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.

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.

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.

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.

Comparison of LAMP,GeneXpert,Mycobacterial Culture,Smear Microscopy,TSPOT.TB,TBAg/PHA Ratio for Diagnosis of Pulmonary Tuberculosis

Objective:To investigate the application value of loop-mediated isothermal amplification(LAMP),GeneXpert,mycobacterial culture,smear microscopy,TSPOT.TB(TSPOT),ratio of TB-specific antigen to phytohemagglutinin(TBAg/PHA ratio)in the detection of mycobacterium tuberculosis in the bronchoalveolar lavage fluid.Methods:A retrospective analysis was performed on the patients who underwent bronchoscopy from December 2018 to November 2019 in Tongji Hospital.The patients with positive tuberculosis culture or positive GeneXpert in bronchoalveolar lavage fluid were selected as the case group,and those without tuberculosis served as the control group.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of LAMP,GeneXpert,culture,smear microscopy,TSPOT,and TBAg/PHA ratio.Results:For the patients with positive cultures as case,the sensitivity of LAMP,GeneXpert,smear microscopy,TSPOT and TBAg/PHA ratio was 73.49%,89.16%,25.30%,80.00%,33.85%,respectively,the specificity was 99.00%,100.00%,99.00%,86.00%,100.00%,respectively,the area under the ROC curve(AUC)was 0.849,0.938,0.633,0.830,0.669,respectively.For the patients with positive GeneXpert as case,the sensitivity of LAMP,mycobacterial culture,smear microscopy,TSPOT and TBAg/PHA ratio was 73.20%,74.23%,22.68%,68.92%,29.73%,respectively,the specificity was 99.00%,100.00%,99.00%,86.00%,100.00%,respectively,the AUC was 0.853,0.878,0.623,0.775,0.649,respectively.Conclusion:The sensitivity of GeneXpert was best.The sensitivity and diagnostic value of LAMP were slightly lower than those of GeneXpert,and were similar to tuberculosis culture.The sensitivity of smear microscopy was low.The specificity of TSPOT was low.When TBAg/PHA ratio>0.2 was used as a diagnostic index,the specificity was improved,but the sensitivity was low.

Biochemical Assessment of Acute Lung Injury Induced by Hexavalent Chromium Compound

Acute pulmonary injury triggered by sodium chromate (Na 2CrO 4) and it′s recovery duration were evaluated by analysing the biochemical responses in bronchoalveolar lavage fluids(BALF) in Sprague Dawley rats. The animals of five groups that received single intratracheal instillations of Na 2CrO 4 suspension at doses of 0.008, 0.04, 0.20, 0.98 and 4.88 mg·kg -1 body wt respectively were killed at d 2 after treatment in the dose response experiment. The animals of seven groups that received the instillations of 0.98 mg·kg -1 Na 2CrO 4 were sacrificed at d 2, 4, 7, 10, 14, 21 and 28 separately after exposure in the time course experiment. The biochemical indicators examined in the BALF included N acetyl β D glucosaminidase (NAG), gamma glutamyl transferase (GGT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), angiotensin converting enzyme (ACE), α 1 antitripsin(A1AT), albumin (Alb) and lysozyme(LYS). Based on the present findings, it was found that no effect dose and threshold dose of Na 2CrO 4 were 0.008 and 0.04 mg·kg -1 respectively and that the majority of biochemical parameters in the BALF showed the dose dependent and significant increases at dosed of ≥0.20 mg·kg -1 . And at the severe intoxication dose ≥0.98 mg·kg -1 all parameter values could significantly be increased, which lasted about 1 week and the recovery duration needed at least 4 weeks. The present study indicates that ≥0.98 mg·kg -1 Na 2CrO 4 can result in acute lung injury including pulmonary cyto biomembrane damage, diffuse alveolitis and edema, and that NAG, LYS, ALP, Alb and ACE are probably the valuable and early sensitive indicators for evaluating the injury after exposure to lower dose Na 2CrO 4 (<0.98 mg·kg -1 ).

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 DETECTION AND CLINICAL SIGNIFICANCE OF hEGF AND β＿2－MG IN BALF OF PATIENTS WITH LUNG CANCER

Human epidermal growth factor (hEGF) and β2-Microglohulin(β2-MG) in bronchoalveolar lavage fluid (BALF) and serum were detected in 74 patients with lung cancer and 76 patients with benign lung disease. The results showed that the positive rates or both hEGF (58.1%) and β2-MG (62. 2%) in BALF of the affected side of lung cancer were higher than those of either the healthy side of lung cancer or benign lung disease (P<0.05), and also higher than that in serum.The level of BALF-hEGF was highest in squamous cell carcinoma and adenocarcinoma, and it was higher in stages Ⅲ, Ⅳ than in stages Ⅰ, Ⅱ. The combinative detection or hEGF and β2-MG in BALF and serum will he valuable to early diagnosis of lung cancer.

Treatment of acute lung injury in mice using Bai-Ri-Ke syrup

Objective:This study aimed to examine the therapeutic effects of Bai-Ri-Ke syrup(BRK)on mice with acute lung injury(ALI).Methods:Fifty male C57BL/6 mice were equally divided into the control group,model group,dexamethasone(DXM)group,Bai-Ri-Ke syrup(BRK)low-group,and BRK-high group,with six mice per group.An intratracheal injection of 5 mg/kg POLY(I:C)was used to construct an ALI mouse model.After a successful model construction,the mice in the DXM group were given[0.2 mg/10 g·d]dexamethasone sodium phosphate injection(1 mL:2 mg)on the following day via intraperitoneal injection.The mice in the BRK-low group were given 0.015 mL APS everyday by gavage,and the mice in the BRK-high group were given 0.030 mL APS everyday by gavage for three days.The wet to dry weight(W/D)ratio of the lungs was observed every day.Bronchoalveolar lavage fluids(BALFs)were collected from the left lungs to measure the BALF protein level and neutrophil count after 72 h of treatment.The IL-6,IL-1β,and TNF-αlevels in BALF were also measured.HE staining was done to observe the histopathological changes in the lungs.Results:The ALI mice in the BRK-high group had significantly increased W/D(P<0.01).ELISA results showed that the DXM group and BRK-high group had significantly decreased BALF protein content(P<0.01),neutrophil count(P<0.01),and IL-6,IL-1β,and TNF-αlevels(P<0.01).Hematoxylin and eosin(H&E)results showed that the DXM group and BRK-high group had alleviated alveolar tissue injury,edema,bleeding,and inflammation.Conclusions:The BRK can decrease the W/D,BALF protein content,neutrophil count,and TNF-α,IL-1β,and IL-6 levels and alleviate the histopathological changes in the lungs of ALI mice.

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。

Host protection against Omicron BA.2.2 sublineages by prior vaccination in spring 2022 COVID-19 outbreak in Shanghai

The Omicron family of SARS-CoV-2 variants are currently driving the COVID-19 pandemic.Here we analyzed the clinical laboratory test results of 9911 Omicron BA.2.2 sublineages-infected symptomatic patients without earlier infection histories during a SARS-CoV-2 outbreak in Shanghai in spring 2022.Compared to an earlier patient cohort infected by SARS-CoV-2 prototype strains in 2020,BA.2.2 infection led to distinct fluctuations of pathophysiological markers in the peripheral blood.In particular,severe/critical cases of COVID19 post BA.2.2 infection were associated with less pro-inflammatory macrophage activation and stronger interferon alpha response in the bronchoalveolar microenvironment.Importantly,the abnormal biomarkers were significantly subdued in individuals who had been immunized by 2 or 3 doses of SARS-CoV-2 prototypeinactivated vaccines,supporting the estimation of an overall 96.02% of protection rate against severe/critical disease in the 4854 cases in our BA.2.2 patient cohort with traceable vaccination records.Furthermore,even though age was a critical risk factor of the severity of COVID-19 post BA.2.2 infection,vaccination-elicited protection against severe/critical COVID-19 reached 90.15% in patients aged≥60 years old.Together,our study delineates the pathophysiological features of Omicron BA.2.2 sublineages and demonstrates significant protection conferred by prior prototype-based inactivated vaccines.

Sleep-related hypoxemia aggravates systematic inflammation in emphysematous rats

Background Sleep disturbance is common in patients with emphysema. This study aimed to develop a novel model of sleep-related hypoxemia （SRH） in emphysema （SRHIE） with rats, and to explore the inflammatory status of SRHIE in lung, liver, pancreas, carotid artery and whole blood.Methods Seventy-five male Wistar rats were assigned to 5 groups with 15 per group according to the exposure conditions. The protocols varied with the degree of hypoxia exposure and severity of pre-existing emphysema caused by cigarette smoke exposure： （1） SRH control （SRHCtrl） group, sham smoke exposure （smoke exposure, exposed to smoke of 15 cigarettes twice everyday, 16 weeks） and SRH exposure （12.5% O2, 3 hours, SRH exposure, divide total hypoxia time （1.5 hours or 3 hours） into 4 periods evenly （22.5 minutes or 45 minutes） and distribute these hypoxia periods evenly into physiological sleep time of rats identified by electroencephalogram, week 9 to week 16）; （2） Emphysema control （ECtrl） group, smoke exposure and sham SRH exposure （21% O2, 3 hours）; （3） Short SRH in emphysema （SRHShort） group, smoke exposure and short SRH exposure （12.5% O2, 1.5 hours）; （4） Mild SRH in emphysema （SRHMild） group,smoke exposure and mild SRH exposure （15% O2, 3 hours）; （5） Standard SRH in emphysema （SRHStand） group, smoke exposure and SRH exposure （12.5% O2, 3 hours）. ECtrl, SRHShort, SRHMild and SRHStand groups were groups with emphysematous rats. Two days before the end of exposure, 5 rats in each group were randomly selected for arterial blood gas analysis. In the rest 10 rats in each group, we obtained blood samples and bronchoalveolar lavage fluid （BALF）for routine tests. We also obtained tissue blocks of lung, liver, pancreas, and right carotid artery for pathologic scoring and measurements of liver oxidative stress （measuring hepatic oxidative stress enzymes, superoxide dismutase （SOD） activity, catalase （CAT） activity and malondialdehyde （MDA） concentration）.Results Emphysematous groups had higher mean linear intercept （MLI） and mean alveolar number （MAN） values than SRHCtrl group. MLI values in SRHStand group were the highest （ail P 〈0.05）. O2Sat in SRHStand rats when SRH exposure was （83.45±1.76）%. Histological scores of lung, liver, pancreas and right carotid artery were higher in emphysematous groups than SRHCtrl group, and SRHStand group were the highest （all P 〈0.05） （SOD and CAT values were lower and MDA values were higher in groups with emphysema than without and in SRHStand group than in ECtrl group （all P 〈0.05））. MDA values were the highest in SRHStand group （all P 〈0.05）. Total cellular score in BALF and White blood cell （WBC） in whole blood were the highest in SRHStand group （all P 〈0.05）. Lymphocyte ratios were the highest in SRHStand group both in BALF and blood （all P 〈0.05）. Red blood cell （RBC） and hemoglobin in emphysematous groups were higher than that in SRHCtrl group, and SRHStand group were higher than ECtrl group （all P 〈0.05）.Conclusions With a proper novo model of SRHIE with Wistar rats, we have demonstrated SRH may aggravate the degree of emphysematous changes, polycythemia,oxidative stress and systematic inflammation. SRH and emphysema may have a synergistic action in causing systematic damages, and lymphocyte may be playing a central role in this process. Longer duration and more severe extent of SRHIE exposure also seem to result in more serious systematic damages. The mechanisms of all these concerned processes remain to be studied.

Efficacy of Yiqigubiao pill on chronic obstructive pulmonary disease in rats with the disease induced by lipopolysaccharide and cigarette-smoke fumigation

OBJECTIVE:To investigate the efficacy of Yiqigubiao pill(YQGB)on chronic obstructive pulmonary disease(COPD)in rats with the COPD induced by lipopolysaccharide(LPS)and cigarettesmoke fumigation.METHODS:In this study,six groups of rats were set up,including control group,model group,positive control group(aminophylline)and YQGB(high,medium and low doses)groups.Tracheal injection of lipopolysaccharide(LPS)and cigarette-smoke fumigation induced COPD in rats.The general condition,incubation period and coughing times,lung function,level of inflammatory factors,leukocyte condition and pathological changes of bronchus and lung tissue were observed in rats of each group.RESULTS:In the COPD rats,the latent period of coughing was shortened and the cough frequency was increased significantly;the pulmonary function was significantly decreased,which was manifested by the increased lung tissue resistance and respiratory system resistance,and the decreasing percentage of forced expiratory volume and forced expiratory volume in the 0.3 s(FEV0.3/FVC);the contents of tumor necrosis factor-alpha(TNF-α)and interleukin-4 in serum were obviously increased,and the NEUT%in bronchoalveolar lavage fluid was significantly increase.YQGB could obviously prolong the latent period of cough,and reduce the cough frequency and the content of TNF-αin serum.YQGB can also significantly reduce respiratory resistance and increase FEV0.3/FVC value.The results of histopathology showed that YQGB significantly reduced the pathological changes of tracheal mucosa and lung caused by COPD.YQGB obviously increased level of AQP1,which was down-regulated in the COPD rats.CONCLUSION:YQGB could significantly improve the pulmonary function,reduce inflammation and alleviate lung and bronchial diseases in the COPD rats.

Relationship between bone marrow-derived CD_(34)^+ cells expressing interleukin-5 receptor messenger RNA and asthmatic airway inflammation

BACKGROUND: Asthma is clinically related with the degree of eosinophilic inflammation. How asthmatic airway inflammation is affected is still poorly understood. So the effects of bone marrow-derived hematopoietic cells expressing CD(34) (CD(34)(+)) and interleukin-5 (IL-5) receptor messenger RNA (IL-5R mRNA+) on asthmatic airway inflammation were investigated. METHODS: Balb/c mice were sensitized and challenged by ovalbumin (OVA) to establish an asthmatic model while control mice were sensitized and exposed to sterile saline. The mice were killed at different time points after being challenged by OVA and sterile saline. Then, bronchoalveolar lavage fluid (BALF), peripheral blood (PB) and bone marrow (BM) were prepared. Eosinophils in PB (PBEOS) and BALF (BALFEOS), nuclear cells in BALF, PB and BM were counted. By flow cytometry, the percentage of CD(34)(+) cells to nucleated cells in PB, BM and the relative number of CD(34)(+) cells in PB (PBCD(34)(+)) and BM (BMCD(34)(+)) were calculated. Immunocytochemistry and in situ hybridization were used to investigate the hematopoietic cells with co-localized expression of CD(34) and IL-5R mRNA in BM (BMCD34+IL-5R mRNA+). The percentage of BMCD34+IL-5R mRNA+ to BMCD(34)(+) was calculated. RESULTS: Twelve hours after challenge by OVA, BALFEOS and PBEOS in the experimental group were significantly higher than those in the control group (P

Effect of radix paeoniae rubra on expression of p38 MAPK/iNOS/HO-1 in rats with lipopolysaccharide·induced acute lung injury

Objective: To investigate the effect of radix paeoniae rubra (RPR) on expression of p38 mitogen activated protein kinase ( MAPK )/iNOS/HO-1 in rats with lipopolysaccharide-induced acute lung injury and explore the molecular mechanism.Methods: Forty healthy male Wistar rats, weighing 200-250 g, aged 6-8 weeks (mean =7 weeks), provided by the Experimental Center, Medical College, Wuhan University, Wuhan, China, were employed in this study.Under anesthesia with 7% chloraldurat (5 ml/kg body weight) through intraperitoneal injection, the trachea of the rat was exposed and an arterial puncture needle pricked into the trachea via cricothyroid membrane. Then they were randomly divided into five groups: 8 rats receiving 1 ml normal saline through the puncture needle (Group A),8 receiving 1 ml lipopolysaccharide (LPS, 2.5 mg/kg,Group B), 8 receiving LPS and RPR (30 mg/kg, pumped through the femoral vein for 2 hours, Group C ), 8 receiving RPR 2 hours before dripping LPS ( Group D),and 8 receiving hemin (75 μmol/L through intraperitoneal injection) 18 hours before dripping LPS (Group E). After 6 hours of LPS dripping, blood samples were obtained through the carotid artery to perform blood gas analysis,then all the rats were exsanguinated to death and specimens of lung tissues were obtained. The pathomorphological changes of the lung tissues were observed. The expression of p38 MAPK/iNOS/HO-1, the neutrophil ratio, protein content in alveolar irrigating solution and malonaldehyde (MDA) content in the lung tissues were also detected.Results: Compared with Group A, the expression of p38 MAPK, iNOS and HO-1 markedly increased in Groups B, C, D, and E (P ＜0.01). But in Groups C, D, and E,the expression of p38 MAPK and iNOS were significantly lower than that of Group B, while expression of HO-1 was obviously higher than that of Group B ( P ＜ 0.05 ). The protein content, the ratio of neutrophils in bronchoalveolar lavage fluid ( BALF), the content of MDA and the activities of serum NO in Group B were significantly higher than those of Group A ( P ＜ 0.01 ). There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Group B (P ＜ 0.01). Compared with Group B, these indexes of lung injury were significantly lower while the levels of arterial bicarbonate and partial pressure of oxygen increased significantly in Groups C, D,and E (P ＜ 0.05 or P ＜ 0.01 ). Under light microscope, the pathological changes induced by LPS were significantly attenuated by RPR and hemin.Conclusions: The high expression of MAPK plays an important role in lipopolysaccharide-induced acute lung injury. Protective effect of RPR on lipopolysaccharideinduced acute lung injury may be related to the inhibition of the abnormal high expression of p38 MAPK/iNOS/HO-1.

Comparison of Composition and Diversity of Bacterial Microbiome in Human Upper and Lower Respiratory Tract

INTRODUCTIONMicrobiome residing in the airways and parenchymal tissues, as a biological barrier of respiratory tract, plays an important role in maintaining the normal functions of individual＇s respiratory system and preventing the invasion and colonization of exogenous pathogens.H1 Changes of microbial community may result in the occurrence and progression of infectious pneumonia and acute exacerbation of chronic diseases, through disordering commensal microorganisms and increasing invasive ability of pathogens.F21 Traditional methods for detecting bacteria normally rely on culture technique, which have a number of critical problems, including time-consuming and relatively low positive rate of detection. In the last 1 0 years, 16S rRNA gene sequencing technique has been thought to be more comprehensive and reliable than those of culture-dependent methods for monitoring bacterial microbiome in skin, organs, and tracts quantitatively and qualitatively.E31 In this case, choosing credible samples are critical to evaluate the community composition of microbiome in the host tracts, including respiratory tract. In the present study, we aimed to compare the composition and diversity of bacterial community structure in different samples collected from the individuals＇ airways using 16S rRNA gene sequencing technique.