2.Cloning, Sequencing and Expression of Trichinella spiralis p49 Gene
Yan WEN ; Weide LAO ; Hong GAO ; Chuansheng ZHANG ; Siguo LIU ; Shaobo GAN
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(06):-
Objective To conduct cloning, sequencing and expression of Trichinella spiralis ES antigen p49 gene. Methods RT-PCR was used to amplify the specific gene fragment from the total RNA of Trichinella spirais larvae. The PCR product was ligated to the T-vector and the recombinant plasmid was verified by sequencing. T-p49 and pGE-4T-3 were treated by both BamHI and XhoI. The ligation reaction was catalyzed by T4 DNA ligase. Results The p49 gene was cloned by using RT-PCR. Sequence analysis showed that the p49 gene obtained was consistent to the p49 sequence reported in the database. The expressed protein was shown as a new band at SDS-PAGE. BLAST analysis demonstrated that this p49 gene was 99% identical to the p49 gene reported and to the 43 kDa secreted glycoprotein gene in the database. Conclusion p49 gene from Trichinella spiralis larvae was cloned, sequenced and expressed.
3.Effect of preconditioning of donor liver by breviscapine on liver graft ischemia/reperfusion injury in rats
Huizhen GAN ; Qian HE ; Qiubao AI ; Yujun ZHANG ; Shaobo ZHANG ; Yijie CHEN ; Liang GE ; Chenghua ZHANG
Chinese Journal of Organ Transplantation 2012;33(1):44-47
ObjectiveTo observe the effect of donor liver pretreated by breviscapine on liver transplantation ischemia/reperfusion injury in rats. Methods SD rats served as liver donors and recipients (n =48 each).The recipients were divided into four groups by random number table.The donors in groups A and C were not pretreated with breviscapine,but those in groups B and D were pretreated with 20 mg/L Breviscapine.The cold ischemia time in donor livers of groups A and B was 30-40 min,and that in groups C and D was 12 h. Clotting function, liver function, serum thrombomodulin,caspase3,and relative activity of NF-kB after liver transplantation were assessed,and the pathological changes and TUNEL apoptosis staining were observed.ResultsThe mortality in groups C and D was 40.0% (8/20) and 29.4% (5/17),respectively (P>0.05).There were no significant changes in coagulation function in all groups after operation. The liver function was improved,pathological lesions were alleviated,and apoptosis rate,serum TM,caspase3 expression and activity of NF-kB in the liver tissues of group D were significantly decreased as compared with group C at 3rd day after operation (P<0.01),but all these parameters in group B had no significant change compared to group A.ConclusionPretreatment of donor livers with breviscapine can reduce the ischemia/reperfusion injury and apoptosis after liver transplantation in rats probably by inhibiting the apoptosis-related pathway and alleviating the damage to the endothelial cells of the liver microcirculation.