Objective To observe the effect of atropine and N-nitro-L-arginine methyl ester (L-NAME) on the pressure of lower esophageal sphincter (LESP) regulated by electro-acupuncturing (EA) at Zusanli acupoint (He-Sea, st 36) of stomach meridian and explore the neural mechanism of EA. Methods Forty-eight rats were divided into six groups:control group, EA group, atropine group, atropine+EA group, L-NAME group and L-NAME+EA group. LESP was observed and recorded by using three-channel perfusion manometric measurement system. Results LESP increased significantly under or after EA at Zusanli acupoint. Cholinergic M receptor blocker partly abolished the influence of EA on LESP, but EA could restore the decreased pressure of cholinergic M receptor blocked rats. Nitric oxide synthase (NOS) inhibitor increased LESP, and EA could make it higher. Conclusions The main efferent pathway of regulating effect of EA at Zusanli acupoint on LESP is via the chlinergic nerve of vagus, and other mechanism possibly exits.

Objective To study the expression of miR-320d in diffuse large B cell lymphoma (DLBCL),and its correlation with prognosis of DLBCL.Methods Sixty cases of DLBCL with the follow-up data were collected from Shanxi Cancer Hospital,and were examined by immunohistochemical EnVision method for CD3,CD10,CD20,bcl-6 and Mum-1.The DLBCL were classified into germinal center B cell-like (GCB) and non-germinal center B cell-like (non-GCB) subtypes according to Hans' algorithm.Agilent Human miRNA Microarray 16.0 was used to select the miRNAs on 24 cases paraffin-embedded tissue of DLBCL.The expression levels of miR-320d in 62 cases were examined by TaqMan real-time polymerase chain reaction.Eleven cases of reactive hyperplasia of lymph node were selected as control.Results In 62 cases of DLBCL,22 cases (35.5 ％) were GCB and 40 cases (64.5 ％) were non-GCB subtypes.The expression of miR-320d in GCB was 3.43 times as much as non-GCB subtypes (P =0.034),and in reactive hyperplasia of lymph node it was 5.65 times as much as in DLBCL (P ＜ 0.001).The low expression groups of miR-320d was significantly correlated with shorter overall survival (P =0.021).Multivariate Cox proportional hazard regression analysis (including the IPI scores) revealed that the down-regulated miR-320d was the independent predictor in DLBCL (RR =2.434,95 ％ CI 1.148-5.159,P =0.020).Conclusions The down-regulated expression of miR-320d might be considered as a distinct subgroup with poor prognosis.

Objective At present study,we sought to further define the significance of endoscopic and pathological findings in the diagnosis of acute intestinal GVHD.Methods The clinical、endoscopic and pathological data in 23 patients with proven acute intestinal GVHD were retrospectively reviewed.Results The endoscopic features of the ileum and colon varied from being grossly normal、loss of vascular markings、diffuse mucosal erythema、edema and friability to erosion、ulceration、mucosal sloughing and bleeding.Pathological findings included crypt epithelial cell apoptosis and dropout、crypt destruction and variable lymphocytic infiltration of epithelium and lamina propria.Conclusion Endoscopic and pathological evaluation of the ileum and colon can be used to diagnose acute intestinal GVHD.Endoscopy may play a significant role in establishing early diagnosis for acute intestinal GVHD in patients following allogeneic BMT,but pathological evaluation of the ileal and colonic mucosa is needed to confirm the final diagnosis.

Objective To study the function of the toll-like receptor-4 (TLR-4) signaling pathway in the synthesis and secretion of pulmonary artery smooth muscle cells of rats with COPD. Methods The primary pulmonary artery smooth muscle cells (PASMCs) of rats with COPD were digested, separated and purified. Then they were randomly divided into control group, LPS group, TLR4 inhibitor group (TAK242) and LPS + TLR4 inhibitor group. RT-PCR, Western blot were used to detect the expression level of TLR-4 and NF-κB among groups. The levels of IFN-γ and PDGF-AB in supernatant with PASMCs in each group were detected by ELISA. Results LPS increased the expression of TLR-4、 NF-κB and the levels of IFN-γ and PDGF-AB. The expression of TLR4, NF-κB and the levels of IFN-γ and PDGF-AB were significantly reduced after inhibiting the expression of TLR4(P < 0.05). Conclusion TLR-4 signaling pathway involved in the inflammatory response and pulmonary vascular remodeling which increased the synthesis and secretion of IFN-γ and PDGF-AB in PASMCs. It provides a theoretical approach for the early intervention of clinical with COPD.

Objective To investigate the clinical manifestations,endoscopy and pathological features of Behcet's disease (BD) in gastrointestinal systems in elderly patients.Methods Totally 8 patients aged ≥ 60 years with BD were collected.Clinical characteristics and endoscopic performances of gastrointestinal tracts in patients were summarized.All patients were received gastrointestinal biopsy and the histology change was observed.The vasculitis of gastrointestinal tracts was confirmed by histochemical staining.Results The proportion of patients with BD involving the esophagus was 16.7％ (9/54) and among whom 6 cases (11.1 ％) were elderly,so the proportion of BD patients with esophageal lesions in elderly patients was 66.7 ％ (6/9).The proportion of patients with BD involving the ileocolon was 9.3％ (5/54) and 4 cases (7.4％) were elderly patients,so the proportion of BD patients with ileocolon lesions in elderly patients was 80.0％ (4/5).6 patients had retrosternal chest pain,among whom 2 patients with dysphagia and 4 patients with abdominal pain and hematochezia.In the laboratory tests,the levels of erythrocyte sedimentation rate (ESR) and C reactive protein were elevated in 4 cases,and antinuclear antibodies (ANA) and anti-neutrophil cytoplasm antibody(ANCA)were negative in all patients.In the physical examination,oral and genital ulcers in 6 patients,and a history of recurrent oral and genital ulcers were found in the other 2 patients.6 patients had ophthalmia and 4 patients with skin lesions.6 patients had multiple esophageal ulcers,in which shallow ulcers was more common,and the surface of ulcer was clean and white with congestion and edema at the periphery of ulcers.BD involving the ileocecal and ascending colon were found in 4 cases.Typical small vessel vasculitis was the pathological performance.The retrosternal chest pain disappeared and the esophageal ulcers were healed after the treatment.Conclusions Gastrointestinal involvement in BD patients is associated with the the course of disease.Longer duration can increase the possibility of gastrointestinal involvement in BD.Elderly BD patients are more common with esophageal involvement.Rctrosternal chest pain may be a relatively specific clinical symptom in BD involving esophageal tract.Glucocorticoids and (or) thalidomide are effective for the treatment of BD with esophageal involvement.

Objective To study the influence of Rhizoma typhonii extract on the growth of glioma SHG-44 cells cultured invitro,and to explore the mechanism of the inhibitory effect of Rhizoma typhonii extract on the growth of glioma cells.Methods The SHG-44 cells were cultured and divided into blank control group and 8, 40, 200, 1 000μg·L-1 Rhizoma typhonii extract groups.The inhibitory effect of Rhizoma typhonii extract on the growth of glioma SHG-44 cells was measured by MTT assay. The secretion levels of Bax and caspase-3 proteins were examined by ELISA assay. The expression level of caspase-3 protein was examined by Western blotting method. Results Compared with blank control group, the inhibitory rates of the growth of SHG-44 cells in 200, 1 000μg·L-1 Rhizoma typhonii extract groups at 24 h,and 8,40,200,and 1 000μg·L-1 Rhizoma typhonii extract groups at 48 h were significantly increased (P<0.05 or P<0.01).The secretion levels of Bax and caspase-3 proteins in 40,200,and 1 000μg· L-1 Rhizoma typhonii extract groups at 48 h were increased compared with blank control group (P<0.05 or P<0.01 ). Compared with blank control group, the expression levels of caspase-3 protein in different doses of Rhizoma typhonii extract groups were increased significantly (P<0.01). Conclusion Rhizoma Typhonii extract can inhibit the growth of cells through up-regulating the expression of Bax protein,increasing the expression level of caspase-3 protein and activating apoptosis pathway.

BACKGROUND:Recently,application of stem cells and growth factor to promoting lung regeneration in repair of emphysema lesion has been a hot focus in study.Thus,it is worth to pay attention on whether stem cells carrying relevant foreign growth factor gene can repair emphysema lesion.OBJECTIVE:To evaluate the effidency of adenovirus vector mediated green fluorescence protein(Ad-GFP)transfecting bone marrow mesenchymal stem cells(BMSCs)and its effect on the cell proliferation,to explore oriented migration of intravenously administrated BMSCs transfected with Ad-GFP in the lung tissues of pulmonary emphysema rats.METHODS:MSCs were separated and purified from the bone marrow of rats by density gradient centrifugation and by adherence.At different multiplicity of infection(MOI),transfection efficiency was observed by laser confocal microscopy.At 48 hours of transfection,MTT method was used to evaluate the proliferation of MSCs.A total of 16 Wistar rats were randomly divided into emphysema model group and control group(n=8).Model rats were established by exposure to cigarette smoke.MSCs,transfected with Ad-GFP,were grafted into the body of rats via tail vein.Lungs derived at 24 hours after implantation,and frozen sections were made.Migration and survival of MSCs in the lung tissues were observed by fluorescence microscopy.RESULTS AND CONCLUSION:MSCs from Wistar rats were successfully cultured,grew well and infected by Ad-GFP.The highest transfection effincincy(88.42 %)could be achieved at MOI of 200.Green fluorescent protein labeling had little effect on proliferation of MSCs by different MOI(P＞0.05).At 24 hours posttransplantation,the green fluorescence-positive tissue was Found in the lung tissues of emphysema model group and control group.Compared with control group,the expression of GFP in lung tissues was higher in emphysema model group(P＜0.05).These suggested that introduction of target gene cannot affect proliferation and homing property of BMSCs.

Objective To investigate the protective effects of folic acid on the oxidative damage that ox-LDL (oxi?dized low-density lipoprotein receptor 1) render to human umbilical vein endothelial cells (HUVEC). Methods HUVECs were injured by ox-LDL (120 mg/L) for 24 h while they were incubated with various concentration of folic acid (0,15, 60, 150, 225, 300, 375 nmol/L). Then HUVECs were cultured in media contains same concentration of folic acid but without ox-LDL for 72 hours. Finally, HUVECs were harvested after 24, 48, 72 and 96 h. The morphological changes were observed us?ing inverted microscope and cell viability were examined by MTT. Results Various concentrations of folic acid (0,15, 50, 100, 200 and 500 nmol/L) has no obvious promotion or inhibition effect in growth of normal HUVEC (P>0.05). However, compared with the ox-FA-def group, 150, 225, 300 and 375 nmol/L of folic acid promoted proliferation of HUVECs with 96 and 120 hours of incubations (P < 0.05). Folic acid of 60, 150, 225, 300 and 375 nmol/L promoted the proliferation of HUVECs with 72 h and 96 hours of incubation (P<0.05). Conclusion High dose folic acid can reduce the ox-LDL oxida?tive damage on HUVEC in a concentration dependent manner.

Objective:To investigate the induction effect of NPPB,a chloride channel blocker,on the apoptosis of human glioma SHG-44 cells,and to explore its mechanism. Methods:The SHG-44 cells were cultured in vitro and divided into control group and NPPB groups (50,100,200 μmol· L-1 ).The cell viability was detected by MTT assay.The apoptotic rates were detected by flow cytometry.The expression levels of Bax, Bcl-2 and caspase-3 were detected by immunohistochemical analysis and Western blotting method.Results:Compared with control group,the cell viabilities of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups after treated for 24 and 48 h were decreased significantly (P < 0.01).The results of flow cytometry showed that the apoptotic rates of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups were 24.64% and 41.85%,and they were higher than that in control group (4.17%) (P <0. 01).The immunohistochemical analysis and Western blotting results showed that the expression levels of caspase-3 and Bax proteins in SHG-44 cells in 100 μmol · L-1 NPPB group were increased (P < 0.05 or P < 0. 01 ), and the expression level of Bcl-2 protein was decreased (P < 0.05 ). Conclusion:NPPB could induce the apoptosis of human glioma SHG-44 cells by the down-regulation of the expression of Bcl-2 and the up-regulation of the expression of Bax,and the activation of caspase-3.

AIM:To explore the effect of component II of broccoli polypeptide on the apoptosis in glioma cells . METHODS:Human glioma SHG-44 cells were cultured and divided into control group and 3, 10, 30 and 100 mg/L com-ponent II of broccoli polypeptide groups .Cell viability was detected by MTT assay .The apoptotic rates were examined by Annexin V/PI staining.The morphological changes of the cells were observed under inverted microscope .The protein ex-pression of Bax and Bcl-2 was detected by immunocytochemistry and Western blotting .The protein level of caspase-3 was also examined by Western blotting .RESULTS:Treatment with component II of broccoli polypeptide for 24 h, 48 h or 72 h induced significant inhibition of viability of SHG-44 cells in a time-and dose-dependent manner .The results of Annexin V/PI staining showed that the apoptotic rates were increased in treatment groups in a dose -dependent manner .The density of glioma cells was decreased after treated with increasing concentrations of the drug , and the apoptotic bodies were ob-served under inverted microscope at 72 h.The results of immunocytochemistry and Western blotting showed that the expres-sion of Bax protein was increased but Bcl-2 protein expression was decreased , and the ratio of Bax/Bcl-2 was increased sig-nificantly compared with control group (P<0.05 or P<0.01).The level of caspase-3 protein was increased in 30 and 100 mg/L component II of broccoli polypeptide groups compared with control group (P<0.01).CONCLUSION:The compo-nent II of broccoli polypeptide increases the ratio of Bax /Bcl-2 and activates caspase-3 protein, thus inducing the apoptosis of glioma cells.