and provide a molecular basis for studying nemaline myopathies.

Background Locally-advanced lung squamous cell carcinoma represents a special subset that is challenging to resect completely with surgery alone. Immunotherapy has achieved great success in treating late-stage lung cancer. However, whether neoadjuvant immunotherapy can facilitate resection of initially locally-advanced and surgically-difficult locally-advanced lung squamous cell carcinoma remains to be investigated. Methods We retrospectively collected clinical records of locally-advanced lung squamous cell carcinoma patients who received neoadjuvant immunotherapy followed by surgery between 2018 and 2020 at a large academic thoracic cancer center. Results A total of 23 patients (22 males, 1 female) with locally-advanced locally-advanced lung squamous cell carcinoma were included, initially clinically staged at IIIA (16, 69.6%), IIIB (n=4, 17.4%), IIB (n=2, 8.7%) and IIIC (n=1, 4.3%). The median interval between final treatment to surgery was 36 days (range, 25-93 days), without treatment-related delay in surgery. The neoadjuvant treatment resulted in a high rate of radical resection (n=20, 87.0%). The final histopathological examination demonstrated 6 (26.1%) cases with pathological complete response and 8 (34.8%) with pathological major response. Comparing with the computed tomography scan-based response, we observed a very low consistency (weighted kappa =0.122, P=0.315) between the computed tomography scan-based and final pathological evaluation. The median follow-up time was 510 days (range, 217-920 days). At the end of the follow-up, 1 patient died. Conclusions Our findings showed the clinical promise of neoadjuvant immunotherapy plus surgery for locally-advanced lung squamous cell carcinoma. Computed tomography scan displays a poor role in assessing the resectability after neoadjuvant immunotherapy.

Nebulin is a major structural protein of skeletal sarcomeres and is essential for proper assembly and contraction of skeletal muscle1. It stabilises and regulates the length of thin filaments,2 but the structural mechanism remains nebulous. Using electron cryotomography and sub-tomogram averaging, we present the first structure of native nebulin bound to thin filaments within the A-band and I-band of intact sarcomeres. This in-situ reconstruction reveals unprecedented detail of interaction at pseudo-atomic resolution between nebulin and actin, providing the basis for understanding the structural and regulatory roles of nebulin. The position of nebulin on the thin filament indicates that there is no contact to tropomyosin or myosin, but an unexpected interaction with a troponin-T linker, possibly through two binding motifs on nebulin. In addition, our structure of myosin bound to the thin filaments reveals different conformations of the neck domain, both within the same sarcomere and when compared to purified structures, highlighting an inherent structural variability in muscle. We provide a complete description of cross-bridge formation on fully native, nebulin-containing thin filaments at near-atomic scale. Our structures establish the molecular basis for the role of nebulin as a thin filament “molecular ruler” and the impact of nemaline myopathies mutations that will aid future development of therapies.

The simulation results of the air flow around NACA 2412 airfoil in STARCCM+ are submitted. The 9 groups of comparative experiments were simulated at 9 angles of attack: 0, 10, 13, 14, 15, 16, 18, 20, 30 degrees. By analyzing the air flow near the upper surface airfoil, the relationship between the generation condition of separation vortex and the angle of attack and the law of change of angle of attack and lift force were obtained. Finding the range of angle of attack for the stall, the angle of attack with maximum lift effectiveness was determined.

BACKGROUND: Video-assisted thoracoscopic surgery (VATS) has been increasingly used in the treatment of lung cancers. But it is still unclear whether mediastinal lymph node dissection (LND) under VATS is safe and feasible. The aim of this study is to figure out whether LND by VATS is safe and feasible. METHODS: Consecutive patients with primary resectable lung cancers referred for lobectomy and LND or sampling by VATS between January 2012 and December 2016 were retrospectively reviewed. Clinicopathological characteristics and perioperative results were collected for statistical analysis. RESULTS: Seven-hundred and seventy-three VATS lobectomy patients were included in this study, 494 received LND and 279 received lymph node sampling (LNS). There were more male patients, higher pathological T and N stage in the LND group than in the LNS group. Multivariate analysis suggested that clinical N stage higher than cN0 category and LND were independent risk factors for finding pN2 diseases in all lung cancers, while higher than cN0 category, solid or micropapillary component, and LND were independently related to finding pN2 stage in adenocarcinomas. Propensity-score matching rendered 279 pairs of patients with no significant difference in age, gender, co-morbidity, tumor location, or T stage. Although the LND group had longer operation time (128 vs. 114 minutes, P

Cryo-electron tomography (cryo-ET) is an emerging technique to study the cellular architecture and the structure of proteins at high resolution in situ. Most biological specimens are too thick to be directly investigated and are therefore thinned by milling with a focused ion beam under cryogenic conditions (cryo-FIB). This procedure is prone to contaminations, which makes it a tedious process, often leading to suboptimal results. Here, we present new hardware that overcomes the current limitations. We developed a new glove box and a high vacuum cryo transfer system and installed a stage heater, a cryo-shield and a cryo-shutter in the FIB milling microscope. This reduces the ice contamination during the transfer and milling process and simplifies the handling of the sample. In addition, we tested a new software application that automates the key milling steps. Together, these improvements allow for high-quality, high-throughput cryo-FIB milling. This paves the way for new types of experiments, which have been previously considered infeasible.

BACKGROUND: Video-assisted thoracoscopic surgery (VATS) has been widely used in the lung resections. Reports regarding three-dimension (3D) single-port VATS are very limited. The purpose of this study is to evaluate the perioperative outcomes of 3D single-port VATS in a single medical center. METHODS: Totally 523 clinical stage I lung cancer patients underwent surgical resection through VATS operation between September 2016 and October 2017 in our single institution were retrospectively collected and 374 were enrolled. The comparison between 3D single-port VATS and conventional VATS (c-VATS), single-port VATS was conducted focusing on intraoperative and postoperative outcomes. Continuous and categorical variables were analyzed through SPSS software. RESULTS: The 3D singe-port VATS demonstrated no significant difference neither on the intraoperative outcomes including the operative time and the intraoperative blood loss nor the postoperative outcomes including the length of drainage duration and postoperative complications when against c-VATS and single-port VATS. Besides, 3D singe-port VATS elucidated comparable ability of lymph node dissection with c-VATS in subgroup analysis (P=0.192), both of which were better than single-port VATS group (P

Background There is a paucity of biomarkers that can predict the degree of pathological response [e.g., pathological complete response (pCR) or major response (pMR)] to immunotherapy. Neoadjuvant immunotherapy provides an ideal setting for exploring responsive biomarkers because the pathological responses can be directly and accurately evaluated. Methods We retrospectively collected the clinicopathological characteristics and treatment outcomes of non-small cell lung cancer (NSCLC) patients who received neoadjuvant immunotherapy or chemo-immunotherapy followed by surgery between 2018 and 2020 at a large academic thoracic cancer center. Clinicopathological factors associated with pathological response were analyzed. Results A total of 39 patients (35 males and 4 females) were included. The most common histological subtype was lung squamous cell carcinoma (LUSC) (n=28, 71.8%), followed by lung adenocarcinoma (LUAD) (n=11, 28.2%). After neoadjuvant treatment, computed tomography (CT) scan-based evaluation showed poor agreement with the postoperatively pathological examination (weighted kappa =0.0225; P=0.795), suggesting the poor performance of CT scans in evaluating the response to immunotherapy. Importantly, we found that the smoking signature displayed a better performance than programmed death-ligand 1 (PD-L1) expression in predicting the pathological response (area under the curve: 0.690 vs. 0.456; P=0.0259), which might have resulted from increased tumor mutational burden (TMB) and/or microsatellite instability (MSI) relating to smoking exposure. Conclusions These findings suggest that CT scan-based evaluation is not able to accurately reflect the pathological response to immunotherapy and that smoking signature is a superior marker to PD-L1 expression in predicting the benefit of immunotherapy in NSCLC patients.

Background Intrinsic or acquired resistance to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) is common, thus strategies for the management of EGFR-TKIs resistance are urgently required. Ferroptosis is a recently discovered form of cell death that has been implicated in tumorigenesis and resistance treatment. Accumulating evidence suggests that ferroptosis can be therapeutically exploited for the treatment of solid tumors; however, whether ferroptosis can be targeted to treat EGFR mutant lung cancer and/or overcome the resistance to EGFR-TKIs is still unknown. Methods The effect of ferroptosis inducers on a panel of EGFR mutant lung cancer cell lines, including those with EGFR-TKI intrinsic and acquired (generated by long-term exposure to the third-generation EGFR-TKI osimertinib), was determined using cytotoxicity assays. Further, drug candidates to enhance the effect of ferroptosis inducers were screened through implementing WGCNA (weighted gene co-expression network analysis) and CMAP (connectivity map) analysis. Flow cytometry-based apoptosis and lipid hydroperoxides measurement were used to evaluate the cell fates after treatment. Results Compared with EGFR-TKI-sensitive cells, those with intrinsic or acquired resistance to EGFR-TKI display high sensitivity to ferroptosis inducers. In addition, Vorinostat, a clinically used inhibitor targeting histone deacetylase, can robustly enhance the efficacy of ferroptosis inducers, leading to a dramatic increase of hydroperoxides in EGFR mutant lung cancer cells with intrinsic or acquired resistance to EGFR-TKI. Mechanistically, Vorinostat promotes ferroptosis via xCT downregulation. Conclusions Ferroptosis-inducing therapy shows promise in EGFR-activating mutant lung cancer cells that display intrinsic or acquired resistance to EGFR-TKI. Histone deacetylase inhibitor (HDACi) Vorinostat can further promote ferroptosis by inhibiting xCT expression.

Left-side secondary carina resection and reconstruction is a rare, complex procedure, performed just in a few specialized centers in a restricted group of patients. Few studies describe this technique and report its short and long-term results. We reviewed our experience to evaluate the perioperative and short-term outcomes of a very demanding surgery. We retrospectively collected the information of all the patients who underwent secondary carina resection and reconstruction for low-grade malignant bronchial tumors at our center. Between January 2012 and September 2018, 23 patients received surgery for low-grade malignant bronchial tumors. In all patients, a secondary carina resection and reconstruction with total lung parenchymal preservation was performed. The mean age was 44.5 ± 12.2 years. Pathologies included adenoid cystic carcinoma in ten patients, carcinoid in 7 (6 typical and 1 atypical), mucoepidermoid carcinoma in 4, myoepithelioma in 1 and inflammatory myofibroblastic tumor in 1. The median length of the resected bronchus was 25 mm (range 15-50 mm). Three patients (13%) had, at least, one postoperative complication with no deaths. Two patients had lymph node metastases and eight had positive margins. Nine patients received adjuvant therapy. Follow-up ranged from 13 to 96 months, all patients are currently alive and free of recurrence. Resection and reconstruction of the left secondary carina with preservation of the lung parenchyma can be performed safely in anatomically and oncologically appropriate patients, providing good short-term results when combined with adjuvant therapies.

Objectives A rare and complex procedure, total lung sparing left secondary carinal resection and reconstruction is only performed in a few specialized centers in a restricted group of patients. We reviewed our experience to evaluate its safety. Methods Patients who underwent left secondary carinal resection and reconstruction with complete lung parenchymal preservation for low-grade bronchial malignancies at the Shanghai Chest Hospital and the Padua University Hospital were retrospectively reviewed. Clinicopathologic factors and perioperative outcomes were analyzed. Results Thirty patients underwent the procedure between July 2012 and July 2019 (mean age, 42.9 years). No operative mortality occurred and postoperative complications developed in 4 patients (13.3%), including pneumonia (n = 3 [10.0%]), subcutaneous emphysema (n = 2 [6.7%]), and prolonged air leak (n = 2 [6.7%]). Pathologies included adenoid cystic carcinoma (n = 11), mucoepidermoid carcinoma (n = 6), carcinoid tumors (n = 9 [8 typical and 1 atypical subtypes]), inflammatory myofibroblastic tumor (n = 3), and myoepithelioma (n = 1). The margins were positive in 8 patients (26.7%), whereas 2 patients (6.7%) had positive lymph nodes. Adjuvant therapies were performed postoperatively, including chemoradiotherapy for positive lymph nodes and radiotherapy for positive margins. Conclusions Total lung sparing left secondary carinal resection and reconstruction can be performed safely in well-selected and oncologically appropriate patients with low-grade bronchial malignancies.

Sarcomeres are the force-generating and load-bearing devices of muscles. A precise molecular understanding of how the entire sarcomere is built is required to understand its role in health, disease and ageing. Here, we determine thein situmolecular architecture of vertebrate skeletal sarcomeres through electron cryo-tomography of cryo-focused ion beam-milled native myofibrils. The reconstructions reveal the three-dimensional organisation and interaction of actin and myosin filaments in the A-band, I-band and Z-disc and demonstrate how α -actinin cross-links antiparallel actin filaments to form a mesh-like structure in the Z-disc at an unprecedented level of molecular detail. A prominent feature is a so-far undescribed doublet of α-actinin cross-links with ∼ 6 nm spacing. Sub-volume averaging shows the interaction between myosin, tropomyosin and actin in molecular detail at ∼ 10 Å resolution and reveals two coexisting conformations of actin-bound heads. The flexible orientation of the lever arm and the essential and regulatory light chains allow the two heads of the “double-headed” myosin not only to interact with the same actin filament but also to split between two actin filaments. Our results provide new insights into the conformational plasticity and fundamental organisation of vertebrate skeletal muscle and serve as a strong foundation for futurein situinvestigations of muscle diseases.

Cryo-electron tomography (cryo-ET) is an emerging technique to study the cellular architecture and the structure of proteins at high resolution in situ . Most biological specimens are too thick to be directly investigated and are therefore thinned by milling with a focused ion beam under cryogenic conditions (cryo-FIB). This procedure is unautomated and prone to contaminations, which makes it a tedious and time-consuming process, often leading to suboptimal results. Here, we present new hardware and software tools that overcome the current limitations. We developed a new glove box and a high vacuum cryo transfer system and installed a stage heater, a cryo-shield and a cryo-shutter in the FIB milling microscope. This reduces the ice contamination during the transfer and milling process and simplifies the handling of the sample. In addition, we introduce a new software application that automates the key milling steps. Together, these improvements allow for high-quality, high-throughput cryo-FIB milling. This paves the way for new types of experiments, which have been previously considered infeasible.

BACKGROUND: Immune-related genes (IRGs) play an important role in the tumor immune microenvironment and affect tumor prognosis. This study aimed to establish a prognostic signature for malignant pleural mesothelioma (MPM) patients. METHODS: We obtained the relevant data of MPM patients in The Cancer Genome Atlas (TCGA), and univariate and multivariate Cox regression were used to construct the prediction signature and verify it with the external validation dataset GSE2549. A nomogram was then constructed, and its predictive ability was evaluated and analyzed the level of immune cell infiltration in different groups in the signature. RESULTS: An IRG-related prognostic signature composed of INHBA, CAT, SORT1, TNFSF13B, and BIRC5 was constructed, with patients divided into high-risk and low-risk groups according to the risk score. The survival time of overall survival (OS), progression-free survival (PFS), disease-free interval (DFI), and relapse-free survival (RFS) in low-risk groups was longer than in high-risk groups. Furthermore, the signature had high predictive performance, and the receiver operating characteristic (ROC) of 1, 2, and 3 years could reach 0.853, 0.881, and 0.914, respectively. The predictive accuracy of the signature was verified by using the independent GSE2549 dataset. The levels of activated CD4 T cells, immature dendritic cells, and type 2 T helper cells were higher in high-risk patients. The gene set enrichment analysis (GSEA) analysis showed that a high concentration and P53 signal pathways were found in high-risk groups. CONCLUSIONS: This research developed and verified a new type of immune prognostic signature based on five IRGs, which can predict the prognosis of tumor patients and provide new ideas for individualized treatment.

Septins are a family of eukaryotic guanosine phosphate-binding proteins that form linear heterooligomeric complexes, which, in turn, polymerize end-on-end into filaments. These filaments further assemble into higher-order structures at distinct subcellular locations. Dynamic changes in the organization of septin cortex structures appear during cell cycle progression. A variety of regulatory proteins and posttranslational modifications are involved in changes to the structure of septin assemblies during the entire cell cycle. In particular, septin-associated protein kinases mediate changes to septin higher order structures or interconnect cellular morphogenesis with the cell cycle. Yeast cyclin-dependent kinase, a master cell cycle regulator, is required for the initiation of a new septin ring. Here, using epifluoresence and electron microscopy, we show that upon phosphorylation by the Cdc28 kinase, septin filaments disassemble into hetero-octameric building blocks, and that filament depolymerization is specifically G1 cyclin-dependent.