Objective： To compare differences of acupotomy loosing combined with medication treatment, electroacupuncture combined with medication treatment and simple medication treatment in effects on rheumatoid arthritis pati...Objective： To compare differences of acupotomy loosing combined with medication treatment, electroacupuncture combined with medication treatment and simple medication treatment in effects on rheumatoid arthritis patients with elbow joint stiffness and investigate the anti-inflammatory mechanism of the acupotomy loosing.Methods： A total of 60 cases of rheumatoid arthritis（RA） patients with elbow joint stiffness were randomly assigned into the group receiving acupotomy loosing（group A）, group with electroacupuncture（group B） and the one undergoing medication treatment（group C） with 20 cases for each group. Based on the medication treatment, all patients underwent continuous oral administration with Methotrexate（MTX）, Leflunomide（LEF） and Bitongding capsules for 3 weeks. And no other treatments were given to group C. In addition to medications treatment, the electroacupuncture was performed in group B. The acupoints of Tianzhu（天柱 BL 10）,DAzhui（大椎 GV 14）; Fengchi（风池 GB 20）,Quchi（曲池 LI 11）,Quze（曲泽 PC 3）,Chize（尺泽 LU 5）, Shousanli（手三里 LI 10）, Xiaohai（小海 SI 8）, Shaohai（少海 HT 3）, Tianjing（天井 TE 10）,Qinglengyuan（清冷渊 TE 11） and Hegu（合谷 LI 4） in the affected side were selected. A pair of electrodes were connected to LI 11 and LI 10, and another pair of electrodes were connected to PC 3 and LU 5, and the continuous wave with frequency of 2 Hz was designed, the needle retention for 30 min was performed, and the acupuncture was performed for 6 times per week with 3 weeks for one course,and there was one course totally. Besides the medication treatment, group A underwent the acupotomy loosing therapy. The tender point in lateral elbow joint, etc. were taken as the treatment point, and 6-8 treatment points being taken for each time, and longitudinal dredging and transverse exfoliation were conducted for 2-3 times with the acupotomy. The treatment was given for one time per week with three weeks for one course, and there was one course totally. The changes of maximum angle of active extension position, maximum angle of active flexion position and range of motion were observed, and levels of IL-6, IL-10 and TNF-a in affected synovial fluid of elbow joint were tested before the treatment and 2 weeks after the treatment in the groups.Results： ① Compared with those before treatment, the maximum angle of active extension position were smaller, the maximum angle of active flexion position were larger and the range of motion were wider of the affected elbow joints of the patients in the 3 groups on the 2 weeks after the treatment.There were statistical significances for the differences（All P 0.05）. Two weeks after the treatment,as compared with those in group C, the affected elbow joint for patients in the group B and group A was smaller in maximum angle of active extension position, larger in maximum angle of active flexion position and wider in range of motion and there were statistical significances for the differences（All P 0.05）. Comparing with those in the group B, the affected elbow joint for patients in the group A was smaller in maximum angle of active extension position, larger in maximum angle of active flexion position and wider in range of motion and there were the statistical significances for the differences（All P 0.05）. ② Compared with those before treatment, the levels of TNF-a and IL-6 were lower and the level of IL-10 was higher of the 3 groups on the 2 weeks after the treatment. There were the statistical significances for the differences（All P 0.05）. For 2 weeks after the treatment, compared with those in the group C, group B and group A were lower in levels of TNF-α and IL-6 and higher in level of IL-10 and there were the statistical significances for the differences（All P 0.05）. As compared with those in the group B, group A was lower in levels of TNF-a and IL-6 and higher in level of IL-10 and there were the statistical significances for the differences（All P 0.05）.Conclusions： The combination of acupotomy loosing can improve the maximum angle of active extension position, maximum angle of active flexion position and range of motion in affected elbow joint for RA patients with elbow joint stiffness, whose efficacy was superior to single basic treatment and electroacupuncture combined with basic treatment. Meanwhile, the levels of proinflammatory cytokines,such as TNF-α and IL-6 can be decreased, and the level of anti-inflammatory cytokines, such as IL-10 can be increased, playing a role in regulating the imbalance between proinflammatory cytokines and anti-inflammatory cytokines in RA patients, which may be one of mechanisms regarding treating RA and improving the range of motion for stiff joints.展开更多
构建含人白介素10受体α(IL-10RA)基因的真核表达质粒p ENTER-IL-10RA-His,并在HEK293中进行真核表达,并用免疫共沉淀检测JAK1与IL10RA在细胞内的相互作用。在He La细胞中提取人总RNA,通过RT-PCR获得人IL10RA的基因全长,并将其克隆至真...构建含人白介素10受体α(IL-10RA)基因的真核表达质粒p ENTER-IL-10RA-His,并在HEK293中进行真核表达,并用免疫共沉淀检测JAK1与IL10RA在细胞内的相互作用。在He La细胞中提取人总RNA,通过RT-PCR获得人IL10RA的基因全长,并将其克隆至真核表达载体p ENTER-His中。经PCR扩增、双酶切、测序鉴定后,将重组质粒p ENTER-IL-10RA-His转染至HEK293细胞中。免疫印迹法检测IL-10RA蛋白在HEK293细胞中的表达。结果显示,经PCR扩增和双酶切,测序鉴定质粒克隆正确。免疫印迹可见63 k D的目的蛋白。共同转染JAK1和IL-10RA的质粒,免疫印迹可见分别为133 k D和63 k D的目的条带,免疫共沉淀鉴定了JAK1和IL-10RA的相互作用。IL-10RA基因成功构建在p ENTER-His中,并在HEK293细胞中成功表达,并成功共转染JAK1和IL-10RA质粒,免疫共沉淀检测两者的相互作用。这为JAK1和IL-10RA相互作用的机制研究奠定基础。展开更多
基金Supported by 2014 Wuhan clinical medical research project:WZ14C01~~
文摘Objective： To compare differences of acupotomy loosing combined with medication treatment, electroacupuncture combined with medication treatment and simple medication treatment in effects on rheumatoid arthritis patients with elbow joint stiffness and investigate the anti-inflammatory mechanism of the acupotomy loosing.Methods： A total of 60 cases of rheumatoid arthritis（RA） patients with elbow joint stiffness were randomly assigned into the group receiving acupotomy loosing（group A）, group with electroacupuncture（group B） and the one undergoing medication treatment（group C） with 20 cases for each group. Based on the medication treatment, all patients underwent continuous oral administration with Methotrexate（MTX）, Leflunomide（LEF） and Bitongding capsules for 3 weeks. And no other treatments were given to group C. In addition to medications treatment, the electroacupuncture was performed in group B. The acupoints of Tianzhu（天柱 BL 10）,DAzhui（大椎 GV 14）; Fengchi（风池 GB 20）,Quchi（曲池 LI 11）,Quze（曲泽 PC 3）,Chize（尺泽 LU 5）, Shousanli（手三里 LI 10）, Xiaohai（小海 SI 8）, Shaohai（少海 HT 3）, Tianjing（天井 TE 10）,Qinglengyuan（清冷渊 TE 11） and Hegu（合谷 LI 4） in the affected side were selected. A pair of electrodes were connected to LI 11 and LI 10, and another pair of electrodes were connected to PC 3 and LU 5, and the continuous wave with frequency of 2 Hz was designed, the needle retention for 30 min was performed, and the acupuncture was performed for 6 times per week with 3 weeks for one course,and there was one course totally. Besides the medication treatment, group A underwent the acupotomy loosing therapy. The tender point in lateral elbow joint, etc. were taken as the treatment point, and 6-8 treatment points being taken for each time, and longitudinal dredging and transverse exfoliation were conducted for 2-3 times with the acupotomy. The treatment was given for one time per week with three weeks for one course, and there was one course totally. The changes of maximum angle of active extension position, maximum angle of active flexion position and range of motion were observed, and levels of IL-6, IL-10 and TNF-a in affected synovial fluid of elbow joint were tested before the treatment and 2 weeks after the treatment in the groups.Results： ① Compared with those before treatment, the maximum angle of active extension position were smaller, the maximum angle of active flexion position were larger and the range of motion were wider of the affected elbow joints of the patients in the 3 groups on the 2 weeks after the treatment.There were statistical significances for the differences（All P 0.05）. Two weeks after the treatment,as compared with those in group C, the affected elbow joint for patients in the group B and group A was smaller in maximum angle of active extension position, larger in maximum angle of active flexion position and wider in range of motion and there were statistical significances for the differences（All P 0.05）. Comparing with those in the group B, the affected elbow joint for patients in the group A was smaller in maximum angle of active extension position, larger in maximum angle of active flexion position and wider in range of motion and there were the statistical significances for the differences（All P 0.05）. ② Compared with those before treatment, the levels of TNF-a and IL-6 were lower and the level of IL-10 was higher of the 3 groups on the 2 weeks after the treatment. There were the statistical significances for the differences（All P 0.05）. For 2 weeks after the treatment, compared with those in the group C, group B and group A were lower in levels of TNF-α and IL-6 and higher in level of IL-10 and there were the statistical significances for the differences（All P 0.05）. As compared with those in the group B, group A was lower in levels of TNF-a and IL-6 and higher in level of IL-10 and there were the statistical significances for the differences（All P 0.05）.Conclusions： The combination of acupotomy loosing can improve the maximum angle of active extension position, maximum angle of active flexion position and range of motion in affected elbow joint for RA patients with elbow joint stiffness, whose efficacy was superior to single basic treatment and electroacupuncture combined with basic treatment. Meanwhile, the levels of proinflammatory cytokines,such as TNF-α and IL-6 can be decreased, and the level of anti-inflammatory cytokines, such as IL-10 can be increased, playing a role in regulating the imbalance between proinflammatory cytokines and anti-inflammatory cytokines in RA patients, which may be one of mechanisms regarding treating RA and improving the range of motion for stiff joints.
文摘构建含人白介素10受体α(IL-10RA)基因的真核表达质粒p ENTER-IL-10RA-His,并在HEK293中进行真核表达,并用免疫共沉淀检测JAK1与IL10RA在细胞内的相互作用。在He La细胞中提取人总RNA,通过RT-PCR获得人IL10RA的基因全长,并将其克隆至真核表达载体p ENTER-His中。经PCR扩增、双酶切、测序鉴定后,将重组质粒p ENTER-IL-10RA-His转染至HEK293细胞中。免疫印迹法检测IL-10RA蛋白在HEK293细胞中的表达。结果显示,经PCR扩增和双酶切,测序鉴定质粒克隆正确。免疫印迹可见63 k D的目的蛋白。共同转染JAK1和IL-10RA的质粒,免疫印迹可见分别为133 k D和63 k D的目的条带,免疫共沉淀鉴定了JAK1和IL-10RA的相互作用。IL-10RA基因成功构建在p ENTER-His中,并在HEK293细胞中成功表达,并成功共转染JAK1和IL-10RA质粒,免疫共沉淀检测两者的相互作用。这为JAK1和IL-10RA相互作用的机制研究奠定基础。
