Objective To explore the relationship between acute rejection and expression levels of perforin mRNA in peripheral blood and bile after rat liver transplantation so as to look for a kind of noninvasive method to diagnose acute rejection. Methods Rat orthotopic liver transplantation model with biliary extra-drainage was established. They were divided into 4 groups: blank control group (n=20)， isograft group (n=30)， allograft plus cyclosporine A (CsA) group (n=30) and allograft group (n=30). Semi-quantatative RT-PCR was used to measure the expressions of perforin mRNA in bile and peripheral blood and the pathological changes of the graft were observed on postoperative days of 1, 3, 5, 7 and 10. Results Blank control group and isograft group showed no expression of perforin mRNA in peripheral blood. The expression of perforin mRNA of peripheral blood in allograft group could be detected on day 3 after transplantation and it gradually elevated on day 5 and remained in high level during 7-11 days. In allograft plus CsA group perforin mRNA of peripheral blood continuously expressed in a low level, and the level at the same time points was significantly different compared with allograft group (P＜0.05). The expression of perforin mRNA in peripheral blood was analogous with the severity degree of histologic damage. The expression of perforin mRNA can not be found in bile. Conclusion The expression of perforin mRNA in the peripheral blood offers a sensitive and noninvasive means of monitoring acute rejection.
Objective To study the effects of pcDNA3/AFP/TK/Angio fusion gene targeting therapy for human primary liver cancer in nude mice implanted with SMMC-7721. Methods Human liver cancer cell line SMMC-7721 was implanted subcutaneously in nude mice to establish experiment model. Animals bearing liver cancer were randomly divided into five groups: control group， vector group， GCV (ganciclovir) group， pcDNA3/TK/Angio group; pcDNA3/AFP/TK/Angio group. Different plasmids were directly injected into tumors and GCV was intraperitoneally administrated simultaneously according to different groups. The growth of tumors was observed and the pathology was examined as well. Serum AFP level was measured by radioimmunology, the ultrastructural change of tumor cells was studied by using electron microscopy, the expressions of MVD and VEGF were respectively detected with immunohistochemistry and the cell apoptosis in situ was detected by TUNEL. Results The success rate to establish subcutaneous implanted liver cancer model in nude mice was 100%. The tumor volume, serum AFP level, VEGF and MVD expressions of pcDNA3/TK/Angio group and pcDNA3/AFP/TK/Angio group were lower than those in control group, vector group and GCV group (P＜0.05) and more apoptosis cells could be observed. While the tumor volume, serum AFP level, VEGF and MVD expressions of pcDNA3/AFP/TK/Angio group was lower than those in pcDNA3/TK/Angio group (P＜0.05); and apoptosis index was higher than that of the latter (P＜0.05).Conclusion pcDNA3/AFP/TK/Angio fusion gene inhibits the growth of tumor remarkably and becomes a promising new biological agent to treat human primary liver cancer.