| Objective: This paper was aimed to quantitatively reveal the chromatic andmorphologic characters of the exfoliate cells of lung cancer in sputum smears stainedby Pap test, to find highly specific parameters applied to differentiate and classify theexfoliate cells, and to discuss the value of the chromatic and morphologic parameterson classify and discriminate the lung cancer cells.Methods: 160 cases sputum samples were collected from the patients diagnosed withlung cancer and healthy peoples. The exfoliate cells in sputum smears were dividedinto normal and lung cancer groups. The normal groups include the cells of columnarepithelium (CE), squamous epithelium (SqE), histiocytes (HC). Each category has 20cases and 120 cells. The lung cancer cells were divided into squamous cell carcinoma(SqCa), adenocarcinoma (AD), small cell carcinoma (SCC) and large cell carcinoma(LCC) according to WHO classification. They consisted of 204 SqCa cells, 156 ADcells, 147 SCC cells, and 85 LCC cells. The sputum samples were smeared usingconventional method, fixed by 95ï¼…alcohol, stained by Pap test. The images of 7category cells were taken a photograph in objective lense 40X of light microscopeequipped digital camera and inputted into the computer. Technique of computerimage analysis was applied to test their relative parameters. The chromatic parametersincluded three primary colors of cell (Rc, Gc, Bc) and their coefficient (rc, gc, bc), three primary colors of nuclear (Rn, Gn, Gn) and their coefficient (rn, gn, bn). Themorphologic parameters of cells and nuclear included area (Ac, An), circumference(Cc, Cn), Long axis (dmaj,c, dmaj,n), Short axis (dmin,c, dmin,n), Axis Ratio (Rac, Ran),Area of cytoplasm (Acp), Karyoplasmic ratio (Rnp), Form factor PE (PEc, PEn), Formfactor AR (ARc, ARn), Regular form factor (RFFc, RFFn), and Form irregular index(FIIc, FIIn). The values of the parameters were tested with one-way ANOVA andcompared with Least—significant Difference. The following groups were submittedto do the stepwise discriminate analysis. They were the normal and lung cancergroups, four subtypes of lung cancer cells groups, and three categories of normal cellsgroups. And then stepwise discriminate functions were established, the effect ofdiscriminate was evaluated.Results:â… . The test results of chromatic parameters for cell and it's nuclear in sputumsmears stained by Pap test1. The values of Rc, Rn of SqE are biggest, CE, LCC, HC, SCC, SqCa decreaseprogressively in turn, that of AD is smallest; The value of Gc of SqE is biggest, HC,SqCa, CE, AD, LCC decrease progressively in turn, that of SCC is smallest; Thevalues of Gn of SqE is biggest, HC, SqCa, SCC, AD, LCC decrease progressively inturn, that of CE is smallest; The value of Bc of SqE is biggest, HC, SCC, SqCa, CE,AD decrease progressively in turn, that of LCC is smallest; The value of Bn of SqE isbiggest, LCC, HC, AD, SqCa, SCC decrease progressively in turn, that of CE issmallest; The values of rc, rn of LCC are biggest, CE, SqE, HC, SCC, SqCa decreaseprogressively in turn, that of AD is smallest; The values of gc, gn of HE are biggest,SqCa, SqE, AD, SCC, CE decrease progressively in turn, that of LCC is smallest; Thevalue of bc of AD is biggest, SCC, SqCa, HC, CE, LCC decrease progressively in turn,SqE is smallest; The value of bn of AD is biggest, SCC, SqCa, CE, HC, LCC decrease progressively in turn, that of SqE is smallest. The F value of these parameters in 7kind of intercellular comparisons respectively are 69.00, 32.23, 64.52, 22.98, 32.94,7.35, 70.57,54.94, 48.63, 53.24, 29.31, 21.57 after ANOVA, the P value is 0.000, thedifference has statistics significance.2. The value of Rc, Rn, Gc, Gn, Bc, rc, rn of Normal cell are higher than the lungcancer cell, that of bc, bn is lower than the lung cancer cell, the F value is respectively169.18, 49.59, 123.30, 28.55, 72.72, 62.75, 33.11, 67.13, 49.48, the P value is 0.000,the difference has statistics significance. The gn in between the normal cell and thecancer cell the difference, P value is 0.0723. In normal cell group, the values of Rc, Rn, Gc, Gn, Bc and Bn of SqE are highest,the values of Gc, Rn, Gn, Bc and Bn of CE are smallest, the value of Rc of HC issmallest, the F value is respectively 42.62, 11.90, 292.43, 52.36, 127.47, 27.23, the Pvalue is 0.000, the difference has statistics significance. The value of Rc, rn of CE arehighest, SqE is next, HC is smallest; the values of Gc, gn of HC are highest, SqE isnext, CE is smallest; The value of Bc, bn are CE highest, HC is next, SqE is smallest.The F value respectively is 46.428, 11.04, 57.34, 217.72, 24.87, 9.23, the P value is0.000, and the difference has statistics significance.4. In cancer cell, the values of Rc, Rn, Gc, Gn, rc, rn of LCC are highest, thosevalues of bc, bn are lowest, the F value respectively is 36.566, 37.604, 16.214,6.563,80.657, 87.375, 24.710, 18.907, the P value is 0.000, the difference has statisticssignificance. The P value of Bn is 0.051.â…¡. The results of stepwise analysis on the exfoliate cells of lung cancer in sputumsmears on account of chromatic characters.The functions were established on account of chromatic parameters of Rc, Rn, Gc,Gn, Bc, Bn, rc, rn, gc, gn, bc, and bn as follow: The function of Y1-1,Y1-2 discriminate the normal and lung cancer cellsrespectively, their accurate rating were respectively 62.5ï¼…, 95.9ï¼…; the function ofY1-3, Y1-4, Y1-5, Y1-6 discriminate the cells of SqCa, AD, SCC and LCC respectively,their accurate rating were respectively 72.5ï¼…, 46.2ï¼…, 81.2ï¼…, 56.5ï¼…; The functionY1-7, Y1-8, Y1-9 discriminate the cells of CE, HC and SqE respectively, their accuraterating were respectively 100ï¼…, 95.8ï¼…, 94.2ï¼….â…¢. The results of stepwise analysis on the exfoliate cells of lung cancer in sputumsmears on account of morphologic characters.The functions were established on account of morphometry parameters of Ac,dmaj,c,dmin, c,Cc,An,dmaj,n,dmin, n,Cn,Acp, Rac, Ran,Rnp, PEc, ARc,RFFc, FIIc, PEn, ARn,RFFnå’ŒFIInas follow: The function of Y2-1, Y2-2 discriminate the normal and lung cancer cellsrespectively, their accurate rating were respectively 87.8ï¼…, 98.3ï¼…; the function of Y2-3,Y2-4, Y2-5, Y2-6 discriminate the cells of SqCa, AD, SCC and LCC respectively, theiraccurate rating were respectively 53.8ï¼…, 63.4ï¼…, 52.7ï¼…, 100.0ï¼…; The function Y2-7,Y2-8, Y2-9 discriminate the cells of CE, HC and SqE respectively, their accurate ratingwere respectively 100.0ï¼…, 100ï¼…, 97.3ï¼….â…£. The results of stepwise analysis on the exfoliate cells of lung cancer in sputumsmears on account of chromatic and morphologic characters.The functions were established on account of chromatic parameters of Rc, Rn, Gc,Gn, Bc, Bn, rc, rn, gc, gn, bc, bn and morphometry parameters of Ac,d(maj,c),dmin, c,Cc,An,dmaj,n,dmin, n,Cn,Acp,Rac,Ran,Rnp,PEc,ARc,RFFc,FIIc,PEn,ARn,RFFnå’ŒFIInas follow: The function of Y3-1, Y3-2 discriminate the normal and lung cancer cells respectively,their accurate rating were respectively 87.1ï¼…, 98.5ï¼…; the function of Y3-3, Y3-4, Y3-5, Y3-6 discriminate the cells of SqCa, AD, SCC and LCC respectively, their accuraterating were respectively 69.7ï¼…, 75.2ï¼…, 85.5ï¼…, 100.0ï¼…; The function Y3-7, Y3-8, Y3-9discriminate the cells of CE, HC and SqE respectively, their accurate rating were all100.0ï¼….The values of Rc, Gc, Bc, rc, Rn, Gn, and rn of the normal groups are higher thanthose of the lung cancer groups, but the values of bc and bn are lower than those of thelung cancer groups. The values of Rc, Gc, Rn, Gn, Bc and Bn of SqE are the highest innormal groups, but the values of Gc, Rn, Gn, Bc and Bn of CE are the lowest. Thevalues of Rc, Gc, Bc, Rn, Gn, rc and rn of LCC are the highest in cancer groups, but thevalues of bc and bn of LCC are the lowest. Statistic test show significance amongdifferent ceil groups.Conclusions:1. Chromatic and morphologic features of exfoliate cell in sputum smearsstained by Pap test differ not only between the normal and the lung cancer groups, butamong the normal groups, and among the lung cancer groups. The difference reflecton the parameters Rc, Rn, Gc, Gn, Bc, Bn, rc, rn, gc, gn, bc, bn and Ac, dmaj, c, dmin,c, Cc,An, dmaj, n, dmin, n, Cn, Acp, Rac, Ran, Rnp, PEc, ARc, RFFc, FIIc, PEn, mRn, RFFn andFIIn. These parameters and their variance could be used to classify and discriminatethe exfoliate cells.2. The parameters Ac, dmaj, c, dmin, c, Cc, An, dmaj, n, dmin, n, ARc, PEc, RFFc, FIIc,PEn, ARn and Rn could be applied to discriminate the normal and lung cancer cells;the parameters Ac, dmaj, c, dmin, c, Cn, Acp, Rnp, Rac, PEn, ARn, RFFn, FIIn, rn, gc, bc andbn could be applied to discriminate the cells of SqCa, AD, SCC and LCC; theparameters dmin, c, Cn, Rac, ARc, Gc, Gn, Bc and bn could be applied to discriminate thecells of CE, HC and SqE.3. Establishing the stepwise discrimination functions based on chromatic and morphologic parameters. The function of Y3-1, Y3-2 discriminate the normal and lungcancer cells respectively, their accurate rating were respectively 87.1ï¼…, 98.5ï¼…; thefunction of Y3-3, Y3-4, Y3-5, Y3-6 discriminate the cells of SqCa, AD, SCC and LCCrespectively, their accurate rating were respectively 69.7ï¼…, 75.2ï¼…, 85.5ï¼…, 100.0ï¼…;The function Y3-7, Y3-8, Y3-9 discriminate the cells of CE, HC and SqE respectively,their accurate rating were all 100.0ï¼…. Their effects must surpass the stepwisediscrimination function which establishes based on the chromatic or the geometrymorphologic parameters.4. A strategy was established that discriminate the exfoliate cells in sputumsmears according to chromatic and morphologic feature. Firstly, to discriminate thenormal and lung cancer cells applying the function (3-1); Secondly, to discriminatethe cells of SqCa, AD, LCC, and SCC; Finally, to discriminate the cells of CE, HC,and SqE. |
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