ABSTRACT
Lipopolysaccharide (LPS) is the major constituents of the outer membrane of Gram-negative bacteria. LPS recognition and signal transmission are key events in the host defense reaction towards Gram-negative bacteria and are associated with many disorders. Multiple signaling pathways are involved in the response to LPS. With the help of LPS-binding protein and CD14, TLR4 binds with LPS, then recruits myeloid differentiation factor 88 and IL-1 receptor-associated kinase, and further phosphorylates and activates TNF receptor associated factor 6 (TRAF6). The activated TRAF6 leads to the activation of transcription factor NF-kappaB and MAP kinase's pathways that involves in LPS-induced cellular responses and the production of proinflammatory cytokines such as TNF-alpha, IL-6 and IL8.
Subject(s)
Acute-Phase Proteins , Metabolism , Carrier Proteins , Metabolism , Gram-Negative Bacteria , Chemistry , Lipopolysaccharide Receptors , Metabolism , Lipopolysaccharides , Pharmacology , Membrane Glycoproteins , Metabolism , Mitogen-Activated Protein Kinase Kinases , Metabolism , NF-kappa B , Physiology , Signal TransductionABSTRACT
OBJECTIVE@#To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS).@*METHODS@#Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope.@*RESULTS@#SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells.@*CONCLUSION@#SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.
Subject(s)
Humans , Cell Line, Tumor , Glycoproteins , Pharmacology , Membrane Proteins , Chemistry , Nasopharyngeal Neoplasms , Genetics , Pathology , Phosphoproteins , Pharmacology , Pseudomonas aeruginosa , Respiratory Mucosa , Chemistry , Allergy and Immunology , Respiratory System , Chemistry , Allergy and Immunology , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.</p><p><b>METHODS</b>Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.</p><p><b>RESULTS</b>D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).</p><p><b>CONCLUSION</b>These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Alleles , China , F-Box Proteins , Genetics , Gene Frequency , Genetic Linkage , Genetic Predisposition to Disease , Genetics , Microsatellite Repeats , Genetics , Nasopharyngeal Neoplasms , Genetics , PedigreeABSTRACT
<p><b>OBJECTIVE</b>To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area.</p><p><b>METHODS</b>Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.</p><p><b>RESULTS</b>Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black.</p><p><b>CONCLUSION</b>These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.</p>