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Objectives: To observe the effect of electroacupuncture (EA) on urodynamics and Raf/MEK/ERK signaling pathway in spine cord tissue of rats after suprasacral spinal cord injury (SSCI), so as to explore its possible mechanism in improving bladder function in rats with detrusor hyperreflexia after SSCI., Methods: Female SD rats were randomly divided into blank, sham operation, model, EA and EA+PD98059 groups, with 12 rats in each group. Thorax (T) 10 spinal cord transection was performed by surgery. Rats in the EA group were given EA (10 Hz/50 Hz, 20 min) at "Ciliao" (BL32), "Zhongji" (CV3), "Sanyinjiao" (SP6) and "Dazhui" (GV14) once daily for 7 d. Rats of the EA+PD98059 group received intraperitoneal injection of PD98059 (5 mg/kg) 2 h before EA intervention. The urodyna-mics was used to measure the base pressure, leak point pressure, maximum pressure, maximum capacity and comp-liance of bladder, and the morphology of bladder detrusor tissue was observed with HE staining. The TUNEL staining was used to detect the cell apoptosis of the spinal cord tissue. The expression levels of exchange protein directly activated by cAMP 2 (Epac2), Rap, phosphorylated rapidly accelerated fibrosarcoma (p-Raf), phosphorylated mitogen-activated extracellular signal-regulated kinase (p-MEK), phosphorylated extracellular signal regulated kinase 1 and 2 (p-ERK1/2), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax) were determined by Western blot., Results: Compared with the sham operation group, the base pressure, leak point pressure and maximum pressure of bladder were significantly increased ( P <0.01), the maximum bladder capacity and bladder compliance were decreased ( P <0.01), the cell apoptosis rate of spinal cord tissue was increased ( P <0.01), and the expression levels of Epac2, Rap, p-Raf, p-MEK, p-ERK1/2, and Bcl-2 protein in spinal cord tissue were decreased ( P <0.01), while the expression level of Bax protein was increased ( P <0.01) in the model group. After the treatment and compared with the model group, the base pressure, leak point pressure and maximum pressure of bladder, the cell apoptosis rate of spinal cord tissue, the expression level of Bax protein were decreased ( P <0.05) in the EA group, while the maximum bladder capacity and bladder compliance, the expression levels of Epac2, Rap, p-Raf, p-MEK, p-ERK1/2, and Bcl-2 protein in spinal cord tissue were all increased ( P <0.05, P <0.01). In comparison with the EA group, the base pressure, leak point pressure and maximum pressure of bladder, the cell apoptosis rate, the expression level of Bax protein were significantly increased ( P <0.05), whereas the maximum bladder capacity, bladder compliance, and the expression levels of p-MEK, p-ERK1/2, and Bcl-2 protein were decreased ( P <0.05) in the EA+PD98059 group. Results of HE staining showed disordered transitional epithelial cells and destroyed lamina propria in bladder detrusor tissue, with the infiltration of monocytes in the model group, which was obviously milder in both EA and EA+PD98059 groups, especially in the EA group., Conclusions: EA can improve the bladder function in detrusor hyperreflexia rats after SSCI, which may be related to its effect in up-regulating Epac2 and Rap, activating the Raf-MEK-ERK pathway, and reducing the cell apoptosis of spinal cord tissue.

Objective: To observe the effect of moxibustion on the indicators of autophagy and apoptosis in the synovium of toes of rats with adjuvant-induced arthritis (AA), so as to explore the underlying mechanism of moxibustion in the treatment of rheumatoid arthritis., Methods: Forty-five SD rats were randomly divided into the blank control group, model group, moxibustion group, methotrexate group and rapamycin group, with 9 rats in each group. The rat model of AA was established by injecting Freund's complete adjuvant. Rats in the moxibustion group received moxibustion treatment at "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, once a day. The methotrexate group was given methotrexate intragastrically (0.35 mg/kg) twice a week. The rapamycin group was given rapamycin by intraperitoneal injection (1 mg/kg), once every other day. The toe volume of the left hind limb was measured by the toe volume measuring instrument after 3-day modeling and 3-week intervention respectively. The contents of interlukin(IL)-1 and tumor necrosis factor(TNF)-α in serum were detected by ELISA. The autophagosomes of synovial cells of the toe joint were observed under transmission electron microscope. The expressions of mammalian target of rapamycin(mTOR)C1, p-mTORC1, Caspase-3, Fas and FasL in synovial tissue were detected by Western blot., Results: Under transmission electron microscope, the model group showed decreased autophagosomes in synovial tissues, but the moxibustion, methotrexate, and rapamycin groups showed increased autophagosomes. Compared with the blank control group, the toe volume, the contents of IL-1 and TNF-α in serum and the expression of p-mTORC1 protein in synovial tissue were significantly increased ( P <0.01, P <0.001), while the expressions of Caspase-3, Fas and FasL proteins in synovial tissue were significantly decreased ( P <0.05, P <0.01) in the model group. Compared with the model group, the toe volume, the contents of IL-1 and TNF-α in the serum, and expression of p-mTORC1 protein were significantly decreased ( P <0.05, P <0.01, P <0.001) in the moxibustion group and the methotrexate group, while the expression of Caspase-3, Fas and FasL proteins in synovial tissue in the moxibustion group and the methotrexate group, the expression of Caspase-3 in the rapamycin group were significantly increased ( P <0.05)., Conclusion: Moxibustion can improve joint swelling in AA rats and decrease the contents of serum IL-1 and TNF-α. The mechanism may be related to regulating the expressions of p-mTORC1, Caspase-3, Fas and FasL proteins, and promoting autophagy and apoptosis of synovial cells.

Objective: To observe the effect of moxibustion on AMP-activated protein kinase (AMPK)/UNC-51-like kinase 1 (ULK1) signaling pathway in the synovial tissue of toes in rheumatoid arthritis rats, so as to explore the mechanism of mo-xibustion in the treatment of rheumatoid arthritis (RA)., Methods: A total of 45 SD rats were randomly divided into blank, model， moxibustion, methotrexate and rapamycin groups, with 9 rats in each group. RA rat model was established by injection of Freund's complete adjuvant. Moxibustion was applied to "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, once a day for 3 weeks. Methotrexate group was given methotrexate (0.35 mg/kg) by gavage, twice a week for 3 weeks. Rapamycin group was intraperitoneally injected with rapamycin (1 mg/kg)，once every other day for 3 weeks. The toe volume of the left hind limb of rats was measured by the toe volume measuring instrument. The content of AMP in toe synovium was detected by ELISA. The expression of AMPK and VPS34 protein in toe synovium was detected by Western blot.The expression of ULK1 and Atg13 mRNA in toe synovium was detected by RT-PCR., Results: Compared with the blank group, the volume of toe in the model group was increased ( P <0.01)，while the content of AMP, the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly decreased( P <0.01).Compared with the model group, the volume of toe in the moxibustion，methotre-xate and rapamycin groups was decreased ( P <0.05); the content of AMP, the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly increased( P <0.05) in the moxibustion group; the content of AMP, the expression of VPS34 protein, the expression of Atg13 mRNA were significantly increased( P <0.05) in the methotrexate group; the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly increased ( P <0.05, P <0.01) in the rapamycin group. Compared with the moxibustion group, the expression of AMPK protein in the methotrexate group and the content of AMP in the rapamycin group were significantly decreased ( P <0.05)., Conclusion: Moxibustion can improve joint swelling in RA rats, and the mechanism may be related to promoting the activity of AMPK/ULK1 signaling pathway.

Objective: To observe the effect of electroacupuncture (EA) of "Zusanli"(ST36) and "Guanyuan"(CV4) on the apoptosis rate of synoviocytes and protein expression of Fas, FasL and Caspase-3 in synovial tissue of adjuvant-induced arthritis (AIA) rats, so as to explore its mechanisms underlying improvement of rheumatoid arthritis （RA）., Methods: A total of 24 rats were randomly divided into normal, model, medication and EA groups, with 6 rats in each group. The AIA model was established by injection of complete Freund's adjuvant (CFA, 0.1 mL) into the left hindlimb paw. The rats in the medication group received intraperitoneal injection of 0.35 mg/kg of methotrexate, twice a week for 4 weeks. The rats in the EA group received EA stimulation of ST36 and CV4 (20 Hz/50 Hz, 1 mA) for 20 min, 6 times a week for 4 weeks. The left hind paw volume was measured using a paw volume meter, and histopathological changes of synovial tissue were observed by light microscope after H.E. staining. The serum contents of tumor necrosis factor-α（TNF-α） and interleukin-1 （IL-1） were measured by ELISA. The apoptosis of synoviocytes was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL), and the expression of apoptosis-associated proteins Fas, FasL and Caspase-3 in synovium were detected by Western blot., Results: Compared with the normal group, the left hind paw volume from day 3 to 24 after administration of CFA, serum IL-1 and TNF-α contents were significantly increased ( P <0.01), while the expressions of Fas, FasL and Caspase-3 proteins and apoptotic rate of synoviocytes were significantly decreased in the model group ( P <0.01). In comparison with the model group, the paw volume from day 17 to 24 after modeling, the serum IL-1 and TNF-α contents were significantly reduced ( P <0.01), while the apoptotic rate of synoviocytes, expressions of Fas protein in both medication and EA groups, Caspase-3 protein in the acupuncture group and FasL protein in the medication group were increased ( P <0.01, P <0.05). Compared with the medication group, the expression of FasL protein was decreased in EA group ( P <0.05). H.E. stain showed obvious hyperplasia of the synovial lining layer, and disordered arrangement of synovial cells, with edema and enlargement in some cells in the model group, which was relatively milder in both medication and EA groups., Conclusion: EA of ST36 and CV4 can promote the apoptosis of synoviocytes and the expressions of Fas and FasL proteins in AIA rats, which may contribute to its role in relieving synovitis through activating Fas/FasL signaling.

Objective: To observe the effects of electroacupuncture (EA) on autophagosomes, unc-51-like autophage activating kinase 1 (ULK1), Beclin1, and microtubule-associated protein light chain 3 (LC3) expression, and synoviocyte ultrastructure in the synovial tissues of rheumatoid arthritis (RA) rats, and to explore the mechanism of EA in regulating the proliferation of synoviocytes via the autophagy pathway., Methods: The SD rats were randomly divided into a normal group, a model group, a methotrexate (MTX) group, and an EA group, with 6 rats in each group. Following RA modeling with Freund's complete adjuvant, rats in the MTX group were treated with intragastric administration of 0.35 mg/kg MTX, twice a week, for 4 weeks, while those in the EA group received 20-min EA stimulation at "Zusanli" (ST36) and "Guanyuan" (CV4), once per day, for 4 weeks, with an interval of one day between weeks. The rat left hind toe volume was measured using the toe volume measuring instrument. HE staining was conducted for detecting the morphology of rat synovial tissues, followed by the observation of autophagosomes under the transmission electron microscope. The levels of serum interleukin (IL)-1 and tumor necrosis factor (TNF)-α were assayed by ELISA, and the protein expression levels of ULK1, Beclin1, and LC3 were detected by Western blot., Results: Compared with the normal group, the left hind toe volume of the model group increased significantly ( P <0.01), while serum IL-1 and TNF-α ( P <0.01), synovial ULK1, Beclin1, and LC3 protein expression ( P <0.01, P <0.05) up-regulated. HE stain and electron microscope showed obvious synovial hyperplasia, and doublemembrane autophagosomes scattering in the synoviocytes. The comparison with the model group showed that MTX and EA remarkably decreased the left hind toe volume ( P <0.01), serum IL-1 and TNF-α ( P <0.01), down-regulated the protein expression levels of ULK1 and LC3 in synovial tissues ( P <0.05, P <0.01), and inhibited the synovial hyperplasia, with no obvious autophagosomes observed in the synoviocytes. The protein expression of ULK1 in the EA group was significantly lower than that in the MTX group ( P <0.01)., Conclusion: EA alleviates the joint swelling and synoviocyte injury of RA rats possibly by regulating the expression of ULK1, LC3, and Beclin1 and inhibiting the synoviocyte autophagy and proliferation.

Objective: To observe the effect of electroacupuncture (EA) on urodynamics of neurogenic bladder and pituitary adenylate cyclase activating peptide(PACAP)/cyclic adenosine monophosphate (cAMP)/protein kinase A(PKA) signaling pathway in detrusor tissue of rats after suprasacral spinal cord injury (SCI), so as to explore its possible mechanism in improving detrusor hyperreflexia bladder function after shock stage of suprasacral SCI., Methods: Female SD rats were randomly divided into control, sham operation, model and EA groups, with 12 rats in each group. T10 spinal cord transection (SCT) was performed by surgery. Rats in the EA group were given EA (10 Hz/50 Hz, 20 min) at "Ciliao" (BL32), "Zhongji" (CV3) and "Sanyinjiao" (SP6) once daily for 7 days. After the intervention, urodynamics testing was detected in each group. HE staining was used to observe the morphological changes of bladder detrusor. The protein and mRNA expression of PACAP38 in detrusor was detected by immunohistochemistry, Western blot and real-time quantitive PCR, respectively. The contents of cAMP and PKA were determined by ELISA., Results: Compared with the control and sham operation groups, the maximum bladder capacity and bladder com-pliance, and the protein and mRNA expression of PACAP38, and the contents of cAMP and PKA of the model group were significantly decreased ( P <0.01, P <0.05), while the base pressure and leakage point pressure of bladder were significantly increased ( P <0.01). After EA intervention, the above indexes were all reversed in the EA group relevant to those of the model group ( P <0.01, P <0.05)., Conclusion: EA at BL32, CV3 and SP6 can improve the bladder function in rats with bladder detrusor hyperreflexia after SCI, which may be related to its effect in activating the PACAP/cAMP/PKA signaling pathway in detrusor tissue.

Objective: To observe the effect of electroacupuncture (EA) stimulation of "Xiaohai" (SI 8) and "Xiajuxu" (ST 39) on serum TNF-α and duodenal high mobility group protein B 1 (HMGB 1) levels in rats with duodenal ulcer (DU), so as to analyze its underlying mechanism in improving DU., Methods: Forty SD rats were randomly divided into normal control, model, Xiaohai (SI 8) and Xiajuxu (ST 39) groups (n=10 in each group). The DU model was established by intramuscular injection of 10% Cysteamine hydrochloride (300 mg/kg), followed by taking water containing 1% Cysteamine hydrochloride for 48 h. EA (10 Hz/50 Hz, 1-3 mA) was applied to bilateral SI 8 and ST 39 for 30 min, once a day for 10 days. The DU score (0-5 score) was evaluated according to Moraes' method. Serum TNF-α content was determined by ELISA and duodenal HMGB 1 expression was measured by immunohistochemistry., Results: Compared to the normal control group, the DU score, serum TNF-α content, and duodenal HMGB 1 expression level of the model group were significantly higher (P<0.01). Following EA intervention, the increased DU score, serum TNF-α content, and duodenal HMGB 1 expression level were considerably down-regulated in the two EA groups in comparison with the model group (P<0.01, P<0.05) except HMGB 1 in the SI 8 group (P>0.05), and the effect of EA of "Xiajuxu" (ST 39), the lower-He point, was remarkably superior to that of "Xiaohai" (SI 8) in reducing DU score and TNF-α content (P<0.01)., Conclusion: EA stimulation of SI 8 and ST 39 can improve duodenal ulceration in DU rats, which may be related to its effect in down-regulating serum TNF-α and duodenal HMGB 1 expression. The therapeutic effect of ST 39 is markedly better than SI 8 in relieving duodenal ulceration.

Objective: To observe the effect of electroacupuncture (EA) stimulation on the expression of metalloprotei- nase-9 (MMP-9) and tissue inhibitor-1 (TIMP-1) in the lung tissue in rats with chronic obstructive pulmonary disease (COPD), so as to reveal its mechanisms in protecting the lung tissue., Methods: Forty male SD rats were equally randomized into control, model, medication and EA groups. The COPD model was established by smoke-fumigation method (passive smoking in a closed box) and endotracheal injection of lipopolysaccharide (200 microg/100 microL) for 30 days. Rats of the medication group were treated by i.p. of dexamethasone acetate injection (2.0 mg/kg), once daily for 22 days. EA was applied to LU 9, ST 36, ST 40 and KI 3 for 20 min, once daily for 22 days. Pathological changes of the pulmonary tissue were observed under optical microscope after hematoxylin and eosin (H.E.) staining. The expression of pulmonary MMP-9 and TIMP-1 was assayed by immunohistochemistry., Results: H.E. staining showed pulmonary diffuse edema, severe inflammatory cell infiltration, and increase of the numbers of goblet cells, proliferation of fibroblasts, etc. in the model group which were relatively milder in the medication and EA groups. The expression levels of pulmonary MMP-9 and TIMP-1 proteins were significantly higher in the model group than in the control group (P<0.05), and were considerably down-regulated in both EA and medication groups in comparison with the model group (P<0.05). There were no significant differences between the EA and medication groups in MMP-9 and TIMP-1 expression levels (P>0.05)., Conclusion: EA can effectively suppress the increased expression of pulmonary MMP-9 and TIMP-1 in COPD rats, which may contribute to its effect in improving COPD-induced pathological changes.

Objective: To observe the effect of acupoint-catgut-embedding therapy on blood lipid and insulin levels in type I diabetes mellitus rats;s as to reveal its mechanism underlying improvement of pancreatic functions., Methods: Thirty SD rats were divided into normal, model and treatment group. The type I diabetes model was established by feeding the rats with high-fat diet and intraperitodneal injection of streptozotocin (STZ). Catgut-embedment was performed in acupoint "Zhongwan" (CV 12)-"Xiawan" (CV 10), "Yishu"-"Ganshu" (BL 18), etc., once every 20 days, twice altogether. Fasting blood glucose (FBG) content was assayed by glucose oxidase method; glycosylated hemoglobin A 1 c (HbA 1 c) assayed by ELISA, and serum fasting C-peptide(FC-p) deterriined by chemiluminescence method; and the triglyceride (TG), total cholesterol (TC), and serum insulin (INS) levels were detected by enzymatic method, separately. The rat's pancreas tissues were fixed with 4% paraformaldehyde and cut into sections (4 Im in thickness) and stained with H. E. method., Results: After modeling, the levels of FBG, HbA 1 c, INS and FC-p, TG, TC were significantly increased in the model group in comparison with those of the normal group (P<0.05), while INS sensitive. index was apparently decreased in the model group (P<0.05). Following catgut-embedding treatment, changes of the above-mentioned indexes were all remarkably reversed (P<0.05), and H.E. staining showed an improvement of the injured pancreatic tissue and islet cells., Conclusion: Catgut-embedding therapy can reduce fasting blood glucose and insulin resistance, and improve lipid metabolism in type I diabetic rats, which may contribute to its effect in protecting pancreatic islet cells.

Objective: To observe the effect of moxibustion on epidermal growth factor receptor (EGFR) and phosphorylated extracellular signal regulated kinase 1/2(p-ERK1/2) protein expression in gastric ulcer (GU) rats so as to reveal its mechanisms underlying improving GU., Methods: Fifty SD rats were randomly allocated to control,model, medication ("Sijunzi" Decoction), non-acupoint moxibustion (non-acup-moxi), and acup-moxi groups (n=10 in each group). The GU model was established by intragastric perfusion of absolute alcohol. Rats of the control group were treated by gavage of distilled water. Rats of the medication group were treated by administration of "Sijunzi" Decoction (8 mL x kg(-1) d(-1)),twice a day for 8 days. Moxibustion intervention was applied to bilateral "Zusanli" (ST 36),"Zhongwan" (CV 12),and "Pishu" (BL 20), "Weishu" (BL 21) alternatively for 30 min, once daily for 8 days. The animals' ulcer index (UI) was assessed by Guth's method, and gastric mucosal pathological changes were observed under light microscope following H. E. staining. The expression of gastric EGFR was detected by immunohistochemistry and that of phosphorylated ERK1/2 (p-ERK1/2) protein determined by Western blot., Results: Compared with the control group, the UI, gastric EGFR and p-ERK1/2 protein expression levels were significantly increased in the model group(P<0.01, P<0.05); whereas in comparison with the model group, the UI was notably decreased in the medication, non-acup-moxi and acup-moxi groups (P<0.05, P<0.01), and EGFR and p-ERK1/2 protein expression levels were further up-regulated in the three treatment groups (P<0.01). The effects of both medication and acup-moxi groups were obviously superior to those of the non-acup-moxi group (P<0.01, P<0.05). No significant differences were found between the medication and acup-moxi groups in the expression levels of EGFR and p-ERK1/2 proteins (P>0.05). Results of H.E. staining showed that alcohol-induced gastric mucosal injury as breakage, exfoliation, inflammatory cell infiltration, etc. was milder in both medication and acup-moxi groups following the treatment., Conclusion: Acupoint-moxibustion has a role in relieving alcohol induced gastric mucosal injury in the rat, which may be closely associated with its effects in up-regulating activities of the EGFR/ERK signal transduction pathway.