| Objective:1.DNA-image cytometry and routine exfoliative cytological were used to detect pleural effusion.The results of the two methods were compared to evaluate the application value of DNA-image cytometry analysis in the diagnosis and identification of pleural effusion.2.By using DNA-image cytometry analysis to detect pleural effusion in patients with lung cancer,analyze the difference of positive detection rate in different subtypes of lung cancer,so as to explore the value of DNA-image cytometry analysis in diagnosing lung cancer.3.In this study,the exfoliated cells from the pleural effusion of the lung cancer patients are made into a cell block,combined with immunocytochemistry,application of cell block immunocytochemistry in the diagnosis and differentiation of lung cancer.In order to provide a new source of specimens for the subsequent EGFR and ALK gene detection of lung cancer patients who cannot obtain tissue samples.Methods: 1.The pleural effusion of 160 hospitalized patients received by the pathology department of the Qinghai Affiliated Hospital from December 2015 to January 2018 was collected.2.The number of DNA aneuploidy in pleural effusion was detected by DNA-image cytometry analysis,and the results were compared with human identification detection under microscope.3.55 cases of pleural effusions with residual pleural effusion which was diagnosised by conventional cytology as "visible cancer cells" and "suspected cancer cells" are made into cell blocks from the exfoliated cells in the pleural effusion.4.The cell blocks were sliced and stained with HE.The expression of CK,TTF-1,Napsin A,Cg A,Syn,P40,P63 and CR were detected by immunocytochemical assay to identify histological typing.5.Using the EGFR and ALK to detecting the cell blocks diagnosed as non-small cell lung cancer by the cell block immunocytochemistry.6.Statistical method: SPSS21.0 statistical software was used for data processing.Measurement data were tested by normality test and homogeneity test of variance.The data which did not conform to normal distribution were ranked by rank sum test.P<0.05 was statistically significant.The count data were checked with chi square test,if P<0.05 thought the difference was statistically significant.The diagnosis of diagnosed by histopathology and clinical is the gold standard.The receiver operating curve(ROC)is used to analyze the area under the curve(AUC).AUC>0.05 is considered to be of diagnostic value.Results: 1.The routine cytologic examination of exfoliated tumor cells in pleural effusion of the sensitivity,specificity and accuracy were 62.65%,87.01%,74.38%.DNA-image cytometry was used to detect the tumor cells in pleural effusion of the sensitivity,specificity and accuracy were 87.95%,93.51%,90.63%,were higher than that of conventional cytology.Through chi square after the test,the difference was statistically significant(P<0.05).Combined application of two methods can further improve the detection efficiency.The sensitivity,specificity and accuracy were 96.39%,85.71%,91.88%,were higher than that of conventional cytology.By chi square test,the difference was statistically significant(P<0.05).The area under the curve of routine cytology,DNA-image cytometry analysis and the combination of them was 0.7483,0.9073 and 0.9105,respectively.2.DNA-image cytometry analysis used the rank sum test to detect the number of DNA aneuploidy in different subtypes of lung cancer.The difference was not statistically significant(P>0.05).The positive rate detected in all subtypes of lung cancer was tested by chi square test,and the difference was not statistically significant(P>0.05).3.Make the "visible cancer"and "suspicious of cancer cells” of 55 cases of pleural effusion in exfoliated cells diagnosised by the routine cytologic diagnosis which have residual pleural effusion into the cell block.After continuous slice and HE staining,3 of 17 "suspected cancer cells" diagnosed by conventional cytology were diagnosed as adenocarcinoma and 1 were adenocarcinoma.After continuous cell sections,combined with immunocytochemical staining,9 cases of 17 cases of suspected cancer cells in routine cytology were diagnosed as adenocarcinoma,3 cases diagnosed as small cell carcinoma,and 5 cases diagnosed as mesothelial cells.At the same time,2 cases of metastatic adenocarcinoma cells can be diagnosed as lung metastasis of breast cancer.4.The cell blocks of 44 cases of non-small cell lung cancer patients diagnosed by cell block immunocytochemistry were detected by EGFR and ALK.15 cases(34.1%)were positive for EGFR,1cases(2.3%)were ALK positive,and the The expression of EGFR and ALK protein was detected in the tissue blocks of patients undergoing surgery or biopsy,and the conformance rate of tissue block and cell block EGFR and ALK protein expression was 100%.Conclusion: 1.The DNA-image cytometry analysis has important application value in the diagnosis of lung cancer.Combined with conventional cytology test,it can improve the detection rate of lung cancer and has important clinical value.2.The DNA-image cytometry analysis has no statistical significance in detecting the number and positive rate of DNA aneuploid cells in different subtypes of lung cancer.It has no help for lung cancer typing.3.The Cell block combined with immunocytochemical detection technology has important clinical value in judging the histologic origin of exfoliated cells in pleural effusion,and helps to diagnose the specific histological type of lung cancer.4.Conventional cellular methods,DNA-ICM methods and cell block immunocytochemistry can make up for each other's deficiencies,which can help the diagnosis,histological typing and further molecular typing of lung cancer.5.The protein expression of EGFR and ALK was detected after continuous slice of cell block,which was helpful for clinical targeted therapy.It is possible to provide a new source of specimens for subsequent EGFR and ALK gene detection in patients with advanced lung cancer who cannot obtain histological specimens. |
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