OBJECTIVETo investigate the effect of respiratory syncytial virus (RSV)-related pulmonary infection on endogenous metabolites in large intestinal mucosa in BALB/c mice using metabolomics technology based on gas chromatography-mass spectrometry (GC-MS).

METHODSMice were randomly divided into a control group and a RSV pneumonia model group (n=16 each). The mouse model of RSV pneumonia was established using intranasal RSV infection (100×TCID, 50 μL/mouse, once a day). After 7 days of intranasal RSV infection, the mice were sacrificed and GC-MS was used to identify endogenous metabolites and measure the changes in their relative content in colon tissue. SMCA-P12.0 software was used to perform principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) for endogenous metabolites in colon tissue. The differentially expressed metabolites in colon tissue were imported into the metabolic pathway platform Metaboanalyst to analyze related metabolic pathways.

RESULTSPCA and OPLS-DA showed significant differences between the control and RSV pneumonia model groups. A total of 32 metabolites were identified in the colon tissue of the mice with RSV pneumonia. The RSV pneumonia model group had significant increases in the content of leucine, isoleucine, glycine, alanine, arachidonic acid, and lactic acid, which were related to the valine, leucine, isoleucine, arachidonic acid, and pyruvic acid metabolic pathways.

CONCLUSIONSRSV pneumonia might cause metabolic disorders in the large intestinal tissue in mice.

Amino Acids, Branched-Chain ; metabolism ; Animals ; Female ; Gas Chromatography-Mass Spectrometry ; Intestinal Mucosa ; metabolism ; Intestine, Large ; metabolism ; pathology ; Lung ; pathology ; Mice ; Mice, Inbred BALB C ; Pneumonia, Viral ; metabolism ; Respiratory Syncytial Virus Infections ; metabolism

No abstract available.
Air Pollutants/*toxicity ; Animals ; Biological Markers/metabolism ; Cell Proliferation/*drug effects ; Female ; Immunohistochemistry ; Inhalation Exposure ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa/drug effects/pathology ; Ozone/*toxicity ; Proliferating Cell Nuclear Antigen/metabolism ; Respiratory Mucosa/*drug effects/metabolism/pathology ; Skin/*drug effects/metabolism/pathology

OBJECTIVE: To study the expression of epidermal growth factor receptor (EGFR), C-erbB-2 and its relationship with cell proliferation in nasopharyngeal carcinoma. METHOD: Expression of C-erbB-2, EGFR and proliferating cell nuclear antigen (PCNA) were detected with immunohistochemical staining in 32 nasopharyngeal carcinoma samples and 12 chronic inflammatory nasopharyngeal tissue samples. RESULT: The positive rate of EGFR,C-erbB-2, and PCNA expression in nasopharyngeal carcinoma was 65.6%, 37.5%, and (42.5 +/- 22.6)%, respectively, which was significantly higher than that in chronic inflammatory nasopharyngeal tissue (P < 0.05). There were positive correlations between the positive rate of EGFR, C-erbB-2, and PCNA expression and histopathological stage. The co-expression of C-erbB2 and EGFR was found in 62.5% (20/32) nasopharyngeal carcinoma samples. There was a positive correlation between C-erbB-2 and EGFR expression (r = 0.38, P < 0.05). The highest percentage of PCNA expression was found in carcinoma samples with co-expression of C-erbB and EGFR. CONCLUSION C-erbB-2, EGFR might have synergetic effect in the development and progress of nasopharyngeal carcinoma. The co-expression of C-erbB-2 and EGFR closely correlates with cell proliferation status.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Proliferation ; ErbB Receptors ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Receptor, ErbB-2 ; metabolism ; Respiratory Mucosa ; metabolism ; Young Adult

OBJECTIVETo study the alternative expression and sequence of human elongation factor-1 delta (human EF-1 delta p31) during malignant transformation of human bronchial epithelial cells induced by cadmium chloride (CdC12) and its possible mechanism.

METHODSTotal RNA was isolated at different stages of transformed human bronchial epithelial cells (16HBE) induced by CdCl2 at a concentration of 5.0 microM. Special primers and probe for human EF-1 delta p31 were designed and expression of human EF-1 delta mRNA from different cell lines was detected with fluorescent quantitative PCR technique. EF-18 cDNA from different cell lines was purified and cloned into pMD 18-T vector followed by confirming and sequencing analysis.

RESULTSThe expressions of human EF-1 beta p31 at different stages of 16HBE cells transformed by CdCl2 was elevated (P < 0.01 or P < 0.05). Compared with their corresponding non-transformed cells, the overexpression level of EF-1 delta p31 was averagely increased 2.9 folds in Cd-pretransformed cells, 4.3 folds in Cd-transformed cells and 7.2 folds in Cd-tumorigenic cells. No change was found n the sequence of overexpressed EF-1beta p31 at different stages of 16HBE cells transformed by CdCl2.

CONCLUSIONOverexpression of human EF-1beta p31 is positively correlated with malignant transformation of 16HBE cells induced by CdC12, but is not correlated with DNA mutations.

Cadmium Chloride ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Peptide Elongation Factor 1 ; genetics ; metabolism ; Respiratory Mucosa ; drug effects ; metabolism ; pathology ; Sequence Analysis, DNA

The EGFR plays an essential role in goblet cell hyperplasia and mucus hypersecretion. EGFR has an intrinsic tyrosine kinase activity that, when activated, induces the production of MUC5AC through the signaling kinase cascade in the airway epithelium. We have investigated the effects of an EGFR tyrosine kinase inhibitor, gefitinib, on ovalbumin (OVA)-induced, allergic inflammation in airway epithelia of mice. OVA-sensitized mice were pretreated with gefitinib at two different doses (12.5 and 50 mg/kg) and then challenged with OVA. The OVA challenge increased the total cell count and eosinophil count in bronchoalveolar lavage fluid (BALF), as well as the concentrations of T-helper2 (Th2) cytokines, such as IL-4 and IL-13, overall eosinophil recruitment in the lung tissue and airway hyperresponsiveness (AHR). Pretreatment with gefitinib reduced the inflammatory cell counts and released cytokine concentrations (IL-4 and IL-13) in BALF, as well as eosinophil recruitment in the lungs and AHR, in a dose-dependent manner. This was associated with decreased EGFR and Akt phosphorylation. We showed that gefitnib inhibits EGFR and phosphoinositol 3'-kinase (PI3K)/Akt activation which were activated in OVA sensitized mice. These findings suggest that inhibitors of the EGFR cascade may have a role in the treatment of asthma.
Animals ; Antineoplastic Agents/*therapeutic use ; Bronchoalveolar Lavage Fluid/cytology ; Cytokines/biosynthesis ; Enzyme Activation ; Eosinophils/cytology ; Goblet Cells/pathology ; Inflammation/drug therapy/metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; Phosphorylation ; Proto-Oncogene Proteins c-akt/metabolism ; Quinazolines/*therapeutic use ; Receptor, Epidermal Growth Factor/*antagonists & inhibitors/metabolism ; Respiratory Hypersensitivity/*drug therapy/etiology/metabolism ; Respiratory Mucosa/drug effects/pathology

Airway structural changes that occur in patients with asthma in response to persistent inflammation are termed airway remodeling. The cysteinyl leukotrienes (LTC4, D4 and E4) are known to play important roles in the pathobiology of asthma. To evaluate the effect of low dose montelukast (MK) on the development of airway remodeling using a chronic murine model of allergic airway inflammation with subepithelial fibrosis, BALB/c mice, after intraperitoneal ovalbumin (OVA) sensitization on days 0 and 14, received intranasal OVA periodically on days 14-75. MK treated mice received montelukast sodium intraperitoneally on days 26-75. The OVA sensitized/challenged mice developed an extensive eosinophil cell inflammatory response, goblet cell hyperplasia, mucus occlusion, and smooth muscle hypertrophy of the airways. In addition, in OVA sensitized/challenged mice, dense collagen deposition/fibrosis was seen throughout the lung interstitium surrounding the airways, blood vessels, and alveolar septae. The cysteinyl leukotriene 1 (CysLT1) receptor antagonist, MK significantly reduced the airway eosinophil infiltration, goblet cell hyperplasia, mucus occlusion, and lung fibrosis except airway smooth muscle hypertrophy in the OVA sensitized/challenged mice. The OVA sensitized/challenged mice had significantly increased epithelial desquamation compared with control mice. MK markedly reduced epithelial desquamation of airways in OVA/MK treated animals compared with OVA sensitized/challenged mice. MK treatment did not affect the levels of CysLT in lung tissue. Our results show that the important role of cysteinyl leukotrienes in the pathogenesis of asthma. Lower dose of CysLT1 receptor antagonism has a significant anti-inflammatory effect on allergen-induced lung inflammation and fibrosis but not airway smooth muscle hypertrophy in an animal model of asthma.
Respiratory Mucosa/pathology ; Receptors, Leukotriene/metabolism ; Quinolines/*therapeutic use ; Pulmonary Fibrosis/pathology ; Muscle, Smooth/pathology ; Mucus/secretion ; Mice, Inbred BALB C ; Mice ; Lung/pathology ; Leukotrienes/*biosynthesis ; Leukotriene Antagonists/*therapeutic use ; Hypertrophy ; Hyperplasia ; Goblet Cells/pathology ; Drug Evaluation, Preclinical ; Dose-Response Relationship, Drug ; Disease Models, Animal ; Cysteine/*biosynthesis ; Collagen/metabolism ; Asthma/*drug therapy/metabolism/pathology ; Anti-Asthmatic Agents/*therapeutic use ; Animals ; Airway Obstruction/drug therapy/pathology ; Acetates/*therapeutic use

OBJECTIVEThis study was designed to establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from human lung squamous cell carcinoma tissue and paired tumor-adjacent normal bronchial epithelial tissue, and to identify differential expression of tumor-associated proteins by using proteome analysis.

METHODSComparative proteome analysis of human lung squamous carcinoma and paired normal bronchial mucosa adjacent to tumors from 20 cases were carried out. Total proteins of the carcinoma tissue and normal bronchial mucosa were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTS(1) Seventy-six differentially expressed proteins were screened by analyzing the electrophoretic maps of the 20 carcinoma and control mucosa tissues. (2) Sixty-eight differential proteins were identified by peptide mass fingerprinting (PMF). Some proteins were products of oncogenes and others were involved in the regulation of cell cycle and signal transduction. (3) The expression of three proteins mdm2, c-Jun and EGFR, correlated with lung squamous carcinoma, were detected by immunohistochemical staining and Western blot analysis. The results showed that the expression of mdm2, c-Jun and EGFR were up-regulated in lung squamous carcinomas, whereas down-regulated in control normal mucosa. It was consistent with our proteome analysis results. Those results suggested that those proteins may play roles in the carcinogenesis of lung squamous carcinoma.

CONCLUSIONsixty-eight differentially expressed proteins were successfully characterized by comparative proteome analysis. Those results may provide scientific foundation for screening the molecular biomarkers which can be used in diagnosis and treatment of lung squamous carcinoma, as well as to improve patients' prognosis and provide a new clue for carcinogenesis research of lung squamous cell carcinoma.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Proteome ; Proto-Oncogene Proteins c-jun ; metabolism ; Proto-Oncogene Proteins c-mdm2 ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Respiratory Mucosa ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Up-Regulation

OBJECTIVEIt is supposed that bronchial epithelial cells responses to the environmental stimuli are different between asthmatic and non-asthmatic individuals, which contribute to the pathogenesis of asthma. These different responses produce different mediators. If differential gene expressions are found in bronchial epithelial cells of asthmatic and non-asthmatic individuals after the same stimuli in vitro, and these genes are overexpressed in asthmatic children in vivo, then it is concluded that these genes may be associated with asthma. Therefore the authors analyzed the differential gene expressions in the bronchial epithelium cells of asthmatic and non-asthmatic children after RSV infection in vitro. Among these genes, Galectine-7 (lectin, galactoside-binding, soluble, 7, Galectin-7) was 8 times up-regulated in asthmatic children. Galectine-7 was associated with skin keratinocyte apoptosis. The authors hypothesized that Galectin-7 may also be associated with bronchial epithelial cell apoptosis in asthmatic children. The aim of this study was to understand the role of Galectine-7 in bronchial epithelial cell apoptosis in asthma.

METHODSThe bronchial mucosae of one asthmatic child and one non-asthmatic child were obtained by biopsy and cultured in vitro. The bronchial epithelial cells were infected by RSV. The differential gene expressions were analyzed with micro array. Among those differentially expressed genes, Galectin-7 was 8 times up-regulated in asthmatic children. The bronchial mucosae from 10 asthmatic children and 17 non-asthma children were investigated for cell DNA break, Galectine-7 and mRNA expression, Caspase-3 expression by TUNEL, hybridization in situ and immunochemistry. Image analysis was used for quantitative assessment.

RESULTSGalectine-7 gene was 8 times up-regulated in bronchial epithelial cells from asthmatic children after RSV infection in vitro. Galectin-7 and mRNA were overexpressed in bronchial epithelial cells in asthma in vivo. Bronchial epithelial cell apoptosis increased in asthma in vivo.

CONCLUSIONGalectin-7 may be associated with bronchial epithelial cell apoptosis in asthma.

Adolescent ; Apoptosis ; genetics ; Asthma ; metabolism ; pathology ; Biopsy ; Bronchi ; metabolism ; pathology ; Bronchoscopy ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Child ; Child, Preschool ; Epithelial Cells ; metabolism ; pathology ; virology ; Female ; Galectins ; genetics ; metabolism ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; In Situ Hybridization ; In Situ Nick-End Labeling ; Infant ; Male ; RNA, Messenger ; Respiratory Mucosa ; cytology ; metabolism ; pathology ; virology ; Respiratory Syncytial Viruses ; pathogenicity ; Up-Regulation

OBJECTIVETo investigate the relationship of BCL-10 protein and API2-MALT1 fusion gene in MALT lymphoma.

METHODSSpecimens from 86 cases of MALT lymphoma were studied by immunohistochemical staining for BCL-10. RT-PCR was used to detect the transcripts of API2-MALT1 fusion gene.

RESULTSIn all 10 cases of Hashimoto thyroiditis only cytoplasmic BCL-10 expression in lymphoid cells was observed. In 86 MALT lymphoma cases, 42 cases (48. 8%) exhibited BCL-10 expression in both nucleus and cytoplasm. API2-MALT1 fusion gene was detected in 35 cases (40. 7%) of MALT lymphoma. BCL-10 nuclear expression was correlated with API2-MALT1 fusion gene transcript (r = 0. 374,P = 0. 000).

CONCLUSIONBCL-10 nuclear expression is correlated with API2-MALT1 fusion gene expression in MALT lymphoma.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; B-Cell CLL-Lymphoma 10 Protein ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Female ; Follow-Up Studies ; Gastric Mucosa ; metabolism ; pathology ; Hashimoto Disease ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Lymphoid Tissue ; metabolism ; pathology ; Lymphoma, B-Cell, Marginal Zone ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Respiratory Mucosa ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the role of matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor (TIMP-1) in the pathogenesis of bronchial asthma and assess the effect of steroid treatment on MMP-9 and TIMP-1 levels. Matrix metalloproteinases are a family of zinc and calcium-dependent endopeptidases. Many MMPs such as MMP-1, MMP-2, MMP-3 are associated with asthma, in which MMP-9 is the key factor in asthma. Tissue inhibitor-1 of metalloproteinases is a specific inhibitor of MMP-9; the MMP-9 and TIMP-1 imbalance could lead to airway inflammation and remodeling in lung disease such as asthma.

METHODSForty Wistar rats were divided into 4 groups randomly: control, asthma model 7 days (7-day group), asthma model 21 days (21-day group) and steroid treatment groups. Asthma model of rats were established by ovalbumin (OVA) sensitization and challenge with mist inhalation. The expression of MMP-9 and TIMP-1 in lung tissues was detected by immunocytochemistry, RT-PCR and Western blotting.

RESULTS(1) By observing the changes of action, tracing respiratory curves, detecting level of serum IgE level and observing the lung tissues sections, the authors demonstrated that the rat asthmatic models were successfully established. (2) The lung tissue sections of the asthma groups stained with hematoxiline and eosin (HE) showed many inflammatory cell infiltrations around the bronchioli and accompanying arterioles, hyperplasia of caliciform cells, broken bronchial mucous membrane and thickening of submucosal layer. The hyperplasia of airway smooth muscle and basement membrane were more significant in asthma model 21-day group than that in 7-day group. These changes were improved after treatment. (3) The expression of MMP-9 in rat's lung tissues: the expression was 2.71 +/- 0.37 in 7-day group, 1.76 +/- 0.27 in 21-day group, 0.88 +/- 0.18 in the treatment group and 0.52 +/- 0.10 in the control group (F = 151.52, P < 0.01). The expression of TIMP-1 in rat's lung tissues was 1.13 +/- 0.19 in the 7-day group, 1.55 +/- 0.24 in 21-day group, 0.77 +/- 0.15 in the treatment group and 0.47 +/- 0.08 in the control group (F = 69.46, P < 0.01). (4) The results of immunocytochemistry and protein expression were consistent with those of RT-PCR.

CONCLUSIONThe protein and mRNA expression level of MMP-9 and TIMP-1 was high in asthmatic rat's lung tissues. Down-regulation of the expression of MMP-9 and TIMP-1 by steroids may be one of the mechanisms by which airway inflammation and remodeling are inhibited in asthma.

Administration, Inhalation ; Animals ; Asthma ; drug therapy ; metabolism ; Blotting, Western ; Bronchi ; metabolism ; pathology ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Enzymologic ; Glucocorticoids ; administration & dosage ; therapeutic use ; Immunohistochemistry ; Inflammation ; drug therapy ; metabolism ; pathology ; Lung ; drug effects ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Rats ; Rats, Wistar ; Respiratory Mucosa ; drug effects ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism