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Objective To observe the effects of massage on inflammation,oxidative stress and autophagy during the repair of acute contusion of skeletal muscles so as to explore its biological mechanisms.Methods Forty-two adult Sprague-Dawley rats were randomly divided into a control group (n =6),a model group (n =18),and a treatment group (n =18).Acute contusion of the gastrocnemius muscles of the rats in the model and treatment groups was inflicted using a home-made impactor.Beginning forty-eight hours later,15 minutes of massage was administered daily for two weeks.After one,7 and 14 days of the massage treatment,the injured gastrocnemius was resected from 6 rats of both the model and treatment groups.Morphological changes were observed using haematoxylin and eosin (HE) staining.The serum content of tumor necrosis factor alpha (TNF-α),interleukin1β (IL-1 β),C reactive protein (CRP) and prostaglandin E2 (PGE2) were detected using enzyme-linked immunosorbent assay (ELISA).The serum content of superoxide (SOD) and malondialdehyde (MDA) were detected using spectrophotometry.The expression of microtubule-associated protein 1 light chain 3 (LC3),Bcl-2 homeodomain protein Beclin1 and ubiquitin binding protein P62 were detected using Western blotting.Results The HE staining showed more significant collapse and swelling of cells in the model group than in the control group at each time point.New muscle cells were observed at days 7 and 14 in the model group.At each time point,significantly better recovery was observed in the treatment group compared to the model group,with more new muscle cells and better cell morphology.According to the ELISA results,a significant increase in serum pro-inflammatory factors occurred in the model group compared to the control group and compared to the treatment group after one day and 7 days of treatment.The average serum content of SOD and MDA in the model group was significantly higher than in the control group,while the average serum content of SOD in the treatment group was significantly higher than in the model group and that of MDA was significantly lower.Western blotting showed a significant decrease in LC3 (Ⅱ/Ⅰ) and Beclin1,as well as a significant increase in P62 in the model group at each time point compared with the treatment group and the controls.Conclusion Inflammation and oxidative stress increase significantly in a skeletal muscle after injury,but autophagy decreases significantly.Massage can effectively reduce the inflammatory response and oxidative stress and promote autophagy,which leads to quicker repair of skeletal muscles.
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@# Irrespective of physiological or pathological skeletal muscle atrophy, the endocrine and motor functions of skeletal muscle are impaired. The mechanism of acupuncture to prevent and treat skeletal muscle atrophy is not only related to classical protein synthesis and decomposition, but also involves apoptosis, autophagy, muscle satellite cell proliferation and differentiation, muscle fiber type conversion, neuromuscular junction conduction, and cell energy metabolism conversion.
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@#Objective To explore the effects and mechanism of electroacupuncture (EA) on denervation-induced atrophy in rats. Methods A total of 18 male Sprague-Dawley rats were divided into sham group (n=6), model group (n=6) and EA group (n=6). The latter two groups were clamped right sciatic nerve to establish atrophy model of skeletal muscle. On the second day after modeling, EA group accepted electroacupuncture on right Zusanli (ST36) and Huantiao (GB30) for two weeks. Their gastrocnemius muscles were obtained after intervention, and the wet weight ratio of the gastrocnemius muscles was calculated. The cross-sectional area (CSA) and diameter of muscle fibers were measured after HE staining. The protein expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), 70-KD ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6k (p-p70S6k) was tested with Western blotting. The gene expression of mTOR and p70S6K was detected with real-time quantitative polymerase chain reaction. Results Compared with the sham group, the wet weight ratio of the gastrocnemius muscle, CSA and diameter of the muscle fibers decreased in the model group and EA group (P<0.001), which were more in EA group than in the model group (P<0.01); the protein expression of mTOR, p-mTOR, p70S6K and p-p70S6K increased in the model group (P<0.01), and increased more in EA group (P<0.05); the gene expression of mTOR and p70S6K increased in the model group (P<0.05) , and increased more in EA group (P<0.05).Conclusion Electroacupuncture delays the atrophy of denervated skeletal muscles, which may relate to activation of mTOR/p70S6K signal pathway to impact synthesis of skeletal muscle proteins.
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<p><b>OBJECTIVE</b>To observe the effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor (HGF) in rats with skeletal muscle contusion, and to explore the mechanism of acupuncture at opposite acupoints on skeletal muscle contusion.</p><p><b>METHODS</b>Fifty-four Sprague Dawley (SD) rats were randomly divided into a blank group (6 rats), a model group (24 rats) and an opposing needling group (24 rats). The model group and opposing needling group were further divided into 1-day subgroup, 3-day subgroup, 5-day subgroup and 7-day subgroup, 6 rats in each one. No intervention was given in the blank group, while the model of skeletal muscle contusion was established in the model group and opposing needling group by self-made contusion device. 24 hours after contusion, electroacupuncture (EA) was applied at "Zusanli" (ST 36) and the corresponding points ofpoints at health side for 15 min, once a day. The subgroups of opposing needling group were treated for 1 day, 3 days, 5 days and 7 days, respectively. No treatment was given in the model group. Samples were collected in the subgroups 1 day, 3 days, 5 days and 7 days after treatment. The morphological change of injured gastrocnemius muscle was observed by using microscope after HE staining. The positive cell rate of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. The expression levels of HGF protein and PCNA protein were observed by Western blot.</p><p><b>RESULTS</b>① The results of HE staining showed that, 1 day after contusion, the inflammatory cells of gastrocnemius muscle in the opposing needling group were less than those in the model group; 3 days and 5 days after contusion, myoblasts and myotubes in the opposing needling group were more than those in the model group; 7 days after contusion, the neonatal muscle cells in the opposing needling group were more than those in the model group. ② The results of immunohistochemistry showed that, 1 day, 3 days and 5 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly higher than that in the model group (all<0.001); 7 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly less than that in the model group (<0.001). ③ The results of Western blot showed that, 1 day, 3 days and 5 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly higher than that in the model group (all<0.05); 7 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly lower than that in the model group (all<0.05).</p><p><b>CONCLUSION</b>Acupuncture at opposite acupoints could regulate the expression of HGF and promote the activation, proliferation, migration and differentiation of muscle satellite cells in rats with skeletal muscle contusion, which could speed up the process of skeletal muscle injury repair.</p>
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Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.
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Traditional Chinese Medicine as a rare heritage from our progenitor is becoming more and more important in the world. A lot of foreign students have come to China for learning TCM , and the number of foreign students has increased with time. A lot of issues occur during the teaching process for the foreign students. The problems including the source of students' quality, language, teaching material and clinic practice during the teaching process for the foreign students were ana-lyzed in the article, and the resolutions according to the problems were put forward to attract more and more foreign students to learn TCM and promote TCM in the world.
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Objective To explore the curative effect of acupuncture and ultrasonic therapy on treatment of the menopausal osteo -porosis patients who suffered from pain in waist and back .Methods 120 cases menopausal osteoporosis patients from outpatients were divided into 2 groups randomly .Acupuncture and ultrasonic therapy were used in the treatment group ,soybean isoflavone was only supplied in the control group .Then pain intension and bone density in lumbar vertebrae were observed .Results Compared with control group ,the pain in treatment group relieved more effective (P< 0 .05) .The bone density in treatment group was higher than that in control group (P< 0 .05) .The efficient of treatment group is higher than control group (P < 0 .05) .Conclusion The method of acupuncture and ultrasonic therapy can improve the bone density and relieve pain .It can treat the menopausal osteoporo-sis effectively .
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Objective To investigate correlation between demyelination and caspase-12 expression alteration after compressed spinal cord injury (CSCI) so as to discuss mechanism of demyelinating lesion after CSCI.Methods Seventy-five adult SD rats were randomly divided into five groups,ie,normal group,control group,compression 1 d,3 d and 7 d groups,with 15 rats per group.Models of spinal cord compression were established with a self-made device.Ultrastructure of the demyelinated nerve fibers was observed by electronic microscope and oligodendrocyte apoptosis was detected by TUNEL staining and double labeling immunofluorescence.Immunoblotting was used to defect caspase-12 that was related to cell apoptosis.Results Demyelination of nerve fiber occurred after CSCI and was aggravated with time.Apoptosis of oligodendrocytes was found after CSCI,and showed significant difference between compression 7 d group and normal group (P ＜ 0.05).Caspase-12 was also upregulated with extension of compression time.Conclusion Caspase-12 mediating oligodendrocyte apoptosis is one of the mechanisms of nerve fiber demyelination after CSCI.
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Objective To observe the effects of electro-acupuncture on the expression and inhibition of DNA binding protein 2 (Id2) and myelin basic protein (MBP),and to explore the mechanism of remyelinization after compressive spinal cord injury (CSCI) in rats.Methods Fifty-four SD rats were randomly divided into a control group and a treatment group,and each was further subdivided into 3 time point subgroups:3,7 and 14 days.There were 9 rats in each subgroup.The CSCI models were made with a self-designed method.The acupuncture points Jiaji (EX-B2),bilateral Zusanli (ST36) and Taixi (KI3) were selected for treatment.Electro-acupuncture (continuous wave,2 Hz,1.5 V)was applied to the bilateral Zusanli (ST36) and Taixi (KI3) points.The control group received the injury but no treatment.The changes in the ultrastucture of the nerve fibers＇ white matter were de-termined by transmission electron microscopy (TEM).The alterations in the expression of MBP and Id2 were observed by double labeled immunofluorescence and Western blotting on the 3rd,7th and 14th day after the injury.Results TEM showed that the myelin sheaths in the control group had degenerated,swollen,and even broken down after CSCI.Changes to the myelin sheaths in the treatment group were milder than those in the control group.The immunofluorescence results showed the amount of Id2-immunoreactive oligodendrocytes in the control group to be (20 ±2) on the 3rd day after CSCI,becoming (16 ± 1) on the 14th day.The differences among the 3 control subgroups were not statistically significant.The amount of Id2-immunoreactive oligodendrocytes in the treatment group was (13 ± 1) on the 3rd day,reaching a minimum the 14th day.The differences among the 3 treatment groups were statistically significant.The differences compared with the control group at the same time points were also statistically significant.Western blotting showed that the expression of Id2 in the contrast and treatment groups was (1.12 ±0.12) and (0.67 ±0.01) respectively on the 3rd day after CSCI,and both decreased with time.The expression of Id2 in both groups reached their minima ((0.86 ±0.02) and (0.25 ±0.01) respectively) on the 14th day.The difference between the treatment groups and the contrast group was statistically significant at each time point.The expression of MBP in the contrast and treatment groups at day 3 was (0.44 ± 0.02) and (0.67 ± 0.04) respectively,and these increased with time.The expression of MBP in both groups peaked at the 14th day (at (0.95 ± 0.04) and (1.74 ± 0.09) respectively).These differences were again statistically significant.Conclusion Electro-acupuncture can regulate the expression of Id2 and MBP after CSCI.The down-regulation of Id2 which controls MBP negatively and the up-regulation of MBP may contribute to remyelination in the injured spinal cord.
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Objective To investigate the role of demyelination and the alteration of chondrotin sulfate proteoglycan (CSPG,NG2) expression after compression injury of the spinal cord (CSCI).Methods Seventy-five adult Sprague-Dawley rats were randomly divided into a normal group,a sham-operation group,a CSCI 1 day group,a CSCI 3 day group,and a CSCI 7 day group.There were 15 rats in each group.The injuries in the CSCI groups were inflicted using a technique devised in our laboratory.Basso-Beattie-Bresnahan (BBB) neurological function assessment was used to assess the rats' motor function,osmic acid staining and transmission electronic microscopy (TEM)were used to observe any pathological changes of myelinated nerve fibers in the white matter at 1,3 and 7 days after CSCI.The amount of myelinated nerve fibers in the posterior funiculus of the spinal cord and the ratio of myelin sheath thickness to axon diameter (the G-ratio) were calculated.Any alteration in NG2 expression was observed by Western blotting.Results The average neurological function assessment scores in the CSCI groups were (1.23 ±0.45),(0.65 ± 0.35) and (0.00 ± 0.00) respectively.Compared with the normal group (21.00 ± 0.00) and the sham operation group (21.00 ± 0.00),the differences were all statistically significant.The rats' motor function deteriorated gradually with time after the CSCI.Osmic acid staining showed that the white matter was intact in the normal and sham groups.After being compressed the myelinated nerve fibers became swollen,degenerated and broke down.The amount of myelinated nerve fibers in the normal group,the sham operation group and the three CSCI groups was (2771 ± 108),(2675 ± 199),(2403 ± 161),(1708 ± 70) and (8 10 ± 95) respectively.The amount of myelinated nerve fibers decreased in the CSCI groups and reached a minimum on the 7th day.The difference was statistically significant.The TEM quantity analysis showed that the G-ratios in the normal,sham operation,and CSCI 1 day,3 day and7 day groups were (18.10±0.4),(17.70±1.0),(6.69 ±0.8),(5.73 ±0.4) and (4.95 ±0.5) respectively.Compared with the normal and sham operation groups,the G-ratios in the 3 CSCI groups were lower and reached their minimum on the 7th day after injury.The difference was statistically significant.TEM observation showed that the axons and myelin sheaths were intact in the normal and sham groups.After CSCI the axons became swollen and cell organelles in the axoplasm degenerated and decreased.The layers of myelin sheath shrank,folded and even wrinkled,which had an onion-like appearance.The oligodendrocytes exhibited chromatin condensation.Macrophages showed infiltration.Western blotting showed that the expression of NG2 in the CSCI groups reached a maximum on the 1st day after injury and then decreased with time.The expression of NG2 in the CSCI groups was higher than in the normal and sham groups,and the difference was statistically significant.Conclusion Demyelination occurs after CSCI-the amount of myelinated nerve fibers decreases and neurological deficits increase with time.The expression of NG2 was associated with changes in the myelin sheaths after CSCI and contributed to recovery of the myelin sheath through proliferation and differentiation to oligodendrocytes and perhaps other kinds of cells.
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The TCM acupuncture education for foreign students is complicated and technical work.The writer discusses the modern teaching methods recently to aim directly at the problems in the traditional teaching methods in foreign students education,analyzes and compares the connotation and teaching mode of these teaching methods,and at last puts forward that we should choose correct teaching method according to the actuality to improve the teaching quality for foreign students in TCM acupuncture.