| INTRODUCTIONUreteropelvic junction obstruction is one of the most common renal diseases. It can result in severe hydronephrosis and renal failure. Its pathogenesis can be divided into 3 parts: intrinsic, extrinsic, and secondary. We mainly discuss the kind of UPJO caused by intrinsic reason.The pathophysiological process of UPJO is still unknown. People found that there was some confusion of the mechanism of peristaltic activity of renal pelvis. In the view of histology, mass of longitudinal smooth muscle bundles took the place of normal helical smooth muscle bundles, and the renal pelvic smooth muscle became hypertrophic. People didnt know if the change was primary or not, but they believed that it was not the single reason of that UPJO developed the confusion of the peristalsis of renal pelvis and lead to hydronephrosis.There are many other facts that affect the peristalsis of renal pelvis besides we refer to above paragraphs. Myogenic and neurogenic facts control the peristalsis of renal pelvis. Now we know that there is beta - adrenergic receptors that may have a role in blockade of the peristalsis of renal pelvis in UPJ. With the use of beta - AR agonist, people can decrease the rate of the peristalsis of smooth muscles in renal pelvis and restrain the capability of the contraction of smooth muscles. To date, no one has reported the distribution of beta 3 AR in UPJ. In the present study we detected the quantity of beta AR ( betal, beta2, beta3 AR) and compared the difference of that in normal UPJ and in UPJO ones.MATERIALS AND METHODS1. Tissue samplesThis study used data from 47 patients (31 men and 16 women) who underwent surgery for renal cell carcinoma (15 ) or hydronephrosis ( 32). The mean age of the patients was 45. 36years ( range 16 to 73 ). The number of left UPJ is 30,and of right is 17. Among the 32 hydronephrosis patients, 20 were primary hydronephrosis patients caused by congenital UPJO, 12 were caused by other reasons.All the specimens were suitable for immunohistochemical analysis and western blotting in all cases. The classification of the pathogenesis was made by the method motioned by < Campbell's Urology > , the histological diagnosis was carefully examined by 2 experienced pathologists.2. ReagentsThe primary antibodies used in this study were anti - betal AR rabbit poly-clonal antibody at 1:100 dilution (Santa Cruz Inc. , USA), anti - beta2 AR rabbit polyclonal antibody at 1:100 dilution (Santa Cruz Inc. , USA) , and anti -beta3 AR goat polyclonal antibody ( (Santa Cruz Inc. , USA). The Ultra -sensitive S - P kt and DAB agent kit were bought from Peking Zhongshan Biological Company. The goat - anti - rabbit second antibody was bought from Huamei Biological Company.3. Methods3. 1 Immunohistochemical analysis; The standard - labeled streptavidin -biotin technique was used for immunohistochemical staining. In brief, 5 - um sections were cut. The sections were incubated overnight at 4 centigrade with anti - betal, beta2, beta3 polyclonal antibody. The incubations were followed by sequential 30 - minute incubations with biotinylated - linked antibody and peroxidase - labeled streptavidin. Beta AR were carried out in 5 fields at 100, 200,400 magnification. For evaluation of beta AR immunostainings in smooth muscle cells, cases in which at least 10% of cells were immunoreactive were defined as being positive.3. 2 Western blottingRapidly homogenize tissues (1 - 2g) in 5 volumes of lysis buffer (50mM Tris - HCL pH 7.4, 0.1% SDS, 1% Triton -100, 1 mM EDTA PH8.0, Apro-tinin 12ul/10ml, PMSF 60ul/10ml) centrifuge the homogenate ( 10,000rpm,4C ) for 30 minutes to pellet insoluble material. (1) Electrophoresis (2) transfer proteins from gel to membrane (3) blocking (4) incubation with primary antibody (5) enzyme conjugate incubation (6) substrate incubation.4. Statistical analysisAll statistical analysis was performed using commercially available software (SPSS 10. 0). Both the T - Test and one - way - ANOVA were used to assess the difference of t...
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