[Effect of acupuncture pretreatment of "Quchi"(LI11) and "Xuehai" (SP10) on mast cells and IL-33/ST2 in rats with urticaria].

OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment at "Quchi "(LI11) and "Xuehai "(SP10) on expression of interleukin (IL)-33, suppression of tumorigenicity 2 (ST2) and mast cell degranulation in sensitive area of skin tissue in rats with urticaria, so as to explore its mechanisms underlying prevention of urticaria. METHODS: A total of 32 male SD rats were randomly divided into blank control, model, EA preconditioning and medication groups, with 8 rats in each group. The urticaria model was established by topical injection of the prepared anti-ovalbumin serum (foreign serum, 0.1 mL/spot) along the bilateral sides of the spinal column on the back, followed by injection of mixture solution of ovalbumin, 0.5% evans blue and normal saline via the tail vein 48 h later. EA intervention (2 Hz/15 Hz, 1 mA) was applied to bilateral LI11 and SP10 for 20 min, once daily for 7 d before modeling.Back sensitization was started from the 5th day on. Rats of the medication group received gavage of loratadine, and those of the model group received gavage of the same volume of normal saline. The diameter of evans blue spots at the back skin tissue was measured; the histopathological changes of the blue spot tissues were observed by light microscope after H.E. staining. The state of degranulation of mast cells in the subcutaneous loose connective tissue was observed by using toluidine blue staining. Serum IgE and histamine contents were detected by ELISA, and the immunoactivity of IL-33 and ST2 in the skin and subcutaneous tissues of the sensitized spots (evans blue exudation spots) was observed by immunohistochemistry. RESULTS: Compared with the blank control group, the diameter of evans blue spot, degranulation rate of mast cells, serum IgE and histamine contents, and the immunoactivity of IL-33 and ST2 in the evans blue exudation spot tissues were significantly increased in the model group (P<0.01). In comparison with the model group, the increase of the above-mentioned indexes was reversed in both EA and medication groups (P<0.01，P<0.05). No significant differences were found between the EA and medication groups in down-regulating the levels of the 6 indexes. H.E. staining of the blue spot tissues of rats in the model group showed incomplete structure of the epidermal layer of the skin, unclear interface of tissues, incomplete keratinization, chaotic epidermal cells, disorderly arrangement of fibers in the dermis, and infiltration of inflammatory cells and edema, which was relatively milder in the EA and medication groups. CONCLUSION: EA preconditioning can prevent urticaria (reduce size and sensitive reactions) in rats, which may be associated with its functions in lowering the level of IgE through inhibiting IL-33 and ST2.

[Electroacupuncture preconditioning relieves cutaneous passive anaphylaxis by down-regulating IP3 mediated ROS/TRPM2 signaling pathway and inhibiting mast cell degranulation in rats with urticaria].

OBJECTIVE: To observe the effect of electroacupuncture (EA) preconditioning of "Quchi" (LI11) and "Xuehai" (SP10) on mast cell (MC) degranulation, and expressions of inositol triphosphate(IP3), reactive oxygen species (ROS), transient receptor potential (TRP) M2, calmodulin (CaM) in rats with urticaria, so as to reveal its molecular mechanism under-lying improving urticaria. METHODS: Thirty-two male SD rats were randomly divided into blank control, model, preconditioning of EA (Pre-EA) and medication groups (n=8 rats/group). The urticaria model was established by intradermal injection of dilute allogeneic antioalbumin serum at the spots of the bilateral symmetry of the spine on the back, and followed by tail venous injection of mixture solution of egg albumin diluent, plus 0.5% Evans blue and normal saline. Ten days before the end of modeling, rats of the pre-EA group received EA stimulation of LI11 and SP10 for 20 min, once a day for 10 consecutive days, and those of the medication group received gavage of loratadine tablets diluted solution (1 mg/kg) once a day for 10 days. The times of rat's scratching the sensitized skin were recorded, the diameter of the sensitized blue spots was measured and the degranulation rate of skin MCs was counted under microscope after toluidine blue staining. The expression levels of IP3, ROS, TRPM2 and CaM in the skin tissue were measured by immunohistochemistry and western blot, respectively. RESULTS: Compared with the blank control group, the scratching times, diameter of the sensitized blue spots, degranulation rate of MCs, and the expression levels of ion channel related proteins (IP3, ROS, TRPM2 and CaM) were significantly increased (P<0.01) in the model group. In comparison with the model group, the scratching times, diameter of sensitized blue spot, degranulation rate of MCs, and the expression levels of IP3, ROS, TRPM2 and CaM in both pre-EA and medication groups were significantly down-regulated (P<0.01, P<0.05). No significant differences were found between Pre-EA and medication groups in down-regulating the levels of the above-mentioned 7 indexes. CONCLUSION: EA-LI11 and SP10 preconditioning can reduce the cutaneous anaphylaxis in urticaria rats, which may be related to its effects in inhibiting the degranulation of MCs, and the expression of TRP channel related proteins.

[Current status, challenges and countermeasures of acupuncture data mining literature researches].

Data mining is an analytical method for revealing the implicit internal relations among the data elements, and is also widely used in the field of acupuncture and moxibustion of traditional Chinese medicine. However, there exist some significant deficits in the rationality of design and implementation, preciseness, repeatability, comprehensiveness, objectivity and depth of insights in some current acupuncture data mining researches. In the present paper, we summarized the literature on acupuncture data mining published in the past five years, and analyzed their common shortcomings in the design, implementation and reporting process, including too broad research scope, fuzzy and limited descriptions about the inclusion criteria, not definite retrieval scope and strategy, rarely seen original researches about the assessment of the report quality, lack of detailed descriptions about the literature screening and data processing procedure, incomplete narration about the research outcomes and their clinical significance, and lack of comprehensiveness and subjectiveness and available coping strategies in the discussion of the research papers, etc., in order to promote the improvement of literature methodology and quality of acupuncture data mining research, and then improve the reliability and clinical reference value of such research results.

[Influence of electroacupuncture of"Zusanli"(ST36)on mast cells/TRPV1 signaling pathway in visceral hypersensitivity rats with functional dyspepsia].

OBJECTIVE: To explore the interventional mechanism of electroacupuncture (EA) of "Zusanli"(ST36)based on the involvement of mast cells/ transient receptor potential vanilloid type1 (TRPV1) signaling pathway in relieving visceral hypersensitivity in functional dyspepsia (FD) rats. METHODS: Sixty SD rats (half male and half female, 10 days in age) were randomly divided into normal control, model, medication (ketotifen) and EA groups, with 15 rats in each group. The FD model was established by gavage of iodoacetamide combined with tail clamping (stress stimulation). Rats of the medication group received intraperitoneal injection of ketotifen (1 mg·kg-1·d-1) for 14 d, and those of the EA group received EA of ST36 for 20 min, once a day for 14 d. An air-balloon was inserted into the rat's stomach for recording changes of the intragastric pressure (mL/mm Hg) via a pressure transducer. The visceral hypersensitivity was assessed using abdominal withdrawal reflex (AWR) score and the number and degranulation of mast cells of gastric mucosa were observed using toluidine blue staining. The expression levels of TRPV1 and proteinase activated receptor 2 (PAR2) in the stomach were observed using immunofluorescence histochemistry and Western blot, separately, and the contents of SP and CGRP in the stomach detected using ELISA. RESULTS: When the intragastric pressure was at 50, 60 and 70 mm Hg, the gastric compliance was significantly decreased (P<0.01), and the levels of visceral sensitivity increased in the model group （P<0.01）。 TRPV1 immunofluorescence tensity, expression of PAR2 and TRPV1 proteins, and contents of SP and CGRP in the stomach were considerably up-regulated in the model group compared with the normal control group (P<0.01). In comparison with the model group, under intragastric pressure of 50，60 and 70 mm Hg, the gastric compliance was obviously increased, and the visceral hypersensitivity decreased in the EA group （P<0.01,P<0.05）. TRPV1 immunofluorescence intensity, expression levels of PAR2 and TRPV1 proteins, and the contents of SP and CGRP in the stomach were considerably down-regulated in both medication and EA groups compared with the model group (P<0.01, P<0.05). The therapeutic effect of EA was significantly superior to that of medication in up-regulating the gastric compliance （at 70 mm Hg）, and down-regulating the contents of SP and CGRP (P<0.05). No significant differences were found between the EA and medication groups in up-regulating gastric compliance at intragastric pressure of 50 and 60 mm Hg, and in down-regulating the visceral sensitivity, TRPV1 fluorescence intensity, and expression of PAR2 and TRPV1 proteins (P>0.05). Toluidine blue staining showed an apparent increase of mast cell number with obvious degranulation in the gastric mucosa of rats in the model group, which was milder in the EA and medication groups. CONCLUSION: EA of ST36 can suppress visceral hypersensitivity and increase the gastric compliance in FD rats, which may be related with its effects in inhibiting the activation of gastric mast cells, and down-regulating the expression of gastric PAR2 and TRPV1 proteins and SP and CGRP contents.

Effectiveness and safety of acupuncture for chronic obstructive pulmonary disease: A protocol for systematic review and meta-analysis.

BACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is the most common respiratory disease in clinic. Traditional Chinese medicine (TCM) lung rehabilitation has gradually been valued in the field of prevention and treatment of COPD. Acupuncture, as an important part of TCM lung rehabilitation, is carried out in clinical extensively and application. It has the advantage of improving the symptoms and quality of life of COPD cases, but the evidence-based proof is limited. METHODS: Eight databases will be systematically retrieved from their inceptions to December 2021. This study will include randomized controlled trials of acupuncture in the treatment of COPD with stable stage. The main outcome will include clinical effective rate, TCM symptom score, quality of life, dyspnea, exercise capacity, lung function, frequency of acute exacerbation, adverse events. Literature screening, quality evaluation and information extraction will be completed by two independent researchers. Revman 5.3 software will be used to perform meta-analysis. Assessment of multiple systematic reviews-2 tool will be used for evaluating the methodological quality of systematic review (SR), grading of recommendations assessment will for rating the quality of evidence, and consolidated standards of reporting trials and Standards for Reporting Interventions in Clinical Trials of Acupuncture tool will for assessing the reporting quality of randomized controlled trials. RESULTS: The findings of this SR will be presented at relevant conferences and submitted for peer-review publication. CONCLUSIONS: The findings of this SR will provide up-to-date summary proof for evaluating the effectiveness and safety of acupuncture for COPD.

[Effect of moxibustion on the expression of microglia and brain-derived neurotrophic factor protein in the spinal cord of rats with myofascial pain syndrome].

OBJECTIVE: To observe the effect of moxibustion at myofascial trigger points on microglia activation and the expression of brain-derived neurotrophic factor (BDNF) in the spinal cord of rats with myofascial pain syndrome (MPS), so as to explore the central mechanism of the analgesic effect of moxibustion. METHODS: Twenty-four male SD rats were randomly divi-ded into control, model and moxibustion groups (n=8 in each group). The MPS model was established by strinking on fascia musculares and eccentric exercise. Rats in the moxibustion group were treated with mild moxibustion at myofascial trigger point for 15 min, once daily for 7 days. The thermal withdrawal latency (TWL) of rats was measured with a hot stabbing instrument. The pathological changes of the rat medial femoris muscle were observed after H.E. staining. The expressions of microglia marker (OX-42) and BDNF in the spinal dorsal horn were detected by immunohistochemistry and Western blot, separately. RESULTS: After modeling, the TWL of both the model and the moxibustion groups were significantly down-regulated (P<0.01)，and were significantly decreased in contrast to that of the control group (P<0.01). After treatment and compared with the model group, the TWL of the moxibustion group was significantly increased (P<0.01) in the moxibustion group. Rat's muscle fibers of the control group were uniform in thickness and arranged tightly and regularly. While in the model group, some fractures and connective structure tissue renewal, irregular arrangement, and inflammatory cell infiltration were seen. The morphology of muscle fibers in the moxibustion group was close to normal, and the arrangement was neat and orderly, with a small amount of inflammatory cell infiltration. Compared with the control group, the expression of OX-42 and BDNF in the spinal dorsal horn of rats in the model group was increased(P<0.01). Following the treatment, and in comparison with the model group, the expression of OX-42 and BDNF of moxibustion group was down-regulated(P<0.05). CONCLUSION: Moxibustion can significantly improve the injury of the medial femoral muscle and the TWL in MPS rats，which may be related to its effects in inhibiting the activation of spinal dorsal horn microglia and reducing the expression of BDNF.

The effectiveness and safety of Chinese herbal formulas on skin photoaging: A protocol for systematic review and meta-analysis.

BACKGROUND: Skin photoaging (SP) is a complex and complicated process of skin characteristic changes caused by excessive sunlight. Wrinkles, looseness, coarseness, and increase or loss of pigment are the main clinical manifestations of the disease. The pathogenesis of SP mainly involving oxidative stress, inflammatory reaction, immune dysregulation and DNA damage, and so on. In recent years, traditional Chinese medicine, as an significant form of complementary and alternative medicine, has attracted the more and more attention within the field of health care and indicated a desirable effect on SP. Chinese herbal formula (CHF) is an essential part of traditional Chinese medicine interventions, and the number of clinical trails on SP treated by CHFs have shown a growing trend. Therefore, we developed this systematic review and meta-analysis protocol to assess the effectiveness and safety of CHFs in the therapy of SP, so as to provide reliable evidence-based evidence for clinical decision-making. METHODS: A overall literature retrieval will be carried out in 9 electronic journal database. We will include randomized controlled trials (RCTs) on CHFs alone or combined with routine western medicine measures in the treatment of SP. The outcomes we focused on are consists of symptom score (skin relaxation, telangiectasia, pore coarseness, pigmentation, etc), total effective rate, and adverse reactions. Meta-analysis will be performed using Stata 13.0 software. Literature retrieval and screening, data extraction, risk of bias assessment of RCTs, evidence confidence rating by grading of recommendations assessment, development, and evaluation method and methodological quality assessment of systematic review by assessment of multiple systematic reviews-2 will be conducted independently by 2 reviewers, and disagreements will be resolved through discussion or judged by a third senior reviewer. RESULTS: This systematic review and meta-analysis will pool the proof of RCTs on SP treated by CHFs alone or combined with conventional western medicine treatments. The findings of this study will be presented at relevant conferences and submitted to peer-reviewed journals for publication. CONCLUSION: We expect that the results of this systematic review will provide comprehensive and reliable evidence for clinicians and policy makers. REGISTRATION NUMBER: INPLASY 2020120005.

[Effect of electroacupuncture on expressions of Lyn and Syk in mast cells of subcutaneous loose connective tissue in rats with urticarial].

OBJECTIVE: To observe the effect of electroacupuncture (EA) preconditioning on the expressions of tyrosine kinase Lyn and spleen tyrosine kinase (Syk) in mast cells of subcutaneous loose connective tissue in the rats with urticaria and explore the potential biological mechanism of EA in the intervention of urticaria. METHODS: A total of 32 SD rats were randomized into a blank group, a model group, an EA group and a positive medication group, 8 rats in each one. Except of the blank group, the passive cutaneous anaphylaxis (PCA) was adopted to prepare the model of urticaria in the rats of the rest three groups. In the EA group, EA was applied to bilateral "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36), with disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in current intensity, once daily, for 20 min each time, consecutively for 7 days. In the positive medication group, loratadine (1 mg•kg-1•d-1) was for intragastric administration, once daily, consecutively for 7 days. The samples were collected for index detection 30 min after PCA antigen challenge in the rats of each group. Spectrophotometer was adopted to determine the effusion quantity of Evans blue in the allergized site of skin. HE staining was used to observe the morphological changes in the allergized site of skin. Toluidine blue staining was provided to observe mast cell degranulation in subcutaneous loose connective tissue in the allergized site of skin. Immunohistochemistry was applied to determine the protein expressions of Lyn and Syk during degranulation of mast cells. RESULTS: In the rats of the odel group, the eipdermis of allergized site was thickening, cells were disorganized in hierarchy and inflammatory cells were infiltrated largely in the dermis. In the positive medication group and the EA group, the epidermis was getting thin, cell arrangement was clear and the inflammatory cell infiltration was obviously alleviated as compared with the model group. Compared with the blank group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all increased in the model group (P<0.01). Compared with the model group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all reduced in the EA group and the positive medication group (P<0.01). Compared with the positive medication group, the degranulation rate of mast cells was increased significantly in the EA group (P<0.01). CONCLUSION: Electroacupuncture at "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36) reduces vascular permeability and gives play to the role of anti-allergy by the way of regulating and controlling the degranulation of mast cells in the rats with urticaria and the effect mechanism of electroacupuncture may be related to the inhibition of protein expressions of Lyn and Syk in mast cells.

[Electroacupuncture improves cutaneous allergic reaction by inhibiting degranulation of intrape-ritoneal mast cells, MAPK signaling and inflammatory factor levels in urticaria rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on degranulation of intraperitoneal mast cells (MCs) and expression of mitogen-activated protein kinase (MAPK) signaling related proteins, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in urticaria rats, so as to reveal its mechanisms underlying improvement of urticaria. METHODS: Thirty-two SD rats were randomly divided into control，model，EA and medication groups (n＝8 in each group). The urticaria model was established by using passive cutaneous anaphylaxis (PCA) reaction method. EA (2 Hz /15 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Quchi "(LI11) and "Xuehai"(SP10) for 20 min，once daily for 7 consecutive days before antigen attack. Rats of the medication group were treated by gavage of Loratadine(1 mg•kg－1•d－1)for 7 days. The diameter of cutaneous Evan's blue spots was measured to evaluate the severity of PCA. Intraperitoneal fluid smears were prepared to observe the degranulation state of MCs. The contents of TNF-α and IL-6 in the intraperitoneal fluid were detected by ELISA, and the expression of extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, c-Jun N-terminal kinase (JNK), p-JNK, P38MAPK and p-P38MAPK of the acquired intraperitoneal MCs was detected by Western blot. RESULTS: The diameter of cutaneous Evan's blue spot was significantly increased in the model group than that in the control group (P<0.01), and considerably decreased in both EA and medication groups compared with the model group(P<0.01). After modeling，the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of ERK, p-ERK, JNK, p-JNK, P38MAPK and p-P38MAPK were remarkably increased in the mo-del group than those in the control group (P<0.01, P<0.05). After the treatment, the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of p-ERK, JNK, p-JNK and p-P38MAPK were obviously decreased in both EA and medication groups relevant to the model group (P<0.01, P<0.05), while no significant changes were found in the expression of ERK in both EA and medication groups, and P38MAPK in the EA group. Compared with the model and EA groups, expression levels of P38MAPK were down-regulated in the medication group (P<0.05). CONCLUSION: EA can reduce skin allergic reaction in rats with urticaria, which may be related to its effects in inhibiting the degranulation of intraperitoneal MCs, down-regulating the expression of MAPK signaling-related proteins and the level of pro-inflammatory factors TNF-α and IL-6 in intraperitoneal MCs.

[A Meta-analysis of acupuncture therapy for chronic urticaria].

OBJECTIVE: To evaluate the efficacy of acupuncture therapy in the treatment of chronic urticaria. METHODS: Randomized controlled trials of acupuncture treatment of chronic urticaria published from January of 2008 to October of 2018 were searched from databases of CNKI, VIP, Wanfang, CBM, PubMed, Embase and Cochrane Library by using key words "urticaria or urticarias" "acupuncture therapy" "acupuncture treatment" and "randomized controlled trial (RCT)". The retrieved RCT papers were input into EndNote X8 document management software for screening the qualified ones, followed by evaluating the quality of the screened papers. The data extraction table was designed, and the information and data needed for the research were accurately extracted. Finally, Review Manager 5.3 software was used to analyze the included literature. RESULTS: A total of 1 095 articles were retrieved, and 16 RCTs were included finally. The Meta-analysis showed that the total therapeutic efficacy was OR＝3.85, 95% CI [2.61, 5.69], P<0.000 01. The diamonds for comparison between the acupuncture and control (medication) groups in the "Feng Tuan"(urticaria) number, itching, onset duration, swelling degree and scratch test scores were on the right side of the invalid line of forest maps. The recurrence rate (reported in 4 papers) was lower in the acupuncture group than in the control group (OR＝0.28, 95% CI being [0.14, 0.55], P＝0.000 3). CONCLUSION: Acupuncture treatment is significantly better than antihistamine administration alone in the treatment of chronic urticaria, with lower recurrence rate. High-quality clinical studies with rigorous design and large sample sizes are needed for providing more reliable evidence.

[Effect of pre-acupuncture intervention on serum IgE and cutaneous phosphorylated tyrosine-protein kinase expression in rats with urticaria].

OBJECTIVE: To observe the effect of acupuncture on serum IgE level, the degranulation of mast cells, the release of histamine and serotonin, and the expressions of phosphorylated tyrosine-protein kinase Lyn and Syk (p-Lyn, p-Syk) in skin tissue in rats with urticaria, as well as analyze the mechanism of acupuncture in the prevention and the treatment of urticaria. METHODS: SD male rats were randomly divided into normal control, model control, medication and acupuncture groups (n＝10 in each group). The anti-ovalbumin serum was used to establish urticaria model. Rats of the medication group received gastric lavage of Loratadine (0.1 mg/100 g). In the acupuncture group, bilateral "Xuehai" (SP10) and "Quchi" (LI11) were punctured perpendicularly, about 2 to 4 mm in depth, and the needles were retained for 30 min. The treatment was given consecutively for 14 days in the two treatment groups. H．E. staining was adopted to observe the morphological changes of skin tissue, ELISA to determine the total IgE level in serum, the toluidine blue staining to observe the degranulation of mast cells in local skin tissue and the immunohistochemistry to determine the expressions of histamine and serotonin as well as the the expressions of p-Lyn and p-Syk. RESULTS: Compared with the normal control group, the epidermis of the model control group was significantly thickened, the dermis was swollen, the inflammatory infiltration of small vessels was serious and the mast cells were swollen and deformed, with blurred edge and exfoliated granules. Additionally, in the model control group, the serum IgE level was significantly higher (P<0.05), the expressions of histamine and serotonin, as well as p-Lyn and p-Syk proteins in local skin tissue were significantly increased (P<0.05). Compared with the model control group, in the acupuncture group and the medication group, the epidermis was slightly thickened, the dermis got slightly edema, the inflammatory cells were presented occasionally and the degranulation of mast cells were reduced. Besides, the serum IgE level was significantly decreased (P<0.05), the expressions of histamine and serotonin, as well as the p-Lyn and p-Syk were significantly reduced (P<0.05). There was no significant difference in each index between the acupuncture group and the medication group (P>0.05). CONCLUSION: Acupuncture at LI11 and SP10 is applicable in the treatment of urticaria. This therapy inhibits the type Ⅰ hypersensitivity and the mast cell degranulation, which may be related to the regulation of p-Lyn and p-Syk protein expressions in the locus coeruleus skin tissue.

[Effect of deep and shallow electroacupuncture stimulation at "Huantiao"(GB30) on expression of phosphorylated-p38 and phosphorylated-p53 proteins and apoptosis in dorsal root ganglia in sciatic nerve injury rats].

OBJECTIVE: To observe the effect of deep electroacupuncture (EA) stimulation at "Huantiao"(GB30) on hindlimb motor function and expression of p38 mitogen-activated protein kinase (p38 MAPK ) and p53 proteins in dorsal root ganglia (DRG) in rats with chronic constrictive injury (CCI) of sciatic nerve. METHODS: Forty-eight SD rats (half male and half female) were randomly divided into control, model, shallow EA (SEA) stimulation and deep EA (DEA) stimulation groups (n=12 in each group). The CCI model was constructed by implanting a silicone tube close to the sciatic nerve of the left hind limb. For DEA group and SEA group, filiform acupuncture needles were inserted into GB30 about 12-14 mm deep and 5-8 mm deep (monitored by using a high-frequency ultrasound device), respectively, followed by electrical stimulation (2 Hz/100 Hz, 1 mA) using an EA stimulator. The intervention was conducted for 15 min every time, once daily for 14 days. The sciatic nerve function index (SFI) calculated to assess the motor function status. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expression levels of phosphorylated-p38 MAPK (p-p38) and phosphorylated-tumor protein p53 (p-p53) in DRGs of L4-L5 on the affected side were observed by immunohistochemical staining. RESULTS: Following modeling, the SFI were significantly decreased (P<0.01), and the expression levels of p-p38 and p-p53 proteins of L4-L5 DRGs were considerably increased in the model group (P<0.05). After the intervention, the SFI were obviously increased, and the expression levels of p-p38 and p-p53 proteins notably down-regulated in both DEA and SEA groups relevant to the model group (P<0.01, P<0.05). The therapeutic effect of DEA was significantly superior to that of SEA in raising SFI and down-regulating expression le-vels of p-p38 and p-p53 proteins (P<0.01, P<0.05). H.E. staining showed disordered arrangement of the sciatic nerve fibers and myelin, disaggregation of the myelin and axons with deformity and vacuolation in some of them and with an increase of Schwann cells in the model group, which was relatively milder in both DEA and SEA groups. CONCLUSION: Both DEA and SEA at GB30 can obviously improve the motor function in CCI rats, which may be associated with its function in down-regulating the expression of p-p38 and p-p53 proteins in L4-L5 DRGs, restraining p38 MAPK signaling. The therapeutic effect of DEA is evidently better than that of SEA.

[Lower frequency electroacupuncture is better in promoting recovery of limb locomotion in rats with sciatic nerve injury by reducing local inflammatory reaction].

OBJECTIVE: To observe the effect of different frequencies (2 Hz, 100 Hz) of electroacupuncture (EA) on limb locomotion and the expression of inflammatory factors IL-1ß, IL-6, TNF-α in sciatic nerve, and nuclear factor kappa B (NF-κB) in lumber(L)4-L5of spinal cord in rats with sciatic nerve injury (SNI), so as to reveal its mechanisms underlying improvement of SNI. METHODS: A total of 48 SD rats (half male and half female) were equally divided into blank control, model, low frequency (2 Hz) EA and high frequency (100 Hz) EA groups. The SNI model was established by clamping the spinal nerve. EA intervention (2 Hz, 100 Hz, 1 mA), starting on the 8th day after modeling, was applied to "Huantiao" (GB30) on the injured side for 15 min, once daily for 14 consecutive days. The sciatic function index (SFI) was calculated to assess the injured hindlimb recovery with reference to BAIN's and colleagues' methods. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expressions of IL-1ß, IL-6 and TNF-α in the sciatic nerve tissue were detected by immunohistochemistry, and the expression of NF-κB in the spinal cord was detected by using Western blot. RESULTS: After modeling, the SFI level on day 8 was significantly decreased in the model group (P<0.01), and no significant differences were found among the model, low frequency EA and high frequency EA groups before the EA intervention (P>0.05). Following the treatment (at the 22nd day), the SFI values of both low frequency EA and high frequency EA groups were significantly increased (P<0.01), suggesting an improvement of the limb motor function, and the SFI of the low frequency EA group was notably higher than that of the high frequency EA group (P<0.01). In comparison with the blank control group, the expression levels of IL-1ß, IL-6, TNF-α in the sciatic nerve and NF-κB protein in the spinal cord were significantly up-regulated (P<0.05). Following EA intervention, the increased expression levels of IL-1ß, IL-6, TNF-α and NF-κB proteins were significantly down-regulated in both low frequency EA and high frequency EA groups (P<0.05), and the therapeutic effect of low frequency EA was markedly superior to that of high frequency EA in down-regulating the expression levels of IL-1ß, IL-6, TNF-α and NF-κB protein (P<0.05). H.E. staining showed increase of Schwann cells in number, cellular swelling, and disintegration of the axons and myelin sheath, and appearance of vacuolar degeneration in the model group, which was relatively milder in both low frequency EA and high frequency EA groups, particularly in the low frequency EA group. CONCLUSION: EA of GB30 at 2 Hz and 100 Hz can promote the recovery of hindlimb motor function in SNI rats, which is probably related to its function in inhibiting the inflammatory response, and facilitating the repair of the damaged sciatic nerve. 2 Hz EA is better than 100 Hz EA in the therapeutic effect.

[A new attempt of re-mapping acupoint atlas in the rat].

OBJECTIVE: To try to re-map an acupoint atlas in vector mode in the experimental rat by using a new-generation drawing tool. METHODS: A total of 5 SD rats were used in the present study. In reference to the current acupoint locations and anatomical structure of rats as well as those of the human body, an acupoint atlas having a stereoscopic mode was re-mapped by using Adobe photoshop/illustrator CS6 imaging processing and drawing system. RESULTS: According to the skeletal structure and body surface landmarks, an acupoint atlas consisting of Shuigou (GV26), Baihui (GV20), Tianmen (i.e. Cuanzhu, BL2), Erjian (MA-H6), Dazhui (GV14), Feishu (BL13), Xinshu (BL15), Geshu (BL17), Jizhong (GV6), Pishu (BL20), Shenshu (BL23), Houhui (SI3), Huantiao (GB30), Houhai, Yanglingquan (GB34), Housanli (ST36), Zhaohai (KI6), Sanyinjiao (SP6), Genduan point, Shenmai (BL62), Taichong (LR3), Zhijian (foot), Yongquan (KI1), Guanyuan (CV4), Xiqian, Weijian, Shenque (CV8), Zhongwan (CV12), Qiansanli (LI10), Waiguan (SJ5), Neiguan (PC6), Quchi (CV6), Zhoujie (EX-UE1), Hegu (LI4), Zhijian (hand), Houxi (SI3), Shenmen (HT7), Taiyuan (LU9), Shaohai (HT3), Chize (LU5), Danzhong (CV17), and Chengjiang (CV24) which have been positioned in plane graph mode by Professor HUA Xing-bang and colleagues in 1991. The newly re-mapped acupoint atlas displayed more clearly in the anatomic locations relevant to the traditional one. CONCLUSION: A new acupoint atlas with 3 dimension image mode is accomplished in the rat, being more convenient and applicable for researchers to edit the relevant graph materials in paper writing, and playing a possibly useful guidance for the standardization of acupoint selection in experimental acupuncture researches.

[A Meta-analysis on Acupuncture and Moxibustion Treatment of Schizophrenia].

OBJECTIVE: To evaluate the clinical efficacy and security of acupuncture therapy combined with conventional antipsychotics in the treatment of schizophrenia. METHODS: We first comprehensively retrieved papers about acupuncture or moxibustion treatment of schizophrenia published from the time of establishment of the database to April of 2017 in Chinese databases such as Chinese National Knowledge Infrastructure(CNKI), Chinese Science and Technology Periodical Database (VIP), Wanfang, and Chinese BioMedical Literature (CBM) by using keywords "acupuncture""moxibustion""acupuncture ＋moxibustion""acupuncture ＋medication""electroacupuncture""filiform needle""scalp-acupuncture""auricular acupuncture"， etc. Then, according to the principles and methods of evidence-based medicine, papers conformed to the included standards were enrolled to Meta-analysis by using Review Manager 5.3 software. RESULTS: A total of 618 studies were identified, with only 17 articles (containing 1 542 patients) conformed to our included standards and analyzed finally. In regard to the total effective rate of acupuncture or moxibustion for schizophrenia, the combined OR＝3.09, 95% CI (2.31, 4.13), the diamond was on the right side of the vertical line (suggesting being effective), OR checked by Z test (Z＝7.61, P<0.000 01). In regard to the untoward effect, the combined OR＝0.34, 95% CI (0.16, 0.75), the diamond was on the left side of the vertical line (suggesting less adverse effects), OR checked by Z test (Z＝2.67，P＝0.008). Sensitivity analysis: the combined standardized mean difference (SMD)＝－0.61, 95% CI(－0.88, －0.34), the diamond was on the left side of the vertical line (suggesting having more profits than simple medication), SMD checked by Z test (Z＝4.39, P<0.000 1)．. CONCLUSION: Acupuncture and moxibustion combined with conventional antipsychotics is significantly better than that of antipsychotics alone in the treatment of schizophrenia and has less side effects. But, high quality clinical studies with rigorous experimental design and large sample size are needed to provide more reliable evidence.

[Anti-apoptotic Mechanism of Deep Electroacupuncture at "Huantiao" (GB 30) in Repair of the Injured Sciatic Nerve in Rats].

OBJECTIVE: To observe the effect of deep electroacupuncture (EA) at "Huantiao" (GB 30) on functional and pathological changes of the damaged sciatic nerve and apoptosis-related factors in lumbar dorsal root ganglia (DRGs) in rats, so as to study its mechanisms underlying relieving sciatica. METHODS: Forty-eight SD rats were randomly divided into normal, mo-del, deep EA (DEA) and shallow EA (SEA) groups (n＝12 in each group)．The sciatic nerve injury model was established by silicone tube extrusion of the sciatic nerve stem. For DEA group, acupuncture needles were inserted into GB 30 about 8 mm deep to induce nerve impulse under the guidance of high-frequency ultrasound, and the needles were inserted into GB 30 about 3-5 mm in the SEA group. The EA treatment was applied to bilateral GB 30 for 20 min beginning from the 15th day on after modeling, once daily for 14 days. The sciatic nerve function index (SFI) and motor nerve conduction velocity (MNCV) were recorded and calculated. Pathological changes of the sciatic nerve were displayed by H. E. staining. The cell apoptosis and expression of PI 3 K and AKT proteins in lumbar 4-5 DRGs were detected by TUNEL staining, and immunohistochemistry, respectively. RESULTS: In comparison with the normal group, the SFI and MNCV were significantly decreased (P<0.01，P<0.05), the number of neuronal apoptosis and the expression of PI 3 K and AKT in L 4-L 5 DRGs were significantly increased in the model group (P<0.05). Following the treatment, the SFI, MNCV, and the number of neuronal apoptosis were reversed compared with the model group (P<0.01, P<0.05), both PI 3 K and AKT expression levels were significantly up-regulated in both DEA and SEA groups (P<0.05). The therapeutic effects were significantly better in the DEA group than in the SEA group in down-regulating cell apoptosis number and in up-regulating SFI, MNCV, and PI 3 K and AKT protein expression on day 28 of modeling (P<0.01，P<0.05). CONCLUSION: Deep EA at GB 30 can activate PI 3 K-AKT signaling pathway, inhibit the apoptosis of nerve cells in DRGs after sciatic nerve injury in rats, which may contribute to its effect in improving neurological impairment.

Feishu Acupuncture Inhibits Acetylcholine Synthesis and Restores Muscarinic Acetylcholine Receptor M2 Expression in the Lung When Treating Allergic Asthma.

Acupuncture was proven beneficial in treating allergic inflammation. We aimed to explore the regulation underlying the effects of acupuncture on Feishu, an acupoint most commonly used in the acupuncture therapy for respiratory diseases, with respect to the system of sympathetic nerve neurotransmitter acetylcholine (Ach). Male Wistar rats were randomly grouping. No treatment was taken in the normal group. Allergic asthma was induced using ovalbumin on the model, Feishu acupuncture, and sham acupuncture groups; then control or acupuncture treatment lasting for 3 weeks was performed. Bronchoalveolar lavage fluid (BALF) from the four groups was examined. And pulmonary tissues were subjected to histological analysis with H&E staining; besides, immunofluorescent staining, quantitative PCR, and western blot were used to detect synthetase (ChAT) and Ach hydrolase (AchE), and its muscarinic receptors (mAchRs) M1-M3. There was inflammatory infiltration in the lung upon allergic asthma, which was alleviated by the Feishu acupuncture. The eosinophilic granulocytes, neutrophils, and lymphocytes in BALF from the Feishu acupuncture group were all significantly decreased compared with those of the model and sham acupuncture groups. The specific acupuncture on Feishu upon allergic asthma put down the pulmonary expression of ChAT, repaired at the level of gene expression the pulmonary expression of mAchR M1, and restored the pulmonary expression of mAchR M2 (especially in the bronchiolar epithelium) which has a role in inhibiting Ach release; while sham acupuncture had no effect. These results confirmed the therapeutic effects of Feishu acupuncture on allergic asthma, suggesting that the mechanisms may involve suppression of the Ach signal both from its synthesis and during its release.

[Meta-analysis on the Therapeutic Effect of Acupuncture at Meridian Sinew for Spastic Paralysis After Stroke].

OBJECTIVE: To review systematically the clinical effects of spastic paralysis after stroke treated with acupuncture at meridian sinew ("Jingjin", musculotendon). METHODS: "Meridian sinew" "stroke" and "spasm" were taken as the key words to retrieve from the Chinese National Knowledge Infrastracture Database (CNKI), Chongqing VIP Chinese Science and Technology Periodical Database (VIP), Chinese Biomedical Library (CBM), Wanfang Data, PubMed and the Cochrane Library. The Cochrane"risk of bias" tool was used to conduct the methodological quality evaluation to the literature. RevMan 5.3 software was adopted for Meta-analysis. RESULTS: Totally, 13 papers were included, with 820 patients involved. In reference to Cochrane Reviewers' Handbook 5.0.2, the randomized controlled trial (RCT) risk of bias was assessed and it was unclear for all of the 13 papers. The results of Meta-analysis showed that the clinical effect was improved with acupuncture at meridian sinew as compared with normal acupuncture technique[①total effective rate:OR=3.86, 95% CI (2.67,5.57), Z=7.20, P<0.00001; ②modified Ashworth spasm scale:OR=4.54, 95% CI (2.91,7.10), Z=6.64, P<0.00001; ③evaluation of limb motor function with Fugl-Meyer score:MD=4.18, 95% CI (-0.59,8.94), Z=1.72, P=0.09>0.05]. The publication bias of included papers was not obvious and therefore it could be neglected in the impact on the combined effect size. CONCLUSIONS: Acupuncture at meridian sinew is effective in the treatment of spastic paralysis after stroke. The total clinical effect and the improvement in muscular tone with acupuncture at meridian sinew are better than those with normal acupuncture technique. The quality of the included literature is not high generally. Hence, it is necessary to have more clinical studies with high-quality and strict design.

[Development of Researches on Acupuncture Treatment of Peripheral Nerve Injury].

Peripheral nerve injury is a common clinical disease. Acupuncture therapy has been demonstrated to be effective in improving nerve injury in clinical practice, but its underlying mechanisms in prompting tissue repair basically remain unknown. In the present paper, the authors reviewed some descriptions of traditional Chinese medicine on peripheral nerve injury and treatment, and recent development of researches on acupuncture treatment of it in both clinical practice and animal studies. Clinical trials demonstrated that acupuncture treatment can relieve nerve injury induced pain, ameliorate both sensory and motor functions. Experimental studies showed that acupuncture stimulation may promote nerve repair by reducing desquamation of medullary sheath of nerve fibers, inhibiting apoptosis of nerve cells, and up-regulating expression of myelin basic protein, Slit-1 protein and gene, etc. In addition, acupuncture intervention may also improve the microenvironment of neural regeneration including increase of the proliferation and differentiation of Schwann cells and release of various types of neurotrophic factors. However, its mechanisms underlying accelerating rehabilitation of peripheral nerve injury need being researched further.

Moxibustion regulates inflammatory mediators and colonic mucosal barrier in ulcerative colitis rats.

AIM: To observe the efficacy and mechanism of grain-sized moxibustion at different acupoints in a rat model of ulcerative colitis (UC). METHODS: Sprague-Dawley rats were randomly divided into control, UC model, grain-sized moxibustion at a single acupoint (CV 12), grain-sized moxibustion at two acupoints (CV 12 and CV 4), grain-sized moxibustion at three acupoints (CV 12, CV 4, and ST 36), and medication groups (n = 8/group). The UC model was established by enema of trinitrobenzene sulfonic acid. Direct moxibustion was used once a day for 7 d. Disease activity index (DAI) was evaluated before and after the treatment. Morphologic changes of intestinal tissue were observed under an optical microscope. The expression of tumor necrosis factor (TNF)-α and p38 mitogen-activated protein kinase (p38MAPK) in colonic tissue was detected using Western blot, and the levels of occludin and zonula occludens-1 (ZO-1) mRNAs were detected using reverse transcription PCR. RESULTS: Compared with the control group, the intestinal mucosae were incomplete in the model group, glandular structures were irregular, and submucosae were edematous, hyperemic, and infiltrated with inflammatory cells. The DAI scores and expression of TNF-α and p38MAPK were increased significantly in the model group compared to controls (Ps < 0.01), while the mRNA levels of occludin and ZO-1 were reduced significantly (Ps < 0.01). Compared with the model group, colonic mucosa and the arrangement of glands were complete and regular in the treatment groups. DAI scores and the expression of TNF-α and p38MAPK were reduced significantly in moxibustion groups compared to controls (Ps < 0.01), while the mRNA levels of occludin and ZO-1 were increased significantly (Ps < 0.01). The improvements in the above indices in the three acupoints group and the medication group were superior to those in the single and two acupoints groups (all P < 0.05). CONCLUSION: Reduction of TNF-α and p38MAPK and increased expression of occludin and ZO-1 in colonic tissue represent a potential mechanism for improved intestinal mucosal tissue repair with grain-sized moxibustion.