PURPOSE:To investigate whether airway eosinophilic degranulation develops in Mycoplasma pneumonia (M. pneumonia), and to elucidate the association between M. pneumonia and asthma. METHODS:Forty patients with M. pneumonia, 20 stable asthma patients (stable asthma) and 20 normal controls were recruited from October 2005 to February 2007. In the M. pneumonia, blood and induced sputum sampling were collected at admission (acute stage) and 6 to 8 weeks later (convalescent stage). Eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) levels in sputum and serum were measured in all 3 groups. RESULTS:Serum levels of EDN and ECP in the acute stage of M. pneumonia were comparable to those in the stable asthma group. However, in the convalescent stage of M. pneumonia, EDN and ECP levels were significantly lower than in the stable asthma (P<0.01 and P<0.05, respectively). Sputum levels of EDN and ECP levels in the acute stage of M. pneumonia were comparable to those in the stable asthma. Sputum EDN levels in the convalescent stage of M. pneumonia were significantly lower than those in the stable asthma (P<0.05), and sputum ECP levels were lower than those in the stable asthma, which was not statistically significant. CONCLUSION:Eosinophilic degranulation may play an important role in the pathogenesis of M. pneumonia, which suggests the association between M. pneumonia and asthma.
Asthma ; Eosinophil Cationic Protein ; Eosinophil-Derived Neurotoxin ; Eosinophils ; Humans ; Inflammation ; Mycoplasma ; Pneumonia ; Pneumonia, Mycoplasma ; Sputum

PURPOSE: Eosinophilic inflammation plays a critical role in asthma and high-resolution computed tomography (HRCT) scoring systems have been used to evaluate the extent and severity in long standing adult asthma. We investigated if there is a correlation between eosinophil degranulation markers and HRCT scores in childhood asthma. METHODS: Children with acute asthma exacerbation (n=25) underwent HRCT and were assessed for bronchial wall thickening (BWT), low lung density (LLD), and bronchial dilatation (BD) using semi-quantitative scoring techniques. Serum eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) levels indicating eosinophil degranulation were determined. Comparisons were made with normal control subjects (n=14). RESULTS: BWT (P<0.001) and LLD (P<0.001) scores were higher in the childhood asthma group than in the control group, but BD scores were not. The EDN (r=0.405, P<0.05) and ECP (r=0.565, P<0.01) levels significantly correlated with BWT scores, but not with LLD and BD in the childhood asthma group. The EDN (r=0.710, P<0.0001) and the ECP (r=0.580, P<0.0001) levels were significantly correlated with serum total eosinophil counts. CONCLUSION: The EDN and ECP levels were correlated with BWT scores on HRCT. These findings suggest that EDN and ECP may be valuable for quantifying airway thickening in children with asthma exacerbation.
Adult ; Asthma* ; Child* ; Dilatation ; Eosinophil Cationic Protein* ; Eosinophil-Derived Neurotoxin* ; Eosinophils* ; Humans ; Inflammation ; Lung

PURPOSE: Eosinophil is one of the important inflammatory cell involved in the airway inflammation in childhood asthma. It has been demonstrated that markers of eosinophil activation, including eosinophil cationic protein or eosinophil protein X(EPX), are increased in childhood asthma. Furthermore, they are related to disease activity and are assumed to be helpful in monitoring the treatment effect as urinary EPX(U-EPX) can be obtained easily and in a noninvasive way in children of all ages. METHODS: Twenty-five children(22 male and three female) aged 11.87+/-3.82 years with stable asthma were challenged with methacholine and urine was collected from each child during the following periods; before methacholine challenge test(MCT); 0-3 hr after the end of MCT; 4-7 hr after the end of MCT; and 8-24 hr after the end of MCT. Bronchial reactivity was determined by using Dosimeter(Jeager, Germany) with serially diluted methacholine from 0.05 to 25.0 mg. The FEV1 less than 80% of baseline value were classified into positive MCT. U-EPX was measured with a sensitive and specific radioimmunoassay(Pharmacia & Upjohn AB, Uppsala, Sweden). Results were expressed as microgramEPX/mmol creatinine. RESULTS: An early airway response after MCT was associated with an increase of U-EPX excretion for 0-3 hr after methacholine inhalation in comparison with beseline values. Most subjects showed a small increase in U-EPX excretion during late asthmatic response for 4-7 hr, which then decreased to normal level in 8-24 hr. Also, a tendency for a higher increase of U-EPX was associated with a lower threshold of methacholine challenge and a longer duration of asthma. CONCLUSION: Measurement of EPX in urine is a noninvasive and easy method to assess the severity of airway inflammation in asthmatic children. It may be a helpful index of the events underlying the airway inflammatory responses during nonspecific bronchial challenge, and in monitoring asthma management.
Asthma* ; Child* ; Creatinine ; Eosinophil Cationic Protein ; Eosinophil-Derived Neurotoxin* ; Eosinophils* ; Humans ; Inflammation ; Inhalation ; Male ; Methacholine Chloride*

PURPOSE: Short statue or obesity has been reported in asthmatic children, but the results are inconsistent. Recently eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN) levels has been known as important markers of airway inflammation and reflect asthma severity as well. The aim of this study is to evaluate the growth status and to analyze the possible relation with serum EDN and ECP levels. METHODS: A total of 90 children (57 boys and 33 girls, 4 to 16 years old) who had been admitted for bronchial asthma were included. To standardize the data for age and sex, standard deviation scores (SDS) were calculated for height and weight. Values less than -2 SDS below the mean were considered to be extremely low, -1 SDS to 1 SDS as normal, values higher than 2 SDS considered very high. Serum EDN and ECP levels were measured. RESULTS: The mean height SDS was 0.33+/-0.85 and weight SDS was 0.23+/-1.20. The prevalence of short stature was 2.2%, normal stature 75.5%, and tall stature 22.2%. The prevalence of underweight was 7.8%, normal weight 71.1%, and overweight 21.1%. Height SDS was negatively correlated with serum ECP (r=-0.27, P=0.01) and EDN (r=-0.27, P=0.009) and weight SDS was negatively correlated with serum ECP (r=-0.20, P=0.05). Height SDS were significantly lower in high ECP and EDN groups compared to normal ECP and EDN groups (P<0.01 and P<0.009, respectively). Weight SDS was lower in high ECP group compared to normal ECP group (P<0.05). CONCLUSION: Growth (height and weight) was inversely correlated with serum EDN and ECP levels. These results suggest that high ECP and EDN levels might be related with growth retardation of asthmatic children.
Asthma ; Child* ; Eosinophil Cationic Protein* ; Eosinophil-Derived Neurotoxin ; Eosinophils* ; Female ; Humans ; Inflammation ; Obesity ; Overweight ; Prevalence ; Thinness

Echinostoma cinetorchis is an oriental intestinal fluke causing significant pathological damage to the small intestine. The aim of this study was to determine a full-length cDNA sequence of E. cinetorchis endoribonuclease (RNase H; EcRNH) and to elucidate its molecular biological characters. EcRNH consisted of 308 amino acids and showed low similarity to endoribonucleases of other parasites (<40%). EcRNH had an active site centered on a putative DDEED motif instead of DEDD conserved in other species. A recombinant EcRNH produced as a soluble form in Escherichia coli showed enzymatic activity to cleave the 3′-O-P bond of RNA in a DNA-RNA duplex, producing 3′-hydroxyl and 5′-phosphate. These findings may contribute to develop antisense oligonucleotides which could damage echinostomes and other flukes.
Amino Acids ; Catalytic Domain ; DNA, Complementary ; Echinostoma* ; Endoribonucleases ; Escherichia coli ; Intestine, Small ; Oligonucleotides, Antisense ; Parasites ; Ribonuclease H* ; Ribonucleases* ; RNA ; Trematoda

<p>BACKGROUNDThe role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.p><p>METHODSTo detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.p><p>RESULTSIn 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).p><p>CONCLUSIONSDICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.p>
Blotting, Western ; DEAD-box RNA Helicases ; analysis ; genetics ; physiology ; Endoribonucleases ; analysis ; genetics ; physiology ; Epigenesis, Genetic ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Ribonuclease III ; Stomach Neoplasms ; chemistry ; etiology ; genetics

Asthma is associated with increased levels of eosinophils in tissues, body fluids, and bone marrow. Elevated levels of eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) have been noted in asthma patients. Higher levels of EDN and ECP are also associated with exacerbated asthmatic conditions. Thus, EDN, along with ECP, may aid the diagnosis and monitoring of asthma. Several groups have suggested that EDN is more useful than ECP in evaluating disease severity. This may partially be because of the recoverability of EDN (not sticky, 100% recovery rate), as ECP is a sticky and more highly charged protein. In terms of clinical utility, EDN level is a more accurate biomarker than ECP when analyzing the underlying pathophysiology of asthma. As a monitoring tool, EDN has shown good results in children with asthma as well as other allergic diseases. In children too young to fully participate in lung function tests, EDN levels may be useful as an alter native measurement of eosinophilic inflammation. EDN can also be used in adult patients and in multiple specimen types (e.g., serum, sputum, bronchoalveolar lavage fluid, and nasal lavage fluid). These results are repeatable and reproducible. In conclusion, EDN may be a novel biomarker for the diagnosis, treatment, and monitoring of asthma/allergic disease.
Adult ; Asthma ; Biomarkers ; Body Fluids ; Bone Marrow ; Bronchoalveolar Lavage Fluid ; Child ; Eosinophil Cationic Protein ; Eosinophil-Derived Neurotoxin ; Eosinophils ; Humans ; Inflammation ; Nasal Lavage ; Respiratory Function Tests ; Sputum

Wheezing is one of the characteristic symptoms of asthma, but all preschool children with wheezing are not diagnosed with asthma. Preschool children are not cooperative enough to participate in spirometry and invasive tests. Thus, there is no conventional method to diagnose asthma in preschool children. We reviewed studies on non-invasive biomarkers for assessing asthma in preschool children. Specimens that can be easily obtained by non-invasive methods are blood, exhaled breath and urine. Eosinophils, eosinophil cationic protein and eosinophil-derived neurotoxin (EDN) in blood are helpful in evaluating eosinophilic inflammation of the airways. Exhaled breath contains nitric oxide, volatile organic compounds, various cytokines and mediators as analytical components. Fraction of exhaled nitric oxide has been used to assess the degree of eosinophil inflammation and has been standardized in school-age children and adults, but not yet in preschool children. Exhaled breath condensate (EBC) pH and various cytokines/mediators that are detected in EBC seem to be promising biomarkers for assessing asthma, but need more standardization and validation. There are several biomarkers useful for assessing asthma, but none are ideal. Some biomarkers need standardized methods of obtaining samples from uncooperative preschool children for clinical use and require sufficient validation. Recently, another activated eosinophil marker, serum EDN, has shown promising results as a biomarker for recurrent wheezing and asthma in preschool children.
Adult ; Asthma* ; Biomarkers* ; Child ; Child, Preschool* ; Cytokines ; Eosinophil Cationic Protein ; Eosinophil-Derived Neurotoxin ; Eosinophils ; Humans ; Hydrogen-Ion Concentration ; Inflammation ; Methods ; Nitric Oxide ; Respiratory Sounds* ; Spirometry ; Volatile Organic Compounds

PURPOSE: Tumor necrosis factor (TNF)-alpha and eosinophilic inflammation have their role in asthma, but there were no studies on respiratory syncytial virus (RSV) bronchiolitis. The aim of our study was to investigate whether TNF-alpha has a role in eosinophilic inflammation of lower respiratory tract infections with RSV and has the correlation with other cytokines. METHODS: Fifty children with first RSV bronchiolitis (RSV group) and 18 healthy children without any respiratory symptom and sign (control group) were enrolled. Clinical data, such as eosinophil-derived neurotoxin (EDN), eosinophil cationic protein (ECP), were analyzed. We measured interleukin (IL)-5, IL-8, TNF-alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), interferon (IFN)-gamma, eotaxin, and regulated on activation, normal T cell expressed and secreted (RANTES) in nasal lavage fluid in both groups. RESULTS: Eotaxin, GM-CSF, IL-8, IFN-gamma and TNF-alpha were higher in the RSV group than the control group. TNF-alpha correlated with an eosinophil-active cytokine, GM-CSF (r=0.86, P<0.0001), IFN-gamma (r=0.90, P<0.0001), and with eosinophil-active C-C chemokines such as eotaxin (r=0.50, P<0.0001). TNF-alpha also correlated with proinflammatory cytokines such as IL-8 (r= 0.81, P<0.0001). CONCLUSION: TNF-alpha correlated with eosinophil-active chemokines and cytokines. Therefore, TNF-alpha may have a role in eosinophilic inflammation in children with RSV bronchiolitis.
Asthma ; Bronchiolitis ; Chemokines ; Chemokines, CC ; Child ; Cytokines ; Eosinophil Cationic Protein ; Eosinophil-Derived Neurotoxin ; Eosinophils ; Granulocyte-Macrophage Colony-Stimulating Factor ; Granulocytes ; Humans ; Inflammation ; Interferons ; Interleukin-8 ; Interleukins ; Nasal Lavage Fluid ; Respiratory Syncytial Viruses ; Respiratory Tract Infections ; Tumor Necrosis Factor-alpha

<p>OBJECTIVETo determine the effect of type I transmembrane protein (IRE1alpha) deletions on the cell cycle of human periodontal ligament fibroblasts (hPDLFs) cells.p><p>METHODSBased on the IRE1alpha deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IREla (pD-Kinase, pD-Rnase). The full-length model and two mutant eukaryotic expression vectors were transfected into hPDLFs cells. Western blot analysis was performed to identify the expression in the cells. The changes in the cell cycle of hPDLFS cells were detected by flow cytometry (FCM).p><p>RESULTSThe two deletion mutants of IRE1alpha with eukaryotic expression vectors were successfully constructed and correctly expressed in hPDLFs cells based on Western blot analysis. Under stress conditions, the FCM assay showed that cell percentage of S phases increased, whereas that of G1 phases decreased in the IRE1alpha group (P < 0.05) compared with the control group of tunicamycin (TM) treatment. Moreover, the cell percentage of the S phases decreased, whereas that of the G1 phases increased in the D-Rnase group (P < 0.05) compared with the control. The deletion mutant D-Kinase had no influence on hPDLFS cell proliferation and cycle (P>0.05).p><p>CONCLUSIONUnder stress conditions, IRE1alpha can improve the cell cycle of hPDLFs cells from the G1 to the S phase. The deletion mutant D-Rnase cause hPDLFs cell growth arrest at the G1 phase, whereas deletion mutant D-Kinase has no significant effect.p>
Cell Cycle ; Cell Proliferation ; Endoribonucleases ; Fibroblasts ; Humans ; Periodontal Ligament ; Protein-Serine-Threonine Kinases ; Transfection