ObjectiveTo evaluate the efficacy of combined rapid on-site evaluation of cytology (ROSE), ultrathin bronchoscopy, virtual bronchoscopic navigation, radial endobronchial ultrasound (EBUS), and metagenomic next-generation sequencing (mNGS) for diagnosis of peripheral pulmonary infectious lesions.MethodsSpecimens from patients with peripheral lung infection were obtained by transbronchial lung biopsy (TBLB) and bronchoalveolar lavage (BAL), and mNGS was used to detect pathogenic microorganisms. The sensitivity and specificity of mNGS were compared between TBLB tissue and BAL fluid.ResultsThe most common pathogens of pulmonary infectious lesions in this study were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii. The specificity of mNGS was higher in TBLB tissue than in BAL fluid, but mNGS of BAL fluid had higher sensitivity.ConclusionsThe combination of ROSE, ultrathin bronchoscopy, virtual bronchoscopic navigation, radial EBUS, and mNGS technology yielded high efficacy for the diagnosis of peripheral pulmonary infectious lesions. TBLB and BAL specimens have respective advantages in specificity and sensitivity for mNGS analysis.

Background: The purpose of this study was to evaluate the diagnostic value of combined virtual bronchoscopic navigation (Direct Path), radial endobronchial ultrasound with guide-sheath (EBUS), ultrathin bronchoscopy, rapid on-site evaluation of cytology (ROSE), and metagenomic next-generation sequencing (mNGS) for difficult lung lesions in patients with haematological diseases.Methods: In this study, lung specimens were obtained from patients with haematological diseases by transbronchial lung biopsy (TBLB) and bronchoalveolar lavage (BAL). The specimens were subjected to mNGS for sequencing of pathogenic microorganisms and sent to the laboratory for examination and pathological analysis. Additionally, the clinical data and pathogenic characteristics of the patients were analysed. The sensitivity and specificity of mNGS for sequencing pathogenic microorganisms were compared between TBLB and BAL specimens.Results: In this study, the diagnosis of infectious pneumonia mainly included cytomegalovirus pneumonia, Pneumocystis jirovecii pneumonia (PCP), pulmonary aspergillosis, and tuberculosis. Some patients had noninfectious pulmonary complications, and the clinical and therapeutic outcomes were diagnosed as graftversus-host disease (GVHD), idiopathic pneumonia syndrome (IPS), and delayed pulmonary toxicity syndrome (DPTS). The sensitivity of mNGS for pathogenic microbes in lung tissue is better than that of alveolar lavage fluid, whereas compared with alveolar lavage fluid, its specificity is reduced. Conclusions:The results of this study indicate that combined virtual bronchoscopic navigation (Direct Path), radial EBUS, ultrathin bronchoscopy, and ROSE of target control specimens reduce the risk of bleeding, and their combination with mNGS has high diagnostic value for difficult lung lesions in patients with haematological diseases, especially in the field of infection diagnosis. TBLB and BAL specimens have respective advantages in specificity and sensitivity for mNGS analysis.

Sepsis is a systemic inflammatory response syndrome caused by infections. The present study aimed to investigate the potential mechanism of FGD5-AS1 in sepsis and lipopolysaccharide (LPS)-induced inflammatory response. An animal model of sepsis was constructed. LPS was used to induce mice HL-1 cardiomyocytes to construct a cell model. The association between FGD5-AS1 and miR-133a-3p was investigated through animal and cell models. FGD5-AS1 overexpression was used to analyze the effect of FGD5-AS1 on inflammatory reaction. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 levels were detected by enzyme-linked immunosorbent assay and reverse transcription-quantitative polymerase chain reaction. The interaction of FGD5-AS1, miR-133a-3p and aquaporin 1 (AQP1) was detected by dual-luciferase reporter assay and microRNA (miRNA/miR) pull-down assay. Compared with the control group, the expression of FGD5-AS1 was decreased and the expression of miR-133a-3p was increased in the sepsis group. FGD5-AS1 overexpression increased LPS-induced expression of FGD5-AS1 and AQP1, decreased the expression of miR-133a-3p, and inhibited the expression of the inflammatory cytokines, TNF-α, IL-6 and IL-1β. Dual-luciferase reporter and miRNA pull-down assays confirmed the interaction of FGD5-AS1, miR-133a-3p and AQP1. These results indicated that FGD5-AS1 is the competitive endogenous RNA of miR-133a-3p on AQP1, and thus FGD5-AS1 overexpression may be able to inhibit the inflammatory response in sepsis.