The Computer Discriminant On Exfoliated Cells In Bladder Urothelial Cacinoma

ObjectiveThis paper was aimed to quantitatively reveal the chromatic and morphologic characters of the exfoliated cell in bladder urotheliual cacinoma by Pap stain, to find highly specific parameters applied to differentiate the exfoliated cell in bladder urotheliual cacinoma, and to discuss the value of the chromatic and morphologic parameters on discriminate the exfoliated cell in bladder urotheliual carcinoma, and establish the technology of diagnosis discrimination.Methods:107 urine samples were collected from the patients, which inculing 21 patients with urotheliual carcinoma of baldder,44 patients with dysplastic urothelium, and 36 patients with normal uroepithelium. The urotheliual exfoliated cells were divided into 500 baldder urothelial carcinoma exfoliated cells (UCC),439 urothelium dysplastic exfoliated cells (UDC) and 386 uroepithelium normal exfoliated cells (UNC). The urine samples were smeared using conventional method, fixed by 95% alcohol, stained by Pap stain. The images of 3 category cells were taken a photograph in objective lens 40 X of light microscope equipped digital camera and inputted into the computer. Technique of computer image analysis was applied to test their relative parameters. The chromatic parameters included three primary colors of cytoplasm (Rp, Gp, Bp) and their coefficient (rp, gp, bp), three primary colors of nuclear (Rn, Gn, Gn) and their coefficient (rn, gn, bn). The morphologic parameters of cell and nuclear included area (Ac, Ap, An), karyoplasms ratio (Rnp), Major axis (dmaj,c, dmaj,n), Minor axis(dmin,c, dmin,n), average diameter(Dc,Dn),circumference (Cc, Cn),Form factor PE (PEc, PEn), Form factor AR (ARc, ARn), Regular form factor (RFFc, RFFn), sphericity (Spc, Spp), Elliptical eccentricity(EEc, EEp)and Form irregular index (FIIc, FIIn). The values of the parameters were tested with one-way ANOVA The following groups were submitted to do the stepwise discriminant analysis. The discriminant functions of random 1077 above cells (training group,80%) were established to analyze the rate of back discriminant, the discriminant rate of residual 248 exfoliated cells (identification group,20%) in urine and 107 cases.Results:Ⅰ.The test results of chromatic parameters for cytoplasm and nuclear of UCC, UDC and UNC in exfoliated cells of urine1. The UNC group has the highest values of Rp, Gp, Bp, Rn, Gn, Bn, the UDC group takes the second place, and the UCC group has lowest values. The F value is respectively 605.296,428.478,281.703,640.236,549.823,318.834, the difference has statistic significance (P=0.001). The multiple comparison showed the values of Rp, Rn, Gp,Gn, Bp, Bn in UCC and UDC were lower than those in UNC, and the values of Rp,Rn,Gp,Gn,Bp,Bn in UCC were lower than those in UDC(P<0.05).2. The UNC group has the highest values of rp, rn, the UDC group takes the second place, and the UCC group has lowest values; The UCC group has the highest values of gp, bp, gn, bn, the UDC group takes the second place, and the UNC group has lowest values. The F value of rp, gp, bp, rn, gn, bn is respectively 304.234,69.621, 177.331,274.429,11.716,204.767, the difference has statistic significance (P=0.001). The multiple comparison showed the values of gp, bp, gn, bn were higher and rp, rn were lower in UCC and UDC than those in UNC, and the values of gp, bp, gn, bn were higher and rp, rn were lower in UCC and UDC than those in UDC(P<0.05).Ⅱ. The discriminant results of bladder urothelial carcinoma exfoliated cells with function based on chromatic parametersThe function was established on account of chromatic parameters of Rp, Gp, Bp, Rn, Gn, Bn, rp, rn, gp, bp, gn and bn of 1077 (training group) exfoliated cells in urine as follow:The Y1, Y2 and Y3 of function (1) discriminate uroepithelium normal exfoliated cells, urothelial carcinoma exfoliated cells and bladder urothelial carcinoma exfoliated cells respectively, the rate of back discriminant of function (1) was 71.2%. The discriminant rate of 248 exfoliated cells (identification group) in urine and 107 cases was 73.0% and 84.1%.Ⅲ. The test results of morphologic parameters for cytoplasm and nuclear of UCC, UDC and UNC in exfoliated cells of urine1.The UNC group has the highest values of RFFc, Spn, FIIc, the UDC group takes the second place, and the UCC group has lowest values; The UCC group has the highest values of Rnp, dmaj;n,RFFc,EEn, the UDC group takes the second place, and the UNC group has lowest values; The UNC group has the highest values of Ac,Ap,dmaj,c, dmin,c, Dc, Cc, ARn, Spc, EEc, the UCC group takes the second place, and the UDC group has lowest values; The UCC group has the highest values of An,dmin,n,Dn, Cn, ARc, the UNC group takes the second place, and the UDC group has lowest values; The UDC group has the highest values of PEn, the UNC group takes the second place, and the UCC group has lowest values; The UDC group has the highest values of PEc, FIIn, the UCC group takes the second place, and the UNC group has lowest values. The multiple comparison showed there were statistical significance in values of Rnp, dmaj,c, dmin.c, dmaj,n, dmin,n-Dc,Dn, Cc, Cn, PEc between UCC and UDC, there were statistical significance in values of Ac, Ap, An, Rnp, dmaj,c, dmin,c, dmaj,n, dmin,n, Dc, Dn, Cc, Cn, ARn, RFFc, RFFn, FIIn between UCC and UNC, and there were statistical significance in values of Ac, Ap, An, Rnp, dmaj,n, Dc, Dn, Cc, PEc, ARn, RFFc, RFFn, FIIn between UDC and UNC (P<0.05).2.The F value of Ac, Ap, An, Rnp, dmaj,c, dmin,c, dmaj,n, dmin,n,Dc,Dn, Cc, Cn, PEc, ARn, RFFc, RFFn, FIIn is respectively 480.608,536.658,113.755,747.099,541.269, 545.623,68.094,90.188,603.814,110.300,611.610,115.386,3.261,5.261,6.034, 10.763,3.648,the difference has statistic significance(P<0.05). The F value of PEn, ARc, Spc, Spn, EEc, EEn, FIIc is respectively 2.842,1.240,1.439,1.229,2.224,2.640, 0.410,the difference has not.statistic significance (P>0.05).IV. The discriminant results of bladder urothelial carcinoma exfoliated cells with function based on morphologic parametersThe functions were established on account of morphologic parameters of Ac, Ap, An,Rnp,dmaj.c,dmin,cdmaj,n, dmin.n,Dc,Dn,PEc,PEn,ARn,ARc,RFFc,RFFn, Spc, Spn, EEc, EEn, FIIn, FIIn 1077 (training group) exfoliated cells in urine as follow: The Y4, Y5 and Y6 of function (2) discriminate uroepithelium normal exfoliated cells, urothelial carcinoma exfoliated cells and bladder urothelial carcinoma exfoliated cells respectively, the rate of back discriminant of function (2) was 71.9%. The discriminant rate of 248 exfoliated cells (identification group) in urine and 107 cases was 70.2% and 81.3%.V. The discriminant results of bladder urothelial carcinoma exfoliated cells with function based on chromatic and morphologic parametersThe functions were established on account of chromatic parameters of Rp, Gp, Bp, Rn, Gn, Bn, rp, rn, gp, bp, gn, bn and morphologic parameters of Ac, Ap, An, Rnp, dmaj.c,dmin,c, dmaj,n, dmin,n,Dc, D n, Cc, Cn,PEc,PEn, ARn, ARc, RFFc, RFFn, Spc, Spn, EEc, EEn, FIIn, FIIn 1077 (training group) exfoliated cells in urine as follow: The Y7, Yg and Y9 of function (3) discriminate uroepithelium normal exfoliated cells, urothelial carcinoma exfoliated cells and bladder urothelial carcinoma exfoliated cells respectively, the rate of back discriminant of function (3) was 80.8%. The discriminant rate of 248 exfoliated cells (identification group) in urine and of 107 cases was 80.2% and 92.5%.Conclusions:1. Chromatic and morphologic features of exfoliated cells in urine stained by Pap test differ among UNC, UDC and UCC group. The difference reflect on chromatic parameters of Rp, Gp, Bp, Rn, Gn, Bn, rp, rn, gp, bp, gn, bn and morphologic parameters of Ac, Ap, An, Rnp, dmaj,c, dmin,c, dmaj,n, dmin,n, Dc, Dn, Cc, Cn, PEc, PEn, ARn, ARc, RFFc, RFFn, Spc, Spn, EEc, EEn, FIIn, FIIn. These parameters and their variance could be used to discriminate bladder urothelial carcinoma exfoliated cells.2. The chromatic parameters Rp, Gp, Bp, Rn, Gn, Bn, rp, gp and gn could be applied to discriminate the UNC, UDC and UCC group by Pap stain; the morphologic parameters Ap, An, Rnp, dmaj,c, dmin,c, dmaj,n,dmin,n, Cc,PEn and EEc could be applied to discriminate the UNC, UDC and UCC group; The chromatic parameters Rp, Rn, Bn, rp, rn, bn and the morphologic parameters Ap, An, Rnp, dmin,n, Dc, Dn, Cc, Cn, FIIc could be applied to discriminate the UNC, UDC and UCC group.3.Establishing the discriminant functions based on chromatic and morphologic parameters. The discriminant function (3) discriminate the UNC, UDC and UCC group, their coincidence rate of back discriminant,248 cells (identification group) and 107 cases were respectively 80.8%,80.2%,92.5%. The chi-square test showed the discriminant function (3) which based on chromatic and morphologic parameters is better than that based on chromatic or morphologic parameters (P＜0.05).