[Effect of electroacupuncture serum on expression of myogenic differentiation antigen and autophagy-related protein Beclin 1 in cultured muscle satellite cells under starvation].

OBJECTIVE: To explore the effect of electroacupuncture (EA) serum on expression of myogenic differentiation antigen (Myod) and autophagy-related protein Beclin 1 in cultured muscle satellite cells of rats under starvation conditions. METHODS: The primary multifidus muscle satellite cells of one male SD rat were isolated and cultured to obtain the 3rd generation of cells. The EA serum was got from the rat received EA stimulation of bilateral "Weizhong" (BL40, 2 Hz/10 Hz, 1 mA, duration of 20 min, once daily for 7 days). The cell suspension (2×104/well) of the 3rd generation of cultured cells was transferred to each well of a 96-well plate in medium containing 10% fetal bovine serum (FBS). Twelve duplicate wells were set up for the blank control serum (without FBS), 10% FBS, 10% EA serum, 20% EA serum and 30% EA serum groups and incubated for 12 h and 24 h, respectively. Each well was supplemented with 10 µL CCK-8 reagent to be incubated for 1 h again for observing the state of cell proliferation. After culturing the primary muscle satellite cells in serum-free medium for 12 h, the cells were randomly divided into serum-free group, 10% fetal bovine serum group and optimal concentration electroacupuncture serum group, and serum of corresponding concentration was added respectively. The expression levels of Beclin 1 and cell-proliferation-related protein Myod were detected by Western blot. RESULTS: CCK-8 assay displayed that the proliferation levels were significantly higher at 12 h and 24 h after serum intervention in the 10% FBS, 10% EA serum, 20% EA serum and 30% EA serum groups than that in the blank control serum group (P<0.01), and at 24 h in the 3 EA serum groups than in the 10% FBS group (P<0.01), but without significant difference among the three EA serum groups (P>0.05). As a result, 10% EA serum was selected as the optimal concentration for Western blot tests. No significant difference was found in the expression levels of Myod and Beclin 1 proteins among the serum-free, 10% FBS and 10% EA serum groups before intervention (P>0.05), and there was a marked up-regulation of Myod expression and an obvious down-regulation of Beclin 1 expression at 12 h in both the 10% EA serum and 10% FBS groups in comparison with their own pre-intervention (P<0.05). There were a marked up-re-gulation of Myod expression at both 12 h and 24 h and Beclin 1 expression at 24 h in both the 10% EA serum group and 10% FBS group than that in the serum-free group (P<0.05), and an obvious down-regulation of Beclin 1 expression at 12 h in both 10% FBS and 10% EA serum groups than that in the serum-free group (P<0.05, P<0.01). After 24 h's serum intervention, there was an increase of Myod expression and a reduction of Beclin 1 expression in both 10% FBS and 10% EA serum groups compared with those after the 12 h intervention (P<0.05). No significant differences were found between the 10% FBS and 10% EA serum groups in the expression levels of Myod and Beclin 1 proteins (P>0.05). CONCLUSION: EA serum can promote proliferation of cultured muscle satellite cells under starvation conditions, which is related to its functions in regulating expression of Beclin 1 and cell-proliferation-related protein Myod.

[Clinical trials of treatment of post-stroke foot drop by fire-needling plus stuck-needle-stretching].

OBJECTIVE: To compare the therapeutic effect of fire-needling plus stuck-needle-stretching and conventional acupuncture for post-stroke foot drop, so as to provide a reference foundation for optimizing clinical treatment regimen. METHODS: A total of 60 patients with post-stroke foot drop were equally randomized into a treatment group and a control group. In the treatment group, fire-needling plus stuck-needle-stretching was applied to acupoint pairs Jiexi (ST41)-Taichong (LR3), Xiajuxu (ST39)-ST41, Fenglong (ST40)-ST39, shangjuxu (ST37)-ST40, Zusanli (ST36)-ST37, Dubi (ST35)-ST36, Xuanzhong (GB39)-Qiuxu (GB40) on the affected side, and Chengshan (BL57) and Ashi points were swiftly pricked with red-hot filiform needle without needle retaining. In the conventional acupuncture group, the same acupoints were needled with filiform needles, and the treatment for both groups was given once a day, 6 times a week, for consecutive 4 weeks. The spasm severity of posterior triceps of the lower leg was evaluated using modified Ashworth scale, the tibial anterior muscle strength was measured using Lovett' and Martin's methods, the activities of daily living were assessed using modified Barthel's index, the walking ability was evaluated using Holden walking rating scale and the lower limb motor function assessed using Fugl-Meyer assessment scale, and the severity of foot drop was assessed in reference to Garceau criteria. RESULTS: After the treatment, the score of the modified Ashworth scale was significantly reduced (P<0.01), and those of Lovett muscle strength scale, Barthel index, Holden walking rating scale and Fugl-Meyer lower limb motor function scale were considerably increased in both groups versus their own pre-treatment (P<0.01). The therapeutic effect of the treatment group was significantly superior to that of the control group in reducing Ashworth scale score and in increasing the scores of Lovett muscle strength test, Barthel index, Holden walking function scale and Fugl-Meyer lower limb motor function scale (P<0.01). Of the two 30 cases in the control and treatment groups, 10(33.33%)and 14(46.67%) experienced a remarkable improvement, 10(33.33%)and 12(40.00%)were improved, 7(23.33%)and 3(10.00%)had a mild progress, 3(10.00%) and 1(3.33%) failed, with the excellent plus good effective rates being 66.67% and 86.67%, respectively, but without significant difference between the two groups (P>0.05). CONCLUSION: Both fire-needling plus stuck-needle-stretching and conventional acupuncture are effective in the treatment of post-stroke foot drop, but the former is relatively better.