Objective To study hepatotoxicity and superoxide dismutase(SOD) acitvity produced by mixture of Nickel Sulfate,Potassium Dichromate and Cobalt Chloride.Mehtods Hepatotoxicity and superoxide dismutase(SOD)activity of mixture of Nickel Sulfate,Potassium Dichromate and Cobalt Chloride were detected by ip in mice.Results The liver weights were increased consistently during three weeds treatment.It was showed in histology study that hepatocytes swelling,necrosis and congestion wigh blood in venae interlobares hepatis.The SOD activity of liver and blood decreased after one week treatment but increased the following two weeks treatment.Conclusions Mixture of Nickel Sulfate,Potassium Dichromate and Cobalt Chloride had obvious hepatotoxicity and a certain lipid peroxiodation effect in vivo.

Objective To observe the change of composition of PL and content of protein in PS after anatoxin damnify lung.Methods The rats lung injury models were made by intratracheally instilling anatoxin(0.1mg/kg,0.4mg/kg).24 rats were divided into four groups:12hour group,2day group,3day group,5day group.Pr of each group were examined by electron microscope,content of PL,composition of PL and content of protein of each group wete determinde respectively.Results Rats lungs in experimental guoups were found that PS lost continuously,dropped in the pulmonary alveolies.12hour group was more apparent.Content of protein in PS was the highest in 12hour group,Content of PL in PS go add,12hour group was more apparent.Conclusion Morphologic change and alternation of quality and quantity of PS after anatoxin-induced pulmonary injures specifically reflect the activity of lung danmify.Measuring content of PL and PI is sign of simple and feasible method when lungs of the rats are injured early by bleomycin.

Objective To study the adverse effect of amatoxins on DNA in the histiocytes of rats and the antagonism of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA). Methods SD rats were randomly divided into three groups, the number of males and females was the same. The rats in the experimental groups were respectively treated with amatoxins(0.1 mg/kg bw, i.p.) and amatoxins added EPA, DHA (both were 10 mg/kg bw, i.p.) for 10 days. Single cell gel-electrophoresis was employed to test the damage of DNA in the histiocytes. Results In the group treated with amatoxins, DNA damage in the peripheral lymphocyte, hepatocytes, lung cells ,brain cells was more seriously compared with the control, but no significant DNA damage was seen in the group treated with amatoxins added EPA, DHA. Conclusion Amatoxins may induce DNA damage and eicosapentaenoic acid, docosahexaenoic acid have an antagonism to the adverse effect of amatoxins.

For rapid and accurate screening of recombinant modified vaccinia virus Ankara (rMVA) that satisfied the quality standards of clinical trials, a novel shuttle vector that can delete the marker gene automatically during virus propagation was construted: pZL-EGFP. To construct the pZL-EGFP, the original shuttle vector pSC11 was modified by replacing the LacZ marker gene with enhanced green fluorescent protein (EGFP) and then inserting homologous sequences of TKL into the flank regions of EGFP. Baby hamster kidney (BHK)-21 cells were cotransfected with pZL-EGFP and MVA, and underwent ten passages and one plaque screening to obtain the EGFP-free rMVA carrying the exogenous gene. Resulting rMVA was tested by polymerase chain reaction and western blotting to verify pZL-EGFP function. A novel shuttle vector pZL-EGFP containing an EGFP marker gene which could be deleted automatically was constructed. This gene deletion had no effect on the activities of rMVA, and the exogenous gene could be expressed stably. These results suggest that rMVA can be packaged efficiently by homologous recombination between pZL-EGFP and MVA in BHK-21 cells, and that the carried EGFP gene can be removed automatically by intramolecular homologous recombination during virus passage. Meanwhile, the gene deletion had no influence on the activities of rMVA and the expression of exogenous target gene. This study lays a solid foundation for the future research.
Animals ; Biomarkers ; Cricetinae ; Epithelial Cells ; virology ; Gene Deletion ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombination, Genetic ; Vaccinia ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication

Objective Constructing the dendritic cell (DC) vaccine loading HPV16L1 antigen,and evaluating the specific humoral and cellular immune responses induced by DC-L1.Methods The mature dendritic cells (DCs) were induced in vitro from bone marrow precursors by co-culturing with cytokines cocktail,and then transduced with a recombinant adenovirus harboring the HPV16L1 gene to obtain the DC-L1 vaccine.C57BL/6 mouse were immunized intramuscularly by single dose of DC-L1 and recombinant adenovirus vaccine Adv-HPV16L1 respectively.The specific humoral and cellular immune responses were determined by quantitative ELISA and ELISPOT at different time points.Results DC-L1 cells exhibited typical morphological features of dendritic cells,and the expression of HPV16L1 can be detected by western blot.Both DC-L1 and Adv-HPV16L1 can induce specific cellular immune response,while with distinct kinetics.Single dose of Adv-HPV16L1 induced responses immediately,and attained the peak level at the third week after immunization,then diminished gradually.While responses induced by DC-L1 elevated slowly,and eventually exceeded that of recombinant adenovirus group at the fifth week of postimmunization.As to humoral immunity,both DC-L1 and Adv-HPV16L1 vaccines can induce specific humoral immune reponses,and with similar kinetics.Conclusion The DC-L1 vaccine was constructed successfully.The DC-L1 vaccine can induce high levels of HPV16L1 specific humoral and cellular immune response.

Objective: To construct a recombinant double gene co-expressing plasmid of HIV-1 broadly neutralizing antibody and to detect its expression in 293T cell line. Methods: HIV-1 neutralizing antibody 2G12 variable region of light chain (VL) and heavy chain (VH) was synthesized, and ligated with vectors containing human IgG constant regions of light and heavy chain to construct a complete 2G12 light and heavy Chain. The VL and VH of 2G12 and IRES were cloned into eukaryotic expression vector pVR by PCR amplification and then the recombinant plasmid was transfected into 293T cells. Expression of the antibody in cell supernatant was detected by ELISA. Binding and neutralizing activity of the cell supernatant were tested by ELISA and micro-neutralization assays. Results: The recombinant double gene eukaryotic expression vector which can express human IgG was constructed successfully. The expression level of the supernatant was 6.43 μg/ml, and the antibody retained the binding and neutralizing activity. Conclusions The constructed vector can express the antibody with binding and neutralizing activity, this study provides a good platform for the expression of human HIV neutralizing antibody in the eukaryotic expression vector.