| Objectives: To ascertain the value and importance of renal graft biopsy and assaying of the level of granzyme B mRNA and perforin mRNA in urine on the diagnosis of renal acute rejection.Methods: Altogether 34 cases of renal transplantation were used in this study. There were 28 male cases, 6 female cases, mean age 36.6 years(range 18-52). fter transplantation, 20 cases had normal BUN (blood urea nitrogen) and Cr(creatinine), 10 cases were clinically diagnosed acute rejection and 4 cases had delayed renal graft function. Renal biopsy (RB) were conducted in 34 cases under the real-time ultrasound guided (26 cases) and CT (8 cases). Urine sample was collected in 34 cases before biopsy (one month before in 30 cases, 1 to 2 months before in 4 cases). The urine sample were measured with the use of a competitive and quantitative polymerase-chain-reaction assay for granzyme B-5-mRNA and perform mRNA. Data were analysed by using the SPLM statistical software, mRNA levels were log-transformed.Results: Biopsy was conducted in 34 cases, altogether 37 times of puncture were done with 3 cases had repeated puncture. The success rate of puncture for ultrasound guided was 89%, for CT was 100%. There were 2 cases (0.06%) had slight hematuria and disappeared 2 days later. There was no other complication in these cases. Between the group of ultrasound (n=29) and CT (n=8) guided biopsy, the number of renal glomeruli was 13.9 ?.2vs. 11.1?.7,(/}>0.05); number of arteries was 2.6?.7 vs. 2.1 ?1.1, (/)>0.05); unsable biopsed sample 3 (10.3%) vs 0; duration of puncture 1.0 min?.5 min vs 5.0 min.?.0 min. According to the classification of Banff in 1997, normal renal function (n=20), there were 18 cases were normal, 2 cases were in critical status; clinically diagnosed acute rejection (n=10), there were 7 cases with acute rejection, among them 2 cases IA, 3 cases IIA, 1 cases IIB and 1 cases III. There were 2 cases with cyclosporine A intoxitation and 1 case with acute tubular necrosis; among the group of delayed renal graft function (n=4). there were 2 cases with acute tubular necrosis,! cases with cyclosporine A intoxitation and 1 case with acute tubular necrosis accompanied with acute renal.rejection. Compared with pathological diagnosis, the diagnostic accordance was 73.5%. The level of granzyme B mRNA and perform mRNA in urine was higher in patients with acute rejection than in patients without acute renal rejection; the level of perform mRNA in patients with acute rejection was 1.2?.4fg/ug of total RNA (/><0.001) and in patients without acute rejection was -0.6?.3fg/ng of total RNA (/J<0.001) . In 10 cases with acute rejection, there were 3 cases had the-6-urine sample 2 months later, and the level of granzyme B mRNA and perforin mRNA in urine had no difference with the other 7 cases(perforin,1.2?.3fg/jj.g vs. 1.3?.4 fg/ug of total RNA, />=0.711, granzyme B, 1.2?.4fg/n.g vs.l.5?.3fg/ug of total RNA,P=0.22), there were 4 cases with acute rejection had IA pathological grade or even lower, the level of perforin mRNA was 1.4?.3fg/u.g of total RNA, and the other 6 cases had pathological grade II or HI, the level of perforin mRNA was 0.9?.5fg/ug of total RNA (P=0.114), and the levels of granzyme B mRNA in grade IA,II and III were 1.3?.4fg/u?and 1.4?.3fg/ug(JP=0.699). When the level of perforin mRNA reached 0.9 fg/ug, it had the largest sensitivity and specificity for diagnosing acute rejection, they were 85% and 83% respectively; when the level of granzyme B mRNA reached 0.4 fg/ug, sensitivity and specificity were 81% and 78%, respectively (/*<0.001). In three cases had DGF caused by non-immune factors, the level of granzyme B mRNA and perforin mRNA in urine was lower than in patients with acute renal rejection(perforin, -0.9?.4fg/ug vs.l.2?.4fg/ug of total RNA, P<0.001; granzyme B, -0.5?.4fg/ug vs.l.l?.4fg/ug of total RNA, /><0.001). In the only one case who had DGF caused by ATN with acute rejection, the level of granzyme B mRNA and perforin mRNA was 1.1 fg/ug and 1.0 fg/ug respectively.Conclus... |
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