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Objective To construct the micro-invasive immune rejection monitoring methods with peripher-al blood mononuclear cell gene expression detection and evaluate the clinic rejection estimation value. Methods The SYBR Green Ⅰ was used as fluorescent dye and the GAPDH as house keeping gene control in the quantitatiun RT-PCR technique to observe the 16 immune rejection relative genes expression features after heart transplantation. results were also compared with that of the normal people. Results The 16 immune rejection relative genes expres-sion were no different between normal people and the transplantation recipients before surgery (P>0.05). After heart transplantation the expression of ITGA4, FKB, ILI R-2 up regulated and the level of PF4、ITGAM、TGFβ1、 RHOU down regulated. The results were similar with the clinic observation that the immune rejection often occurs in the first 3 months after heart transplantation. It implied that these 7 genes may play an important role in the acute im-mune rejection after transplantation. Conclusion The real time quantitation RT-PCR methods were constructed suc-cessfully to detect the multiple immune relative genes expression and is of chnic applicable.
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Differential gene expression profiles in Balb/cJ mouse model of acute hepatic failure in- fected with MHV-3 virus intervened by anti-hepatic failure compound (AHFC) and the changes of cytokines regulated by genes were investigated. The Balb/cj mice were divided into AHFC-intervened group and control group randomly. Acute hepatic failure model of Balb/cJ mice infected with MHV-3 virus was established. The survival rate in the two groups was observed. It was found that the survival rate in the AHFC-intervened group and control group was 90% and 50% re- spectively 48 h after intrapefitoneal injection of MHV-3 (P<0.05). Before and after the experiment, the cytokines in peripheral blood of the survival mice were determined, and RNA was extracted from survival mouse liver tissue for the analysis of the differential gene expression by a 36 kb mouse oli- gonuleotide DNA array. In all the genes of microarray there were 332 genes expressed differently in the two groups, in which 234 genes were up-regulated and 78 genes down-regulated. Through clustering analysis, the differential expression of immune related genes, including TNF receptor superfamily, Kctd9, Bcl-2, Fg12, IL-8, IL-6, IFN-γ, TNF-α etc. might be related with the curative effectiveness of AHFC. It was suggested that AHFC can balance the immune state of mouse model of acute hepatic failure infected with MHV-3 virus mainly through regulating the expression of immune related genes, decrease the immune damage and inhibit liver cell apoptosis of mouse acute hepatic failure model obviously so as to increase the survival rate of mouse models of acute hepatic failure.
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A model of chronic hypoxic pulmonary hypertension (HPH) was reconstructed in rats exposed to hypoxia with normal pressure for 14 d. Chronic HPH was prevented by giving Ligustrazine (LTZ) in an intraperitoneal dose of 80 mg ?kg-1 wt. twice daily. The mean pulmonary arterial pressure (mPAP), plasma and pulmonary small arterial cGMP level, and mRNA expression of nitric oxide synthase (NOS) gene in lung tissues were studied. The results showed that the mPAP was significantly higher, but plasma and pulmonary small arterial cGMP level as well as mRNA expression of NOS gene in lung tissues were markedly lower in chronic hypoxic rats than those in control animals. LTZcould reverse those changes of above indexes in hypoxic rats, but did not affect those indexes in normal rats. There was a strong negative correlation between cGMP level and mPAP. It is suggested that the lower mRNA expression of NOS gene in hypoxic rats may be one of pathogenetic mechanisms in chronic HPH, and LTZ can increase the mRNA exprssion of NOS gene and the production of nitric oxide in hypoxic rats, which may be an important mechanism in LTZ preventing chronic HPH.