Explore Of Analyzing DNA Ploidy Of The Cervical Cast-off Cell In Cervical Neoplasia Screening

Purposes:Part one:To observe the effect of automatic and manual slicing methods and tissue types on the thickness variation of paraffin-embedded sections to provide a reference for observing the organization or the cell's morphology and the position or composition of chemical substances and also for the evaluation of the quantitative analysis results.Part two:(1) Application of cytology,pathology diagnosis,nuclear DNA ploidy analysis and second-generation hybrid capture test used to detect HPV-DNA,and make comparative analysis.To provide an effective and volant adjuvant diagnostic method for cervical cancer and cervical dysplasia screening.(2) To provide an effective screening method for atypical squamous cells of undetermined significance cases which have the potential risk of malignant transformation,and to provide much more evaluating information for prognosis and evaluation of therapeutic efficacy.Methods:Part one:Liver,pancreas,stomach and lung samples were obtained from five healthy adult male rats.Each tissue was separately embedded in paraffin.Slices were cut automatically and manually at various thickness set points,3μm,5μm,7μm,10μm,by an automatic microtome.After the re-embedding vertical slicing and HE staining,the thickness of these slices was measured by a TIGER image analysis system.Part two:Each of cervical samples from the women who accepted the conventional cast cell screening was made into liquid-based smear sample.The smear were separated into two groups for the experiment.One group were stained by papanicoloau stain to make conventional cytology screening.The other group were stained by Feulgen method and was analyzed by nuclear DNA ploidy analysis system.Chose the samples which were regarded as doubtful cases diagnosed by cytologic analysis, these samples were delivered to the pathohistology.The results of samples which were infected with human papillomavirus(HPV) were compared with those of DNA ploidy analysis in combination with the level of cervical lesions and age factor.Results:Part one:(1) Only the automatic or manual slicing procedure exerted effect on the slice thickness.(2) The mean measured thickness of each tissue section was bigger than the microtome set point.On condition of the same set point,the mean measured thickness of automatic section was smaller than manual section.(3) No matter automatic or manual slice,the measured thickness has a direct ratio tendency to bigger along with the microtome set point become bigger.(4) The thickness of auto-slice and manual slice were 29%～50%and 48%～91%, respectively,thicker than the nominal thickness set on the microtome.There is a 25%difference between the thicknesses of automatically cut slices and manually cut slices.The relative error rate of the two slicing methods is 61%.(5) Dispersion tendency of the measured thickness of the same kind of tissue had an escalating trend along with the microtome set point become bigger,the rang was bigger when cut manual than cut automatic.Part two:(1) 362 abnormal cases from 370 doubtful cases were suspected ones,including 140 cases of atypical squamous intraepithelial undermined significance(ASCUS),190 cases of low-grade squamous intraepithelial lesion(L-SIL) and 31 eases of highgrade squamous intraepithelial lesion(H-SIL),were diagnosised by conventional cytology examination.23 eases of carcinoma in situ(CINⅢ) and 49 CINⅡwere ??detected among 370 women by pathohistology examination.258 abnormal cases were diagnosised by DNA image analysis.According to the criterion of pathohistology diagnosis,it can say that the sensitivity of DNA ploidy analysis was 98.6%,and sensitivity of conventional cytology was 72.2%.(2) 191 cases in 370 patients made the HPV test,in which 81 cases were infected HR-HPV.Comparing the results of HPV detection with those of DNA ploidy analysis,there were a correlation between their positive detection rates and degree of dysplasia.Almost all cases with infection of high-risk human papillomavirus (HR-HPV) were aneuploidy.(3) HPV test was made in 191 cases,calculated by the pathology diagnosis as the gold-standard,the sensitivity of nuclear DNA ploidy analysis in screening for cervical lesions higher than CINⅡwas 84.4%,while the HR-HPV detection was 75.6%.(4) There was a directly correlation between the infection rate of HR-HPV and the age of patients.Youth group was higher than the older group.While the prevalence of aneuploidy was not influenced by the age of patients.(5) Cytology diagnosed as uncertain cases such as ASCUS,the patients who had infected HR-HPV and aneuploid were tested positive,the detection rate was highest in equal or higher than CINⅡgroup.On the reverse side,no cases was higher than CINⅠ.Conclusions:Part one:There is a 25%difference between the thicknesses of automatically cut slices and manually cut slices.The relative error rate of the two slicing methods is 61%.The precision of automatic cut thickness is higher,therefore,is better than manual sectioning.Part two:(1) Nuclear DNA ploidy analysis is better for screening precancerous and carcinoma of the uterine cervix than conventional cytology.Provide an effective and volant adjuvant diagnostic method for cervical cancer screening. (2) HR-HPV infection rate is increased as well as the HR-HPV positive detection rates of the youth group is higher than in middle-aged group,make important part in younger-cervical-cancer.(3) Aneuploidy test results have no difference between the patient's vary age,while the positive of HR-HPV make the sense.Most of infected cases can find the aneuploidy,while there are only part of aneuploidy cases have infected HR-HPV. The sensitivity of DNA ploidy analysis is higher than the HR-HPV test.(4) HR-HPV infection rate is increased as the cytology or pathology detection of cervical lesion become higher.(5) Make reference for the differentiate and diagnosis of the cytologic uncertain cases such as ASCUS.