ObjectiveTo explore the imaging of the thrombosis after pharmacological triggering of plaque rupture in atherosclerotic rabbit model by using 3.0 T high-resolution magnetic resonance imaging.MethodsTwenty male New Zealand white rabbits were divided into an experimental group (n = 16) and a control group (n = 4).The aortic wall injuries were induced by an intravascular balloon in experimental group rabbits after high cholesterol diet.The pharmacological triggering with Russell's viper venom and histamine was performed after 3 months of establishment of model.All of the animals underwent pre-trigger and post-trigger MR examinations including 3D time of fight (3D TOF),T1 WI,T2WI and post contrast T1 WI.Euthanasia was performed in all rabbits and gross anatomy and histological specimen of aorta were obtained.Comparing the location and length of the thrombus between MRI images and histopathology was used Pearson test.Comparing the calculated indexes of abdominal aorta between rabbits with and without thrombosis was used AVONA test and LSD-t test.Results After triggering,8 in 14 survived rabbits developed thrombosis in experimental group,meanwhile,no thrombus was found in control group.The accuracy of multi-sequences MRI for detecting of thrombus was 87.1％ (27/31).MRI data correlated with the histopathology regarding thrombus length ( r = 0.85,P ＜ 0.01 ) and thrombus location ( r = 0.94,P＜0.01 ).Compared with rabbits without thrombosis,the rabbits with thrombosis had narrower lumen of abdominal aorta in the pre-triggered MR images [ ( 5.71 ± 2.38 )mm2 vs.( 8.93 ± 5.36) mm2,P ＜ 0.01 ].ConclusionMRI is useful tool to determine the thrombosis and plaque rupture in atherosclerotic rabbit model.

OBJECTIVETo investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells.

METHODSThe multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR.

RESULTSThe drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells.

CONCLUSIONERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Phosphorylation ; Up-Regulation

Medical students differ greatly in their thinking and cognition of the value and pursuit of life existence. The mainstream values are positive and pragmatic, but they are also characterized by decentralization, diversification, individualization and variability. Value education needs to be object-centered, take care of students’ individual lives, understand and answer their real questions to be more accurate and effective. In the intelligent technology society, the education of life value of medical students in the ideological and political courses should rely on the intelligent technology platform to achieve the precise coupling of object and value goal through the form of question exchange. It is necessary to transcend the limitations of medical science perspective, realize accurate life value education, face the nihilistic challenge of life value, understand the systematic and dialectical significance of life events, and return to the original intention and mission of medical science to take care of life itself.

Objective To investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells. Methods The multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR. Results The drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells. Conclusion ERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

Objective To investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells. Methods The multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR. Results The drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells. Conclusion ERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

Objective To analyze the combination characteristics between Tir-IBD( intimin binding domain) and its ligand intimin or mutant intiminN916Y of EHEC O157 ∶H7.Methods The gene of TirIBD (tir-ibd) from EHEC O157 ∶H7 chromosome was cloned into pMD18-T vector.Thereafter,the amplified gene was cloned into prokaryotic expression plasmid pET-21 a (+).The recombinant pasmid pET-21 a( +)-tir-ibd was transformed into E.coli BL21 (DE3).After inducement,the protein Tir-IBD was successfully expressed and analyzed with SDS-PAGE and Western blot.It was purified by affinity chromatography and ionexchange chromatography and was coupled on the Ni-NTA chip of BIACore 3000.Then the ligand intimin and mutant intiminN916Y were flow through the chip and their combination characteristics were detected.Results In the present study,the gene of Tir-IBD(tir-ibd) was successfully cloned into pET-21a(+).The results of SDS-PAGE and Western blot assay showed that the protein was successfully expressed,which accounts for 15％ of total expression products,and its molecular weight was about 10×103.The purity was above 95％ after purification.After coupled on the Ni-NTA chip of BIACore 3000,their combination characteristics with ligand intimin and mutant intiminN916Y were successfully detected.The equilibrium binding constants Ka was obtained by fitting the data with the BIACore evaluation program ( Version 4.1 ).The result showed that the combination characteristics between Tir-IBD and intimin have some difference compared with that of mutant intiminN916Y and the difference is temperature dependent.Conclusion Tir-IBD of EHEC O157 ∶H7 was successfully constructed and purified.The method to analyze the combination characteristics between Tir-IBD and its ligand intimin or mutant was established.The combination characteristics between Tir-IBD and intimin or mutant intiminN916Y have some temperature dependent difference and the mutated amino acids residue is crucial for their receptor-ligand binding.

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.