[Association of Haemophilus influenzae infection with environmental and climatic factors in Suzhou, China]. / è‹å·žåœ°åŒºæµæ„Ÿå—œè¡€æ†èŒæ„ŸæŸ“ä¸ŽçŽ¯å¢ƒæ°”å€™çš„ç›¸å ³å› ç´ åˆ†æž.

OBJECTIVES: To investigate the epidemiological characteristics of respiratory Haemophilus influenzae (HI) infection in children in Suzhou, China and its association with climatic factors and air pollutants. METHODS: The data on air pollutants and climatic factors in Suzhou from January 2016 to December 2019 were collected. Respiratory secretions were collected from 7 940 children with acute respiratory infection who were hospitalized during this period, and bacterial culture results were analyzed for the detection of HI. A stepwise regression analysis was used to investigate the association of HI detection rate with air pollutants (PM2.5, PM10, NO2, SO2, CO, and O3) and climatic factors (monthly mean temperature, monthly mean humidity, monthly total rainfall, monthly total sunshine duration, and monthly mean wind speed). RESULTS: In 2016-2019, the 4-year overall detection rate of HI was 9.26% (735/7 940) among the children in Suzhou. The children aged <1 year and 1-<3 years had a significantly higher HI detection rate than those aged ≥3 years (P<0.01). The detection rate of HI in spring was significantly higher than that in the other three seasons, and the detection rate of HI in autumn was significantly lower than that in the other three seasons (P<0.001). The multiple linear regression analysis showed that PM10 and monthly mean wind speed were independent risk factors for the detection rate of HI: the detection rate of HI was increased by 0.86% for every 10 µg/m3 increase in the concentration of PM10 and was increased by 5.64% for every 1 m/s increase in monthly mean wind speed. Air pollutants and climatic factors had a lag effect on the detection rate of HI. CONCLUSIONS: HI is an important pathogen for acute respiratory infection in children in Suzhou and is prevalent in spring. PM10 and monthly mean wind speed are independent risk factors for the detection rate of HI.

Solitary primary pulmonary synovial sarcoma: A case report.

BACKGROUND: Synovial sarcoma (SS) is an uncommon and highly malignant soft tissue sarcoma in the clinic, with primary pulmonary SS (PPSS) being extremely rare. Here, we describe the clinical characteristics, diagnosis, and treatment of a solitary PPSS case confirmed via surgical resection and fluorescence in situ hybridization (FISH). CASE SUMMARY: A 33-year-old man was admitted because of intermittent coughing and hemoptysis for one month, with lung shadows observed for two years. Whole-body positron emission tomography-computed tomography (PET-CT) revealed a solitary mass in the upper lobe of the right lung, with uneven radioactivity uptake and a maximum standardized uptake value of 5.6. The greyish-yellow specimen obtained following thoracoscopic resection was covered with small multi-nodulated structures and consisted of soft tissue. Hematoxylin and eosin staining revealed spindle-shaped malignant tumor cells. Immunohistochemistry indicated these tumor cells were CD99 and BCL-2-positive. Furthermore, the FISH test revealed synovial sarcoma translocation genetic reassortment, which confirmed the diagnosis of SS. CONCLUSION: PPSS is extremely rare and tends to be misdiagnosed as many primary pulmonary diseases. PET-CT, histologic analysis, and FISH tests can be used to differentiate PPSS from other diseases. Surgical resection is regularly recommended for the treatment of solitary PPSS and is helpful for improving the prognosis.

The expression and function of programmed death-ligand 1 and related cytokines in neutrophilic asthma.

BACKGROUND: Programmed death-ligand 1 (PD-L1) is an important immune checkpoint inhibitor. Recent studies suggest that the PD-L1-mediated pathway may be a promising target in allergic asthma. However, the mechanism by which PD-L1 represses neutrophilic asthma (NA) remains unclear. In this study, we examined correlations between the expression of PD-L1 and the production of T helper cell type 1 (Th1), T helper cell type 2 (Th2), and T helper cell type 17 (Th17) cells in pediatric patients with NA and a mouse model. METHODS: The clinical samples of 26 children with asthma and 15 children with a bronchial foreign body were collected over a period of 12 months by the Children's Hospital of Soochow University. An experimental mouse model of asthma was established to study NA. An enzyme-linked immunoassay (ELISA) was used to assess soluble PD-L1 (sPD-L1) and cytokines [e.g., interleukin (IL)-4, IL-6, interferon gamma (IFN-γ), IL-17 and granulocyte-macrophage colony-stimulating factor (GM-CSF)] in bronchoalveolar lavage fluid (BALF). RESULTS: NA patients had significantly higher levels of sPD-L1, IL-6, IL-17, and GM-CSF in their BALF than non-NA and control patients (P<0.05). In a murine model of asthma, the positive rate and fluorescence intensity of PD-L1 in the NA group and the immunoglobulin G (IgG)-treated NA group were higher than in the PD-L1 antibody (Ab)-treated NA group and the phosphate-buffered saline (PBS) control group (P<0.05). In the plasma and the BALF of the NA group and the IgG-treatment NA group, the levels of IL-17, IL-4, tumor necrosis factor alpha (TNF-α), and granulocyte colony-stimulating were higher than those in the PBS control group (P<0.05). The histopathological examination of lung tissues from all mice groups showed that a large number of inflammatory cells were found around the airway in the NA group and the IgG-treatment group. CONCLUSIONS: PD-L1 may contribute to the Th17/IL-17 immune response, which is associated with neutrophilic inflammation and asthma. A PD-L1 blockade reduces pulmonary neutrophils and mucus production.

The diagnostic value of serological tests and real-time polymerase chain reaction in children with acute Mycoplasma pneumoniae infection.

BACKGROUND: This study set out to evaluate the clinical significance and diagnostic effectiveness of serological tests and real-time polymerase chain reactions (RT-PCR) in children of different age groups and disease durations infected with Mycoplasma pneumoniae (MP). METHODS: Pediatric patients with lower respiratory tract infection (LRTI) confirmed by polymerase chain reaction (PCR) were enrolled and subjected to bronchoalveolar lavage fluid PCR (BALF-PCR) for MP infection. The diagnostic values of the serum immunoglobulin M (IgM) test, paired sera immunoglobulin G (IgG) test, RT PCR applied to nasopharyngeal aspirates (NPA-PCR), and combined IgM and NPA-PCR test were evaluated. RESULTS: When BALF PCR was used as the gold standard, the MP positivity rate of combined IgM and NPA PCR was 78.85%in children aged 3-5 years. The positivity rates of IgM, NPA PCR, and combined IgM and NPA PCR in children older than 5 years were 71.21%, 72.72%, and 84.85%, respectively. The detection rate of combined IgM and NPA PCR was consistent with BALF PCR (Kappa =0.727). The MP positivity rates of combined IgM and NPA PCR at 1-2 weeks was as high as 91.11%, and was consistent with the BALF PCR (Kappa =0.756). Moreover, the positivity rates of IgM or NPA PCR at 2-3 weeks were 63.16%, and were consistent with each other (Kappa =0.771). CONCLUSIONS: Combined IgM and NPA PCR is the optimal test to confirm MP infection among children aged 3-5 years in cases with a disease duration of less than2 weeks, and either NPA PCR or IgM is recommended for children older than 5 years with a disease duration of 2-3 weeks. KEYWORDS: Mycoplasma pneumoniae pneumonia (MPP); diagnosis; children; age; disease duration.

Molecular characterization of a RNA polymerase (RNAP) II (DNA directed) polypeptide H (POLR2H) in Pacific white shrimp (Litopenaeus vannamei) and its role in response to high-pH stress.

RNA polymerase (RNAP) II (DNA-directed) (POLR2) genes are essential for cell viability under environmental stress and for the transfer of biological information from DNA to RNA. However, the function and characteristics of POLR2 genes in crustaceans are still unknown. In the present study, a POLR2H cDNA was isolated from Pacific white shrimp (Litopenaeus vannamei) and designated as Lv-POLR2H. The full-length Lv-POLR2H cDNA is 772 bp in length and contains a 32-bp 5'- untranslated region (UTR), a 284-bp 3'- UTR with a poly (A) sequence, and an open reading frame (ORF) of 456 bp encoding an Lv-POLR2H protein of 151 amino acids with a deduced molecular weight of 17.21 kDa. The Lv-POLR2H protein only contains one functional domain, harbors no transmembrane domains and mainly locates in the nucleus. The expression of the Lv-POLR2H mRNA was ubiquitously detected in all selected tissues, with the highest level in the gills. In situ hybridization (ISH) analysis showed that Lv-POLR2H was mainly located in the secondary gill filaments, the transcript levels of Lv-POLR2H in the gills were found to be significantly affected after challenge by pH, low salinity and high concentrations of NO2- and NH4+, indicating that Lv-POLR2H in gill tissues might play roles under various physical stresses. Specifically, under high-pH stress, knockdown of Lv-POLR2H via siRNA significantly decreased the survival rate of the shrimp, indicating its key roles in the response to high-pH stress. Our study may provide the first evidence of the role of POLR2H in shrimp responding to high-pH stress and provides new insight into molecular regulation in response to high pH in crustaceans.

[Detection of viral pathogens and allergens in infants and young children at high risk of asthma during a wheezing episode].

OBJECTIVE: To investigate the viral etiology and allergen distribution in infants and young children at high risk of asthma during a wheezing episode. METHODS: A total of 135 infants and young children at high risk of asthma were enrolled who were admitted due to asthmatic bronchitis or asthmatic bronchopneumonia between April 2016 and August 2017. Fluorescent probe PCR was used to measure influenza A (Flu A), respiratory syncytium virus (RSV), adenovirus (ADV), parainfluenza virus (PinF), human rhinovirus (HRV), human partial lung virus (hMPV) and human bocavirus (HBoV) in nasopharyngeal aspirates. ImmunoCAP was used to measure inhaled allergens, food allergens, and total IgE concentration. RESULTS: Among the 135 patients, the overall virus detection rate of nasopharyngeal aspirates was 49.6%, and HRV had the highest detection rate of 25.2%, followed by HBoV (9.6%), RSV (8.1%), PinF (5.9%), Flu-A (3.7%), ADV (1.5%) and hMPV (0.7%). The 1-3 years group had a significantly higher detection rate of HRV than the <1 year group (P<0.05). The positive rate of allergen screening was 59.3%, with 44% for inhaled allergens and 89% for food allergens. Among the inhaled allergens, dust mites had the highest positive rate of 77%, followed by mould (37%), pollen (26%) and animal dander (9%). Among the food allergens, egg white had a positive rate of 73% and milk had a positive rate of 68%. The <1 year group had a significantly higher positive rate of inhaled allergens than the 1-3 years group (P<0.05). The 1-3 years age group had a significantly higher level of T-IgE than the <1 year group (P<0.05). The positive virus group had a significantly higher positive rate of inhaled allergens than the non-virus group (P<0.05). The children with the second wheezing episode had significantly higher positive rates of inhaled allergens and food allergens and level of T-IgE than those with the first wheezing episode (P<0.05). The children with the second wheezing episode also had significantly higher positive rates of dust mites and mould than those with the first wheezing episode (P<0.05). CONCLUSIONS: Early HRV infection and inhaled allergen sensitization are closely associated with the development of wheezing in infants and young children at high risk of asthma.

The role of miR-29c/B7-H3/Th17 axis in children with Mycoplasma pneumoniae pneumonia.

BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) is one of the most common causes of community-acquired pneumonia in children. Recent studies demonstrated that the incidence of severe or fatal M. pneumoniae was gradually increasing, which may be related to the excessive inflammation. However, the exact pathogenesis of excessive inflammation in Mycoplasma pneumoniae pneumonia(MPP) is still unclear. This study aimed to reveal the role of miR-29c/B7-H3/Th17 axis in children with MPP. METHODS: Children hospitalized in Respiratory Department during Jan. 2014 to Dec. 2015 were enrolled. All children enrolled was confirmed with MP infection using real-time PCR and ELISA. Children were excluded if they were co-infected with other pathogens. A total of 52 children with MPP and 26 controls were enrolled. miR-29c expression in monocytes of children with MPP was determined by real-time PCR and soluble B7-H3 (sB7-H3) and IL-17 were determined by ELISA, and explore their clinical significance. miR-29c overexpression and silencing technology and luciferase reporter assay were performed to confirm whether B7-H3 is the direct target of miR-29c. The levels of transcription factor ROR-γt in CD4+ T cells and cytokine IL-17A in supernatant were detected after stimulated by different concentrations of B7-H3 fusion protein in vitro. RESULTS: Of all 52 children with MPP, the mean age of the children were 77 ± 33 months, and 23 cases were male accounting for 44.2%. Nineteen cases had pleural effusion accounting for 36.5%. Children with MPP had significantly lower level of miR-29c and higher level of sB7-H3 and IL-17 compared to controls (both P < 0.05). The level of miR-29c significantly increased during convalescent phase compared to that of acute phase while sB7-H3 and IL-17 significantly decreased during convalescent phase (both P < 0.05). There was a positive correlation between the level of sB7-H3 and IL-17 in children with MPP during acute-stage (r = 0.361,P = 0.009). Children with MPP combined with pleural effusion had significantly higher level of sB7-H3 compared to those without pleural effusion (9952.3 ± 3065.3 vs. 7449.7 ± 2231.5, pg/ml), and the levels of sB7-H3 was positively correlated with the number of days of fever. The level of miR-29c was negatively correlated with M. pneumoniae specific IgG, IgM level. High concentrations of B7-H3(15µg/ml) could enhance ROR-γt expression and increase IL-17A. Functional studies based on luciferase reporter assay and immunofluorescence staining suggested that B7-H3 is the direct target of miR-29c, and miR-29c silencing or overexpression could up- or down-regulate the expression of B7-H3 in THP-1 cells. CONCLUSIONS: The axis of miR-29c/B7-H3/Th17 plays a vital role in children with MPP through excessive inflammation. miR-29c and B7-H3 may be the new target for the prevention and treatment of MPP, and may be the novel and potential biomarkers for the assessment of prognosis.

Rapid development of metastatic pulmonary calcifications in primary hyperparathyroidism: a case report and literature review.

BACKGROUND: Metastatic pulmonary calcification (MPC) is rarely reported in primary hyperparathyroidism, especially MPC develops quickly. We report such a case here with a literature review. CASE PRESENTATION: A 41-year-old woman presented with cough and dyspnea. Data from clinical, radiological, pathological, technetium (99mTc)-methylene diphosphonate (MDP) bone scintillation imaging, and 99mTc-methoxy isobutyl isonitrile (MIBI) thyroid imaging were studied. 99mTc-MIBI thyroid imaging indicated hyperparathyroidism. Chest computed tomography (CT) scans showed rapidly progressive bilateral pulmonary multiple high-density shadows with mass consolidation and exudation in only five days. 99mTc-MDP bone scintillation imaging indicated bilateral pulmonary calcifications. CT-guided lung biopsy showed multifocal irregularities of calcium deposition and calcified bodies in the pulmonary interstitium. The patient showed gradually clinical and radiological improvement after surgical removal of the parathyroid adenoma. CONCLUSION: Rapidly progressive MPC tends to be misdiagnosed as many primary pulmonary diseases. 99mTc-MDP bone scintillation imaging and pulmonary biopsy could be performed to differentiate metastatic pulmonary calcification from other diseases. Surgical resection of the parathyroid gland is helpful for treatment of MPC in patients with primary hyperparathyroidism and is regularly recommended.

The pulmonary nodule "discovered" by pneumonia: a case report.

The number of patients diagnosed with pulmonary nodules increased as more patients with high risk of lung cancer choose low-dose computed tomography (CT) scans for the screening of cancer. Clinicians might get two questions from the patients: what is the definite diagnosis of the nodule? What should we do? We have already got many guidelines trying to solve these problems. There are also several prediction models for pulmonary nodules. However, guidelines are not suitable for all types of patients, and the reality of patients is more complicated. Here we reported a 58-year-old man with a lung nodule in the right upper lobe, which was occasionally found during a period of pneumonia. We suggested two periods of follow-up, and the patient was also admitted to a clinical trial about circulating tumor cells (CTCs). He finally accepted surgical excision with a pathologic diagnosis of adenocarcinoma. This case suggests that: we might suggest CT surveillance for patients with solid nodules about 8 mm maximum diameter; three-dimensional reconstruction of CT scans could provide more information about the details of nodules; CTCs counts of peripheral blood could be considered as a potential clue for malignancy.

[Association of T lymphocyte subsets and allergens with Mycoplasma pneumoniae infection complicated by wheezing in infants and young children].

OBJECTIVE: To investigate the percentage of T lymphocyte subsets and allergen screening results in infants and young children with Mycoplasma pneumoniae (MP) infection complicated by wheezing. METHODS: Flow cytometry was used to measure the percentage of peripheral blood T cell subsets in 354 infants and young children with MP infection complicated by wheezing (MP wheezing group), 336 infants and young children with MP infection but without wheezing (MP non-wheezing group), and 277 children with recurrent wheezing (recurrent wheezing group). Allergen screening was also performed for these children. RESULTS: Both the MP wheezing group and recurrent wheezing group had significantly lower percentages of CD3+ and CD3+CD8+ lymphocytes than the MP non-wheezing group (p<0.05). The MP groups with or without wheezing had a significantly higher percentage of CD3+CD4+ lymphocytes than the recurrent wheezing group (p<0.05). Both the MP wheezing group and recurrent wheezing group had significantly higher percentages of CD3-CD19+ and CD19+CD23+ lymphocytes than the MP non-wheezing group (p<0.05), and the recurrent wheezing group had the highest percentages (p<0.05). The overall positive rate of food allergens was significantly higher than that of inhaled allergens (30.3% vs 14.7%; p<0.05). The positive rates of food and inhaled allergens in the recurrent wheezing group and MP wheezing group were significantly higher than in the MP non-wheezing group (p<0.05), and the recurrent wheezing group had the highest rates. CONCLUSIONS: Imbalance of T lymphocyte subsets and allergic constitution play important roles in the pathogenesis of MP infection complicated by wheezing in infants and young children.

Acute fibrinous and organizing pneumonia: A case report and literature review.

Acute fibrinous and organizing pneumonia (AFOP) is a rare lung disease with distinct histological characteristics that include the diffuse presence of intra-alveolar fibrin, and the absence of eosinophils and hyaline membrane. In the present study, a case of AFOP that was diagnosed by lung biopsy is described. The patient presented with high fever and a cough with expectoration. Computed tomography of the lung showed the presence of bilateral patchy infiltrates, predominantly in the lower lobes. Histopathological examination of lung biopsy from the lower pulmonary lobe confirmed the pathological diagnosis. The patient showed a poor response to treatment with prednisone. Based on a review of literature pertaining to documented AFOP cases, a summary of the clinical features, radiological characteristics, treatment outcomes and prognoses associated with AFOP are presented. The most common pulmonary symptoms included cough, dyspnea and fever. The primary imaging findings in AFOP were consolidation and ground-glass opacity in the bilateral lung.

[Etiology of acute respiratory tract infection in hospitalized children in Suzhou from 2005 to 2011].

OBJECTIVE: To elucidate the etiology of acute respiratory tract infection (ARI) in hospitalized children in Suzhou from 2005 to 2011. METHODS: A total of 10 243 hospitalized children with ARI in Children's Hospital Affiliated to Soochow University from September 2005 to October 2011 were enrolled in the study. The clinical information was collected; and the nasopharyngeal aspiration fluid and serum samples were sent for multi-pathogen detection. Respiratory syncytial virus (RSV), influenza virus type A and B (IV-A, IV-B), parainfluenza virus type 1-3 (PIV-1-PIV-3) and adenovirus (ADV) were detected by direct immunofluorescence assay. Human bocavirus (HBoV), mycoplasma pneumoniae (MP) and chlamydia pneumoniae (CP) were detected by fluorescent quantitative PCR while human metapneumovirus (hMPV) was detected by reverse transcription PCR (RT-PCR). Sputum culture was applied to detect bacterial infection and quantitative ELISA was adopted to detect the specific antibodies of MP and CP. The results of the above detections were analyzed, and thereby to explore the prevalent pathogens among different aging children and the seasonal distribution and characteristics of the disease. RESULTS: At least one type of pathogen was detected in 5871 out of 10 243 hospitalized children and the overall positive rate was 57.32%; including 3326 virus samples with positive rate at 32.47% (3326/10 243), 2870 bacteria samples with positive rate at 28.02% (2870/10 243) and 2759 atypical pathogen samples,with positive rate at 26.94% (2759/10 243). MP was the most common pathogen,whose detected rate was 25.74% (2637/10 243). The median age of children with RSV (6 months) or PIV-3(8 months) infection was younger than the median age of all hospitalized children (12 months) (χ(2) = 380.992, 34.826, P < 0.05). While the median age of children with ADV (42 months), HBoV (14 months) or IV-A (24 months) infection was older than it of all hospitalized children (χ(2) = 83.583, 13.169, 18.012, P < 0.05). The median age of children with MP (30 months),streptococcus pneumoniae (17 months) or haemophilus parainfluenzae (21 months) infection was older than it of all hospitalized children (χ(2) = 728.299, 60.463, 8.803, P < 0.05). The detected rate of RSV in the groups of children aging less than 6 months, 7-12 months, 2-3 years, 4-5 years and over 6 years was separately 25.59% (840/3283), 17.05% (333/1953), 11.85% (310/2615), 6.68% (90/1347), and 2.87% (30/1045); which decreased while the age grew (χ(2) = 178.46, P < 0.01). Conversely, the positive rate of MP increased with the age growing (χ(2) = 379.21, P < 0.01). The rate in the above groups was 8.25% (271/3283), 19.46% (380/1953), 33.00% (863/2615), 41.43% (558/1347), 54.07% (565/1045), respectively. RSV and IV-A were prevalent in winter, whose detected rates were 35.73% (941/2634) and 4.44% (117/2634) respectively.hMPV infection was common in spring, with the detected rate at 10.55% (278/2634); while HBoV infection was common in summer and autumn, with the positive rate at 9.99% (149/1491) and 9.71% (98/1009). MP and CP were frequently detected in summer, up to 31.27% (819/2619) and 10.07% (43/427) respectively. RSV was the most common pathogen in bronchiolitis (33.27% (866/2603)) and MP was the most common pathogen in bronchopneumonia (26.05% (1152/4422)) and lober pneumonia (52.25% (267/511)). CONCLUSION: MP and RSV were the most common pathogens in respiratory tract infection in hospitalized children. The novel virus included hMPV and HBoV, which also played an important role in ARI. Different pathogens were prevalent in different ages; with respective seasonal distribution and characteristics.

[Invasive aspergillosis after near-drowning: case reports and review of the literature].

OBJECTIVE: The purpose of this study was to analyze the clinical presentations and disease courses of invasive aspergillosis (IA) in patients after near-drowning. METHODS: The clinical data of 3 cases of invasive aspergillosis after near-drowning from Oct. 2005 to Aug. 2010 in this hospital were retrospectively analyzed, and the related literature was reviewed. RESULTS: There were 1 male and 2 female patients, aged from 18 to 72 years. All of them had been immunocompetent before drowning. Two patients drowned because of traffic accident, and 1 fell in sewage by accident. All of the 3 patients were intubated because of acute respiratory failure, and received broad-spectrum antibiotic therapy. One had transient leucopenia, and 2 patients received glucocorticoid therapy. The condition of the 3 cases deteriorated 9 to 11 days after near-drowning. Aspergillus was isolated from sputum samples of 2 patients at the same time. Thoracic CT findings included multiple nodules, consolidation and cavity formation. Multiple abscesses in cerebral parenchyma were found in 1 patient with invasive cerebral aspergillosis. One patient died, whose lungs, cerebral parenchyma, myocardium and kidney were all infected by aspergillus. The other 2 patients, whose infection limited to the lungs, had a positive prognosis. Using the terms "aspergillosis" and "near-drowning" a PUBMED search yielded 7 articles, published between 1984 and 2010. Using the terms "invasive pulmonary aspergillosis" and "near-drowning", searching Wangfang data and CHED data, encompass 1 article, published in 2009. In all of the 8 articles, there are 5 final diagnosis cases and 3 clinical diagnosis cases. CONCLUSIONS: IA was very rare in immunocompetent hosts but had been reported in previously healthy individuals after near-drowning. Aspergillosis might develop 1 to 2 weeks after near-drowning, and the prognosis was poor in patients with central nervous system involvement.

Inhibition of pre-B cell colony-enhancing factor attenuates inflammation and apoptosis induced by pandemic H1N1 2009 in lung endothelium.

The recent pandemic influenza A (H1N1 2009) virus infection has caused acute lung injury in susceptible population resulting in high mortality in ICU patients. In this report, we observed the effect of pre-B cell colony-enhancing factor (PBEF) on the inflammation and apoptosis in H1N1-infected human pulmonary microvascular endothelial cells (HPMECs). We constructed an in vitro HPMEC monolayer model. The results showed that H1N1 2009 induced the increased expression of inflammatory cytokines (IL-6/IL-8/TNF-α/IP-10) and apoptosis factors (FasL/TRAIL) in infected HPMECs. However, PBEF silencing with siRNA inhibited the expression of some inflammatory cytokines and decreased the apoptosis mediated by FasL. We conclude that PBEF might be partially responsible for the localized inflammatory response to H1N1 2009 in the lung microvascular endothelium and the H1N1-induced endothelial cell apoptosis probably through the FasL-mediated pathway.

[Effect of heparin upon inflammatory reaction of endotoxin-induced acute lung injury in rat].

OBJECTIVE: To investigate the effects of heparin upon inflammatory reaction and associated mechanism of endotoxin-induced acute lung injury (ALI) in rat. METHODS: Thirty-six male Sprague-Dawley rats were randomly divided into three equal groups namely: ALI group, heparin treatment group and normal control group. The ALI rats were induced by injecting endotoxin intravenously and sacrificed at 4 h after model establishment. The lung histology was scored by a modification of Smith technique. The albumin permeability of pulmonary microvascular (P(alb)) was measured by single nuclide tracer technique. Tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6) and von Willebrand factor (vWF) levels of serum were determined using commercial enzyme-linked immunosorbent assay kits. The expressions of lung tissue extacellular signal-regulated kinases (ERK)-1/2, P38 mitogen-activated protein kinase (MAPK) and c-jun N-terminal kinases (JNK) were determined by Western blotting. RESULTS: The Smith lung injury score in heparin treatment group and ALI group were (5.00 +/- 1.26) and (8.00 +/- 1.09) respectively. The values were significantly higher than that of normal control group (0.67 +/- 0.52, both P < 0.01). However, the Smith lung injury score in heparin treatment group was significantly lower than that of ALI group (P < 0.01). The P(alb), TNF-alpha, IL-6 and vWF of heparin treatment group were (0.28 +/- 0.04), (1.92 +/- 0.35) microg/L, (1.22 +/- 0.13) ng/ml and (24.9 +/- 4.0) U/L respectively. The values were significantly higher than those of normal control group [0.20 +/- 0.02, (0.51 +/- 0.09) microg/L, (0.23 +/- 0.05) ng/ml and (14.0 +/- 3.0) U/L respectively, all P < 0.01] but significantly lower than those of ALI group [(0.38 +/- 0.04), (2.77 +/- 0.37) microg/L, (1.62 +/- 0.13) ng/ml and (31.8 +/- 7.5) U/L respectively, all P < 0.01]. The lung tissue levels of phospho-ERK1/2 and phospho-P38 MAPK expressions of heparin treatment group were markedly higher than those of normal control group, whereas markedly lower than those of ALI group. There was no marked difference of phospho-JNK expression in all three groups. CONCLUSION: Heparin markedly inhibits the expressions of phospho-ERK1/2 and phospho-P38 MAPK, down-regulates the inflammatory reaction, attenuates the endothelial permeability of pulmonary vasculature and significantly improves endotoxin-induced lung injury in rats.

Antithrombin-III without concomitant heparin improves endotoxin-induced acute lung injury rats by inhibiting the activation of mitogen-activated protein kinase.

BACKGROUND: Antithrombin-III (AT-III), the major inhibitor of thrombin in plasma, also has anti-inflammation property and might have positive effect on sepsis. The present study aimed to investigate the effects of AT-III on inflammatory reaction and pulmonary protection in endotoxin-induced acute lung injury (ALI) rat. METHODS: Sixty male Sprague-Dawley rats were randomly assigned equally to normal control group, ALI group, AT-III treatment group, AT-III + heparin treatment group, and heparin treatment group. The pulmonary vascular permeability index (PVPI) was measured by single nuclide tracer technique. The activity of AT-III in plasma was determined by the method of synthetic chromogenic substrata. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in serum were determined by enzyme-linked immunosorbent assay. The expressions of lung tissue mitogen-activated protein kinases (ERK1/2, P38 and JNK MAPK) were determined by Western blotting. RESULTS: Rats had significantly improved lung histopathology in the AT-III treatment group and heparin treatment group compared with the ALI group. The PVPI of the ALI group was 0.38 + or - 0.04, significantly higher than that of the normal control group (0.20 + or - 0.02, P < 0.01), AT-III treatment group (0.30 + or - 0.04, P < 0.01) and heparin treatment group (0.28 + or - 0.04, P < 0.01) respectively. There were no significant differences of PVPI in the ALI group and AT-III + heparin treatment group. The activity of AT-III in plasma in the ALI group was (76 + or - 8)%, significantly lower than that of the normal control group ((96 + or - 11)%, P < 0.05) and AT-III treatment group ((105 + or - 17)%, P < 0.05) respectively. The serum levels of TNF-alpha and IL-6 of the ALI group were (2.770 + or - 0.373) microg/L and (1.615 + or - 0.128) ng/ml respectively, significantly higher than those of the normal control group ((0.506 + or - 0.093) microg/L and (0.233 + or - 0.047) ng/ml respectively, all P < 0.01), AT-III treatment group ((1.774 + or - 0.218) microg/L and (1.140 + or - 0145) ng/ml respectively, all P < 0.01) and heparin treatment group ((1.924 + or - 0.349) microg/L and (1.223 + or - 0.127) ng/ml respectively, all P < 0.01). The lung tissue levels of phospho-ERK1/2 and phospho-P38 MAPK expressions were markedly higher in the ALI group than in the normal control group, AT-III treatment group and heparin treatment group respectively. CONCLUSIONS: AT-III without concomitant heparin inhibited the activation of ERK1/2 and P38 MAPK, down-regulated the levels of downstream cytokines TNF-alpha and IL-6, relieved endothelial permeability, and improved the ALI in endotoxin-induced rats. It might be helpful to administrate AT-III alone, not with concomitant heparin, to those patients with ALI and sepsis.

[Effect of beta-adrenergic agonist on alveolar fluid clearance in acute lung injury: an experiment with rats].

OBJECTIVE: To determine the effect of dobutamine, a beta-adrenergic agonist, on the alveolar fluid clearance (AFC) in acute lung injury (ALI). METHODS: Thirty two male Sprague-Dawley rats were randomly divided into four equal groups: normal control group; ALI group, infused with endotoxin to induce ALI; dobutamine control group, receiving sustained intravenous injection of dobutamine at the dose of 5 microg/kg/min, and dobutamine treatment group, receiving sustained intravenous injection of dobutamine at the dose of 5 microg/kg/min after the administration of endotoxin. Experiment began 45-60 minutes after the circulation was stable. Blood pressure was measured and blood gas analysis was conducted at the beginning of the experiment and one hour later. Perfusion fluid with (125)I-albumin with the radioactivity of 1.5 microCi/ml was perfused into the lung. One hour after the mechanical ventilation the mice were killed and their lungs were taken out. Alveolar fluid was taken out to calculate the AFC by single nuclide tracer technique. RT-PCR was used to detect the expression of alpha, beta, and gamma-rat epithelial sodium channel (rENaC) mRNA. RESULTS: (1) The AFC of ALI group was 14.0 +/- 1.2%, significantly lower than that of the normal control group (21.0 +/- 3.9%, P < 0.05). (2) The AFC of the dobutamine control group was 26.6 +/- 1.6%, significantly higher than that of the normal control group (P < 0.05). and the AFC of the dobutamine treatment group was 20.0 +/- 3.9%, significantly higher than that of the ALI group (P < 0.05). (3) The expression of a-rENaC mRNA was 1.40 +/- 0.40 in the ALI group and was 1.38 +/- 0.13 in the dobutamine treatment group, both significantly higher than those in the dobutamine control group (1.01 +/- 0.14) and in the normal control group (0.44 +/- 0.11, all P < 0.05). The expression of beta-rENaC mRNA was 0.70 +/- 0.8 in the ALI group was 0.71 +/- 0.17 in the dobutamine treatment group, both significantly higher than those in the dobutamine control group (0.58 +/- 0.12) and in the normal control group (0.44 +/- 0.11, all P < 0.05). There were not significant differences in expression of alpha and beta-rENaC mRNA between the normal control group and dobutamine control group, and between the ALI group and dobutamine treatment group (both P > 0.05). (4) The gamma-rENaC mRNA expression was 0.90 +/- 0.19 in the dobutamine control group and was 0.97 +/- 0.15 in the dobutamine treatment group, both significantly higher than that in the normal control group (0.69 +/- 0.10) and in the ALI group (0.70 +/- 0.32) (all P < 0.05). CONCLUSION: Able to upregulate the gamma-rENaC expression and improve the AFC in acute lung injury, beta-adrenergic agonist may be beneficial to reduce lung edema in ALI patients.

[Change of extravascular lung water in sheep with early acute respiratory distress syndrome].

OBJECTIVE: To determine the change of extravascular lung water (EVLW) and the effect of positive end-expiratory pressure (PEEP) and cardiac output (CO) on EVLW in the sheep with early acute respiratory distress syndrome (ARDS). METHODS: The sheep ARDS model was induced by infusion of endotoxin intravenously, and then divided into ARDS model group (n = 12), PEEP group (n = 21) and dobutamine group (n = 13). The PEEP group was further divided into three subgroups: PEEP 5 cm H(2)O (n = 7), 10 cm H(2)O (n = 8) and 15 cm H(2)O groups (n = 6). EVLW was measured by the single indicator thermodilution technique. The EVLW, hemodynamics and lung mechanics parameters were observed at 6 h after sheep ARDS model was induced, 2 h after mechanical ventilation with PEEP (5, 10, 15 cm H(2)O) respectively and 2 h after CO increase >/= 50% of the base value (induced by dobutamine infusion). RESULTS: (1) When ARDS was induced in sheep, EVLW increased from (12.8 +/- 4.7) ml/kg to (18.1 +/- 7.1) ml/kg (P < 0.01), and EVLW had no significant change (P > 0.05) during the early phase of ARDS (6 h). PaO(2)/FiO(2) was (136.8 +/- 34.9) mm Hg at ARDS 0 h, which was higher than that before infusion of endotoxin [(444.3 +/- 127.7) mm Hg, P < 0.01]. EVLW had no relationship with PaO(2)/FiO(2), but DeltaEVLW after ARDS model induced had good correlation with DeltaPaO(2)/FiO(2) (r = -0.501, P < 0.001). When ARDS was induced, static lung compliance decreased from 26.9 +/- 8.4 to 16.4 +/- 5.0 (P < 0.01). (2) After mechanical ventilation with PEEP 2 h, EVLW in PEEP 10 cm H(2)O group and 15 cm H(2)O group were (14.7 +/- 4.5) ml/kg and (15.3 +/- 3.7) ml/kg respectively, which was significantly less than that before PEEP [(16.5 +/- 4.7) ml/kg and (18.4 +/- 6.0) ml/kg respectively, all P < 0.05]. But EVLW in 5 cm H(2)O group showed no difference during 2 h PEEP application. (3) After CO increase >/= 50% of the base value 1 h and 2 h, EVLW was (16.3 +/- 4.9) ml/kg and (16.9 +/- 6.9) ml/kg respectively, which did not differ from that of the baseline value [(15.1 +/- 4.6) ml/kg, all P > 0.05]. CONCLUSIONS: EVLW in ARDS sheep increased significantly and remained at the same level during the early phase of ARDS. PEEP had a markedly effect in reducing EVLW, while the increase of CO induced by dobutamine did not significantly raise EVLW.

[Effects of positive end-expiratory pressure on lung recruited volume and oxygenation in patients with acute respiratory distress syndrome].

OBJECTIVE: To access alveolar recruitment with positive end-expiratory pressure (PEEP) and to correlate the recruited volume with arterial oxygenation in acute respiratory distress syndrome (ARDS). METHODS: Eleven ventilated patients satisfying criteria of ARDS were included in the study group. Recruited volume of three different PEEP levels [5 cm H2O, 10 cm H2O, 15 cm H2O (1 cm H2O=0.098 kPa)] were measured by pressure-volume curve method. Effects of different PEEP levels on lung mechanics and gas exchange were compared. RESULTS: Lung recruited volume elevated when PEEP increased from 5 cm H2O to 15 cm H2O[(40.2+/-15.3)ml vs. (123.8+/-43.1)ml vs. (178.9+/-43.5)ml, all P<0.05]. Arterial oxygenation index increased with PEEP and a positive correlation was found between recruited volume and changes in arterial oxygenation index (r=0.483, P<0.01). There was no significant difference during the static compliance at different PEEP levels at a fixed tidal volume (P>0.05). Recruited volume at PEEP 15 cm H2O in the patient group with lower inflection point (LIP) was larger than the group without LIP. CONCLUSION: Recruited volume increases with PEEP, and the PEEP-induced increase in arterial oxygenation is significantly correlated to recruited volume.

[A comparison study on quantitative methods for the recruited volume in sheep with acute respiratory distress syndrome].

OBJECTIVE: To compare equal pressure method with pressure-volume curve method to quantify the recruited volume. METHODS: Acute respiratory distress syndrome sheep model was induced by intravenous infusion 3 microg/kg lipopolysaccharide (LPS). Recruited volume of three different levels of positive end-expiratory pressure (PEEP=5, 10, 15 cm H2O) were measured both by pressure-volume curve method and by equal pressure method. RESULTS: The time needed to measure recruited volume by pressure-volume curve method was 5-6 minutes, which was longer than that of equal pressure method. Recruited volume measured by the two methods increased with PEEP. No significant difference was found between the recruited volume measured by equal pressure method and by equal pressure method at PEEP of 5 cm H2O, they were (25.79+/-20.48) ml vs. (63.26+/-54.57) ml (P>0.05), while recruited volume at PEEP of 10 cm H2O and 15 cm H2O measured by equal pressure method were lower than those measured by the pressure-volume curve method, they were (48.64+/-30.51)ml vs. (148.14+/-85.42)ml and (71.50+/-58.09)ml vs. (322.86+/-148.42)ml (all P<0.05) respectively. CONCLUSION: Though equal pressure method is simple, it could not take the place of pressure-volume curve method to quantify recruited volume.