Objective:To investigate the regulatory effects of acupuncture and moxibustion on chromatin remodeling complex core catalytic subunit of Brahma-related gene 1(Brg1),histone deacetylase(HDAC)3,HDAC9,and males absent on the first(MOF)in the colon tissue of rats with Crohn disease(CD). Methods:Using the random number table method,60 male Sprague-Dawley rats were divided into 5 groups,including a normal group,a model group,an acupuncture group,a medicinal cake-insulated moxibustion group,and an acupuncture-moxibustion group,with 12 rats in each group.CD rat models were prepared using 2,4,6-trinitrobenzene sulfonic acid(TNBS)in all groups except the normal group.The normal and model groups received no interventions.In the acupuncture group,rats were intervened with acupuncture at bilateral Zusanli(ST36)and Shangjuxu(ST37),20 min/session,once a day.The medicinal cake-insulated moxibustion group received medicinal cake-insulated moxibustion at Qihai(CV6)and bilateral Tianshu(ST25)with 2 cones per point per session,once a day.The acupuncture-moxibustion group received both acupuncture and moxibustion interventions simultaneously.Each intervention was performed for 10 consecutive days.Observations included general condition,disease activity,macroscopic damage,and pathological changes in the rat's colon tissue.Real-time fluorescence quantitative polymerase chain reaction was used to measure the mRNA expression of chromatin remodeling-related enzymes Brg1,HDAC3,and HDAC9,while Western blotting detected the protein expression of Brg1,HDAC3,HDAC9,and MOF in rat's colon tissue. Results:The model group showed significantly increased diarrhea score,occult blood score,macroscopic damage score of colon tissue,and colon macroscopic damage index(CMDI)score,as well as elevated mRNA expression levels of HDAC3 and HDAC9,protein expression levels of HDAC3,HDAC9,and MOF,and decreased mRNA and protein expression levels of Brg1 compared to the normal group(P<0.01).In contrast,compared to the model group,the diarrhea score,occult blood score,macroscopic damage score,CMDI score,mRNA expression levels of HDAC3 and HDAC9,and protein expression levels of HDAC3,HDAC9,and MOF were significantly reduced(P<0.05 or P<0.01),while the mRNA and protein expression levels of Brg1 were significantly increased(P<0.01)in the acupuncture group,the medicinal cake-insulated moxibustion group,and the acupuncture-moxibustion group. Conclusion:Both medicinal cake-insulated moxibustion and acupuncture,either used alone or in combination,can regulate the abnormal expression of chromatin remodeling-related enzymes Brg1,HDAC3,HDAC9,and MOF in the colon tissue,thus reducing colon inflammation in CD rats.

Objective:To observe the effects of acupuncture and moxibustion on interleukin(IL)-9/IL-9 receptor(IL-9R)in the colon tissue of rats with ulcerative colitis(UC)and investigate the protective mechanism of acupuncture and moxibustion on the intestinal mucosal barrier in UC rats. Methods:Male Sprague-Dawley rats were randomly divided into a normal control(NC)group and a modeling group.UC models were prepared by giving 4%dextran sulfate sodium(DSS)water for 7 d.After the successful construction of the UC rat model,the modeling group was randomly divided into a UC group,a herb-insulated moxibustion(HM)group,and an electroacupuncture(EA)group.HM and EA interventions at bilateral Tianshu(ST25)were performed once a day for 7 d.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in the colon.The serum concentrations of IL-9,IL-6,IL-1β,and hemoglobin-H(HbH)were determined by enzyme-linked immunosorbent assay.The protein expression levels of IL-9,IL-9R,claudin-2,zonula occludens-1(ZO-1),and occludin in the colon tissue were measured by Western blotting or immuno-histochemistry.Immunofluorescence was used to detect the co-expression of PU.1 and CD4 with the IL-9 protein. Results:Compared with the NC group,the colon tissue of UC rats was severely damaged and ulcerated with congestion and edema,and the colonic histopathological score increased significantly(P<0.01).The serum HbH concentration decreased significantly(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β increased(P<0.01).The protein expression of colonic ZO-1 and occludin decreased significantly(P<0.01),while the protein expression of colonic IL-9 and IL-9R increased(P<0.05).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 increased in the colon tissue(P<0.05).Compared with the UC group,the colonic mucosal structures were gradually repaired in both HM group and EA group,and healed ulcers could be observed,the colonic histopathological score decreased significantly(P<0.05).The serum concentration of HbH increased(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β decreased(P<0.05).The protein expression levels of ZO-1 and occludin increased(P<0.05),while the protein expression levels of IL-9 and IL-9R decreased(P<0.01).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 decreased in the colon tissue(P<0.05). Conclusion:Both HM and EA can inhibit the protein expression levels of IL-9 and IL-9R in the UC colon by regulating the transcription factor PU.1,promote the repair of intestinal mucosal barrier,and down-regulate protein contents of proinflammatory factors IL-9,IL-6,and IL-1β in the serum,which may be one of the key mechanisms of acupuncture and moxibustion in reducing the inflammation of UC colonic mucosa and protecting the intestinal mucosal barrier.

Objective: To observe the anti-inflammatory effect, as well as the effect on the expression of microtubule-associated protein light chain 3B (LC3B) and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis (EAT). Methods: Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group. The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction. After the model was prepared, rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group. In the herbal cake-partitioned moxibustion group, moxibustion was alternately applied to two groups of points [Dazhui (GV14)-Mingmen (GV4) and Tiantu (CV22)-Guanyuan (CV4)], and the treatment continued for 30 d. Rats in the normal and model groups were only fixed identically without intervention. Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining; the concentrations of thyroid peroxidase antibodies (TPOAb), thyroglobulin antibodies (TGAb), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay; real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue. Results: There were massive follicular destruction, lymphocytic infiltration, and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group. Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group. Compared with the normal group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were increased in the model rats (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group (P<0.05). Compared with the model group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were reduced in the herbal cake-partitioned moxibustion group (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group (P<0.05). The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion: Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb. This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.

Objective: To observe the effect of herb-partition moxibustion on colonic inflammation in UC rats, and participate in the regulation of immune regulation via NF-κB pathway and STAT3 phosphorylation. Methods: Rats were randomly divided into normal group, the ulcerative colitis group (UC) group and herb-partition moxibustion group, with 8 rats in each group. The rat model of UC was induced by 4％ DSS. After successful modeling, the rats were treated with moxibustion on bilateral Tianshu acupoints (ST25) . Each acupoint was used with 2 Zhuang moxa, 1 time a day, for 7 times. The effect of the herb-partition moxibustion on UC rats was observed by comparing the histopathological and protein concentrations of serum proinflammatory cytokine. Western Blot was used to detect NF-κB pathway and STAT3 activity in colon tissue. Results: Colonic histopathology in the UC group showed that the mucosal epithelium with ulcer formation and obvious inflammatory response. The herb-partition moxibustion could repair colonic epithelial damage and reduce the inflammatory response of colon tissue in UC rats. Compared with the normal group, the serum IL-1β and IL-6 protein concentrations were significantly increased in the UC group (P ＜ 0.05, P ＜ 0.01), and the STAT3 phosphorylation level and protein expression levels of pIκB-α and NF-κB p65 were significantly increased. The protein expression level of IκB-α was significantly decreased. Compared with the UC group, the serum protein concentrations of IL-1β and IL-6 were significantly lower in the herb-partition moxibustion group (P ＜ 0.05), and protein expression level of NF-κB p65 was decreased in the colon tissue. The phosphorylation level of pIκB-α was decreased, while the protein expression level of IκB-α was increased. Conclusion: Herb-partition moxibustion reduced colonic inflammatory response in UC rats by DSS-induced, the underlying mechanism may related to decrease release of proinflammatory cytokines IL-1β and IL-6 via dual inhibition of NF-κB and STAT3 activation.

Objective: To categorize and summarize the clinical and mechanism studies of the past 30 years on the treatment of Hashimoto’s thyroiditis (HT) with moxibustion, moxibustion plus medication, and acupuncture plus medication, etc., and to analyze the current problems. Methods: The clinical and laboratory studies related to the treatment of HT with acupuncture-moxibustion therapies published before June 2015 were retrieved from MEDLINE, Excerpta Medica Database (EMBASE), China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang) and Chongqing VIP Database (CQVIP). Results:Moxibustion, moxibustion plus medication, and acupuncture plus medication can produce certain therapeutic effects in treating HT. Conclusion:The research on the treatment of HT with acupuncture-moxibustion therapies is rather limited in the amount and content. In the future, standardization should be fortified, specific moxibustion research needs deepening, and the action mechanism of moxibustion should be emphasized.

Objective:To investigate the influence of moxibustion products on mitochondrial transmembrane potential (MTP) and mRNA expression of Bax/Bcl-2 in alveolar typeⅡ epithelial A549 cells, and to further explore influence of moxibustion products on the oxidative damage of A549 cells. Methods:Smoke and particles generated by moxibustion were collected using the filter box for gas sampling. The moxa smoke extract (MSE) was diluted sequentially to the final concentrations of 0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL using the cell culture medium, and A549 cells were then intervened by the above MSE solution. Cell MTP was detected by JC-1 staining. Fluorescence quantitative polymerase chain reaction (PCR) was used to detect Bax/Bcl-2 mRNA expression of A549 cells. Results: Compared with cells in the normal control group, MTP was significantly decreased in cells of 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P<0.01); while MTP showed no significant changes in cells of 0.05 mg/mL, 0.1 mg/mL and 0.2 mg/mL MSE intervention groups (P>0.05); compared with cells in 0.05 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P<0.05 ); compared with cells in 0.1 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.4 mg/mL MSE intervention group (P<0.01). Bax mRNA expression of cells in each concentration of MSE intervention group all showed no significant difference compared to that in the normal control group; Bcl-2 mRNA expression of cells was reduced with the increase of MSE intervention concentration. Wherein, Bcl-2 mRNA expressions of cells in 0.4 mg/mL and 0.3 mg/mL MSE intervention groups were significantly reduced compared with that of cells in the normal control group (P<0.05); Bcl-2 mRNA expression of cells in 0.4 mg/mL MSE intervention group was significantly reduced compared to that in 0.05 mg/mL MSE intervention group (P<0.05). Conclusion:Certain higher concentration of moxa smoke could reduce MTP and mRNA expression of the anti-apoptosis gene Bcl-2 in alveolar typeⅡ epithelial A549 cells. Oxidative damage may be the important mechanism of apoptosis caused by the high concentration of moxa smoke solution, and further studies are necessary on the specific mechanisms.

Objective:To observe the effect of moxibustion on learning and memory abilities, corticosterone and glucocorticoid receptor (GR) in subacute aging rats. Methods:Twenty four Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group and a moxibustion group, 8 rats in each group. Rats in the model group and the moxibustion group were subcutaneously injected with 25% D-galactose [125 mg/(kg·bw)] for 40 d continuous; rats in the normal group were injected with saline at the same position for 40 d continuous. Rats in the moxibustion group were given mild moxibustion at bilateral Shenshu (BL 23) at the same time of modeling; rats in the normal group and the model group were only identically grabbed without moxibustion for 40 d. The learning and memory abilities of rats were observed using the Morris water maze at the end of the experiment. Abdominal aorta blood and thymus were collected after water maze experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone level, and immunohistochemical method was used to detect the expression of thymus GR. Results:Compared with the normal group, rats in the model group showed that a significantly longer escape latency time (P<0.01) on the third and the fourth days; number of times crossing the platform in 70 s significantly reduced (P<0.01); activity times in the fourth quadrant significantly decreased (P<0.05); serum corticosterone levels increased (P<0.01); thymus GR expression decreased (P<0.05). Compared with the model group, rats in the moxibustion group showed that the escape latency times were significantly shorter on the third, the fourth and the fifth days (P<0.01,P<0.05); number of times crossing the platform in 70 s significantly increased (P<0.05); activity times in the fourth quadrant significantly increased (P<0.05); serum corticosterone levels decreased (P<0.05); thymus GR expression increased (P<0.05). Conclusion:Moxibustion could improve the learning and memory abilities of subacute aging rats, down-regulate serum corticosterone levels, and increase thymus GR content.

Objective:To evaluate and compare electroacupunctures (EA) with different parameters and moxibustion at different temperatures influencing the activation of mast cells (MC) in Tianshu (ST 25) regions of visceral hyperalgesia model rats. Methods:Rats (except for model group) respectively accepted 1 mA or 3 mA EA or moxibustion at 43 or 4 to℃℃stimulate Tianshu (ST 25) points after randomization of the fifty visceral hyperalgesia model rats, and then were compared with that in model and normal groups. Number, degranulation numbers, degranulation rates in Tianshu (ST 25) regions MC of rats in each group were observed using toluidine blue staining. Abdominal withdrawl reflex (AWR) score was used to evaluate the rat visceral hyperalgesia reactions. Results:Compared with the normal group and the model group, MC numbers (P<0.05,P<0.01,P<0.01,P<0.01), degranulation numbers and degranulation rates (P<0.01,P<0.01, P<0.05,P<0.01) of Tianshu (ST 25) MC in regions tissues in 43℃ and 4℃ moxibustion groups, and 1 mA and 3 mAEA groups all increased significantly. Compared with the model group, AWR scores were significantly lower in 43℃ and 4℃ moxibustion groups, and 1 mA and 3 mAEA groups under the stimulation of 20 mmHg, 40 mmHg, 0 mmHg or 80 mmHg colorectal distension (CRD) (P<0.05 in 1 mA and 3 mA EA groups under the stimulation of 20 mmHg,P<0.01 in the other groups). AWR scores in 43℃ and 4℃moxibustion groups under the stimulation of 20 mmHg, 40 mmHg, 0 mmHg or 80 mmHg CRD were not significantly different from those in the normal group (allP>0.05); AWR scores in 1 mA EA group under the stimulation of 0 mmHg or 80 mmHg were significantly higher than that in the normal group (P<0.01); AWR score in 3 mA EA group under the stimulation of 0 mmHg was significantly higher than that in the normal group (P<0.01), and AWR scores in 3 mA EA group under the stimulation of 20 mmHg or 80 mmHg were also higher than that in the normal group (P<0.05). AWR scores were higher in 1 mA EA group under the stimulation of 40 mmHg or 80 mmHg than that in 4℃ moxibustion group (P<0.05); AWR score was higher in 3 mA EA group under the stimulation of 40 mmHg than that in 4℃ moxibustion group (P<0.05). Conclusion:There are differences among EA of different parameters and moxibustion of different temperatures in activating on Tianshu (ST 25) regions MC of visceral hyperalgesia model rats, as well as in improving the visceral hyperalgesia reaction. The effect of 4℃ moxibustion is the most significant.

Objective: To observe the clinical effect and syndrome scores improvements of herbal cake-partitioned moxibustion (HPM) and ginger-partitioned moxibustion (GPM) in treating ulcerative colitis (UC). Methods:A total of 65 eligible cases were randomly divided into a HPM group (n=32) and a GPM group (n=33) according to their visiting order. Bilateral Tianshu (ST 25) and Dachangshu (BL 25) were selected for the HPM or the GPM treatment once daily, 12 d as a treatment course with a 3-day interval, 6 courses in all. The clinical effect, syndrome scale and Mayo scale were evaluated and compared between the two groups. Results:Of the 65 cases enrolled, 2 cases dropped out in the HPM group and 3 cases dropped out in the GPM group, 30 cases of each group finished the treatment courses. The total effective rate is 93.3% in HPM group and 86.7% in the GPM group, there was no statistically significant difference in the total effective rate between the two groups (P>0.05); there were statistically significant differences between the two groups in score evaluation of lasting time of abdominal pain and frequency of diarrhea, HPM is prior to GPM (P=0.032,P=0.044). There are no statistical significant differences between the two groups in scores evaluation of general symptom, three main symptoms, quality of life (QOL), frequency and severity of abdominal pain, times, and pattern of diarrhea (allP>0.05). There was a statistical significant difference in the improvement of Mayo score between the two groups, and HPM was superior to GPM (P=0.048). Conclusion:HPM and GPM are both promising ways to treat UC, and the total effect is quite similar. HPM is superior to GPM in the improvement of lasting time of abdominal pain and frequency of diarrhea, and also the Mayo score.

OBJECTIVETo observe the effects and safety of clinical common symptoms of active Crohn's disease at the mild and moderate stages treated with acupuncture and moxibustion.

METHODSOne hundred and two patients were randomly divided into an observation group and a control group, 51 cases in each one. Herb-partition moxibustion and acupuncture were used in the observation group. Herbal cakes with(),(),(),(),(),() and() as the main ingredients were used on Tianshu (ST 25), Qihai (CV 6) and Zhongwan (CV 12); acupuncture was used at Zusanli (ST 36), Shangjuxu (ST 37), Sanyinjiao (SP 6), Taixi (KI 3), Gongsun (SP 4) and Taichong (LR 3). Bran-partition moxibustion and shallow acupuncture were applied in the control group. Wheat bran-partition moxibustion in the shape of cake was implemented on the same acupoints as the observation group; shallow acupuncture was used 1~2 cm next to the acupoints. Treatment was given three times a week for 12 weeks, 36 times totally. Clinical effects of eight symptoms were evaluated by TCM symptom score, and the symptoms included abdominal pain (degree, frequency, time), diarrhoea (number, shape and frequency), fatigue, anorexia, borborygmus andflatus, fear of cold, soreness and weakness of waist and knees, tenesmus.

RESULTSAfter 12-week treatment, the eight symptoms were improved apparently (all<0.05), with the scores of abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia in the observation group decreasing more obviously (all<0.01). There was no statistical significance about the other symptom scores between the two groups (all>0.05).

CONCLUSIONSHerb-partition moxibustion combined with acupuncture achieve safety and efficacy for mild and moderate Crohn's disease, and can improve the clinical common symptoms. Furthermore, their effects on abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia are better than those of bran-partition moxibustion and shallow acupuncture.