| Pleural fluids are common complications that may be produced by a wide ranging of diseases especially lung cancer. It is important medium to elucidate the precise etiology of the pleural fluid especially to discriminate benign from malignant fluids. Routine cytology examinations are mainly based on the morphologic characteristic of cells, but its sensitivity is lower for that there are also the morphologic characteristic of cancer cells for stimulated reactive mesothelial cells from the serous membrane, a considerable number of false negative or positive results were founded in the diagnosis evenly. Tumor markers are defined as biomolecules that can be measured quantitatively in body tissues or fluids and some can be clinically useful for differentiation malignant from benign tumors. It is founded to be one of ideal method for diagnosing lung cancer by detecting the tumor markers in the pleural fluids recently. There were much mucus, blood, inflammatory cells and necrotic material in the pick - and - smear slides, resulting in a lower detection rate. Liquid - based Cytologic Test (LCT) has been applied for cervical cytology diagnosis successfully and widely, however it is not reported for sputum cytology diagnosis in the world at present. The aim of this study was to evaluate the practical value in clinic of tumor markers and LCT in the pleural fluid and the sputum of patients with lung cancer respectively.Methods1. Subjects investigatedA total of 101 pleural fluid samples collected from the patients in the outpatient and inpatient of First affiliated hospital, China Medical University were in-eluded in this study from March , 2003 to February, 2004. There were 56 males and 45 females, the age is from 24 to 90. The pleural fluid of patients with lung cancer is including of squamous cell carcinoma ( SCC) 7, adenocarcinoma ( A-DEN) 56 and small cell lung carcinoma (SCLC) 3; the one of patients without lung cancer involving of inflammation 20, tuberculosis 10, lymphoma 5. All of them were confirmed by the histopathology and the cytology of bronchoscope.Another 708 patients with respiratory symptoms were collected in the outpatient and inpatient of First affiliated hospital, China Medical University, 101 of whom diagnosed by the histopathology and the cytology of bronchoscope as lung cancer were included in this study from January to Octuber, 2004. There were 68 males and 33 females, the age is from 30 to 93, the histologic classified is including of squamous cell carcinoma (SCC) 48, adenocarcinoma (ADEN) 36 and small cell lung carcinoma (SCLC) 17.2. Specimen preparationSupernatant of the pleural fluid obtained by centrifugation at 10000 g for 30 min were tested by chemiluminescent enzyme immunometric assay and western blotting, the 4 slides smeared from sediment were examined by routine stain of Pap.82 sediments selected were prepared for the concentration of cell about lOV L to be detected by cytochip, including of adenocarcinoma (ADEN) 56, squamous cell carcinoma (SCC) 6, inflammation 13, tuberculosis 7.51 sediments selected were prepared for transmission electron microscope examination, including of adenocarcinoma ( ADEN ) 41, inflammation 7, tuberculosis 3.The sputum were poured into the little bottles with CytoRich, the volume of sputum was the same as the one of CytoRich. Then add the mucolytic agents per 1 ml to the 10 ml mixed liquor above named, the mixed liquor were subjected to it for 3o min, jarred for 10 seconds. We also can add some mucolytic agents to the mixed liquor again if concrete was founded in the mixed liquor.3. ReagentsProtein quantity: Carcinoembryonic antigen ( CEA) , Carcinoma antigen 153(CA153) , Cytokaretin 19 segment (CYFRA 21 -1) , produced DPC com-pany in U. S. A. , bought Tianjin Depu biotechnology and medical products corporation.Western blotting: CEA, bought Sino -American Biotechnology Co..Cytochip: Epithelial specific antigen (ESA) , CD44V6, ND - 1, CD3, CD45RO, CD20, CD79a, CD15, CD30, CD68, All of them were monoclonal antibody. ND -1 was developed by Song jin - dan and so on, the department of cell biology, residual were bought Maixin - Bio in Fu Zhou.LCT: CytoRich and mucolytic agents were produced Tripath company in U. S. A. , bought Hongkong xinshun limited corporation.4. MethodsProtein quantity: 101 pleural fluid and 66 serum of patients with lung cancer were analyzed by IMMULITE made in DPC company in U. S. A.Western blotting: Concentration determination of protein, irrigation page, preparation examples, electrophoresis, transfer membrane, stain, hybridize, ECL reaction and washing film.Cytochip: spot sample, block out, hybridize, fixation and stain.TEM: fixation, post fixation, dehydrate, embed, slice up, double stain and microscopic examination.LCT: mucolyic, centrifugation, vibrate, preparation slides and microscopic examination.5. Statistical Analysis: t tests and x2 tests by SPSS 10.0.ResultsThe levels of protein quantities from CEA, CA153 and CYFRA21 - 1 and the levels of western blotting from CEA in the pleural fluid of patients with lung cancer were significantly higher than those of patients without lung cancer ( P < 0.01 ) ; The levels of 4 markers in the pleural fluid were obviously higher than in the serum of patients with lung cancer ( P < 0.01) ; Western blotting was obviously superior to protein quantity about CEA, the sensitivity and the specificity were 84. 9% and 91. 4%, respectively; The positive numbers of ESA, CD44V6, ND -1 were 52, 45, 56 respectively by cytochip hybridization in the pleural fluids of patients with 62 lung cancers, the point of positive hybridization...
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