Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

Objective:To observe the effect of electroacupuncture (EA) on the expression of erythropoie-tin-producing hepatocyte receptor B2 (EphB2) in the cortex around the infracted area of middle cerebral artery occlusion (MCAO) rats at different timing, and to reveal the possible mechanism of acupuncture in the treatment of cerebral ischemia.
Methods:A total of 180 male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, with 45 rats in each group. Rats in each group were further divided into three subgroups: postoperative 3 d, postoperative 14 d and postoperative 21 d groups, with 15 rats in each subgroup. The MCAO model was made by the modified occlusion method. The neurological function score, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, immunohistochemistry assay, immunofluorescence double labeling method and Western blot were used to detect the corresponding indicators.
Results:The neurological impairment of rats was most obvious at postoperative 3 d, and then gradually improved with time, which was more significant in the acupoint group (P<0.05). The change of infarcted volume was consistent with the neurological function impairment. The number of EphB2 positive cells (EphB2+) around the infarcted area was decreased significantly at postoperative 3 d, and then gradually improved with time, which returned to the same level as that in the sham operation group at postoperative 21 d. The increase was most significant in the acupoint group (P<0.05), and the positive cell number was higher than that in the sham operation group (P<0.01). Western blot and immunohistochemistry results were basically consistent. Immunofluorescence displayed that EphB2+ and postsynaptic density-95 positive (PSD-95+) were co-expressed, after the MCAO operation, in the cortical neuron around the infracted area, and the number of co-expressing cells was increased gradually with time, which was most significant in the acupoint group (P<0.05).
Conclusion:Electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function and cerebral infarcted volume ratio of MCAO rats, which may be related to the activation of EphB2 expression in cortex around the infracted area and the promotion of synaptic remodeling.

Objective To assess nutritional risks in children with allergic proctocolitis induced by cow milk protein,and to observe the effects of the feeding strategies for them with different interventions in feeding,and then to identify the best time and the best way to modify feeding strategies according to different individual.Methods The effectiveness of different feeding interventions was retrospectively analyzed after 2 weeks in 171 cases with allergic proctocolitis induced by cow milk protein in Hunan Children's Hospital during the period of Jan.2012 to Jul.2013.Thirty-two cases received breast feeding (18.7％) and 139 cases received non-breast feeding (81.3％).Intervention methods:32cases with breast feeding received the same feeding method;among 139 babies with non-breast feeding group,36 were plus free amino acid (AAF) (21.1％),and 103 were plus the extensively hydrolyzed formula (eHF).The intervention formula changes were observed in 8,12 and 24 weeks,respectively,and nutritional risks in 0,3,6 months were assessed,respectively.Results There were 171 patients totally,male to female ratio was 1.5 to 1.0,and the average age was (4.1 ± 1.3) months.After 2 weeks,147 cases were notably effective,accounting for 86％,and in the breast feeding group 79.2％ of the patients were notably effective,and there was no significance compared with the AAF group and eHF group (P ＞ 0.05).The intervention formulation changes during the following 6 months were as follows:in 8 weeks,30 cases in the breast feeding group breastfeeding continued,2 cases turned to eHF feeding for the lack of breast milk,21 cases were fed with AAF (12.3％),85 cases with eHF (49.7％),and 35 cases with moderately hydrolyzed formula (20.5％) ;in 12 weeks:23 cases had breast feeding (13.5％),16 cases were fed with free amino acid formula powder (9.4％),56 cases with extensively hydrolyzed formula (32.7％),and 76 cases with moderately hydrolyzed formula (44.4％) ;in 24 weeks:21 cases (12.3％) had breast feeding,7 cases were fed with AAF(4.1％),13 cases with moderately hydrolyzed formula(7.6％),and 130 cases with normal formula(76.0％).The mean levels of the nutritional risks were assessed according to STAMP in 0,3,6 months respectively:breast feeding group [(1.69 ± 0.78) scores,(1.50 ± 0.88) scores,(1.53 ± 0.67) scores,P ＞ 0.05] ; AAF group [(1.72 ± 0.78) scores,(1.53 ± 0.88) scores,(1.53 ± 0.65) scores,P ＞ 0.05] ; eHF group [(1.80 ± 0.69) scores,(1.68 ± 0.68) scores,(1.66 ± 0.65) scores,P ＞ 0.05].Conclusions The nutrition risk levels in children with milk protein allergic proctocolitis are low,and different feeding interventions according to tolerance had no impact on the nutritional status ; Breast feeding could effectively avoid cow milk allergy,and symptoms of the children with allergic proctocolitis would be relieved after 2-week intervention,and more than half of children could gain part of immune tolerance after intervention for 3 months or more,so for the children with allergic proctocolitis induced by cow milk protein,routine formula feeding should be stopped after 3 months,and individualized feeding intervention can be carried out according to immune tolerance to relieve the financial burden.

Objectives To explore ventilation induced cytokine production and the role of oxidant stress in lung stretch. Methods Both in vitro and in vivo models of ventilator-induced lung injury (VILI) were used. Alveolar epithelial cells were stretched in vitro to mimic the lung injury in VILI. Rats were ventilated at large tidal volume to produce ventilator-induced lung injury in vivo. A total of 23 inflammatory cytokines were screened with micro gene array in stretched alveolar epithelial cells. Cytokines found to have up-regulated in cells were measured in serum and lung tissue of rats exposed to large tidal volume ventilation. For investigating the intracellular pathway of cytokine up-regulation in VILI, exogenous TNF-α or H2O2 was added to culture media of alveolar epithelial cells. Cytokines were then measured. To explore the role of oxidant stress in VILI, N-acetylcysteine (NAC), as an anti-oxidant, was used in vitro and in vivo. Results We found that transforming growth factor-β1 (TGF-β1 and transforming growth factor-β2 (TGF β2) were up-regulated in stretched alveolar epithelial cells and also in serum of rats with large tidal volume ventilation. Tumor necrosis factor-α (TNF-α) had no effects on TGF-β production in alveolar epithelial cells. Exogenous H2O2, as an oxidant, increased TGF-β production in alveolar epithelial cells. NAC, an anti-oxidant, decreased stretch induced TGF-β production, along with a down-regulation of oxidant injury. NAC also blocked the up-regulation of TGF-β in in vivo model of VILI. Conclusion TGF-β1 and TGF-β2 were up-regulated in VILI. Oxidant injury mediated up-regulation of TGF-β in VILI. NAC, which attenuated oxidant injury and blocked TGF-β up-regulation in VILI, could be a future therapeutic strategy in VILI.

Objective To evaluate the significance of human interleukin-31(IL-31)in the pathogenesis of atopic dermatitis and its correlation with pruritus in patients with atopic dermatitis(AD).Methods Twenty-two children with mild to severe atopic dermatitis and 22 age-matched healthy controls were included in this study.Patients and controls were randomly and equally assigned into stimulation and non-stimulation groups.Venous blood samples were obtained from all participants,peripheral blood mononuclear cells were isolated from these samples and cultured with(stimulation groups)or without(non-stimulation groups)staphylococcal enterotoxin B(SEB)for 24 hours.Then,the mRNA expression of IL-31 on PBMCs was assessed via real-time reverse transcription-PCR.ELISA was used to detect the total serum IgE level in these objects.The severity of AD in patients was rated according to scoring atopic dermatitis(SCORAD).The relationship between the mRNA expression of IL-31 and the level of serum total IgE.severity of atopic dermatitis,and degree of pruritus.was evaluated.Results The expression of IL-31 mRNA on non-stimulated PBMCs from patients was 23.2 folds as high as that from the healthy controls(P＜0.01).The stimulation with SEB upregulated the mRNA expression of IL-31 on PBMCs.and the increase on PBMCs from patients was 20.44 times of that from the controls.The total serum IgE level was 260.05 IU/mL(5.9-1131.01 IU/mL)and 17.7 IU/mL(5-140.7 IU/mL)in the Patients and controls respectively(P＜0.01).There was no significant correlation between the mRNA expression of IL-31 and disease severity or total serum IgE level(r=0.07.0.22respectively.both P＞0.05)in patients witll AD.Condusions IL.3 1 is involved in t11e pathogenesis of AD,which is unlikely to be IgE-dependent.SEB can induce the rapid expression of IL-31 on PBMCs of healthy human,and is an important modulator for the production of IL-31.

Objective To evaluate the capacity of atopy patch test in diagnosis of food allergy in children with atopic dermatitis (AD).Methods Egg and milk,as the most common food allergens among Chinese children,were employed in this study.Skin prick test (SPT) and atopy patch test (APT) with fresh egg and milk were carried out in 68 children with AD.Oral food challenges in an open style were performed to confirm the diagnosis of food allergy.Resuits Of these patients,58(85.3%)were sensitive to egg,40(58.8%)to milk and 34(50.0%) to both.Of 98 patients with positive challenge,47 showed late response,10 immediate reactions.and 41 mixed reactions.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and the agreement with food challenges in diagnosis of egg/milk allergy were 96.6%/67.5%.90.0%/82.1%,98.2%/84.4%,81.8%/63.9% and 95.6%/73.5%,respectively for APT alone,37.9%/30.0%,100%/89.3%,100%/80.0%,21.7%/47.2% and 47.1%/54.4%,respectively for SPT alone.APT was found to be more sensitive in diagnosis of late-phase reactions than SPT (P<0.01).No significant difference was found in the sensitivity between APT alone and the combination of APT and SPT in parallel algorithm for diagnosis of egg or milk allergy (x2=0.509,0.549,both P>0.05) or in the specificity between APT alone and that in serial algorithm( P=1.000;x2=3.514,P>0.05).Conclusions APT is superior to SPT in diagnosis of late responses to food,and the combination of SPT and APT does not facilitate the diagnosis of food or milk allergy compared with APT alone.

Objectives To explore ventilation induced cytokine production and the role of oxidant stress in lung stretch. Methods Both in vitro and in vivo models of ventilator-induced lung injury (VILI) were used. Alveolar epithelial cells were stretched in vitro to mimic the lung injury in VILI. Rats were ventilated at large tidal volume to produce ventilator-induced lung injury in vivo. A total of 23 inflammatory cytokines were screened with micro gene array in stretched alveolar epithelial cells. Cytokines found to have up-regulated in cells were measured in serum and lung tissue of rats exposed to large tidal volume ventilation. For investigating the intracellular pathway of cytokine up-regulation in VILI, exogenous TNF-? or H_ 2O_ 2 was added to culture media of alveolar epithelial cells. Cytokines were then measured. To explore the role of oxidant stress in VILI, N-acetylcysteine (NAC), as an anti-oxidant, was used in vitro and in vivo. Results We found that transforming growth factor-?_1 (TGF-?_1 and transforming growth factor-?_2 (TGF-?_2) were up-regulated in stretched alveolar epithelial cells and also in serum of rats with large tidal volume ventilation. Tumor necrosis factor-? (TNF-?) had no effects on TGF-? production in alveolar epithelial cells. Exogenous H_ 2O_ 2, as an oxidant, increased TGF-? production in alveolar epithelial cells. NAC, an anti-oxidant, decreased stretch induced TGF-? production, along with a down-regulation of oxidant injury. NAC also blocked the up-regulation of TGF-? in in vivo model of VILI. Conclusion TGF-?_1 and TGF-?_2 were up-regulated in VILI. Oxidant injury mediated up-regulation of TGF-? in VILI. NAC, which attenuated oxidant injury and blocked TGF-? up-regulation in VILI, could be a future therapeutic strategy in VILI.

Objective:To observe the clinical efficacy of brain-benefiting and collateral-unblocking needling technique for chronic alcoholic gastritis complicated with depression. Methods:A total of 92 cases with chronic alcoholic gastritis complicated with depression were included in this trial. They were randomly allocated into an observation group (n=46) and a control group (n=46) by random number (envelope) method. Patients in the observation group received the brain-benefiting and collateral-unblocking needling technique. Scalp points included Shenting (GV 24), Xinhui (GV 22), Qianding (GV 23), Baihui (GV 20), Chengguang (BL 6), Tongtian (BL 7) and Luoque (BL 8). Body points included Neiguan (PC 6), Zusanli (ST 36), Zhongwan (CV 12), Gongsun (SP 4), Shenmen (HT 7), Daling (PC 7), Qimen (LR 14), Xinshu (BL 15) and Taichong (LR 3). The control group only received the same body acupuncture as the observation group. The treatment was conducted once a day, 30 min for each treatment, and 10 times made up a course of treatment. The efficacy was observed after 3 courses of treatment, and there was a 2-d interval between two courses. Results:After 3 courses of treatment, the clinical symptoms and gastroscopic features were significantly improved in the observation group than that in the control group. The clinical efficacy, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were significantly better than those in the control group (allP<0.05). Conclusion:The brain-benefiting and collateral-unblocking needling technique can significantly improve clinical symptoms in patients with chronic alcoholic gastritis complicated with depression and substantially alleviate their gastroscopic features, anxiety and depression.

Objective:To observe the clinical efficacy of warm needling moxibustion plus flash cupping for remission-stage peripheral facial paralysis (FP) due to wind-cold.
Methods:Fifty eligible patients were randomized into a warm needling moxibustion group and an acupuncture-cupping group, 25 cases in each group. The warm needling moxibustion group was intervened by acupuncture at Fengchi (GB 20), Yangbai (GB 14) towards Yuyao (EX-HN 4), Xiaguan (ST 7), Dicang (ST 4) towards Jiache (ST 6), Quanliao (SI 18), and Hegu (LI 4), plus warm needling moxibustion at Quanliao (SI 18); the acupuncture-cupping group received flash cupping on the affected side in addition to the intervention given to the warm needling moxibustion group. The two groups were both treated once a day, 10 times as a treatment course, for 3 courses in total. The House-Brackmann (H-B) facial nerve grading system was observed before and after the intervention to evaluate the facial nerve function in the two groups, and the therapeutic efficacies were also compared between the two groups.
Results:The two treatment protocols both can promote the recovery of facial nerve function. The total effective rate was 92.0% in the acupuncture-cupping group versus 72.0% in the warm needling moxibustion group, and the between-group difference was statistically significant (P<0.05).
Conclusion:Warm needling moxibustion plus flash cupping can produce a more significant efficacy than dry warm needling moxibustion in treating remission-stage peripheral FP due to wind-cold.

Objective:To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture. Methods:A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected. Results:Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05,P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05,P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (allP<0.05). Conclusion: EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.