| Ⅰ Expression of Linc00324 in Rheumatoid Arthritis ObjectiveTo detect the expression level of linc00324 in rheumatoid arthritis patients,and the correlation with clinically relevant indexes was analyzed to provide new ideas for the diagnosis and treatment of rheumatoid arthritis.Methods1.Samples of peripheral blood were collected from 42 clinical patients and27 healthy donors.2.Total RNA was extracted from the isolated peripheral blood leukocytes,and then was reversely transcribed into cDNA.3.Specific primers were designed according to the sequence of linc00324.The relative expression level of linc00324 was analyzed by real-time fluorescence quantitative PCR.4.The relationship between the expression level of linc00324 and rheumatoid arthritis was analyzed by stastical analysis,the clinical factors of patients,including age,gender,white blood cell count,cell type and phenotype,medication,immunoglobulin,complement,rheumatoid factor,erythrocytesedimentation rate,HLA-B27,Anti-cyclic citrullin polypeptide antibodies,anti-keratin antibodies,C-reactive protein,anti-nuclear antibodies,etc.5.The diagnostic and prognostic value of linc00324 was evaluated by ROC cruve.Results1.RNA quality analysis: The RNA concentration was above 30μg/ml and the OD260/280 was between 1.9 and 2.1.Agarose gel electrophoresis showed no significant degradation of RNA.2.The RNA was reverse transcribed into cDNA.There was only one peak in the dissociation curve obtained by fluorescence quantitative PCR amplification,indicating that the amplification product was homogeneous without non-specific amplification or primer dimer.3.The expression of linc00324 in leukocytes in patients with rheumatoid arthritis was significantly higher than that in the normal group(t=5.19,P=0.012).4.After the medical treatment,the expression of linc00324 in patients with rheumatoid arthritis was significantly lower than before treatment(t=6.46,P=0.00)..5.Linc00324 expression was positively correlated with leukocyte count(r=0.929,P=0.00),CD4 cell count(r=0.906,P=0.00),and positively correlated with CD4/CD8 ratio(r=0.541,P= 0.046),but no correlation with CD8 cell count.6.The expression of linc00324 was positively correlated with rheumatoid factor(r=0.444,P=0.034)and negatively correlated with anti-cyclic citrulline antibody(r=0.508,P=0.031),but no correlation with age,sex,IgG,Ig A,IgM,complement C3,complement C4,erythrocyte sedimentation rate,HLA-B27,C-reactive protein,anti-keratin antibody AKA and anti-nuclear antibody ANA.7.The area under ROC curve of linc00324 and rheumatoid factor were0.772 and 0.893 respectively.Z-test showed that there is no significant difference between them(P=0.064).ConclusionThe expression of linc00324 in patients with rheumatoid arthritis was significantly upregulated,which was significantly correlated with some clinical factors.These results suggest that linc00324 may plays an important role in the occurrence and development of rheumatoid arthritis and may provides a new idea for diagnosis,targeted therapy and prognosis of rheumatoid arthritis..Ⅱ Expression of Linc00324 in Systemic Lupus ErythematosusObjective To detect the expression level of linc00324 in systemic lupus erythematosus,and explore the relationship between linc00324 and the clinical signification of systemic lupus erythematosus..Methods1.Samples of peripheral blood were collected from 56 clinical patients and27 healthy donors2.Total RNA was extracted from the isolated peripheral blood leukocytes,and then was reversely transcribed into c DNA.3.Specific primers were designed according to the sequence of linc00324.,The relative expression level of linc00324 was analyzed by real-time fluorescence quantitative PCR.4.The relationship between the expression level of linc00324 and systemic lupus erythematosus was analyzed by stastical analysis,the clinical factors of patients,including age,gender,white blood cell count,cell type and phenotype,medication,immunoglobulin,complement,rheumatoid factor,erythrocyte sedimentation rate,C-reactive protein,anti-ds DNA antibodies,anti-nuclear antibodies,anti-keratin antibodies,anti-cyclic proline antibodies,etc.5.The diagnostic and prognostic value of linc00324 was evaluated by ROC cruve.Results1.RNA quality analysis: the RNA concentration was above 30μg/ml,and the ratio of OD260/280 was between 1.9 and 2.1.Agarose gel electrophoresis showed no significant degradation of RNA.2.The RNA was reverse transcribed into c DNA.There was only one peak in the dissociation curve obtained by fluorescence quantitative PCR amplification,indicating that the amplification product was homogeneous without non-specific amplification or primer dimer.3.The expression of linc00324 in patients with systemic lupus erythematosus was significantly higher than that in the normal group(t=7.21,P=0.00).4.After the medical treatment,the expression of linc00324 in patients with systemic lupus erythematosus was significantly lower than before treatment(t=6.51,P=0.00).5.Linc00324 expression was positively correlated with leukocyte count(r=0.661,P=0.00),CD4 cell count(r=0.895,P=0.00),and CD8 cell count(r=0.567,P=0.001),but no correlation with CD4/CD8 ratio(P>0.05).6.The expression of linc00324 was positively correlated with anti-dsDNA antibody(r=0.775,P=0.00),Ig G(r=0.453,P=0.004),and complement C3(r=0.392,P= 0.001),negatively correlated with anti-cyclic proline antibody(r=-0.565,P=0.035),but no correlation with age,sex,Ig A,Ig M,C-reactive protein,complement C4,erythrocyte sedimentation rate,rheumatoid factor,anti-keratin antibody AKA,and antinuclear antibody ANA.7.The area under ROC curve of linc00324 and anti-dsDNA antibodies was0.739 and 0.897,respectively.Z-test showed that there was no significant difference between them(P=0.854).ConclusionThe expression of linc00324 in patients with systemic lupus erythematosus was significantly upregulated,which was significantly correlated with some clinical factors.These results suggest that linc00324 may plays an important role in the occurrence and development of rheumatoid arthritis and may provides a new idea for diagnosis,targeted therapy and prognosis of systemic lupus erythematosus. |
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