VEGF and Notch signaling pathways are important mechanisms in regulation of embryonic vascular development and tumor angiogenesis.Blockade of the VEGF pathway effectively inhibits tumor anglo- genesis and growth.Recent findings indicate that Notch signaling decreases angiogenesis by suppressing endo-thelial tip cell formation.Combination therapy by blocking Notch and VEGF pathways synergistically inhibits tumor growth in preclinical models.Thus,targeting VEGF and Notch pathways may lead to new therapies for clinical application.

Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts (COE) on vascular endothelial growth factor (VEGF) mRNA and protein expression in hepatoma (Hepal-6) cells of mice.Methods Hepa1-6 cells were treated with COE at different nontoxic concentration (0, 10, 20, 40, 80, and 160 μg/mL) for 16 h. The mRNA and protein expressions of VEGF were detected by reverse transcription-PCR and Western Blotting,respectively. Results COE significantly inhibited VEGF expression at both mRNA and protein levels in a dose-dependent manner. Conclusion COE can inhibit VEGF expression in Hepa1-6 cells, therefore suggest that VEGF could be chosen as an therapeutic target for COE in the context of cancer chemoprevention and anticancer therapy.

As a proto-oncogene, urothelial carcinoma antigen 1 (UCA1) is highly expressed in many human tumors such as bladder cancer, gastric cancer, breast cancer, ovarian cancer and brain glioma, which shows important application value in the diagnosis, treatment and prognosis of tumor.Studies show that UCA1 can promote tumor cell proliferation through multiple signaling pathways and molecular mechanisms, which may become a new potential target for the diagnosis and treatment of tumor.

Objective:To analyze the frequencies of HLA-A*0201 restricted CEA-specific CD8+T cells, HLA-A*0201/FLUmp tetramer and HLA-A*0201/CAP-1 tetramer were applied in patients with colorectal cancer.Methods: Lymphocytes from peripheral blood and lymph node,1×106 cells/ml,were incubated with 1μg HLA-A*0201/peptide tetramers and anti-CD8 for 1 h at 25 coseperately.The cells were then washed in PBS.Next,the cells were illuminated by detecting frequencies of FLUmp-specific CD8+T cells and CAP-1-specific CD8+T cells with flow cytometry.Results: HLA-A*0201/peptide were used to detect CAP-1 or FLUmp-specific CD8+T cells,which were analyzed either healthy individuals or patients with colorectal cancer.We did not find differences in average frequencies of FLUmp-specific CD8+T cells between 11 HLA-A*0201+patients with colorectal cancer and 14 HLA-A*0201+healthy individuals [ ( 0.671 ±0.421 )%, ( 0.564 ±0.408 )%].But the frequencies of CAP-1-specific CD8+T cells of HLA-A*0201+patients with colorectal cancer showed higher than HLA-A*0201+healthy individuals [ ( 2.409 ± 2.385 )%, ( 0.020 ± 0.021)%respectively],which was statistically significant(P=0.008).Conclusion:The frequencies of CAP-1-specific CD8+T cells in PBMC from peripheral blood and lymph node of HLA-A*0201+patients were increased,showed CEA-specific CTs has a vital role in colorectal cancer.

Objective To examine the immunoregulation of Celastrus orbiculatus extracts(COE),a traditional Chinese medicine,on maturation and function of dendritic cells(DCs)in vitro and in vivo.Methods In vitro,after treated with COE indifferent nontoxic concentrations(0,10,20,40,80,and 160 μg/mL)for 5 d,the surface immunological molecules andcytokine secretion of mice bone marrow-derived DCs in response to COE were analyzed by flow cytometric analysis(FACS)and enzyme linked immunosorbent assay(ELISA),respectively.In vivo,mouse hepatoma cells(Hepal-6,1 ×106)were injected sc and were treated with different dosages of COE(10,20 or 40 mg/kg/d).Effects on tumor growth were determined by tumor volume and histology analysis after 28 d administration of COE.The relative proportions ofmature DCs and CD8+ T cells were measured in mononuclear cells that had been isolated from spleen by FACS.Results COE stimulated IL-2 and IFN-γ secretion of DCs,simultaneously enhanced the maturation of DCs byenhancing immunological molecule(CD40,CDS0,CD86,H-2Kb,and I-Ab)expression in a dose-dependent manner.Furthermore,the chcmotactic responses of DCs were significantly higher in COE-treated than untreated DCs,in association with higher chcmokine receptor 7 expression.Furthermore,COE increased DCs produce IFN-γ and IL-2 ina dose-dependent manner when the concentration of COE less than 40 μg/mL,decreased DCs produce IL-10 and IL-4also in a dose-dependent manner.In in vivo studies,COE can not only suppress growth of malignant hepatocellularcarcinomas but also stimulate maturation of DCs,associated with strongly enhanced CD8+ CTL responses.ConclusionThese data provide new insight into the mechanism of action of COE and indicate that the stimulation of maturation andfunction of DCs by COE contributes to its immunoregulatory effects.

Heat shock protein 27 ( HSP27 ) is an endogenous protein that plays an important role in a great variety of physio-logical and pathological processes. It can express a large number under body stress conditions. Recent studies have shown estro-gen upregulates the expression of HSP27 through a number of ways, playing a perfect “triple protection” role. In the early stage of atherosclerosis, estrogen induces the phosphorylation of HSP27 via PI3K/Akt signaling pathway. Phosphorylation of HSP27 can resist the injury of vascular endothelial cells( VECs) through an antioxidant and anti-apoptotic pathway as well as the inhibition of cytochrome C. In the stage of forming foam cells, estrogen induces the expression and release of HSP27 from mac-rophages by stimulating the estrogen receptor β ( ERβ) , then HSP27 inhibits the LDL uptake and the release of proinflammato-ry cytokine by binding scavenger receptor A ( SR-A) . During the proliferation and migration of vascular smooth muscle cells ( VSMCs) , estrogen induces estrogen receptor α ( ERα) and protein phosphatase 2 ( PP2A) to form a complex that enhances the activity of PP2A, then it can lead to the dephosphorylation of HSP27 and finally inhibit cells proliferation and migration. In summary, the anti-atherosclerotic effect of estrogen is closely re-lated to the role of HSP27. Given the side effects of estrogen re-placement therapy( MHT) , regulating HSP27 may provide a no-vel therapy for the prevention and treatment of cardiovascular dis-eases in menopausal women clinically.

This study aimed at investigating the effects of the extract of Chinese herb,Nansheteng (C.orbiculatus Thunb.),on human HepG2 cells through the overexpression of mTOR.The GV238-mTOR recombinant plasmids were transfected into HepG2 cells using molecular biological technology.The expression level of mTOR was evaluated by means of relative activity of luciferase and western blot.Human hepatic carcinoma HepG2/mTOR++ cells were treated with C.orbiculatus extract in different concentrations (20,40,80,160 and 320 tg·mL-1) for 24 h.The mTOR protein expression was detected by western blot.It was found that the protein expression of mTOR in transfected HepG2 cells was significantly enhanced.C.orbiculatus extract significantly inhibited the proliferation of HepG2/mTOR++ cells.Simultaneously,C.orbiculatus extract inhibited mTOR at its protein level in a dose-dependent manner.In conclusion,we successfully constructed recombinant mTOR cloning vectors,and established the stable HepG2 cell line with the overexpression of mTOR.Besides,C.orbiculatus extract significantly inhibited mTOR protein expression in human hepatic carcinoma HepG2 cells.

Cross-talk of intracellular signaling pathways that share common components (hubs) is organized in form of a bow-tie network topology.Signaling cross-talk is functionally pleiotropic for target genes regulation, resulting in functional redundancy, synergism and antagonism, which should be precisely controlled to prevent signaling 'leaking' or 'spillover'.Thus, the biological system has evolved multiple insulating mechanisms to achieve stimulus-specific response that maintains intracellular homeostasis.The insulation mechanism of signaling cross-talk suggests: (1) the functional duality of cross-talk molecules that determine cell fate requires selectively targeting dysregulated cross-talk molecules while protecting the normal ones from off-target or unintended effects, and we propose them as the targetable cross-talk molecules;(2) cross-talk molecules are usually carried on the macromolecular complex as their functional platforms, thus the structural plasticity of conformational changes at the interaction surface of cross-talk molecules asks for intensive work on the relationship study between drug binding and biological activity, which we propose as the accessible cross-talk molecules.Therefore, signaling cross-talk and its insulation mechanism play instructive leading roles in resolving the bottlenecks of current drug R&D and improve the clinical outcome.

胰岛素样生长因子（IGF）系统是人体内的一组多肽类物质，其分泌细胞广泛分布在人体各组织中，具有促生长作用。 IGF系统能够诱导肿瘤细胞出现EMT，进而增强肿瘤细胞干性、自我更新和转移潜能，向恶性肿瘤发展。研究显示，IGF系统的过 表达与胃癌（GC）的发生存在着密切关联，例如IGF-1和IGF-1R在GC组织中就有着明显表达，其表达水平随着GC从良性到恶性 的转变而逐渐増强。近年来研究也表明， 在GC细胞癌变的过程中，EMT起着关键性的作用，它使得肿瘤细胞具备了侵袭和迁移 能力，而削弱和抑制这种能力则成为了一种有针对性的治疗方法，这其中有着很大的研究空间。临床数据显示，患者化疗前后血 清中的IGF-1表达水平也能够直接反映化疗的效果，这更显示IGF可作为GC筛查的重要标志物， 为GC精准治疗的研究提供可 靠依据。上述一系列发现，使得IGF有望成为新的判断肿瘤发生及转移的诊疗指标。

This study was aimed to investigate the effect of Nan-She-Teng (Celastrus orbiculatus) extract on epithe-lial-mesenchymal transition(EMT) in HepG2 cells. Except the control group, human hepatocellular carcinoma HepG2 cells in other groups were treated with Celastrus Orbiculatus extract in different concentrations (10, 20, 40, 80, and 160 μg·mL-1). The protein expression levels related to EMT were detected by western blotting. At 48 h after fertiliza-tion, the zebrafish embryos were randomly assigned to 7 groups as follows: untreated control group (E3 buffer), DMSO group (E3 buffer with 1% DMSO), and different dosages treatment of C.orbiculatus extract (10, 20, 40, 80, and 160μg·mL-1) for 24 h. The protein expressions of mTOR signaling pathways were detected by western blotting. The re-sults showed that compared with the control group, C.orbiculatus extract significantly increased E-cadherin protein expression. Simultaneously, C.orbiculatus extract inhibited vimentin and mTOR signaling pathways at protein levels. It was concluded that to a certain extent, C.orbiculatus extract prevented EMT in HepG2 cells by modulating the mTOR signaling pathway. Therefore, it suggested that mTOR can be chosen as a new therapeutic target for clinical treatment of hepatic carcinoma.