Epstein-Barr virus (EBV), a common pathogen in humans, is suspected as the cause of multiple pregnancy-related pathologies including depression, preeclampsia, and stillbirth. Moreover, transmission of EBV through the placenta has been reported. However, the focus of EBV infection within the placenta has remained unknown to date. In this study, we proved the expression of latent EBV genes in the endometrial glandular epithelial cells of the placenta and investigated the cytological characteristics of these cells. Sixty-eight placentas were obtained from pregnant women. Tissue microarray was constructed. EBV latent genes including EBV-encoding RNA-1 (EBER1), Epstein-Barr virus nuclear antigen 1 (EBNA1), late membrane antigen (LMP1), and RPMS1 were detected with silver in situ hybridization and/or mRNA in situ hybridization. Nuclear features of EBV-positive cells in EBV-infected placenta were compared with those of EBV-negative cells via image analysis. Sixteen placentas (23.5%) showed positive expression of all 4 EBV latent genes; only the glandular epithelial cells of the decidua showed EBV gene expression. EBV infection status was not significantly correlated with maternal, fetal, or placental factors. The nuclei of EBV-positive cells were significantly larger, longer, and round-shaped than those of EBV-negative cells regardless of EBV-infection status of the placenta. For the first time, evidence of EBV gene expression has been shown in placental tissues. Furthermore, we have characterized its cytological features, allowing screening of EBV infection through microscopic examination.
Decidua ; Depression ; Epithelial Cells ; Epstein-Barr Virus Infections ; Female ; Gene Expression ; Herpesvirus 4, Human* ; Humans* ; Image Cytometry ; In Situ Hybridization ; Mass Screening ; Membranes ; Pathology ; Placenta* ; Pre-Eclampsia ; Pregnant Women ; RNA, Messenger ; Silver ; Stillbirth ; Virus Latency

DNA, Neoplasm ; Humans ; Image Cytometry ; Pleural Effusion, Malignant ; diagnosis

Estimation of postmortem interval (PMI) is one of the difficult problems in forensic medicine. With the development of molecular biological techniques, DNA quantification methods were widely applied in estimating PMI. The postmortem degradation of DNA in different tissues and organs was discussed in this article and the recent DNA quantitative techniques being used for estimating PMI were reviewed. These techniques included single cell gel electrophoresis, Feulgen staining image analysis, flow cytometry.
Animals ; Cell Nucleus/metabolism* ; Comet Assay/methods* ; DNA/analysis* ; DNA Degradation, Necrotic ; Flow Cytometry ; Forensic Pathology ; Humans ; Image Processing, Computer-Assisted ; Kidney/pathology* ; Postmortem Changes ; Rosaniline Dyes ; Spleen/pathology* ; Temperature ; Time Factors

OBJECTIVETo discuss the application of automated DNA image cytometry (ICM) and fluorescence in situ hybridization (FISH) in the diagnosis of urothelial carcinoma of bladder.

METHODSFrom August 2008 to March 2009, 60 volunteers with informed consent were divided into two groups, 40 patients proven as urothelial carcinoma of bladder by pathology and 20 healthy individuals as control. Urine was collected and tested by cytology, ICM and FISH.

RESULTSOverall sensitivity of FISH was significantly higher in detection of malignancy than that of ICM (82.5% vs 62.5%, P < 0.05) and that of urine cytology (82.5% vs 25.0%, P < 0.05), while ICM was more sensitive to diagnose urothelial carcinoma of bladder than urine cytology (62.5% vs 25.0%, P < 0.05). Specificities of urine cytology, ICM and FISH were 100% in diagnosis of urothelial carcinoma of bladder (P > 0.05). Sensitivities of urine cytology, ICM and FISH have no correlation with pathological stage (P > 0.05), but have significant correlation with grade (P < 0.05).

CONCLUSIONSICM and FISH have the same specificity as urine cytology in diagnosis of urothelial carcinoma of bladder, but they have significantly higher sensitivity than urine cytology. FISH has the highest sensitivity among three diagnostic methods. Therefore, FISH may become a newly non-invasive technique for the diagnosis and surveillance of urothelial carcinoma of bladder.

Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Image Cytometry ; methods ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Sensitivity and Specificity ; Urinary Bladder ; pathology ; Urinary Bladder Neoplasms ; diagnosis ; pathology

OBJECTIVETo evaluate efficacy of sputum imaging cytometry in early diagnosis on lung cancer among tin miners exposed to dust and analyze possible risk factors related to lung cancer among tin miners.

METHODSFrom a cohort of tin miners in Guangxi Province, a total of 345 male tin miners higher than 45 years old and with high exposure to crystalline silica dust were randomly selected as the objects. Imaging cytometry was used to implement the sputum analysis for the screening on lung cancer according to the experience diagnostic standard. All objects were then followed up to the end of 2006. Clinical diagnosis of lung cancer was used as the golden standard to evaluate the efficacy of screening.

RESULTSFrom 1998 to 2006, 11 new cases were diagnosed as clinical lung cancer. Except of age and exposure to occupational hazards, smoking status (P = 0.0384) and mean smoking dose (P = 0.0078) were significantly associated with lung cancer, and the adjusted odds ratio of high level to the low was 18.21 (2.15 approximately 154.39). The sensitivity, specificity and Youden's index of the sputum imaging cytometry for the experience diagnosis were 27.3%, 83.9% and 11.2% respectively. According to the ROC curve analysis, area under ROC Curve (AUC) of C2.5 (the percentage when the DNA index ranged from 1.25 to 2.50) was 0.647 (0.525 approximately 0.768), with the optimal operating point (OOP) of 1.70%. Sensitivity, specificity, agreement rate, positive predictive value, negative predictive value and Youden's index for predicting lung cancers in high-exposure tin miners were found to be 72.7%, 62.3%, 62.6%, 6.0%, 98.6% and 35.0% respectively.

CONCLUSIONSmoking is confirmed as an important risk factor of lung cancer in tin miners. The diagnostic efficiency can be improved if the diagnostic point of C2.5 is adjusted to 1.70%.

Aged ; China ; Cohort Studies ; Dust ; Humans ; Image Cytometry ; Lung Neoplasms ; diagnosis ; etiology ; pathology ; Male ; Mass Screening ; Middle Aged ; Mining ; Occupational Exposure ; adverse effects ; Risk Factors ; Sampling Studies ; Smoking ; adverse effects ; Sputum ; cytology ; Tin

Determination of postmortem interval (PMI) is one of the most valuable subjects in forensic practice. It, however, is often very difficult to accurately determine the PMI in daily practice. Forensic DNA technology has recently been used to estimate the PMI. It has certain advantage to traditional methods. This article reviews this technology with respect to its invention, development, advantage, disadvantage, and potential future applications with emphasis on correlation of DNA degradation and PMI.
Animals ; Bone Marrow Cells/metabolism* ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Flow Cytometry ; Forensic Medicine/methods* ; Hepatocytes/metabolism* ; Humans ; Image Processing, Computer-Assisted/methods* ; Myocardium/metabolism* ; Postmortem Changes ; Spleen/metabolism* ; Time Factors

OBJECTIVETo compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions.

METHODS87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry.

RESULTSOf the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method.

CONCLUSIONThe study demonstrated that DNA-ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions.

Aneuploidy ; Cell Nucleus ; metabolism ; Cervical Intraepithelial Neoplasia ; diagnosis ; genetics ; metabolism ; DNA, Neoplasm ; analysis ; Female ; Humans ; Image Cytometry ; methods ; Immunohistochemistry ; Ki-67 Antigen ; analysis ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; diagnosis ; genetics ; metabolism

PURPOSE: To investigate the relationship between optical coherence tomography (OCT) and scanning laser polarimetry (SLP) in measuring peripapillary retinal nerve fiber layer (RNFL) thickness in glaucomatous eyes. METHODS: Fifty glaucomatous eyes were evaluated in this study. Evaluations were analyzed two ways. First, parameters of the Stratus OCT (average thickness, superior/ inferior average) and GDx VCC (TSNIT average, nerve fiber indicator (NFI), superior/ inferior average) were correlated using the Pearson's correlation coefficient (r). Secondly, comparison (r) of these parameters was completed using the mean deviation (MD) of visual field defect. RESULTS: The following parameters were found to be significantly correlated (P<0.005). TSNIT average/average thickness (r=0.673), NFI/average thickness (r=-0.742), superior average (r=0.841), and inferior average (r=0.736). In the correlation analysis using the severity of visual field defect, all these parameters had statistically meaningful correlations (P<0.005). CONCLUSIONS: GDx VCC and Stratus OCT are highly correlated in glaucomatous eyes. Therefore, peripapillary RNFL thickness measured by Stratus OCT and GDx VCC may be equally helpful in the diagnosis of glaucoma.
Tomography, Optical Coherence/*methods ; Severity of Illness Index ; Retina/*pathology ; Reproducibility of Results ; Middle Aged ; Male ; Laser Scanning Cytometry/*methods ; Humans ; Glaucoma/*pathology ; Female ; Aged, 80 and over ; Aged ; Adult

Aneuploidy ; Animals ; Cervical Intraepithelial Neoplasia ; genetics ; pathology ; DNA, Neoplasm ; analysis ; Diagnosis, Differential ; Female ; Image Cytometry ; methods ; Neoplasms, Squamous Cell ; genetics ; pathology ; Prognosis ; Uterine Cervical Neoplasms ; genetics ; pathology

OBJECTIVE: To investigate the relationship between the DNA degradation in cells and postmortem interval. METHODS: Tissue pieces of the heart, liver, spleen and kidney of one corpse (male, 54 years old, died of mechanical injury, PMI of 6h) were obtained in 6, 12, 24, 48 h after death, fixed in Carnoy fluid, and then paraffin sections were prepared, stained with Feulgen and analyzed by Image analysis technology (IAT). Meanwhile the single-cell suspension of tissues of the man was prepared and inspected by FCM after PI stained. RESULTS: The amount of DNA of in heart, liver and kidney of human had a repid degraded in first 6 hours after death, that in the spleen showed a better relationship between DNA degradation and PMI. The results was verified by FCM and IAT. CONCLUSION The degradation of DNA of and human tissues shows a well relationship with PMI, especially in spleen. It is useful in estimation of PMI.
Cell Nucleus/metabolism* ; DNA/metabolism* ; Flow Cytometry ; Forensic Medicine/methods* ; Humans ; Image Processing, Computer-Assisted/methods* ; Kidney/metabolism* ; Liver/metabolism* ; Male ; Middle Aged ; Myocardium/metabolism* ; Postmortem Changes ; Spleen/metabolism* ; Staining and Labeling/methods* ; Time Factors