ABSTRACT
<p><b>OBJECTIVE</b>To increase the recombinant adenovirus vector mediated human rotavirus VP6 gene expression through coden optimization.</p><p><b>METHODS</b>We have artificially synthesized VP6 gene of group A human rotavirus according to the human biased codon. The modified gene was transfected into 293 cells using adenovirus vector and the gene product, the respective protein was produced. The expression level of optimized gene and wild type gene was detected by Immunofluorescence (IF) and Western Blot.</p><p><b>RESULT</b>A remarkable increase of the expression level of optimized VP6 gene in comparison with the wild-type control.</p><p><b>CONCLUSION</b>The coden optimization indeed help increasing the recombinant adenovirus mediated human rotavirus gene expression, which indicated the potential application of such recombinant adenoviruses in the development of adenoviral-vectored rotavirus vaccines.</p>
Subject(s)
Humans , Adenoviridae , Genetics , Antigens, Viral , Genetics , Capsid Proteins , Genetics , Codon , Recombinant ProteinsABSTRACT
<p><b>BACKGROUND</b>Matrix metalloproteinase-1 (MMP-1) plays an important role in atherosclerosis. This study was to examine expression of MMP-1 mRNA in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and to explore its relationship with atherosclerosis in SLE.</p><p><b>METHODS</b>Fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of MMP-1 mRNA in PBMCs in 80 SLE patients, including 39 prone to atherosclerosis (Group A) and 41 unprone to atherosclerosis (Group B). Meanwhile, 30 patients who were free of cardiovascular diseases and 30 healthy individuals were selected as disease and normal control group (Groups C and D). The changes of MMP-1 gene expression were analyzed by differences of cycle threshold (DeltaCt), with the following formula: DeltaCt = Ct(target) gene - Ct(reference) gene.</p><p><b>RESULTS</b>The expression level of MMP-1 mRNA in Group A was significantly higher than that of group B (DeltaCt = 8.64 +/- 2.43 vs DeltaCt = 12.09 +/- 2.26, t = 6.588, P < 0.01). The expression level of MMP-1 mRNA of SLE patients was significantly higher than that of Group C (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.29 +/- 2.51, t = 3.135, P < 0.01) and Group D (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.48 +/- 1.69, t = 3.675, P < 0.01).</p><p><b>CONCLUSIONS</b>In comparison to disease and control group, expression of MMP-1 mRNA in PBMCs of SLE patients was significantly elevated, and significant difference of MMP-1 mRNA expression was also found between SLE patients prone and unprone to atherosclerosis, indicating that expression of MMP-1 mRNA may be correlated with the pathogenesis and activity of atherosclerosis in SLE.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Atherosclerosis , Genetics , Leukocytes, Mononuclear , Metabolism , Lupus Erythematosus, Systemic , Genetics , Matrix Metalloproteinase 1 , Genetics , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To compare the three Anti-dsDNA antibody detecting test (IIF, ELISA, Farr) with 200 serum samples to evaluate which one has higher sensitivity and specificity.</p><p><b>METHODS</b>200 serum samples including 120 serum samples of SLE, 20 serum samples of rheumatoid arthritis, 20 serum samples of MCTD, 20 serum samples of SS, 20 serum samples of PSS and 50 serum samples of healthy measured by IIF, Farr and ELISA.</p><p><b>RESULTS</b>Detection the Anti-dsDNA antibody of the serum sample with the methods of IIF, ELISA and Farr. The positive percentage of Anti-dsDNA in SLE is 25%, 32% and 32%, while in RA is 0, 0 and 0; in PSS is 0, 0 and 5%; in SS is 0, 0 and 0; in healthy is 0, 0 and 0.</p><p><b>CONCLUSION</b>Detection the Anti-dsDNA antibody with two method in the same time, especially with IIF and ELISA, will heighten the positive rate than with single method and will be helpful for the diagnosis of SLE.</p>
Subject(s)
Humans , Antibodies, Antinuclear , Allergy and Immunology , Case-Control Studies , DNA , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Immunologic Tests , Methods , Lupus Erythematosus, Systemic , Diagnosis , Allergy and Immunology , RadioimmunoassayABSTRACT
Objective To investigate the mRNA level of glucose-6-phosphate isomerase(GPI)ex- pression in patients with rheumatoid arthritis(RA).Method Reverse transcription-polymerase chain reaction (RT-PCR)was applied to semiquantitatively analyze GPI mRNA expression in the peripheral blood mononu- clear cells(PBMCs)of 30 active RA patients,30 stable RA patients,30 patients with other rheumatic disease and 30 healthy subjects.Results There was statistically significant differences between patients with RA and other rheumatic diseases(P