DNA image cytometry test for primary screening of esophageal cancer: a population-based multi-center study in high-risk areas in China

Objective： To evaluate the feasibility of DNA image cytometry （DNA-ICM） as a primary screening method for esophageal squamous cell cancer （ESCC）. Methods： A total of 5,382 local residents aged 40-69 years from three high-risk areas in China （Linzhou in Henan province, Feicheng in Shandong province and Cixian in Hebei province） from 2008 to 2011 were recruited in this population-based screening study. And 2,526 subjects declined to receive endoscopic biopsy examination with Lugol＇s iodine staining, while 9 and 815 subjects were excluded from liquid-based cytology and DNA-ICM test respectively due to slide quality. Finally, 2,856, 5,373 and 4,567 subjects were enrolled in the analysis for endoscopic biopsy examination, liquid-based cytology and DNA-ICM test, respectively. Sensitivity （SE）, specificity （SP）, negative predictive values （NPV） and positive predictive values （PPV） as well as their 95% confidence intervals （95% CI） for DNA-ICM, liquid-based cytology and the combination of the two methods were calculated. Receiver operating characteristic （ROC） curves were applied to determine the cutoff point of DNA-ICM for esophageal cancer. Results： DNA-ICM results were significantly correlative with esophageal cancer and precancer lesions （X2= 18.016, P〈0.001）. The cutoff points were 5,802, 5,803 and 8,002 based on dissimilar pathological types of low grade intraepithelial neoplasia （LGIN）, high grade intraepithelial neoplasia （HGIN）, and ESCC, respectively, and 5,803 was chosen in this study considering the SE and SP. The SE, SP, PPV, NPV of DNA-ICM test （cutoff point 5,803） combined with liquid-based cytology [threshold atypical squamous cells of undetermined significance （ASCUS）] were separately 72.1% （95% CI： 70.3%-73.9%）, 43.3% （95% CI. 41.3%-45.3%）, 22.8% （95% CI： 21.1%-24.5%） and 87.0% （95% CI： 85.7%-88.3%） for LGIN, 85.7% （95% CI： 84.3%-87.1%）, 41.3% （95% CI： 39.3%-43.3%）, 4.6% （95% CI： 3.8%-5.4%） and 98.9% （95% CI： 98.5%-99.3%） for HGIN, and 96.0% （95% CI： 95.2%-96.8%）, 40.8% （95% CI： 38.8%-42.8%）, 1.7% （95% CI： 1.2%-2.2%） and 99.9% （95% CI： 99.8%-100.0%） for ESCC. Conclusions： It is possible to use DNA-ICM test as a primary screening method before endoscopic screening for esophageal cancer.

Detection of DNA Aneuploidy in Exfoliated Airway Epithelia Cells of Sputum Specimens by the Automated Image Cytometry and Its Clinical Value in the Identification of Lung Cancer

To evaluate the value of detecton of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry for the identification of lung cancer, 100 patients were divided into patient group (50 patients with lung cancer) and control group (30 patients with tuberculosis and 20 healthy people). Sputum was obtained for the quantitative analysis of DNA content of exfoliated airway epithelial cells with the automated image cytometry, together with the examinations of brush cytology and conventional sputum cytology. Our results showed that DNA aneuploidy (DI>2.5 or 5c) was found in 20 out of 50 sputum samples of lung cancer, 1 out of 30 sputum samples from tuberculosis patients, and none of 20 sputum samples from healthy people. The positive rates of conventional sputum cytology and brush cytology were 16 % and 32 %, which was lower than that of DNA aneuploidy detection by the automated image cytometry (P<0.01,P>05). Our study showed that automated image cytometry, which uses DNA aneuploidy as a marker for tumor, can detect the malignant cells in sputum samples of lung cancer and it is a sensitive and specific method serving as a complement for the diagnosis of lung cancer.

Development and validation of a risk model for noninvasive detection of cancer in oral potentially malignant disorders using DNA image cytometry

Objective:To elucidate whether DNA aneuploidy was an independent discriminator for carcinoma within oral potentially malignant disorders(OPMDs),and further establish and validate a risk model based on DNA aneuploidy for the detection of oral cancer.Methods:A total of 810 consecutive patients with OPMD were prospectively enrolled from March 2013 to December 2018,and divided into a training set(n=608)and a test set(n=202).Brushing and biopsy samples from each patient were processed by DNADNA image cytometry and histopathological examination,respectively.Results:DNA aneuploidy of an outside DNA index≥3.5 in OPMD was an independent marker strongly associated with malignant risk[adjusted odds ratio:13.04;95%confidence interval(CI):5.46-31.14].In the training and test sets,the area under the curve(AUC)was 0.87(95%CI:0.82-0.91)and 0.77(95%CI:0.57-0.97),respectively,for detecting carcinoma in OPMD patients.The independent risk factors of lateral/ventral tongue and non-homogenous type combined with a risk model built with a multivariate logistic regression revealed a more favorable diagnostic efficacy associated with the training set(AUC:0.93;95%CI:0.91-0.96)and test set(AUC:0.94;95%CI:0.90-0.98).The sensitivity and specificity of carcinoma detection within OPMD was improved to 100%and 88.1%,respectively.Conclusions:This large-scale diagnostic study established a risk model based on DNA aneuploidy that consisted of a noninvasive strategy with lateral/ventral tongue and non-homogenous features.The results showed favorable diagnostic efficacy for detecting carcinoma within OPMD,irrespective of the clinical and pathological diagnoses of OPMD.Multicenter validation and longitudinal studies are warranted to evaluate community practices and clinical applications.

Demands and technical developments of clinical flow cytometry with emphasis in quantitative,spectral,and imaging capabilities

As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow cytometry in laboratory hematology(e.g.,diagnoses of minimal residual disease and various types of leukemia),summarizes state-of-the-art clinical flow cytometers(e.g.,FACSLyricTMby Becton Dickinson,DxFLEX by Beckman Coulter),then considers innovative technical improvements in flow cytometry(including quantitative,spectral,and imaging approaches)to address the limitations of clinical flow cytometry in hematology diagnosis.Finally,driven by these clinical demands,future developments in clinical flow cytometry are suggested.

Morphological Effects of Natural Products on Schizosaccharomyces pombe Measured by Imaging Flow Cytometry

Gaining a full understanding of the mechanisms of action of natural products as therapeutic agents includes observing the effects of natural products on cellular morphology,because abnormal cellular morphology is an important aspect of cellular transformations that occur as part of disease states.In this study a set of natural products was examined in search of small molecules that influence the cylindrical morphology of fission yeast Schizosaccharomyces pombe.Imaging flow cytometry of large populations of S.pombe exposed to natural products captured cell images and revealed changes in mean length and aspect ratio of cells.Several natural products were found to alter S.pombe’s morphology relative to control,in terms of elongating cells,shrinking them,or making them more round.These results may facilitate future investigations into methods by which cells establish and maintain specific shapes.