Bronchoalveolar Lavage Fluid Of Children Research Articles

The correlation between the level of cellular classification in bronchoalveolar lavage fluid of children with severe Mycoplasma pneumoniae and clinical characteristics.

The BALF of children with SMPP is predominantly neutrophilic. A lower percentage of lymphocytes is related to a higher incidence of pleural effusion, extrapulmonary manifestations and progression to RMPP, as well as a longer length of hospitalisation. • Mycoplasma pneumonia in children is relatively common in clinical practice. Bronchoalveolar lavage (BAL) is a routine clinical procedure. However, there are relatively few studies focusing on the cytomorphological analysis of cells in BAL fluid.

Oxidative stress in the alveolar lavage fluid of children with Mycoplasma pneumoniae pneumonia.

To investigate the correlation between oxidative stress in the bronchoalveolar lavage fluid (BALF) of children with Mycoplasma pneumoniae pneumonia (MPP) and the clinical characteristics of severe MPP (SMPP) and refractory MPP (RMPP). Clinical and BALF-related data were collected from 83 patients with MPP, of which 29 had SMPP and 54 had general MPP (GMPP); 37 patients were in the RMPP group and 46 in the non-RMPP group. The levels of malondialdehyde (MDA) and advanced oxidation protein products (AOPP) as well as the activity levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in BALF were detected. Logistic regression analyses were performed on MDA, AOPP, SOD, GSH-PX, gender, heat peak, neutrophil percentage, C-reactive protein, lactate dehydrogenase, d-dimer, lung consolidation, sputum embolus, and pleural effusion. The levels of MDA and AOPP in the BALF of the MPP group were significantly higher than those in the control group (p < .05), whereas SOD and GSH-PX levels were lower than those in the control group (p < .05). The BALF AOPP levels in the RMPP group were higher than those in the non-RMPP group, and the SOD and GSH-PX levels in the BALF were lower than those in the non-RMPP group; the difference was statistically significant (p < .05). The levels of MDA and AOPP in the BALF of children in the SMPP group were higher than those in the GMPP group, and the levels of SOD and GSH-PX were lower than those in the GMPP group, with statistically significant differences (p < .05). The C-index of the logistic regression model was 0.960 (95% confidence interval 0.958-0.963), which indicates that the model has good predictive ability. Advanced oxidation protein products may be a marker for predicting the conditions of SMPP and RMPP, and the prediction model can assess the risk of progression in children to RMPP, which is conducive to clinical diagnosis and treatment.

Changes in Cellular Morphology in Bronchoalveolar Lavage Fluid of Children with Mycoplasma Pneumoniae Pneumonia

Objective To study changes of cell morphology in BALF in children with Mycoplasma pneumoniae pneumonia (MPP). Methods From December 2021 to May 2022, a group of 32 children diagnosed with Acute MPP and admitted for treatment in the Pediatrics Department and PICU of the First Affiliated Hospital of Xinxiang Medical University were selected for our study. These patients underwent bronchoalveolar lavage as part of their clinical assessment. For comparison, we included a control group comprising 10 children who were not infected but had bronchial foreign bodies. We investigated the cellular composition in the bronchoalveolar lavage fluid (BALF) using Wright-Giemsa staining and microscopic evaluation, aiming to understand the relationship between shifts in cell proportions and extra-pulmonary symptoms associated with MPP. Results In this study, a total of 42 cases were enrolled, with 32 cases in the study group and 10 cases in the control group. There were no statistically significant differences in gender, age, height, weight, and BMI between the two groups (p &gt; 0.05). The study group exhibited significantly higher levels of neutrophil percentage (GRA%), CRP, D-dimer, and LDH in blood routine tests compared to the control group (p &lt; 0.05). Furthermore, the proportions of neutrophils (%) and macrophages (%) in BALF were significantly higher in the study group compared to the control group (p&lt; 0.05), while the proportion of lymphocytes (%) in BALF showed no statistically significant difference between the two groups (p&gt; 0.05). Conclusion In the acute phase of MPP in children, BALF is predominantly composed of neutrophils. A lower proportion of lymphocytes in BALF is associated with a higher incidence of extra-pulmonary manifestations and longer hospitalization duration.

IL-27 Levels in Bronchoalveolar Lavage Fluid in Children with Post-infectious Bronchiolitis Obliterans.

Pulmonary neutrophils may play a crucial role in the development of bronchiolitis obliterans (BO) following measles virus infection. IL-27 could potentially have a negative regulatory effect on the release of reactive oxygen species and cytotoxic granules in neutrophils. To investigate the levels of IL-27 in the bronchoalveolar lavage fluid (BALF) of children with post-infectious bronchiolitis obliterans (PIBO) and analyze the relationship between IL-27 levels and neutrophil proportions. A total of 24 children with PIBO were recruited for the experimental group, while 23 children with bronchial foreign bodies were included in the control group. Bronchoscopic alveolar lavage was performed in both groups. The levels of IL-27 in BALF were measured using enzyme-linked immunosorbent assay (ELISA). The proportions of neutrophils in BALF were determined by smear staining. The relationship between the levels of IL-27 in BALF and the neutrophil proportions was analyzed by the Pearson test. The levels of IL-27 in BALF were significantly lower in children with PIBO compared to children with bronchial foreign bodies (p<0.05). Additionally, the proportions of neutrophils in BALF were significantly higher in children with PIBO compared to children with bronchial foreign bodies (p<0.05). The levels of IL-27 were negatively correlated with the neutrophil proportions in BALF in children with PIBO (p<0.05), but not in children with bronchial foreign bodies (p>0.05). The present study suggests that a decrease in IL-27 may be associated with an increase in neutrophils in BALF and may contribute to the pathogenesis of PIBO.

Analysis of cytokine levels, cytological findings, and MP-DNA level in bronchoalveolar lavage fluid of children with Mycoplasma pneumoniae pneumonia.

The present study was conducted to determine the inflammatory response in the lungs of children with Mycoplasma pneumoniae pneumonia (MPP). This study retrospectively analyzed cytokine levels, cytological findings, and M. pneumoniae (MP)-DNA level in the bronchoalveolar lavage fluid (BALF) of 96 children with MPP. The study utilized Spearman's correlation method to evaluate the contribution of BALF and blood parameters in MPP children. (1) A total of 96 MPP children were classified into the Low MP-DNA MPP group (BALF MP-DNA ≤ 105 copies/mL) and the High MP-DNA MPP group (BALF MP-DNA > 105 copies/mL); the Non-fever MPP group (no fever during the entire course of pneumonia) and the Fever MPP group; the Defervescence MPP group (fever had subsided at the time of bronchoscopy) and the Fervescence MPP group; and the Mild MPP group and the Severe MPP group. (2) The High MP-DNA MPP, Fever MPP, Fervescence MPP, and Severe MPP groups had higher levels of interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) in their BALF (all p < .05). (3) The proportions of neutrophils and macrophages in the BALF of the High MP-DNA MPP and Fever MPP groups increased and decreased, respectively (all p < .05). (4) In the BALF of MPP children, MP-DNA, IL-6, IL-10, TNF-α, and interferon gamma (IFN-γ) levels positively correlated with neutrophil proportion while negatively correlated with macrophage proportion (all p < .05). (5) The MP-DNA, IL-6, IL-10, TNF-α, and IFN-γ levels in the BALF of MPP children were positively correlated with the levels of C-reactive protein, procalcitonin, lactic dehydrogenase, fibrinogen, and d-dimer, while they were negatively correlated with the albumin level (all p < .05). In children with MPP, the pulmonary inflammatory immune response was stronger in the High MP-DNA MPP, Fever MPP, Fervescence MPP, and Severe MPP groups. The relationship between pulmonary cytokine levels, MP-DNA load, and serum inflammatory parameters were found to be weak.

Correlation between Mycoplasma pneumoniae drug resistance and clinical characteristics in bronchoalveolar lavage fluid of children with refractory Mycoplasma pneumoniae pneumonia

BackgroundTo investigate the resistance-gene mutation of Mycoplasma pneumoniae (MP) in the bronchoalveolar lavage fluid of children with Mycoplasma pneumoniae pneumonia (MPP) and the clinical characteristics of refractory Mycoplasma pneumoniae pneumonia (RMPP) correlation.MethodsForty-eight children with MPP were selected and placed in RMPP and non-RMPP groups based on their clinical status – whether they had worsening clinical symptoms, persistent fever and a worsening lung image. They were also separated into drug-resistance gene mutation and non-mutated groups using nucleic acid detection. The participants’ data were collected on high-sensitivity C-reactive protein and MP-DNA loads, fever time, hospitalisation time, macrolide antibiotic application time and fever regression time after application. The differences in imaging manifestations were determined by using multivariate logistic regression to analyse the clinical characteristics of RMPP. Additionally, the correlation between drug-resistance gene mutations and the clinical characteristics of RMPP was summarised.ResultsAmong the 48 MPP children, 31 (64.6%) had A2063G and/or A2064G gene mutation, 31 (64.6%) had RMPP and 23 (74.2%) had drug-resistance gene mutation. The children in the drug-resistance gene mutation group had higher high-sensitivity C-reactive protein and MP-DNA loads, longer fever time, hospitalisation time, macrolide antibiotic application time, fever regression time after application and extrapulmonary complications. There were more symptoms and more severe changes under bronchoscopy. The difference was statistically significant (P < 0.05). Logistic multivariate regression analysis showed that the mutation of drug-resistance genes had no significant correlation with RMPP.ConclusionThe mutation rate of drug-resistance genes in children with MPP is high, the inflammatory index and MP-DNA load are high, the course of the disease is long, and the changes under bronchoscopy are severe. The occurrence of RMPP is not only determined by drug-resistance genes but may also be the result of a combination of factors.

Diagnostic Value of IP-10 Level in Plasma and Bronchoalveolar Lavage Fluid in Children with Tuberculosis and Other Lung Diseases.

Objectives: IP-10 has been proposed as a new diagnostic biomarker for Mycobacterium tuberculosis infection (MTBI). However, data on IP-10 concentration in bronchoalveolar lavage fluid (BALF) for pediatric tuberculosis are lacking. Aim: To determine IP-10 levels in unstimulated BALF and plasma in children with and without MTBI. Methods: IP-10 concentrations in BALF and plasma were measured in children hospitalized with suspected tuberculosis or other respiratory disease and scheduled for bronchoscopy. Thirty-five children were enrolled: 13 with suspected tuberculosis and 22 controls. The association between IP-10 and age was examined. Results: The IP-10 expression was increased in BALF compared to plasma (p = 0.008). We noticed higher BALF IP-10 levels in children with asthma, interstitial lung disease, and lung anomaly than in children with MTBI and other respiratory tract infections, but the differences were statistically insignificant. There was a moderate correlation between plasma and BALF IP-10 concentrations (rs = 0.46, p = 0.018). No correlation between IP-10 level and age was detected. Conclusions: IP-10 is detectable in unstimulated BALF in children with respiratory diseases, reaches higher concentrations in unstimulated BALF vs plasma, and does not correlate with age. However, it could not discriminate MTBI from other respiratory diseases.

Formation of Calprotectin-Derived Peptides in the Airways of Children with Cystic Fibrosis

Abstract Calprotectin is released by activated neutrophils along with myeloperoxidase (MPO) and proteases. It plays numerous roles in inflammation and infection, and is used as an inflammatory biomarker. However, calprotectin is readily oxidized by MPO-derived hypohalous acids to form covalent dimers of its S100A8 and S100A9 subunits. The dimers are susceptible to degradation by proteases. We show that detection of human calprotectin by ELISA declines markedly because of its oxidation by hypochlorous acid and subsequent degradation. Also, proteolysis liberates specific peptides from oxidized calprotectin that is present at inflammatory sites. We identified six calprotectin-derived peptides by mass spectrometry and detected them in the bronchoalveolar lavage fluid of children with cystic fibrosis (CF). We assessed the peptides as biomarkers of neutrophilic inflammation and infection. The content of the calprotectin peptide ILVI was related to calprotectin (r = 0.72, p = 0.01, n = 10). Four of the peptides were correlated with the concentration of MPO (r &amp;gt; 0.7, p ≤ 0.01, n = 21), while three were higher (p &amp;lt; 0.05) in neutrophil elastase–positive (n = 14) than –negative samples (n = 7). Also, five of the peptides were higher (p &amp;lt; 0.05) in the bronchoalveolar lavage fluid from children with CF with infections (n = 21) than from non-CF children without infections (n = 6). The specific peptides liberated from calprotectin will signal uncontrolled activity of proteases and MPO during inflammation. They may prove useful in tracking inflammation in respiratory diseases dominated by neutrophils, including coronavirus disease 2019.

CircZNF652 promotes the goblet cell metaplasia by targeting the miR-452-5p/JAK2 signaling pathway in allergic airway epithelia

Circular RNAs (circRNAs) play potentially important roles in various human diseases; however, their roles in the goblet cell metaplasia of asthma remain unknown. We sought to investigate the potential role and underlying mechanism of circZNF652 in the regulation of allergic airway epithelial remodeling. The differential expression profiles of circRNAs were analyzed by transcriptome microarray, and the effects and mechanisms underlying circZNF652-mediated goblet cell metaplasia were investigated by quantitative real-time PCR, RNA fluorescence in situ hybridization, Western blot, RNA pull-down, and RNA immunoprecipitation analyses. The roles of circZNF652 and miR-452-5p in allergic airway epithelial remodeling were explored in both the mouse model with allergic airway inflammation and children with asthma. One hundred sixty circRNAs were differentially expressed in bronchoalveolar lavage fluid of children with asthma versus children with foreign body aspiration, and 52 and 108 of them were significantly upregulated and downregulated, respectively. Among them, circZNF652 was predominantly expressed and robustly upregulated in airway epithelia of both the children with asthma and the mouse model with allergic airway inflammation. circZNF652 promoted the goblet cell metaplasia by functioning as a sponge of miR-452-5p, which released the Janus kinase 2 (JAK2) expression and subsequently activated JAK2/signal transducer and activator of transcription 6 (STAT6) signaling in the allergic airway epithelia. In addition, epithelial splicing regulatory protein 1, a splicing factor, accelerated the biogenesis of circZNF652 by binding to its flanking intron to promote the goblet cell metaplasia in allergic airway epithelial remodeling. Upregulation of circZNF652 expression in allergic bronchial epithelia contributed to the goblet cell metaplasia by activating the miR-452-5p/JAK2/STAT6 signaling pathway; thus, blockage of circZNF652 or agonism of miR-452-5p provided an alternative approach for the therapeutic intervention of epithelial remodeling in allergic airway inflammation.

Attenuated lncRNA NKILA Enhances the Secretory Function of Airway Epithelial Cells Stimulated by Mycoplasma pneumoniae via NF-κB.

The secretory function of airway epithelial cells is important in the pathogenesis of Mycoplasma pneumoniae pneumonia (MPP). To investigate the regulatory function of NKILA (nuclear factor-κB (NF-κB) interacting long noncoding RNA (lncRNA)) in MPP, we first detected NKILA as well as the concentration of interleukin 8 (IL-8) and tumor necrosis factor-α (TNF-α) in bronchoalveolar lavage fluid of children with MPP. Then, NKILA was knocked down in epithelial cells to investigate its effect on their secretory function. The results suggested that NKILA was downregulated in children with MPP, while IL-8 and TNF-α levels increased. Knockdown of NKILA in vitro promoted the inflammatory effects of Mycoplasma pneumoniae (MP) in epithelial A549 and BEAS-2B cells. Knockdown of NKILA promoted inhibitor of κBα (IκBα) phosphorylation and degradation, and NF-κB p65 nuclear translocation. Furthermore, RNA immunoprecipitation showed that NKILA could physically bind to IκBα in MP-treated A549 cells. Collectively, our data demonstrated that attenuation of NKILA enhances the effects of MP-stimulated secretory functions of epithelial cells via regulation of NF-κB signaling.

Broad range detection of viral and bacterial pathogens in bronchoalveolar lavage fluid of children to identify the cause of lower respiratory tract infections

BackgroundKnowledge on the etiology of LRTIs is essential for improvement of the clinical diagnosis and accurate treatment. Molecular detection methods were applied to identify a broad range of bacterial and viral pathogens in a large set of bronchial alveolar lavage (BAL) fluid samples. The patterns of detected pathogens were correlated to the clinical symptoms.MethodsBAL fluid samples and clinical data were collected from 573 hospitalized children between 1 month and 14 years of age with LRTIs, enrolled from January to December 2018. Pathogens were detected using standardized clinical diagnostics, with a sensitive, high-throughput GeXP-based multiplex PCR and with multiplex qPCR. Data were analyzed to describe the correlation between the severity of respiratory tract disease and the pathogens identified.ResultsThe pathogen detection rate with GeXP-based PCR and multiplex qPCR was significantly higher than by clinical routine diagnostics (76.09% VS 36.13%,χ2 = 8.191, P = 0.004). The most frequently detected pathogens in the BAL fluid were human adenovirus (HADV)(21.82%), Mycoplasma pneumoniae (20.24%), human rhinovirus (13.96%), Streptococcus pneumoniae (8.90%) and Haemophilus influenzae (8.90%). In 16.4% of the cases co-detection with two or three different pathogens was found. Viral detection rates declined with age, while atypical pathogen detection rates increased with age. Oxygen supply in the HADV and Influenza H1N1 infected patients was more frequent (49.43%) than in patients infected with other pathogens.ConclusionBroad range detection of viral and bacterial pathogens using molecular methods is a promising and implementable approach to improve clinical diagnosis and accurate treatment of LRTI in children.

Total and double-stranded DNA-specific immunoglobulin E in bronchoalveolar lavage fluid of children with human adenovirus pneumonia

BackgroundSome antibodies and autoreactive antibodies are associated with the severity of infectious diseases. The roles of humoral responses to lung inflammation in children with human adenovirus (HAdVs) pneumonia remain unknown. Patients and methodsA retrospective study was done to compare plasma immunoglobulin E (IgE) levels between HAdVs pneumonia patients and healthy children by searching the electronic medical record system of Guangzhou Women and Children's Medical Center. Then, a prospective study was performed for children with HAdVs pneumonia who needed flexible bronchoscopy for examination and treatment purposes during July 2017 to July 2019. We examined the IgE and autoreactive IgE levels in plasma and bronchoalveolar lavage fluid (BALF) of these children to explore their role in HAdVs pneumonia. ResultsA significantly higher level of IgE was found in plasma from children hospitalized with HAdVs pneumonia compared with that from healthy children in the same age range. Furthermore, the levels of IgE, double-stranded DNA (dsDNA), and double-stranded DNA-specific immunoglobulin E (dsDNA-IgE) in BALF were increased compared to plasma in children with HAdVs pneumonia. The levels of IgE, dsDNA, and dsDNA-IgE in BALF were significantly higher in the severe group compared to the non-severe group. The ability of IgE in BALF to recognize dsDNA was verified by the ELISPOT test. ConclusionsOur findings indicate that IgE and dsDNA-IgE in BALF may contribute to lung injury caused by HAdVs, especially in severe cases. Elevated dsDNA-IgE may serve as an indicator of severity in children with HAdVs pneumonia.

Study on the significance of cytokine in bronchoalveolar lavage fluid in children with refractory mycoplasma pneumoniae pneumonia and the correlation with serum C reactive protein

Objective To explore the levels and clinical significance of MCP-4, IL-25, TNF-α and CysLTR-1 in bronchoalveolar lavage fluid(BALF)of children with refractory mycoplasma pneumoniae pneumonia(RMPP)and their correlation with serum C-reactive protein(CRP). Methods One hundred and nine children diagnosed as RMPP who underwent fiberoptic bronchoscopy in acute stage(course of disease within 2 weeks)were selected as the experimental group.According to the manifestations of mucosa, secretions and lumen under bronchoscope, the patients were divided into RMPP1 group(68 cases of severe pathological injury under bronchoscope)and RMPP2 group(41 cases of mild pathological injury under bronchoscope). They were divided into RMPP1 wheezing group(20 cases), RMPP1 non- wheezing group(48 cases), RMPP2 wheezing group(15 cases)and RMPP2 non-wheezing group(26 cases).15 children with non-mycoplasma pneumoniae pneumonia(NMPP)and non-wheezing lobar pneumonia in the same period were selected as control group 1.At the same time, 15 children without pneumonia underwent bronchial foreign body(FB)removal as control group 2.The levels of MCP-4, IL-25, TNF-α and CysLTR-1 in BALF of children in experimental group were determined by double antibody sandwich ELISA.Serum CRP, D dimer(DD), ALT and peripheral blood neutrophil percentage(N%)were also detected. Results (1)The levels of CRP, DD, ALT and N% in RMPP1 group with severe bronchoscopic manifestations were higher than those in RMPP2 group with relatively mild bronchoscopic manifestations(all P 0.05). (3)CRP was positively correlated with IL-25, MCP-4 and TNF-a(all P<0.05), but not with CysLTR-1. Conclusion (1)Clinical laboratory indicators such as CRP, DD, ALT and N% can assist in early identification of RMPP.The higher the above indicators, the more serious performance of RMPP under microscope.(2)Cytokines MCP-4, IL-25, CysLTR-1 and TNF-α all participate in the pathogenesis of RMPP, and may play an important role in the occurrence of wheezing and development of asthma in children induced by MP infection.(3)Serum CRP levels were positively correlated with the levels of IL-25, MCP-4 and TNF-α in BALF of RMPP wheezing children.Both MCP-4 and IL-25 selectively affected Th2-induced Th2 cells.CRP is associated with IL-25 and MCP-4 who mediated immune inflammation injury.It is speculated that CRP may also cause Th2-mediated immune inflammation injury by affecting Th2 cells. Key words: Refractory mycoplasma pneumoniae pneumonia; Electronic fiberoptic bronchoscope; Cell factor; C-reactive protein; Children

Multiplex PCR of bronchoalveolar lavage fluid in children enhances the rate of pathogen detection

BackgroundCulturing of bronchoalveolar lavage (BAL) fluid is a commonly used method for pathogen detection in pneumonia. However, the sensitivity is low, especially in patients pre-treated with anti-infective agents. The early detection of a pathogen is crucial for the outcome of respiratory tract infections. For bloodstream infections, a multiplex polymerase chain reaction (PCR) assay (SeptiFast®, SF) is available for improved pathogen detection from blood.ObjectiveThe aim of the present study was to determine whether the SF assay is applicable to the BAL of children with pulmonary infections and whether the frequency of pathogen detection is enhanced by the use of this multiplex PCR method.MethodsWe investigated 70 BAL samples of 70 children simultaneously by culture and multiplex PCR. The frequency of pathogen detection was compared.ResultsPathogens were detected more frequently by SF than by culture (83% vs. 31%; p < 0.001). This advantage was shown for immunocompetent patients (p = 0.001) as well as for immunocompromised patients (p = 0.003). The majority (38/44; 86%) of the Gram positive cocci were only detected by SF. Fungal organisms were detected in 7/70 patients (10%) by SF and in 2/70 (3%) by culture (p = 0.125).ConclusionCompared to conventional culture, the use of the SF assay on the BAL of children with pneumonia increases pathogen detection rates and therefore adds important information to guide anti-infective therapy.

Detecting respiratory viruses in children with protracted bacterial bronchitis.

This study is to investigate the status and clinical significance of respiratory viruses in bronchoalveolar lavage fluid (BALF) of children with PBB. Sixty-eight children with PBB aged from 3 months to 5 years were enrolled and retrospectively reviewed from January 2014 to December 2017. Thirty-five children with persistent pneumonia or chronic pneumonia were matched as controls. Bronchoalveolar lavage fluid samples were collected for respiratory virus detection and bacterial culture. The detection rate of bacteria in BALF of children with PBB was 61.8%, which was significantly higher than that of control group (20%) (P < 0.001). The detection rate of virus in BALF of children with PBB was 23.5%, including 6 (8.8%) of rhinovirus, 4 (5.9%) of parainfluenza virus type 3, 2(2.9%) of bocavirus, 2 (2.9%) of respiratory syncytial virus 1 (1.5%) of human metapneumonia virus and 1 (1.5%) of influenza virus A. 10 cases (28.6%) of virus were detected in the control group, including 3 (8.6%) respiratory syncytial virus, 3 (8.6%) rhinovirus and 2 (5.7%) bocavirus. There was no significant difference of viral detection rate between the two groups (P = 0.577). Respiratory viruses can be detected in BALF of children with PBB, However, there is no evidence that PBB is virus-induced.

Diagnostic Yield of Xpert MTB/RIF in Bronchoalveolar Lavage in Children with Probable Pulmonary Tuberculosis

To evaluate utility of Xpert MTB/RIF in bronchoalveolar lavage fluid in children with probable pulmonary tuberculosis. Children with probable pulmonary tuberculosis with negative smear and Xpert on induced sputum/gastric aspirate were subjected to bronchoalveolar lavage (BAL) for Xpert assay and mycobacterial liquid culture. Data of children <14 y undergoing bronchoscopy for suspected MDR-TB (n=12) were also analyzed. The sensitivity of Xpert in BAL fluid for diagnosis of probable and confirmed pulmonary tuberculosis was calculated with clinico-radiological diagnosis and culture as gold standards, respectively. Of 41 enrolled children, 24 (58.5%) had Xpert positive in BAL fluid and 11 (26.8%) had culture confirmed tuberculosis (BAL fluid;10; sputum,1). The sensitivity of Xpert in BAL fluid among probable and culture confirmed tuberculosis cases was 58.5% (24/41) and 81.8% (9/11), respectively. Xpert in bronchoalveolar lavage fluid has good sensitivity in both probable and confirmed pulmonary tuberculosis in children.

Associations between patient clinical characteristics and the presence of cytomegalovirus DNA in the bronchoalveolar lavage fluid of children with recurrent wheezing

BackgroundThis study aimed to investigate the occurrence of Cytomegalovirus (CMV) DNA in the Bronchoalveolar lavage fluid (BALF) of children with recurrent wheezing and to identify associations with certain patient clinical characteristics.MethodsIn this cross-sectional study, pediatric patients (age < 36 months) admitted to Soochow University Hospital with recurrent wheezing (≥ 4 episodes of wheezing per year) were enrolled in the study. Cytomegalovirus DNA from their BALF was detected by real-time PCR. Subpopulations of blood immunoglobulins and T lymphocytes were quantified. The clinical characteristics of patients with and without BALF CMV DNA were compared. Comparisons of non-normally distributed continuous variables between groups were made using the Mann-Whitney U-test. Comparisons of frequency distributions were made using the Chi-squared test. Spearman’s rank correlation coefficient was used to evaluate correlations between the number of CMV DNA copies and continuous variables.ResultsA total of 111 patients aged 4 to 36 months (median 14.0 (IQR 8.0–22.0) months) were enrolled on to the study. Cytomegalovirus DNA was detected in 51.4% of patients (n = 111) with recurrent wheeze and was more prevalent among those aged 12 to 36 months with a positive modified asthma predictive index (mAPI) (n = 38, median 23.5 (IQR 19.7–31.2) months) than in those of the same age group with a negative mAPI (n = 25, median 15.0 (IQR 13.0–19.0) months) (57.9% vs. 20.0%, p = 0.003). Bronchoalveolar lavage fluid CMV DNA copy number [median 7560 (IQR 1200–71,150) copies/mL] was positively correlated with the duration of hospitalization (r = 0.33, p = 0.013), and negatively correlated with patient age (r = − 0.41, p = 0.002) and the percentage of BALF eosinophils (r = − 0.38, p = 0.004).ConclusionsCMV infection or reactivation in children with recurrent wheeze is associated with certain clinical characteristics, including younger age and lower levels of BALF eosinophils. Higher CMV DNA copy numbers were associated with a longer duration of hospitalization. Further studies are needed to address whether specific antiviral treatment could be beneficial for BALF CMV positive patients.

The role of granulocyte macrophage colony stimulating factor in hospitalized children with Mycoplasma pneumoniae pneumonia.

Inappropriate inflammatory response in children with M. pneumoniae infection might be associated with disease severity. The role of Granulocyte macrophage colony stimulating factor (GM-CSF) in hospitalized children with Mycoplasma pneumoniae pneumonia (MPP) has not been fully discussed. Clinical and laboratory data of a total 40 children with MPP were collected. GM-CSF and myeloperoxidase (MPO) were detected by ELISAs. Meanwhile, normal human bronchial epithelium was infected by M.pneumoniae and neutrophils were stimulated by GM-CSF to explore GM-CSF and MPO release in supernatant, respectively. Compared to control group, a significant increased percentage of neutrophils and decreased percentage of macrophages in bronchoalveolar lavage fluid of children with MPP was observed (P<0.05). Children with MPP had significantly higher levels of GM-CSF (P=0.0047) and MPO (P=0.0002) in BALF compared to the controls. Level of GM-CSF in BALF was associated with duration of fever (r=0.42, P=0.007) and strongly correlated with level of MPO (r=0.075, P=0.0005). Levels of GM-CSF and MPO significantly decreased (both P<0.05) after treatment. Invitro, M.pneumoniae induced GM-CSF expression in a time-dependent manner during a 72-h period (P<0.05) and MPO secretion significantly increased by recombinant human GM-CSF stimulation at 24h (P<0.05). GM-CSF could be induced by M.pneumoniae infection in vivo and vitro. Childen with high level GM-CSF had longer duration of fever. GM-CSF probably plays a vital role in neutrophil inflammation in M.pneumoniae infection.

The clinical significance of T-cell subset, cytokine levels in bronchoalveolar lavage fluid of children with mycoplasma pneumoniae pneumonia

Objective To explore the levels and clinical significances of T-cell subset, interleukin(IL)-6, interferon(IFN)-γ, and monocyte chemotactic protein(MCP)-1 in bronchoalveolar lavage fluid(BALF) of children with mycoplasma pneumoniae pneumonia(MPP). Methods Thirty-six children with pneumonia were admitted in our hospital from October 2014 to March 2016.They were divided into refractory mycoplasma pneumoniae pneumonia (RMPP) group (n=18) and MPP group(n=18). Sixteen cases with bronchial foreign body were selected as the control group.The levels of IL-6, MCP-1, IFN-γ and T-cell subset in BALF were detected by ELISA and alkaline phosphatase-anti-alkaline phosphatase(APAAP) bridging enzyme linked immunosorbent assay. Results (1)The levels of CD3+ and CD8+ T cells in BALF of MPP group and RMPP group were higher than those in the control group (P 0.05). (2) The level of IL-6 in acute phase of RMPP group was higher than that in the control group (P<0.05). The levels of IL-6 in acute phase of RMPP group and MPP group were both higher than that in recovery period (P<0.05, respectively). (3)The levels of IFN-γ in the acute phase of RMPP group and MPP group were higher than that in the control group, and the differences were statistically significant (P<0.05). The IFN-γ levels in acute and recovery phases of RMPP group were both significantly higher than those in MPP group respectively(P< 0.05). (4)The level of MCP-1 in BALF in control group was lower than those in the acute phase of RMPP group and MPP group(P<0.05). The level of MCP-1 in RMPP acute group was significantly higher than that in MPP acute group, and higher in acute phase than those in recovery phase both in two groups(P<0.05). Conclusion There are cell immune regulation and function disorders when children are infected mycoplasma pneumoniae.IL-6, IFN-γ and MCP-1 may participate in the pathogenesis of MPP, and more importantly, IFN-γ and MCP-1 may be the important indicator to forecast the severity of MPP. Key words: Mycoplasma pnuemoniae; Bronchoalveolar lavage fluid; T-cell subset; Cytokine,

Expressions of granulocyte-macrophage colony-stimulating factor and myeloperoxidase in bronchoalveolar lavage fluid in children with Mycoplasma pneumoniae pneumonia and its clinical significance

目的 检测肺炎支原体肺炎(MPP)患儿支气管肺泡灌洗液(BALF)中粒细胞巨噬细胞集落刺激因子(GM－CSF)表达水平，探讨其在MPP中的临床意义。 方法 收集2013年1月至2014年12月在苏州大学附属儿童医院呼吸科住院的40例MPP患儿BALF标本，30例患儿因病情治疗需要行第2次支气管肺泡灌洗，收集治疗后(恢复期)BALF标本，并采用酶联免疫吸附法检测GM－CSF和髓过氧化物酶(MPO)水平。同时收集患儿临床和实验室检查资料。以15例急性气管支气管异物患儿的BALF作为对照。 结果 MPP患儿BALF中性粒细胞分类高于对照组[(62.1±23.5)%比(4.5±3.3)%，t＝2.73，P<0.01]。肺炎支原体DNA高值组(≥ 1010 copies/L)白细胞总数[(9.7±4.2)×109/L比(6.4±2.2)×109/L，t＝2.131，P＝0.04]和中性粒细胞数[(6.9±3.4)×109/L比(3.6±1.2)×109/L，t＝2.599，P＝0.01]均高于肺炎支原体DNA低值组(<1010 copies/L)。MPP患儿BALF中GM－CSF[(153.8±36.4) ng/L比(23.0±25.7) ng/L, P<0.05]和MPO[(21.4±7.0) ng/L比(13.8±3.5)ng/L，P<0.05]水平均明显高于对照组。MPP患儿BALF中GM－CSF水平与热程(r＝0.42，P<0.05)及MPO水平均呈正相关(r＝0.75, P<0.05)。恢复期GM－CSF和MPO水平均明显降低(均P<0.05)。 结论 GM－CSF可能在儿童MPP发生发展中起重要作用，可作为疾病转归的有效预测指标之一。