| Congenital hydronephrosis is a common urogenital malformation harmful to children's health. Along with the developing of natural course, hydronephrosis turned to be more serious and renal cortex became thinning. Renal function was decreased gradually and tended to be vanishing. Pathological examination showed construction disorder in renal glomerulus and renal tubule, and renal interstitium was replaced by considerable fibrous tissues. Did adhesion molecules (AMs) play an important role in the pathological process that from normal renal tissue to fibrosis? Did intercellular adhesion molecule- 1(ICAM-1) and vascular adhesion molecule- 1(VCAM-1) expression diversity have a relationship with renal pathological changes? Could detection of ICAM-1 and VCAM-1 be a clinical index by which to make judgement about renal pathological lesion? It is still unknown today.At present, most of congenital hydronephrosis need operation. But the operation time and indication were still controversial. The key point was the question about functional evaluation of affected kidney and it could determine whether to keep the affected kidney or not. There are many clinical index and correlation factors to evaluate renal function, such as age, blood urea nitrogen (BUN), serum creatinine (SCr), haemoglobin (Hb) and nuclide nephro-dynamic imaging by using 99mTc-diethylenetriaminepentaacetic acid (99mTc-DTPA) and soon. Our research aimed to find out an ideal index to evaluate renal function to serve clinical work by study these index and factors stated above.In brief, the fundamental part of our research aimed to investigate the renal pathological changes in congenital hydronephrosis, and the chief empirical method was Enzyme-linked immunosorbent assay (ELISA) and Immunohistochemical technique (S-P Two-Step); The clinical part put emphasis on the evaluation of renal function, mainly on nuclide nephro-dynamic imaging by using 99mTc-DTPA. The final purpose was to offer new diagnosis and treatment method in the base of forthcoming knowledge.Materials and Methods1. Fundamental parta) Common MaterialsELISA: 53 children were made definite diagnosis of moderate or severe congenital hydronephrosis by ultrasonic, computed tomography (CT) and intravenous pyelography (IVP).They were 32 boys and 21 girls aged from 28 days to 11 years old. 19 of them were severe hydronephrosis and 34 of them were moderate hydronephrosis. One kidney was affected in 48 children and the others were bilateral. The control group contained 20 normal children; Immunohistochemical technique (S-P Two-Step): The disease groups were the same as ELISA group. The control group contained 9 children, including 4 children with renal duplication, 2 renal trauma and 3 renal embryonal tumors.b) Specimen CollectionELISA: All children were taken peripheral blood on empty stomach, the blood serum was preserved in -80 °C after centrifugation. The urinawas taken from pelvic of affected kidney during operation and kept in -80 °C after centrifugation. Children in control group were taken urina sanguinis and blood before diet; Immunohistochemical technique (S-P Two-Step): All cases were caused by obstruction in pelviureteric junction (PUJ). The renal cortex tissues were taken during operation and the normal tissues taken in control group.c) Equipment and AgentELISA: Multiskan MK3 type of Benchmark plate reader with the wavelength from (450-630)nm was made by Shanghai Instrument Company (Thermol Labsystems).The agent was offered by Invitro Company, France;Immunohistochemical technique (S-P Two-Step): The main agent was PowerVision? Two-Step Histostaining Reagent which bought from Zhongshan Biotechnology Limited Company, Beijing(production of Santa Cruz Biotechnology, Inc. California, U.S.A.).d) Empirical MethodsResearch works were strictly abided by description.e) Statistical MethodsAll of the data were recorded and set up data bank which was performed statistical analysis by SPSS10.0 software. The level of significance was a=0.05. ELISA: One-way analysis of variance (One-Way ANOVA) was utilized firstly, then q test (SNK); Immunohistochemical technique (S-P Two-Step): Positive cell counting in disease and control group were set up data bank and analyzed by t test and correlation analysis.2. Clinical Parta) Common Materials56 children with 68 affected kidneys were made definite diagnosis of moderate or severe congenital hydronephrosis by ultrasonic, CT and IVP. They were 45 boys and 11 girls aged from 26 days to 12 years old. 49 of them were found obstruction in PUJ and 4 in terminal ureter. One kidney was affected in 44 children and the others were bilateral. 28cases were affected in the left, 16 cases in the right. The control group contained 7 normal children; 15 newborn infants with 18 affected kidneys and 12 normal kidneys were made definite diagnosis of moderate or severe neonatal hydronephrosis. They were 12 males and 3 females aged from 5 to 36 days. 11 of them were found obstruction in PUJ, 2 in terminal ureter and 2 with posterior urethral valva. One kidney was affected in 12 children and 3 of them were bilateral. 13 cases were administrated operation, and 9 of them kept follow up.b) Exam MethodsSPECT (GE MILLENNIUM, made in U.S.A.) with double detecting head, low energy and high resolution, parallel aperture collimator was used. "mTc-DTPA was injected transvenously and images were taken simultaneously. In the first phase, 30 images were taken continuously in the speed of every frame 2seconds. 20 images were taken continuously in the speed of every frame 1 minute in the second phase. The last phase (lag phase) was in 1 hour later. Then GFR was determined in Gates regulation by computer. Age, BUN, SCr and Hb were determined simultaneously and they were all performed correlation analysis with GFR.c) Statistical MethodsAll of the data were recorded and set up data bank which was performed statistical analysis by SPSS8.0 software. Data bank was analyzed by Dunnett t test and correlation analysis.Results1. Fundamental parta) ELISASerum level of soluble ICAM-l(sICAM-l) and soluble VCAM-l(sVCAM-l) in moderate or severe congenital hydronephrosis and control group: The serum level of sICAM-1 was (594.83±107.99) ng/ml in control group, (717.39±102.33) ng/ml in moderate hydronephrosis group, (770.45± 122.92) ng/ml in severe hydronephrosis group. The serum levels of sICAM-1 in moderate or severe congenital hydronephrosis group were increased significantly than control group (/)<0.01); The serum level of sVCAM-1 was (350.02±l00.38) ng/ml in control group, (434.16±118.41)ng/ml in moderate hydronephrosis group, (456.11±137.27)ng/ml in severe hydronephrosis group. The serum level of sVCAM-1 showed no significant difference between moderate or severe congenital hydronephrosis group and control group CP>0.05); There was no significant difference between moderate and severe congenital hydronephrosis group (P>0.05) .Urine level of soluble ICAM-l(uICAM-l) and soluble VCAM-l(uVCAM-l) in moderate or severe congenital hydronephrosis and control group: The urine level of uICAM-1 was (9.57±5.24) ng/ml in control group, (10.28±5.82)ng/ml in moderate hydronephrosis group, (10.80±6.45) ng/ml in severe hydronephrosis group; The urine level of uVCAM-1 was (18.50±3.37)ng/ml in control group, (18.55±3.16)ng/ml in moderate hydronephrosis group, (18.61±2.85)ng/ml in severe hydronephrosis group. The urine level of uICAM-1 and uVCAM-1 showedno significant difference between moderate or severe congenital hydronephrosis group and control group (P>0.05) .The correlation analysis between moderate or severe congenital hydronephrosis group and control group: There was no dependablity between sICAM-1 and uICAM-1 (r=-0.172, P> 0.05) ; There was no dependablity between sVCAM-1 and uVCAM-1 (r=0.337,0.299; P> 0.05 ) .b) Immunohistochemical technique (S-P Two-Step)The renal tissue ICAM-1 expression in moderate or severe congenital hydronephrosis group and control group: Normal renal tissue ICAM-1 expression was lightly in glomerulus, endothelial cell in parietal layer of Bowman capsule, renal tubule and interstitium; ICAM-1 expression in moderate or severe congenital hydronephrosis group was increased significantly than control group in regions stated above (PO.01). ICAM-1 expression in glomerulus and Bowman capsule were discovered obviously raising in moderate group. Renal tubule and interstitium also expressed ICAM-1 markedly in severe group and renal construction disorder could be seen.The renal tissue VCAM-1 expression in moderate or severe congenital hydronephrosis group and control group: Normal renal tissue VCAM-1 expression was slender in endothelial cell in parietal layer of Bowman capsule, proximal convoluted tubule; VCAM-1 expression was slightly in glomerulus, Bowman capsule, renal tubule and interstitium; There was no significant difference between moderate or severe congenital hydronephrosis group and control group (P>0.05).The correlation analysis between ICAM-1 and VCAM-1: There was no dependablity between ICAM-1 and VCAM-1 both in disease group and in control group (r =0.34, -0.25; P>0.05) .2. Clinical PartObsevation about renal function in congenital hydronephrosis: Total GFR taken from "mTc-DTPA nuclide nephro-dynamic imaging in control group was (103.57± 18.37)ml/min, relative renal function of individual kidney was (48.86 ±5.16)%; Total GFR in light hydronephrosis group was (98.94± 16.67)ml/min, relative renal function of affected kidney was (45.59 ±9.57)%( P > 0.05); Total GFR in moderate hydronephrosis group was (73.44± 10.50)ml/min, relative renalfunction of affected kidney was (36.84 ±9.43)%( P < 0.05); Total GFR in severe hydronephrosis group was (62.83± 10.36)ml/min, relative renal function of affected kidney (21.35±7.39)%( P < 0.001). It could be seen that compare with normal renal function, the decrease of renal function in moderate and severe hydronephrosis was statistically significant. And there was no significance in renal function decrease in light congenital hydronephrosis.Obsevation about renal function in neonatal hydronephrosis: Relative renal function of individual kidney taken from 99mTc-DTPA nuclide nephro-dynamic imaging in control group was (55.76+11.02)%; GFR of affected kidney in light hydronephrosis was (29.91±6.07)ml/min, relative renal function was(46.15± 9.68)%( P > 0.05); GFR of affected kidney in moderate hydronephrosis was (18.75±5.49)ml/min, relative renal function was (38.91 ±8.70)%( P < 0.05); GFR of affected kidney in severe hydronephrosis was (12.01±4.45)ml/min, relative renal function was (29.23 ±7.37)%( P< 0.01). It could be seen that compared with normal renal function, the decrease of renal function in moderate and severe neonatal hydronephrosis was statistically significant. And there was no significance in renal function decrease in light neonatal hydronephrosis.The correlation analysis between GFR and clinical index in congenital hydronephrosis: Positive correlation was discovered between GFR and Age (/?=0.428 ,P < 0.05); It was also discovered positive correlation between GFR and Hb in children with moderate and severe hydronephrosis (r=0.341,0.635; P <0.05, P<0.01) ; GFR was negitive correlated with BUN only in severe hydronephrosis (r= -0.559,P < 0.05); GFR was negitive correlated significantly with SCr(r=-0.445, P<0.0\).The correlation analysis between GFR and clinical index in neonatal hydronephrosis: It was discovered positive correlation between GFR and Hb in neonate with moderate and severe hydronephrosi (r=0.414,0.667; P<0.05, P< 0.01) ; There was no dependablity between GFR and BUN; GFR was negitive correlated significantly with SCr in neonate with moderate or severe hydronephrosis(r=-0.490, -0.756; P<0.05, P<0.01).
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