Bronchoalveolar lavage fluid assessment facilitates precision medicine for lung cancer

Lung cancer is the most common and fatal malignant disease worldwide and has the highest mortality rate among tumor-related causes of death.Early diagnosis and precision medicine can significantly improve the survival rate and prognosis of lung cancer patients.At present,the clinical diagnosis of lung cancer is challenging due to a lack of effective non-invasive detection methods and biomarkers,and treatment is primarily hindered by drug resistance and high tumor heterogeneity.Liquid biopsy is a method for detecting circulating biomarkers in the blood and other body fluids containing genetic information from primary tumor tissues.Bronchoalveolar lavage fluid(BALF)is a potential liquid biopsy medium that is rich in a variety of bioactive substances and cell components.BALF contains information on the key characteristics of tumors,including the tumor subtype,gene mutation type,and tumor environment,thus BALF may be used as a diagnostic supplement to lung biopsy.In this review,the current research on BALF in the diagnosis,treatment,and prognosis of lung cancer is summarized.The advantages and disadvantages of different components of BALF,including cells,cell-free DNA,extracellular vesicles,and micro RNA are introduced.In particular,the great potential of extracellular vesicles in precision diagnosis and detection of drug-resistant for lung cancer is highlighted.In addition,the performance of liquid biopsies with different body fluid sources in lung cancer detection are compared to facilitate more selective studies involving BALF,thereby promoting the application of BALF for precision medicine in lung cancer patients in the future.

Bronchoalveolar lavage fluid metagenomic next-generation sequencing assay for identifying pathogens in lung cancer patients

Background:For patients with lung cancer,timely identification of new lung lesions as infectious or non-infectious,and accurate identification of pathogens is very important in improving OS of patients.As a new auxiliary examination,metagenomic next-generation sequencing(mNGS)is believed to be more accurate in diagnosing infectious diseases in patients without underlying diseases,compared with conventional microbial tests(CMTs).We designed this study tofind out whether mNGS has better performance in distinguishing infectious and non-infectious diseases in lung cancer patients using bronchoalveolar lavagefluid(BALF).Materials and Methods:This study was a real-world retrospective review based on electronic medical records of lung cancer patients with bronchoalveolar lavage(BAL)and BALF commercial mNGS testing as part of clinical care from 1 April 2019 through 30 April 2022 at The First Affiliated Hospital of Sun Yat-sen University.164 patients were included in this study.Patients were categorized into the pulmonary non-infectious disease(PNID)group(n=64)and the pulmonary infectious disease(PID)group(n=100)groups based onfinal diagnoses.Results:BALF mNGS increased the sensitivity rate by 60%compared to CMTs(81%vs.21%,p<0.05),whereas there was no significant difference in specificity(75%vs.98.4%,p>0.1).Among the patients with PID,bacteria were the most common cause of infection.Fungal infections occurred in 32%of patients,and Pneumocystis Yersini was most common.Patients with Tyrosine kinase inhibitors(TKIs)therapy possess longer overall survival(OS)than other anti-cancer agents,the difference between TKIs and immuno-checkpoint inhibitors(ICIs)was insignificant(median OS TKIs vs.ICIs vs.Anti-angiogenic vs.Chemo vs.Radiotherapy=76 vs.84 vs.61 vs.58 vs.60).Conclusions:our study indicates that BALF mNGS can add value by improving overall sensitivity in lung cancer patients with potential pulmonary infection,and was outstanding in identifying Pneumocystis infection.It could be able to help physicians adjust the follow-up treatment to avoid the abuse of antibiotics.

Pneumocystis jirovecii diagnosed by next-generation sequencing of bronchoscopic alveolar lavage fluid: A case report and review of literature

BACKGROUND The advent of molecular targeted agents and immune checkpoint inhibitors has greatly improved the treatment of advanced renal cell carcinoma(RCC), thus significantly improving patient survival. The incidence of rare drug-related adverse events has gained increased attention.CASE SUMMARY We report a patient with advanced RCC treated with multiple lines of molecular targeted agents and immune checkpoint inhibitors, who developed a pulmonary infection after treatment with everolimus in combination with lenvatinib. Determining the pathogenic organism was difficult, but it was eventually identified as Pneumocystis jirovecii by next-generation sequencing(NGS) of bronchoscopic alveolar lavage fluid(BALF) and successfully treated with trimethoprim-sulfamethoxazole.CONCLUSION Rare pulmonary infections caused by molecular targeted agents are not uncommon in clinical practice, but their diagnosis is difficult. Evaluating BALF with NGS is a good method for rapid diagnosis of such infections.

Value of eosinophil count in bronchoalveolar lavage fluid for diagnosis of allergic bronchopulmonary aspergillosis: A case report

BACKGROUND An immediate hypersensitive immune response to Aspergillus fumigatus antigens is one of the main characteristic features of allergic bronchopulmonary aspergillosis(ABPA).As ABPA is an allergic respiratory disease,immunoglobulin E and peripheral-blood eosinophilia have been used as diagnostic indicators.However,eosinophilia in bronchoalveolar lavage fluid(BALF)has not been considered in the diagnostic criteria for ABPA.CASE SUMMARY We present a case of ABPA in which the eosinophil count in peripheral blood was not increased,whereas the eosinophil percentage in BALF reached 60%.After antifungal and hormone therapy,imaging revealed very good resolution of lung infiltration.CONCLUSION The value of the eosinophil count in BALF for the diagnosis of ABPA is worthy of the clinician's attention,especially when the patient’s clinical features lack specificity and the diagnostic parameters are negative.

Characteristics of alveolar macrophages in bronchioalveolar lavage fluids from active tuberculosis patients identified by single-cell RNA sequencing

Tuberculosis(TB),is an infectious disease caused by Mycobacterium tuberculosis(M.tuberculosis),and presents with high morbidity and mortality.Alveolar macrophages play an important role in TB pathogenesis although there is heterogeneity and functional plasticity.This study aimed to show the characteristics of alveolar macrophages from bronchioalveolar lavage fluid(BALF)in active TB patients.Single-cell RNA sequencing(scRNA-seq)was performed on BALF cells from three patients with active TB and additional scRNA-seq data from three healthy adults were established as controls.Transcriptional profiles were analyzed and compared by differential gene expression and functional enrichment analysis.We applied pseudo-temporal trajectory analysis to investigate correlations and heterogeneity within alveolar macrophage subclusters.Alveolar macrophages from active TB patients at the single-cell resolution are described.We found that TB patients have higher cellular percentages in five macrophage subclusters.Alveolar macrophage subclusters with increased percentages were involved in inflammatory signaling pathways as well as the basic macrophage functions.The TB-increased alveolar macrophage subclusters might be derived from M1-like polarization state,before switching to an M2-like polarization state with the development of M.tuberculosis infection.Cell-cell communications of alveolar macrophages also increased and enhanced in active TB patients.Overall,our study demonstrated the characteristics of alveolar macrophages from BALF in active TB patients by using scRNA-seq.

Stability of Xuesaitong capsules in gastrointestinal lavage fluid

Objective:To obtain a formulation with high bioavailability through evaluation of the stability of three types of Xuesaitong capsules in the stomachs and intestines of rats.We compared the stability of the Panax notoginsenoside R1 as well as the ginsenosides Rg1,Rb1,Re,and Rd in different formulations.Methods:Artificial stomach fluid(ASF)and artificial intestinal fluid(AIF)were prepared.Stability of three types of Xuesaitong capsules was examined for 4 h in stomachs and 24 h in intestines.Samples were analyzed at different times by high-performance liquid chromatography.Percent content of NGR1,GRg1,GRb1,GRe,and GRd at different times was calculated.Results:Hard capsules incubated in ASF disintegrated within 2e3 min,whereas soft capsules disintegrated within 7e8 min.Components in hard capsules were dissolved rapidly in water,with content of each compound reaching 90%in 5 min,and degradation of each compound reaching 30e50%after incubation for 240 min.Dissolution and degradation of each component in soft capsules with a water-soluble base tended to balance at 30 e90 min.Contents in soft capsules with a lipid-soluble base showed slow dissolution after ASF incubation for 120 min.Five saponins in identical types of capsules incubated in ASF had similar stability curves.Contents of hard capsules and soft capsules with a watersoluble base degraded rapidly within 30 min and reached a plateau when Xuesaitong capsules were incubated in AIF.

Effects of early bronchoalveolar lavage fluid collected from dogs with smoke inhalation injury on the lungs of rats

Objective: Whether early massive bronchoalveolar lavage can remove the harmful substances from the lungs injured with smoke inhalation remains uncertain. This study was designed to observe the effects of early massive bronchoalveolar lavage fluid (BALF) on the healthy lungs in rats. Methods: Mongrel dogs were inflicted with severe smoke inhalation injury. The injured lungs were lavaged with large amount of normal saline in the first hour after injury and the BALF was collected. The BALF was injected into the healthy lungs of 30 rats (group C) in the dosage of 5 ml/kg. The functions and pathological changes of the lungs were observed 24 h after perfusion with the BALF. The data were compared with those of 23 rats (group B) whose lungs were perfused with the BALF collected from normal dogs and those of 21 rats (group A) whose lungs were perfused with normal saline. Results: The mortality rate 24 h after lung perfusion was higher in group C than in groups A and B. The survivors of group C exhibited fluctuation of respiratory rate (RR), remarkable decrease of PaO 2, significantly higher content of lung water, decrease of total static pulmonary compliance and pulmonary expansion index, and increasse of inflammatory cytokines in the tissues of lungs. Only slight mechanic obstructive effect on the airway was observed in rats of group A and B. The pathological changes of the lungs of the rats in group C were similar to those of the dogs with actual smoke inhalation injury. Conclusion: Our findings indicate that the BALF collected from dogs with acute severe smoke inhalation injury in the early stage after injury injured the normal lungs of rats with the bioactive substances in the BALF. These findings show us that it is a valuable therapeutic procedure to apply massive bronchoalveolar fluid lavage in the early stage after inhalation injury.

Expression of endostatin in serum and bronchoalveolar lavage fluid in patients with lung cancer

Objective To evaluate the expression of endostatin in bronchoalveolar lavage fluid (BALF) and serum in patients with lung cancer,and to analysis the relationship between endostatin expression and clinical prognosis as well as pathophysiological characteristics. Methods The samples of serum and BALF were obtained from 57

XpertMTB/RIF Assay for the Diagnosis of Pulmonary Tuberculosis Based on Bronchoalveolar Lavage Fluid:A Systematic Review and Meta-analysis

Currently,tuberculosis(TB)is the second most lethal disease in the world caused by a single infectious pathogen.Rapid diagnosis of TB is of great importance for its treatment and management.Xpert MTB/RIF is a novel rapid diagnostic assay for the diagnosis of pulmonary TB(PTB).Use of the Xpert assay based on bronchoalveolar lavage fluid(BALF)samples is indicated when TB is suspected and sputum smears or cultures are negative.The aim of this meta-analysis was to systematically evaluate the diagnostic performance of the Xpert assay based on BALF samples for the diagnosis of PTB.A systematic review of previously published articles was performed,and rel-evant data were extracted.Meta-DiSc 1.4 and Stata 12.0 were used to analyze the data.When Mycobacterium tuberculosis cultures were used as the criterion standard,the combined sensitivity of BALF-based Xpert was 0.89(95%CI,0.87–0.91),the specificity was 0.87(95%CI,0.85-0.88),the positive likelihood ratio was 8.28(95%CI,5.39–12.71),the negative likelihood ratio was 0.14(95%CI,0.10–0.19)and the diagnostic ratio was 84.08(95%CI,42.00–168.31).When composite reference standard was used as the criterion standard,the above observations were 0.69(95%CI,0.67–0.72),0.98(95%CI,0.97–0.98),41.40(95%CI,14.56–117.71),0.28(95%CI,0.21–0.37)and 190.47(95%CI,50.56–717.54),respectively.The area under the summary receiver operating characteristic curve was close to 1 for both.Overall,the Xpert MTB/RIF assay based on BALF samples showed high sensitivity and specificity for the diag-nosis of PTB and seems to be a reliable rapid detection method.

Analysis of the T cell receptor Vδ region gene repertoire in bronchoalveolar lavage fluid (BALF) and peripheral blood of asthmatics

Objective To explore the role of γδT cells in the airway of asthmatics and to identify the forces which induce and maintain the inflammatory processMethods Peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) were obtained from7 asthmatic subjects and 7 nonsmoker control subjects The percentage of γδT cells in the PB and BALF was measured by immunofluorescent staining and flow cytometry The frequency of usage and the clonality of Vδ subfamilies (Vδ 1-Vδ 3) were assessed by RTPCR and gene scanning Results A higher proportion of γδT cell was detected in the BALF of asthmatic subjects (7.8%±4.7%) than that from control subjects (3.3%±3.0%, P=0.04) No selective usage for a particular Vδ subfamily was found, but the relative expression level of Vδ 1 was significantly higher in the asthmatic airway (44%±13%) than in the control (19%±5%, P=0.0002) In asthmatic subjects, the monoclonal or oligoclonal expansion of γδT lymphocytes was predominant in the BALF, especially Vδ 1+ T lymphocytes Conclusions Antigenic specific γδT cells might play an important role in the inducement and maintenance of airway inflammation Persistent antigenic stimulation may be the key factor that maintains chronic airway inflammation in asthma

Alteration of surfactant proteins A and D in bronchoalveolar lavage fluid of pneumocystis carinii pneumonia

Objective To understand the interaction between surfactant proteins and pneumocystis carinii pneumonia (PCP),and the impact of corticosteriods on surfactant proteins.Methods We established rat models of PCP and bacterial pneumonia induced by subcutaneous injection of 25mg cortisone acetate.At 8- 12 wk,the bronchoalveolar lavage fluid(BALF)of rats was collected.Total nucleated cells of BALF were counted and differentiated,and the concentrations of surfactant protein A(SP-A)and surfactant protein D(SP-D)were measured by immunoblotting assay.The rats were divided into three immunosuppressive groups and a normal control group.Group I,normal control(n = 6),consisted of healthy SD rats;group Ⅱ,negative control(n = 6),consisted of rats with cortisone acetate injection for over 8 wk without lung infection;group Ⅲ,bacterial pneumonia(n = 11),rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated;and group Ⅳ,PCP(n = 14),rats with injected cortisone acetate for 8 - 12 wk and developed PCP without other pathogens isolated.Results Our results indicated that the total cell count in BALF in the negative control group was lower than that in the normal control group(P ＜ 0.001).During PCP infection,the total cell count and the percentage of polymorphonuclearcytes(PMNs)in BALF were significantly increased(P ＜ 0.01),but were lower than those in the bacterial pneumonia group.The concentration of SP-A of BALF in PCP(45.1 ± 22.1 μg/ml)was significantly increased in comparison with that in the negative control(16.2 ± 9.9 μg/ml,P ＜ 0.05)and bacterial pneumonia groups(6.2 ± 5.6 μg/ml,P ＜ 0.001).We also found that the relative content of SP-D was significantly higher in PCP(24249 ±4780 grey values)than that in the negative control (13 384 ± 2887 grey values,P ＜ 0.001)and that in bacterial pneumonia(11 989 ± 2750 grey values,P＜0.001).SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group(P ＜ 0.01,P ＜ 0.001).SP-A and SP-D were higher in the negative control group than those in the normal control group,but there was no significant difference between the 2 groups.Conclusion These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation,but the alteration is not related to the corticosteriod usage.

Distribution of Major Pathogens from Sputum and Bronchoalveolar Lavage Fluid in Patients with Noncystic Fibrosis Bronchiectasis： A Systematic Review

Objective： Noncystic fibrosis （non-CF） bronchiectasis remains as a common health problem in Asia. Pathogens＇ distribution in airways of patients with non-CF bronchiectasis is important for doctors to make right decision. Data Sources： We performed this systematic review on the English language literatures from 1966 to July 2014, using various search terms included ＂pathogens＂ or ＂bacteria＂ or ＂microbiology＂ and ＂bronchiectasis＂ or ＂non-cystic fibrosis bronchiectasis＂ or ＂non-CF bronchiectasis＂ or ＂NCFB.＂ Study Selection： We included studies of patients with the confirmed non-CF bronchiectasis for which culture methods were required to sputum or bronchoalveolar lavage fluid （BALF）. Weighted mean isolation rates for Haemophilus influenzae, Pseudomonas aeruginosa, Streptococcus pneumoniae, Stapylococcus aureus, Moxarella catarrhails were compared according to different methodology. Results： The total mean bacterial culture positive rates were 63%. For studies using sputum samples, the mean positive culture rates were 74%. For studies using BALF alone or BALF and sputum, it was 48%. The distributions of main bacterial strains were 29% for 1-1. influenzae, 28% for P. aeruginosa, 11% for S. pneumoniae, 12% for S. aureus, and 8% for M. catarrhails with methodology of sputum. Meanwhile, the bacterial distributions were 37% for H. influenzae, 8% for P aeruginosa, 14% for S. pneumoniae, 5% for S. attretts, and 10% for M. catarrhails with methodology of BALF alone or BALF and sputum. Analysis of the effect of different methodology on the isolation rates revealed some statistically significant differences. Conclusions： H. influenzae accounted for the highest percentage in different methodology. Our results suggested that the total positive culture rates and the proportion of P. aeruginosa from sputum and BALF specimens had significant differences, which can be used in further appropriate recommendations for the treatment of non-CF bronchiectasis.

Indicators for prediction of Mycobacterium tuberculosis positivity detected with bronchoalveolar lavage fluid

Background:The diagnosis of active pulmonary tuberculosis(TB)remains a challenge in clinic,especially for sputum negative pulmonary TB.Bronchoalveolar lavage fluid(BALF)has higher sensitivity than sputum for detection of Mycobacterium tuberculosis(Mtb).However,bronchoscopy is invasive and costly,and not suitable for all patients.In order to make TB patients get more benefit from BALF for diagnosis,we explore which indicator might be used to optimize the choice of bronchoscopy.Methods:A total of 1539 sputum-smear-negative pulmonary TB suspects who underwent bronchoscopy were recruited for evaluation.The sensitivity,specificity and accuracy of Mtb detection in sputum and BALF were compared.Odds ratios and 95%confidence intervals were used to assess variables that associated with positive acid-fast bacilli(AFB)smear,Mtb culture and nucleic acid amplification test(NAAT)of BALF in sputum-negative and non-sputum-producing pulmonary TB suspects.Results:BALF has significantly higher sensitivity(63.4%)than sputum(43.5%)for Mtb detection by culture and NAAT.19.7%(122/620)sputum-negative and 40.0%(163/408)non-sputum-producing suspects had positive bacteriological results in BALF.Among sputum-negative and non-sputum-producing pulmonary TB suspects,the positivity of Mtb detection in BALF is associated with a younger age,the presence of pulmonary cavities and a positive result of interferon-gamma release assay(IGRA).Sputum-negative patients under 35 years old with positive IGRA and pulmonary cavity had 84.8%positivity of Mtb in BALF.Conclusions:Our study indicated that combination of age,the presence of pulmonary cavity,and the result of IGRA is useful to predict the positivity of Mtb detection in BALF among sputum-negative and non-sputum producing pulmonary TB suspects.Those who are under 35 years old,positive for the presence of pulmonary cavity and IGRA,should undergo bronchoscopy to collect BAFL for Mtb tests,as they have the highest possibility to get bacteriologically confirmation of TB.

Microscopic examination of intracellular organisms in bronchoalveolar lavage fluid for the diagnosis of ventilator-associated pneumonia：a prospective multi-center study

Background The presence of intracellular organisms （ICOs） in polymorphonuclear leukocytes obtained from bronchoalveolar lavage fluid （BALF） is a possible method for rapid diagnosis of ventilator-associated pneumonia （VAP）.However,the validity of this diagnostic method remains controversial and the diagnostic thresholds reported by investigators were different.Our objective was to evaluate the accuracy of quantification of ICOs in BALF for the diagnosis of VAP,and to detect the best cutoff percentage of PMNs containing ICOs （PIC） in the microscopic examination of BALF for the diagnosis of VAP.Methods This was a prospective multi-center study conducted in 4 ICUs in Wuhan,China,which involved 181 patients suspected of first episode of VAP.BALF was obtained from all enrolled patients.The BALF samples underwent quantitative culture,cytological and bacteriological analysis to detect the culture results,PIC values and the morphological features of microorganisms.Definite diagnosis of VAP was based on pre-set criteria.The receiver-operating characteristic curve was used to detect the best cutoff point for PIG to diagnose VAP,and the diagnostic accuracy was calculated.Moreover,quantitative culture and Gram＇s stain of BALF were adopted to diagnose VAP,and their diagnostic accuracy was evaluated as well.Results There were 102 patients definitely diagnosed with VAP （VAP group）,and 60 patients definitely diagnosed without VAP （no VAP group）.We found that ICOs were present in 96.08％ （98 out of 102） of VAP patients and 20.00％ （12 out of 60） of no VAP patients.The PICs were significantly higher （（9.53±6.65）％ vs.（0.52±1.33）％,P＜0.01） in VAP group.In our study,the best cutoff point for PIC to diagnose VAP was 1.5％,which had a sensitivity of 94.12％,a specificity of 88.33％,a positive predictive value （PPV） of 93.20％ and a negative predictive value （NPV） of 89.83％.The area under the receiveroperating characteristic curve was 0.956 （95％ confidence interval,0.925-0.986; P＜0.01）.When the positive quantitative culture results of BALF were used to diagnose VAP,the sensitivity,specificity,PPV and NPV were 65.69％,95.00％,95.71％ and 61.96％,respectively.Whereas they were 70.59％,76.67％,83.72％ and 60.53％,respectively,when the positive Gram＇s stain results of BALF were used to diagnose VAP.The concordance between the results of Gram＇s stain and quantitative cultures was poor,only 32.10％ （52 out of 162） was totally right,and 17.28％ （28 out of 162） was partially right.Conclusions PIC＞1.5％ has good diagnostic performance in the microscopic examination of BALF for the diagnosis of VAP.However,Gram＇s stain is not reliable for the early application of antibiotic therapy,due to the poor bacteriological predictive value.

Cellular changes in bronchoalveolar lavage fluid in hyperoxiainduced lung injury

It is well known that high concentration oxy-gen exposure is a model of acute lung injury(ALI).However,controversy exists over the mechanism.This study was designed to clarify the cellular characteristics in bronchoalveolar lavage fluid(BALF)and body weight loss of rats exposed to oxygen(.90%).Young male Wistar rats,aged 6 weeks,were divided into three groups:(1)room air group(exposed to room air,n 522);(2)hyperoxia,48 h group(exposed to over 90%oxygen for less than 48 h,n 518);(3)hyperoxia 66–72 h group(exposed to over 90%oxygen for 66–72 h group,n 57).Compared to the room air group,the total cell counts in the hyperoxia 66–72 h group decreased,whereas the neu-trophils increased significantly.The body weights of the rats exposed to room air continued to increase.However,the body weights of oxygen-exposed rats increased slightly on the first day and weight loss was seen from the second day.All rats were noted to have bilateral pleural effusion in the hyperoxia 66–72 h group.The data suggests that(1)an increase in neutrophil count is an evident feature of hyperoxia-induced lung injury;(2)high concentration oxygen exposure can give rise to anorexia and malnutri-tion,which may play a role in hyperoxia-induced lung injury.Blocking neutrophil influx into lung tissue in the early phase and improving malnutrition are two effective methods to reduce hyperoxic lung injury.

The clinical significance of increased neutrophils in bronchoalveolar lavage fluid in sarcoidosis

Objective To investigate the clinical implications of increased neutrophils in bronchoalveolar lavage fluid(BALF)in sarcoidosis.Methods A retrospective analysis was performed for 72 cases of histologically diagnosed,treatment-nive sarcoidosis admitted to Peking University Third Hospital from January 2000 to October2014.The patients included 18 men and 54 women。

Pulmonary abscess caused by Streptococcus pseudopneumoniae in a child:A case report and review of literature

BACKGROUND Lung abscess found on chest X-ray and computed tomography examinations is rare in infants and young children.Several pathogens can cause lung abscesses,with the most common pathogens being anaerobes,Streptococci and Staphylococcus aureus.Streptococcus pseudopneumoniae(S.pseudopneumoniae)is a member of the Streptococcaceae family,and is mainly isolated from respiratory tract specimens.There are currently no cases of lung abscess caused by S.pseudopneumoniae in the literature.CASE SUMMARY A 2-year-old boy was admitted to hospital due to persistent cough and fever.Lung computed tomography examination suggested the formation of a lung abscess.His diagnosis was not confirmed by testing for serum respiratory pathogens(6 items),respiratory pathogen nucleic acid(27 items),and laboratory culture.Finally,metagenomic next-generation sequencing of bronchoalveolar lavage fluid revealed the presence of S.pseudopneumoniae,confirming its role in causing the lung abscess.After receiving antibiotic treatment,reexamination with lung computed tomography showed that the abscess was resorbed and the patient’s outcome was good.CONCLUSION This is the first report of a lung abscess in a child caused by S.pseudopneumoniae infection.Metagenomic next-generation sequencing of bronchoalveolar lavage fluid is helpful in achieving rapid and accurate pathogen identification.

Protection of Allergic Asthma in Mice by Black Rice Bran Bioprocessed with Shiitake Mushroom Mycelia

We evaluated the potential of orally fed new food formulations to inhibit biomarkers reported to be involved in the causes of allergic asthma in mice. Asthma, a serious non-communicable disease, affects both adults and children and can be undertreated. New functional foods could provide therapeutic approaches. Here, the anti-asthma mechanism of a new functional food and three isolated fractions produced by bioprocessing black rice bran with shiitake mushroom mycelia was evaluated in mast cells, B cells, and orally fed mice and compared with non-bioprocessed black rice bran. In vitro, the treatments inhibited RBL-2H3 cell degranulation and immunoglobulin E (IgE) production. The in vitro anti-asthma effects were confirmed in orally fed mice following asthma induction by alumina and chicken egg ovalbumin (OVA). The suppression of asthma resulted from the inhibition of inflammation- and immune-related substances, including OVA-specific IgE, thymic stromal lymphopoietin, eotaxin, leukotriene C4, prostaglandin D2, and vascular cell adhesion molecule-1 in bronchoalveolar lavage fluid and serum. The treatment also reversed the thickening of the lung airway wall. The inflammation and asthma inhibition seems to be regulated by the balance of the T-helper cells’ Th1/Th2 immune response and the inhibition of multiple biomarkers associated with the cause of asthma. Future human clinical studies with adults and children should determine the potential therapeutic value of the anti-asthma effects of the new functional foods.

Role of RECK methylation in gastric cancer and its clinical significance

AIM:To investigate the relation between RECK methylation and clinicopathological characteristics of gastric cancer patients and evaluate the role of RECK methylation in peritoneal metastasis of gastric cancer.METHODS:Methylation of RECK gene in 40 paired samples of gastric cancer and its corresponding adjacent normal mucosa,lymph nodes and peritoneal irrigation fluid was detected by methylation-specific polymerase chain reaction.RESULTS:Aberrant methylation of RECK gene was detected in 27.5%(11/40)of the adjacent normal mucosa samples,in 47.5%(19/40)of gastric cancer samples,in 57.1%(12/21)of the lymph node samples,and in 35%(14/40)of peritoneal irrigation fluid samples,respectively,with a significant difference between the adjacent normal mucosa and lymph node samples(P=0.023).Presence of RECK methylation in the primary tumor samples was significantly correlated with tumor invasion(P=0.023).The accuracy of RECK methylation in peritoneal lavage fluid samples for the diagnosis of peritoneal metastasis of gastric cancer was 72.5%(26/40),with a sensitivity of 66.7%(6/9) and a specificity of 74.2%(23/31).CONCLUSION:Aberrant methylation of RECK gene may provide useful information for the early diagnosis and treatment of peritoneal metastasis of gastric cancer.

Clinical Features and Prognosis in 21 Patients with Extrinsic Allergic Alveolitis

Objective To assess the spectrum of causes, clinical features, differences between disease phases, and prognosis of extrinsic allergic alveolitis （EAA）. Methods Patients with EAA diagnosed at Peking Union Medical College Hospital from August 1983 to May 2007 were analyzed retrospectively. Their medical records were examined to gather clinical, laboratorial, radiological, and histopathological data. Patients were divided to three phases （acute, subacute, and chronic） according to clinical presentations. Follow-up data regarding treatment response, subsequent radio- logical and pulmonary function studies, and clinical outcomes were collected. Results A total of 21 cases were enrolled. Among them, 11 were subacute, 10 were chronic. The most common exposure was pet birds （6 cases, 28.6%）. The primary abnormality of pulmonary function was restriction and/or reduction in diffusing capacity （12 cases, 63.2%）. The most common findings on high-resolution computed tomography （HRCT） were ground-glass opacities （13 cases, 68.4%） and centri- lobular nodules （8 cases, 42.1%）. Airway obstruction in pulmonary function test, emphysema, hmg cysts, and fibrosis on HRCT were more frequently seen in chronic than in subacute patients, though the differences were not statistically significant. Bronchoalveolar lavage fluid （BALF） showed lymphocytosis. The total＇cell count and the percentage of neutrophils were significantly higher in subacute than in chronic patients （P〈0.05）. Nonnecrotizing granulomas were seen in 8 （47.1%） cases. Improvement or normalization in symptoms, radiography, and pulmonary function test after treatment were seen in all 18 patients with avail- able follow-up data. Five patients recurred. Conclusions The characteristic abnormalities of pulmonary function, findings on HRCT, and pa- thology are essential for all phases of EAA, and the atypical manifestations such as obstruction and fibrosis can also be present frequently, particularly in chronic cases. Differential cell counts of BALF are related to the phase of the disease. The treatment response and prognosis of EAA are good.