Objective To investigate the reversal effect of triazole antifungal itraconazole,fluconazole combined with adriamycin on multidrug resistance in leukemia cells.Methods Human chronic myelogenous leukemia adriamycin resistant cell lines K562/ADR cells were incubated with itraconazole,fluconazole,or PSC833 (positive control) combined respectively with adriamycin.CCK-8 assay was used to assess cell proliferation of K562/ADR.The mean fluorescence intensity of intracellular ADR was measured by flow cytometry.The marker of DNA damage γH2AX was detected by Western blot.Results 1 μg/ml itraconazole and 0.5 μg/ml PSC833 can decrease K562/ADR IC50 of adriamycin from 38.30 μg/ml to 8.59 μg/ml and 24.64 μg/ml in a dose-dependent manner.K562/ADR cells were incubated with 1 μg/ml itraconazole or 0.5 μg/ml PSC833 combined respectively with adriamycin for 3 h and 6 h,the mean fluorescence intensity of intracellular ADR were increased 1.54-fold (3 h),1.50-fold (6 h) or 5.97-fold (3 h),5.83-fold (6 h).Itraconazole or PSC833 combined with adriamycin significantly increase the expression of γH2AX in K562/ ADR cells.Conclusion Itraconazole can recover adriamycin sensitivity of K562/ADR by increasing the concentration of intracelullar adriamycin and synergistically increasing DNA damage,but not for fluconazole.

ObjectiveTo explore the role of CXCL16/CXCR6 axis on the metastasis of human lung cancer.MethodsImmunohistochemistry and immunocytochemistry analysis were performed to detect the expression of CXCL16/CXCR6 in human lung cancer samples as well as A549,95D and H292 cell lines,respectively.The effects of CXCL16 on the viability and invasiveness of the three lung cancer cell lines were examined by MTT and in vitro invasion assay,respectively.ResultsHuman native lung cancer cells co-expressed CXCR6 and CXCL16 protein.Compared to the normal lung tissues,there was a stronger specific staining for both CXC16 and CXCR6 protein in the lung cancer tissues.Three kinds of lung cancer cell lines,including A549,95D and H292,all expressed CXCL16 and CXCR6 protein.Furthermore,human recombinant CXCL16 significantly promoted the viability and invasiveness of A549,95D and H292 cells.The stimulated action of CXCL16 on lung cancer cell lines could be effectively blocked by CXCL16 neutralizing antibody.Conclusion CXCL16 and CXCR6 protein are co-expressed in human native lung cancer.CXCL16 is able to promote the viability and invasiveness of lung cancer cell lines,which might be the molecular mechanisms on the metastasis of lung cancer.

BACKGROUND: It is conceivable that bone marrow stem cells can differentiate into smooth muscle cells (SMCs) and contribute to neointimal formation in atherogenesis. However, the mechanism remains unknown. The "milieu-induced-differentiation" hypothesis focuses on the key role of cell-to-cell contact and cytokine on the differentiation of stem cells. Bone marrow mesenchymal stem cells (MSCs) have the potential to differentiate into SMCs.OBJECTIVE: To induce MSCs into SMCs in vitro, and investigate the influence of the differentiated SMCs or cell factors on MSCs differentiation.DESIGN: Controlled experiment in vitro with repeated observation and measurement based on cells.SETTING: Department of Cardiology, First Hospital of Xi'an Jiaotong University.MATERIALS: The experiment was accomplished in the Laboratory of Cardiology, First Hospital of Xi'an Jiaotong University between May 2003 and May 2004. SD rats of either gender were provided by the Animal Center of Xi'an Jiao Tong University, 60-80 g, 90-110 g. The following antibodies were used: Mouse anti human SM-α-actin (NeoMarkers),Mouse anti human Calponin (NeoMarkers), TRITC-coupled goat anti mouse IgG antibody (SBA). Mouse anti rat CD34 conjugated FITC (Santa Cruz), Mouse anti rat CD71 conjugated FITC (Oxford Biotechnology), Mouse anti rat anti-CD90 conjugated PE (Oxford Biotechnology). Lipofectamine 2000 (Invitrogen). PEGFP-N3 (the laboratory).METHODS: Bone marrow mesenchymal stem cells were obtained from rat bone marrow by using percoll density gradient centrifugation. SMCs were isolated by using tissue explantation method. Flow cytometer was used to detect the immunofluorescence stain. Then MSCs and SMCs were identified. MSCs were transfected with pEGFP-N3 by Lipofectamine 2000, while untransfected MSCs were taken as controls. Conditioned culture of MSCs and SMCs: ①MSCs at passage 3 were seeded on chamber slides in a 12-well culture plate. The medium was DMEM containing 0%, 5%,7.5% fetal bovine serum (FBS) and SMCs conditioned medium containing 0%, 5%, 7.5% FBS, respectively. The cells were cultured for 10-14 days and immunofluorescence analysis was performed by using monoclonal antibodies against SM-α-actin, calponin.②Indirect co-culture of MSCs with SMCs were established using a semi-permeable membrane cell culture insert. The inserts were plated into culture well. SMCs were cultured on the inside of inserts while MSCs were added to the outside of inserts, respectively. MSCs were culture alone in medium containing 3%, 7.5% FBS and immunofluorescence analysis was performed by using monoclonal antibodies against SM-α-actin, calponin.③MSCs were transfected with pEGFP-N3. After 24 hours, the MSCs were cocultivated with SMCs at an equal density for 7-14 days.As a control, MSCs were cultured alone. MSCs co-cultured were stained with antibodies against calponin, SM-α-actin. MAIN OUTCOME MEASURES: ①Identification of MSCs by floe cytometer.②cytoplasmic antigen expression of SMCs. RESULTS: ①Immunofluorescence analysis showed that MSCs expressed SM-α-actin, but did not express calponin. As a control, SMCs expressed both SM-α-actin and calponin.②Flow cytometry showed that MSCs expressed CD71 of low level, CD90 of high level and no expression of CD34. ③The MSCs transfected with green fluorescence protein continued to express for 2-3 weeks. ④MSCs grew well in SMCs conditioned medium or different concentrations of FBS. Cell growth was FBS concentration dependent in indirect co-culture system of MSCs and SMCs. Several double-positive cells in direct co-culture system were detected enhanced green fluorescence protein and antibodies against calponin, SM-α-actin. CONCLUSION: ①SMCs conditioned medium and cell factor only promote MSCs growth and cytoplasmic granules increase. But these do not induce MSCs differentiate into SMCs. ②The cell-to-cell contact is essential for MSCs differentiation to SMCs.

Objective To investigate the application of leukoreduction filter in removal of leukocytes in platelet concentrate . Methods Platelet concentrate was prepared by using platelet‐rich plasma method .35 bags of the same type of prepared platelet concentrate were filtered by using leukoreduction filter .The changes of conventional indicators of platelet before and after filter were measured and recorded .The activating platelets indicators PAC‐1 and CD62p were detected by using flow cytometry .The platelet hypotonic shock was measured by biochemical analyzer .The platelet aggregation was measured by using platelet aggrega‐tion instrument .Results After platelet concentrate was filtered by using leukoreduction filter ,leukocyte removal rate was(98 .28 ± 0 .97)% ,platelet recovery rate was(86 .37 ± 2 .84)% .After filtration ,white blood cell count ,platelets ,red blood cells were reduced than before filtration(P<0 .05) .The differences of capacity of platelets ,mean platelet volume and pH before and after filtration were not statistically significant(P> 0 .05) .Before and after filtration ,the expression of platelet activation markers PAC‐1 and CD62p ,platelet aggregation and platelet hypotonic shock were not statistically different(P>0 .05) .Conclusion Platelet leukore‐duction filter could effectively filter leukocytes in platelet concentrate .It would not change the conventional indicators ,and not affect platelet activation ,aggregation and anti‐hypotonic shock capacity significantly .

Objective To evaluate the clinical efficacy and safety of combined therapy of Chinese and West-ern medicine in the treatment of schizophrenia.Methods According to the digital table,130 patients with schizophre-nia were randomly divided into the observation group and control group,with 65 cases in each group.The control group was treated with perphenazine and other Western medicine treatment,the observation group was treated based on the control group with Jieyu Kaiqiao pill added,The clinical curative effect,PANSS score and adverse drug reaction of both groups were compared.Results In the observation group,the total effective rate was 96.92%,significantly high-er than that of the control group (72.31%);After treatment,PANSS score of observation group was (42.68 ±6.11), while it was (55.12 ±6.45)in control group.The PANSS scores of both groups were significantly lower than that before treatment,PANSS score of observation group was significantly lower than that of the control group,there was significant difference between groups(t=6.61,P<0.05);the incidence of adverse reaction of the observation group was 10.77%,significantly lower than the control group (58.46%);the difference was statistically significant (χ2 =32.66,P<0.05).Conclusion Jieyu Kaiqiao pill combined with Western medicine for the treatment of schizophre-nia can significantly improve the clinical efficacy,reduce the incidence of adverse drug reactions.

Objective To investigate the present situation of quality of life in the elderly residents aged more than 60 years in Zhanjiang city and to analyze its influence factors .Methods 418 elderly residents over 60 years sampled by the random sampling in Zhanjiang city were performed the questionnaire survey by using the Chinese Scale of World Health Organization QOL (WHOQOL-BREF) .Results The quality of life in the physical ,psychological and environmental fields in male residents was higher than that in the female residents ;which in the psychological and environmental fields in the high cultural level residents was higher than that in the low cultural level residents ;which in the social relations field in the high income residents was higher than that in the low in-come residents ;there were no statistical differences in the quality of life among the elderly residents with different occupations be-fore retirement and marital status ;the Logistic regression analysis showed that gender and occupation were the influence factors of quality of life self evaluation in elderly residents .Conclusion Gender ,occupation ,educational level and income level are the influ-ence factors of quality of life in elderly residents .

AIM To observe the protective effects of mangiferin on the inflammatory injury and expression of the inflammatory factor in the cerebral tissue of spontaneously hypertensive rats and on MCP-1/CCR2 signal pathway.METHODS Forty spontaneously hypertensive rats were randomly divided into model,benazepril [10 mg/(kg · d)] and mangiferin high,middle and low dose [40,20,10 mg/(kg · d)] groups and other eight rats of same week age served as control group.After consecutive intragastric administration for eight weeks,morphology of the rats' cerebral tissue was observed;their levels of ICAM-1,IL-6 and TNF-α in cerebral tissue were determined by ELISA;their expressions of MCP-1 and CCR2 protein in brain tissue of rats were detected by immunohistochemistry and Western blot and the detection of mRNA expressions of MCP-1 and CCR2 in cerebral tissue of rats were carried out by RT-PCR.RESULTS Compared with the model group,the blood pressure of mangiferin in each dosage group decreased slightly,but there was no significant statistical difference.In the control group and the model group,there was no obvious morphological change in the cerebral tissue.The morphology of rats in the benazepril group,each dose of mangiferin group were all normal.The contents of IL-6,TNF-α,ICAM-1 and MCP1,CCR2 protein and mRNA expression were significantly decreased in the cerebral tissues of spontaneously hypertensive rats.CONCLUSION Mangiferin has obvious anti-inflammatory effects on inflammatory reaction in spontaneously hypertensive rats,its mechanism may be related to inhibiting the expression of MCP/CCR2 signaling pathway.

Objective:To investigate the effect of the leaching liquids of the pure titanium porcelain crowns with barium titanate coating on the proliferation of L929 cells,and to evaluate its cytotoxicity level.Methods:The L929 ceils were cultured in vitro with leaching liquids of titanium porcelain specimens with barium titanate coating (group A),and titanium porcelain specimens(group B) for 1,3 and 5 days respectively.RPMI1640 containing 10％ fetal bovine serum and 1％ phenol was served as the negative(group C) and positive control group(group D),respectively.MTT method was used to test the effects of barium titanate coating on the proliferation of mouse fibroblast L929 cells,the cytotoxicity of the 4 groups was graded.Results:L929 cells of the group showed normal morphology and vigorous growth except group D.During the whole experiment,the absorbance values of group A was greater than that of group C(P ＜0.05).The cytotoxic gradation of group A grade 0.Conclusion:Titanium porcelain specimens with barium titanate coating has a good cytocompatibility.

Objective To test the hypothesis that IL-10 may promoting left ventricular remodeling and cardiac function by modulating extracellular matrix after acute myocardial infarction. Methods Male adult rats were randomly divided into three groups:control group (n=6),MI/AAV2 group (n=16) and MI/AAV2-IL-10 group (n=16). Establishing animal modol of experimental myocardial infarction and recombinant adeno-associated virus type 2 (AAV)/IL-10 (AAV2-rhIL-10) and AAV2 were injected around the ischemic zone. Echocardiography parameters,hemodynamic parameters,left ventricular mass index (LVMI),collagen volume fraction (CVF),perivascular circumferential area (PVCA),collagen type Ⅰ&Ⅲ volume fraction and mRNA levels of collagen type Ⅰ&Ⅲ,matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were compared among the three groups. Results Improved cardiac function was observed in MI/AAV2-IL-10 group shown by echocardiography and hemodynamic examination. Four weeks after myocardial infarction,thickness of different parts of LV was not different in MI/AAV2-IL-10 group and MI/AAV2 group. Nevertheless CVF,PVCA and collagen type Ⅰ volume fraction was significantly descending in remote zone of MI/AAV2-IL-10 group compared with that of MI/AAV2 group. The mRNA expression of collagen type I and MMP-2 was lower in MI/AAV2-IL-10 group than that in MI/AAV2 group. Conclusion Recombinant IL-10 expression mediated by AAV2-rhIL-10 transfection of rats' myocardium promotes LV remodeling and cardiac function after acute myocardial infarction. The promotion was partially achieved by inhibition myocardium collagen deposition.

Objective:To find biomarkers for oral lichen planus by comparing differential expressing proteins. Methods:10 cases of oral lichen planus and normal oral mucosa tissues were collected.Total protein was extracted; differential proteome profiles were established and analyzed by two-dimensional polyacrylamide gel electrophoresis(2D-PAGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Results:(1)The well-resolved,reproducible 2-DE patterns of oral lichen planus and normal oral mucosa were obtained. The results showed that average protein spots were 1 576?67 and 1 608?73 in oral lichen planus and normal oral mucosa respectively, (2) The 13 differential protein spots were identified by Imaging Master 2D image analysis software between oral lichen planus and normal oral mucosa. There were 7 protein spots in oral lichen planus were higher than those in normal oral mucosa, 6 protein spots in oral lichen planus were lower than those in normal oral mucosa. 10 differential expressing proteins were analyzed by mass spectrometry and bioinformation. 4 of them were well characterized including manganese superoxide dismutase (Mn-SOD), Annexin I, vimentin and unknown proteins. Conclusion:Differential expression proteins might be candidate biomarkers for diagnosis of oral lichen planus;and proteomic technique is valuable for screening the diagnostic biomarkers.