Objective: To investigate the effect of activation of the stimulator of interferon genes(STING) pathway on macrophage polarization function and its role in T-cell response. Methods: Mouse macrophage RAW264.7 cells were used.STING signaling related proteins in RAW264.7 macrophage treated with STING agonist diABZI were analyzed by Western blot,including TANK-binding kinase-1(TBK1),interferon regulatory factor-3(IRF3),STING,p-TBK1,p-IRF3,p-STING.The polarization of macrophage RAW264.7 cells treated with diABZI was analyzed by flow cytometry.Co-culture of diABZI-treated RAW264.7 macrophage and T cells was applied to evaluate the change of T cell response. Results: STING signaling related proteins were upregulated in macrophage RAW264.7 cells treated with diABZI for 3 hours.The expression of CD86 was upregulated on the surface of macrophages after 12 hours of diABZI treatment,and the CD86/CD206 ratio was elevated,which presented the M1 polarization phenotype.When coculturing diABZI-treated macrophage RAW264.7 cells with T cells,the cytokine secretion ability of T cells including CD4+T and CD8+T cells was enhanced and the expression of CD107a in CD8+T cells was upregulated. Conclusion STING signaling induces M1 polarization of macrophages which enhance the function of T cells,especially CD8+T cell immune response.

Objective To investigate the role and mechanism of the DNA helicase PIF1 in the proliferation of ovarian cancer cells and its response to DNA damage.Methods The relative expression of the PIF1 gene in normal ovarian tissue compared to ovarian cancer tissue,as well as the relationship between PIF1 expression and overall survival in ovarian cancer patients,was analyzed using the Gene Expression Profiling Interactive Analysis(GEPIA)and Kaplan-Meier Plotter public databases.PIF1 knockout ovarian cancer cells were established using CRISPR/Cas9 gene-editing technology.A retroviral vector overexpressing Rad51 recombinase was constructed,and then transfected into PIF1 knockout ES-2 ovarian cancer cells.Western blot analysis was used to determine the effects of PIF1 knockout and Rad51 overexpression in the transfected cells.Cell proliferation was assessed with cell counting,colony formation assay and CCK-8 assay.A total of 32 female BALB/c nude mice(6～8 weeks old,weighing 20～25 g)were randomly divided into ES-2 control group,ES-2 knockout group,OVCAR-3 control group,and OVCAR-3 knockout group,with 8 mice in each group.A mouse xenograft model was established to assess the in vivo proliferative capacity of ovarian cancer cells.Apoptotic rate and cell cycle were assessed using flow cytometry.The senescence of ovarian cancer cells was evaluated through a β-galactosidase activity assay.Western blotting and immunofluorescence assay were applied to determine the changes in phosphorylated histone H2AX(γ-H2AX)protein were measured with to evaluate the effects of PIF1 knockout and Rad51 overexpression on DNA damage and to observe the localization of PIF1 and Rad51 in ovarian cancer cells.Results The analysis of public databases revealed that PIF1 overexpression was negatively correlated with the overall survival of patients(P＜0.001),and PIF1 was found to be overexpressed in the ovarian cancer tissues than the normal ovarian tissues(P＜0.05).CRISPR/Cas9-mediated knockout of PIF1 significantly inhibited the proliferation(P＜0.01)and clonogenic ability(P＜0.001)of ovarian cancer cells,which was also validated in mouse model(P＜0.05).Flow cytometry indicated that PIF1 knockout promoted apoptosis(P＜0.01)and induced cell cycle arrest(P＜0.01)in ovarian cancer cells.In addition,β-galactosidase activity assay demonstrated that PIF1 knockout enhanced cellular senescence(P＜0.001).Western blot and CCK-8 assays further revealed that PIF1 knockout increased the expression of γ-H2AX protein(P＜0.05)and suppressed the proliferative capacity of ovarian cancer cells following cisplatin treatment(P＜0.05).In PIF1 knockout ovarian cancer cells,Rad51 expression was diminished.However,overexpression of Rad51 in PIF1-deficient cells restored PIF1 expression,decreased γ-H2AX protein level(P＜0.05),and rescued cell proliferation(P＜0.01).Immunofluorescence assay demonstrated that EGFP-PIF1 and Rad51 were co-localized in the nucleus.Conclusion PIF1 and Rad51 collaboratively regulate DNA damage and cell proliferation in ovarian cancer cells.

Objective: To construct the mmu-miR-155 eukaryotic overexpression vector pmR-155 and to investigate its effect on HBV replication and expression of PTEN in vivo. Methods: The mmu-mir-146a precursor gene fragment pre-mmu-mir-146a was amplified by PCR, then connected to the pmR-mCherry plasmid vector after double enzyme digestion, the accuracy of recombinant vector was verified by colony PCR、double enzyme digestion and sequencing; then the recombinant vector was transfected HBV transgene mice(Experimental Group)with hydrodynamics-based injection via vena caudalis, and pmR-mCherry plasmid、PBS were respectively transfected into the mice as Empty plasmid Group、Blank Group. The concentration of IFN-γ in the serum was detected by ELISA. The expression of SOCS1、PTEN mRNA in the liver was detected by qPCR at 30d post-transfectioned. The Western blot was performed to detect the changes in SOCS1、PTEN、HBX in the liver tissue at 30 d post-transfectioned. The results were analyzed with Student’s t-test, or one-way analysis of variance and the least significant difference test. Results: the colony PCR、double enzyme digestion and sequencing verified that the gene was inserted into the pmR-mCherry vector. Compared with Blank Group, the expression of miR-155 in the Experimental Group was significantly increased(t = 8.90, P < 0.01); the concentration of IFN-γ in the Experimental Group was significantly increased(F = 26.58, P < 0.01); the mRNA(F_{SOCS1 mRNA} = 19.72, P < 0.01; F_{PTEN mRNA} = 7.38, P < 0.05) and protein(F_SOCS1 = 50.30, P < 0.01; F_PTEN = 129.00, P < 0.01) expression of COCS1、PTEN was significantly decreased in the Experimental group and the protein of HBX was also significantly(F_HBX = 77.97, P < 0.01). Conclusion The pmR-155 eukaryotic overexpression vector is successfully constructed, this recombinant vector can express miR-155 stably; miR-155 can down-regulate cocs1、PTEN gene expression and up-regulate the expression of IFN-γ, it can inhibit the replication of HBV and a potential targets to treating hepatocellular carcinoma.

Objective To evaluate the effect of intra-aortic balloon pumping(IABP)in treating serious coronary heart disease. MethodsA retrospective analysis was performed on 19 patients who suffered from serious coronary heart disease and accepted IABP therapy,the differences of mean arterial pressure before and after treatment were compared.In order to compare the in-hospital mortality,the patients were divided into 2 groups:6 of 19 patients accepted single IABP therapy,13 of 19 patients attempted IABP and revascularization(thrombolytic/percutaneous coronary intervention/coronary artery bypass graft)therapy. ResultsBedside success rate of IABP operation was 100%without complication.Effective rate was 89.5%(17/19),2 patients who were irreversible phase of cardiogenic shock,were an ineffective treatment.The patient's mean arterial pressure increased from(52.1 ± 18.4)mm Hg to(78.3 20.8)mm Hg after using IA BP for 30 minutes(P＜0.01).The in-hospital mortality was significantly lower in patients received revascularization therapy in addition to IABP compared with patients who had IABP support alone 7.7% vs 83.3%(P＜0.01). ConclusionIABP in treating serious coronary heart disease was safe and effective.IABP treatment before irreversible phase of shock and revascularization therapy following IABP are the key to decrease in-hospital mortality.