Objective To investigate the effect of electroacupuncture at point Zusanli (ST36) on gastric expression of heat shock protein 70 (HSP70) mRNA in rats with stress gastric ulcer and explore the molecular mechanism of its prevention and treatment of gastric ulcer. Method Fifty-one SD rats were randomly allocated to normal control (normal), stress gastric ulcer (model) and electroacupuncture at point Zusanli (electroacupuncture) groups. A model of stress gastric ulcer was made by cold restraint. From the second day after model making, the electroacupuncture group received electroacupuncture at point Zusanli 20 min once daily for three consecutive days, with left and right points Zusanli alternately needled; the model group, the same restraint treatment as in the electroacupuncture group; the normal group, no treatment. Gastric mucosal ulcer index (UI) was calculated using the GUTH’s method. The pathological morphology of gastric tissue was observed by HE staining. Gastric HSP70 mRNA expression was determined by RT-PCR. Result Different numbers and sizes of obvious gastric mucosal ulcers were observed, UI increased significantly (P<0.001) and HSP70 mRNA expression decreased significantly (P<0.05) in the model group compared with the normal group. The pathological change in gastric tissue improved somewhat, gastric mucosal UI decreased significantly (P<0.05) and HSP70 mRNA expression increased significantly (P<0.01) in the electroacupuncture group compared with the model group. Conclusion Electroacupuncture at point Zusanli can upregulate gastric HSP70 mRNA expression and promote the repair of gastric mucosa injuries in rats with stress gastric ulcer.

The Chinese and English published literature regarding the effects of acupuncture on microcirculation in recent years were analyzed, and it was found out that the theory of TCM meridian had subtle connection with microcirculation on structure and function, which could provide a possible breakthrough for the research of meridian essence. Acupuncture, through multiple links, multiple paths and multiple levels, could regulate microcirculation under physiological or pathological condition, which was a bidirectional benign adjustment function, and reflected the characteristic of acupoint specificity. Due to many shortcomings in the current studies, studies with rigorous design, large sample size and multicenter are expected in the further to make a standard systematized research.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Meridians ; Microcirculation

Objective To obtain the recombinant IgE-dependent histamine-releasing factors of Schistosoma japonicum and Clonorchis sinensis (rSjHRF and rCsHRF) and to study the effect of recombinant HRFs to induce histamine release from sensitized rat mast cells. Methods The complete coding regions of SjHRF and CsHRF were cloned separately, and the recombinant plasmids were respectively transformed and expressed in BL21 cells. The soluble recom-binant rSjHRF and rCsHRF were purified. Aliquots of the mast cells obtained from the lungs of OVA-immunized rats were separately incubated with rSjHRF and rCsHRF and the released histamine was measured by the OPT spectrofluorometric procedure. The dose-dependent curves and the kinetics of histamine release induced by rSjHRF and rCsHRF were prepared. Results The recombinant plasmids pET-30-rSjHRF and pET-30-rCsHRF were constructed successfully and the purified soluble recombinant proteins rSjHRF and rCsHRF were obtained by affinity chromatography. rSjHRF and rCsHRF induced histamine release from sensitized mast cells in a dose-dependent manner. At the concentration of 150 mg/L, the average rate of histamine release from sensitized mast cells induced by rSjHRF and rCsHRF were 49.78% and 32.63%, respectively. Histamine release increased with prolonged reaction time and the maximal release occurred at 35min. Conclusion The recombinant parasite-originated IgE-dependent HRFs show an effect of inducing histamine release from sensitized mast cells, suggesting that this protein would play a role in type I hypersensitivity in hosts with parasitic infections.

Aim To investigate the protective effect of ginsenoside Rb1 against apoptosis induced by H_2O_2. Methods H_2O_2 was used to build an oxidative stress-induced injury model in neonatal rat cardiomyocytes. After treated with gensenoside Rb1(20, 40, 80 mg?L -1),the apoptosis rate, the content of malondialdehyde (MDA), and the activity of superoxide dimutase (SOD) of the cardiomyocytes were examined. The intracellular calcium indicated by the fluorescence in cells were measured by the laser confocal microscope. Results Compared with the model group, the apoptosis rate and the content of MDA of the cardiomyocytes decreased greatly (P

Objective:To investigate the relationship between the removal time of 0.2 mm occlusal in-terference and the recovery of masticatory muscle mechanical hyperalgesia in rats.Methods:Forty male Sprague-Dawley rats (200-220 g)were randomly assigned to eight groups,with five rats in each group:(1 )nave group:these rats were anesthetized and their mouths were forced open for about 5 min (the same duration as the other groups),but restorations were not applied;(2 )sham-occlusal interference control group:bands were bonded to the right maxillary first molars which did not interfere with occlu-sion;(3 )occlusal interference group:0.2 mm thick crowns were bonded to the right maxillary first molars;(4)2,3,4,5,and 6 d removal of occlusal interference groups:0.2 mm thick crowns were bonded to the right maxillary first molars and removed on days 2,3,4,5,and 6.The nave group and sham-occlusal interference control group were control groups.The other groups were experimental groups. Bilateral masticatory muscle mechanical withdrawal thresholds were tested on pre-application days 1 ,2, and 3,and on post-application days 1 ,3,5,7,1 0,1 4,21 and 28.The rats were weighed on pre-application day 1 and on post-application days 1 ,2,3,4,5,6,and 7.Results:Between the nave group and the sham-occlusal interference control group,there was no significant difference in the mastica-tory muscle mechanical withdrawal threshold of bilateral temporalis and masseters at each time point.No significant difference was detected between the contralateral side and ipsilateral side in experimental groups (P>0.05 ).In the 2,3,4,and 5 d removal of occlusal interference groups,the masticatory muscle mechanical withdrawal thresholds decreased after occlusal interference and increased after removal of the crowns and recovered to the baseline on days 7,1 0,1 4,and 1 4,respectively [the masticatory muscle mechanical withdrawal thresholds of right masseter muscle were (1 37.46 ±2.08)g,(1 39.02 ± 2.1 1 )g,(1 40.40 ±0.98)g,(1 38.95 ±0.98)g,respectively].In the 6 d removal of occlusal inter-ference group,the masticatory muscle mechanical withdrawal threshold increased after removal of the crowns and became stable since day 1 4.There was a significant difference between the 6 d removal of oc-clusal interference group and the sham-occlusal interference group on day 28(P<0.05),the masticatory muscle mechanical withdrawal thresholds of right masseter muscle were (1 31 .24 ±0.76 ) g and (1 41 .34 ±1 .43)g,respectively.Conclusion:After removal of the 0.2 mm thick crown within 5 days, the mechanical hyperalgesia of the rats could reverse completely.The mechanical hyperalgesia of the rats could only be relieved,but not reverse completely after removal of the 0.2 mm thick crown on day 6.As the time went on,even minor occlusal interference could cause irreversible mechanical hyperalgesia of masticatory muscles.This study suggested that occlusal interference caused by dental treatment should be eliminated as soon as possible,to avoid irreversible orofacial pain.

Objective Androgen receptor ( AR) is extensively expressed in breast cancer and influences the proliferation of the malignant cells.Our study aimed to investigate the effect of AR on estrogen receptor (ER)-positive breast cancer. Methods ER-positive MCF-7 breast cells were exposed to various concentrations of agonist dihydrotestosterone ( DHT) or antagonist bicalutamide or left untreated .Then the proliferation and apoptosis of the cells were determined by MTT assay , cell counting , and flow cytometry , and the expressions of the proteins related to cell cycle regulation were detected by Western blot . Results The relative gray value of AR was significantly increased in the DHT group (1.055 ±0.020) but decreased in the bicalutamide group (0.705 ±0.010) as com-pared with the blank control (0.795 ±0.020) (both P<0.05).Flow cytometry showed that the early apoptosis rate of the breast cancer cells was markedly higher in the DHT group ([51.20 ±0.312]%) but lower in the bicalutamide group ([2.410 ±0.367]%) than in the blank control ([3.540 ±0.375]%) (both P<0 .01). In comparison with the control group , the expressions of the p53, p73 and p21 proteins in the MCF-7 cells were remarkably up-regulated in the DHT group but down-regulated in the bicalutamide group ( both P<0.05). Conclusion AR inhibits the proliferation of ER-posi-tive breast cancer cells , which suggests that it may be a potential ther-apeutic target for ER-positive breast cancer .

[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .

Objective Tumor suppressor gene p53 can inhibit tumor cell growth, arrest cell cycle, and promote apoptosis.Howev-er, the effects of p53 on the pathogenesis of breast cancer have not been fully elucidated.The aim of this study was to explore the expression of p53 protein and the correlation with clinical pathologic features in breast cancer.Furthermore, the regulatory effects of 5-aza-2′-deoxycyti-dine on p53 in breast cancer cell line were also studied. Methods The expression of p53 protein in 80 cases of breast cancer and normal and adjacent tissue were determined by the immunohistochemical staining .The expressions of p53 mRNA and p53 protein in breast cancer cell line were determined by RT-PCR and Western blotting. Results The positive rate of p53 in breast cancer (41.25%) was higher than that in the normal and adjacent tissue (22.5%) (P<0.01).The expression of p53 was not significantly correlated with age, grade, stage and lymph node metastasis (P>0.05).The low expression of p53 both in mRNA and in protein levels were found in breast cancer cell line of MCF-7.The expres-sion of p53 increased after 5-aza-2′-deoxycytidine administration . Conclusion p53 is highly expressed in breast cancer , which may play an im-portant role in the development and progression of breast cancer. 5-aza-2′-deoxycytidine, up-regulating the p53 expression in breast cancer cell line, which provides the evidents for the development of therapeutic drugs for the patients with low expression of p53 breast cancer.

Objective:To examine the expression ofphospholipase A2 receptor (PLA2R) in renal tissues and the level of anti-PLA2R antibody in serum in patients with idiopathic membranous nephropathy (IMN) and secondary membranous nephropathy (SMN),and to evaluate their diagnostic value in IMN.Methods:A total of 73 patients,who were diagnosed between May,2014 and February,2015 in the Department of Nephrology of the Second Xiangya Hospital,Central South University,were divided into three groups:an IMN group (n=48),an SMN group (n=17) and a minimal change disease group (n=8) according to the renal biopsy.PLA2R expression in renal tissues and the level of antiPLA2R antibody in serum were detected by indirect immunofluorescence technique.Results:The positive rate and fluorescence intensity for PLA2R in the renal tissues in the IMN group were higher than those in the SMN group (91.7％ in the IMN group vs 29.4％ in the SMN group,P＜0.05),while the positive rate and serum level for anti-PLA2R antibody in the IMN group were higher than those in the SMN group (85.4％ in the IMN group vs 29.4％ in the SMN group,P＜0.05);the expression of PLA2R in renal tissues and the serum level for anti-PLA2R antibody were not detected in the minimal change disease group,The serum level of anti-PLA2R antibody was positively correlated with 24 h urine protein (r=0.432,P＜0.01) and negatively correlated with serum albumin (r=-0.307,P＜0.05).Conclusion:The expression of PLA2R in renal tissues and the serum level of anti-PLA2R antibody might be potential markers for diagnosis oflMN.

ObjectiveTo evaluate the clinical and radiological efficacy of TNFR Ⅱ -Fc combined with methotrexate( MTX ) in treatment of patients with moderate and severe rheumatoid arthritis.MethodsThree hundred and ninty-six RA patients were randomized into the combined treatment group,the TNFR Ⅱ -Fc only group and MTX only group.All patients were treated for 24 weeks.ACR-N,ACR20,ACR50,ACR70,DAS28-ESR and Sharp score of both hands were measured for efficacy,and the side-effects were analyzed by one-way ANOVA.Results After 24-week therapy,the ACR-N of the combined treatment group [( 12.79±9.24)％-year] was significantly improved than that of the TNFR Ⅱ-Fc only group [(9.56±11.16)％-year,P＜0.05] and that of the MTX only group[(5.08±11.10)％-year,P＜0.05],and the TNFR Ⅱ-Fc group was significantly improved than that of the MTX group(P＜0.05).The ACR20 response rate of the combined group(80.4％) was significantly higher than that of the TNFR Ⅱ -Fc group(71.1％,P＜0.05) and the MTX group(56.7％,P＜0.01 ).The ACRS0 response rate of the combined group(53.6％) was significantly higher than that of the MTX group(30.8％,P＜0.01 ).The ACR70 response rate of the combined group was 27.7％,which was significantly different from that of the TNFR Ⅱ -Fc group (15.8％) and MTX group (7.7％,P＜0.05or P＜0.01 ).DAS28-ESR in the combination group was significantly reduced than those of the TNFR Ⅱ -Fc group and MTX group,and the DAS28-ESR of the TNFR Ⅱ -Fc group was significantly reduced than MTX group.The average total Sharp score of both hands,which demonstrated the radiographic changes,was significantly reduced in the combination group than the MTX group(P=0.03).The total adverse events in the combined group(40.9％) was significantly high than that of the MTX group(28.8％,P＜0.05).Conclusion TNFR Ⅱ -Fc combined with MTX can effectively control the activity of RA and radiological progress.