Objective To investigate the cytotoxic effects of human γδT cells on hepatocellular carcinoma cells in vitro.Methods Normal human PBMC were activated by IPP and rhIL-2.The expansion of γδT cells were analyzed by flow cytometry.The concentration of interferon gamma in the supernatants was detected by ELISA.The cytotoxicity of activated lymphocytes against hepatocellular carcinoma cells were measured by the lactate dehydrogenase release assay in vitro.Results After PBMC activated by IPP and rhIL-2 for 10 days,the proportion of γδT cells were expanded from (4.34±1.79)% to (55.65±6.88 )%.The secretion of interferon gamma expanded significantly after PBMC stimulated by IPP,and reached its peak value at the seventh day.The secretion of interferon gamma is lower if it has no IPP.The activated lymphocytes stimulated by IPP were detected to have cytotoxicity in vitro against human hepatocellular carcinoma ceils.Conclusion The high proportion of γδT cells can be gained in human PBMC activated by IPP,PBMC stimulated by IPP can secret interferon gamma effectively and the activated γδT cells have high cytotoxlcity in vitro against human hepatocellular carcinoma cells.

Objective To construct and express recombinant cecropin B-binding site of luteinizing hormone releasing hormone(CB-LHRH')gene,and to evaluate the anticancer function of CB-LHRH' on human ovarian cancer cell line SKOV3 and human endometrial cancer cell line HEC-1B.Methods The sequence of the cDNA encoding CB-LHRH' was designed,artificially synthesized,verified by DNA sequence analysis and expressed by Bac-to-Bac baculovirus expression system.The expression of CB-LHRH' proteins were identified by western dot blot using rabbit polyclonal antibody against LHRH as the primary antibody.To determine the anticancer effects of the CB-LHRH' protein,ovarian cancer cell line SKOV3 and endometrial adenocarcinoma cell line HEC-1B were treated by different doses of the CB-LHRH' protein.Cell growth inhibition assay was performed using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)5[(phenylamino) carbonyl]-2H-tetrazolium hydroxide(XTT)kit at different times,and cell morphologic changes were observed under the inverted microscope.Results The inhibitory rate of proliferation by CB-LHRH' increased with the increase of dose and time respectively:SKOV3 cell,from(5.03?0.08)% to(53.24 ?1.22)%;HEC-1B cell,from(5.13?0.37)% to(56.16?1.08)%.The inhibitory effect on HEC-1B cell was stronger than that on SKOV3 cell(P

OBJECTIVE: To investigate the cardioprotective effect of exogenous zinc (Zn2+) on the mitochondrial pathway, and to explore its possible mechanism. METHODS: Rat heart tissue-derived H9c2 cardiac cells were cultured, and then randomly divided into control group, ZnCl2 group (1-20 μmol·L-1, 20 min), ZnCl2 plus inhibitor group [PI3K inhibitor LY294002, 10 μmol·L-1and mitochondrial ATP sensitive potassium channel (mKATP) inhibitor 5-HD, 0.5 mmol·L-1, inhibitors treated cells for 10 min and then ZnCl2 20 min] and inhibitor group (10 min). The mitochondrial permeability transition pore (mPTP) opening was evaluated by measuring mitochondrial membrane potential (ΔΨm). Tetramethylrhodamine ethyl ester (TMRE) diacetate fluorescence images were obtained with laser scanning confocal microscopy. GSK-3β and AKT phosphorylation were determined with Western blot. The cells were subjected to simulated ischemia/reperfusion injury, cell viability were determined with flow cytometry. Cells were transfected with constitutively active GSK-3β-S9A (GSK-3β-S9A) plasmid by Fugene 6 transfection kit, mPTP opening was evaluated by confocal microscopy. RESULTS: Compared with the normal, exposure of cells to H2O2 for 20 min caused a marked decrease in TMRE fluorescence, treatment of cells with different dose of Zn2+ prevented the loss of TMRE fluorescence caused by H2O2 with the peak at 10 μmol·L-1. Western blot showed that Zn2+ significantly enhanced the GSK-3β and AKT phosphorylation, the effect that was significantly reversed by LY294002, but not 5-HD. Compared with the normal, ischemia/reperfusion markedly reduced cell viability. Zn2+ applied at ischemia did not increase the cell viability, but significantly increased the cell viability when given at reperfusion. Zn2+ could mimic the specific mPTP inhibitor cyclosporin A (1 μmol·L-1) and prevent the mPTP opening, which was again reversed by LY294002 but not 5-HD. Zn2+ was not able to exert protection in cells transfected with the GSK-30-S9A. CONCLUSION: Zn2+ can induce myocardial mitochondrial protective effect by modulating the mPTP opening through the inactivation of GSK-3β via PI3K/AKT pathway. mKATP may not be involved in the action of Zn2+.

OBJECTIVE: To investigate the cardioprotective effect of astragaloside IV on the mitochondrial pathway, and to explore its possible mechanism.

Chromatography, Micellar Electrokinetic Capillary ; methods ; Liposomes ; Membranes, Artificial ; Pharmaceutical Preparations ; chemistry

OBJECTIVETo investigate the roles of nitric oxide (NO) and eNOS in the pathogenesis of vasovagal syncope (VVS).

METHODSFourteen children with VVS (group A), 10 children with syncope other than vasovagal (group B) and 20 healthy volunteers (group C) were enrolled. Plasma NO levels in groups A and B were determined before and at the termination of the head-up tilt table test (HUT). The G894T polymorphism within the eNOS gene was determined in the three groups.

RESULTSPlasma NO levels in group A increased significantly when syncope attacked from 76.7+/-9.6 micromol/L (before HUT) to 90.0+/-11.4 micromol/L (P<0.05). After the syncope attack was improved, plasma NO level in group A was significantly reduced. There were no statistical differences in plasma NO levels before and after the HUT in group B. Determining the G894T polymorphism within the eNOS gene showed that group A was associated with a higher incidence of the GT gene type as compared to groups B and C (42.9% vs 10%; P<0.05).

CONCLUSIONSPlasma NO may be involved in the pathogenesis of VVS. The increased plasma NO level may be associated with the G894T polymorphism of the eNOS gene.

Child ; Humans ; Nitric Oxide ; blood ; physiology ; Nitric Oxide Synthase Type III ; genetics ; physiology ; Polymorphism, Genetic ; Syncope, Vasovagal ; etiology

OBJECTIVETo compare therapeutic effects between the normal rehabilitation and combined with manipulative method after arthroscopic capsular release for the treatment of severe frozen shoulder, and to evaluate the application value of manipulationp.

METHODSFrom March 2007 to July 2010,arthroscopic capsular release was performed in 48 cases (48 shoulders, 23 left side, 25 right side). All the patients were divided into two groups: control group (11 males and 15 females) and manipulation group (9 males and 13 females). The patients in the control group were treated with conventional rehabilitation procedure, and the patients in the manipulation group were treated with additional manipulation procedure. From the 2nd day after operation, the manipulation was performed for 20 minutes every time, twice daily, and it continued for 10 days. All the cases were followed up and the scale of American Shoulder and Elbow Surgeons Standardized Assessment Form (ASES self-report section) and the range of motion (ROM) were recorded.

RESULTSThe mean follow-up period was (12.54 +/- 5.78) months (ranging from 4 to 25 months). Both ASES scores and ROM in the manipulation group were better than those in the control group at the 1st month after operation, and the difference between the ASES scores and flexion of the shoulder were significant. However, there was no significan difference at the latest follow-up.

CONCLUSIONCompared with the conventional rehabilitative procedure, manipulation following arthroscopic capsular release could promote the process of joint rehabilitation and help the patient back to normal life earlier, but there is no evidence of long term advantage.

Arthroscopy ; Bursitis ; physiopathology ; surgery ; therapy ; Case-Control Studies ; Combined Modality Therapy ; Female ; Humans ; Joint Capsule Release ; Male ; Middle Aged ; Musculoskeletal Manipulations ; Range of Motion, Articular ; Treatment Outcome

OBJECTIVETo verify the regulatory effects of acupuncture on exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) at stable phase.

METHODSThirty cases of COPD were randomly divided into a treatment group (16 cases) and a placebo group (14 cases). Based on specified aerobic exercise, acupuncture was applied in the treatment group and placebo acupuncture was used in the placebo group. The acupoints included Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and so on. The needle did not penetrate into the skin for the placebo group. The treatment was required for 2 to 3 times per week for totally 5 weeks. The indices of exercise tolerance, including 6-min walking distance (6-MWD), exercise time, maximum oxygen uptake (VO2max) forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximum ventilatory volume (MVV), St. George respiratory questionnaire (SGRQ) were observed in two groups before and after treatment.

RESULTS(1) Exercise tolerance: the differences of 6-MWD and exercise time were statistically significant between groups, which were more superior in the treatment group (both P<0.01); the VO2max was significantly increased after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05). (2) Pulmonary ventilation function: the differences of FEV1%, FEV1/FVC and MVV% were statistically significant between groups, which were more superior in the treatment group (P<0.05, P<0.01); (3) SGRQ: the SGRQ was significantly improved after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05).

CONCLUSIONThe acupuncture could improve the exercise tolerance in patients with chronic obstructive pulmonary disease at stable phase, and shorten the onset time of aerobic exercise. Besides, acupuncture combined with aerobic exercise could effectively improve the pulmonary function.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Exercise Tolerance ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

OBJECTIVETo explore the efficacy and feasibility of dog days moxibustion plaster therapy in treatment of allergic rhinitis of different patterns/syndromes.

METHODSAllergic rhinitis of lung deficiency and invasion of cold, spleen qi deficiency and kidney yang deficiency, 56 patients for each pattern/syndrome were randomized into a plaster therapy group and a nasal spray group, 28 cases in each one. In the plaster therapy group, according to the pattern/syndrome differentiation, with literature retrieval method, 3 acupoints of high frequency utility in clinic were selected as one group in acupoint plaster therapy. For lung deficiency and invasion of cold pattern/syndrome, Feishu (BL 13), Fengmen (BL 12) and Hegu (LI 4) were selected. For spleen qi deficiency pattern/syndrome, Pishu (BL 21), Zusanli (ST 36) and Dazhui (GV 14) were selected. For kidney yang deficiency pattern/ syndrome, Shenshu (BL 23), Dingchuan (EX-B 1) and Bailao (EX-HN 15) were selected. Separately, on July 13, 2013, July 23, 2013, August 2, 2013 and August 12, 2013, the aucpoint plaster therapy was applied, 2 to 4 h (1 to 2 h for children) each time. In the nasal spray group, beclometasone dipropionate aqueous nasal spray, 2 presses one nostril each time, 2 to 3 times a day, continuously for 4 weeks. The symptom score and efficacy were compared before and after treatment in the patients of the two groups.

RESULTSThe symptom scores of 3 patterns/syndromes were all apparently improved after treatment as compared with those before treatment in the patients of the two groups (all P<0.05), and the result in the plaster therapy group was better than that of the nasal spray group (P<0.05, P<0.01). For lung deficiency and invasion of cold pattern/syndrome, the total effective rate was 87.3% (20/24) in the plaster therapy group, better than 84.6% (22/26) in the nasal spray group (P<0.05). For spleen qi deficiency pattern/syndrome, the total effective rate was 83.3% (20/24) in the plaster therapy group, obviously better than 76.9% (22/26) in the nasal spray group (P<0.05). For kidney yang deficiency pattern/syndrome, the total effective rate was 79.2% (19/24) in the plaster therapy group, better than 76.9% (22/26) in the nasal spray group (P<0.05).

CONCLUSIONThe dog days moxibustion plaster therapy achieves definite efficacy on allergic rhinitis at the acupoints selected based on the differentiation of different patterns/syndromes and the efficacy is better than beclometasone dipropionate aqueous nasal spray.

Acupuncture Points ; Administration, Cutaneous ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Child ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; therapy ; Yang Deficiency ; drug therapy ; Young Adult

ATP Binding Cassette Subfamily B Member 11 ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; physiology ; ATP-Binding Cassette, Sub-Family B, Member 1 ; physiology ; Animals ; Bile ; metabolism ; Biliary Tract ; physiology ; Drug Interactions ; Drug Resistance, Multiple ; Humans ; Liver ; physiology ; Multidrug Resistance-Associated Proteins ; physiology ; Neoplasm Proteins ; physiology ; Organic Anion Transporters ; physiology ; Organic Anion Transporters, Sodium-Dependent ; physiology ; Organic Cation Transport Proteins ; physiology ; Symporters ; physiology