Objective: To develop HPLC method for determination of alliin concentration in mice plasma. Methods: The plasma samples were extracted with methanol. The analysis involved a ODS 1 column as stationary phase and distilled water as mobile phase. The flow rate was 0.5mL?min -1 , UV detection wavelength was at 220nm. 5 Fu was used as the internal standard. Results: The calibration curve was linear over a range from 14.8?g?mL -1 to 148.1?g?mL -1 with a correlation coefficient of 0.9992. Conclusion: The method is economic, simple, sensitive and accurate.

Metabolomics is one of the branches of systems biology,which employs nuclear magnetic resonance and mass spectrometry technologies to detect the abnormal metabolites from a variety of body fluids. Study of various molecules and their functions may help to find the disease-related early metabolic marker clusters,understand the molecular mechanism of pathogenesis,which provides broad prospects for the early diagnosis and individualized treatment of malignant tumors.

Objective To explore the treatment effect of human growth factor (NGF) on vascular dementia. Method 47 cases with vascular dementia were randomly divided into two groups (study and control). The study group (24 cases) were treated with NGF, the control group (23 cases) were treated with ordinary medicines.Results The intelligence and cognitive ability in the study group were obviously higher than those in the control group (P<0.01). Conclusion Compared with other methods, NGF has a significant curative effect.

Statement of harmonious medical ethics education is the theoretical attribute in the specific field of medical ethics education in the overall time-value of constructing a harmonious society.This proposition is the inevitable theoretical response to the existent inherent contradictions in medical ethics domain,whose values mainly manifest three dimensions including the value pursuit of a harmonious life,the value construction of the harmony of humanity and utility,the value orientation of harmony of doctors and patients.Moreover,this article discusses the construction of harmonious medical ethics education from three levels,including strengthening the education of care ethics,paying attention to the value of situational ethics,promoting the cultivation of communication ethics.

Objective To investigate the possible mechanism of direct relaxant effects of propofol on tracheal smooth muscle.Methods Nine Japanese long-ear white rabbits of either sex, weighing 2-3 kg were sacrificed by air embolism. The trachea of each rabbit was removed and cut into six strips of (6-8) mm x (12-15) mm. The tracheal strips were suspended in Kreb' s solution, exposed to 95 % O2 and 5 % CO2 mixture and maintained at 37℃. One end of strip was fixed and the other was connected to a tension monitor through a tension transducer. The experiment was divided into six groups: group I propofol; group Ⅱ propofol + indothelin; group Ⅲp Ⅴ propofol + verapamil and group Ⅵ propofol + glibenclamide. In each group the tracheal strips were precontracted with acetylcholine (10-5 mol? L-1). The strips were then exposed to increasing concentrations of propofol (5 ? 10-3 , 10-2 ,2.5 ? 10-2 , 5 ? 10-2 , 10-1 mg?ml-1 ). At each concentration of propofol, the change in muscle tension was allowed to reach the steady state for 30 min. In group I no other drug was added to the Kreb' s solution besides propofol, while in group Ⅱ-Ⅵ before the effects of propofol were determined, the strips were pretreated with indothelin (10-4 mol? ml-1 ) (in group Ⅱ), L-NAME(10-4 mol?ml-1) (in group Ⅲ), methylene blue (10-5 mol?ml-1) (in group Ⅳ), verapamil (10-5 mol?ml-1) (in group Ⅴ) or glibenclamide (10-5 mol?ml-1) (in group Ⅵ). Results The three concentrations of propofol examined (2.5?10-2, 5?10-2 and 10-1 mg?ml-1) produced relaxation of isolated tracheal smooth muscle. In group Ⅱ , Ⅲ , Ⅳand Ⅴ pretreatment of tracheal strips with indothelin, L-NAME, methylene blue or verapamil did not significantly affect the relaxation of tracheal smooth muscle induced by propofol as compared with group Ⅰ. Pretreatment with glibenclamide significantly depressed the direct relaxant effect of propofol on tracheal smooth muscle ( P

Objective To investigate the number of osteoclast (OC) precursor in the peripheral blood of patients with ankylosing spondylitis (AS) and its relationship with serum receptor activator of nuclear factor KB-ligand (RANKL) and Osteoprotegerin (OPG) concentration as well as the disease activity. Methods The peripheral blood mononuclear cells from 8 cases of AS patients and 5 healthy controls were cultured in the medium containing macrophage colony-stimulating factor (M-CSF) (25 ng/ml) and RANKL (40 ng/ml). After being cultured for 14 days, cytochemistry was applied to detect tartrate-resistant acid phosphatase (TRAP) expression and the cells with TRAP expression and ≥3 nuclei were counted and defined as OC. Bone resorption assay was used to demonstrate OC function. ELISA was used to measure serum RANKL and OPG concentration in 23 cases of AS and 17 healthy controls. The relationship was analyzed in AS patients between the number of OC precursors and serum RANKL and OPG concentration as well as the disease activity. The indicators of disease activity were Bath ankylosing spondylitis disease activity index (BASDAI), Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). T test, t' test and Spearman correlation were selec-ted. Results ① Significantly higher OC production was observed in the peripheral blood of AS patients than that of healthy control group. The OC number per ten fields was 10.9±3.4 and 6.2±1.3 respectively (P<0.05); ② There was significant difference between AS patients and healthy controls in serum concentration of OPG and RANKL and the ratio of RANKL/OPG. OPG was significantly higher in AS patients [(157±49) pg/ml] than in healthy controls [(105±20) pg/ml] (P<0.05). RANKL was significantly higher in AS patients [(5.4± 3.8) pg/ml] than in healthy controls [(1.6±0.8) pg/ml] (P<0.05). The ratio of RANKL/OPG was significantly higher in AS patients (0.037±0.026) than in healthy controls (0.016±0.008) (P<0.01 );③Significantly positive correlation was observed between the OC number and the serum concentration of RANKL (r=0.692, P=0.009), the ratio of RANKL/OPG (r=0.813, P=0.001);④ In AS patients, serum concentration of OPG was found to have significantly negative correlation with BASDAI (r=-0.444, P=0.044). Serum RANKL concentration was found to have significantly positive correlation with BASDAI (r=0.543, P=0.011). The ratio of RANKL/OPG was found to have significantly positive correlation with BASDAI (r=0.672, P=0.001). Conclusion ① More OC precursors exist in the peripheral blood of AS patients. These cells may differentiate into osteoclasts, which might play a role in joints destructions in AS;② The mechanism of high OC production is likely to be due to high RANKL concentration which is caused by inflammatory reaction.

0.05. CONCLUSIONS Not premixed bacteria fluid groups don't affect the antibiotic susceptibility test of VITEK2 Compact.

Objective To evaluate the efficacy and safety of combination chemotherapy with vinorelbine and cisplatin in the treatment of metastatic breast cancer resisting to anthracycline and taxane.Methods 20 patients with advanced metastatic breast cancer were given following regimen:Vinorelbine 25mg/m2 was given intraveniously in day 1 and day 8,cisplatin 75mg/m2 was given intraveniously in day 1 or 25mg/m2 was given intraveniously in day1 to day 3,repeated every 3 weeks.Evaluation of response and adverse reactions were practiced every 2 cycles.Results 20 patients were evaluable,among them,2 cases reach CR,8 cases PR,4 cases SD and 6 cases PD,with a median followup of 6 months(4 ～ 18months),16 patients survived and 4 patients died.The median time to progression and the median survival time was 5 months(3 ～ 15 months) and 8 months(4 ～ 18 months) respectively.The treatment well tolerated,The main toxicity was myelosuppression and gastrointestinal reaction with WHO grade Ⅲ～Ⅳ gastrointestinal reaction,neutropena and thrombocytopenia being in 25% 、65% and 10% .Conclusion The regimen of NP is safe and effective in treating advanced metastatic breast cancer resisting to anthracycline and taxane.In addition,it was able to improve survival rate and adverse reactions could be tolerated.

Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.

Objective:To explore the role of γδ T cells in the transdifferentiation of immature dendritic cells(imDC) into osteoclasts(OC). Methods:(1) Peripheral blood mononuclear cells(PBMNC) were cultured with zoledronate(Zol) and recombinant human interleukin-2(IL-2),and PBMNC from healthy volunteers were cultured with granulocyte macrophage colony-stimulating factor (GM-CSF) and recombinant human interleukin-4(IL-4) to differentiate into imDC,which were then cultured with receptor activator nuclear factor к B ligand(RANKL) and macrophage colony-stimulating factor(M-CSF) to differentiate into OC. The purity of γδ T cells,and phenotype changing of OC transdifferentiated from imDC were investigated by flow cytometry. (2) Co-culture system was es-tablished using millicell inserts.γδT cells isolated with immune magnetic bead were placed in the upper compartment and imDC in the lower compartment in the ratio of 10∶1. To explore the role of γδ T cells during differentiation of imDC into OC,tartrate resistant acid phosphatase( TRAP) staining and bone resorption observation staining were used. Tumor necrosis factor-alpha( TNF-α) of supernatant liquid from different cultures was measured using ELISA(Enzyme linked immunosorbent assay) kit. Results:(1) γδT cells can be ex-panded from PBMNC of MM patients, and the production capacity was similar to that of healthy volunteers ( 68. 87%± 20. 94% vs 69. 33%±16. 84%,P>0. 05 ) . ( 2 ) OC could be transdifferentiated from imDC when cultured with RANKL and M-CSF. ( 3 ) The number of TRAP+ multinuclear cell and the absorption area of dentine were significantly lower in the group of imDC indirectly co-cultured with γδ T cells than in the group of control imDC(5.67±0.58 vs 28.33±2.08,4.97%±4.3% vs 28.47%±12.8%, respectively). (4) Under the circumstance of γδ T cell-imDC indirect coculture,TNF-α got significantly higher. Conclusion: γδ T cells might inhibit the transdifferentiation of imDC into OC.γδ T cells-based immunotherapy is expected to be a new treatment for myeloma bone disease.