ABSTRACT
Objective To investigate the efficiency of target gene transfection of the heart and liver after tail vein or intramyocardial injection of adenovirus vector (GFP-Ad).Methods GFP-AD was constructed at first.A total of 20 male 8-week old C57BL/6 mice were randomly and equally divided into tail vein injection of GFP-AD group and intramyocardial injection of GFP-AD group.The mRNA levels of GFP in the heart and liver tissues were detected by Q-PCR at different time points.Fluorescence microscopy was performed to visualize the expression of GFP fluorescence.Results Compared with the tail vein injection group, the GFP mRNA level in mouse heart tissue was apparently higher in the intramyocardial injection group.In both groups, the GFP mRNA levels in liver tissue were significantly increased compared with that in the heart tissue.In the tail vein injection group, the GFP mRNA level in liver tissue reached a peak on day 7;but in the intramyocardial injection group, the mRNA level of GFP in liver tissue reached apeak on day 3.We also observed the same trend of GFP fluorescence expression in the tail vein injection group compared with that in the intramyocardial injection group.Conclusions Intramyocardial injection of adenovirus vector is suitable to achieve a higher transfection efficiency in mouse heart tissue compared with the tail vein injection method.Although both injection methods are suitable for transfection of mouse liver, the tail vein injection method is preferential for it is simple and less invasive.
ABSTRACT
Objective To establish a porcine allogeneic left lung orthotopic transplantation model to closely simu-late human lung transplantation.Methods Twelve Huanjiang mini-pigs were used as donors and 12 Bama mini-pigs as recipients.The left lung orthotopic transplantation was completed by the left fourth intercostal thoracotomy.At 1 h, 2 h, 4 h, 6 h, 12 h after transplantation, the left and right pulmonary artery pressure were measured, the left and right pulmonary vein blood gas was analyzed, and samples of the left and right lung tissues were taken to determine the water content and for pathological examination.Results All animals survived, and the transplanted pulmonary vein blood PaO2/FiO2 and PAP were rised along with the prolonged postoperative time, compared with those of the recipient normal lung showing a signifi-cant difference (P<0.05).With the pass of time, there were increasing edema, inflammatory cell infiltration, RBC ooze, thickening of alveolar wall in the transplanted lung tissue, and some alveolar lumen occlusion and lung tissue consolidation. The water content of the transplanted lung tissue was increased significantly compared with that in the recipient lung tissue ( P<0.05 ) .Conclusions The established method in this study provides an ideal animal model for research on lung transplantation ischemia-reperfusion injury and immune rejection mechanism.
ABSTRACT
Objective To modify the techniques for establishment of an abdominal working heart transplantation model in rats and to sum up the key factors to success.Methods A total of 180 12-week old Brown Norway rats ( donor) and Lewis rats ( recipient) were used in this study:50 BN rats and 50 Lewis rats for pilot experiment, and 40 BN rats and 50 Lewis rats for the formal experiment.The rat model of working heart heterotopic transplantation was adopted and estab-lished by Wiedemann’ s mode.We transplanted the heart from BN rats to Lewis rats and analyzed the survival rate, causes of death and histological changes of the heart ( HE staining) in this experiment.Results After exercise and modification, the survival rate was increased to 77.5%, and the mean total duration of operation was 71 ±11 min, and the mean ische-mic time of the donor hearts was 34 ±5 min.Histological examination ( HE staining) of the cardiac allograft showed a mild inflammatory cell infiltration in the graft at 24 h after transplantation, indicating that the model was reliable.Conclusions A variety of factors may affect the final operation success rate in the establishment of this heart transplantation model.A-mong them, the major affecting factors include: healthy animals, donor heart protection, rapid and effective vascular su-ture, and postoperative animal management.
ABSTRACT
Objective To evaluate the anatomic localization and size of acute necrotic myocardium in the ischemic-reperfused rat hearts using 99m TC-Glucarate and microSPECT/CT.Methods The ischemic-reperfused ( IR) rat heart models were established by ligating left anterior descending coronary artery for 60 min.99mTC-Glucarate was intravenously injected into the rats 24 hours after IR operations .Images were acquired 30 min after administration of 99m TC-Glucarate using microSPECT/CT. Anatomic localization and size of acute necrotic myocardium were analyzed with microSPECT/CT imaging , and these results were compared to those determined by triphenyltetrazolium chloride ( TTC ) staining.Results The microSPECT/CT images showed hot spot accumulations of 99mTC-Glucarate in IR hearts (the heart-to-liver ratio was 1.90 ±0.33), not in controls (P <0.05).The anatomic localization of 99mTC-Glucarate-labeled necrotic myocardium were in correspondence with TTC staining results .The hot spot size was related significantly to necrotic myocardial size determined by TTC staining ( R2 =0.964 ) .Conclusions The localization and size of acute necrotic myocardium can be assessed by non-invasive microSPECT/CT imaging with99m Tc-Glucarate.