OBJECTIVE: To observe the effects of electroacupuncture (EA) at changbing fang (prescription for intestinal disease) on autophagy of colonic cells and gut microbiota in rats with ulcerative colitis (UC), and to explore the mechanism of EA in the treatment of UC. METHODS: Thirty-two SD male rats were randomly divided into a control group, a model group, an EA group and a sham-EA group, with 8 rats in each group. Except the control group, the UC rat model was established by free drinking of 5% dextran sulfate sodium solution for 7 days in the other groups. In the EA group, changbing fang was adopted, in which, EA was applied at "Tianshu" (ST25) and "Shangjuxu" (ST37), at disperse-dense wave and frequency of 10 Hz/50 Hz, for 20 min in each intervention. In the sham-EA group, shallow transcutaneous puncture was performed at the sites, 5 mm away from the points as the EA group, with the same parameters as the EA group. The intervention was delivered once daily for 3 consecutive days. The body weight was measured daily and the disease activity index (DAI) score was calculated before and after intervention. After intervention completion, the colon length was measured. Using HE staining, the colon morphology was observed and the score of colonic pathology was assessed. With ELISA adopted, the contents of tumor necrosis factor (TNF-α), interleukin (IL)-1β, IL-2 and IL-10 in the serum of the rats were detected. The ultrastructure of the colon tissue was observed under electron microscopy. Using Western blotting, the protein expression was detected for microtubule-associated protein 1 light chain 3 (LC3)Ⅱ, LC3Ⅰ, autophagy-related genes (ATG) 5, ATG12, sequestosome 1 (p62), phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-AKT), protein kinase B (AKT), and phosphorylated mammalian target of rapamycin (p-mTOR), mammalian target of rapamycin (mTOR) in the colon tissue. The mRNA expression of PI3K, AKT and m-TOR in the colon tissue was detected by real-time fluorescence quantitative PCR. The 16S rRNA gene sequencing was used to analyze the structure of gut flora in the feces of rats. RESULTS: From day 1 to day 7, compared with the control group, the body weight decreased in the model group, EA group, and SEA group (P<0.05, P<0.01). From day 9 to day 10, the EA group showed an increase in body weight compared with the model group and SEA group (P<0.05, P<0.01). Before intervention, the DAI score in the model group, EA group, and SEA group was higher than the score of the control group, respectively (P<0.01). After intervention, the DAI score in the EA group was reduced compared with the model group and SEA group (P<0.01). Compared with the control group, in the model group, the colon length of rats was shorter (P<0.01); it showed the distorted crypts, thinner mucosal layer, reduced goblet cells, inflammatory cell infiltration and the disarranged histological structure; and the pathological score of the colon tissue increased (P<0.01); the serum contents of TNF-α and IL-1β increased (P<0.01), and those of IL-2 and IL-10 decreased (P<0.01). The structure of colon epithelial cells was disarranged, with cilia pelt off, and a large number of vacuoles in the cytoplasm; the mitochondria were swollen, with unclear structure and cristae partially disappeared; and few autophagosomes were observed. The value of LC3Ⅱ/LC3Ⅰand the protein expression of ATG5 and ATG12 in the colon tissues were reduced (P<0.01), the protein expression of p62 and PI3K, and the values of p-AKT/AKT, and p-mTOR/mTOR increased (P<0.01), and mRNA expression of PI3K, AKT and mTOR was elevated (P<0.01). The indexes of Chao1, Ace and Shannon decreased (P<0.01). At the phylum level, the relative abundance of Firmicutes decreased (P<0.05), that of Bacteroidetes and Proteobacteria increased (P<0.05, P<0.01). At the genus level, the relevant abundance of Lactobacillus decreased (P<0.05), while that of Lachnospiraceae_NK4A136_group and Phascolarctobacterium increased (P<0.01, P<0.05 ). Compared with the model group and SEA group, in the EA group, the colon length increased (P<0.01), the infiltration of inflammatory cells was reduced, the arrangement of intestinal epithelial cells was arranged regularly, with a small amount of shedding, and the pathological score of the colon tissue decreased (P<0.01). The serum contents of TNF-α and IL-1β decreased (P<0.01), and those of IL-2 and IL-10 increased (P<0.01). The colonic epithelial cells were arranged relatively, the morphology of organelles was basically normal, and autophagosomes were visible. The value of LC3Ⅱ/LC3Ⅰand the protein expression of ATG5 and ATG12 in colon tissue increased (P<0.01, P<0.05), the protein expression of p62 and PI3K, and the values of p-AKT/AKT, and p-mTOR/mTOR decreased (P<0.01); and mRNA expression of PI3K, AKT, m-TOR was reduced (P<0.01). The indexes of Chao1, Ace and Shannon increased (P<0.01). At the phylum level, the relative abundance of Firmicutes increased (P<0.01), while that of Bacteroidetes decreased (P<0.01). At the genus level, the relative abundance of Lactobacillus increased (P<0.05), whereas that of Lachnospiraceae_NK4A136_group decreased (P<0.01). When compared with the model group, the relative abundance of Proteobacteria decreased (P<0.05), and that of Phascolarctobacterium was reduced (P<0.05) in the EA group. CONCLUSION EA at changbingfang alleviates UC symptoms probably through inhibiting the PI3K/AKT/mTOR signaling pathway to regulate colonic autophagy and improve the intestinal flora.
Animals ; Electroacupuncture ; Colitis, Ulcerative/metabolism* ; Male ; Rats ; Gastrointestinal Microbiome ; Rats, Sprague-Dawley ; Colon/metabolism* ; Humans ; Autophagy ; Acupuncture Points ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-10/genetics*

Objective: To investigate the neural connections between Shenmen (HT7)-heart and the brain by observing the tracing viruses co-labeled brain nuclear groups after injection of the pseudorabies viruses (PRV), the reverse transsynaptic virus tracer carrying different fluorescent protein genes, into the myocardium and Shenmen (HT7) point, respectively.Methods: Pseudorabies virus 531 (PRV531) carrying the green fluorescent protein gene and pseudorabies virus 724 (PRV724) carrying the red fluorescent protein gene were injected into the left ventricular wall and Shenmen (HT7) point area of the left forelimb of six C57BL/6 mice, respectively. After 120 h, whole brain tissue was extracted under 4％ paraformaldehyde perfusion to prepare brain sections. Neuronal co-labeling with the tracing viruses was observed under fluorescence microscopy. Results: Co-labeled signals from the mouse ventricular wall and Shenmen (HT7) point region were found at all levels of the mouse central nervous areas, such as the cerebral cortex, hypothalamus, midbrain, pons, and medulla oblongata. The number of co-labeled neurons was higher in the primary motor area, the hypothalamic paraventricular nucleus, the subceruleus nucleus, and the paramedian reticular nucleus. Conclusion: There is a neural connection between Shenmen (HT7), the heart, and the brain, which may be most closely related to the autonomic nervous system.

Objective: To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid (NMDA) receptor subtype 2B (NR2B) in the hippocampus of rheumatoid arthritis (RA) rats, and to explore the analgesic mechanisms of moxibustion in RA treatment. Methods: Sixty male Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, a moxibustion + NMDA receptor antagonist (AP-5) group, and a moxibustion + NMDA receptor agonist (NMDA) group, with 12 rats in each group. Except for the normal group, rats in the other four groups were treated with complete Freund's adjuvant in a windy, cold, and damp environment to replicate RA models. Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu (BL23) and Zusanli (ST36), and the two points were used alternately. After intraperitoneal injection of AP-5 or NMDA, rats in the moxibustion + AP-5 group and the moxibustion + NMDA group received the same moxibustion intervention as in the moxibustion group, once a day for 15 d. The thermal withdrawal latency (TWL) of rats in each group was detected before and after modeling and after the 15-day intervention. After the 15-day intervention, hematoxylin-eosin staining was performed to observe the pathological changes in knee joints. The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus; Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B. Results: The synovial tissue was proliferated, the synovial lining was significantly thickened, the pannus was formed, and the cartilage and bone tissues were significantly damaged in the model group. After intervention, the pathological morphology of the knee joints in the moxibustion group, the moxibustion + AP-5 group, and the moxibustion + NMDA group was significantly improved, and the improvement in the moxibustion + AP-5 group was more notable than that in the moxibustion + NMDA group. Compared with the normal group, the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group (P<0.01). Compared with the model group, the TWL of each intervention group was significantly increased (P<0.01 or P<0.05), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased (P<0.01). Compared with the moxibustion group, the TWL was significantly increased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion + AP-5 group (P<0.01); the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion + NMDA group (P<0.01). Conclusion: Moxibustion reduces hyperalgesia in RA inflammatory rats. The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.

OBJECTIVETo observe the regional homogeneity (ReHo) of resting-state brain function in the healthy subjects of gastric distention treated with acupuncture at the back- and front- points of the stomach, Weishu (BL 21) and Zhongwan (CV 12) and the correlation with gastric motility so as to explore the mechanism on the central integration of the front- and back- points of the stomach.

METHODSThe crossover test design was adopted. Twenty-four healthy subjects were assigned to a Weishu group, a Zhongwan group and a combined-point group separately, 8 cases in each one in each of the three times. Totally, 24 subjects were included in each group. Under the water load condition, the subjects received acupuncture at Weishu (BL 21), Zhongwan (CV 12) and the combined Weishu (BL 21) and Zhongwan (CV 12). Before and after each acupuncture, the resting-state brain functional magnetic resonance imaging (fMRI) scan and electrogastrogram (EGG) test were applied. The ReHo value was calculated in the collected fMRI imaging data. The changes in ReHo values were analyzed and compared before and after acupuncture in each group, as well as among the groups. The gastric motility was analyzed before and after acupuncture. Additionally, the correlative analysis was conducted between the gastric motility and ReHo changes before and after acupuncture.

RESULTS(1) After acupuncture, EGG amplitudes in the subjects of each group were lower remarkably as compared with those before acupuncture (all <0.01). The EGG frequencies were not different significantly as compared with those before acupuncture (all >0.05). The EGG amplitudes in the Weishu group and the Zhongwan group were higher than those in the combined-point group (both <0.05). (2) As compared with the conditions before acupuncture, acupuncture at the combined front- and the back- points as well as Weishu (BL 21) and Zhongwan (CV 12) separately all induced the changes in the brain ReHo. Acupuncture at the combined front- and the back- points significantly increased Reho values in the right inferior temporal gyrus, the left thalamus, the precuneus and the posterior cingulate gyrus (all <0.05) and remarkably reduced the ReHo values in the the middle temporal gyrus of the right temporal pole, sulcus calcarinus and precuneus (all <0.05). Compared with the single point groups, acupuncture at the combined front- and the back- points induced the increase of ReHo value in the posterior cingulate gyrus and the decrease of ReHo in the temporal pole (all <0.05). (3) The correlative analysis showed that the changes in the ReHo values in the posterior cingulate gyrus, the thalamus and the precuneus were positively correlated to the changes of the gastric motility amplitudes. The changes in the ReHo values in the temporal pole was negatively correlated to the changes of the gastric motility amplitudes.

CONCLUSIONAcupuncture at the combined back- and front- points of the stomach, as well as acupuncture at single Weishu (BL 21) and Zhongwan (CV 12) induce the ReHo changes in the different brain regions. Acupuncture at the combined back- and front- points of the stomach may induce the ReHo changes in some new brain regions as compared with the acupuncture at the single point. The thalamus, the posterior cingulate gyrus and the precuneus may be the the important integrated brain regions for acupuncture at the back- and the front- points in regulating the gastric motility. The effects of acupuncture at the back- and the front- points for the regulation of the gastric motility are closely related to the thalamus, the limbic system and the default network of the brain regions.

Acupuncture Points ; Acupuncture Therapy ; Brain ; diagnostic imaging ; physiology ; Cross-Over Studies ; Electroencephalography ; Gastrointestinal Motility ; Healthy Volunteers ; Humans ; Magnetic Resonance Imaging ; Stomach ; physiopathology

Objective To discuss the influence of Tanshinone ⅡA on the tight junction protein of intestinal mucosal epithelial cells in rat severe septic models. Methods Seventy-five Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group and Tanshinone ⅡA injection high (20 mg/kg), medium (10 mg/kg) and low (5 mg/kg) dose groups, each group 15 rats. Sepsis rat models were established by cecal ligation and puncture (CLP) method, in sham operation group, only switched abdominal surgery was performed without CLP. In Tanshinone ⅡA injection groups, different doses of Tanshinone ⅡA were injected intraperitoneally after modeling for 10 minutes and 6 hours; in sham operation and model groups, equal volume of normal saline was injected intraperitoneally at the same times as above. After operation, 3 L/kg of normal saline was injected into the caudal vein in all rats for fluid resuscitation.Twelve hours after operation, the rats were killed, the abdominal lymph nodes, liver, spleen and kidney tissues were taken for bacterial culture and calculating the rate of bacterial translocation; under microscope, the histopathological changes of ileum mucosal tissues were examined and Chiu scoring was carried out; TdT-mediated dUTP nick end labeling (TUNEL) was applied to detect the ileum mucosal epithelial cell apoptosis and calculating the index (AI);fluorescence immunoassay and Western Blot methods were used to measure the contents and protein expression levels of tight junction protein, junctional adhesion molecule-1 (JAM), Claudin-1, Zonula occludens-1 (ZO-1), Occludin, c-Fos and Tryptase. Results ① In bacterial cultures of abdominal lymph node, liver, spleen and kidney, the positive rate of mesenteric lymph node was the highest, followed by liver and spleen, mainly Escherichia coli, Proteus mirabilis, etc. The highest positive rate of bacterial culture was in model group (38.8%), followed by low dose of Tanshinone ⅡA injection group (35.0%), and the lowest was 16.6% in high dose Tanshinone ⅡA injection group, the differences being statistically significant in comparisons between any pair of groups (all P < 0.05). ② Pathological examination showed that the pathological changes of ileum mucosa were obvious and the Chiu score (4.17±0.98 vs. 0) and AI (11.70±2.87 vs. 2.17±0.80) in model group were significantly higher than those in sham group (all P < 0.05); with the increase of dosage of Tanshinone ⅡA injection, the pathological changes of rat ileum mucosa were improved gradually, the Chiu score and AI were decreased gradually, and the degrees of decrease in high dose Tanshinone ⅡA group were more significant than those in model group (Chiu score: 1.12±0.79 vs. 4.17±0.98, AI: 3.65±1.98 vs. 11.70±2.87, both P < 0.05).③ Immunofluorescence staining showed that the positive staining of protein JAM, ZO-1 and c-Fos were all green in color, Claudin-1, Occludin and Tryptase were all red in color, the localizations of all of them were in the cytoplasm, the protein expression of JAM, Claudin-1, ZO-1, Occludin from strong to weak in turn were Sham group, high, medium, low dose Tanshinone ⅡA group and model group, the expression of c-Fos, Tryptase from strong to weak in turn were model group, low, medium, high dose Tanshinone ⅡA group and Sham group. ④ Western Blot showed that the expressions of ileum tissue JAM, Claudin-1, ZO-1 and Occludin in model group were all significantly lower than those of the sham group, while the expressions of c-Fos, Tryptase were obviously higher than those of the sham group, with the increase of dosage of Tanshinone ⅡA, the expressions of JAM, Claudin-1, ZO-1 and Occludin were increased gradually and the protein expressions of c-Fos and Tryptase were gradually decreased, and the changes in high dosage group of Tanshinone ⅡA were more significant than those in low and moderate groups [JAM (gray value): 25.39±1.82 vs. 12.41±1.34, 19.45±1.66, Claudin-1 (gray value): 28.44±1.56 vs.17.26±1.46, 21.23±1.34, ZO-1 (gray value): 28.84±1.59 vs. 16.45±1.21, 24.22±1.46, Occludin (gray value): 25.49±1.63 vs. 13.34±1.45, 19.45±1.37, c-Fos (gray value):15.76±1.36 vs. 27.84±1.36, 21.22±1.73, Tryptase (gray value): 14.44±1.41 vs. 28.14±1.38, 22.32±1.57], all the above comparisons of different dosage groups were statistically significant (all P < 0.05). Conclusion Tanshinone ⅡA injection may improve intestinal wall structure and reduce bacterial translocation by improving the intestinal mucosal tight junction protein in sepsis model rats, and this effect is positively correlated to Tanshinone ⅡA dosage.

Objective To compare the characteristics,complications and outcomes of the modified lapa-roscopic Swenson(MLSw)and laparoscopic Soave(LS)procedures for children with short-segment Hirschsprung disease(HD). Methods Seventy-seven pediatric patients with HD who underwent surgery from March 2007 to December 2016 were enrolled in this retrospective study. Twenty-six patients were treated with LS and 51 cases un-derwent MLSw. The preoperative,operative and postoperative data was collected,with follow-up periods ranging from 12 to 48 months. The perioperative/operative characteristics,postoperative complications,and outcomes were compared between the two groups. Results On average,the patients in the LS group had a longer operating time than that in the MLSw group(P < 0.05). Blood loss was significantly less in the MLSw group than that in the LS group(P < 0.05). There was no significant difference in feeding time between the two groups(P > 0.05). The MLSw group was discharged after a shorter hospitalization time than that in the LS group(P < 0.05). The MLSw group had lower incidences of postoperative complications than those in the LS group in the early postoperative period,with no significant difference in the rate of complications during the late postoperative period was found between the two groups. Conclusions Both LS and MLSw are suitable for treatment of children with short-segment HD. However,the MLSw operation is much simpler,with less operating time,less intraoperative blood loss,shorter hospitalization time and better bowel control in the early postoperative period. We favor this approach because it allows complete removal of the entire original aganglionic bowel,without leaving behind a cuff.

OBJECTIVETo explore the partial action mechanism and the myocardial protective effect differences between electroacupuncture (EA) preconditioning at "Neiguan"(PC 6) and "Taiyuan"(LU 9) in rats with acute myocardial ischemia-reperfusion injury.

METHODSNinety-six Wistar rats were randomly assigned into a sham-operation group, a model group, a Neiguan group and a Taiyuan group, 24 rats in each one. The rats in the Neiguan group and Taiyuan group were treated with EA (2 Hz in frequency, 1 mA in intensity) at "Neiguan" (PC 6) and "Taiyuan" (LU 9) respectively, 20 min per treatment, once a day for consecutive 7 days. The rats in the sham-operation group and model group were treated with immobilization for the same time, and no EA was given. The model of myocardial ischemia-reperfusion injury was established in the model group, Neiguan group and Taiyuan group 24 h after the end of EA, while the rats in the sham-operation group were treated with sham operation (no ligation was made during surgery). The myocardial ischemic size, infarction size, activity of protein kinase C (PKC) and expression of aquaporin1 (AQP1) in each group were detected.

RESULTSCompared with sham-operation group, the myocardial ischemic size, infarction size, AQP1 expression and PKC activity in the model group were significantly increased (all<0.01); compared with the model group and Taiyuan group, the myocardial ischemic size, infarction size, PKC activity and AQP1 expression were significantly decreased in the Neiguan group (<0.01,<0.05). By Pearson correlation analysis, the changes of AQP1 expression were positively correlated with those of PKC activity after EA preconditioning.

CONCLUSIONSEA preconditioning at "Neiguan" (PC 6) could significantly decrease myocardial AQP1 expression and PKC activity in rats with acute myocardial ischemia-reperfusion injuing, but the effect of EA preconditioning at "Taiyuan"(LU 9) is not obvious; its protective effect is likely to be achieved by inhibiting PKC activity and AQP1 expression.

OBJECTIVETo compare the difference in the clinical therapeutic effects of the regulation of subhealthy condition between the mild moxibustion on alternate days and Chinese herbal medicine.

METHODSSeventy-two participants of subhealthy condition were assigned into a mild moxibustion group (36 cases) and a medication group (36 cases) according to random number table. In the mild moxibustion group, the intervention of mild moxibustion with moxa stick was applied to Guanyuan (CV 4), Zusanli (ST 36) and Sanyinjiao (SP 6), once every other day. In the medication group,granules were prescribed for oral administration, three times a day. The treatment was continued for 2 months. Before and after treatment, the levels of IgA, IgM and IgG, SOD (superoxide dismutase), the total score of fatigue scale 14 (FS-14) and the score of nailfold microcirculation were observed separately.

RESULTSCompared with the results before treatment, the immune indices, the total score of FS-14 and the scores of nailfold microcirculation were all improved after treatment in the two groups, indicating the significant differences (<0.05,<0.01). In comparison between the two groups after treatment, the levels of immune globulin, e.g. IgM, IgG and SOD in the mild moxibustion group were higher in tendency than those in the medication group, but without significant differences (all>0.05). The total score of FS-14 after treatment in the mild moxibustion group was lower than that in the medication group (<0.05). After treatment, in the mild moxibustion group, for the nailfold microcirculation, the flow state score, the periloop score and the total score were all reduced more apparently as compared with those in the medication group (<0.01,<0.05).

CONCLUSIONSMild moxibustion with moxa stick on alternate days apparently relieves fatigue state, immune dysfunction and microcirculatory impairment in the subhealthy group. The results are apparently superior to the treatment withgranules.

Objective To investigate the prevalence of feeding intolerance (FI),and to explore the FI within 7 days of ICU admission in association with clinical outcome in critically ill patients.Methods The adult patients from 14 general ICUs in Zhejiang Province with an expected admission to ICU for at least 24h were recruited from March 2014 to August 2014,and all clinical,laboratory,and survival data were prospectively collected.The AGI (acute gastrointestinal injury) grade was daily assessed based on gastrointestinal (GI) symptoms,feeding details and organ dysfunction within the first week of ICU stay.The intra-abdominal pressures (IAP) was measured using AbViser device.Results Of 550 patients enrolled,418 were assessed in GI symptoms and feeding details within 7 days of ICU stay.The mean age and SOFA score were (65.1 ± 18.3) years and (8.96 ±4.10),respectively.Of them,355 patients (84.9％) were under mechanical ventilation support,and 37 (8.85％) received renal replacement therapy.The mean length of time for enteral feeding was (30.8 ±26.2) h,and the prevalence of FI on the 3rd and 7th day of ICU stay accounted for 39.2％ and 25.4％,respectively.Compared to those with FI within 7 days of ICU stay,the patients without FI had higher rate of successively weaning from mechanical ventilation (21.3％ vs.5.7％,P =0.003) and higher rate of withdrawal of vasoactive medication (45.5％ vs.20.0％,P =0.037),as well as lower mortality rate of 28-day (24.4％ vs.38.7％,P =0.004) and 60-day (29.6％ vs.44.3％,P =0.005).In multivariate Cox regression model with adjustment for age,sex,participant center,serum creatinine and lactate,AGI grade on the first day of ICU stay,and comorbidities,the FI within 7 days of ICU stay (x2 ≥ 7.24,P ＜ 0.01) remained to be independent predictors for 60-day mortality.After further adjusted for SOFA score,the FI within 7 days of ICU stay (HR =1.71,95％ CI:1.18-2.49;P =0.006) and AGI grade on the first day of ICU stay (HR =1.33,95 ％ CI:1.07-1.65;P =0.009) could provide independent prognostic values of 60-day mortality.Conclusions There is high rate of FI occurred within 7 days of ICU stay,and is significantly associated with worse outcome.In addition,this study also provides evidence to further support that measurement of gastrointestinal dysfunction could increase value of SOFA score in outcome prediction for the risk of 60-day mortality.

Objective To compare the effects of mild moxibustion at different frequencies on sub-health state, and to discuss the effectiveness and dose-response relationship of moxibustion in treating sub-health state. Method Totally 108 subjects were divided by using random number table into group Ⅰ (36 cases), group Ⅱ (36 cases) and group Ⅲ (36 cases). Same acupoints were selected for the three groups, including Guanyuan (CV3), bilateral Zusanli (ST36), and bilateral Sanyinjiao (SP6). A moxa stick was ignited at one end to treat the corresponding points, 3-4 cm away from the skin, making the topical area hot but without burning pain. Each point was treated for 10 min, till the skin flushed. The 3 groups were respectively treated once a day, once every other day, and once a week, 1 month as a treatment course, totally for 2 courses. The Sub-health Symptoms Scale, immunoglobulins IgA, IgM and IgG, and superoxide dismutase (SOD) were evaluated and statistically analyzed before and after intervention. Result The total effective rate was 87.9% in group Ⅰ, 84.8% in group Ⅱ, and 34.3% in group Ⅲ. The paired comparisons showed that there was no significant difference between group Ⅰ and Ⅱ (P>0.05), while group Ⅰ and group Ⅱ were both significantly different from group Ⅲ (P<0.05). After intervention, the levels of IgA, IgM, IgG, and SOD were significantly increased in group Ⅰ and Ⅱ (P<0.05), but the increases in group Ⅲ were statistically insignificant (P>0.05). Conclusion The therapeutic efficacy of mild moxibustion for sub-health is related to the treatment frequency, and frequencies at once a day and once every other day both can produce significant efficacies, which are superior to that of moxibustion at once a week.