To clarify the molecular mechanism of SAHA in the cell proliferation of ER-positive breast cancer cell line MCF-7 induced by leptin. Methods Human breast cancer cell MCF-7 was incubated with SAHA and/or leptin, and cell viability, apoptosis and cell cy-cle of MCF-7 cells were detected by Muse Cell Analy-zer. The expression of proteins related with apoptosis was determined by apotosis antibody array. Results Real-time cell proliferation assays indicated that the in-duction effect of leptin for MCF-7 cells reached the peak at a concentration of 0. 625 nmol · L-1 . SAHA reduced the viability of MCF-7 cells, induced G0/G1 phase arrest in the cell cycle, and triggered the apopto-sis. Meanwhile, SAHA significantly induced the pro-tein expressions of some apoptotic factors, including Bax, Caspase-3, TRAIL DR5, p21CIP1, and inhibited the expressions of Claspin, Clusterin, x-linked inhibi-tor of apoptosis protein(XIAP) and survivin. Howev-er, leptin had reverse effects on the related expression of the proteins. Conclusion The effects of cell prolif-eration by leptin and SAHA treatment in breast cancer ER positive cell line MCF-7 may involve in the activa-tion of apoptosis pathway, in particular with releasing of Caspase-3 trigged by endogenous mitochondrial ap-optosis pathway.

Aim To investigate the specific binding sites that histone deacetylases 1(HDAC1) and estrogen receptor α(ERα)can be recruited to regulate the transcriptional activity of p21WAF1/CIP1 promoter in the breast cancer MCF-7 cells, and to clarify the molecular mechanism of suberoylanilide hydroxamic acid(SAHA) and leptin regulating p21WAF1/CIP1 promoter function.Methods The breast cancer MCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours, and treated with 20 μmol·L-1 SAHA(SAHA group) or 0.625 nmol·L-1 leptin(Leptin group) for 24 hours.The cells that were cultured in complete RPMI 1640 medium without any treatment were assigned as control group(Basal group).The cell lysis was prepared and incubated respectively with anti-HDAC1 and anti-ERα antibody by chromatin-immunoprecipitation(ChIP) method overnight at 4℃.The DNA-ChIP was followed the manufacturer′s protocol for the assay.DNA fragments binding anti-HDAC1 and anti-ERα antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21WAF1/CIP1 promoter region(+2~-4 000 bp) binding with antibody was detected by real-time PCR and analyzed by 2-△△CT method.Results In basal group, HDAC1 and ERα had high affinity with the f1 and f8 fragments of p21WAF1/CIP1 promoter compared to the f4 fragment.In SAHA group, the binding ability of HDAC1 and ERα to the f1 and f8 fragments of p21WAF1/CIP1 promoter was significantly lower than that of the control, while reversing to reach the peak after leptin treatment.Conclusions HDAC1 and ERα can be recruited to p21WAF1/CIP1 promoter by the cell proliferation signal during the proliferation of breast cancer MCF-7 cells.The DNA f1(from 0 to-400 bp) and f8(from-2 800 to-3 200 bp) fragment in the upstream of p21WAF1/CIP1 promoter are the target functional region for the binding with HDAC1and ERα.

Objective To explore whether the antagonistic effect of decorin (DCN)on progression of glomeruloselerosis is associated with the inhibition of the expression of type I and type II transforming growth factor-? receptors (TGF-?R I and TGF-?R II )in mesangial cells (MsC). Methods RT-RCR and Western blot analysis were used to detect the expression of TGF-?R I and TGF-?R II mRNA and their proteins on cultured rat MsC stimulated by exogenous TGF-?1. Lipofectin-mediated method was used to transfect DCN vector into MsC. After screening and identifying of transfected MsC, RT-PCR and Western blot analysis were adapted to detect the changes of TGF-?R I and TGF-?R II expression respectively. Results The expression of TGF-?R I and TGF-?R II mRNA and their proteins on normal MsC stimulated by exogenous TGF-?1 increased in time-dependent manner and reached the peak at 24th hour. Compared with normal and untransfected MsC (1P-1), the mRNA and protein expression of TGF-?R I and TGF-?R II on MsC (3D-5, 7D-1) transfecled by DCN gene decreased significantly, and DCN gene transfection could antagonize the increase of mRNA and protein expression of both receptors caused by exogenous TGF-?1. Conclusions The expression of both TGF-?R I and TGF-?R II decreases obviously in MsC overexpressing DCN gene, which may be one of the importan antagonistic mechanisms of decorin involved in the development of glomeruloselerosis mediated by TGF-?.

Objective To explore the inhibitory effect of decorin(DCN) on rat mesangial cell (MsC) growth and on the expression of mitogen-activated protein kinases (MAPKs) and p21 protein. Methods Lipofectin-mediated method was used to transfect DCN vector into MsC. After the screen and identification of transfected MsC, DCN-containing supernatant was collected and added into the culture medium of normal MsC, then flow cytometer was used to detect the cell cycle. Western blot analysis was used to explore the changes of MAPK and p21 protein. Immunofluorescence was adapted to detect the expression of p21 on cultured MsC. Results Compared with normal MsC, the number of G2-M cells treated with DCN-containing supernatant decreased to 35%(P

ObjectiveTo induce human peripheral blood mononuclear cells differentiate into hepatocyte-like cells by hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro and determine whether PKH26 could be used to serve as an effective tracer for the cells,and observe the ability of transplanted hepatocyte-like cells differentiate into hepatic cells in nude mice.MethodsGroup A and B were set up respectively.In Group A,mononuclear cells were cultivated without hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in cell culture.They were used as negative control group.In Group B,mononuclear cells were cultured with the administration of both HGF and FGF-4 to induce the differentiation into liver hepatocyte-like cells.The changes in cell morphology were observed and the expressions of AFP and CK 19 were detected by immunocytochemical staining in two groups at different times after induction.The hepatocyte-like cells differentiated from human peripheral blood mononuclear cells labeled by the fluorescent dye PKH26 injected into caudal vein in nude mice is experimental group.The nude mice injected with equal amount of normal saline in control group.The migration of the labeled cells into the liver are observed by the fluorescence microscope in the hepatic tissue sections of nude mice and the expressions of ALB were detected by immunocytochemical staining two weeks after the cells transplantation.ResultsCells in group B have a strong proliferative activity.It becomes large and oval,grows in colonies following induction.Cells in group A that showed spherical shape when peripheral blood mononuclear cells were just isolated are gradually becoming inconformity in morphology,spindle or fibroid,and a few cells are round:cells developed apoptosis and cracked following incubation.The expressions of AFP and CK19 were positive after induction in group B as detected by immunocytochemicat staining.Inversely,the expressions of AFP and CK19 were negative in group A after incubation.The experimental group showed numerous PKH26 labeled cells in the hepatic tissue sections of nude mice.But the control group did not show PKH26 labeled cells.The expressions of ALB were positive in the experimental group as detected by immunocytochemical staining after two weeks of the cells transplantation.ConclusionHuman peripheral blood mononuclear cells have the potential to differentiate into hepatocyte-like cells under the induction of HGF and FGF-4.Additionally,PKH26 is an effective tracer in hepatocyte-like cell transplantation.The hepatocyte-like cells settled in hepatic tissue begin to differentiate into mature hepatocyte after two weeks of the cells transplantation.It plays hepatic cells function and expresses alhnmin.

Objective To investigate the specific sites that estrogen receptor (ER)α could be recruited to in the p21WAF1/CIP1 promoter region to regulate its transcriptional activity in MCF-7 cells,and to clarify the molecular mechanism of suberoylanilide hydroxamic acid (SAHA) and leptin in the regulation of p21WAFI/CIP1 promoter function.Methods MCF-7 cells were starved by culturing them in fetal calf serum-free medium for 24 hours,and then treated with 20 μmol/L of 0.88 μL SAHA (SAHA group) or 0.625 nmol/L of 10 μL leptin (leptin group) for 24 hours,or cultured in complete RPMI-1640 medium (control group).Cell lysates were incubated with anti-ERα antibody for ChIP analysis.The relative expression levels of DNA fragments,ranging from the TSS to upstream of the p21WAF1/CIP1 promoter region (+2 to-4 000 bp),that bound the antibody were detected by real-time PCR.Results In the control group,the relative expression levels of f1,f2,and f8 DNA fragments that bound the antiERα antibody were two-fold higher than the relative expression of the f9 fragment (P ＜ 0.01).In the SAHA and leptin groups,the relative expression of f1 to f10 DNA fragment that bound anti-ERα antibody was significantly lower than that of the control.The binding affinity of ERα for the f8 fragment was the lowest (P ＜ 0.01) in the SAHA group,and it was significantly lower than that in the leptin group (P ＜ 0.01).Conclusion ERα could be recruited to the p21WAFI/CIP1 promoter via signaling pathways activated during the proliferation of breast cancer MCF-7 cells.Moreover,the DNA fragment ranging from-2 800 to-3 200 bp upstream of the p21 WAF1/CIP1 promoter is the target functional region for high-affinity binding with ERα.

Traditional Chinese medicine,ancient Greek medicine,Ayurvedic medicine,and Arab medicine are recognized as the four major traditional medicines in the world.It reviews the education and training systems of the four major traditional medicines and finds that traditional Chinese medicine focuses on the teacher-student relation-ship and the combination of theory and practice;Ancient Greek medicine was mainly characterized by strong theoreti-cal research and experimental observation;Ayurveda highly values cultural identity as its main characteristic;Arab medicine attaches great importance to cultural exchange and practical promotion.It suggests promoting innovative de-velopment,strengthening practical teaching,improving teaching quality,strengthening international exchanges and cooperation,and increasing public acceptance abroad.

Traditional Chinese medicine,ancient Greek medicine,Ayurvedic medicine,and Arab medicine are recognized as the four major traditional medicines in the world.It reviews the education and training systems of the four major traditional medicines and finds that traditional Chinese medicine focuses on the teacher-student relation-ship and the combination of theory and practice;Ancient Greek medicine was mainly characterized by strong theoreti-cal research and experimental observation;Ayurveda highly values cultural identity as its main characteristic;Arab medicine attaches great importance to cultural exchange and practical promotion.It suggests promoting innovative de-velopment,strengthening practical teaching,improving teaching quality,strengthening international exchanges and cooperation,and increasing public acceptance abroad.

Traditional Chinese medicine,ancient Greek medicine,Ayurvedic medicine,and Arab medicine are recognized as the four major traditional medicines in the world.It reviews the education and training systems of the four major traditional medicines and finds that traditional Chinese medicine focuses on the teacher-student relation-ship and the combination of theory and practice;Ancient Greek medicine was mainly characterized by strong theoreti-cal research and experimental observation;Ayurveda highly values cultural identity as its main characteristic;Arab medicine attaches great importance to cultural exchange and practical promotion.It suggests promoting innovative de-velopment,strengthening practical teaching,improving teaching quality,strengthening international exchanges and cooperation,and increasing public acceptance abroad.

Traditional Chinese medicine,ancient Greek medicine,Ayurvedic medicine,and Arab medicine are recognized as the four major traditional medicines in the world.It reviews the education and training systems of the four major traditional medicines and finds that traditional Chinese medicine focuses on the teacher-student relation-ship and the combination of theory and practice;Ancient Greek medicine was mainly characterized by strong theoreti-cal research and experimental observation;Ayurveda highly values cultural identity as its main characteristic;Arab medicine attaches great importance to cultural exchange and practical promotion.It suggests promoting innovative de-velopment,strengthening practical teaching,improving teaching quality,strengthening international exchanges and cooperation,and increasing public acceptance abroad.