Objective To explore the anti-inflammatory effect of erythropoietin (EPO) on hepatic ischemia reperfusion (IR) injury in fatty liver rats.Methods A total of 100 male SD rats were fed with high-fat diet for 12 weeks.After the model was successfully established,the fatty liver rats were randomly divided into sham-operation (SHAM),the ischemia-reperfusion (IR) and EPO preconditioning group.Serum ALT and AST as well as hepatic histopathological changes were measured.Xanthine oxidase method was used to detect the liver tissue SOD.Thiobarbituric acid method was used to detect the MDA.Enzyme-linked immunosorbent adsorption assay (ELISA) was used to detect the plasma tumor necrosis factor alpha (TNFαt) and interleukin 1 (IL-1).Results In the EPO preconditioning groups the swelling hepatocytes was observed,but the inflammatory cell infiltration was significantly decreased and no necrosis of hepatocytes was found.ALT and AST in the EPO preconditioning groups were significantly lower than those in IR group (P ＜ 0.05).The levels of SOD activity in the EPO preconditioning groups were significantly higher,but MDA were significantly lower than that in IR group (P ＜ 0.05).The TNF-α and IL-1 in the EPO preconditioning groups were significantly lower than those in IR group (P ＜0.05).The values of ALT,AST,TNF-α,IL-1 and MDA in the EPO groups were:EPO-1 ＞ EPO-2 ＞ EPO-3 (P ＜ 0.05);but the values of SOD in the EPO groups were:EPO-1 ＜ EPO-2 ＜ EPO-3 (P ＜ 0.05).Conclusions EPO preconditioning has a protective effect against hepatic IR injury in fatty liver rats,possibly through inhibiting the inflammatory reaction to prevent the IR injury.The anti-inflammatory effect of high-dose EPO is better than that of low-dose EPO.

Adenoma, Liver Cell ; Gallbladder Diseases ; Histiocytosis, Sinus ; Humans ; Liver Neoplasms

Objective To investigate the change of the percentage of Th17 cells and cytokine among hepatocellular carcinoma (HCC) tissues and para-carcinoma tissue and the liver tissue of hepatic hemangio-ma, and to explore its clinical significance. Methods To choose 26 of hepatocellular carcinoma patients between November 2014 and December 2016 and also include 11 cases of hepatic hemangioma as control group. The percentage of Th17cells in HCC tissue and para-carcinoma tissue and the liver tissue of hepatic hemangioma was evaluated by flow cytometric; The levels of IL-6 and IL-17 in HCC tissues, para-carcinoma tissue and the liver tissue of hepatic hemangioma were detected by RT-PCR. Results The ratio of Th17 cells to CD4 cells in peripheral blood of HCC was significantly higher than that in control group (4. 07% ± 0. 68% vs. 1. 39% ± 0. 41% ), P＜0. 05. The ratio of Th17 cells to CD4 cells in patients with hepatocellular carcinoma was significantly higher than that in paracancerous tissues and liver tissues of hepatic hemangioma (4. 76% ± 0. 75% vs. 2. 30% ± 0. 26% vs. 0. 77% ± 0. 31% , P＜0. 05). Multivariate linear stepwise regression analysis of the changes of Th17 cells in patients with HCC and their correlation with clinical parameters showed that Th17 lymphocytes in peripheral blood of patients with HCC were closely related to TNM staging(YTh17 =2. 647 +0. 687TNM. It’s indicated that the level of IL-6 and IL-17 in liver cancer tissues was significantly higher than that of the liver hemangioma in the control group through ELISA ( P＜0. 05). The proportion of peripheral blood Th17 cells to CD4 cells is different in different TNM stages(stage I＜stage II ＜stage III ～IV, the difference was statistically significant. RT-PCR analysis showed that the expression of IL-17, IL-6mRNA in hepatocellular carcinoma was significantly higher than that in adjacent tissues and hepatic hemangioma liver tissue (all P＜0. 05). Conclusion The increased proportion of Th17 cells and the increased levels of cytokines IL-6 and IL-17 may be closely correlated with occurrence and development of hepatocellular carcinoma. It can be an important target of anti-tumor therapy.

Objective:To evaluate the effect of down-regulation of Sp1 expression on the radiosensitivity of glioma cells.Methods:The oligonucleotide sequence encoding shRNA was designed and synthesized, and cloned into LV3 (H1/GFP & Puro) vector to construct the recombinant. U251 and U87 cells were infected with recombinant lentivirus, then the stably-transfected cell lines were obtained by puromycin screening. The expression levels of Sp1 mRNA and protein were detected by RT-PCR and Western blot. Cell survival was detected by clonal survival assay, cell cycle was determined by flow cytometry, and DNA damage was measured by immunofluorescence assay, respectively.Results:At 72 h after infection, high expression of Sp1 lentiviral vector was observed in two cell lines under fluorescence microscope. RT-PCR and Western blot confirmed that the expression levels of Sp1 mRNA and protein were significantly down-regulated in both transfected cells (both P<0.01) and the silencing rates of Sp1 were above 90%. The sensitization enhancement ratio (SER) of shRNA-U251 and shRNA-U87 cells at 10% cell survival level were 1.39 and 1.18, respectively. After irradiation, the G 2/M phase ratio and the number of γ-H2AX foci in two Sp1 knockout groups were significantly increased. Conclusion:shRNA silencing of Sp1 increases the G 2/M phase arrest induced by X-ray, aggravates the degree of DNA double-strand breaks, and improves the radiosensitivity of glioma cells.

KRAS‒PDEδ interaction is revealed as a promising target for suppressing the function of mutant KRAS. The bottleneck in clinical development of PDEδ inhibitors is the poor antitumor activity of known chemotypes. Here, we identified novel spiro-cyclic PDEδ inhibitors with potent antitumor activity both in vitro and in vivo. In particular, compound 36l (K _D = 127 ± 16 nmol/L) effectively bound to PDEδ and interfered with KRAS-PDEδ interaction. It influenced the distribution of KRAS in Mia PaCa-2 cells, downregulated the phosphorylation of t-ERK and t-AKT and promoted apoptosis of the cells. The novel inhibitor 36l exhibited significant in vivo antitumor potency in pancreatic cancer patient-derived xenograft (PDX) models. It represents a promising lead compound for investigating the druggability of KRAS‒PDEδ interaction.