PM2.5 induces a senescent state in mouse AT2 cells.

PM2.5 is known to induce lung injury, but its toxic effects on lung regenerative machinery and the underlying mechanisms remain unknown. In this study, primary mouse alveolar type 2 (AT2) cells, considered stem cells in the gas-exchange barrier, were sorted using fluorescence-activated cell sorting. By developing microfluidic technology with constricted microchannels, we observed that both passage time and impedance opacities of mouse AT2 cells were reduced after PM2.5, indicating that PM2.5 induced a more deformable mechanical property and a higher membrane permeability. In vitro organoid cultures of primary mouse AT2 cells indicated that PM2.5 is able to impair the proliferative potential and self-renewal capacity of AT2 cells but does not affect AT1 differentiation. Furthermore, cell senescence biomarkers, p53 and γ-H2A.X at protein levels, P16ink4a and P21 at mRNA levels were increased in primary mouse AT2 cells after PM2.5 stimulations as shown by immunofluorescent staining and quantitative PCR analysis. Using several advanced single-cell technologies, this study sheds light on new mechanisms of the cytotoxic effects of atmospheric fine particulate matter on lung stem cell behavior.

Knowledge mapping analysis of safety ergonomics: a bibliometric study.

Although a significant attention, the field of safety ergonomics has not yet been systematically profiled based on recent studies. To fully understand the current research status, basis, hotspots, and development trends in the field, 533 documents from the Web of Science core database were used for knowledge mapping analysis by the bibliometric method. The study found that the USA is the top country in publications, and Tehran University is the institution with the highest number of publications. Ergonomics and Applied Economics are the authoritative safety ergonomics journals. Through co-occurrence and co-citation analysis, current safety ergonomics research is focussed on healthcare, product design, and occupational health and safety. The keyword timeline view indicates that the main research paths are occupational health and safety, and patient safety research. The analysis of burst keywords shows that safety ergonomics research in management, model design, and system design areas are research frontiers in the field.Practitioner summary: This paper presents a knowledge mapping of safety ergonomics research through bibliometric analysis. The research results show the research status, research hotspots, and research frontiers in the field of safety ergonomics, which provides a direction for other scholars to quickly understand the development of this field.

Short-term PM2.5 exposure induces transient lung injury and repair.

Exposure to fine atmospheric particulate matter (PM) is known to induce lung inflammation and injury; however, the way in which sophisticated endogenous lung repair and regenerative programs respond to this exposure remains unknown. In this study, we established a whole-body mouse exposure model to mimic real scenarios. Exposure to fine PM (PM with an aerodynamic diameter ≤ 2.5 µm [PM2.5]; mean 1.05 mg/m3) for 1-month elicited inflammatory infiltration and epithelial alterations in the lung, which were resolved 6 months after cessation of exposure. Immune cells that responded to PM2.5 exposure mainly included macrophages and neutrophils. During PM2.5 exposure, alveolar epithelial type 2 cells initiated rapid repair of alveolar epithelial mucosa through proliferation. However, the reparative capacity of airway progenitor cells (club cells) was impaired, which may have been related to the oxidative production of neutrophils or macrophages, as suggested in organoid co-cultures. These data suggested that the pulmonary toxic effects of short-term exposure to fine atmospheric PM at a certain dosage could be overcome through tissue reparative mechanisms.

Nitric oxide hinders club cell proliferation through Gdpd2 during allergic airway inflammation.

Excessive nitric oxide (NO) is often observed in the airways of patients with severe asthma. Here, we show that the NO donor diethylamine NONOate impairs the proliferative capacity of mouse club cells and induces club cell apoptosis, cell cycle arrest, and alterations in lipid metabolism. Our data suggest that NO inhibits club cell proliferation via upregulation of Gdpd2 (glycerophosphodiester phosphodiesterase domain containing 2). During ovalbumin (OVA) challenge, apoptotic club cells are observed, but surviving club cells continue to proliferate. OVA exposure induces Gdpd2 expression; Gdpd2 knockout promotes the proliferation of club cells but inhibits goblet cell differentiation. Elimination of airway NO was found to inhibit goblet cell differentiation from club cells during OVA challenge. Our data reveal that excessive NO might be related to airway epithelial damage in severe asthma and suggest that blockade of the NO-Gdpd2 pathway may be beneficial for airway epithelial restoration.

Niche-Dependent Regulation of Lkb1 in the Proliferation of Lung Epithelial Progenitor Cells.

Lung homeostasis and regeneration depend on lung epithelial progenitor cells. Lkb1 (Liver Kinase B1) has known roles in the differentiation of airway epithelial cells during embryonic development. However, the effects of Lkb1 in adult lung epithelial progenitor cell regeneration and its mechanisms of action have not been determined. In this study, we investigated the mechanism by which Lkb1 regulates lung epithelial progenitor cell regeneration. Organoid culture showed that loss of Lkb1 significantly reduced the proliferation of club cells and alveolar type 2 (AT2) cells in vitro. In the absence of Lkb1, there is a slower recovery rate of the damaged airway epithelium in naphthalene-induced airway epithelial injury and impaired expression of surfactant protein C during bleomycin-induced alveolar epithelial damage. Moreover, the expression of autophagy-related genes was reduced in club cells and increased in AT2 cells, but the expression of Claudin-18 was obviously reduced in AT2 cells after Lkb1 knockdown. On the whole, our findings indicated that Lkb1 may promote the proliferation of lung epithelial progenitor cells via a niche-dependent pathway and is required for the repair of the damaged lung epithelium.

DJ-1 governs airway progenitor cell/eosinophil interactions to promote allergic inflammation.

BACKGROUND: DJ-1 is an antioxidant protein known to regulate mast cell-mediated allergic response, but its role in airway eosinophilic interactions and allergic inflammation is not known. OBJECTIVE: The aim of this study was to investigate the role of DJ-1 in airway eosinophilic inflammation in vitro and in vivo. METHODS: Ovalbumin-induced airway allergic inflammation was established in mice. ELISA was adopted to analyze DJ-1 and cytokine levels in mouse bronchoalveolar lavage fluid. Transcriptional profiling of mouse lung tissues was conducted by single-cell RNA-sequencing technology. The role of DJ-1 in the differentiation of airway progenitor cells into goblet cells was examined by organoid cultures, immunofluorescence staining, quantitative PCR, and cell transplantation in normal, DJ-1 knockout (KO), or conditional DJ-1 KO mice. RESULTS: This study observed that DJ-1 was increased in the lung tissues of ovalbumin-sensitized and challenged mice. DJ-1 KO mice exhibited reduced airway eosinophil infiltration and goblet cell differentiation. Mechanistically, we discovered that eosinophil-club cell interactions are reduced in the absence of DJ-1. Organoid cultures indicated that eosinophils impair the proliferative potential of club cells. Intratracheal transplantation of DJ-1-deficient eosinophils suppresses airway goblet cell differentiation. Loss of DJ-1 inhibits the metabolism of arachidonic acid into cysteinyl leukotrienes in eosinophils while these secreted metabolites promote airway goblet cell fate in organoid cultures and in vivo. CONCLUSIONS: DJ-1-mediated interactions between airway epithelial progenitor cells and immune cells are essential in controlling airway goblet cell metaplasia and eosinophilia. Blockade of the DJ-1 pathway is protective against airway allergic inflammation.

Glutamine Metabolism Is Required for Alveolar Regeneration during Lung Injury.

(1) Background: Abnormal repair after alveolar epithelial injury drives the progression of idiopathic pulmonary fibrosis (IPF). The maintenance of epithelial integrity is based on the self-renewal and differentiation of alveolar type 2 (AT2) cells, which require sufficient energy. However, the role of glutamine metabolism in the maintenance of the alveolar epithelium remains unclear. In this study, we investigated the role of glutamine metabolism in AT2 cells of patients with IPF and in mice with bleomycin-induced fibrosis. (2) Methods: Single-cell RNA sequencing (scRNA-seq), transcriptome, and metabolomics analyses were conducted to investigate the changes in the glutamine metabolic pathway during pulmonary fibrosis. Metabolic inhibitors were used to stimulate AT2 cells to block glutamine metabolism. Regeneration of AT2 cells was detected using bleomycin-induced mouse lung fibrosis and organoid models. (3) Results: Single-cell analysis showed that the expression levels of catalytic enzymes responsible for glutamine catabolism were downregulated (p < 0.001) in AT2 cells of patients with IPF, suggesting the accumulation of unusable glutamine. Combined analysis of the transcriptome (p < 0.05) and metabolome (p < 0.001) revealed similar changes in glutamine metabolism in bleomycin-induced pulmonary fibrosis in mice. Mechanistically, inhibition of the key enzymes involved in glucose metabolism, glutaminase-1 (GLS1) and glutamic-pyruvate transaminase-2 (GPT2) leads to reduced proliferation (p < 0.01) and differentiation (p < 0.01) of AT2 cells. (4) Conclusions: Glutamine metabolism is required for alveolar epithelial regeneration during lung injury.

Organoid technology and lung injury mouse models evaluating effects of hydroxychloroquine on lung epithelial regeneration.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) damages lung epithelial stem/progenitor cells. Ideal anti-SARS-CoV-2 drug candidates should be screened to prevent secondary injury to the lungs. Here, we propose that in vitro three-dimensional organoid and lung injury repair mouse models are powerful models for the screening antiviral drugs. Lung epithelial progenitor cells, including airway club cells and alveolar type 2 (AT2) cells, were co-cultured with supportive fibroblast cells in transwell inserts. The organoid model was used to evaluate the possible effects of hydroxychloroquine, which is administered as a symptomatic therapy to the coronavirus disease 2019 (COVID-19) patients, on the function of mouse lung stem/progenitor cells. Hydroxychloroquine was observed to promote the self-renewal of club cells and differentiation of ciliated and goblet cells in vitro. Additionally, it inhibited the self-renewal ability of AT2 cells in vitro. Naphthalene- or bleomycin-induced lung injury repair mouse models were used to investigate the in vivo effects of hydroxychloroquine on the regeneration of club and AT2 cells, respectively. The naphthalene model indicated that the proliferative ability and differentiation potential of club cells were unaffected in the presence of hydroxychloroquine. The bleomycin model suggested that hydroxychloroquine had a limited effect on the proliferation and differentiation abilities of AT2 cells. These findings suggest that hydroxychloroquine has limited effects on the regenerative ability of epithelial stem/progenitor cells. Thus, stem/progenitor cell-derived organoid technology and lung epithelial injury repair mouse models provide a powerful platform for drug screening, which could possibly help end the pandemic.

LKB1 deficiency upregulates RELM-α to drive airway goblet cell metaplasia.

Targeting airway goblet cell metaplasia is a novel strategy that can potentially reduce the chronic obstructive pulmonary disease (COPD) symptoms. Tumor suppressor liver kinase B1 (LKB1) is an important regulator of the proliferation and differentiation of stem/progenitor cells. In this study, we report that LKB1 expression was downregulated in the lungs of patients with COPD and in those of cigarette smoke-exposed mice. Nkx2.1Cre; Lkb1f/f mice with conditional loss of Lkb1 in mouse lung epithelium displayed airway mucus hypersecretion and pulmonary macrophage infiltration. Single-cell transcriptomic analysis of the lung tissues from Nkx2.1Cre; Lkb1f/f mice further revealed that airway goblet cell differentiation was altered in the absence of LKB1. An organoid culture study demonstrated that Lkb1 deficiency in mouse airway (club) progenitor cells promoted the expression of FIZZ1/RELM-α, which drove airway goblet cell differentiation and pulmonary macrophage recruitment. Additionally, monocyte-derived macrophages in the lungs of Nkx2.1Cre; Lkb1f/f mice exhibited an alternatively activated M2 phenotype, while expressing RELM-α, which subsequently aggravated airway goblet cell metaplasia. Our findings suggest that the LKB1-mediated crosstalk between airway progenitor cells and macrophages regulates airway goblet cell metaplasia. Moreover, our data suggest that LKB1 agonists might serve as a potential therapeutic option to treat respiratory disorders associated with goblet cell metaplasia.

Enhancing higher-order eigenmodes of AFM using bridge/cantilever coupled system.

The wide application of multi-frequency atomic force microscopy (AFM) places higher demands on the higher-order modes response of the cantilever. The response of the higher modes however is generally weaker than that of the fundamental mode in air. Researchers have proposed many methods, most of which involve cantilever modification, to enhance higher-order eigenmodes response. These previous results are proved to be effective, but the microfabrication is expensive. In this article, we propose a novel model based on bridge/cantilever coupled system to enhance the higher-order modes response of AFM cantilever. The segmented beam model provides a new thinking to explain the appearance of undesired peaks in mode analysis of cantilever. Through theoretical analysis and simulation, we find that higher resonance modes are enhanced by tuning the bridge to match the high resonances of the single clamped cantilever. The length, thickness of the coupled system and the location of excitation can affect the enhancement. In summary, this model provides a new way to improve higher mode response for multi-frequency and other high bandwidth applications of AFM.

Reducing molecular simulation time for AFM images based on super-resolution methods.

Atomic force microscopy (AFM) has been an important tool for nanoscale imaging and characterization with atomic and subatomic resolution. Theoretical investigations are getting highly important for the interpretation of AFM images. Researchers have used molecular simulation to examine the AFM imaging mechanism. With a recent flurry of researches applying machine learning to AFM, AFM images obtained from molecular simulation have also been used as training data. However, the simulation is incredibly time consuming. In this paper, we apply super-resolution methods, including compressed sensing and deep learning methods, to reconstruct simulated images and to reduce simulation time. Several molecular simulation energy maps under different conditions are presented to demonstrate the performance of reconstruction algorithms. Through the analysis of reconstructed results, we find that both presented algorithms could complete the reconstruction with good quality and greatly reduce simulation time. Moreover, the super-resolution methods can be used to speed up the generation of training data and vary simulation resolution for AFM machine learning.

An Interface Optimization Strategy for g-C3N4-Based S-Scheme Heterojunction Photocatalysts.

Graphitic carbon nitride (CN) has attracted much attention in photocatalytic fields due to its unique electronic band structure. However, the rapid recombination of photogenerated carriers severely inhibits its catalytic activity. The heterojunction structure has been widely confirmed to significantly improve the photocatalytic activity of CN through the formed interface structure. However, researchers often give attention to the band matching and conductivity of the cocatalyst, while the importance of the interface as a migration channel for photogenerated carriers is often overlooked. In this work, we adopt the strategy of morphology engineering to regulate the morphology of the CN photoactive component so as to achieve the interface optimization of the traditional heterojunction structure. The photocatalytic degradation experiment of rhodamine B shows that compared with the traditional CeO2@CN heterojunction structure, the photocatalytic activity of the interface-optimized CeO2/CN is increased by more than 20%. The following points could be used to explain the improvement of photocatalytic activity: (I) the formed S-scheme heterojunction structure, which inhibits the recombination of useful electrons and holes but expedites the recombination of relatively useless electrons and holes, (II) the increased interface area, which provides more carrier migration channels, and (III) the reduced interface contact resistance, which facilitates the separation and migration of photogenerated carriers. Furthermore, the interface optimization of the traditional Al2O3@CN and Fe2O3@CN heterojunction structures also achieved consistent results. This shows that the strategy in this work is a universal method for interface optimization, which provides potential alternative for further improving the catalytic activity of other heterojunction composites.

Inhibition of Gabrp reduces the differentiation of airway epithelial progenitor cells into goblet cells.

Bronchial asthma is an intractable pulmonary disease that affects millions of individuals worldwide, with the overproduction of mucus contributing to high morbidity and mortality. Gamma-aminobutyric acid (GABA) is associated with goblet cell hyperplasia in the lungs of primate models and Club cells serve as airway epithelial progenitor cells that may differentiate into goblet and ciliated cells. In the present study, it was investigated whether the GABAA receptor pi (Gabrp) is essential for Club cell proliferation and differentiation in mice. Validation of microarray analysis results by reverse transcription-quantitative PCR (RT-qPCR) demonstrated that Gabrp is highly expressed in mouse Club cells. Predominant expression of Gabrp in mouse Club cells was further confirmed based on naphthalene-induced Club cell injury in mice, with organoid cultures indicating significant reductions in the organoid-forming ability of mouse Club cells in the presence of Gabrp antagonist bicuculline methiodide (BMI). Furthermore, the RT-qPCR results indicated that the mRNA levels of chloride channel accessory 3, pseudogene (Clca3p), mucin (Muc)5Ac and Muc5B were significantly decreased in BMI organoid cultures. These results suggested that blocking GABA signaling through Gabrp inhibits mouse Club cell proliferation, as well as differentiation into goblet cells. Therefore, targeting GABA/Gabrp signaling may represent a promising strategy for treating goblet cell hyperplasia in bronchial asthma.

Organoid models in lung regeneration and cancer.

Improper regeneration is associated with lung diseases including lung cancer. Lung cancer is one of the leading causes of death worldwide, with nearly 2 million new cases diagnosed each year. The diagnosis is often too late for successful therapeutic intervention. Lung cancer shows substantial phenotypic and genetic heterogeneity between individuals, making it difficult to model in animals. Organoids, derived from regional stem/progenitor cells in lung epithelia, have attracted extensive interest in both research studies and the clinic, because of their great potential for use in cancer treatment. Various lung cancer organoids have been established to recapitulate the tissue architecture of primary lung tumors and maintain the genomic alterations of the original tumors during long-term expansion in vitro. In this review, we summarize the current data on lung epithelial regeneration by regional endogenous stem/progenitor cells, describe the development of organoid technology, and present its applications in lung cancer research. Furthermore, recent challenges and future directions to improve organoid technologies for lung cancer treatment are discussed.

Biaxial fatigue crack propagation behavior of ultrahigh molecular weight polyethylene reinforced by carbon nanofibers and hydroxyapatite.

Ultrahigh molecular weight polyethylene (UHMWPE) artificial joint has remained the preferred polymer component in total joint replacement surgery. However, more and more concerns have been raised about the failure of UHMWPE components due to the initiation and propagation of cracks at the notches with fixed functions. For this reason, biaxial fatigue crack growth (FCG) experiments of UHMWPE reinforced by carbon nanofibers (CNF) and hydroxyapatite (HA) were carried out using elastic-plastic fracture mechanics theory. The FCG resistance of UHMWPE, UHMWPE/CNF, and UHMWPE/HA was compared, and the effects of stress ratio (R) value and phase difference on FCG rate were investigated. At the same time, the influence of loading path was considered, and the corresponding crack path was analyzed. Results suggest that UHMWPE/CNF has better FCG resistance and the FCG rate increases with the increase of R value and the existence of 180° phase difference. In addition, crack bifurcation behavior is not observed under nonproportional loading conditions. The findings in this study will provide experimental validation and data support for better clinical application of UHMWPE-modified materials.

A novel transgenic murine model with persistently brittle bones simulating osteogenesis imperfecta type I.

Osteogenesis imperfecta (OI) type I caused by the null allele of COL1A1 gene is in the majority in clinical OI cases. Currently, heterozygous Mov-13 mice generated by virus insertion in the first intron of col1a1 is the exclusive model to modulate OI type I, in spite of the gradually recovered bone mineral and mechanical properties. A newly designed heterozygous col1a1±365 OI mouse was produced in the present study by partial exons knockout (exon 2-exon 5, 365 nt of mRNA) using CRISPR/Cas9 system. The deletion resulted in generally large decrease in type I collagen synthesis due to frameshift mutation and premature chain termination, closely mimicking the pathogenic mechanism in affected individuals. And the strain possessed significantly sparse mineral scaffolds, bone loss, lowered mechanical strength and broken bone metabolism by 8 and 20 weeks compared to their littermates, suggesting a sustained skeletal weakness. Notably, the remarkable down-regulation of Yes-associated protein (YAP), one of the key coactivator in Hippo signaling pathway, was first found both in the femur and adipose derived mesenchymal stem cells (ADSCs) under osteogenic differentiation of col1a1±365 mice, which might be responsible for the reduced osteogenic potential and brittle bones. Still, further research was needed in order to illuminate the underlying mechanism.

Carbon Nitride Co-catalyst Activation Using N-Doped Carbon with Enhanced Photocatalytic H2 Evolution.

Photocatalytic water splitting holds huge potential to meet the current challenges of energy and environments. Among them, polymeric carbon nitride (CN) has drawn much attention as a promising metal-free photocatalyst. As it is known, a number of promising co-catalysts have been developed to improve catalytic reactions, Pt nanoparticles is still among the best co-catalysts for CN in photocatalytic H2 evolution, due to the suitable Fermi level to transfer excited electrons and the low overpotential for H2 reduction. Herein, we report the interface engineering of urea-derived bulk CN and Pt co-catalyst by using a small portion of N-doped carbon (N-C) as a transition layer with a boosted photocatalytic activity up to 7 times. It was revealed that the activation energy of the Pt co-catalyst for water reduction was lowered in the presence of N-C, and the intimate interaction between CN and N-C, ascribing to the similar elemental composition and crystal structure, promoted the efficient separation and migration of charge carriers. This study may open a new avenue to develop CN-based photocatalysts for solar fuel conversion with even higher activity by photocatalyst/co-catalyst interface engineering.

iTRAQ-based quantitative proteomic analysis reveals potential early diagnostic markers in serum of acute cellular rejection after liver transplantation.

Liver transplantation (LT) is the most effective treatment method for advanced stage liver disease but acute cellular rejection (ACR) seriously affects the prognosis of LT. To discover novel diagnostic biomarkers of ACR after LT, Isobaric Tags for Relative and Absolute Quantitation (iTRAQ)-based mass spectrometry was performed to characterize alterations of serum proteins among patients validated to be pathologically ACR or pathologically no-ACR after LT and healthy controls. As a result, 10 differentially expressed proteins were found out between the ACR group and the No-ACR group; 88 differentially expressed proteins were found out between the ACR group and the Healthy Control group; 39 differentially expressed proteins were found out between No-ACR group and Healthy Control group. After analysis and ELISA validation, the results showed that CFHR1, CFHR5 and CFH could be candidate protein biomarkers for the early diagnosis of ACR after LT.

Ratcheting behavior of UHMWPE reinforced by carbon nanofibers (CNF) and hydroxyapatite (HA): Experiment and simulation.

Uniaxial tensile tests were performed to investigate the mechanical properties of the ultra-high molecular weight polyethylene (UHMWPE) with different modification conditions. It was found that the different modification conditions have great influence on the mechanical properties of the UHMWPE. Subsequently, the uniaxial ratcheting behaviors of the UHMWPE/CNF and UHMWPE/HA composite materials were observed under the stress-controlled cyclic tensile condition at room temperature. The dependence of uniaxial ratcheting of composite materials on the mean stress, stress amplitude, stress rate and nano-material content was investigated. The results show that the ratcheting strain and its rate of the two composite materials increase as the mean stress and stress amplitude increase, however, the ratcheting strain and its rate decrease with the increase of the stress rate and nano-material content. Furthermore, it is found that the ratcheting strain of the UHMWPE/HA composite material is more remarkable than that of the UHMWPE/CNF composite material. A new viscoplastic constitutive model is proposed to describe the ratcheting behavior of the UHMWPE composite materials. In this model, a new viscosity function and modified kinematic hardening law were employed. Comparison of simulation and experimental results shows that the simulations are in good agreement with the experimental results.