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Mechanosensing and mechanotransduction are important processes that regulate pulmonary motility. Piezo2, as a mechanosensitive ion channel, was reported to be involved in the regulation of respiratory function in mice. However, it is unclear whether Piezo2 expression plays a role in disease progression in pulmonary dysfunction patients (such as ARDS patients). The aim of this study was to compare the expression of Piezo2 in the bronchoalveolar lavage fluid (BALF) of ARDS patients and the association between Piezo2 and the prognosis of ARDS patients. This study was approved by the Biomedical Ethics Review Committee of West China Hospital of Sichuan University. Patients diagnosed with ARDS from May 2022 to December 2022 in the Department of Critical Care Medicine, West China Hospital, Sichuan University were prospectively included. Then BALF was collected from every patient. The BALF was centrifuged, and the supernatant was taken for ELISA analysis. Basic information of the patients was recorded through the electronic medical record information system. We finally included 80 patients in this study. Compared with mild ARDS group, the level of piezo2 in BALF was significantly higher in moderate ARDS group (54.11 ± 8.113, 95%CI 37.75–70.48, P < 0.0001) and severe ARDS group (84.10 ± 15.44, 95%CI 53–115.2, P < 0.0001). To assess the Piezo2 value in predicting the development of ARDS patients, we graphed the ROC curve. It is revealed that Piezo2 in BALF could distinguish ARDS patients of different severity with areas under the curve (AUC) 0.9358 (95%CI 0.8667–1.00, P < 0.0001). Patients with higher level of Piezo2 had higher APACHE II score (16 ± 6.517 vs 24.4 ± 6.845, 95%CI 4.983–11.81, P < 0.0001), longer length of ICU stays (16.71 ± 9.717 vs 28.21 ± 13.78, 95%CI 4.84–18.15, P = 0.001). There was no significant difference in mortality with two groups (OR 1.42, 95%CI 0.43–4.66, P = 0.57) Higher level of Piezo2 was correlated to worse progression of ARDS patients.

The purpose of this study was to assess the value of metagenomic next-generation sequencing (mNGS) of bronchoalveolar lavage fluid (BALF) for the diagnosis of severe respiratory diseases based on interpretation of sequencing results. BALF samples were harvested and used for mNGS as well as microbiological detection. Infectious bacteria or fungi were defined according to relative abundance and number of unique reads. We performed mNGS on 35 BALF samples from 32 patients. The positive rate reached 100% in the mNGS analysis of nine immunocompromised patients. Compared with the culture method, mNGS had a diagnostic sensitivity of 88.89% and a specificity of 74.07% with an agreement rate of 77.78% between these two methods. Compared with the smear method and PCR, mNGS had a diagnostic sensitivity of 77.78% and a specificity of 70.00%. In 13 cases, detection results were positive by mNGS but negative by culture/smear and PCR. The mNGS findings in 11/32 (34.4%) cases led to changes in treatment strategies. Linear regression analysis showed that diversity was significantly correlated with interval between disease onset and sampling. Dynamic changes in reads could indirectly reflect therapeutic effectiveness. BALF mNGS improves sensitivity of pathogen detection and provides guidance in clinical practice. Potential pathogens can be identified based on relative abundance and number of unique reads.

Metagenomic next-generation sequencing (mNGS) is a powerful method for pathogen detection. In this study, we assessed the value of mNGS for bronchoalveolar lavage (BAL) samples in the diagnosis of pulmonary infections. From February 2018 to April 2019, BAL samples were collected from 235 patients with suspected pulmonary infections. mNGS and microbial culture were performed to evaluate the effectiveness of mNGS in pulmonary infection diagnosis. We employed mNGS to evaluate the alpha diversity, results suggesting that patients with confirmed pathogens had a lower microbial diversity index compared to that of patients with uncertain pathogens. For the patients admitted to the respiratory intensive care unit (RICU) or on a ventilator, they experienced a lower diversity index than that of the patients in the general ward or not on a ventilator. In addition, mNGS of BAL had a diagnostic sensitivity of 88.89% and a specificity of 14.86% in pulmonary infection, with 21.16% positive predictive value (PPV) and 83.87% negative predictive value (NPV). When rare pathogens were excluded, the sensitivity of mNGS decreased to 73.33%, and the specificity increased to 41.71%. For patients in the simple pulmonary infection group and the immunocompromised group, the main infection types were bacterial infection (58.33%) and mixed-infection (43.18%). Furthermore, mNGS had an advantage over culture in describing polymicrobial ecosystem, demonstrating the microbial distribution and the dominant strains of the respiratory tract in patients with different underlying diseases. The study indicated that mNGS of BAL samples could provide more accurate diagnostic information in pulmonary infections and demonstrate the changes of respiratory microbiome in different underlying diseases. This method might play an important role in the clinical use of antimicrobial agents in the future.

Extracellular vesicles (EVs) are cell-derived membranous vesicles secreted by cells into the extracellular space, which play a role in cell to cell communication. EVs are categorized into 3 groups depending on their size, surface marker, and method of release from the host cell. Recently, EVs have become of interest in the study of multiple disease etiologies and are believed to be potential biomarkers for many diseases. Multiple different methods have been developed to isolate EVs from different samples such as cell culture medium, serum, blood, and urine. Once isolated, EVs can be characterized by technology such as nanotracking analysis, dynamic light scattering, and nanoscale flow cytometry. In this review, we summarize the current methods of EV isolation, provide details into the three methods of EV characterization, and provide insight into which isolation approaches are most suitable for EV isolation from bronchoalveolar lavage fluid (BALF).

Background The reported incidence of aspergillosis among COVID-19 patients has varied significantly, which can be partly attributed to differences in diagnostic approaches and levels of physicians’ proficiency in diagnosing COVID-19-associated pulmonary aspergillosis (CAPA). Consequently, we conducted a retrospective study to investigate the potential reasons for these discrepancies and analyzed the risk factors for pulmonary aspergillosis in patients with COVID-19. Method Data were retrospectively collected from December 1, 2022, to September 30, 2023, from patients who were admitted to the First Affiliated Hospital of Wenzhou Medical University. The research platform was used to screen patients with discharge diagnoses of COVID-19 pneumonia. CAPA was defined according to the 2020 ECMM/ISHAM criteria and the Chinese expert consensus. Clinical data that were collected included data about underlying diseases, laboratory examinations and microbiological detection. Analyses were conducted with R software, with continuous variables analyzed with t-tests, categorical variables analyzed with chi-square tests, and logistic regression and ROC curves used to assess risk factors for CAPA. Results The incidence of CAPA was 13.4% in the general ward, 30.8% in the RICU, and 6.8% in other ICUs. The average time to CAPA diagnosis was 5.6 days in general wards, 3.7 days in the RICU, and 7.4 days in other ICUs. Diagnostic testing revealed the following sensitivities: 78% for BALF galactomannan (GM), 48% for serum GM, 52% for culture tests, and 71% for BALF mNGS. Risk factors for CAPA included chronic respiratory disease, chronic renal insufficiency, and diabetes. The primary Aspergillus species identified was A. fumigatus , followed by A. flavus . Conclusion Differences in incidence may arise from varying levels of physician awareness, which can influence the rate at which BALF and serum GM samples are submitted for testing. The sensitivity of BALF GM is higher than that of serum GM. Furthermore, BALF mNGS has the potential to enhance the clinical detection sensitivity of CAPA. Risk factors for CAPA include chronic respiratory disease, chronic renal insufficiency, and diabetes, which may aid in identifying at-risk patients. The primary Aspergillus species identified was A. fumigatus , followed by A. flavus , providing a reference for clinical empirical treatment. Clinical trial number Not applicable.

Chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF) are chronic, progressive lung ailments that are characterized by distinct pathologies. Early detection biomarkers and disease mechanisms for these debilitating diseases are lacking. Extracellular vesicles (EVs), including exosomes, are small, lipid-bound vesicles attributed to carry proteins, lipids, and RNA molecules to facilitate cell-to-cell communication under normal and diseased conditions. Exosomal miRNAs have been studied in relation to many diseases. However, there is little to no knowledge regarding the miRNA population of bronchoalveolar lavage fluid (BALF) or the lung-tissue-derived exosomes in COPD and IPF. Here, we determined and compared the miRNA profiles of BALF- and lung-tissue-derived exosomes of healthy non-smokers, smokers, and patients with COPD or IPF in independent cohorts. Results: Exosome characterization using NanoSight particle tracking and TEM demonstrated that the BALF-derived exosomes were ~89.85 nm in size with a yield of ~2.95 × 1010 particles/mL in concentration. Lung-derived exosomes were larger in size (~146.04 nm) with a higher yield of ~2.38 × 1011 particles/mL. NGS results identified three differentially expressed miRNAs in the BALF, while there was one in the lung-derived exosomes from COPD patients as compared to healthy non-smokers. Of these, miR-122-5p was three- or five-fold downregulated among the lung-tissue-derived exosomes of COPD patients as compared to healthy non-smokers and smokers, respectively. Interestingly, there were a large number (55) of differentially expressed miRNAs in the lung-tissue-derived exosomes of IPF patients compared to non-smoking controls. Conclusions: Overall, we identified lung-specific miRNAs associated with chronic lung diseases that can serve as potential biomarkers or therapeutic targets.

In the longtime challenge of identifying specific, easily detectable and reliable biomarkers of IPF, BALF proteomics is providing interesting new insights into its pathogenesis. To the best of our knowledge, the present study is the first shotgun proteomic investigation of EVs isolated from BALF of IPF patients. Our main aim was to characterize the proteome of the vesicular component of BALF and to explore its individual impact on the pathogenesis of IPF. To this purpose, ultracentrifugation was chosen as the EVs isolation technique, and their purification was assessed by TEM, 2DE and LC-MS/MS. Our 2DE data and scatter plots showed considerable differences between the proteome of EVs and that of whole BALF and of its fluid component. Analysis of protein content and protein functions evidenced that EV proteins are predominantly involved in cytoskeleton remodeling, adenosine signaling, adrenergic signaling, C-peptide signaling and lipid metabolism. Our findings may suggest a wider system involvement in the disease pathogenesis and support the importance of pre-fractioning of complex samples, such as BALF, in order to let low-abundant proteins-mediated pathways emerge.

Pneumonia caused by Streptococcus pneumoniae remains a serious threat to the elderly due to weakened immunity and limited treatment efficacy. This study investigated the antibacterial activity and therapeutic potential of Bletilla striata ethanolic extract (BSE) in a rat model of pneumococcal pneumonia. BSE demonstrated selective antibacterial activity against S. pneumoniae (MIC: 1.25 mg/mL) and significantly inhibited Sortase A (SrtA), a key bacterial virulence factor. In vivo, BSE improved body weight, reduced lung wet/dry ratios, and lowered serum ALP and LDH levels. It also diminished inflammatory cell infiltration in bronchoalveolar lavage fluid and decreased TNF-α, IL-1β, and IL-6 levels. Antioxidant markers SOD and GSH increased, while MDA levels declined. Histological analysis showed preserved lung architecture with reduced edema and cellular infiltration. Mechanistically, BSE suppressed the TLR4/NF-κB pathway. These findings indicate that BSE can offer a dual benefit—direct antimicrobial action and modulation of inflammation and oxidative stress—highlighting its potential as a natural therapeutic for elderly patients with pneumonia.

Background: Idiopathic pulmonary fibrosis (IPF) is a rare and severe disease with a median survival of ∼3 years. Several risk factors have been identified, such as age, genetic predisposition, tobacco exposure, and gastro-oesophageal reflux disease (GERD). Prevalence of GERD in IPF is high and may affect 87% of patients, of whom only half (47%) report symptoms. Objective: The aim of this study is to review current evidence regarding the correlation between GERD and IPF and to evaluate the current studies regarding treatments for GERD-IPF. Methods: A review to identify research papers documenting an association between GERD and IPF was performed. Results: We identified several studies that have confirmed the association between GERD and IPF, with an increased acid exposure, risk of gastric aspiration and bile acids levels in these patients. Few studies focused their attention on GERD treatment, showing how antiacid therapy was not able to change IPF evolution. Conclusions: This review investigating the correlation between GERD and IPF has confirmed the hypothesized association. However, further large prospective studies are needed to corroborate and elucidate these findings with a focus on preventative and treatment strategies.

[This corrects the article DOI: 10.3389/fmed.2025.1660190.].