ABSTRACT
Objective The HOTAIR gene is closely related to pannus formation in rheumatoid arthritis (RA). This study aimed to construct and screen fibroblast-like synoviocytes in human RA (HFLS-RA) stably overexpressing lncRNA HOTAIR, and to pave the way for further study of the role of lncRNA HOTAIR in the pathogenesis of RA. Methods LncRNA HOTAIR was cloned and linked to the PMT406 vector digested by BamHI-HF-HF and XhoI. The constructed plasmids were sequenced, identified and then transfected into 293T cells to pack lentivirus. The HFLS-RA cells were infected with the recombinant and empty vector lentiviruses, and purinomycin was employed to screen the lncRNA HOTAIR-overexpressed and control cell lines. The total RNA was extracted from the blank, negatively transfected and overexpressed cells by Trizol, and the cDNA obtained by reverse transcription was amplified by qPCR, followed by determination of the expression of lncRNA HOTAIR. Results The relative expression of lncRNA HOTAIR was significantly higher in the overexpression group than in the blank control and negative transfection groups (30.329 ± 3.860 vs 1.001 ± 0.048 and 0.892 ± 0.247, P 0.05). Conclusion The HFLS-RA cell line stably overexpressing lncRNA HOTAIR was successfully constructed, which has provided some experimental evidence for further investigation of the role of lncRNA HOTAIR in the pathogenesis of RA.
ABSTRACT
<p><b>OBJECTIVE</b>To assess the short-term and long-term therapeutic effect of warming needle moxibustion for treatment of knee osteoarthritis.</p><p><b>METHODS</b>Ninety cases were randomly divided into a warming needle moxibustion group, a western medicine group and a waiting group, 30 cases in each group. The warming needle moxibustion group was treated with warming needle moxibustion on Xuehai (SP 10), Dubi (ST 35) and Zusanli (ST 36), etc.; the western medicine group was treated with oral administration of Ibuprofen sustained release capsule; the waiting group did not receive any treatment. Western Ontario and McMaster University Osteoarthritis Index (WOMAC) and Chinese version of SF-16 were used to assess the therapeutic effect before treatment, after treatment and 10 weeks after treatment.</p><p><b>RESULTS</b>After treatment for 2 weeks, the total effective rate was 86.7% in the warming needle moxibustion group, 90.0% in the western medicine group, and 60.0% in the waiting group, both the warming needle moxibustion group and the western medicine group being significantly better than the waiting group (both P<0.05) and with no significant difference between the warming needle moxibustion group and the western medicine group (P>0.05); 10 weeks after treatment, the total effective rate of 83.3% in the warming needle moxibustion group was better than that of 60.0% in the western medicine group (P<0.05). There were no significant differences in comparison of WOMAC and SF-16 scores after treatment between the warming needle moxibustion group and the western medicine group, and the warming needle moxibustion group in improvement of these scores was superior to the western medicine group (P<0.01, P<0.05).</p><p><b>CONCLUSION</b>The therapy of warming needle moxibustion is effective for treatment of knee osteoarthritis, and the short-term therapeutic effect is same as that of oral administration of Ibuprofen and the long-term therapeutic effect is better than that of Ibuprofen.</p>