Objective:To study the effect of ?-3 PUFA on the proliferation and apoptosis of tumor cells transfected with transmembrane TNF? (tmTNF?) expression vector containing peroxisome proliferator responsive element (PPRE). Methods:The tmTNF? expression vector was transfected into HL-60 and MCF-7 tumor cells. The effects of ?-3PUFA and/or exogenous tmTNF? on cell proliferation, cell cycle distribution and apoptosis were measured by growth curve,flow cytometry and DNA ladder assay. Results: The production of exogenous tmTNF?gene was located in the cell membrane. After treatment with?-3 PUFA, the expression of exogenous tmTNF? in transfectant was significantly increased. The proliferation was decreased in HL-60 and MCF-7 cells treated with 6.0?10-5 mol/L ?-3 PUFA,but cell cycle was arrested at G0/G1 and the DNA ladder was detectable only in HL-60 cells. These changes were more obvious in transfected cells than non-transfected cells. Conclusion:?-3 PUFA and the product of exogenous tmTNF? gene induced by ?-3 PUFA could inhibit tumor cell growth more effectually,and induction of apoptosis may play a key role in this process.

Objective To study the apoptosis and its molecular mechanism of MCF-7 cells induced by ?-3 polyunsaturated fatty acid(?-3 PUFA)combined with the product of exogenous transmembrane TNF-?(tmTNF-?) gene.Methods The tmTNF-? eukaryotic expression vector containing the PPRE-tk promoter was transfected into the human breast cancer MCF-7 cells.The effects of ?-3 PUFA and/or exogenous tmTNF-? on the cell proliferation and apoptosis were measured by MTT and DNA ladder assay.The activity of caspase-8 was examined by using special fluorescence substrate,and the expression of caspases(1,8 and 9) were analyzed by RT-PCR and Western blotting.The inhibitor of caspases was used to confirm the function of caspase.Results In 6.0?10~(-5)mol/L ?-3 PUFA-treated MCF-7 cells,only the growth suppression was found.In transfected MCF-7 cells after treated with ?-3 PUFA,not only the proliferation capacity was decreased but the DNA ladder was detectable.The expression changes of caspases(1,8 and 9) and caspase-8 activity were obvious in MCF-7 transfected cells treated with ?-3 PUFA.Growth inhibition and apoptosis induced by ?-3 PUFA and tmTNF-? were partly prevented by the special caspase inhibitor.Conclusion These results suggested that up-regulated expression and activity of caspase might promote MCF7 cells apoptosis induced by ?-3 PUFA and exogenous tmTNF-?,indicating that ?-3 PUFA and exogenous tmTNF-? could cooperate in inhibition of the MCF-7 cell growth and induction of apoptosis.caspase network pathway may play a key role in these processes.

Objective:To study the effects of eicosapentaenoic acid (EPA) and retinoic acid (RA) on proliferation and differentiation of HL 60 cells and the corresponding mechanisms. Methods:Flow cytometry was used to assay cell proliferation function, NBT reduction experiments to evaluate cell differentiation, and Western blot to analyse pRB expression. Results:The proliferation index (PI) in all experimental groups were all lower than that of the control , and EPA in combination with RA was the lowest one. NBT reduction experiments showed that the OD value of cells treated by EPA and RA was about 5.9 times of the control, but correspondingly by RA was 2.6 times. Moreover, p21 N ras expression of HL 60 cells were all decreased by treatment with EPA or/and RA. Conclusion:The synergistic effects of EPA and RA on proliferation and differentiation of HL 60 cells may be partly due to the change of p21 N ras expression.l proliferation function,NBT reduc

Objective To construct the transmembrane TNF-? eukaryotic expression vector in n-3 PUFA dependant manner. Methods PPRE-tk sequence was designed and artificially synthesized, and then inserted it into pcDNA3-TNF( ? 1-12)WB plasmid to construct eukaryotic expression vector pPPRE-tk-tmTNF-? by gene recombine techniques. The tmTNF-? protein expression level was observed in MCF-7 transfected cells incubated with EPA by immunofluorescence technique. Results pPPRE-tk-tmTNF-? expression vector was constructed successfully and identified by agarose gel electrophoretic analysis and nucleotide sequence analysis. EPA could increase tmTNF expression levels in time- and dose-dependant manners. Conclusion tmTNF-? expression vector regulated by n-3 PUFA is successfully constructed.

Objective:To investigate the mechanisms of genistein (GEN) affecting the chemosen- sitivity of human breast cancer cell line MDA-MB-453 to paclitaxel (PTX) in vitro. Method:HER2/neu- overexpressing breast cancer cells MDA-MB-453 were treated by GEN, PTX alone or combined in vitro. Cell cycle was measured by flow cytometry. The expression of HER2/neu protein was observed by immunocytochemistry and. Akt, p-Akt, cyclin B1 and CDK1 protein by Western blot. Results:Cell cycle of MDA-MB-453 cells was blocked at G1/S after treatment of GEN, while at G2/M after treatment with PTX alone. Both GEN and PTX did not change the expression of HER2/neu, total Akt and CDK1 in MDA-MB-453 cells, but GEN significantly decreased p-Akt and cyclin B1 level, and PTX obviously increased cyclinB1 level. GEN antagonized the effects of PTX on level of cyclin B1 protein and blockage of G2/M in MDA-MB-453 cells after treatment with GEN and PTX in combination. Conclusion:The antagonism effects of GEN on the increase of cyclin B1 and blockage of G2/M induced by PTX may be one of the mechanisms of GEN affecting the chemosensitivity of MDA-MB-453 cells to PYX.

Objective　To investigate the expression of TGF-β and TGF-β receptor in human breast cancer cell Bcap-37 inhibited by soybean isoflavones. Methods　mRNA and protein of TGF-β1、TGF-βRⅠ in Bcap-37 cells were examined with reverse transcription ploymerase chain reaction(RT-PCR) and immunohistochemistry after cells were treated with daidein or genistein for 1-4 d.The expression of TGF-β1 and TGF-β2 was determined with TGF-β resistance test. Results　The TGF-β1, TGF-β2 and TGF-β recepor increased in Bcap-37 cells at a concentration of 3×10-5 mol/L of genistein. No changes was found when treated with daidzein. Conclusion　Genistein may inhibit the proliferation of Bcap-37 cells and accompany with increasing expression of TGF-β1, TGF-β2 and TGF-β receptor.

Objective To study the relationship between plasma free fatty acids composition and the incidence of nonalcoholic fatty liver disease(NAFLD) .Methods By the design of case‐control study ,105 patients with NAFLD as cases and 110 healthy peo‐ple as controls were enrolled into the study .Plasma free fatty acid levels were determined by gas chromatography .Results High level of plasma palmitic acid(C16 :0)(OR=1 .769) was the risk factors of NAFLD ,while plasma levels of linoleic acid(C18 :2 n‐6) (OR=0 .855) and arachidonic acid(C20 :4 n‐6)(OR=0 .181)were negatively associated with the incidence of NAFLD .Conclusion These findings suggest that a proper ratio of diet fatty acids intake may reduce the risk of NAFLD .

OBJECTIVE To survey the qualification rate of sputum specimen.METHODS The specimen collecting and delivering time(morning,after deep coughing and gargling or not) was investigated.Aerobic bacteria isolated rate was evaluated.RESULTS The mean transportation time in positive aerobic bacteria isolated specimens and negative ones was 75 min and 124 min,respectively.Aerobic bacteria isolated rate was higher in sputum specimen that were microscopically screened for greater than 25 neutrophils,than in sputum specimen that were less then 25 neutrophils and greater than 10 BSE(buccal squamous epithelial) cells per 100? field.CONCLUSIONS Lower respiratory tract specimens should be delivered to the laboratory within 1 hour.Sputum specimen should be collected in the morning and after deep coughing and gargling.Microscopic examination should be mandatory in sputum microbiology,both for specimen evaluation and as a guide to what to look for in culture.

OBJECTIVETo study the effect of combination of eicosapentaenoic acid (EPA) and retinoic acid (RA) on the proliferation and differentiation of HL-60 cells and its mechanisms.

METHODSMTT was used for cell proliferation analysis, NBT reduction experiment for cell differentiation, reverse transcriptase polymerase chain reaction (RT-PCR) for retinoblastoma (RB) mRNA expression, and Western blot for RB protein (PRB) expression.

RESULTSThe proliferation inhibition rates were 35.74%, 24.38% and 42.75% for RA, EPA and combination of EPA and RA. NBT reduction experiments showed that the differentiation induced by EPA and RA was 5.9 times, and by RA was 2.6 times the capacity of the control. The RB mRNA and PRB expression were not changed by EPA, but significantly decreased by the combination of EPA and RA. Moreover, the dephosphorylation rate of PRB was increased by the treatment with EPA or/and RA.

CONCLUSIONThe changes of RB expression and PRB phosphorylation may be one of the mechanisms of the synergistic effects of EPA and RA on the HL-60 cell proliferation and differentiation.

Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Drug Synergism ; Eicosapentaenoic Acid ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; HL-60 Cells ; Humans ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Retinoblastoma Protein ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tretinoin ; pharmacology

Objective To investigate the structure-activity relationship for 21 anthocyanins in inhibiting oxidized injury of endothelial cells,and explore the structural characteristics of anthocyanins closely related to their effects. Methods Endothelial cells were treated by ox-LDL at different concentrations of 50,100,150 or 200 ?g/ml,and MTT assay was used to determine IC50. After pre-incubated for 2 h with different concentrations ( 50,100 or 200 ?mol/L) of anthocyanins and then treated with 100 ?g/ml ox-LDL for another 24 h in endothelial cells,MTT assay was used to detect the cellular viability. After pre-treated for 2 h with different anthocyanins with 100 ?mol/L and treated with ox-LDL for another 24 h,MDA and NO level in the culture media were both measured according to the methods of assay kits. Structure-activity relationship was analyzed according to the respective cellular viability,MDA and NO level. Results Cellular viability was significantly inhibited by ox-LDL in a dose-dependent manner,and the IC50 was 100 ?g/ml. A significant correlation was observed among the effect of anthocyanins on cell viability,MDA production and NO release. The inhibitory effects of anthocyanins in ox-LDL-injured endothelial cells were positively related to the total number of hydroxyl groups and hydroxyl substitutions in B ring. 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring significantly enhanced the inhibitory effect of anthocyanins,yet methoxylation or glycosylation significantly decreased the effect. 6-hydroxylation substitution might attenuate the inhibitory effect of anthocyanins,while substitution at C5 or C5′ showed no significant influence on the effect of anthocyanins. Anthocyanin with monosaccharose substitution was much stronger than that with disaccharose substitution,while there was no significant difference between anthocyanins with glucoside and that with galacotoside substitution. Delphinidin and delphinidin-3-glucoside were respectively the most effectual anthocyanidin or anthocyanin. Conclusion 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring are the main structural requirements for anthocyanins in suppressing ox-LDL-induced injury in endothelial cells.