Objective To construct a mouse model for real-time,noninvasive and specific monitoring of inflammation activation in hepatic tissues.Methods An inflammation reporter gene was targeted to the liver by hydrodynamic gene delivery technology.Bioluminescence imaging was used to detect the firefly luciferase(Fluc) expression in the mouse liver after inflammatory stimulation.Besides,the relevance between the light intensity and inflammation level was also intensively investigated.Results pIL-6-Fluc was successfully delivered to the liver.The hydrodynamic gene delivery could cause a transient liver injury that could return normal in 5 to 7 days.The expression of pIL-6-Fluc could be induced by lipopolysaccharides(LPS) treatment with an about (46.80±13.35) fold increase at the peak value,which was significantly higher than that detected by ELISA [(4.09±0.96)fold].Conclusion An inflammation reporter mouse model is constructed in this study by hydrodynamic gene transfection,allowing noninvasive monitoring of inflammation activation specifically in hepatic tissues.The reporter model is capable of monitoring inflammation activation with a sensitivity higher than that of ELISA.

Objective To investigate the expression and function of transforming growth factor (TGF)-β1 and Smad2 in liver fibrosis of biliary atresia (BA). Methods Liver biopsy specimens were collected from autopsy (normal group, n=5), congenital biliary dilatation (CBD group, n=10), BA patients underwent Kasai procedure (early hepatic fibrosis group, n=19) and liver transplantation (transplantation group, n=11). The first three groups were collected from January 2010 to July 2014 in Tianjin Children’s Hospital, and the last group was collected from January 2013 to January 2014 in Tianjin First Central Hospital. The hematoxylin and eosin (HE) stain were used to observe the degree of liver fibrosis of four groups. Immunohistochemistry (IHC) was used to observe expressions of TGF-β1 and Smad2 in liver tissues of these samples. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to test the quantitative mRNA of TGF-β1 and Smad2 in these samples. Results Results of HE showed that no fibrosis in autopsy group, mild fiber cell hyperplasia in CBD group, severe fibrosis in Kasai group and significant pseudolobule in transplantation group. Results of IHC showed that TGF-β1 was expressed in the cytoplasm of hepatocytes, bile duct cells, lymphocytes and neutrophils. The average optical density of TGF-β1 was the highest in Kasai group compared with that of other three groups (P < 0.05). There was no significant difference in Smad2 expression in cytoplasm of hepatocytes, bile duct cells and lymphocytes between four groups (P>0.05). Results of qRT-PCR showed that both TGF-β1 mRNA and Smad2 mRNA were the highest in early hepatic fibrosis group than those of CBD group and transplantation group (P<0.017). Conclusion In early stage of BA, TGF-β1 and Smad2 promote liver fibrosis until the formation of P-P,P-C desmosome structure. However, with BA fibrosis becomes more serious, the pro-fibrogenic function of TGF-β1 and Smad2 becomes less.

Purpose To study the values of liver histopathological assessment grading score in differential diagnosis between biliary atresia ( BA) and infantile hepatitis ( IHS) . Methods Thirty four cases of BA and sixteen cases of IHS were analyzed retrospectively, which were diagnosed by biopsy. A hepatic histopathological assessment grading score was developed. This consisted of eight features such as cholestasis, hepatocellular damage, bile duct proliferation, portal edema, portal inflammation, portal fibrosis, extramedullary hemopoiesis and multinucleated giant hepatocytes. The total scores were 24 points. All the cases were assessed one by one. Results The total scores of BA were significantly higher than that of IHS (P<0. 001). The frequencies of bile duct proliferation, portal fibrosis and portal edema were significantly higher in BA than that in IHS group, while the frequency of multinucleated giant hepatocytes was significantly higher in IHS than that in BA group. Conclusions This scoring system is helpful in differentiating BA from IHS.

Objective To establish a stable HCV full-length genome replication cell model which is labeled with reporter gene and easyly to quantify intracellular HCV proteins and RNA level. Methodsneo gene was inserted into Luc-JC1 to make Luc-JC construct. Luc-JC RNA was obtained by in vitro transcription and then delivered into Huh7 cells by transfection. G418-resistant clones of Huh7 cells were obtained by selection. Clones of HCV full-length genome replication cell were confirmed by luciferase activity assay, Western blot and cleaveage of eYFP-MAVS by HCV NS3/4A protease. Then, HCV replication cell colonies were treated by different dose IFN-α in order to observe the change of luciferase activity, HCV protein and RNA level. Results At 3-4 weeks post-transfection, visible colonies were selected and stained by crystal violet. Luciferase activity and HCV NS3, NS5A protein were detected by luciferase activity assay and Western blot, respectively. Subcellular localization of eYFP-MAVS transferred from mitochondria to cytoplasms by cleavage of NS3/4A protease in cell colonies. Luciferase activity, HCV protein and RNA diminished obviously after IFN-α treatment. Conclusion A stable HCV full-length genome replication cell model labeled by reporter gene was successfully established and reporter activity can be used to indicate level of HCV proteins and RNA in cells. This cell model is a useful tool for the study on HCV pathogenesis and the screening of antiviral drugs.

Purpose　The aim is to investigate the effects of BSP on immunological function in normal and immunosuppressed mices.Methods　Thymus and spleen indexes, peripheral blood WBC number,the lymphocyte proliferation response, IL-2 production and serum and splenocyte hemolysin contents were measured after intraperitoneal injection of BSP in normal and immunosuppressed mices.Results　(1)BSP 100 mg/（kg*d）×10d significantly increased the thymus and spleen indexes and peripheral blood WBC number in immunosuppressed mice.The thymus and spleen indexes in normal mice was also increased. In addition,BSP markedly improved T,B lymphocyte proliferation responses and IL-2 production in normal and immunosupressed mices.(2) BSP improved the serum and splenocyte hemolysin contents in normal and immunosuppressed mice. Conclusion　It was suggested that BSP was a kind of immunomodulator, and could improve the immunological function of normal and immunosuppressed mices.

Objective:To investigate the presence of SENV infection among patients in China,and analyze partial nucleotide sequence of SENV isolated from a patient with non A-G hepatitis.Methods:A nested polymerase chain reaction (PCR) assay with primers from ORF1 of SENV genome was established to detect SENV DNA.The PCR product was cloned and sequenced.Results:SENV DNA was positive in 2 of 7 patients with non A-G hepatitis and TTV negative.Partial gene of a SENV isolate was compared with the corresponding region of SENV isolate(AX025730)from Italy and was found that the nucleotide homology was 90%.Conclusions:The results of this study confirmed the presence of SENV infection in China.The development of a PCR assay for SENV DNA detection and the cloning,sequencing of the SENV isolate have important implication for the diagnosis and epidemiological investigation on SENV infection.

Objective: To investigate the effects of BSP on lymphocyte function and cytokines production in normal and immunosuppressed mice.Methods: The lymphocyte proliferation response, T lymphocyte subpopulations and IL 2 production were measured after intraperitoneal injection of BSP in normal and immunosuppressed mice, and the activity of IL 1 and TNF secreted by mouse peritoneal macrophages were detected after BSP supplementation in vitro.Results: (1)BSP〔(100 mg/(kg?d)?10 d)〕markedly improved T,B lymphocyte proliferation responses and IL 2 production in normal and immunosuppressed mice, and significantly increased the number of L 3T + 4 cells in thymus and spleen of immunosuppressed mice, and the ratio of L 3T + 4/Lyt 2 + cells was also increased.(2)BSP distinctly enhanced the activity of IL 1 and TNF production by mouse peritoneal macrophages in vitro.Conclusion: BSP can improve the immunological function of normal and immunosuppressed mice.