Objective To study the ER status in Adriamycin-sensitive and Adriamycin-resistant MCF-7 human breast cancer cell lines. MethodsThe status of ER of MCF-7/ADR and parental MCF-7 cells was detected by Western blot. The expression of ER mRNA was detected by RT-PCR .The growth and the sensitivity to Estrogen(E 2) and droloxifene(Dro) of cells were investigated by MTT assay, and the distribution of cell cycle was detected by flow cytometric assay. Results ER and ER mRNA were positive in MCF-7 cells, and negative in MCF-7/ADR cells. In comparison with MCF-7 cells, MCF-7/ADR cells showed lower growth rate, and the cell cycle was arrested at G 0/G 1 phase. E 2 at concentrations between 1?10 -12mol/L to 1?10 -7mol/L significantly stimulated the growth of MCF-7, but did not stimulate the growth of MCF-7/ADR.Dro at concentrations between 10*!?mol/L to 20*!?mol/L significantly inhabited the growth of MCF-7, and the inhibition was dose-dependent. Dro at concentrations below 20*!?mol/L did not inhibit the growth of MCF-7/ADR, dro inhabited the growth of MCF-7/ADR only at the concentration of 20*!?mol/L, and the inhibition was more effective than MCF-7. Conclusions ER was lost in MCF-7/ADR cells,probably at mRNA level. Compared with MCF-7, the growth rate of MCF-7/ADR decreased,MCF-7/ADR cells lost the dependence on estrogen and the sensitivity to endocrine therapy.

Objective To explore the effect of suramin on the epithelial-mesenchymal transition (EMT) and the excretion of transforming growth factor-β1 (TGF-β1) in peritoneal mesothelial cells (PMCs) induced by high concentrations of glucose solution (GS).Methods Cultured PMCs were divided into three groups:(1) normal control group; (2) GS-treated group:cells were treated with 1.5％,2.5％,4.25％ GS for 12 h,24 h,48 h,respectively; (3) Suramin-treated group:PMCs cultured with 4.25％ GS were exposed to different doses of suramin (25,50,100 μmol/L) for 48 h.Expression levels of α-smooth muscle actin (α-SMA) and E-cadherin were detected by Western blotting and the concentration of TGF-β1 in the culture supernatant was determined by ELISA.Results Compared with normal control group,GS-treated PMCs exhibited a time-dependent increase in the expression of α-SMA,and decrease in the expression of E-cadherin.GS also stimulated PMCs to secrete TGF-β1.In the presence of suramin,GS-induced α-SMA expression and TGF-β1 production were reduced,E-cadherin expression was increased.Conclusions Suramin can inhibit high glucose-induced EMT of PMCs by down-regulating the expression of TGF-β1.Suramin may be a novel therapeutic agent for the treatment of peritoneal fibrosis.

Objective To investigate the expression of FHITmRNA and WWOXmRNA in human breast cancer tissues and its relation to clinicopathological and other molecular parameters. Methods With reference to the expression of ?-actin,the expression of FHITmRNA and WWOXmRNA was determined by reverse (transcription)-polymerase chain reaction(RT-PCR) in 51 breast cancer and adjacent breast tissue, and (semi-quantitative) analysis of band densities was performed. The protein expression of estrogen receptor(ER), progesterone receptor (PR), Her-2 gene in the 51 breast cancer lesions was detected by (immunohistochemical) method. Results FHITmRNA and WWOXmRNA expression was significantly different in 54 breast cancer tissue compared to adjacent breast tissue (P0.05); of FHITmRNA and WWOX mRNA was related to axillary lymph node metastasis (P

Objective To investigate whether folate deficiency cause high expression level of interferon gamma (IFN-γ) resulted from IFN-γ gene ( IFNG) hypomethylation and then promote the pathogenesis and development of ulcerative colitis (UC ) in a dextran sulfate sodium (DSS )-induced experimental colitis model in mice .Methods A total of 24 female BALB/c mice were divided into four groups ,six mice in each group , including folate deficient/DSS+ group , standard diet/DSS+ group , standard diet/DSS - group and folate deficient/DSS- group .At the beginning of the sixth week since fed , the mice of model groups were treated with 5% DSS to establish experimental colitis .By the end of the sixth week ,disease activity index (DAI) of colitis and histological changes were evaluated .The folate level of peripheral blood serum of mice were detected by enzyme-linked immunosorbent assay (ELISA ) . The expression of IFN-γ in colonic mucosa of mice was examined by immunohistochemistry . The methylation level of CpG island in the promoter region of IFNG was determined by methylation specific polymerase chain reaction (MSP) .The t test was used for measurement data .Chi square test was performed for comparison between groups of count data . Spearman correlation analysis was used for correlation analysis .Results The folate levels of peripheral blood serum of folate deficiency/DSS+ group and folate deficiency/DSS- group ((2 .70 ± 0 .19) and (2 .80 ± 0 .25)μg/L) were significantly lower than those of standard diet/DSS+ group and standard diet/DSS- group ((13 .62 ± 0 .38 ) and (13 .52 ± 0 .77)μg/L ,t= -63 .33、32 .27 ,both P< 0 .05) ,resepectively .The expression of IFN-γ in colonic mucosa of folate deficiency/DSS+ group and standard diet/DSS+ group were significantly higher than those of folate deficiency/DSS- group and standard diet/DSS- group (χ2 = 22 .18 ,P< 0 .05 ) . And the expression of IFN-γ in colonic mucosa of folate deficiency/DSS+ group was also higher than that of standard diet/DSS+ group (χ2 = 12 .00 ,P< 0 .05) .The expression level of IFN-γ of folate deficiency/DSS+ group and standard diet/DSS+ group was positively correlated with (r=0 .998、0 .953 ,both P<0 .01) .The folate levels of peripheral blood serum of folate deficiency/DSS+ group was negatively correlated with IFN-γexpression level and DAI (r= -0 .880 and -0 .926 ,both P<0 .05) .No abnormal methylation was detected in IFNG promoter CpG island in colonic mucosa tissues of mice of each group . Conclusion In the mice model of DSS induced acute experimental colitis ,folate deficiency may increace the expression of inflammatory factor IFN-γand enhance the inflammation activity of colonic mucosa .

Objective To investigate the effects of Shenfu injection ( SF,a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli) on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heme oxygenase-1 (HO-1). Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups:sham operation group ( Sham group),ischemia reperfusion group ( IR group),and SF Injection group (SF group).The model of focal cerebral ischemia-reperfusion injury was induced by transient occlusion of middle cerebral artery (ischemia for 2 h,and reperfusion for 3,6 h respectively).In SF group,SF ( 10 mg/kg) was intraperitoneally injected duri(n)g reperfusion.Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer.Expression of HO-1 in brain was measured by RT-PCR,while the pathological and ultra structure changes of cerebral tissue were also observed.Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group (P ＜0.01 ),while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group (P ＜ O.01 ).The ultra structure change of brain tissue was less in SF group than that in IR group.Conclusions During early stage of brain IR injury,SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.

Objective:To determine the difference of miRNA expression profiles in 6 types of human cancer cells by microarray technique.Methods:The microarray was prepared,with contained 210 oligonucleotides,including 206 probes complementary with human and mouse miRNA sequences and 4 positive control oligos.MiRNAs were extracted from HeLa(human cervical cancer epithelial cells),MCF-7(human breast cancer cells),A549(human lung adenocarcinoma cells),HT-29(human colonic cancer cells),ES-2(ovarian carcinoma cells),and K562(chronic myelogenous leukemia cells) cells and were labeled with Cy3 for hybridization to the miRNA microarray.The slides were scanned by ScanArrayTMExpress1.0 and images were analyzed by ScanArray3.0 and Cluster3.0;the results were confirmed by Northern blotting and RT-PCR.Results:Totally 115 miRNAs were found to be differentially expressed in the 6 cancer cell lines,with miR-21 expression up-regulated and miR-125b,let-7 expression down-regulated.The expression of miR-17-5p and miR-20a was in cluster and was more higher in ES-2 cells than in other cells.HeLa and MCF-7 cells were located on a single branch of the dendrogram in cluster analysis.Northern blotting showed that both pre-and miR-17-5p expressed in K562,pre-miR-17-5p was weakly expressed in A549 and ES-2 cells,and obvious pre-miRNA expression and weak miRNA expression were noticed in MCF-7,HeLa,and HT-29 cells.RT-PCR showed that expression of pre-miR-17-5p in K562 cells was higher than those in other cells.Expression of miR-21was high in all 6 cell lines and the highest expression was seen in A549 cells.Conclusion:Microarray can be used to detect miRNA expression files in cancer cells,which contributes to the study of the relation between miRNA and tumor.

Objective To investigate the effects of ischemic preconditioning on pneumocyte apoptosis and the expression of HSFT0 after lung isehemia-reperfusion(I/R) in rats and discuss its possible mechanism of extenu-ating ischemia-repedusion injury. Method Thirtysix male Sprague-Dawley rats were randomly divided into three groups [ sham operation(SO ) group, ischemia-teperfusion(L/R) group, and ischemic preconditioning(IP) group],twelve in each group. Lung croas-clamping was used to build the L/R model. In IP group, three cycles of 5-minute-ischemia + 5-minute-reperfusion were given to the pulmonary artery before the procedure. Sham operation rats had a thoracotomy only. Two hours(or five hours) reperfusion was given to both L/R and IP group. Tenninal-deoxynucleotidyl Transferase Mediated d-UTP Nick End Labeiing(TUNEL) was used to evaluate apoptosis. Expression of HSP/0 in lung was observed by immunohistochemical stain and image analysis. Index of quantitative assessment of histologic lung injury(IQA), wet to dry weight ratio(W/D) were measured. The pathological change of lung tissue was observed under both hght and electron microscopy. Statistical analysis was carried out by One-way Anova. Scheffe test was used for intragroup comparison. Results The apoptosis index and expression of HSP70、W/D,IQA of hng tissue in I/R group were higher than those in the sham operation group (P<0.01). Compared with the L/R group, the apoptosis index and expression of HSP70, W/D, IQA of lung tissue significantly decreased (P<0.01), the levels of expression of HSPTO increased significantly in IP group ( P<0.01 ). The pathological and ultrastructure change of lung tissue was better in IP group than those in I/R group. Condusions Ischemic preconditioning can extenuate lung I/R injury by the possible mechanism of increasing the expression of HSPT0 which inhibits the apoptosis during lung I/R injury.

Objective To elucidate the influence of fetal genotype in both non-diabetic gravidas and pregnant women on gestational diabetes mellitus (GDM) through analysis of the genotype discrepancy between maternal and fetal Pro12A1a single nucleotide polymorphism (SNP) of peroxisome proliferator-activated receptor gamma 2 (PPARG2) genes.Methods Pregnant women,who delivered in the Obstetrics and Gynecology Hospital of Fudan University from October 2005 to February 2007,and their newborn babies were selected,and were divided into GDM and control group.The GDM group consisted of 55 gravidas with GDM and 40 newborns born to the GDM mothers,and the control group consisted of 173 healthy gravidas and their 50 neonates.Polymerase chain reaction-denaturing high-performance liquid chromatography was applied to detect the distribution of PPARG2 Pro12Ala alleles in all subjects.The concentrations of plasma fasting blood sugar (FBS) and several bio-markers of lipids,including total cholesterol,triglyceride,apoprotein A,high-density lipoprotein and low-density lipoprotein,were also tested for the mothers.Results (1) No significant difference was found in the frequencies of Pro/Pro genotype between the GDM mothers and control mothers (94.6% vs 90.8%,P > 0.05),nor between the GDM offspring and control offspring (95.0% vs 94.0%,P >0.05) or between the GDM mothers and GDM offspring (P > 0.05).The same was shown in the frequencies of Pro/Ala genotype both between the GDM mothers and control mothers (5.5% vs 9.2%,P >0.05) and between the GDM offspring and control offspring (2.5% vs 3.0%,P > 0.05).(2) Within both GDM and control group,the maternal FBS and various lipids concentrations of Pro/ Pro genotype gravidas showed no significant difference compared to those of Pro/Ala genotype mothers (P > 0.05).(3) Based on the four possible PPARG2 genotype pairs between the mothers and fetuses,Pro/Pro mother and her Pro/Pro fetus,Pro/Ala mother and her Pro/Ala fetus,Pro/Ala mother and her Pro/Pro fetus,and Pro/Pro mother and her Pro/Ala fetus,less Pro/Pro pairs and more Pro/Ala pairs were found in the GDM group than in the control (72.5% vs 92.0%,P=0.014; 27.5% vs 6.0%,P< 0.05).Conclusions Neither the maternal nor the offspring's Pro/Ala genotypes is associated with the genesis of GDM.However,the discrepancy of PPARG2 Prol2Ala polymorphism between mother and her fetus implies a possible cause of GDM.

Objective To evaluate the use of PET/CT in the diagnosis of breast cancer. Methods In this study,33 patients with suspicious breast tumor underwent PET/CT imaging. The images of the breast were analyzed for qualitative assessment of increased tracer uptake and blood perfusion with PET/CT. Results Among 27 cases with pathology proved breast cancer,25 was judged as PET/CT positive,2 was false-negative. Sensitivity, specificity and accuracy of PET/CT in identifying breast cancer were 92.6%,100%,93.9%. Conclusion PET/CT is a reliable and sensitive measure in the diagnosis of breast cancer in vivo.

Objective To study the expression of BRMS1mRNA in human breast cancer tissues and their significance.Methods The expression of BRMS1mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) in 71 breast cancer tissues and adjacent breast tissues,12 patients with benign breast tumors and 12 patients with normal breast tissue,and semi-quantitative analysis of band densities was also performed.Results The expression of BRMS1mRNA in 71 patients with breast cancer and adjacent breast tissue was 0.378?0.046 and 0.918?0.044,respectively;the expression of BRMS1mRNA in 12 patients with benign breast tumors and 12 patients with normal breast tissue was 0.908?0.047 and 0.934?0.028 respectively.BRMS1mRNA expression was significantly lower in breast cancer tissue compared to adjacent breast tissue,benign breast tumors and normal breast tissue(P0.05),but was related to axillary lymph node metastasis and clinical stage(P