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The research background rheumatoid arthritis (rheumatoid arthritis, RA) is a common chronic inflammatory joint tissue as the main manifestation of systemic autoimmune diseases. Lesions joints mainly as inflammatory cell infiltration, synovial hyperplasia, pannus formation, and the consequent damage of cartilage and bone. RA was global distribution, the incidence worldwide average of 1%, in our country the incidence rate of 0.26% to 0.5%, the male to female ratio of 1:3, and can occur at any age, the peak incidence of 45 to 50 years old. RA is one of the major disabling diseases, resulting in loss of joint function, bring tremendous pressure and burden to the society and the family. The complexity of the pathogenesis of RA, wherein the participation of a variety of immune cells and cytokines. RA occurs CD4 T cell activation and proliferation, secrete a variety of cytokines, such as interleukin (IL) -1 (IL-1) and tumor necrosis factor (TNF), followed by the macrophage cells, B cells, endothelial cells are activated, the activated B cells produce rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibody (anti-CCP). IL-1, TNF-α and other inflammatory or pro-inflammatory cytokines in the pathogenesis plays the role of the \Recently, studies have shown that the RNA-microRNA (miRNA, micro RNA), a class of small molecules participate in T and B lymphocyte differentiation, involved in the innate immune response and the adaptive immune response, may in autoimmune diseases such as RA play an important role in . miRNA is a class of a length of about 21 to 25 bases of non-protein-coding single-stranded RNA, small molecules widely exist in multi-cellular organisms and viruses in vivo, mainly through the nucleic acid sequence complementary to match specific binding to target mRNA, inhibition of target mRNA translation The procedure or degradation of the target mRNA, is a negative regulator role molecules. Predicted that about 30% of the gene coding sequence by the regulation of miRNAs. miR-146a and miR-16 is one of the two. Through the regulation of its target genes, miR-146a in RA, affect transcription factors (NF-κB) dependent secretion of cytokines such as IL-1β, TNF-a and other synovial cell proliferation, inflammatory response mediated RA patients . miR-16 rapid degradation of regulatory elements in the 3 'non-coding region containing AU, including TNF-a, IL-8, IL-6, inhibit excessive inflammatory response. Research miR-146a and miR-16 expression levels in the peripheral blood of patients with RA and the degree of clinical indicators of disease activity, helps to further explore its significance in the pathogenesis of RA and disease progression. RA belongs to the motherland's traditional medicine \The Arthralgia occurrence mainly due to the lack of righteousness, to feel the wind, cold, wet, heat caused by evil. The internal causes are the basis of ferrite weakness, lack of righteousness, Cou reason not close, the guard outside is not solid, internal factors caused Arthralgia, feel the evils, such as the muscles, joints, meridians Blockage form Arthralgia. Arthralgia treatment Qufeng basic Dafa governance to Qufeng, dehumidification, Shujin. Sinomenine preparation (Qingfengdian pain capsules) with expelling wind and dampness, warm and cold, the efficacy Huoxuetongluo, past pharmacological and clinical studies have shown to have good anti-inflammatory analgesic effect on acute and chronic inflammation. Methotrexate (MTX) is still the most widely used drugs RA to improve the condition, has anti-inflammatory and immunosuppressive effects. Combined treatment of active RA has a good effect. The study the sinomenine preparations (breeze pain capsules) in combination with MTX miR-146a and miR-16 expression levels contribute to the further explore sinomenine preparation (Qingfengdian pain capsules) joint mechanism of MTX in RA. Treatment is one of the basic characteristics of the traditional Chinese medicine, TCM establish governance law, basis contingent Prescription. Chinese medicine in the diagnosis and treatment of the disease process, Li-fa Recipe will stretch through in order to reflect the essence of traditional Chinese medicine, bridges and key card is the Li-fa Recipe stretch through. RA diagnostic studies at home and abroad a lot of Chinese medicine and more from the overall concept, diagnosis and treatment from a macro perspective, modern medicine more than to start from the laboratory tests, research from the microscopic point of view, both have their own strengths and have The short. The miRNAs with TCM-no reported. Explore the miRNAs RA TCM-related, all have an important objective research and guide clinical treatment of RA dialectical. Research purposes. Study in patients with RA peripheral blood of miR-146a, miR-16 expression analysis of miR-146a, miR-16 expression with RA disease activity relationship; 2. Observed sinomenine preparation (Qingfengdian pain capsules) joint MTX in RA peripheral blood of patients with miR-146a and miR-16 expression levels; 3. explore RA patients peripheral blood of miR-146a, miR-16 expression TCM-relations. Method 1. Study Nanfang Hospital, Southern Medical University Chinese rheumatology outpatient and ward RA patients diagnosed from June 2009 to March 2010, 40 cases, 8 males, female 32 cases, 24 cases of active patients were male 4 cases, 20 were females. Remission period 16 patients, 4 males and 12 females. The healthy control group of 16 healthy individuals for the same period in hospital outpatient, 4 males and 12 females. 2. The research method 2.1RA patients with peripheral blood of miR-146a, miR-16 expression and disease activity relationship miR-146a in peripheral blood mononuclear cells using real-time quantitative PCR assay RA patients, the relative expression of miR-16 to U6snRNA as an internal, 2 - △ △ CT analysis method for the relative quantification of the expression level of miR-146a, miR-16. Collect the number of swollen joints in patients with (Swollen0-28), tender joint count (Tender0-28), the overall health status score, rheumatoid factor (RF), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and other clinical data, analysis RA the peripheral blood of patients with miR-146a, miR-16 expression and disease activity index correlation. 2.2 the sinomenine preparations (breeze pain capsules) in combination with MTX miR-146a and miR-16 expression levels in peripheral blood of RA patients, 18 patients with active RA patients to of sinomenine preparations (breeze pain capsules) in combination with MTX 12 weeks of treatment, Real-time PCR assay RA patients before and after treatment, the expression level of miR-146a and miR-16 in peripheral blood. 2.3miR-146a, miR-16 expression and TCM type of relationship based on the guiding principles of the \16 cases of type. SPSS13.0 statistical software. Statistical processing of all data analysis. Measurement data (X ± S). Two sample measurement data were compared using independent sample t test, the difference before and after the group using paired t-test, multiple sets of samples were compared using One-way ANOVA, correlation analysis using Spearman correlation analysis or Person method, P ≤ 0.05 for difference statistics significance. Results 1. Relationship 1.1RA real-time quantitative PCR assay of miR-146a, miR-16 relative expression of miR-146a, miR-16 expression and disease activity in patients with peripheral blood mononuclear cell total RNA polyacrylamide gel electrophoresis 28S, 18S, 5S 3 with the UV spectrophotometer absorbance (A) 260/280 ratio at 1.8 to 2.0. Whether object miRNAs or internal reference U6snRNA of the amplification curve and dissolution curve is good, no primer dimers, no non-specific amplification. 1.2 miR-146a and miR-16 expression in peripheral blood mononuclear cells in patients with RA were significantly higher than the healthy control group (P = 0.004, P = 0.001). Mitigation activities group miR-146a expression levels above 1.3 RA group (P = 0.035) and the control group (P = 0.005), RA remission group of miR-146a expression levels compared to the control group, the difference was not statistically significant (P = 1.000) . Active RA group miR-16 expression levels higher than the remission group (P = 0.037) and the control group (P = 0.002), RA remission group of miR-16 expression levels compared with the control group showed no significant difference (P = 0.755). 1.4 RA expression levels of miR-146a in patients with swollen joint count (r = 0.354, P = 0.007), tender joint count (r = 0.352, P = 0.008), ESR (r = 0.541, P = 0.000), CRP (r = 0.402, P = 0.010), DAS28 (r = 0.417, P = 0.007) were positively correlated with RF (r = 0.132, P = 0.419) correlation. 1.5 RA miR-16 expression levels in patients with swollen joint count (r = 0.479, P = 0.000), tender joint count (r = 0.474, P = 0.000), ESR (r = 0.674, P = 0.000), CRP (r = 0.517, P = 0.001), DAS28 (r = 0.575, P = 0.000) were positively correlated with RF (r = 0.239, P = 0.137) correlation. 2 sinomenine preparation (breeze pain capsules) combined with MTX in RA peripheral blood of patients with miR-146a and miR-16 expression levels 2.1 sinomenine preparation (Qingfengdian pain capsules) in combination with MTX in patients with RA after 12 weeks of treatment in peripheral blood IniR -146a expression levels (P = 0.040). 2.2 the sinomenine preparations (breeze pain capsules) in combination with MTX after 12 weeks of treatment in peripheral blood of patients with RA miR-16 expression levels (P = 0.021). 3.miR-146a, 3.1 three groups of miR-146a expression level of miR-16 expression and TCM type of relationship with a significant difference (F = 19.911, P = 0.000). Rheumatoid folder stasis miR-146a expression levels higher than the liver and kidney loss type (P = 0.000) and the control group (P = 0.000), liver and kidney depleted miR-146a expression levels compared with the control group showed no significant difference (P = 0.852). 3.2 three sets of miR-16 expression levels are significantly different (F = 9.229, P = 0.000). Rheumatoid folder stasis miR-16 expression levels higher than the the hepatorenal loss type (P = 0.036) and the control group (P = 0.000), liver and kidney depleted miR-16 expression levels compared to the control group showed no significant difference (P = 0.827). 3.3 rheumatoid stasis RA patients ESR and CRP above the liver and kidney loss type (P = 0.001, P = 0.020), a statistically significant difference. 3.4 Rheumatoid stasis RA patients is lower than RF the hepatorenal loss type, the difference was statistically significant (P = 0.014); rheumatoid to folders Blood RA patients with anti-CCP antibody levels in liver and kidney loss in RA patients compared, the difference was not statistically significance (P = 0.145). The conclusion 1.RA peripheral blood of patients: miR-146a, miR-16 expression levels are raised, may be related to the pathogenesis of RA. 2.miR-146a, miR-16 expression levels and swollen joint count, tender joint count, ESR, CRP, DAS28 was positively correlated with no correlation with RF; detection of miR-146a, miR-16 expression levels may be monitoring RA valid indicators of disease activity. 3. Preparation of Sinomenine (breeze pain capsules) Joint MTX treatment expression levels of miR-146a, miR-16 down, may determine the efficacy of some significance. MiR-146a, miR-16, ESR, CRP, RF difference between significant 4.RA rheumatoid stasis, liver and kidney depleted prompt above indicators may be related with the TCM syndromes, TCM can provide a certain objective study of basis.
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