ABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>There 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.</p><p><b>CONCLUSION</b>CC16 gene may be not a susceptibility gene of asthmatic patients of Han population in southwest China.</p>
Subject(s)
Adult , Aged , Humans , Middle Aged , Alleles , Asthma , Genetics , China , Ethnology , Genetic Predisposition to Disease , Genotype , Mutation , Proteins , Genetics , UteroglobinABSTRACT
<p><b>OBJECTIVE</b>Screening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC).</p><p><b>METHODS</b>The differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis.</p><p><b>RESULTS</b>Thirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC.</p><p><b>CONCLUSION</b>The subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC.</p>
Subject(s)
Humans , Base Sequence , Carcinoma, Hepatocellular , Genetics , Cloning, Molecular , DNA, Complementary , Chemistry , Genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Gene Library , Liver Neoplasms , Genetics , Molecular Sequence Data , Sequence Analysis, DNAABSTRACT
Objective To study the effect of IFN-γ inhalation via aerosol on cytokines of the immunocompromised rats. Methods Immunocomprised rat model was established with cortisol acetate injection for 14 d and then Candida albicans fluid was injected by tracheal for establishing am immuno comprised with pulmonary infection model. IFN-γ was inhaled with aerosol 1 d before the bacterium injection and then for 1, 3 and 7 d respectively. The activity of TNF-α, and the levels of IL-1β and IL-6 in the supernatant of the cultured alveolar macrophage(AM), the activity of IFN-γ and TNF-α in bronchial alveolar lavage fluid (BALF), the expressions of IFN-γ,TNF-α, IL-1β, and IL-6 of the lung tissues, the level of IFN-γ,IL-1β, and IL-6 in the serum were investigated. Results The activity of TNF-α, and the levels of IL-1β and IL-6 in the culture supernatant of the AM of the rats treated with IFN-γ were significantly higher than those of the control. The activity of IFN-γ and TNF-α in BALF was higher in the IFN-γ inhaled rats than in the control (except the activity of TNF-α on the 7th day). The expressions of IFN-γ and IL-1β in lung tissues was higher in the rats treated with IFN-γ than in the control. The expression of TNF-α in the rats treated with IFN-γ was less than that in the control rats. The expression of IL-6 had no difference between 2 groups. And no difference was found in the activity of IFN-γ, and the levels of IL-1β and IL-6 in the serum between 2 groups(except IL-1β on the 3rd day). Conclusion Administration of IFN-γ via aerosol can obviously increase the activity or levels of some cytokines in the lung of the immunocompromised rats, but has no effect on them in serum of the immunocompromised rats.
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Objective To establish a multidrug resistance cell line from human lung adenocarcinoma. Methods The human lung adenocarcinoma cell line SPC-A-1 was exposed to cisplatin of a high and then increasing concentration for 192 d to establish multidrug resistance cell line (SPC-A-1/CDDP). The relative resistance was tested with MTT assay. The morphology of the cells was observed with transmission and scanning electron microscopy and the chromosome of them was analyzed with Giemsa stained specimens. Results The resistance index of SPC-A-1/CDDP cells to cisplatin was 11.2 and the cells showed various cross-resistance to 5-Fu, doxorubicin, mitomycin, vincristine and etoposide, but not to hydroxycamptothecine. Electron microscopy showed the cells with irregular and enlarged nuclei and abundant microvilli. Conclusion A multidrug resistance cell line (SPC-A-1/CDDP) from human lung adenocarcinoma is established. It can be used to downstream experiment.
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Objective To study the effect of transfecting anti-sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods MCT1 antisense gene recombinant vector pLXSN-MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection（P<0.001）. Conclusion MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.
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Objective To clone the partial positive regulatory fragment of Na+/H+ exchanger-1 (NHE-1) gene from human lung cancer cells. Methods After BamHⅠ and EcoRⅠ cut sites were added to the 5' ends of the upstream and downstream primers respectively, the partial positive regulatory sequence of NHE-1 gene was cloned with the length of 170 bp from genomic DNA of lung cancer cell line A549 cells with PCR method. The cloned fragment was ligated to plasmid pUC18. Finally, the constructed recombinant was identified with enzyme cut, PCR and DNA sequencing. Results The cloned fragment was about 170 bp in size and successfully ligated to pUC18 with identifiation of double enzyme cut and PCR. DNA sequencing approved that the fragment cloned was objective one with 168 bp in length. Compared with the reported sequence, two t were lost. Conclusion The positive regulatory fragment of NHE-1 gene from human lung cancer cells was successfully cloned.
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Objective To study the effect of IFN-γ inhalation via aerosol on cytokines of the immunocompromised rats. Methods Immunocomprised rat model was established with cortisol acetate injection for 14 d and then Candida albicans fluid was injected by tracheal for establishing am immuno comprised with pulmonary infection model. IFN-γ was inhaled with aerosol 1 d before the bacterium injection and then for 1, 3 and 7 d respectively. The activity of TNF-α, and the levels of IL-1β and IL-6 in the supernatant of the cultured alveolar macrophage(AM), the activity of IFN-γ and TNF-α in bronchial alveolar lavage fluid (BALF), the expressions of IFN-γ,TNF-α, IL-1β, and IL-6 of the lung tissues, the level of IFN-γ,IL-1β, and IL-6 in the serum were investigated. Results The activity of TNF-α, and the levels of IL-1β and IL-6 in the culture supernatant of the AM of the rats treated with IFN-γ were significantly higher than those of the control. The activity of IFN-γ and TNF-α in BALF was higher in the IFN-γ inhaled rats than in the control (except the activity of TNF-α on the 7th day). The expressions of IFN-γ and IL-1β in lung tissues was higher in the rats treated with IFN-γ than in the control. The expression of TNF-α in the rats treated with IFN-γ was less than that in the control rats. The expression of IL-6 had no difference between 2 groups. And no difference was found in the activity of IFN-γ, and the levels of IL-1β and IL-6 in the serum between 2 groups(except IL-1β on the 3rd day). Conclusion Administration of IFN-γ via aerosol can obviously increase the activity or levels of some cytokines in the lung of the immunocompromised rats, but has no effect on them in serum of the immunocompromised rats.
ABSTRACT
Objective To establish a multidrug resistance cell line from human lung adenocarcinoma. Methods The human lung adenocarcinoma cell line SPC-A-1 was exposed to cisplatin of a high and then increasing concentration for 192 d to establish multidrug resistance cell line (SPC-A-1/CDDP). The relative resistance was tested with MTT assay. The morphology of the cells was observed with transmission and scanning electron microscopy and the chromosome of them was analyzed with Giemsa stained specimens. Results The resistance index of SPC-A-1/CDDP cells to cisplatin was 11.2 and the cells showed various cross-resistance to 5-Fu, doxorubicin, mitomycin, vincristine and etoposide, but not to hydroxycamptothecine. Electron microscopy showed the cells with irregular and enlarged nuclei and abundant microvilli. Conclusion A multidrug resistance cell line (SPC-A-1/CDDP) from human lung adenocarcinoma is established. It can be used to downstream experiment.
ABSTRACT
Objective To study the effect of transfecting anti-sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods MCT1 antisense gene recombinant vector pLXSN-MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection（P<0.001）. Conclusion MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.
ABSTRACT
Objective To clone the partial positive regulatory fragment of Na+/H+ exchanger-1 (NHE-1) gene from human lung cancer cells. Methods After BamHⅠ and EcoRⅠ cut sites were added to the 5' ends of the upstream and downstream primers respectively, the partial positive regulatory sequence of NHE-1 gene was cloned with the length of 170 bp from genomic DNA of lung cancer cell line A549 cells with PCR method. The cloned fragment was ligated to plasmid pUC18. Finally, the constructed recombinant was identified with enzyme cut, PCR and DNA sequencing. Results The cloned fragment was about 170 bp in size and successfully ligated to pUC18 with identifiation of double enzyme cut and PCR. DNA sequencing approved that the fragment cloned was objective one with 168 bp in length. Compared with the reported sequence, two t were lost. Conclusion The positive regulatory fragment of NHE-1 gene from human lung cancer cells was successfully cloned.