Objective To construct human EpCAM eukaryotic recombinant plasmid ,and to study the expression of EpCAM pro‐tein in HepG2 cells .Methods The recombinant plasmid ,named pcDNA3 .1(+ )‐EpCAMwas constructed by cloning EpCAM gene into eukaryotic vector pcDNA3 .1(+ ) .Then HepG2 cells were transfected with EpCAMrecombinant plasmid .The eukaryotic ex‐pression of EpCAM protein was verified by immunofluorescence ,Western Blotting and flow cytometry .Results The construction of EpCAM recombinant eukaryotic plasmid was identified by restriction enzyme digestion .The results of immunofluorescence , Western Blotting ,and flow cytometry consistently indicated that EpCAM protein were successfully expressed in HepG2 cells .Con‐clusion Human EpCAM eukaryotic vector is constructed and EpCAM protein could be expressed well in HepG2 cells ,which laid a foundation for further research on the function ofhepatocarcinoma cells highly expressing EpCAM .

Objective To explore the humoral and cellular immune responses of a vaccine of S fragments from a new type reovirus R4 strain in mice.Methods Four recombinant plasmids were constructed by respectively cloning S 1, S2, S3,S4 genes into pcDNA3.1+, and mice were intramuscularly immunized with the recombinant plasmids in a dose of 100 μg/mouse.Control vector pcDNA3.1 +and phosphate buffered saline (PBS) were used as negative controls.The spe-cific antibody level and IgG subclass (IgG1, IgG2a, and IgG2b) were detected by ELISA, and cellular immune responses to R4 were assessed using an interferon ( IFN)-γELISpot assay .Results All recombinant plasmids induced significantly higher levels of anti-R4 IgG compared with that of the controls (pcDNA3.1 +and PBS), and the titers were highest in the mice immunized with S1 and S3.On the other hand, S1 gene induced highest IgG2a antibody and the cellular immune re-sponse was best .Conclusions After the mice immunized with S 1 gene recombinant plasmid , this plasmid can initiate both cellular and humoral immune responses in mice .S1 gene recombinant plasmid is a promising vaccine candidate .

Objective:The aims of this study were to compare the efficacy between salvage liver transplantation (sLT) and rehepatectomy (RR) basing on the propensity score matching (PSM), and to explore the prognositc factors of patients with recurrent hepatocellular carcinoma (HCC).Methods:124 patients with intrahepatic recurrence after hepatectomy in our center from January 2012 to August 2018 were divided into sLT group( n=46) and RR group( n=78). 34 patients were selected for data analysis base on 1∶1 propensity score matching (PSM). The advantages and disadvantages of the two surgical methods and the prognostic factors of the patients were discussed by comparing the basic clinical data, OS and DFS of the two groups before and after matching. Results:Before matching, the pre-operative serum total bilirubin, the occurrence of multiple tumors, the proportion of preoperative TACE and the proportion of patients within Milan criteria of the sLT group were higher than those of the RR group, and the maximum tumor diameter of the sLT group was shorter than that of the RR group ( P<0.05); the time of operation, the amount of intra-operative blood loss, the positive of MVI and the proportion of postoperative Clavien grade Ⅲ and above of RR group were higher than those of RR group ( P<0.05). After matching, the operation time, intra-operative blood loss and the proportion of postoperative complications of sLT group were higher than those of RR group; there was nosignificant difference between 1-, 3- and 5- years OS of sLT group and RR group( P>0.05), the 1-, 3- and 5-years DFS of sLT group were better than those of RR group( P<0.05); AFP≥100 μg/L was the independent risk factor of OS, and the type of operation and AFP≥100 μg/L were independent risk factors of DFS. Conclusions:The pre-operative condition of sLT group was more severe, and the operation time, intra-operative blood loss and post-operative severe complication rate of sLT group were higher than those of RR group, and the DFS time of sLT group was longer than that of RR group, but there was no significant difference in OS between the two groups.

Objective:To explore the role of arginine metabolism in the inflammatory response to influenza A virus (FluA) infection.Methods:Eighteen-month-old mice were infected with FluA via nasal drip, with samples collected on the 6th day post-infection. The concentration of cytokines was determined by the Luminex multifactor assay, while the metabolites in bronchoalveolar lavage fluid (BALF) were analyzed using targeted metabolomic method. Correlation analysis was employed to examine the correlation between cytokines and metabolites. Macrophages were infected with FluA at a multiplicity of infection (MOI) of 1 and cultured with different concentrations of arginine for 24 h. The mRNA and protein expression levels of interleukin (IL)-1β, IL-6, tumor necrosis factor α (TNF-α) and IL-10 were detected by real time fluorescence quantitative RT-PCR (qRT-PCR) and Cytometric Bead Array (CBA).Results:In comparison to the control group, the levels of surfactant protein D (SP-D), TNF-α, monocyte chemotactic protein-1 (MCP-1), IL-1α, IL-6, IL-10, recombinant S100 calcium binding protein (S100) A9, interferon inducible protein 10 (IP-10), macrophage inflammatory protein-1α (MIP-1α), matrix metalloprotein-9 (MMP-9), and Complement Factor D in BALF of FluA infection exhibited a significant elevation. The concentrations of arginine, aspartate, citrulline, glutamic acid, ornithine, proline, creatine, and sarcosine in arginine metabolism were up-regulated, which was correlated with most of elevated cytokine levels. The supplementation of arginine after FluA infection significantly decreased the levels of IL-1β, IL-6, and TNF-α, but increased the level of IL-10 in macrophages.Conclusions:Arginine reduces the inflammatory response induced by FluA infection in macrophages, suggesting that it may be a potential intervention target for severe pulmonary inflammation following FluA infection.