This paper was mainly to discuss the potential role and mechanism of Lianhua Qingwen Capsules(LHQW) in inhibiting pathological inflammation in the model of acute lung injury caused by bacterial infection. For in vitro study, the mRNA expression of MCP-1 in RAW264.7 cells and THP-1 cells, the content of MCP-1 in cell supernatant, as well as the effect of LHQW on chemotaxis of macrophages were detected. For in vivo study, mice were randomly divided into 7 groups, including normal group, model group(LPS 5 mg·kg~(-1)), LHQW 300, 600 and 1 200 mg·kg~(-1)(low, middle and high dose) groups, dexamethasone 5 mg·kg~(-1) group and penicillin-streptomycin group. Then, the anal temperature was detected two hours later. Dry weight and wet weight of lung tissues in mice were determined; TNF-α and MCP-1 levels in alveolar lavage fluid and MCP-1 in serum were detected. In addition, the infiltration of alveolar macrophages was also observed and the infiltration count of alveolar macrophages was measured by CCK-8 method. HE staining was also used to observe the inflammatory infiltration of lung tissues in mice. Both of the in vitro and in vivo data consistently have confirmed that: by down-regulating the expression of MCP-1, LHWQ could efficiently decrease the chemotaxis of monocytes toward the pulmonary infection foci, thus blocking the disease development in ALI animal model.
Acute Lung Injury ; microbiology ; Animals ; Bacterial Infections ; drug therapy ; Bronchoalveolar Lavage Fluid ; Capsules ; Chemokine CCL2 ; metabolism ; Chemotaxis ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Lipopolysaccharides ; Lung ; Macrophages ; drug effects ; Mice ; RAW 264.7 Cells ; Random Allocation ; THP-1 Cells ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo analyze the clinical characteristics of protracted bacterial bronchitis (PBB) in children.

METHODThe clinical data of patients seen from October, 2010 to March, 2014 in Department of Respiratory Diseases of our hospital were retrospectively analyzed. Inclusion criteria were over 4 weeks cough, receiving fiberoptic bronchoscopy, positive bacterial culture and (or) the increased percentage of neutral granulocytes in bronchoalveolar lavage fluid (BALF).

RESULTTwenty eight patients were involved, 26 were male (93%) and two were female (7%). The median age of patients was 8.5 months. The median duration of cough was four weeks. The average length of hospital stay was (8.3 ± 3.9)days. The main clinical feature was wet cough in 28 cases, wet cough with wheezing was seen in 21 cases. The wet cough phase distribution was irregular in 21 cases. The crackles with wheeze (in 21 cases) was main signs of PBB. The percentage of CD3⁻ CD16⁺ 56⁺ cells increased in peripheral blood. The fiberoptic bronchoscopic manifestations of PBB were luminal mucosal edema. Eleven patients also had airway malacia. The neutrophil median in BALF was 0.2. The positive rate of bacterial culture of BALF was 36%. The main bacteria were Streptococcus pneumoniae (50%) and Haemophilus influenzae (30%). The main treatment for PBB patients included amoxycillin/clavulanate potassium and second-generation cephalosporins. The average duration of treatment was (17.3 ± 3.2)days, the prognosis was good.

CONCLUSIONPBB is common in male infants. Persistent wet cough with wheezing was the main characteristic of PBB. PBB is commonly accompanied by immune dysfunction and airway malacia, and the pathogens were Streptococcus pneumoniae and Haemophilus influenzae.

Bacterial Infections ; drug therapy ; pathology ; Bronchitis ; drug therapy ; microbiology ; pathology ; Bronchoalveolar Lavage Fluid ; Bronchoscopy ; Cough ; Female ; Haemophilus influenzae ; isolation & purification ; Humans ; Infant ; Male ; Respiratory Sounds ; Retrospective Studies ; Streptococcus pneumoniae ; isolation & purification

OBJECTIVENoncystic 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 SOURCESWe 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 SELECTIONWe 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.

RESULTSThe 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 H. 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. aureus, 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.

CONCLUSIONSH. 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.

Bronchiectasis ; microbiology ; Bronchoalveolar Lavage Fluid ; microbiology ; Haemophilus influenzae ; pathogenicity ; Humans ; Pseudomonas aeruginosa ; pathogenicity ; Sputum ; microbiology

OBJECTIVETo compare the detection rates of Mycoplasma pneumoniae (MP) from nasopharyngeal aspirates (NPA) and bronchoalveolar lavage fluid (BALF) in children with pneumonia.

METHODSA total of 164 hospitalized children with pneumonia were enrolled. NPA and BALF of these children were collected within 24 hours of admission, and MP-DNA was detected by fluorescence quantitative PCR. Venous blood samples of all these children were collected within 24 hours of admission and on days 7-10 of treatment, and serum MP-IgM was detected using ELISA.

RESULTSThe positive rate of MP-DNA in NAP of the 164 cases was 51.8% , which was lower than 63.4% as the detection rate of MP-IgM in serum (P=0.044), and the two detection rates were moderately consistent with each other (Kappa=0.618, P<0.01). The positive rate of MP in BALF was 71.3%, which was not significantly different with that of MP-IgM in serum (P>0.05), and the detection rates were well consistent (Kappa=0.793, P<0.01). The detection rate of MP in NPA was lower than that in BALF (P<0.01), with moderate consistency between two of them (Kappa=0.529, P<0.01). The median MP copy number in BALF was significantly higher than that in NPA (P<0.01). The MP detection rates in NPA and BALF were significantly different among different courses of disease (P<0.05). As the course of disease extended, the MP detection rates in both NPA and BALF showed a declining trend; children with MP pneumonia of 1-2 weeks' duration and 2-4 weeks' duration had a higher MP-DNA detection rate in BALF than in NPA (P<0.05).

CONCLUSIONSMP-DNA in BALF has a high sensitivity, with a great significance for early diagnosis of MP pneumonia, while NPA MP-DNA tests may lead to a missed diagnosis.

Adolescent ; Bronchoalveolar Lavage Fluid ; microbiology ; Child ; Child, Preschool ; DNA, Bacterial ; analysis ; Female ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Pneumonia, Mycoplasma ; diagnosis

Lung disease caused by nontuberculous mycobacteria (NTM) represents an increasing proportion of all mycobacterial diseases. We investigated recent occurrences of NTM and evaluated the clinical significance of NTM isolates from 752 respiratory specimens collected from patients at National Health Insurance Service Ilsan Hospital between January 2007 and May 2011. Specimens were incubated on solid and liquid media (BACTEC MGIT 960, BD, USA) for 6-8 weeks, and PCR and reverse blot hybridization were performed (REBA Myco-ID, Molecules & Diagnostics, Korea). Clinical features of the patients were reviewed through medical records. The most frequently isolated organism was Mycobacterium avium (46.7%), followed by M. intracellulare (14.8%), M. fortuitum (7.2%), and M. abscessus (6.6%). The most common mycobacteria among definitive cases of NTM lung disease were M. avium (42/351, 12.0%), M. intracellulare (19/111, 17.1%), M. abscessus (11/50, 22.0%), M. massiliense (4/13, 30.8%), and M. fortuitum (4/54, 7.4%). Clinically significant cases of NTM lung disease increased from 4 patients in 2007 to 32 in 2011. The mean patient age was 64 yr (range: 35-88 yr), and 58 (64%) patients were women. Patients suffered from cough, productive sputum, and hemoptysis. In summary, the most common mycobacteria causing NTM lung disease were M. avium and M. intracellulare; however, cases of M. massiliense and M. abscessus infection are on the rise in Korea.
Adult ; Aged ; Aged, 80 and over ; Bronchoalveolar Lavage Fluid/microbiology ; DNA, Bacterial/analysis ; Female ; Hospitals, General/standards/*trends ; Humans ; Lung Diseases/diagnosis/*microbiology ; Male ; Middle Aged ; Mycobacterium Infections, Nontuberculous/diagnosis/*microbiology ; Nontuberculous Mycobacteria/genetics/*physiology ; Nucleic Acid Hybridization ; Polymerase Chain Reaction ; Reagent Kits, Diagnostic ; Republic of Korea ; Sputum/microbiology

Accurate quantification of mycobacterial load is important to evaluate disease severity and to monitor the course of treatment in tuberculosis (TB). We evaluated the quantitative capability of the AdvanSure TB/NTM real-time PCR kit (LG Life Science, Korea) to determine the cycle threshold (Ct) for mycobacterial burden. We retrospectively analyzed data from 108 patients whose respiratory specimens (sputums and bronchoalveolar lavage fluids) were positive for Mycobacterium tuberculosis complex (85 culture-positive and 23 culture-negative specimens). We compared Ct values with grades of acid-fast bacilli (AFB) staining, semi-quantitative colony count on solid medium, and time to positivity (TTP) in liquid and solid media. We also investigated the cutoff Ct value for predicting stain-positive status. Ct value showed significant reverse correlation with AFB staining grade (r(s)=-0.635, P<0.01). Ct value significantly decreased as the semi-quantitative counts on the solid medium increased (P<0.001), and the mean Ct value of each of the groups 1+, 2+, 3+, and 4+ were 29.0, 30.0, 27.1, and 25.5, respectively. A weak correlation between Ct value and TTP in liquid and solid media was observed (r(s)=0.468 and 0.365, respectively). A cutoff Ct value of <33.2 best predicted stain positivity, with a sensitivity of 95.0% and a specificity of 32.0%. Our findings suggest the potential use of AdvanSure TB/NTM real-time PCR kit for quantitatively determining bacterial burden, albeit with some enhancements.
Area Under Curve ; Bronchoalveolar Lavage Fluid/microbiology ; DNA, Bacterial/*analysis ; Humans ; Mycobacterium tuberculosis/genetics/*isolation & purification ; Phenotype ; ROC Curve ; Reagent Kits, Diagnostic ; *Real-Time Polymerase Chain Reaction ; Retrospective Studies ; Sensitivity and Specificity ; Sputum/microbiology ; Tuberculosis/diagnosis/microbiology

OBJECTIVETo analyze the clinical characteristics of Mycoplasma pneumoniae-associated hemophagocytic syndrome (MP-HLH).

METHODA retrospective investigation of the clinical manifestation, laboratory test, imagelogy, clinical course and outcome of 3 cases with MP-HLH seen between June 2013 and July 2013 in Shenzhen Children's Hospital, and review of relevant literature were conducted.

RESULTOf the 3 cases of MP-HLH, 2 were males, one was female, the ages were 1 year, 3 years and 6 years, respectively. They had no underlying disease previously. All the 3 cases had onset of fever, cough as main symptoms. Diagnosis of refractory Mycoplasma pneumoniae pneumonia was made, which was accompanied by decreased neutrophils [(0.08-0.68)×10(9)/L], hemoglobin [(79-103) g/L], platelet [(64-157)×10(9)/L], plasma fibrinogen [(1.3-1.5) g/L], lactate dehydrogenase [(1,170-1,285) U/L] and increased serum ferritin [(936.7-39 789.0) µg/L] in the third week of course. In two cases the T lymphocytes decreased, and the NK cell activity decreased significantly in one. Bone marrow cytology showed prompted bone marrow hyperplasia, and the phenomenon of phagocytosed blood cells. CT scan was performed for all the cases and consolidation with pleural effusion were shown. Two cases were admitted to PICU, and required endotracheal intubation and mechanical ventilation. Flexible bronchoscopy and bronchial lavage were performed and bronchial cast was found in two cases. All of them were treated with macrolide combined with other antibiotics, glucocorticoids and gamma globulin combination therapy, including one case given dexamethasone [10 mg/(m2·d)], cyclosporine[6 mg/(kg·d)], etoposide [150 mg/(m2·d)] chemotherapy. Two cases were cured, and 1 case died. The authors summarized the 18 cases reported in domestic and foreign literature. Foreign children were diagnosed and treated with steroids in 1-2 weeks, and 10 cases were cured, and 2 cases died. They died of massive hemorrhage and meningoencephalitis, and domestic children were diagnosed and treated within two to 4 weeks after onset, 5 cases were cured, one case died of severe pneumonia.

CONCLUSIONMP-HLH is a rare disease in children, and had acute onset, rapid progression and high mortality. Early treatment with steroids was associated with a good prognosis, the key to successful treatment is early diagnosis and treatment, avoiding the immune cascade. Too late a diagnosis or development of serious complications may lead to death.

Anti-Bacterial Agents ; therapeutic use ; Bronchoalveolar Lavage Fluid ; Bronchoscopy ; Child ; Child, Preschool ; Fatal Outcome ; Female ; Fever ; Glucocorticoids ; therapeutic use ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; drug therapy ; microbiology ; Macrolides ; therapeutic use ; Male ; Mycoplasma pneumoniae ; isolation & purification ; Pleural Effusion ; Pneumonia, Mycoplasma ; complications ; diagnosis ; drug therapy ; Respiration, Artificial ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

BACKGROUNDThe 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.

METHODSThis 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 PIC 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.

RESULTSThere 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.

CONCLUSIONSPIC>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.

Aged ; Bronchoalveolar Lavage Fluid ; microbiology ; Female ; Humans ; Male ; Middle Aged ; Pneumonia, Ventilator-Associated ; diagnosis ; Prospective Studies

Bronchiectasis is a chronic lung disorder and a number of bacterial pathogens are involved. However, 30%-40% of sputum and purulent samples in good quality failed to grow any pathogenic bacteria, making it difficult to confirm the pathogen. In this study, we collected bronchoalveolar lavage fluid from a bronchiectasis patient undergoing acute exacerbation, and sent for 16S rDNA pyrosequencing by a 454 GS Junior machine. Metagenomic analysis showed the composition of bacterial community in sample was complex. More than a half of reads (51.3%) were from Pseudomonas aeruginosa. This result was corresponding with the culture result but came out 2 d earlier, which is meaningful for early diagnosis and treatment. The detection with 16S rDNA pyrosequencing technology is more sensitive and rapid than routine culture, and can detect the co-infection or symbiosis in airway, giving us a novel and convenient approach to perform rapid diagnosis.
Bronchiectasis ; microbiology ; Bronchoalveolar Lavage Fluid ; chemistry ; microbiology ; Early Diagnosis ; Female ; Humans ; Metagenome ; genetics ; Metagenomics ; methods ; Middle Aged ; Pseudomonas Infections ; microbiology ; Pseudomonas aeruginosa ; genetics ; isolation & purification ; RNA, Ribosomal, 16S ; genetics ; Time Factors

Rothia mucilaginosa is a gram-positive coccus of the family Micrococcaceae. R. mucilaginosa is considered a part of the normal flora of the human oropharynx and upper respiratory tract and lower respiratory tract infections attributable to R. mucilaginosa are not frequent. We present a case of pneumonia, in which the R. mucilaginosa infection was diagnosed by quantitative cultures of a bronchoalveolar lavage (BAL) specimen. A 46-yr-old woman with B lymphoblastic lymphoma was admitted to the hospital for scheduled chemotherapy. Her chest computed tomography (CT) scan revealed bilateral multifocal nodular and patchy consolidation in both lungs. Investigation of the BAL specimen revealed that 7% of leukocytes had intracellular gram-positive cocci. The quantitative cultures of the BAL specimen grew mucoid, non-hemolytic, and grayish convex colonies on blood agar at a count of approximately 200,000 colony-forming units/mL. The colonies were identified as R. mucilaginosa. The patient was empirically treated with levofloxacin for 7 days, after which findings on the chest radiograph and CT scan improved. She was discharged with improvement on hospital day 46. To our knowledge, this is the first report of R. mucilaginosa pneumonia diagnosed in Korea. Quantitative culture of BAL specimen and examination of intracellular organisms are crucial for assessing the clinical significance of R. mucilaginosa recovered from the lower respiratory tract.
Bronchoalveolar Lavage Fluid/*microbiology ; Female ; Humans ; Lung/radiography ; Lymphoma/complications/*diagnosis ; Micrococcaceae/*isolation & purification ; Middle Aged ; Pneumonia/complications/*diagnosis/microbiology ; Tomography, X-Ray Computed