Inhibition of M2 macrophage polarization and reduction of airway inflammation in asthmatic mice with lncRNA MRAK088388 knockout / 细胞与分子免疫学杂志

Objective To investigate the long non-coding RNA(lncRNA) MRAK08838 regulates macrophage function to influence the development of asthmatic airway inflammation. Methods MRAK088388 gene knockout (MRAK088388-/-) mouse model was prepared and allergic asthma was induced by dust mite protein Dermatophagoides farinae 1 (Der f1). The mice were sacrificed after 28 days of modeling, and serum was collected to measure IgE and IgG. The FinePointe RC system was used to measure airway hyperresponsiveness and evaluate lung function in mice. Lung tissue was taken for HE staining, and periodic acid-Schiff (PAS) staining was used to evaluate inflammatory infiltration and mucus secretion in mouse lungs. Fluorescence quantitative PCR was used to detect the expression level of lncRNA MRAK08838 in bronchoalveolar lavage fluid (BALF) cells and lung tissue of asthmatic mice. ELISA was used to detect the levels of inflammatory cytokines IFN-γ, IL-4, IL-5, IL-13, IL-10 and IL-17A. Flow cytometry was used to evaluate the phenotype of macrophages in BALF and lung tissue, as well as the proportion of neutrophils, eosinophils, and alveolar macrophages. The changes of the above indicators were detected in mice by adoptive transfer of bone marrow-derived macrophages (BMDM). Results Under the challengle of Der f1, MRAK088388-/- mice showed reduced allergic airway inflammation, including reduced eosinophils in BALF and reduced production of IgE and IgG1. In addition, Der f1-treated MRAK088388-/- mice had fewer M2 macrophages than wild-type asthmatic mice. Wild-type mouse BMDM (M0) and Der f1-treated MRAK088388-/- mice also showed mild inflammatory response. Conclusion Knockout of MRAK088388 alleviates airway inflammation in asthmatic mice by inhibiting M2 polarization of airway macrophages.

Long noncoding RNA MRAK088388 is involved in the progression of childhood allergic asthma via miR-30a/ATG5 axis / 西安交通大学学报(医学版)

【Objective】 To investigate the association between the long-stranded non-coding ribonucleic acid (lncRNA) MRAK088388 and allergic asthma in children. 【Methods】 A total of 15 healthy children and 15 children with asthma were monitored for disease progression over a 2-year period. Blood samples were collected from patients during the chronic phase of the disease for lncRNA/mRNA expression microarray analysis. Competing endogenous RNA networks (MRAK088388/miR-30a/ATG5) were identified by bioinformatics analysis. In vitro cultured bronchial epithelial (16HBE) cells were used to quantify gene and associated protein expression levels by real-time fluorescent quantitative polymerase chain reaction (qPCR) and protein blotting, respectively. Cell Counting Kit-8 and transwell assays were used to assess the proliferation and migration of 16HBE cells and verify the effects of MRAK088388, miR-30a and ATG5 on asthma. 【Results】 Six lncRNA-miRNA-mRNAs were identified by correlation analysis. By qRT-PCR analysis, MRAK088388/miR-30a/ATG5 was selected to construct the ceRNA network in this study. mRAK088388 and ATG5 expressions were high in the peripheral blood of children with asthma, while the expression of miR-30a was low (P<0.05). The expression level of E-cadherin was significantly higher in 16HBE cells after si-MRAK088388+TGF-β1 group, while the expression levels of Vimentin and α-SMA were significantly lower (P<0.05), indicating that knockdown of MRAK088388 inhibited the epithelial mesenchymal transition in 16HBE cells. Compared with si-NC+ TGF-β1 group, the cell morphology of si-MRAK088388+TGF-β1 group was similar to that of the control group, indicating that MRAK088388 knockdown attenuated TGF-β1-induced cell morphological changes; in addition, MRAK088388 knockdown inhibited TGF-β1-induced proliferation and migration of 16HBE cells. MRAK088388 was confirmed by qPCR and protein blotting to promote the progression of childhood asthma by targeting the miR-30a/ATG5 axis. 【Conclusion】 Childhood asthma is associated with the MRAK088388/miR-30a/ATG5 axis, and MRAK088388 is involved in the process of childhood allergic asthma by negatively regulating miR-30a expression and regulating elevated ATG5 expression levels to affect bronchial epithelial cell mesenchymal transition, proliferation, and migration.

Effects of vitamin C on A549 cell proliferation, apoptosis and expressions of Caspase, Survivin / 中国肺癌杂志

<p><b>BACKGROUND AND OBJECTIVE</b>It was proven that Vitamin C could inhibit the growth of many types of tumors as an antioxidant. The aim of this study is to explore role of Vitamin C in proliferation and apoptosis of lung carcinoma cell line A549 and the underlying mechanism.</p><p><b>METHODS</b>A549 cells were cultured in vitro and incubated with Vitamin C. The cell viability was measured by growth curve and clonogentic assay. Flow cytometry was used to analyze cell cycle and detect apoptosis. The levels of expression of Caspase-3 mRNA and Survivin mRNA were detected by RT-PCR.</p><p><b>RESULTS</b>Vitamin C of 400 microg/mL, 4 mg/mL significantly inhibited the growth of A549 cell lines (P = 0.024, P = 0.015, respectively). Flow cytometry showed that the cells major stagnation stayed in the G0/G1 and S phase and the apoptotic rate increased with time prolonged. Vitamin C signifiantly up-regulated the expression of Caspase-3 mRNA, but had no effect on Survivin mRNA.</p><p><b>CONCLUSION</b>Vitamin C can inhibit the proliferation of A549, block A549 cells in G0/G1 and S phase, and induce apoptosis of A549 cells. Apotosis occurred by up-regulated the expression of Caspase-3.</p>

Molecular epidemiology of respiratory syncytial virus in children with acute respiratory tract infections and sequence analysis of G gene in Chongqing area / 中华微生物学和免疫学杂志

Objective To investigate the epidemiological features of respiratory syncytial virus (RSV) A and B in hospitalized children with acute respiratory tract infections(ARTIs) and to analyze the genetic characteristics of G protein gene of RSV in Chongqing area, especially for BA strains. Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTls from April, 2008 to March, 2009 were screened for RSV using RT-PCR. Full length G protein gene was amplified by RT-PCR from 10 RSV subtype A and 29 RSV subtype B strains. Results Out of the total 508 specimens, 126 (24.8%) were revealed positive for RSV. RSV subtype A and B viruses accounted for 34.1% and 63.5% of the total positive specimens, respectively. The remaining 2.4% of the specimens were positive for both subtype A and B. At the nucleotide level, identities between the 10 subtype A virus G genes and that of the prototype strain A2 were 91.4% -92.0%, indicating genotype GA2. Identities of the 29 subtype B virus G genes and that of the CH18537 strain were 92.0%-93.0%. Nineteen out of 29 RSV subtype B isolates contained highly repeated 60 nucleotides insertion in G protein gene, namely BA strain. As compared to the prototypes, the RSV G protein gene included nucleotide deletion, insertion, substitutions, especially in the carboxy-terminal third of the G gene. Condnsion RSV has been the major cause of acute respiratory tract infections in children in Chongqing area. Subtype B strains, especially BA strains, were predominant during the study peried. Whether the predominated circulation of BA strain is resulted from enhanced attachment function of G protein remains unknown.

Analysis of pulmonary alveolar proteinosis in recent 20 years / 中国综合临床

Objective To analyze the epidemiology,clinical and imageological characteristics,diagnosis,misdiagnosis,treatment and prognosis of pulmonary alveolar proteinosis (PAP) reported in China in recent 20 years,in an attempt to provide important clues for prompt and accurate diagnosis of PAP.Methods Clinical data of PAP from 1988 to 2008 in China were retrospectively analyzed and the clinical data of 126 patients with PAP which were misdiagnozed were summarized.Results There were 19 cases misdiagnozed with pneumonia,24 cases with pulmonary cancer,18 cases with bronchitis,19 cases with idiopathic pulmonary fibrosis,15 cases with pulmonary tuberculosis,10 cases with eosinophilic pneumonia,9 cases with sarcoidosis,5 cases with fungus pneumonia.The clinical manifestations of PAP had no specificity and the imageology manifestation was various.Its final diagnosis mainly depended on the examination of brochoalveolar lavage fluid and/or lung biopsy,and pathologic examination.Conclusions The diversity of clinical manifestations of PAP has resulted in higher clinical misdiagnosis rate.WhoLe lung irrigation is the safest and the most effective way to treat PAP.