| Objective:Despite the growing array of immunosuppressive therapies available to transplant recipients, acute allograft rejection is a common event and is a major factor in determining both short-term and long-term outcomes for the transplant recipients. Currently, acute renal allograft rejection is suspected only when the serum creatinine level rises, after the adverse effect of immunological and inflammatory processes on graft function. Surveillance allograft biopsies may prove to be beneficial in predicting rejection, but clinical application is limited by the invasive nature of this procedure. The rapid developments of immunology and molecular biology make it possible to monitor acute rejection by measurement of T-cell activating, which is the center step of acute rejection. So the aims of this research were: (1) to study the anatomical basis of rat kidney transplantation; (2) to establish an easy-performing and high-survival-rate rat kidney transplant model; (3) to establish experimental acute rejection model of kidney transplant and learn its histological features; (4) to quantitative detection intragraft and peripheral blood monocytes immune activation genes such as perform, Fas ligand and CD40 ligand transcripts, analysis the relationship between acute rejection and immune activation genes transcripts, and establish noninvasive diagnostictool for allograft acute rejection. Materials and methods:I. Anatomical basis of rat kidney transplantationThirth-six adult male and female Sprague-Dawley (SD) rats, weighing 250~280g, were used throughout the study. Thirty were used in living anatomical basis research and others casting mould specimen. Arteria abdominalis, vena cava caudalis and their branches were observed. Interrelated vessel sizes were carefully measured in the meantime.II. The experimental model establishment of rat kidney transplantation SD rats were used both as kidney donors and as recipients. Resected donor organs were left kidney, left kidney vessels, arteria abdominalis, vena cava caudalis, left ureter and vesica urinaria. The vena cava caudalis was passed through the lumen of the cuff, and its end was then everted over the cuff and secured in position using a circumferential silk suture. Arterial anastomosis: the whole arteria abdominalis below the renal bifurcation was freed and transected at the midpoint. Continuous sutures were placed in arteria abdominalises of donor and recipient by using extradural catheter as inner supporting tube. When suturing the upper orifice, the extradural catheter ripped into artery through the lower ostium of donor arteria abdominalise. And when suturing the lower orifice, the extradural catheter ripped into artery through arteria mesenterica cranialis. Then arteria mesenterica cranialis was ligated. Venous anastomosis: donor's cuffed vena cava caudalis was inserted into the recipient's renal vein and secured in position using a circumferential silk suture. Urinary tract reconstruction: after ensuring correct orientation of the ureter, the dome of the recipient bladderwas divided. Two stay suturesof 9-0 nylon were placed 180?apart through the recipient bladder and donor bladder patch, which were then held by mosquito clamps. Each side of the bladder was then anastomosed with 4 or 5 interrupted 9-0 sutures.III. Experimental acute rejection model of rat kidney transplantationIn the allograft group Wistar and male SD rats were used as donors and recipients. SD rats served as donors and recipients in the isograft group. The graft specimens were harvested 1, 3, 5 and 7 days postoperatively. They were examined histologically after sectioning and staining with hematoxylin and eosin, Masson's trichrome, periodic acid-Schiff reagent and periodic acidsilver metheramine. Acute rejection was diagnosed according to the Banff 97 classification. This rejection score was prepared semiquantitatively into seven grades from 0 to 6 according to the method stated by Watanabe: 0=normal, l=borderline changes, 2=IA, 3=IB, 4=IIA, 5=IIB, and 6=111 by...
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