Objective:To investigate the effect of verbascoside(VERB)on high-fat diet-induced atherosclerosis(AS)in ApoE-/-mice and the effect on high mobility histone 1(HMGB1)/receptor for glycation end products(RAGE)/nuclear factor κB(NF-κB)pathway.Methods:A total of 90 ApoE-/-mice were randomly divided into normal group,AS group,VERB group,simvastatin group and VERB+pathway activator HMGB1 group,with 18 mice per group.After 8 weeks of group administration,blood and aorta samples were taken.Fasting serum triacylglycerol(TG),total cholesterol(TC)and low density lipoprotein(LDL)levels were deter-mined by automatic biochemical analyzer.Oil red O,HE and TUNEL staining were performed to observe apoptosis of aortic plaque and endothelial cell(EC).Flow cytometry was performed to analyze circulating EC numbers.Immunohistochemistry was performed to analyze the infiltration area of macrophages(CD68+)and T lymphocytes(CD3+)in aortic plaques.Western blot was performed to detect expressions of HMGB1/RAGE/NF-κB pathway,inflammation and adhesion molecules.Results:Compared with normal group,AS group had lipid plaques in arterial intima,thickness of the media was uneven,TG,TC,LDL levels,lession proportion,plaque area,circulating EC number,arterial EC apoptosis rate,macrophage(CD68+)and T lymphocyte(CD3+)infiltration areas,TNF-α,monocyte chemoattractant protein-1(MCP-1),vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),cytoplasm HMGB1,total HMGB1,total RAGE protein levels and nuclear/total p65 NF-κB levels were increased(P<0.05),while nuclear HMGB1 protein,nuclear/total HMGB1 and cytosolic p65 NF-κB levels were decreased(P<0.05).After VERB or simvastatin intervention,arterial lesions were alleviated,TG,TC,LDL levels,lession proportion,plaque area,circulating EC number,arterial EC apoptosis rate,macrophage(CD68+)and T lymphocyte(CD3+)infiltration areas,TNF-α,MCP-1,VCAM-1,ICAM-1,cytoplasm HMGB1,total HMGB1,RAGE protein levels and nuclear/total p65 NF-κB level were decreased(P<0.05),while nuclear HMGB1 protein,nuclear/total HMGB1 and cytosolic p65 NF-κB levels were increased(P<0.05),and HMGB1 was able to antagonize the protective effect of VERB on AS mice.Conclusion:VERB can inhibit expressions of inflammatory and adhesion fac-tors in arterial plaques in ApoE-/-mice,reduce EC shedding and apoptosis,therefore improve AS symptoms in ApoE-/-mice,and the mechanism may be related to the inhibition of HMGB1/RAGE and NF-κB pathway.

Objective:To investigate the effect of verbascoside(VERB)on high-fat diet-induced atherosclerosis(AS)in ApoE-/-mice and the effect on high mobility histone 1(HMGB1)/receptor for glycation end products(RAGE)/nuclear factor κB(NF-κB)pathway.Methods:A total of 90 ApoE-/-mice were randomly divided into normal group,AS group,VERB group,simvastatin group and VERB+pathway activator HMGB1 group,with 18 mice per group.After 8 weeks of group administration,blood and aorta samples were taken.Fasting serum triacylglycerol(TG),total cholesterol(TC)and low density lipoprotein(LDL)levels were deter-mined by automatic biochemical analyzer.Oil red O,HE and TUNEL staining were performed to observe apoptosis of aortic plaque and endothelial cell(EC).Flow cytometry was performed to analyze circulating EC numbers.Immunohistochemistry was performed to analyze the infiltration area of macrophages(CD68+)and T lymphocytes(CD3+)in aortic plaques.Western blot was performed to detect expressions of HMGB1/RAGE/NF-κB pathway,inflammation and adhesion molecules.Results:Compared with normal group,AS group had lipid plaques in arterial intima,thickness of the media was uneven,TG,TC,LDL levels,lession proportion,plaque area,circulating EC number,arterial EC apoptosis rate,macrophage(CD68+)and T lymphocyte(CD3+)infiltration areas,TNF-α,monocyte chemoattractant protein-1(MCP-1),vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),cytoplasm HMGB1,total HMGB1,total RAGE protein levels and nuclear/total p65 NF-κB levels were increased(P<0.05),while nuclear HMGB1 protein,nuclear/total HMGB1 and cytosolic p65 NF-κB levels were decreased(P<0.05).After VERB or simvastatin intervention,arterial lesions were alleviated,TG,TC,LDL levels,lession proportion,plaque area,circulating EC number,arterial EC apoptosis rate,macrophage(CD68+)and T lymphocyte(CD3+)infiltration areas,TNF-α,MCP-1,VCAM-1,ICAM-1,cytoplasm HMGB1,total HMGB1,RAGE protein levels and nuclear/total p65 NF-κB level were decreased(P<0.05),while nuclear HMGB1 protein,nuclear/total HMGB1 and cytosolic p65 NF-κB levels were increased(P<0.05),and HMGB1 was able to antagonize the protective effect of VERB on AS mice.Conclusion:VERB can inhibit expressions of inflammatory and adhesion fac-tors in arterial plaques in ApoE-/-mice,reduce EC shedding and apoptosis,therefore improve AS symptoms in ApoE-/-mice,and the mechanism may be related to the inhibition of HMGB1/RAGE and NF-κB pathway.

Objective To investigate the expression of SLC35A2 and PFDN2 in breast cancer and their relationship with clinical indicators and prognosis.Methods TCGA database and TIMER 2.0 database were used to analyze the differences of SLC35A2 and PFDN2 expression in breast cancer tissues and paracancerous tissues;K-M database was used to create the survival curves of patients in the high and low expression groups of the two.qRT-PCR and immunohistochemistry were used to detect the expression of SLC35A2 and PFDN2 in the cancerous and paracancerous tissues,and the expression differences,the relationship between their expression levels and the clinical observation indexes were statistically analyzed,and the independent prognostic factors of breast cancer were screened out;K-M survival analysis was used to compare the prognostic differences between the groups and create the survival curves.Results The expression levels of SLC35A2 and PFDN2 in breast cancer tissues were significantly higher than those in paracancerous tissues according to the results of biopsy,qRT-PCR and immuno-histochemistry,and the expression levels of SLC35A2 were significantly correlated with lymph node metastasis,while the expression of PFDN2 was significantly correlated with the diameter of the tumor and the metastasis of lymph nodes,and the expression of SLC35A2 and PFDN2 was an independent prognostic factor for breast cancer.patients had the worst prognosis.Conclusion The expression of SLC35A2 and PFDN2 in breast cancer tissues was closely related to clinical indicators and prognosis of breast cancer,and could be used as a potential target for breast cancer treatment.

Objective To evaluate the effects of artemisinin on proliferation,cell cycle and apoptosis of gallbladder cancer cells.Methods Gallbladder carcinoma cell lines (GBC-SD and NOZ) were cultured in vitro.The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay.The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 μ mol/L) were examined using flow cytometry.The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 μ mol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2,CDK4,cyclin D1,p16,cytochrome C and caspase-3 were examined by Western blot assay.t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups,respectively.Results The cell proliferation was significantly inhibited by artemisinin,the ICS0 of artemisinin against GBC-SD and NOZ cells were 14.05 μmol/L and 12.42 μmol/L,respectively.Artemisinin induced cycle arrest,and G1 population of GBC-SD and NOZ cells increased to 74.60％ and 78.86％.Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67％ and 16.51％ after dealt with artemisinin,respectively.In addition,expression of pl6 was increased,and expressions of p-ERK1/2,CDK4 and cyclin D1 were down-regulated by artemisinin(all P ＜0.05).Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin (P ＜ 0.05).Conclusion The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway,G1 phase arrest and triggering caspase-3-mediate apoptosis.

Objective To evaluate the effects of artemisinin on proliferation,cell cycle and apoptosis of gallbladder cancer cells.Methods Gallbladder carcinoma cell lines (GBC-SD and NOZ) were cultured in vitro.The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay.The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 μ mol/L) were examined using flow cytometry.The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 μ mol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2,CDK4,cyclin D1,p16,cytochrome C and caspase-3 were examined by Western blot assay.t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups,respectively.Results The cell proliferation was significantly inhibited by artemisinin,the ICS0 of artemisinin against GBC-SD and NOZ cells were 14.05 μmol/L and 12.42 μmol/L,respectively.Artemisinin induced cycle arrest,and G1 population of GBC-SD and NOZ cells increased to 74.60％ and 78.86％.Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67％ and 16.51％ after dealt with artemisinin,respectively.In addition,expression of pl6 was increased,and expressions of p-ERK1/2,CDK4 and cyclin D1 were down-regulated by artemisinin(all P ＜0.05).Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin (P ＜ 0.05).Conclusion The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway,G1 phase arrest and triggering caspase-3-mediate apoptosis.

Based on the theory of cancer stem cells (CSCs),people have been searching for the treatments of malignant cancers.Gastric cancer side population cells (SP) have the characteristics of CSCs.Searching for the molecular targeted therapy strategy of gastric cancer which embarks from the gastric cancer SP and is based on the theory of CSCs provides a new direction for the treatment,early diagnosis,therapeutic effect and prognosis of gastric cancer.