No abstract available.
Uteroglobin*

A mammalian expression plasmid pcDNA3. 1-hCC10 was constructed and identified, then CC10 protein expression in A549 lung cancer cell line was detected. A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by using RT-PCR and cloned into expression plasmid cDNA3. 1, and the recombinant plasmid was identified by employing double digestion restriction enzymes Hind III and BamH I and the cDNA sequence was assayed by the Sanger dideoxy-mediated chain termination method. The segment was then transfected into the A549 lung cancer cell line. The protein expression of CC10 was detected by immunofluorescence and Western blot. Our results showed that the cDNA fragment included the entire coding region (273 bp). The recombinant eukaryotic cell expression vector of pcDNA3. 1-hCC10 was successfully constructed, and the sequence of the insert was identical to the published sequence. A549 cells line transfected with the pcDNA3. 1-hCC10 expressed high level of CC10 protein. The recombinant plasmid cDNA3. 1-hCC10 may serve as an effective tool for the study of tumorogenesis and tumor treatment.
Adenocarcinoma/*metabolism ; Adenocarcinoma/pathology ; Cell Line, Tumor ; Genetic Vectors ; Lung Neoplasms/*metabolism ; Lung Neoplasms/pathology ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Transfection ; Uteroglobin/biosynthesis ; Uteroglobin/*genetics

BACKGROUND: Uteroglobin is a protein produced by the normal bronchial epithelium and its expression level is lower in non-small cell lung cancer tissues and cell lines. It mainly functions as an anti-inflammatory, and when it is overexpressed in cancer cells, the neoplastic phenotype is antagonized. cPLA2 and COX-2, which are also associated with inflammation, were reported to be related to cancer. The relationship between cPLA2, COX-2 and uteroglobin is unclear. The relationship between uteroglobin and ERK, which is related to cell growth, is also not unclear. This study investigated the changes in the cPLA2 and COX-2 expression levels and the ERK activities after the overexpression of uteroglobin in non-small cell lung cancer cell lines. METHODS: The A549 and NCI-H460 cell lines were infected by adenovirus-null and adenovirus- uteroglobin. The cChange in the cPLA2, COX-2 expression level and ERK activity after uteroglobin overexpression was measured by Western blot. The change in MMP activity was measured by zymography. RESULTS: Western blot revealed decreased expression levels of cPLA2, and COX-2, and increased pERK levels in nonsmall cell lung cancer cells after uteroglobin overexpression. Zymography revealed no changes in the MMP-2 activity and lower MMP-9 activity. U0126, which is a specific inhibitor of ERK-activating kinase MEK-1/-2, prevented the decrease in the MMP-9 activity CONCLUSIONS: A decrease in cPLA2 expression, COX-2 expression, MMP-9 activity and a increase in ERK activity may be related to the anticancer effects of uteroglobin in nonsmall cell lung cancer cells.
Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; Cell Line ; Epithelium ; Inflammation ; Lung Neoplasms ; Phenotype ; Phosphotransferases ; Uteroglobin*

Glomerulonephritis(GN) is characterized by cognate immune responses against self or non-self antigen. It is suggested that the crescentic GN is a manifestation of cell-mediated immune response akin to delayed type hypersensitivity. Uteroglobin(UG) is a steroid-dependent, immunomodulatory, and cytokine-like protein. It was reported that UG prevented fibronectin(Fn) deposition in the glomeruli of normal mice to form Fn-UG heterodimers that competed with Fn self-aggregation. We hypothesized that UG would prevent the development of experimental GN induced by anti-glomerular basement membrane globulin(anti-GBM Ab) in mice through immunomodulatory properties. GN was induced by intravenous injection of 4.5 mg rabbit anti-GBM Ab to mice(C57BL/6). Renal injury was evaluated at 7, 14, and 21 days thereafter. UG-treated mice(n=10) were received for 3 days(0.5 mg/mouse/day) beginning 1 hour after anti-GBM Ab injection. Also, disease-control mice(n=10) were received PBS for 3 days after anti-GBM Ab. Proteinuria was significantly reduced in the mice treated with UG when compared with the disease-control mice after 7 and 14 days of anti-GBM Ab injection. The amount of proteinuria was similar between UG treated and normal control mice. The mesangial matrix expansion and cellular crescent were markedly attenuated by the injection of UG. The proliferative responses of mesangial cells(C57BL/6) to LPS were blunted with the addition of UG in dose-dependent manner. In this study, we revealed the preventive effects of UG in the experimental model of glomerulonephritis. This result in turn could provide the basis for the treatment of human disease such as chronic glomerulonephritis.
Animals ; Basement Membrane ; Glomerulonephritis* ; Humans ; Hypersensitivity ; Injections, Intravenous ; Mice ; Models, Theoretical ; Proteinuria ; Uteroglobin*

Adolescent ; Asthma ; blood ; Bronchitis ; blood ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Uteroglobin ; blood

OBJECTIVETo study the expression of serum Clara cell secretory protein 10 (CC10) and total IgE concentration in wheezing children under 5 years old.

METHODSFifty-nine children with recurrent wheezing under 5 years old were classified into two groups: wheezing group 1 with atopic high risks (n=33) and wheezing group 2 without atopic high risks (n=26). Twenty-three children without infectious diseases served as a control group. Serum levels of CC10 and IgE were measured using a solid-phase sandwich ELISA.

RESULTSThe serum levels of CC10 in wheezing group 1 (3.95 ± 1.26 ng/mL) and wheezing group 2 (5.41 ± 1.64 ng/mL) were significantly lower than those in the control group (8.72 ± 2.23 ng/mL; P<0.01). The wheezing group 1 showed more decreased serum levels of CC10 compared with wheezing group 2 (P<0.05). The serum IgE levels in wheezing group 1 were significantly higher than those in wheezing group 2 and the control group (P<0.05). There were no significant differences in serum IgE levels between the wheezing group 2 and control group. There was a negative correlation between serum levels of CC10 and IgE in wheezing group 1 (r=-0.912, P < 0.01).

CONCLUSIONSSerum CC10 levels decrease remarkably in wheezing children, and more significant decrease is noted in patients with atopic high risks. Serum CC10 levels are negatively correlated to serum IgE levels in patients with atopic high risks.

Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Male ; Respiratory Sounds ; immunology ; Uteroglobin ; blood

PURPOSE: Uteroglobin (UG), steroid inducible cytokine-like protein, has potent anti-inflammatory and immunomodulatory action. It is secreted by the mucosal epithelia of virtually all mammals. The aims of this study were to investigate the efficacy of recombinant adenovirus carrying uteroglobin (AdCMV-UG) in prevention and treatment of glomerulonephritis (GN) in mice. METHODS: The AdCMV-UG was created by inserting the uteroglobin cDNA into the pAdTrack- CMV vector and was transfected into the 293 cells through liposome mediated vehicles. AdCMV-UG was injected direct to the both kidneys of 20 mice. In control groups (disease controls), 13 mice received adenoviral vector with GFP and another 11 mice received PBS only. After 5days of viral injection, GN was induced by repetitive intravenous injection of 3.0 mg rabbit anti-GBM Ab to the pretreated mice (C57/B6). Histological and biochemical changes were evaluated 7 and 14 days after injection of anti-GBM Ab. RESULTS: UG was expressed in the renal tissues and mesangial cells infected with the infection of AdCMV-UG. Pretreatment with AdCMV-UG attenuated the cellular crescent formation 7 days after induction of GN when compared to AdCMV-GFP, PBS only. We also observed reduced mesangial matrix expansion in mice treated with adenovirus carrying UG. Proteinuria was significantly reduced in the mice treated with adenovirus carrying UG when compared with disease control mice (AdCMV-UG 102.2+/-20.97, AdCMV-GFP 170.6+/-41.77, and PBS 169.8+/-55.67, respectively p<0.05 mg/mg). However, at 14 days after anti-GBM Ab injection (total 19 days), there was no significant difference in the amounts of prot einuria and morphologic findings between pretreated and disease control groups. CONCLUSION: Adenoviral mediated gene transfer is an effective way of gene delivery. Locally expressed uteroglobin attenuated the severity of glomerulonephritis induced by anti-GBM antibody, although it was transient. Gene therapy using uteroglobin may be constituted for the treatment of human diseases such as chronic GN.
Adenoviridae ; Animals ; DNA, Complementary ; Genetic Therapy* ; Glomerulonephritis* ; Humans ; Injections, Intravenous ; Kidney ; Liposomes ; Mammals ; Mesangial Cells ; Mice* ; Proteinuria ; Uteroglobin*

OBJECTIVETo investigate the possible association between polymorphism of CC16 gene exon 1 and asthma, the genotype and allele frequencies of CC16 gene exon 1 in the asthmatic patients of Han population in southwest China were analyzed.

METHODSThe authors determined the genotypes of CC16 gene exon 1 with polymerase chain reaction technique and restricted enzyme analysis, and then compared the genotype and allele frequencies of the gene of the asthmatic group with those of the healthy control group.

RESULTSThere was no significant difference in genotype and allele frequencies of CC16 gene between the asthmatic group and control group. There was no association between the genotype and allele frequencies of gene and the severity of asthma.

CONCLUSIONCC16 gene may be not a susceptibility gene of asthmatic patients of Han population in southwest China.

Adult ; Aged ; Alleles ; Asthma ; genetics ; China ; ethnology ; Genetic Predisposition to Disease ; Genotype ; Humans ; Middle Aged ; Mutation ; Proteins ; genetics ; Uteroglobin

The present study investigated the effect of antiflammin-1 (AF-1) on LPS-induced IL-10 secretion from RAW264.7 cells through uteroglobin-binding protein (UGBP). Cultured RAW264.7 cells, a murine monocyte-macrophage cell line, were divided as following: control group, LPS group (1 µg/mL LPS), AF-1 group (100 μmol/L AF-1), LPS+AF-1 group (2 h of 100 μmol/L AF-1 pretreatment before LPS addition), and LPS+AF-1+anti-UGBP group (30 min of anti-UGBP antibody pretreatment before successive treatments with AF-1 and LPS). IL-10 concentration in the supernatants was detected by ELISA assay, and the level of IL-10 mRNA expression in macrophage was detected by using RT-PCR method. The results showed that AF-1 significantly increased LPS-induced IL-10 secretion in RAW264.7 cells in a dose dependent way, and up-regulated its mRNA level. Anti-UGBP antibody pretreatment attenuated the augmented effect of AF-1 on LPS-induced IL-10 secretion and gene expression. These results suggest that AF-1 promotes LPS-induced IL-10 secretion from macrophages, and this effect is mediated by UGBP.
Animals ; Cell Line ; Gene Expression ; Interleukin-10 ; metabolism ; Lipopolysaccharides ; Macrophages ; metabolism ; Mice ; Peptide Fragments ; metabolism ; RNA, Messenger ; Uteroglobin ; metabolism

OBJECTIVE: To investigate the expression of Clara cell 10-KDa protein (CC010) in sinonasal mucosa of murine bacterial chronic rhinosinusitis (BCRS) model. METHOD: A murine BCRS model was established by Streptococcus pneumoniae inoculation plus Merocel ostiomeatal obstruction. After 12 week's intervention, histological changes of sinonasal mucosa in BCRS model were examined by hematoxylin and eosin stain, periodic acid-schiff stain, and Masson-Trichrome stain. The mRNA and protein expression of CC10 in sinonasal mucosa were determined by reverse transcription polymerase chain reaction and immunohistochemistry methods. The number of CC10 positive cells in sinonasal epithelium was also counted. RESULT: In BCRS model group, polymorphonuclear neutrophils (PMN), subepithelial collagen deposition, goblet cells, and epithelial thickness were significantly increased, compared with control group (P<0.01). However, CC10 positive cells, CC10 mRNA and protein expression in sinonasal mucosa of BCRS model group were significantly decreased, compared with control group (P<0.01). Moreover, the number of CC10 positive cells was significantly negatively correlated with PMN (r=-0.734, P<0.01), subepithelial collagen deposition (r=-0.776, P<0.01), epithelial goblet cells (r=-0.841, P<0.01), and epithelial thickness (r=-0.805, P<0.01), respectively. CC10 average grayscale value was significantly positively correlated with PMN (r=0.771, P<0.01), subepithelial collagen deposition (r=0.802, P<0.01), epithelial goblet cells (r=0.887, P<0.01), and epithelial thickness (r=0.855, P<0.01), respectively. CONCLUSION The expression of CC10 is downregulated in sinonasal mucosa in BCRS model. As an important endogenous modulin, CC10 might play a crucial role in the pathogenesis of chronic rhinosinusitis.
Animals ; Chronic Disease ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Pneumococcal Infections ; metabolism ; Sinusitis ; metabolism ; microbiology ; Uteroglobin ; metabolism