| Objective1. To establish rat orthotopic left pulmonary allograft transplantation by improving the "cuff-like" vessel anastomosis technique, which will result in a stabler model, a higher operating successful rate and a higher postoperative survival rate ;2. To examine the feasibility of ex vivo adenovirus-mediated gene transfer of human interleukin-10 via the pulmonary vein into rat lung isografts or allografts ,and to investigate the effect of gene transfer on subsequent ischemia-reperfusion injury and acute pulmonary allograft rejection.Methods1. This study improved the traditional cuff-like vessel anastomosis technique in many aspects, including graft retrieval and cuff self-making .The anastomosis of pulmonary artery, vein and bronchus was performed by using the improved cuff technique. When the transplanted lung had been reperfused for 4h, the right hilus of lung was clipped. Just before and 20 min after clipping, systemic PaO2 and PaC02 were determined in blood samples from abdominal aorta. And chest roentgenography, chest CT and pathologic study were conducted 41u , 5 d, 14d after operation, respectively.2. Male SD rats underwent left lung isotransplantation. Isografts were transvascularly transfected 15 minutes after harvest with 5 mL of 5 X 109 PFU adenovec-hlL10 complex (group D, n = 7) or empty adenovec (group C, n = 7) or LPDG (diluent,group C, n = 7). The false operating control is group A (group A, n = 7). All allografts were preserved for 3 hours at 10 degrees C before transplantation and assessed 4 hours after reperfusion. Lung function was assessed by oxygenation (FIO2, 1.0) and wet-to-dry (W/D) weight ratios. GraftMDA,SOD , MPO were measured.Transgene expression of human interleukin 10 was assessed by both reverse ranscriptase-polymerase chain reaction and immunohistochemistry. Graft rat tumor necrosis factor alpha (rTNF-alpha), interferon gamma (IFN-gamma), Fas, Fasl were detected by immunohistochemistry (IHC). Graft rTNF-alpha, IFN-gamma were also measured by ELISA . Pathological morphologic change and apoptosis were also analyzed.3. Male SD rats underwent left lung transplantation with allografts from Wistar rats . Allografts were transvascularly transfected 15 minutes after harvest with 5 mL of LPDG (group I , n = 7) or adenovec (group II, n = 7) or adenovec-hlLlO complex (group III, n = 7) .All allografts were preserved for 3 hours at 10 degrees C before transplantation. In all groups recipients were killed on postoperative day 5. Transgene expression of human interleukin 10 was assessed by means of both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Graft rat interleukin 2, TNF-alpha,IFN-gamma,Fas,Fasl were detected by immunohistochemistry (IHC). Graft interleukin 2, rTNF-alpha, IFN-gamma were also measured by ELISA .Histologic rejection score, allograft gas exchange and apoptosis were also assessed.Results1. In 30 consecutive cases, 27 have been performed successfully by single surgeon.The operation successful rate was 90%, and the survival rate was 89% on the postoperative day 14. The total time of surgical procedure was (58 + 9)min,and the average graft ischemia time was (23 5)min . On 4h and 5 day after operation, chest roentgenography and pathologic study found the typical pathologic change of ischemia-reperfution induced lung injury and acute pulmonary allograft rejection.2. Transgene expression of human interleukin 10 was detected by means of both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Partial pressure of oxygen (PaOi) levels in the hIL-10 group were higher, but W/D ratios was lower than in empty vector and diluent groups (p <0.01). MDA ^ MPO were lower,but SOD was higher than the other groups. rTNF-alpha, INF-gamma , Fas and Fasl were reduced in the tissue of hIL-10 lungs (p < 0.01). Fewer tissue edema ,interstitial inflmmation and apoptosis index were found in h...
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