Bronchoalveolar Lavage Fluid Of Patients Research Articles

Single-cell landscape of bronchoalveolar immune cells in patients with immune checkpoint inhibitor-related pneumonitis

The pathophysiology of immune checkpoint inhibitor-related pneumonitis remains incompletely understood. We conducted single-cell and T-cell receptor transcriptomic sequencing on the bronchoalveolar lavage fluid from five patients with grade ≥2 immune checkpoint inhibitor-related pneumonitis. Our analyses revealed a prominent enrichment of T cells in the bronchoalveolar lavage fluid of patients with immune checkpoint inhibitor-related pneumonitis. Within the CD4 + T cell subset, Tfh-like T cells were highly enriched and exhibited signatures associated with inflammation and clonal expansion. Regulatory T cells were also enriched and displayed enhanced inhibitory functions. Within the CD8 + T-cell subset, effector memory/tissue-resident memory T cells with an elevated cytotoxic phenotype were highly infiltrated. Among myeloid cells, alveolar macrophages were depleted, while pro-inflammatory intermediate monocytes were elevated. Dendritic cells demonstrated enhanced antigen presentation capabilities. Cytokines CXCR4, CXCL13, TNF-α, IFN-α, IFN-γ, and TWEAK were elevated. Through a comprehensive single-cell analysis, we depicted the landscape of immune checkpoint inhibitor-related pneumonitis.

Identification and assessment of antifungal susceptibility of Candida species based on bronchoalveolar lavage in immunocompromised and critically ill patients

Background and Objectives: The presence of fungi in the respiratory tract as mycobiome, particularly Candida species (spp.), remains a serious problem due to increasing numbers of immunocompromised patients. The confirmed reliable ex- istence of these pathogens due to frequent colonization is essential. This investigation aimed to recognize Candida spp. among isolates from bronchoalveolar lavage of immunocompromised and critically ill patients and to evaluate their suscep- tibility to antimycotic drugs. Materials and Methods: Bronchoalveolar lavage fluid was collected from 161 hospitalized patients presenting with sus- pected respiratory fungal infection /colonization. The specimens were examined by standard molecular and mycological assays. Candida spp. were recognized with sequence assessment of the D1-D2 section of the large subunit ribosomal DNA. The susceptibility of Candida isolates to common antimycotic drugs was distinguished by standard broth microdilution. Results: Seventy-one clinical isolates of Candida spp. were recognized. Candida albicans was the most frequent, followed by C. glabrata, C. krusei (Pichia kudriavzevii), C. dubliniensis, C. parapsilosis, and C. tropicalis. We found 5.1% of C. albi- cans isolates and 8% of C. glabrata isolates to show resistance to fluconazole. The whole of the Candida spp. were sensitive to amphotericin B and caspofungin. Conclusion: This study demonstrated that C. albicans and C. glabrata are the most common isolates of bronchoalveolar lavage fluid in patients, and the drug susceptibility screening confirmed that amphotericin B and caspofungin are effective against Candida spp. but some C. glabrata and C. albicans isolates showed resistance to fluconazole.

Microbiome features in bronchoalveolar lavage fluid of patients with idiopathic inflammatory myopathy-related interstitial lung disease.

Interstitial lung disease (ILD) is a common complication of idiopathic inflammatory myopathy (IIM), which is one of the connective tissue diseases (CTD). It can lead to poor prognosis and increased mortality. However, the distribution and role of the lower respiratory tract (LRT) microbiome in patients with IIM-ILD remains unclear. This study aimed to investigate the microbial diversity and community differences in bronchoalveolar lavage fluid (BALF) in patients with IIM-ILD. From 28 June 2021 to 26 December 2023, 51 individual BALF samples were enrolled, consisting of 20 patients with IIM-ILD, 16 patients with other CTD-ILD (including 8 patients with SLE and 8 with RA) and 15 patients with CAP. The structure and function of microbiota in BALF were identified by metagenomic next-generation sequencing (mNGS). The community evenness of LRT microbiota within the IIM-ILD group was marginally lower compared to the other CTD-ILD and CAP groups. Nonetheless, there were no noticeable differences. The species community structure was similar among the three groups, based on the Bray-Curtis distance between the samples. At the level of genus, the IIM-ILD group displayed a considerably higher abundance of Pseudomonas and Corynebacterium in comparison to the CAP group (p < 0.01, p < 0.05). At the species level, we found that the relative abundance of Pseudomonas aeruginosa increased significantly in the IIM-ILD group compared to the CAP group (p < 0.05). Additionally, the relative abundance of Prevotella pallens was significantly higher in other CTD-ILD groups compared to that in the IIM-ILD group (p < 0.05). Of all the clinical indicators examined in the correlation analysis, ferritin level demonstrated the strongest association with LRT flora, followed by Serum interleukin-6 level (p < 0.05). Our research has identified particular LRT microorganisms that were found to be altered in the IIM-ILD group and were significantly associated with immune function and inflammatory markers in patients. The lower respiratory tract microbiota has potential in the diagnosis and treatment of IIM-ILD.

Comparison of the adherence of nontypeable haemophilus influenzae to lung epithelial cells

ObjectiveNontypeable Haemophilus influenzae (NTHi) plays an important role in respiratory tract infections, and adherence to lung epithelial cells is the first step in lung infections. To explore the role of NTHi in childhood lung infections, a comparative study was conducted on the adherence of strains isolated from sputum culture and bronchoalveolar lavage fluid to A549 lung epithelial cells.MethodsHaemophilus influenzae strains were obtained from the sample bank of Shenzhen Children’s Hospital, and identified as NTHi via PCR detection of the capsule gene bexA. NTHi obtained from healthy children’s nasopharyngeal swabs culture were selected as the control group, and a comparative study was conducted on the adherence of strains isolated from sputum culture or bronchoalveolar lavage fluid of patients to A549 cells.ResultsThe adherence bacterial counts of NTHi isolated from the nasopharyngeal cultures of healthy children to A549 cells was 58.2 CFU. In patients with lung diseases, NTHi isolated from bronchoalveolar lavage fluid was 104.3 CFU, and from sputum cultures was 115.1 CFU, both of which were significantly higher in their adherence to A549 cells compared to the strains isolated from the healthy control group. There was no significant difference in adherence between the strains isolated from sputum cultures and bronchoalveolar lavage fluid (t = 0.5217, p = 0.6033).ConclusionNTHi played an important role in childhood pulmonary infections by enhancing its adherence to lung epithelial cells.

A Different Perspective on the COVID-19 Pandemic: Origin of the Outbreak (Part 1).

In December 2019, an outbreak caused by novel coronavirus 2019 (COVID-19) started in Wuhan, China. After extensive speculation about a causative agent, a novel betacoronavirus, in the same family as SARS-CoV and MERS-CoV, was identified via next-generation sequencing from samples received from several pneumonia patients. Ribonucleic acid extracted from the patient's bronchoalveolar lavage fluid was used as a template to clone and sequence the genome of SARS-CoV-2. Shortly after, the World Health Organization announced a worldwide pandemic. However, later reports revealed that the COVID-19 pathogen has a genetic footprint that had never been observed in natural coronavirus, suggesting a "gain of function virus" that might have been engineered in a laboratory. Clear conclusions about the outbreak's origins are still lacking. Theories regarding most probable wildlife animal reservoir for the SARS-CoV-2 have been scientifically shaky. Exactly how the virus jumped from wild animals to humans remains a mystery. Onging debate coalesces around 2 competing theories: laboratory accidental versus intentional escape scenario of a genetically engineered virus versus zoonotic emergence. Within a year after the declaration of the COVID-19 pandemic, Pfizer-BioNTech and Moderna published their 2-month duration follow-up mRNA vaccine trial results. Later reports revealed a rapidly declining vaccineinduced immunity, questionable effectiveness of these vaccines against the new variants despite boosting, inability to stop viral transmission, and, most importantly, the onset of serious adverse events. In the 2 context of all these uncertainties, controversies, and the remaining many unanswered questions, this first review presents a different perspective on the origin of the COVID-19 pandemic. A later analysis will address the subsequent events that followed in terms of diagnosis, protective measures, vaccines roll-out and vaccine mandate.

A Different Perspective on the COVID-19 Pandemic: Validity of COVID-19 Testing and Protective Nonpharmaceutical Interventions (Part 2).

The causative agent for COVID-19 was identified by next-generation sequencing from samples received from several pneumonia patients. The detection of the coronavirus was done through real-time polymerase chain reaction analyses performed from plasma sample. Ribonucleic acid extracted from a patient's bronchoalveolar lavage fluid was used as a template to clone and sequence a genome from lineage B of the betacoronavirus. A polymerase chain reaction test was considered positive using cycle threshold above 34 for detected samples. Tests with thresholds so high will detect genetic leftover fragments, which pose no particular risk. No correlation between viral load and disease severity has ever been connected to a virus. On the basis of the little science centered on misleading mortality calculations and most importantly on mathematical disease modeling, generalized lockdowns were imposed on a global scale, with a considerable number of fatalities and resulting in global economic breakdown. Most studies found little to no evidence for the effectiveness of face masks in the general population, neither as personal protective equipment nor as a source control. In this review, we attempted to expose a different perspective of some aspects of the pandemic related to disease diagnosis and protective nonpharmacologic interventions. Our viewpoint raises serious questions on the validity of such restrictive measures in saving lives and warn against the application of such damaging strategies in a potential future outbreak.

Bronchial aspirate obtained during bronchoscopy yields increased fungal load compared to bronchoalveolar lavage fluid in patients at risk of invasive aspergillosis and Pneumocystis pneumonia.

Bronchoalveolar lavage fluid (BALF) is a standard respiratory sample for diagnosing invasive fungal diseases like Pneumocystis pneumonia (PCP) and invasive pulmonary aspergillosis (IPA). However, procedural variations exist across medical centers and wards. This study aimed to compare the diagnostic potential of BALF and bronchial aspirate (BA) obtained during bronchoscopy in 173 patients suspected of fungal infections. A prospective observational study was conducted from April 2020 to November 2021. BALF and BA were collected during bronchoscopy and subjected to direct examination, fungal culture, Aspergillus fumigatus qPCR (AfqPCR), and Pneumocystis jirovecii qPCR (PjqPCR). Galactomannan detection was performed on BALF. Patients were classified based on established European Organization for Research and Treatment of Cancer (EORTC) criteria. Out of 173 patients, 75 tested positive for at least one test in BA or BALF. For Aspergillus, proportion of positive AfqPCR (14.5% vs. 9.2%; P<0.0001) and fungal loads (Cq of 31.3 vs. 32.8; P=0.0018) were significantly higher in BA compared to BALF. For Pneumocystis, fungal loads by PjqPCR was also higher in BA compared to BALF (Cq of 34.2 vs. 35.7; P=0.003). BA only detected A. fumigatus and P. jirovecii in 12 (42.9%) and 8 (19.5%) patients, respectively. BA obtained during a BAL procedure can be a suitable sample type for increased detection of P. jirovecii and A. fumigatus by qPCR. The use of BA in diagnostic algorithms requires further investigation in prospective studies.

The application of nanopore targeted sequencing for pathogen diagnosis in bronchoalveolar lavage fluid of patients with pneumonia: a prospective multicenter study

Objective To evaluate the value of nanopore targeted sequencing in diagnosing pneumonia pathogens. Methods This large-scale multicentre prospective study performed in 8 hospitals across China from April to October 2022. Hospitalised patients with a diagnosis of pneumonia at admission were included. Complete clinical data were collected, and bronchoalveolar lavage fluid were obtained from each patient. These samples underwent simultaneous testing using conventional microbial testing, metagenomic next-generation sequencing, and nanopore targeted sequencing. Results A total of 218 patients were included. Among the 168 cases of pulmonary infection, 246 strains of pathogens were confirmed. Nanopore targeted sequencing outperformed conventional microbial testing, identifying more pathogens with a sensitivity increase of 47.9% (77.2% vs. 29.3%). Metagenomic next-generation sequencing had a sensitivity of 82.9%. Total of 70.1% patients had consistent results in both metagenomic next-generation sequencing and nanopore targeted sequencing. Nanopore targeted sequencing exhibited significantly higher sensitivity in detecting Pneumocystis jiroveci, cytomegalovirus, Mycobacterium tuberculosis, Nontuberculous mycobacteria, Streptococcus pneumoniae, and Mycoplasma pneumoniae compared to conventional microbial testing. However, metagenomic next-generation sequencing demonstrated higher sensitivity than nanopore targeted sequencing for Aspergillus (88.5% vs. 53.8%). Regarding the detection of co-infections, nanopore targeted sequencing displayed significantly higher sensitivity than conventional microbial testing (76.7% vs. 28.7%) and was on par with metagenomic next-generation sequencing (76.7% vs. 82.9%). Conclusion Nanopore targeted sequencing performs equally well as metagenomic next-generation sequencing in bronchoalveolar lavage fluid for pathogen diagnosis in pneumonia, both methods showing higher sensitivity than conventional microbial testing. Nanopore targeted sequencing can be considered a reliable method for diagnosing pathogens in pneumonia.

Flow Cytometric Analysis of Macrophages and Cytokines Profile in the Bronchoalveolar Lavage Fluid in Patients with Lung Cancer.

Macrophages play an important role in the suppression and activation of immune anti-cancer response, but little is known about dominant macrophage phenotype in the lung cancer environment, evaluated by bronchoalveolar lavage fluid (BALF). The aim of this study was to characterize macrophages in BALF from a lung affected by cancer (cBALF) and a healthy lung (hBALF) of the same patient regarding their individual macrophage polarization and selected cytokines profile. A total of 36 patients with confirmed lung cancer were investigated. Macrophages markers: CD206 CD163 CD80 CD86 CD40 CD45, Arginase-1, and CD68 were evaluated by flow cytometry. Cytokines (IL-1 RA, IL-6, IL-10, TNF-α, IL-1β, IL-12, IL-23, and TGF-β) profile was analyzed. There was higher median proportion of macrophages in Cbalf than in Hbalf. The population of macrophages presented immunophenotype: Ccd68+bright CD206+bright CD163+bright CD80+ CD86+ CD40+bright CD45+ cArginase+. We observed some trends in the expression of the analyzed antigens in clBALF and hlBLAF. The highest concentrations of IL-1RA and IL-6 were in Cbalf and Hbalf supernatant. There were the correlations between pro- and anti-inflammatory cytokines. The findings showed that macrophages include a diverse and plastic group with the presence of different antigens and cytokines, and determining the target phenotype is a complex and variable process.

Expression analysis of inflammatory factors in artificial quartz stone plate processing silicosis patients

Objective: To investigate the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1beta (IL-1β) in the plasma and bronchoalveolar lavage fluid of silicosis patients with artificial quartz stone plate processing. Methods: In January 2022, 10 patients with artificial quartz stone plate processing silicosis and 20 patients with common silicosis who were hospitalized and diagnosed in a hospital at Zhejiang Province from June 2019 to December 2021 were retrospectively selected as the research objects, and 30 healthy people were selected as the control group during the same period. Plasma of all subjects and bronchoalveolar lavage fluid of all patients were collected. The levels of TNF-α, IL-6 and IL-1β in plasma and bronchoalveolar lavage fluid were detected by enzyme-linked immunosorbent assay and were analyzed. Results: The levels of TNF-α, IL-6 and IL-1β in the plasma of patients with silicosis were higher than those of the control group (P<0.05), and the levels of TNF-α and IL-1β in the plasma of silicosis patients with artificial quartz stone plate processing were higher than those of common silicosis patients (P<0.05). The levels of TNF-α and IL-1β in plasma of artificial quartz stone plate processing silicosis patients were higher than those of common silicosis patients at the same silicon stage (P<0.05). The levels of IL-1β in bronchoalveolar lavage fluid of silicosis patients with artificial quartz stone plate processing was higher than that of patients with common silicosis (P<0.05) . Conclusion: The levels of TNF-α, IL-6 and IL-1β in silicosis patients with artificial quartz stone plate processing are higher than those in patients with common silicosis, which may be related to dust components they are exposed to.

IL18R1-Related Molecules as Biomarkers for Asthma Severity and Prognostic Markers for Idiopathic Pulmonary Fibrosis.

To determine the role of inflammation-related proteins in predicting asthma severity and outcome, 92 inflammation-related proteins were measured in the asthmatic serum using Olink analysis. Different bioinformatics algorithms were developed to cross analyze with the single-cell or transcriptome data sets from the Gene Expression Omnibus database to explore the role of IL18R1 and related genes in asthma and idiopathic pulmonary fibrosis (IPF). Olink identified 52 differentially expressed proteins in asthma. They were strongly linked to the cytokine-cytokine receptor interaction, TNF, and NF-κB signaling pathway. Seven proteins were found in both single-cell RNA and Olink analyses. Among them, IL18R1 was predominantly expressed in mast cells, and the results suggested enhanced communication between mast cells and CD 8+ T cells. IL18R1 was upregulated in serum and induced sputum and bronchoalveolar lavage fluid of patients with uncontrolled or severe asthma. IL18R1 was positively correlated with TNFSF1 and OSM and S100A12. The diagnostic efficacy of these serum IL18R1-related molecules for asthma ranged from 0.839 to 0.921. Moreover, high levels of IL18R1, TNFSF1, OSM, and S100A12 were significantly associated with shorter survival times and worse lung function. IL18R1-related molecules may serve as biomarkers for monitoring uncontrolled or severe asthma and as prognostic markers for IPF.

Analysis of cytokines in bronchoalveolar lavage fluid in patients with checkpoint inhibitor pneumonitis and pulmonary infection: A case-control study.

The discovery of immune checkpoint inhibitors (ICIs) is a breakthrough in the field of cancer therapy. However, ICIs may cause immune-related adverse reactions, including checkpoint inhibitor-related pneumonitis (CIP). The aim of this study was to investigate cytokines in bronchoalveolar lavage fluid (BALF) of patients with CIP compared with patients with pulmonary infection and patients with cancer. We retrospectively analyzed 34 cytokine levels and T cell subsets in BALF supernatant samples from ICI-treated patients with CIP (n = 13), pulmonary infection (n = 10), and progressive cancer (n = 12). Cytokine levels and T cell subsets were compared among the three groups of patients. We observed significantly higher levels of IFN-γ-induced protein 10(IP-10) (p = 0.002), and percentage of CD3 + CD8 + T cells (p = 0.020) in BALF of patients with CIP compared with the other two groups. However, we found significantly lower levels of interleukin-21 (p = 0.008) in BALF of patients with progressive disease compared with the other two groups. Cytokine profile and character of cell subsets in BALF was helpful for the differential diagnosis of CIP. IP-10 may play an important role in pathophysiology for CIP and also be a potential therapeutic target.

Comparative study of eosinophil count in peripheral blood, sputum and BAL fluid in patients of bronchial asthma

Bronchial Asthma is defined by the history of respiratory symptoms such as wheeze, shortness of breath, chest tightness and cough that vary over time and in intensity, together with variable expiratory airflow limitation. Airflow limitation may later become persistent over the course of disease. To compare the findings of eosinophil counts in peripheral blood, induced sputum and bronchoalveolar lavage fluid in asthmatic patients and healthy individuals. This Case Control Cross sectional study was conducted among patients attending respiratory OPD at Sir Sunder Lal Hospital, BHU, Varanasi, with diagnosis of bronchial asthma (100) and healthy controls. Significant association was found between eosinophilic bronchial asthma and absolute eosinophil count (W = 1168.000, p = 0.020), total serum IgE (W = 1338.000, p = &amp;#60;0.001), BAL eosinophil count (χ2 = 94.589, p = &amp;#60;0.001), sputum eosinophil count (χ2 = 14.057, p = &amp;#60;0.001).

Diagnostic accuracy of cryptococcal antigen test in pulmonary cryptococcosis: a protocol for a systematic review and meta-analysis

BackgroundThe cryptococcal antigen (CrAg) test was proposed as a rapid diagnostic tool to identify cryptococcal meningitis in patients suffering from AIDS. Several studies have demonstrated its diagnostic performance in cryptococcal...

Severe Pneumocystis Pneumonia Associated with Ulcerative Colitis

A 56-year-old woman with a 9-month history of ulcerative colitis (UC) who had been treated with prednisolone (PSL) 40 mg/day and azathioprinefor 2 months was referred to our hospital because of fever and dyspnea. Computed tomography (CT) of the chest revealed diffuse ground-glass opacities in both lungs and consolidation in both lower lobes. PCR of the patient's bronchoalveolar lavage fluid was positive for pneumocystis jirovecii, and she was diagnosed with pneumocystis pneumonia (PCP). The patient developed refractory respiratory failure that required invasive mechanical ventilation. After sulfamethoxazole-trimethoprim administration and methyl prednisolone pulse therapy, her clinical condition improved dramatically. Few reports are available on severe PCP associated with UC. It is important to keep in mind the possibility of complication by PCP in UC patients.

A Preliminary Study on Microbiota Characteristics of Bronchoalveolar Lavage Fluid in Patients with Pulmonary Nodules Based on Metagenomic Next-Generation Sequencing

The characteristics and roles of microbes in the occurrence and development of pulmonary nodules are still unclear. We retrospectively analyzed the microbial mNGS results of BALF from 229 patients with pulmonary nodules before surgery, and performed a comparative analysis of lung flora between lung cancer and benign nodules according to postoperative pathology. The analysis also focused on investigating the characteristics of lung microbiota in lung adenocarcinomas with varying histopathology. There were differences in lung microbiota between lung cancer and benign lung nodules. Bacterial diversity was lower in lung cancer than in benign lung nodules. Four species (Porphyromonas somerae, Corynebacterium accolens, Burkholderia cenocepacia and Streptococcus mitis) were enriched in lung cancer compared with the benign lung nodules. The areas under the ROC curves of these four species were all greater than 0.6, and the AUC of Streptococcus mitis was 0.702, which had the highest diagnostic value for differentiating lung cancer from benign lung diseases. The significantly enriched microbiota varied with the different pathological subtypes of lung adenocarcinoma. Streptococcus mitis, Burkholderia oklahomensis and Burkholderia latens displayed a trend of increasing from the benign lung disease group to the AIS group, MIA group and IAC group, whereas Lactobacillus plantarum showed a downward trend. Changes in the abundance of lung microbiota are closely related to the development of infiltrating adenocarcinoma. Our findings provide new insights into the relationship between the changes in lung microbiota and the development of lung cancer.

The Relationship Between Oral Microbiota and Chronic Obstructive Pulmonary Disease

Oral microbiota have a complex impact on the host's health and disease states. It has been found that the composition of lung flora bears a striking resemblance to the composition of oral flora. Moreover, oral pathogenic bacteria have been detected in the sputum and bronchoalveolar lavage fluid of patients with chronic obstructive pulmonary disease (COPD), suggesting that oral microbiota play an important role in the pathogenesis and development of COPD. Findings from lots of studies have shown that oral microbiota may participate in the pathogenesis and development of COPD through non-specific immune response, specific immune response, and the activities of protein hydrolase. Herein, we mainly summarized the available evidence on the relationship between oral microbiota and COPD. By examining the relationship between the two, we elaborated on the application of oral microbiota in the diagnosis and prevention of COPD, discussed possible directions for future research, and provided references for developing new therapeutic approaches.

Bacteriological Analysis of Bronchoalveolar Lavage Fluid in Patients with Respiratory Tract Infections

Background and Aim: Bacterial respiratory infections are most commonly causes of illness for all age group patientsin ICU. Most of the patients suffer from urosepsis, postoperative disease and lower respiratory infection whenadmitted in ICU’s. Broncho alveolar lavage (BAL) is an ideal sample that allows the recovery of pathogens cellularand noncellular components from the epithelial surface of lower respiratory tract. This study was performed todetect pathogenic organisms by microscopy of BAL fluid and isolate and identify various bacteria and fungi fromBAL fluid in culture and analyze their antibiogram.Material and Methods: The cross-sectional prospective study was conducted in the Department of Microbiology,tertiary care teaching hospital, India. The study included 200 BAL samples taken from all consecutive patientsreferred with suspicion of pneumonia. Bronchial wash was done with the help of fibreoptic bronchoscope underlocal anaesthesia. All BAL samples were cultured on three bacteriological media agar plates using a sterile 4mmnichrome loop (0.01ml), and incubated at 37 C for 72 hours for quantitative bacterial culture using standardlaboratory techniques. Bacterial isolates were identified by performing standard microbiological procedures suchas study of colony morphology, Gram staining and standard biochemical tests.Results: Out of the total 200 samples, 120 (60%) were from males, and 80 (40%) were female patients. Thepredominant GNB was Klebsiella pneumoniae 45 (61%), followed by Pseudomonas aeruginosa 22(30%), Esch.coli 6(8%) and the fungal isolate was Aspergillus niger 5(1%). Klebsiella &amp; Pseudomonas were highly sensitive toamikacin, piperacillin-tazobactam, imipenem, gentamycin, followed by tobramycin.Conclusion: Results of the present study demonstrate the high incidence of gram-negative isolates. The study alsosuggests that regular antimicrobial sensitivity monitoring should be done as most isolates are highly resistant tocephalosporin and other commonly used antimicrobials

Metagenomic next-generation sequencing as an unconventional approach to warn of tumor cells in a patients with non-mucinous pneumonic-type lung adenocarcinoma: Case report.

Pneumonic-type lung cancer (PTLC) is a special type of lung cancer with cough and expectoration as the main clinical symptoms and inflammatory signals as the main imaging manifestations. PTLC can be easily misdiagnosed as pneumonia, and the diagnosis and treatment are always delayed. Metagenomic next-generation sequencing (mNGS), as an emerging and effective method to identify occult pathogens, has been gradually adopted by clinicians. A 58-year-old woman with recurrent cough and expectoration was admitted to hospital on January 12th, 2022. She reported that she was diagnosed with pneumonia half a month ago, after treatment with expectorant and antibiotics for 5 days, the symptoms were relieved. However, the symptoms worsened again 10 days after stopping the drugs. On the current presentation, she denied exposure to patients with infection of COVID-19, smoking history, night sweats, weight loss, rash, joint pain, fever, and shortness of breath. The patient was diagnosed with non-mucinous pneumonic-type lung adenocarcinoma according to the clinical symptoms, changes of CT scans after treatment and cytopathology examinations. The patient was initially diagnosed with pulmonary infection according to computerized tomography (CT) scan. Expectorant and antibiotics used. However, the symptoms worsened again 10 days after stopping the drugs. On her return visit, the CT scan did not showed obvious consolidation absorption and was similar to the previous imaging findings. mNGS was performed to detect the occult pathogens. None pathogen was detected, however, 39 copy number variations were found in Human Chromosomal Instability Analysis of mNGS indicating the presence of tumor cells. The cytopathology findings confirmed the presence of lung adenocarcinoma (non-mucinous adenocarcinoma). She was treated with targeted antitumor drugs, and the CT scan after 20 days of targeted antitumor therapy showed obvious absorption of the lesions. mNGS may have potential value to screen tumor cells in bronchoalveolar lavage fluid of patients with PTLC, especially in the patients whose samples in bronchioli cannot be collected using existing sampling tools.

Targeting epithelial cell-derived TWIST1 alleviates allergic asthma

It is well known that the T Helper (Th)2 bias plays a critical role in allergic asthma. Whereas the Th2 bias is maintained in the local tissues is uncertain. IL-33 is vital for the development of the Th2 polarization. TWIST-1 has an effect on regulating cellular functions. The aberrant activation of RAS sustains certain cellular activities. The aim of this study is to study the role of the interaction between activation of TWIST1 and RAS in inducing and maintaining Th2 polarization in allergic asthma. The epithelial cells of the airways (AEC) were isolated from the broncho-alveolar lavage fluids in patients with asthma. The mediators involved in the over-expression of IL-33 were determined by RNA sequencing. A mouse model was established to test the role of TWIST1 and RAS in developing allergic asthma. We observed a strong expression of TWIST1 in patients with allergic asthma that showed a positive correlation with asthmatic responses. TWIST1 favored the expression of the IL-33 in the AEC. Twist1-deficient AEC-carrying mice did not induce Th2 polarization in the airways. The expression TWIST1 in AECs was positively associated with RAS activation in AECs in patients with allergic asthma. The interaction between RAS and TWIST1 in AECs sustained airway allergic inflammation. Inhibition of TWIST1 or RAS prevented asthma-like inflammation in the mouse airways. In summary, the interaction between TWIST1 and RAS induces and maintains IL-33 expression in AECs to facilitate allergic inflammation in the respiratory tract. Inhibition of TWIST1 or RAS can prevent experimental allergic asthma.