The Role Of Nuclear DNA Ploidy Analysis Of Cervical Cast-off Cells In Cervical Intraepithelial Neoplasia Screening

Purposes:Part one:On the theoretical basis of the ratio of DNA content in volume unit of whole diploid, tetraploid and octoploid nuclei was 2 or 4, we re-estimate the reliability of measuring the DNA content in volume unit of whole hepatocyte nuclei on the liver tissue in situ by image analysis.Part two:(1) 487 cases of cervical smears were for DNA ploidy analysis by TIGER-cell image analysis system, and we compared the result of DNA polidy analysis to observe the stability and accuracy of the cell image analysis system.(2) The results of cytology, pathology diagnosis, nucleai DNA polidy analysis and cape medical nucleic acid molecules using rapid hybridization kits for detection of HPV genotyping had been compared to provide an effective and volant adjuvant diagnostic method for cervical cancer screening; and the factor caused the age of cervical cancer becoming younger.(3) With biopsy results as the standard, we compared and anlysised the resuls of four kinds of testings to provide an effective identifying method for clinical borderline cases and uncertain cases on pathological diagnosis, and to provide much more evaluating information for prognosis, treatment and evaluation of therapeutic efficacy.Methods:Part one:The 4μm and 11μm liver tissue slices were prepared respectively, from 10 healthy adult male Kunming mice, and then together stained with modified Feulgen stain. The DNA content in volume unit of whole diploid or tetraploid or octoploid nuclei were measured and calculated by TIGER-cell image analysis system respectively, with two methods.Part two:Each of cervical samples from the 487 women who accepted the convertional cytological screening was made into two smears by TCT. One sample stained by papanicolaous stain was diagnosed by conventional cytology. The other sample stained by Feulgen method was analyzed by muclear DNA polidy anlysis. The uncertain cases were required to be deteced by biopsy. The samples which had the doubt of infecting with human papillomavirus should be tested by HR-HPV testing. Then we compared the results of the conventional cytology, biopsy, HR-HPV testing and those of DNA plidy analysis, and did the research and analysis of the age groups, and so with the results of HR-HPV testing and DNA polidy analysis.Result:Part one:The ratio of DNA content in volume unit of whole diploid, tetraploid and octoploid nuclei were close to 2 or 4 in liver tissue of each Kunming mouse by the 3D testing, and the results had not statistically significant with the theoratic mupltiples.Part two:(1) 487 cases of the appointment of women were tested by conventional cervical cytology, 97.33% were abnormal, including 168 cases of ASCUS,257 cases of LSIL and 49 cases of HSIL, were diagnosed by conventional cytology examination.220 CINⅠ,72 CINⅡand 55 cases of carcinoma in situ were detected by pathohistology examination.337 abnormal cases were diagnosed by DNA image analysis. According to the criterion of pathohistology diagnosis, it could say that the sensitivity of DNA polidy analysis was 92.1%, and that of convertional cytology was 75.6%.(2) 210 cases were detected by HR-HPV testing, in which 97 cases were infected. The difference of positive rate of the youth group and middle-age group had statistical significance. The positive rate of the youth group was more than that of the middle-age group.(3) According to the criterion of pathohistology diagnosis, we compared the results of the HR-HPV testing, conventional cytology and DNA polidy anlysis of 210 cases. The differences of results of DNA ploidy analysis and HR-HPV testing at different levels of pathological diagnosis had statistically significant in overall. The sensitivity of the cervical shedding nuclear DNA polidy analysis was 86.0% and the specificity of it was 36.3%. The sensitivity of the HR-HPV testing was 75.0% and the specificity of it was 66.4%. The sensitivity of the conventional cytology was 81.3% and the specificity of it was 31.0%. (4) 140 ASCUS were detected more than twice. The pathohistology diagnosis was to be as the criterion.With the increase at the levels of pathology, the positivity of the DNA polidy analysis combined with the HR-HPV testing increased. The level of the CINⅡis the highest. With the results of HR-HPV and DNA polidy analysis was negative, the highest rate of the pathological diagnosis is≤CINⅠ, and≥CINⅡwas 0.Conclusions:Part one:The 3D testing can obtain the total DNA content in volume unit of whole hepatocyte nuclei on the tissue slices stably. With using an image analysis system, we can obtain the content of chemical substances in volume unit of whole cells or nuclei on the tissue in situ by suitable sampling and calculation.Part two:(1) DNA polidy analysis combine with the conventional cytology can improve the sensitivity of the cervical intraeopthelial neoplasia screening.(2) The increase of HR-HPV infection and the higher positive of youth group's HR-HPV infection were perhaps an important factor in making cervical cancer become younger.(3) HR-HPV testing combine with the DNA polidy analysis can improve the sensitivity and the specificity of the cervical intraeopthelial neoplasia screening.(4) Application of the biopsy, HR-HPV testing and DNA polidy analysis can make reference for the diagnosis and clinic treatments of the cytologic uncertain cases such as ASCUS, and we can obtain more information of the lesions than the morphology.