[Application characteristics and modern research progress of "bone-approaching" acupuncture].

The paper explores the evolution of "bone-approaching" acupuncture, its effect target and mechanism. The concrete operation procedure of "bone-approaching" method is recorded originally in Huangdi Neijing (Inner Canon of Yellow Emperor) as short needling and Shu needling (referring to the category of the five needling technique). The periosteum is the most effective stimulation target of "bone-approaching" acupuncture for analgesia, regaining consciousness and regulating spirit. The "bone-approaching" acupuncture is not only prominently effective on bone bi syndrome, but also has the unique effect on painful, encephalogenic and emotional diseases. The paper summarizes and improves "bone-approaching" acupuncture, i.e. "touching bone surface" with needle tip by slow insertion, "touching bone surface" without pain by swift insertion and "touching bone" with needle body by oblique insertion. It contributes to the inheritance, development and supplementation to the bone needling techniques in Huangdi Neijing and is significant for broadening the clinical application range of acupuncture.

[Discussion on clinical application of "touching-bone" acupuncture technique].

The value of the "touching-bone" acupuncture technique in clinical application was explained through the investigation on the origin of the theory of the "touching-bone" acupuncture technique, the analysis on the characteristics of acupoint selection, the introduction of clinical characteristics and the discussion on the mechanism of acupuncture in treatment. The "touching-bone" acupuncture technique refers to deep needling method, originated from the short needling and shu needling of the ancient needling methodslisted in the Internal Classic. The target points are the reaction sites on meridian near to bone and the attachments of soft tissues on bone. During the needle insertion, the needle tip is thrust deeply to the bone or the needle body is closely attached to the bone so as to stimulate periosteum specifically. This needling technique contributes to the satisfactory effect on spasmodic, deep-located and intractable pain disorder, motor system diseases, mental diseases and cerebral diseases, etc. Hence, this acupuncture technique deserves to be promoted in clinical application and explored in research.

[Effects of electroacupuncture at "Weizhong" (BL 40) on morphology and expression of CK and IL-17 in rats with bupivacaine-induced multifidus muscle injury].

OBJECTIVE: To observe effects of electroacupuncture (EA) at "Weizhong" (BL 40) on morphology and expression of creatine kinase (CK) and interleukin-17 (IL-17) in rats with bupivacaine-induced multifidus muscle injury. METHODS: A total of 32 male SD rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 8 rats in each one. The rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine to establish the model of multifidus muscle injury; the rats in the control group were injected with 0.9% sodium chloride solution. The rats in the Weizhong group and Shenshu group were treated with EA (2 Hz/10 Hz in frequency, 1~2 mA in intensity) at "Weizhong" (BL 40) and "Shenshu" (BL 23), 20 min per treatment. No treatment was given in the control group and model group. After 14-day treatment of EA, the inflammatory cell count, scar tissues area and muscle fiber cross sectional area of multifidus muscle were observed with HE and Masson staining method. The activity of CK and serum content of IL-17 were test with enzyme-linked immunosorbent assay (ELISA) method; the expression of IL-17 in multifidus muscle was measured with immunohistochcmical method. RESULTS: After intervention, the inflammatory cell count and scar tissues area in the model group, Weizhong group and Shenshu group were higher than those in the control group (all P<0.01), but the muscle fiber cross sectional area was significantly reduced (all P<0.01); the inflammatory cell count and scar tissues area in the Weizhong group and Shenshu group were lower than those in the model group (all P<0.01), and the muscle fiber cross sectional area was significantly increased (P<0.01, P<0.05). After intervention, the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the model group, Weizhong group and Shenshu group were higher than those in the control group (all P<0.01); the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the Weizhong group and Shenshu group were lower than those in the model group (P<0.01, P<0.05); compared with the Shenshu group, the down-regulation of IL-17 was more obvisous in the Weizhong group (P<0.01). CONCLUSION: EA at "Weizhong" (BL 40) can down-regulate the overexpression of serum CK and IL-17, alleviate inflammation reaction and improve the repair of multifidus muscle.

[Effects of electroacupuncture at "Weizhong" (BL 40) on regeneration and morphology in rats with bupivacaine-induced multifidus muscle injury].

OBJECTIVE: To observe the intervention effect of electroacupuncture (EA) at "Weizhong" (BL 40) on rats with bupivacaine-induced multifidus muscle injury, so as to explore the action mechanism. METHODS: A total of 72 rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 18 rats in each group. Each group was again randomly divided into a 4-day subgroup, a 7-day subgroup and a 14-day subgroup, 6 rats in each subgroup. Rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine (BPVC) to establish the model of multifidus muscle injury. Rats in the Weizhong group and Shenshu group were treated with EA at "Weizhong" (BL 40) and "Shenshu" (BL 23), 20 min per treatment, once a day. Each subgroup was treated for 4 days, 7 days and 14 days respectively. Rats in the control group and model group were treated with immobilization. The morphology and cross sectional area (CSA) changes of multifidus with HE and Masson staining at different time points were observed; the expression of insulin like growth factor 1 (IGF-1) and myogenic differentiation antigen (MyoD) was measured by immunohistochemical method. RESULTS: After the modeling, there were significant morphology changes of multifidus at different time points, which was not fully recovered after 14 days. The morphological observation in the Weizhong group and Shenshu group was superior to that in the model group. At 7th day, the CSA in the Weizhong group was higher than that in the model group (P < 0.05); at 14th day, the CSA in the Weizhong group and Shenshu group was higher than that in the model group (P < 0.01, P < 0.05). At 4th day and 7th day, the expression of IGF-1 in the model group was higher than that in the control group (both P < 0.01); at 4th day, that in the Weizhong group was higher than that in the model group (P < 0.01), and that in the Weizhong group was higher than that in the Shenshu group (P < 0.05), and that in the Shenshu group was as higher than that in the model group (P < 0.05); at 14th day, that in the Shenshu group was higher than that in the model and Weizhong group (P < 0.01). At 4th day, the expression of MyoD in the Weizhong group and Shenshu group was higher than that in the model group (P < 0.01), which was more significant in the Weizhong group (P < 0.01). CONCLUSION: Electroacupuncture at "Weizhong" (BL 40) and "Shenshu" (BL 23) can both promote the regeneration of multifidus muscle injury. EA at "Weizhong" (BL 40) has a better effect at early phase, which may be related to the up-regulation of IGF-1 and MyoD and the completion of the proliferation of myoblast in advance.

[Effect of Electroacupuncture Serum on Proliferation of Cultured Multifidus Muscle Satellite Cells and Expression of Pax-7, MyoD and p-Akt].

OBJECTIVE: To observe the effect of electroacupuncture (EA) serum on proliferation of multifidus muscle sa-tellite cells (SCs) and expression of paired box transcription factor Pax-7, MyoD and protein kinase B (PKB or Akt) proteins of SCs, so as to explore its underlying mechanism in promoting repair of multifidus muscles. METHODS: Thirty-two SD rats were randomly assigned to control, model, EA-Weizhong (BL 40) and EA-Shenshu (BL 23) groups. The multifidus muscle injury (MFMI) model was established by injection of 0.5% bupivacaine hydrochloride (400 µL) into the bilateral L4-L5 paravertebral muscles (4 points, 100 µL for each point). EA stimulation was separately applied to bilateral BL 40 and BL 23 for 20 min, once daily, 4 days altogether. Blood samples of the abdominal artery of rats in the above mentioned 4 groups were separately collected for extracting serum, followed by deactivation and filtration, and then were respectively applied to the Dulbecco's Modified Eagle Media (DMEM) culturing each multifidus muscle SCs of the normal serum, model serum, EA-BL 40 serum and EA-BL 40 serum+LY 294002 (an inhibitor of phosphotidylinsitol-3-kinase, PI 3 K), EA-BL 23 serum and EA-BL 23 serum+LY 294002 groups for ana-lyzing the impact of EA serum on the proliferation state of SCs by Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU) methods, respectively. The expression of Pax-7, MyoD and phosphorylated (p)-Akt proteins of the cultured SCs was detected for characterization of SCs by Western blot. RESULTS: Compared with the normal serum group, the proliferation levels (detected by both CCK-8- and EdU) and the expression levels of MyoD and p-Akt proteins of SCs in the model serum group were significantly increased (P<0.05, P<0.01), while in comparison with the model serum group, the proliferation and expression levels of MyoD and p-Akt proteins of SCs were further significantly increased in both EA-BL 23 and EA-BL 40 serum groups (P<0.01, P<0.05), but not in the EA-BL 40 serum+LY 294002 and EA-BL 23 serum+LY 294002 groups (P>0.05), suggesting an involvement of PI 3 K in the proliferation of SCs. No marked differences were found in the proliferation levels between the EA-BL 23 and EA-BL 40 serum groups and in the expression levels of Pax-7 proteins among the 6 serum groups (P>0.05). CONCLUSIONS: Both EA-BL 40 and EA-BL 23 serum can promote proliferation of multifidus muscle SCs, which may contribute to the effect of EA intervention in promoting repair of the injured muscle, partially by way of Akt/PI 3 K signaling.