Airway microbiota correlated with pulmonary exacerbation in primary ciliary dyskinesia patients.

IMPORTANCE: PCD is a rare disease characterized by productive cough, rhinitis, and recurrent infections of the upper and lower airways. Because the diagnosis of PCD is often delayed, patients receive more antibiotics, experience a heavier financial burden, and have a worse prognosis; thus, it is very important to identify the pathogeny and use the correct antibiotic. In this large single-center study of PCD microbiota, we identified an outline of the bacterial microbes from the respiratory tract; furthermore, we found that the microbiota diversity in pediatric sputum was richer than that in pediatric BALF through sequencing, indicating a heterogeneous community structure. The microbiota diversity and richness were lower during pulmonary exacerbation than during pulmonary stabilization. A significantly higher abundance of Pseudomonas had a moderate distinguishing effect for lung exacerbation, which attracted more attention for the study of Pseudomonas therapy in pediatric patients with PCD.

[Congenital pulmonary alveolar proteinosis in a neonate]. / æ–°ç”Ÿå„¿å ˆå¤©æ€§è‚ºæ³¡è›‹ç™½æ²‰ç§¯ç—‡1例.

The male patient was referred to the hospital at 44 days old due to dyspnea after birth and inability to wean off oxygen. His brother died three days after birth due to respiratory failure. The main symptoms observed were respiratory failure, dyspnea, and hypoxemia. A chest CT scan revealed characteristic reduced opacity in both lungs with a "crazy-paving" appearance. The bronchoalveolar lavage fluid (BALF) showed periodic acid-Schiff positive proteinaceous deposits. Genetic testing indicated a compound heterozygous mutation in the ABCA3 gene. The diagnosis for the infant was congenital pulmonary alveolar proteinosis (PAP). Congenital PAP is a significant cause of challenging-to-treat respiratory failure in full-term infants. Therefore, congenital PAP should be considered in infants experiencing persistently difficult-to-treat dyspnea shortly after birth. Early utilization of chest CT scans, BALF pathological examination, and genetic testing may aid in early diagnosis.

Case report: A study on the de novo KMT2D variant of Kabuki syndrome with Goodpasture's syndrome by whole exome sequencing.

Kabuki syndrome (KS) is a rare genetic disorder characterized by dysmorphic facial features, skeletal abnormalities, and intellectual disability. KMT2D and KDM6A were identified as the main causative genes. To our knowledge, there exist no cases of KS, which were reported with pneumorrhagia. In this study, a 10-month-old male was diagnosed to have KS with typical facial features, skeletal anomalies, and serious postnatal growth retardation. Whole exome sequencing of the trio family revealed the presence of a de novo KMT2D missense variant (c.15143G > A, p. R5048H). The child was presented to the pediatric emergency department several times because of cough, hypoxemia, and anemia. After performing chest CT and fiberoptic bronchoscopy, we found that the child had a pulmonary hemorrhage. During research on the cause of pulmonary hemorrhage, the patient's anti-GBM antibodies gradually became positive, and the urine microalbumin level was elevated at the age of 12-month-old. After glucocorticoids and immunosuppressant therapy, the patient became much better. But he had recurrent pulmonary hemorrhage at the age of 16 months. Therefore, the patient underwent digital subtraction angiography (DSA). However, the DSA showed three abnormal bronchial arteries. This single case expands the phenotypes of patients with KS and Goodpasture's syndrome, which were found to have a de novo KMT2D missense variant.

Characteristics of Lung Microbiota in Children's Refractory Mycoplasma pneumoniae Pneumonia Coinfected with Human Adenovirus B.

BACKGROUND: Both M. pneumoniae and human adenovirus (HAdV) are common causative agents of lower respiratory tract infection in children; nonetheless, the lung microbiota in patients with coinfection of HAdV and M. pneumoniae remain unexplored. METHODS: Thirty-two children, diagnosed with refractory M. pneumoniae pneumonia (RMPP), entered into the one-year study from July 1, 2019 to June 30, 2020. Among them, twenty-one entered into the M. pneumoniae monoinfection (MP) group and eleven entered into the M. pneumoniae and HAdV coinfection (MP&ADV) group. The characteristics of the clinical findings were examined, and the lung microbiota was analyzed by metagenomic next generation sequencing (mNGS). RESULTS: Eleven patients in the MP&ADV group were coinfected with human mastadenovirus species B. The fever days lasted for significantly longer periods in the MP&ADV group than in the MP group (P < 0.05). The percentage of CD16+CD56+ cells was significantly higher in the MP&ADV group than that in the MP group (P < 0.05). There were no significant differences in α-diversity between the MP and MP&ADV groups, but the ß-diversity was clearly higher in the MP&ADV group than that in the MP group (P < 0.05). At the microbial level, the top phylum of the MP BALF microbiota was Tenericutes; in contrast, it was Preplasmiviricota in the MP&ADV BALF. There were significant differences in the relative abundance of Tenericutes and Preplasmiviricota between the two groups (P < 0.001). There was a strong positive correlation between human mastadenovirus B and fever days, M. pneumoniae and level of IgA, and a strong negative correlation between Mycoplasma pneumoniae and PCT. CONCLUSIONS: In RMPP, the BALF microbiota in children with mono M. pneumoniae infection was simpler than those with coinfection with human mastadenovirus B. Prolonged fever days were associated with human mastadenovirus B coinfection.

Clinical features and risk factors for severe influenza in children: A study from multiple hospitals in Shanghai.

BACKGROUND: The incidence and mortality of influenza in children had risen, but data are limited on children with severe influenza virus infection in China. METHODS: We conducted a retrospective case-control study and collected the patients' clinical data. Clinical data including demography, clinical presentation, laboratory findings, radiologic findings, treatment and outcomes were collected. Children were clinically confirmed to have virus infection in Shanghai in three hospitals from June 2014 to June 2019. RESULTS: During the study, 36,047 children were enrolled. Among them, 118 met the criteria for severe flu. Clinical symptoms such as fever, cough, gastrointestinal symptoms, coma and epilepsy were higher in the severe group. Complications such as pneumorrhagia, heart failure, septic shock, acute renal failure and influenza-associated encephalitis were higher in the severe influenza group than the death group. The laboratory findings including decreased hemoglobin, high alanine aminotransferase, high urea nitrogen and high lactate levels were risk factors for death in children with influenza. CONCLUSION: Influenza-associated encephalopathy (IAE), acute respiratory distress syndrome (ARDS) were the common clinical manifestations and complications for the severe influenza, and delayed use of oseltamivir was found to be associated with fatality.

Highly homogeneous microbial communities dominated by Mycoplasma pneumoniae instead of increased resistance to macrolide antibiotics is the characteristic of lower respiratory tract microbiome of children with refractory Mycoplasma pneumoniae pneumonia.

BACKGROUND: Although researchers have found that the microbiota changed during the lower respiratory tract (LRT) infection, little was known about the association between LRT microbiome and refractory M. pneumoniae pneumonia (RMPP). METHODS: From June 28th, 2019 to March 23rd, 2020, we enrolled fifty-two children diagnosed with RMPP or non-refractory M. pneumoniae pneumonia (NRMPP), and characterized the structure and function of microbiota in the bronchoalveolar lavage fluid (BALF) by metagenomic next generation sequencing (mNGS). RESULTS: Based on Bray-Curtis distance between samples, samples in RMPP group were highly homogeneous, and Shannon index in the RMPP group was much lower than NRMPP group while Simpson index, which presents the degree of dominance, was higher in RMPP group. The dominant taxon with relative abundance greater than 50% was merely Mycoplasma among RMPP and NRMPP patients, but the proportions of other taxonomic distribution were different. M. pneumoniae was the dominant species and occupied almost all niches in the vast majority of RMPP patients, whereas the other genera were dramatically lower. The NRMPP group was more enriched in antibiotic resistance genes (ARGs) than the RMPP group, and also exhibited a greater relative abundance of macrolide antibiotics resistance gene (macB) and fluoroquinolone antibiotic resistance genes (patA-B) in M. pneumoniae genome. In RMPP patients, higher relative abundance of Streptococcus pneumoniae had a strong correlation with increased hospitalization days while higher relative abundance of Streptococcus pneumoniae had a negative correlation with hospitalization days among NRMPP patients. CONCLUSIONS: The microbiota of LRT in children with RMPP was much more homogeneous and simpler than that of the NRMPP group and with lower relative abundance of macrolide antibiotics resistance gene in M. pneumoniae genome. M. pneumoniae was absolutely dominant in the vast majority of RMPP patients. Prolonged hospitalization days was associated with relative abundance of M. pneumoniae in NRMPP patients while it was related with other pathogens' relative abundance (e.g., Streptococcus pneumoniae) in RMPP patients.

The effect of vibrating positive expiratory pressure therapy on refractory Mycoplasma pneumoniae pneumonia prognosis in children.

BACKGROUND: It is well documented that vibration techniques and positive expiratory pressure (PEP) can improve airway clearance, however, few clinical trials have studied the efficacy of vibrating PEP therapy on refractory Mycoplasma pneumoniae (M. pneumoniae) pneumonia (RMPP) in children. This study aimed to explore using vibrating PEP therapy in children with RMPP. METHODS: Sixty participants in the remission stage of RMPP were recruited into this randomized study and divided into two groups. The Acapella group (n=30) used the Acapella® Choice twice daily for 2 months to clear their airways. The control group (n=30) used traditional chest percussion or postural drainage to mediate sputum expectoration. The groups administered their respective treatments at home and were followed up weekly. Participants kept a record of their treatment and condition in a daily log. The primary outcome of this study was the resolution of chest images, and the secondary outcome was the sputum period. RESULTS: There was no significant difference in sex or age distribution between the two groups. There were no significant differences at baseline between the groups in the location of their lesions (P=0.11). After 2 months of treatment, there was a better resolution of chest images in the Acapella group (P=0.00) compared with the control group. Additionally, the mean sputum period was significantly shorter for the Acapella group than for the control group (Acapella: 7.97±1.54 days; control: 11.90±1.64 days; P=0.00). CONCLUSIONS: Vibrating PEP therapy is an effective therapy for children with RMPP, both in airway clearance and the resolution of lung abnormalities.

Sleep-disordered breathing and genetic findings in children with Prader-Willi syndrome in China.

BACKGROUND: Sleep-related breathing disorders are common in individuals with Prader-Willi syndrome (PWS), and can include hypersomnolence and obstructive sleep apnea, as well as central sleep breathing abnormalities that are present from infancy. Here we describe the sleep-disordered breathing (SDB) and genetic findings in patients with PWS in China. METHODS: In all, 48 patients confirmed by genetic tests were enrolled, 32 were under 2 years of age and 16 were older children. There were 37 (77.1%) patients with paternal 15q11-13 deletions, 11 (22.9%) patients with maternal uniparental disomy (mUPD), and no patients with imprinting defect (ID). RESULTS: Compared with infants, a significantly higher proportion of older children with PWS were overweight or obese (15/16 children vs. 4/32 infants) and children had a higher serum level of free thyroxine (FT4) (0.9±0.2 vs. 0.7±0.7) and thyroxine (T4) (9.0±2.5 vs. 7.5±1.7). Age was correlated significantly with body mass index (BMI), T4, and FT4 (r=0.626, P=0.000; r=0.426, respectively). Overall, 42 of 48 (87.5%) patients had sleep apnea on polysomnography (PSG). Infants, when compared with older children, were more likely to experience central sleep apnea (71.8% vs. 25%). In infants, there were no significant differences in the prevalence of SDB between the deletion group and the mUPD group. CONCLUSIONS: Being overweight or obese was more common in older children with PWS. Compared with infants, a higher proportion children were overweight or obese and had higher serum levels of FT4 and T4. The prevalence of SDB was high in those with PWS, and central sleep apnea was found to be prevalent in infants. The pattern of SDB in infants with PWS was not significantly associated with the genotypes.

Exosomes secreted by endothelial progenitor cells improve the bioactivity of pulmonary microvascular endothelial cells exposed to hyperoxia in vitro.

BACKGROUND: Paracrine factors secreted by endothelial progenitor cells (EPCs) are suggested to be responsible, in part, for the improved microvascular development in bronchopulmonary dysplasia (BPD) models. This study aims to investigate the potential role of exosomes derived from EPCs (EPC-EXOs), a component of paracrine secretion, in angiogenesis by mediating the activity of PMVECs exposed to hyperoxia. METHODS: EPCs were isolated from bone marrow of rats. EPC-EXOs were isolated by ExoQuick-TC kits from the conditioned media of EPCs. The PMVECs were divided into three groups, including the normal group, the hyperoxia group (exposed to 85% O2) and the EPC-EXOs treatment group (exposed to 85% O2 and EPC-EXOs with the concentration of 100 µg/mL). The activities of proliferation, migration and tube formation of PMVECs were detected at the endpoint. The mRNA and protein expression levels of VEGF, VEGFR2 and eNOS in different groups were detected by real-time quantitative PCR and western blot. RESULTS: We found EPC-EXOs exhibited a cup or biconcave morphology, with the size ranging from 30 to 150 nm, and positive for the characteristic exosomal surface marker proteins, CD63 and TSG101. Comparing to the control group, Hyperoxic stress impaired the proliferation, migration, and tubule formation of PMVECs, and decreased the expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor 2 (VEGFR-2) of PMVECs. Comparing to the hyperoxia group, EPC-EXOs treatment enhanced the bioactivity of PMVECs in vitro, and increased the expression of eNOS, VEGF and VEGFR2. CONCLUSIONS: Our data demonstrate EPCs secrete exosomes that have independent angiogenic activity in vitro. This may help explain in part the protective effects of EPCs on hyperoxic injury in the developing lung vasculature and may represent a promising therapeutic strategy for BPD.

Clinical features of postinfectious bronchiolitis obliterans in children undergoing long-term nebulization treatment.

BACKGROUND: Limited data are available in relation to the clinical features of PIBO undergoing prolonged nebulization treatment with budesonide, terbutaline and ipratropium bromide. This retrospective study aimed to outline the features of clinical, high-resolution computed tomography (HRCT) and pulmonary function test (PFT) of PIBO, undergoing maintenance therapy utilizing a triple nebulization treatment and to determine the factors associated with prognosis. METHODS: Children diagnosed with PIBO were followed up between April 2014 and March 2017. The clinical features after maintenance nebulization treatment for 12 months were thereafter summarized. RESULTS: Thirty patients, 21 boys and 9 girls, were enrolled in the study. The median age of patients was 17.4 months, with a range between 3.0 and 33 months. Persistent coughing and wheezing were detected whilst wheezing and crackles were the common manifestations presented. HRCT scans revealed patchy ground and glass opacity, while PFT showed fixed airway obstruction in all patients. Four patients were lost during follow-up. After treatment, the clinical symptoms were improved greatly in all patients (P < 0.01). The mean increase in the percentage of TPEF%TE and VPEF%VE were improved greatly (P < 0.01). Images of the HRCT scan indicated marked improvements in 18 patients (81.8%) in comparison with scans obtained pre-treatment. CONCLUSIONS: Our data suggest a potential role of long-term nebulization treatment of budesonide, terbutaline, ipratropium bromide on PIBO, due to its efficacy as indicated in the improved clinical symptoms, pulmonary functions and CT manifestations identified in the children. New prospective and controlled studies are required to confirm this proposition.

Diagnostic Value of Nasopharyngeal Aspirates in Children with Lower Respiratory Tract Infections.

BACKGROUND: The accuracy of nasopharyngeal aspirate (NPA) specimens in detecting lower respiratory pathogens remains controversial. The objective of this study was to evaluate the diagnostic accuracy of aspirates (NPAs) specimen in lower respiratory tract infections (LRTIs) in children. METHODS: The prospective study was designed to collect the data of paired NPAs and bronchoalveolar lavage fluids from children with acute LRTIs from January 2013 to December 2015. All specimens were subjected to pathogen detection: bacterial detection by culture, Mycoplasma pneumoniae (Mp) detection by polymerase chain reaction assay and virus (influenza A and B viruses, parainfluenza virus [PIV] Types 1 and 3, respiratory syncytial virus, and adenovirus) detection by immunofluorescence assay. The diagnostic accuracy analysis of NPAs was stratified by age ≤3 years (n = 194) and >3 years (n = 294). RESULTS: We collected paired specimens from 488 children. The positive rate of pathogen was 61.6%. For Streptococcus pneumoniae, NPA culture had the specificity of 89.9% and negative predictive value of 100% in age ≤3 years, the specificity of 97.2% and negative predictive value of 98.9% in age >3 years. For Mp, the positive predictive values of NPA was 77.4% in children ≤3 years, and 89.1% in children >3 years. For PIV III, NPA specimen had the specificity of 99.8% and negative predictive value of 96.5% in children ≤3 years. For adenovirus, NPA had the specificity of 97.8% and negative predictive value of 98.4% in age ≤3 years, the specificity of 98.9% and negative predictive value of 99.3% in age >3 years. CONCLUSIONS: NPAs are less invasive diagnostic respiratory specimens, a negative NPA result is helpful in "rule out" lower airway infection; however, a positive result does not reliably "rule in" the presence of pathogens.

Influenza virus infection exacerbates experimental autoimmune encephalomyelitis disease by promoting type I T cells infiltration into central nervous system.

Multiple sclerosis starts with increased migration of auto-reactive lymphocytes across the blood-brain barrier, resulting in persistent neurodegeneration. Clinical and epidemiological studies indicated upper respiratory viral infections are associated with clinical exacerbation of multiple sclerosis. However, so far there is no any direct evidence to support it. Using the experimental autoimmune encephalomyelitis mice as the model for multiple sclerosis, we demonstrated that mice experienced with influenza virus infection were unable to recover from experimental autoimmune encephalomyelitis with a long-term exacerbation. The exacerbated disease was due to more type I T cells, such as CD45highCD4+CD44high, CD45highCD4+CCR5+, CD45high IFNγ+CD4+, MOG35-55-specific IFNγ+CD4+ and influenza virus-specific IFNγ+CD4+ T cells, infiltrating central nervous system in mice with prior influenza virus infection. Influenza virus infection created a notable inflammatory environment in lung and mediastinal lymph node after influenza virus inoculation, suggesting the lung may constitute an inflammatory niche in which auto-aggressive T cells gain the capacity to enter CNS. Indeed, the early stage of EAE disease was accompanied by increased CCR5+CD4+, CXCR3+CD4+ T cell and MOG35-55 specific CD4+ T cells localized in the lung in influenza virus-infected mice. CCL5/CCR5 might mediate the infiltration of type I T cells into CNS during the disease development after influenza infection. Administration of CCR5 antagonist could significantly attenuate the exacerbated disease. Our study provided the evidence that the prior influenza virus infection may promote the type I T cells infiltration into the CNS, and subsequently cause a long-term exacerbation of experimental autoimmune encephalomyelitis.

Interleukin-4 polymorphism is associated with severity of respiratory syncytial virus infection.

AIM: Respiratory syncytial virus (RSV) infection is a major health problem in young children. This study aimed to investigate whether the interleukin (IL)-4 -590C/T and -33 C/T polymorphisms were associated with the susceptibility to RSV infection in Chinese Han children and with the severity of the infection. METHODS: The IL-4 -590C/T and -33 C/T SNPs were evaluated in 218 inpatients with RSV bronchiolitis and 303 healthy controls. The severity of RSV bronchiolitis was evaluated using a respiratory scoring system. RESULTS: The frequencies of IL-4 -590 CC, CT and TT in the 218 RSV bronchiolitis patients versus the 303 controls were 1.4% versus 1.7%, 21.1% versus 34% and 77.5% versus 64.4%, respectively. The frequencies of IL-4 -33 CC, CT and TT were 1.4%, 20.6% and 78%, respectively, in RSV bronchiolitis patients and 2.3%, 35.6% and 62%, respectively, in the controls. The frequencies of the IL-4 -590C/T and -33 C/T polymorphisms were significantly different between the two groups. There was a significant difference in the frequency of the T-T haplotype formed by -590C/T and -33C/T between the two groups. The respiratory score of the RSV bronchiolitis cases with -590TT was 3.02 ± 0.44, which was significantly higher than those with -590CT (2.80 ± 0.44). The score for patients with -33TT was 3.03 ± 0.43, which was significantly higher than the score of 2.78 ± 0.46 for patients with -33CT. CONCLUSIONS: In Chinese Han children, the IL-4 -590C/T and -33 C/T polymorphisms were associated with the susceptibility to RSV and the severity of the infection.

Lactate Dehydrogenase as a Biomarker for Prediction of Refractory Mycoplasma pneumoniae Pneumonia in Children.

BACKGROUND: Corticosteroids have been used for refractory Mycoplasma pneumoniae pneumonia and have beneficial effects. The aim of this study was to identify the biomarkers for predicting refractory M. pneumoniae pneumonia in a timely fashion to initiate steroid therapy. METHODS: This was a prospective cohort study of children with M. pneumoniae pneumonia admitted to the Children's Hospital of Fudan University from September 2012 to August 2013. Lactate dehydrogenase (LDH) and other laboratory tests, including complete blood counts, C-reactive protein, erythrocyte sedimentation rate (ESR), alanine aminotransferase, aspartate aminotransferase, α-hydroxybutyrate dehydrogenase (HBDH), creatine kinase, and creatine kinase MB, were performed on admission. Based on the definition of refractory M. pneumoniae pneumonia, subjects were divided into 2 groups: refractory M. pneumoniae pneumonia and usual M. pneumoniae pneumonia. The diagnostic values of laboratory findings were analyzed. RESULTS: In total, 653 subjects were enrolled, including 300 in the refractory pneumonia group and 353 in the usual pneumonia group. There was no significant difference in sex distribution between the 2 groups. The average age in the refractory M. pneumoniae pneumonia group was greater than that in the usual M. pneumoniae pneumonia group. Compared with the usual pneumonia group, the refractory pneumonia group showed significantly higher levels of C-reactive protein, serum LDH, serum HBDH, serum alanine aminotransferase, serum aspartate aminotransferase, and neutrophils and higher ESRs. Logistic regression showed that age, LDH, and ESR were the significant factors in predicting refractory M. pneumoniae pneumonia. In addition, LDH and HBDH were strongly correlated, and receiver operating characteristic curve analysis showed that the area under the curve of LDH was 0.718 with a cutoff of 379 IU/L, that of ESR was 0.683 with a cutoff of 32.5 IU/L, and that of HBDH was 0.691 with a cutoff of 259.5 IU/L. CONCLUSIONS: Serum LDH can be used as a biomarker to predict refractory M. pneumoniae pneumonia at the early stage of hospitalization.

Combined iNO and endothelial progenitor cells improve lung alveolar and vascular structure in neonatal rats exposed to prolonged hyperoxia.

BACKGROUND: Stem cells or inhaled nitric oxide (iNO) are reported to improve lung structures in bronchopulmonary dysplasia (BPD) models. We hypothesized that combined iNO and transplanted endothelial progenitor cells (EPCs) might restore lung structure in rats after neonatal hyperoxia. METHODS: Litters were separated into eight groups: room air, hyperoxia, hyperoxia + iNO, hyperoxia + iNO + L-NAME, hyperoxia + EPCs, hyperoxia + EPCs + L-NAME, hyperoxia + EPCs + iNO, and hyperoxia + EPCs + iNO + L-NAME. Litters were exposed to hyperoxia from the 21st day, then, sacrificed. EPCs were injected on the 21st day. L-NAME was injected daily for 7 d from the 21st day. Serum vascular endothelial growth factor (VEGF), radial alveolar count (RAC), VIII factor, EPCs engraftment, lung VEGF, VEGFR2, endothelial nitric oxide (eNOS) and SDF-1 expression, and NO production were examined. RESULTS: Hyperoxia exposure led to air space enlargement, loss of lung capillaries, and low expression of VEGF and eNOS. Transplanted EPCs, when combined with iNO, had significantly increased engraftment in lungs, compared to EPCs alone, upon hyperoxia exposure. There was improvement in alveolarization, microvessel density, and upregulation of VEGF and eNOS proteins in the hyperoxia-exposed EPCs with iNO group, compared to hyperoxia alone. CONCLUSION: Combined EPCs and iNO improved lung structures after neonatal hyperoxia. This was associated with the upregulation of VEGF and eNOS expression.

The role of eNOS in the migration and proliferation of bone-marrow derived endothelial progenitor cells and in vitro angiogenesis.

The role of endothelial nitric oxide synthase (eNOS) in the activities of endothelial progenitor cells (EPCs) including migration, proliferation, and tube formation in vitro was investigated. EPCs were obtained from rat bone mononuclear cells by culturing for 7-10 days in EGM-2MV and identified by their capacity for FITC-UEA-1 binding and acetylated low-density lipoprotein (Dil-ac-LDL) intake using fluorescence microscopy. Migration, proliferation and tube formation activities were assessed in the presence or absence of N(ω)-nitro-L-argininemethylester (L-NAME), an eNOS inhibitor. mRNA and protein expression of CXCR4, CXCR7, VEGFR2, and eNOS were detected by real-time PCR and western blotting in the presence or absence of L-NAME. Nitric oxide production was detected by nitrate reductase in the presence or absence of L-NAME. Typical spindle-shaped cells appeared on the 7(th)-10(th) day and confluence reached about 80%. The percentage of FITC-UEA-1 and Dil-ac-LDL double-stained cells was about 85%. Cell migration, proliferation, and tube formation were significantly weakened after eNOS was inhibited (P < 0.05), and the expressions of CXCR4 and eNOS were significantly reduced (P < 0.05, respectively), but there was little change in CXCR7 and VEGFR2. NO production was dramatically decreased after eNOS was inhibited (P < 0.05). In summary, L-NAME significantly reduced the expression of eNOS and NO production by EPCs and inhibited migration, proliferation and tube formation by these cells, suggesting that eNOS affects EPC activities; CXCR4 may be implicated in the action of eNOS.

IFN-γ CA microsatellite polymorphism is associated with susceptibility to respiratory syncytial virus Infection and severity.

AIM: IFN-γ, an essential cytokine in the viral cell-mediated immune response, has been associated with the pathogenesis of respiratory syncytial virus (RSV) bronchiolitis and to the severity of the infection. The aim of this study was to investigate whether IFN-γ CA microsatellite (rs3138557) polymorphism was associated with susceptibility to RSV in Chinese Han children and with the severity of the infection. METHODS: The IFN-γ CA microsatellite was tested in 218 RSV bronchiolitis inpatients and 303 healthy controls, and the severity of the RSV bronchiolitis was evaluated using a standardised respiratory scoring system. RESULTS: The frequencies of CA12+/CA12+, CA12+/CA12- and CA12-/CA12- in the 218 RSV bronchiolitis patients and 303 controls were approximately 11% versus 19%, 55% versus 53% and 34% versus 28%, respectively. The gene polymorphism of IFN-γ CA repeats between the two groups was statistically different. The clinical respiratory scores of RSV bronchiolitis cases with CA12+/CA12+ and CA12+/CA12- were 2.84 ± 0.40 (SD) and 2.95 ± 0.44 (SD), respectively, and these were significantly lower than the 3.1 ± 0.36 (SD) score for those with CA12-/CA12-. CONCLUSION: IFN-γ CA microsatellite polymorphism was associated with the susceptibility of Chinese Han children to RSV and the severity of the infection.

Combined treatment for child refractory Mycoplasma pneumoniae pneumonia with ciprofloxacin and glucocorticoid.

OBJECTS: To evaluate the efficacy of combined treatment of ciprofloxacin and glucocorticoid for child refractory Mycoplasma pneumoniae (M. pneumoniae) pneumonia. METHODS: Clinical and laboratory characteristics of six pediatric refractory M. pneumoniae pneumonia cases treated with ciprofloxacin and glucocorticoids were reported. RESULTS: Five cases complicated with pleural effusion. The average febrile period prior to admission was 8.5 ± 2.0 days, the average total febrile period was 14.6 ± 7.6 days, and the average febrile period after treatment was 3.3 ± 3.0 days. The average time in hospital for patients was 23.6 ± 4.8 days. The initial mean WBC count was 10,100 ± 2,400/mm(3) . The erythrocyte sedimentation rate (ESR), lactate dehydrogenase (LDH), and C-reactive protein (CRP) in all cases were elevated during the course of illness. After combined therapy, all children clinically improved, with resolution of radiographic findings and normal laboratory items. We analyzed the 23S rRNA gene in four nasopharyngeal secretions, and found mutations in A2063G of domain V in three cases, and mutation in A2064G in the other case. CONCLUSION: Combined treatment of ciprofloxacin and glucocorticoids can significantly ameliorate child refractory M. pneumoniae pneumonia further comparative study is needed to well evaluate the treatment efficacy.

Clinical study of 28 cases of paediatric idiopathic pulmonary haemosiderosis.

OBJECTIVE: To summarize the clinical characteristics of idiopathic pulmonary haemosiderosis (IPH) to explore the aetiopathogenesis, risk factors, diagnosis and experiences in therapy of IPH. METHODS: The documents of 28 IPH cases, who were hospitalized in Children's Hospital of Fudan University between February 1989 and June 2009 were reviewed. RESULTS: (i) fifteen cases were males and 13 were females, and 88.5% of the cases had first onset under the age of 10 years; (ii) the triad occurred in 57.1% cases; (iii) radiographic features of IPH including diffuse alveolar-type infiltrates, ground glass attenuation, interstitial reticular and micronodular patterns; (iv) haemosiderin-laden macrophages were found in 60.7% of the cases;(v) the trend of positive correlation was found between the severity of ventilatory restrictive pattern and the disease courses (r = 0.229, p = 0.237); and (vi) glucocorticosteroids can control the symptoms. CONCLUSION: (i) the clinical presentations are not classical. If long-term anaemia exists without reason, this case must be considered; (ii) corticosteroid can control the symptom; and (iii) IPH may be associated with the imbalance of immune system.

Haplotype of IL-8 -251T and 781C is associated with the susceptibility to respiratory syncytial virus.

OBJECTIVE: To study the association of haplotypes of IL-8 -251T/A and 781 C/T single nucleotide polymorphisms (SNPs) with the susceptibility of respiratory syncytial virus (RSV). METHODS: This study included 101 hospitalized patients under 2 years who suffered from RSV pneumonia and 108 hospitalized patients under 2 years who suffered only from pneumonia without RSV infection. Genotypes of two SNP loci in all enrolled persons were defined by allele-specific polymerase chain reaction. The allele's frequencies of SNPs were analyzed with case-control study, linkage of two loci and haplotypes composed by the two loci were also studied. RESULTS: (i) The frequency of IL-8 -251T in cases was dramatically high (OR = 2.08, p = 0.0002). (ii) Haplotype of TC was significantly high in cases (p = 0.01). CONCLUSION: These findings support that haplotype of TC composed by IL-8 -251T and 781C is associated with the susceptibility of RSV.