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Objective To investigate the clinical value of quantitative detection of DNA aneuploidy in the diagnosis and treatment of cervical lesions in middle-aged and senile women.Methods A total of 1 404 middle-aged and elderly women who underwent screening for early cervical lesions were retrospectively studied.Patients were divided into the two groups:the 40-49 years old group(n=897)and the 50-78 years old group(n=507).Cervical lesions were screened by DNA ploidy analysis and the results were compared with those screened by liquid-based cytology,colposcopy and high-risk human papillomavirus(HPV).Results The positive detection rate of HPV by DNA ploidy analysis was 54.4 ％ (764/1 404).Of 1 404 patients,HPV16/18 infection accounted for 21.3％ (299/1 404).The detection rate of heteroploid cells was 50.92 ％ (715/1 404).There was a significant positive correlation between HPV infection type and cervical epithelial cell ploidy changes(r=870,P=0.001).The detection rate of HPV by liquid-based cytology was 45.08 ％,which was lower than that by DNA aneuploidy(x2=9.594,P=0.002).The differences in the incidences of low-grade squamous intraepithelial lesion(LSIL)and high-grade squamous intraepithelial lesion(HSIL)and above categories of lesions were statistically significant (x 2 =289.598,P =0.000) between patients with and without DNA aneuploidy.The statistically significant differences were found between the 40-49 years old group and 50-78 years old group(P＜0.05)in the occurrence of abnormal DNA ploidy cells,HPV infection rate,the proportion of LSIL,HSIL and above categories of lesions.Conclusions Compared with the conventional cytology,DNA aneuploidy quantitative detection has higher sensitivity and better specificity,and has no significant difference from the high-risk HPV detection.It can be used as one of methods for screening cervical lesions in middle-aged and elderly women,especially those with highrisk HPV infection.
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Objective@#To investigate the clinical value of quantitative detection of DNA aneuploidy in the diagnosis and treatment of cervical lesions in middle-aged and senile women.@*Methods@#A total of 1 404 middle-aged and elderly women who underwent screening for early cervical lesions were retrospectively studied.Patients were divided into the two groups: the 40-49 years old group(n=897)and the 50-78 years old group(n=507). Cervical lesions were screened by DNA ploidy analysis and the results were compared with those screened by liquid-based cytology, colposcopy and high-risk human papillomavirus(HPV).@*Results@#The positive detection rate of HPV by DNA ploidy analysis was 54.4%(764/1 404). Of 1 404 patients, HPV16/18 infection accounted for 21.3%(299/1 404). The detection rate of heteroploid cells was 50.92%(715/1 404). There was a significant positive correlation between HPV infection type and cervical epithelial cell ploidy changes(r=870, P=0.001). The detection rate of HPV by liquid-based cytology was 45.08%, which was lower than that by DNA aneuploidy(χ2=9.594, P=0.002). The differences in the incidences of low-grade squamous intraepithelial lesion(LSIL)and high-grade squamous intraepithelial lesion(HSIL)and above categories of lesions were statistically significant(χ2=289.598, P=0.000)between patients with and without DNA aneuploidy.The statistically significant differences were found between the 40-49 years old group and 50-78 years old group(P<0.05)in the occurrence of abnormal DNA ploidy cells, HPV infection rate, the proportion of LSIL, HSIL and above categories of lesions.@*Conclusions@#Compared with the conventional cytology, DNA aneuploidy quantitative detection has higher sensitivity and better specificity, and has no significant difference from the high-risk HPV detection.It can be used as one of methods for screening cervical lesions in middle-aged and elderly women, especially those with high-risk HPV infection.
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Objective To explore the application value of multiplex ligation-dependent probe amplification (MLPA) for diagnosing chromosome aneuploid abnormality diseases.Methods Seven hundred and thirty cases of prenatal samples were tested with MLPA detection and by G-band karyotype analysis.If the findings from the samples were different by two methods,they were then detected by fluorescence in situ hybridization (FISH).The sensitivity,specificity and positive predictive value of MLPA were calculated.Results There were 709 normal euploidies,12 cases of Down syndrome,4 cases of Eswards syndrome,1 case of Patau syndrome,1 case of Turner syndrome,1 case of 47,XXX,1 case of 47,XYY,1 case of 46,XY/47,XY + 21.The sensitivity,specificity and positive predictive value of MLPA were respectively 95％,100％ and 100％,respectively.Conclusion MLPA is a rapid and efficient method for analyzing aneuploids and has the good value of clinical applications.
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Objective In order to investigate the ability of flow cytometer (FCM) to diagnose malignant tumor and evaluate prognosis of tumors in salivary glands, the DNA ploidy and cell cycle had been analysed and the association of these parameters with histologic grades, lymph node metastasis were studied. Methods The fresh tissues of benign tumor, mixed tumor and malignant tumor were detected by FCM. Results DNA aneuploid has not been detected in benign tumors; 44 cases of mixed tumor has been detected with DNA aneuploid nd with high SPF; it suggests that these cases have malignant trend; most of malignant tumor were detected with high aneuploid. DI was not correlated with lymph node metastases, but correlated with histologic grades (P
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OBJECTIVE: The aim of this study was to determine whether fetal nucleated red blood cells (NRBCs) could be distinguished from maternal cells in peripheral blood using an erythroblast scoring system. Presumptive fetal NRBCs were further analyzed through the use of fluorescent PCR amplification with polymorphic STR markers to prove fetal origin. METHODS: NRBCs were isolated by density gradient separation, CD15/45 depletion, and gamma hemoglobin positive selection from peripheral blood of seven women who had undergone termination of pregnancy because of fetal trisomy 21 (n=4), 18 (n=1), and 13 (n=2). Candidate fetal NRBCs, based on four discrete morphological and hemoglobin staining criteria, were then subjected to fluorescent PCR amplification of chromosome 21 short tandem repeat (STR) markers (D21S1411, D21S11) and chromosome 18 STR markers (D18S535). RESULTS: In all cases candidate fetal NRBCs were accurately identified based on erythroblast scoring system and confirmed to be fetal in origin based on the presence of shared and non-shared polymorphic DNA alleles when compared to DNA isolated from maternal cells. Also in five cases aneuploid fetal cells in maternal blood were identified through the use of fluorescent PCR amplification with polymorphic STR markers. CONCLUSION: We were able to distinguish fetal NRBCs from maternal cells and prove fetal origin independent of gender. These results suggest that this novel combined approach to fetal cell isolation through using an erythroblast scoring system and genetic analysis by STR analysis is a promising method for noninvasive prenatal diagnostic applications.
Subject(s)
Female , Humans , Pregnancy , Alleles , Aneuploidy , Cell Separation , Chromosomes, Human, Pair 18 , Chromosomes, Human, Pair 21 , DNA , Down Syndrome , Erythroblasts , Erythrocytes , Microsatellite Repeats , Polymerase Chain ReactionABSTRACT
Flowcytometry is a very important technique for the analysis of cell properties, with the advantages of simultaneous multiparameter analysis of large cell population in a short time. Recent advances in computer science and techniques in cell preparation and staining make it more valuable for the study of cell biology and its clinical application. This study was performed to establish the techniques of flowcytometry analysis of osteosarcoma cells, to evaluate the results of the characteristics of the DNA and specific cell cycle phase of osteosarcoma cells obtained by preparation of paraffin-embedded tissue blocks, and to analyze any possible difference between cell populations lacated apart from each other in the tumor mass for making a base for further clinical application. Paraffin-embedded tissue blocks were obtained from 10 cases of primary osteosarcoma, which had undergone amputation without chemotherapy or radiotherapy. Tissue blocks obtained from the most superficial and the deepest portions of the tumor mass from the skin surface were selected respectively in each cases. To evaluate the technique and results obtained, analysis of the whole sample were performed twice in a separate setting. Satisfactory DNA histogram was obtained from 14 of 20 tissue blocks, with the values of distribution in the specific cell cycle phases. DNA aneuploidy was found in 2 cases with a DNA index of 1.6 and 1.3, and no difference in DNA ploidy by the location in tumor mass. The S-phase and G2+M phase fraction were 13.2±8.5 and 6.2±3.1 respectively, reflecting the increased cell proliferation compared with normal cell population. There was no statistically significant difference of these values between superficial and deep portions, but the difference was 9.0±9.7 with a maximum of 26.6, much greater than the difference 3.3±3.6, between the first and second set of analysis. Flowcytometry is a very useful technique in the analysis of the DNA and cell cycle phase properties, and the characteristics of DNA and cell proliferation status of osteosarcoma cells were successfully evaluated by this technique. Unsatisfactory DNA histograms were thought to be the result of inappropriate samples. To adequately evaluate the changes in the tumor mass, standardization in obtaining tumor tissue about the location in the tumor mass is suggested for future studies with flowcytometry about the properties of tumor cells.
Subject(s)
Amputation, Surgical , Aneuploidy , Cell Cycle , Cell Proliferation , DNA , Drug Therapy , Osteosarcoma , Ploidies , Radiotherapy , SkinABSTRACT
A flow cytometric analysis of the nuclear DNA content of solid tumors using paraffin-embedded tissues has become available since 1983, and its ploidy pattern has been designated as an important prognostic parameter in many human tumors. Hepatocellular carcinoma(HCC) is one of the most common malignant tumors among Koreans, but little information is consolidated about the significance of ploidy pattern. We measured the nuclear DNA content of 62 surgically resected HCCs and 45 non-neoplastic tissues from the surrounding parenchyma by flow cytometry. Aneuploid was detected in 18 cases(29.0%) in HCCs and 2 cases(4.4%) in nonneoplastic hepatic parenchyma(p<0.005). Correlations between the DNA ploidy pattern and various clinicopathologic findings of HCCs were analized. The mean tumor size was significantly different(p<0.05) between the aneuploid group(8.8 cm) and the diploid group(6.1 cm). Mean age of the aneuploid group was younger(47 year) than the diploid group(51 years), but the difference was not statistically significant(p=0.052). The DNA pattern did not show any meaningful correlation with the gross and microscopic features of HCC except for the presence of capsule. These results suggest that DNA ploidy correlates with growth rate of the tumor and it may be a possibly useful prognostic factor in HCCs.