Clinical significance of expression of EGFR and the diagnostic value of cell wax block in malignant pleural effusions of lung adenocarcinoma

Objective To investigate the diagnostic value of detection of protein SP70 in differentiating benign and malignant pleural effusion. Methods A case-control study was conducted from July 2011 to February 2012. 108 cases of pleural effusion from patients with clinically proven lung cancers and 122 cases of benign pleural effusion were collected.SP70 was detected by Sandwich ELISA，while CEA，CYFRA21-1，NSE were measured by electrochemiluminescence immunoassay for comparison.Meanwhile，protein SP70 on exfoliated cells in pleural effusion was detected by direct immunofluorescence， and was compared with the results of HE staining.The differences between the groups were evaluated by the chi-square test Fisher′s exact test. Results Positive rates of SP70，CEA，CYFRA21-1，NSE were 72.2%，58.3%，52.8% and 30.6% in malignant pleural effusion，obviously higher than benign pleural effusio(9.8%，13.1%，23.0% and 19.7%).The specificity of SP70，CEA，CYFRA21-1，NSE were 90.2%，86.9%，77.0% and 80.3%，NSCLC had significantly higher positive rate than SCLC（74.3%>0.0%，P=0.02<0.05）， detection of protein SP70 in malignant pleural effusion had significantly higher coincidence rate than HE staining（72.2% vs 47.2%， χ2 =14.03，P<0.05）. Conclusion Determination of the protein SP70 in pleural effusion and in exfoliated cells，can improve the sensitivity and specificity of the diagnosis of malignant pleural effusion.（Chin J Lab Med，2012,35：1150-1154） Key words: Enzyme linked immunosorbent assay; Fluoroimmunoassay; Pleural effusion; Tumor markers，biological

Objective To disscuss the value of medical thoracoscopy in the diagnosis of malignant pleural effusion and probe the clinical significance of the diagnosis of malignant pleural effusion of the level of carcinoembryonic antigen (carcino-embryonic antigen, CEA) in pleural fluid, serum. Methods We analyzed retrospective the clinical data of 77 patients with pleural effusion during August 2015 to August 2016.We took the medical thoracoscope pathological results as the gold standard and evaluated the value of the level of CEA in pleural effusion and in serum and the pleural effusion/serum CEA ratio to diagnosis malignant pleural effusion. Results ①Among the 77 patients with pleural effusion, medical thoracoscope could clearly diagnosis 74 of them (46 of them were with malignant pleural effusion, 27 of them were with tuberculous pleural effusion, 3 of them were with pneumonia pleural effusion in cases, one was with nonspecific inflammation incase), the diagnostic rate was 96.1%.②The level of CEA in serum、in pleural effusion and the pleural effusion/serum CEA ratio in the malignant pleural effusion groupobviously were higher than that in pleural effusion group.Therefore, the level of CEA in pleural effusion of ladenocarcinoma were significantly higher than thoseother types of pleural effusion.③The sensitivity of the CEA in pleural effusion, specificity and accuracy ratio (80.4%, 89.3%, 83.7%) were higher than that in serum CEA and the pleural effusion/serum CEA ratio (56.5%, 78.5% and 64.9%, respectively, 93.4%, 64.2%, 82.4%). Conclusions ①Medical thoracoscopy is a safe, effective, practical diagnosis method which is worth to popularizing widely.②The CEA levels in the body, especially in pleural effusion, is significant to diagnosis the malignant pleural effusion(especially in adenocarcinoma). It can be used as an auxiliary diagnostic index for clinical application. Key words: Pleural effusion, malignant; Medical thoracoscopy; The CEA in serum; The CEA in pleural effusion; Pleural effusion/serum CEA ratio

Objective To investigate the value of osteopontin(OPN) in the differential diagnosis of pleural effusion.Methods A total of 85 patients with pleural effusion during October 2009 to January 2011 were recruited at the department of respiratory medicine of Hangzhou First Hospital.Pleural effusion was malignant in 39 cases and benign in 46 cases.The patients were divided into a malignant pleural effusion group,an infective pleural effusion group,and a transudation group.Then the infective was further divided into subgroup of tuberculosis pleural effusion group and parapneumonic effusion group.Results The concentration of OPN was significantly higher in the malignant pleural effusion group compared with the benign pleural effusion groups (P ＜70.01).When the cutoff value of OPN was set at 2 000 ng/L,the sensitivity specificity and accuracy for the diagnosis of malignant pleural effusion was 71.8％,95.7％ and 84.8％,respectively.Furthermore,the detection of OPN in combination with CEA showed better diagnostic sensitivity (97.43％),specificity (95.65％),and accuracy (96.47％).Conclusions The detection of OPN in pleural effusion is of some clinical significance in the differential diagnosis of malignant and benign pleural effusion. Key words: Osteopontin; Pleural effusion; Tumor markers; Lung cancer

Objective To investigate the value of paraffin-embedded section of cell block in the diagnosis of benign or malignant pleural and peritoneal effusion. Methods 105 patients suspected with malignant pleural and peritoneal effusion admitted into tour hospital from November, 2015 to December, 2017 were selected as the research objects. The specimens of pleural and peritoneal effusion of all the patients were collected. And these specimens were given paraffin-embedded section of cell block, cell smear, and exfoliative cell pathological examination. The results of exfoliative cell pathological examination were taken as the gold standard. The positive rates of malignant pleural and peritoneal effusion diagnosed by paraffin-embedded section of cell block and cell smear were evaluated. The diagnostic value of the two methods was compared. Results Among the 50 pleural effusion specimens, the results of exfoliative cell pathological examination showed that there were 36 cases of malignant pleural effusion. Among the 55 peritoneal effusion specimens, the results of exfoliative cell pathological examination showed that there were 42 cases of malignant peritoneal effusion. The positive detection rate of pleural effusion diagnosed by paraffin-embedded section of cell block (80.56%) was higher than that diagnosed by cell smear (58.33%), with a statistical difference (P<0.05). The positive detection rate of peritoneal effusion diagnosed by paraffin-embedded section of cell block (85.71%) was higher than that diagnosed by cell smear (64.29%), with a statistical difference (P<0.05). The sensitivity, specificity, and accuracy of paraffin-embedded section of cell block in the diagnosis of malignant pleural and peritoneal effusion were higher than those of cell smear, with statistical differences (all P<0.05). Conclusions Paraffin-embedded section of cell block in the differential diagnosis of benign or malignant pleural and peritoneal effusion has high sensitivity and specificity, and can increase the diagnostic accuracy and provide an important basis for clinical diagnosis and treatment of the disease. Key words: Pleural effusion; Paraffin-embedded section of cell block; Cell smear; Exfoliative cell pathological examination

To establish a method (negative enrichment by immunomagnetic beads) for detection of tumor cells in pleural effusions and to evaluate the sensitivity and specificity of the method for clinical application. Five, 10, 20, 50 and 100 A549 (lung adenocarcinoma) cells were labeled with DAPI and added into 20 ml pleural effusions [containing (1 - 10)×10(6)cells] from heart failure patients, followed by immunomagnetic negative enrichment method. Recovered cancer cells were enumerated using a fluorescent microscope. Tumor cells were enriched from pleural effusion samples by means of density gradient centrifugation and negative enrichment by immunomagnetic beads method, followed by identification with cytology analysis (Wright's Giemsa's staining), immunofluorescence staining (IF) and fluorescence in situ hybridization (FISH) using centromere DNA probes of chromosome 7 and 8. Cytology, IF and FISH evaluations were performed in 53 pleural effusion samples, including 36 cases of malignant disease (25 male and 11 female patients aging 40 to 78 years, mean age (63 ± 9) and 17 cases of benign disease (8 male and 9 female patients aging 25 to 81 years, mean age (53 ± 18). After DAPI staining and mixing with pleural effusions from heart failure patients, the cell recovery rates of A549 cells evaluated under fluorescence microscope were 75%, 78%, 82%, 85%, 88%, and the average recovery rate was 81.6%. Using negative enrichment method and density gradient centrifugation combined with cytology analysis, the positive rates of tumor cells in 36 malignant pleural effusion samples were 81% (29/36) and 61% (22/36), respectively (χ(2) = 4.00, P = 0.039). Using negative enrichment method combined with IF, the positive rate of CK18(+), DAPI(+), CD(45)(-) cells was 100%. Moreover, using negative enrichment method combined with FISH analysis, the positive rate of tumor cells was 86% (31/36), much higher than that using density gradient centrifugation combined with cytology analysis (χ(2) = 5.818, P = 0.012). In 17 cases of benign pleural effusions, using negative enrichment method combined with IF, the positive rate was 100%. But other methods didn't find cancer cells from benign pleural effusions. It was applicable to enrich tumor cells from pleural effusions using negative enrichment method by immunomagnetic beads. This method combined with cytology analysis or FISH significantly enhanced the sensitivity and specificity of tumor cell detection in pleural effusions. But it was difficult to distinguish cancer cells from mesothelial cells using immunofluorescence staining with CK18, DAPI and CD(45) label. More specific markers were needed to recognize tumor cells from pleural effusions.

Objective To study the levels and clinical significance of matrix metalloproteinase-9 (MMP-9) and endostatin in malignant pleural effusion of non-small cell lung cancer (NSCLC) patients.Methods MMP-9 and endostatin were detected in pleural effusion of 95 patients with NSCLC by ELISA method. Results The levels of MMP-9 and endostatin in pleural effusion of NSCLC patients were significantly higher than those in patients with benign pulmonary diseases (all P ＜0.01). The levels of MMP-9 and endostatin in pleural effusion of NSCLC patients were closely related to the histological classification ( P ＜0.05). By detection of MMP-9 and endostatin, the sensitivity was 67.9 ％ and 82. 1 ％,the specificity was 88. 1 ％ and 64.3 ％ for diagnosis of malignant pleural effusion. Conclusions MMP-9 and endostatin were significantly increased in pleural effusion of NSCLC patients. They had high sensitivity and specificity. Key words: Non-small cell lung cancer; Matrix metalloproteinase-9; Endostatin; Pleural effusion

Objective To evaluate the values of combined determination of CA72-4,CA242,CEA in the diagnosis of lung cancer pleural effusion. Methods Thirty-nine lung cancer pleural effusion samples and 35 benign pleural effusion samples were enrolled. The concentration of CA72-4, CA242, CEA in the pleural effusion of the 74 patients was determined with ELISA. The cut-off point for eath tumor marker was selected with receiver operating characteristic curves. The results were analyzed with sensitivity,specificity,positive prognostie value,negative prognostic value and accuracy. Results The values of CA72-4,CA242,CEA in the lung cancer pleural effusion were higher than those in the benign groups(P<0.05). The area under curve for CA72-4, CA242, CEA were 0. 703,0. 727,0. 804 respectivly. The sensitivity and specificity of CA72-4 were 64.1% and 71.4 % in lung cancer pleural effusion,68.4% and 71.4 %. The sensitivity and specificity of CA242 were 59.0% and 80.0% in lung cancer pleural effusion. The sensitivity and specificity of CA72-4+ CEA and CA72-4 + CA242 + CEA were 89.7% and 94.3%. Both were better than other combinations.Conclusions The determination of CA72-4,CA242,CEA in lung cancer pleural effusion are useful for the diagnosis of malignant pleural effusion associated with lung cancer. The combined determination of the three tumor markers can increase the sensitivity and the speeificity in the diagnosis of lung cancer pleural effusion. Key words: Pleural effusion; Tumor markers; Lung cancer; CA72-4; CA242

The development of the inflammatory processes in the pleural space may result in increased pleural vascular permeability leading to the accumulation of fluid enriched in proteins and the recruitment of cells into the pleural space.1 Although pleural effusion (PE) is common, very little information is available on the inflammatory and immune mechanisms that are involved in its development. In particular, it is unclear which cells and mediators are involved in the inflammatory processes and whether resident immunocompetent cells may orchestrate the development of an inflammatory response. Lymphocytic PEs refers to those in which lymphocytes account for more than 50% of the total leukocytes in PE. Lymphocytic effusions are commonly (>90% of cases) the result of malignancy and tuberculous pleurisy, but can also occur with rheumatic diseases, chronic postcoronary artery bypass effusion, chylothorax, acute lung allograft rejection and yellow nail syndrome.2 In normal humans a small amount of PE is present, but the exact volume of this fluid is unknown. In normal PE, there is a predominance of macrophages and lymphocytes; mesothelial cells, neutrophils and eosinophils are only marginally present.3 In an early study, it has been demonstrated that both the percentage and absolute numbers of T lymphocytes in tuberculous PE were significantly higher than in the corresponding peripheral blood. On the other hand, in patients with pulmonary tuberculosis, pulmonary malignancy or nonspecific pleurisy the percentages and absolute numbers of B lymphocytes were significantly lower in PE than in peripheral blood.4 It has been reported that PE is enriched with CD4+CDw29+ T cells, which are thought to represent “memory” T cells, and these PE CD4+CDw29+ cells, but not CD4+CDw29- cells, proliferated vigorously and produced high levels of interferon (IFN)-γ when stimulated with a purified protein derivative of Mycobacterium tuberculosis.5 Therefore, in tuberculous PE, CD4+CDw29+ cells are concentrated at the site of disease activity, produce IFN-γ and are likely to play an important role in the local human cell-mediated immune response to Mycobacterium tuberculosis. Malignant PE is frequently observed in lung cancer and a diagnosis of malignant PE with lung cancer carries a poor prognosis. In malignant PE, CD4+ T cells are dominant and the proportion of CD8+ T cells is significantly lower than that of CD4+ T cells.6 In contrast, the proportion of CD4+ T cells in the pleural cavity of patients with lung cancer without malignant PE is significantly lower than that of CD8+ T cells.7 Furthermore the proportion of PE CD4+ T cells may help to select patients who are likely to have a poorer prognosis after surgery and therefore may be suitable for consideration of adjuvant treatments.8 Immunoregulatory T cells have been believed to be involved in the control of the local immune response and in the growth of malignant tumors.9 Studies ongoing for more than a decade have provided firm evidence for the existence of a unique CD4+CD25+ T-cell population of “professional” regulatory/suppressor T cells that actively and dominantly prevent both the activation and the effector function of autoreactive T cells that have escaped other mechanisms of tolerance.10,11 Our previous study has shown that CD4+CD25+ T-cell numbers in malignant PE were much higher than those in PE from patients with lung cancer without malignant effusion and higher than numbers in peripheral blood. Our data also revealed that CD4+CD25+ T cells infiltrating PE were regulatory T cells as they express high levels of Foxp3 transcription factor. Moreover, pleural CD4+CD25+ T cells could potently suppress the proliferation of CD4+CD25- T cells and cytotoxic lymphocyte-associated antigen-4 was involved in the suppressive activity of pleural CD4+CD25+ T cells.12 In addition, we have demonstrated that CD4+CD25+ T cell numbers in tuberculous PE were much higher than those in peripheral blood from patients with tuberculous PE and from healthy subjects, and that the CD4+CD25+ T cells infiltrating pleural space were regulatory T cells as they expressed high levels of Foxp3 transcription factor. Moreover, tuberculous PE-derived CD4+CD25+ T cells could potently suppress the proliferation of responding T cells.13 It has been reported that the long-term effects of adoptively transferred CD4+CD25+ T cells induced ex vivo are due to their ability to generate new cytokine-producing CD4+CD25+ T cells in vivo.14 We speculated that an increased percentage of CD4+CD25+ T cells in PE might be due to either active recruitment or local differentiation. It has been demonstrated that human CD4+CD25+ T cells preferentially move to and accumulate in tumors and ascites, but rarely enter draining lymph nodes in later cancer stages.15 In a previous study, we provided direct evidence that interleukin (IL)-16 is capable of inducing CD4+ T-cell infiltration into the pleural space.16 Therefore, as a subpopulation of CD4+ T cells, CD4+CD25+ T cells might also be recruited into PE by local production of IL-16, because the IL-16 level is significantly higher in PE than in serum.16 Since we observed that CD4+CD25+ T cells were overrepresented in malignant and tuberculous PEs, this raised the question whether these cells could be recruited from the circulation. In a recent study, we have found that the level of chemokine CCL22 in PE correlated best with the numbers of CD4+CD25+ T cells, and that an anti-CCL22 antibody inhibited the ability of the PE to stimulate peripheral CD4+CD25+ T cell chemotaxis in vitro. Moreover, intrapleural administration of human recombinant CCL22, but not vehicle, into patients with PE produced a marked progressive influx of CD4+CD25+ T cells into the pleural space when studied by flow cytometry (our unpublished data). Our results demonstrated that CCL22 is produced by pleural cells and would appear to play a crucial role in the directed migration of CD4+CD25+ regulatory T cells into the pleural space. The role of T cells as active players in the mediation of pleural pathologies is beginning to be recognized. Understanding the exact roles of T cells in pleural space may provide a foundation for potential future attempts to augment lymphocyte responses. Further study of specific cellular responses may provide insight into disease pathogenesis and may offer unique opportunities in the diagnosis and management of lymphocytic PE. For example, since CD4+CD25+ T-cell numbers are increased in malignant PE, and these pleural CD4+CD25+ T cells can potently suppress the proliferation of CD4+CD25- T cells, it is very possible to develop novel immune-boosting strategies based on eliminating this regulatory cell population in patients with cancer. Also, more work is required to delineate the role of CD4+CD25+ T cells in tuberculous PE. Further studies should be directed at identifying the mediators and mechanisms involved in the immunoregulatory properties of pleural CD4+CD25+ T cells in patients with tuberculous PE.

Objective To explore the role of matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TlMP-1)in the pathogenesis of tuberculous and tumorous pleural effusion,and to assess the value of MMP-9,TIMP-1 and MMP-9/TIMP-1 in the diagnosis and differential diagnosis of these two kinds of pleural effusion.Methods MMP-9 and TIMP-1 in the pleural fluid were detected by enzyme-linked immunosorbent assay in 36 patients with tuberculous pleuritis,38 patients with malignant tumor and 14 patients with transudates.Results ①MMP-9,TIMP-1 and MMP-9/TIMP-1 in tuberculous pleural effusion were higher than those in malignant pleural effusion and transudates(all P＜0.05).MMP-9.TIMP-1 and MMP-9/TIMP-1 in malignant pleural effusion were higher than those in transudates(all P＜0.05).②MMp-9 and MMP-9/TIMP-1 in malignant pleural effusion with positive cytological findings were higher than those with negative cytological findings(all P＜0.05) ,while TIMP-1 was lower in the former than that in the latter( P ＜0.05). ③MMP-9 positively correlated with TIMP-I( r =0. 239, P =0. 025). MMP-9 and MMP-9/TIMP-1 positively correlated with lactate dehydrogenase, adenosine deaminase, protein, total white blood cells and lymphocyte ratio in pleural effusion(all P＜0.01),and negatively correlated with glucose and chloride in pleural effusion (all P＜(0.01). TIMP-1 positively correlated with lactate dehydrogenase, adenosine deaminase, protein and lymphocyte ratio in pleural effusion(all P＜0.01 ). ④In diagnosing tumorous pleural effusion, the sensitivities of MMP-9, TIMP-1 and MMP-9/TIMP-1 were respectively 63.2%, 71.1% and 65.8%, the specificities were respectively 83.3%, 63.9% and 80.6%. However, in the diagnosing process, the sensitivity and specificity were 39.5% and 91.7% when MMP-9 and TIMP-1 were combined in a series way, 94.70% and 55.6% in a parallel way.Conclusions MMP-9 and TIMP-1 are closely related to the tuberculous and tumorous pleural effusion. MMP-9/TIMP-1 plays a great role in the pathogenesis of tuberculous and tumorous pleural effusion. Detection of MMP-9, TIMP-1 and MMP-9/TIMP-1 in pleural effusion contributes to the differential diagnosis of tuberculous pleurisy and malignant pleural effusion. Key words: Lung tumor; Tuberculosis; Pleural effusion; Matrix metalloproteinase-9; Tissue inhibitor of metalloproteinase-1

Objective To determine whether pleural effusion samples, including cell-free DNA (cfDNA) samples and pleural effusion cancer cells, could be an alternative of tumor tissue samples to test epidermal growth factor receptor (EGFR) gene mutation status. Methods A total of 46 patients with lung adenocarcinoma diagnosed in the Department of Respiratory and Critical Care Medicine, Xiangya Hospital from September 2013 to January 2015 were included in this study.The matched pleural effusion cfDNA samples, cancer cells, tumor tissues samples were collected from these patients.EGFR mutation status in different kinds of samples was determined by liquid-chip platform. Results EGFR mutation was identified in 20 out of 46 (43.5%) pleural effusion cfDNA samples, 13 out of 36 (36.1%) pleural effusion cancer cell samples, and 21 out of 46 (45.7%) matched tumor tissues.The overall concordance of the EGFR gene mutation detected in pleural effusion cfDNA samples and tumor tissue samples was 93.5% (43/46, κ=0.868). The overall concordance of the EGFR gene mutation detected in cancer cell samples and tumor tissue samples was 91.7% (33/36, κ=0.822). Conclusions Pleural effusion cfDNA samples and cancer cell samples have high concordance with matched tumor tissue samples, suggesting that pleural fluid is an alternative for detecting EGFR mutation when the biopsy of tumor tissue is infeasible. Key words: Lung adenocarcinoma; Receptor, epidermal growth factor; Tumor; Pleural effusion; Cell-free DNA; Cancer cells

Objective To investigate the clinical application value of flow cytometry (FCM) established by specific fluorescent monoclonal antibody in detecting lung cancer cells in pleural effusion with non-small cell lung cancer(NSCLC). Methods The FCM assay was established by using fluorescent monoclonal antibody NJ001 which could specifically recognize NSCLC. A case-control study was conducted to evaluate the level of lung cancer cells in pleural effusion among 32 cases of NSCLC (including 29 lung adenocarcinoma cases and 3 lung squamous cell carcinoma cases) and 26 cases of benign disease with definite diagnosis collected from the First Affiliated Hospital of Nanjing Medical University from February 2014 to August 2014. The data of FCM assay between NSCLC and benign disease were evaluated by the rank sum test. The data were analyzed by ROC curve to assess the diagnosis value. Those FCM results among 32 cases of NSCLC were compared with Papanicolaou staining by the chi-square test. Meanwhile, NJ001 specific antigen on exfoliate cells was detected by direct immunofluorescence assay and comparing with FCM assay by the chi-square test among 22 cases of NSCLC. Results The number of positive events by FCM assay was 5(0-18)/100 000 in benign pleural effusion and 100(2-988)/100 000 in pleural effusion of NSCLC, which was significantly higher compared with the benign disease (Z=-5.647, P<0.001). The area under the ROC curve was 0.934 and the optimal diagnostic critical value was 18/100 000. This method performed a high sensitivity[87.5% (28/32)] and specificity[100.0%(26/26)]. The positive rate in 29 lung adenocarcinoma cases was significantly higher by FCM assay than Papanicolaou staining[93.1%(27/29) vs 58.6%(17/29), χ2=7.627, P=0.006], and in 12 negative Papanicolaou staining lung adenocarcinoma cases the positive rate of FCM assay was 11/12; In 3 lung squamous cell carcinoma cases the positive rate of FCM assay was 1/3 while all of Papanicolaou staining presented negative results. The FCM assay positive rate of 22 NSCLC cases was significantly higher than the direct immunofluorescence assay[90.9%(20/22) vs 59.1%(13/22), χ2=4.364, P=0.037]. Conclusion The FCM assay established by specific fluorescent monoclonal antibody applied for lung cancer cells especially for lung adenocarcinoma detection in pleural effusion had a high sensitivity and specificity, which was confirmed to be a valuable technological application for NSCLC clinical diagnosis. Key words: Carcinoma, non-small-cell lung; Pleural effusion; Flow cytometry

Objective To elucidate the significance of CYFRA21-1 and β2-microglobulin expression in pleural fluid and develop diagnostic application for malignant pleural effusion differentiation. Methods Detected expression of CYFRA21-1 by RIA analysis and β2-MG by immunoturbidimetry assay (ITA), in 40 cases of benign pleural effusion and 40 cases of malignant pleural effusion. Results The mean level of CYFRA21-1 in malignant pleural effusions was (78.80±24.90)μg/L higher than that in benign pleural effusions (26.20±15.60)μg/L (P<0.05) and β2-MG in benign pleural effusions was (6.11±0.87)mg/L, significantly higher than that in malignant pleural effusions which was (3.12±0.91)mg/L respectively (P<0.05). Conclusion Evaluation of CYFRA21-1 and β2-MG in chest fluid can improve the differential diagnosis of malignant pleural effusion from benign pleural effusion and significantly increase the sensitivity of diagnosis. Key words: Pleural effusion, malignant; Lung tumor; Cytokeration fragment antigen 21-1; β2-microglobulin

Objective To asses the diagnostic value of vascular endothelial growth factor (VEGF) in malignant pleural effusion by Meta-analysis. Methods We not only searched Best Evidence, Cochrane library, PubMed, CBM, CNKI , VIP, Wanfang Data et al but also checked online (www.google.cn; www.baidu.com) and traced literature to gather the relevant research literature on VEGF for the diagnosis of malignant pleural effusion from January 2000 to May 2016. Then we used Meta-Disc 1.4 and RevMan 5.3 software for data analysis to evaluate the diagnostic value of VEGF in malignant pleural effusion by calculating the pooled sensitivity, specificity, the likelihood ratio(LR), diagnostic odds ratios (DOR) and conducting summary receiver operating characteristic curve(SROC curve). Results Totally 747 articles were searched and finally 10 were included with sample size of 893 cases, including 468 cases of malignant pleural effusion and 425 cases of benign pleural effusion. The summary estimates for VEGF in the diagnosis of MPE were: sensitivity 0.72(95% CI 0.67, 0.76), specificity0.80 (95% CI 0.75, 0.83), positive likelihood ratio 3.81(95% CI 2.40, 6.06), negative likelihood ratio 0.37(95% CI 0.28, 0.48), diagnostic odds ratio 12.79(95% CI 6.33, 25.84) and the SROC area under the curve(AUC) 0.8384. Conclusions VEGF in pleural effusion has potential diagnostic value with advanced sensitivity and specificity and it can be used as adjunct tool for non-invasive diagnosis of malignant pleural effusion. Key words: Vascular endothelial growth factor; Pleural effusion; Diagnosis; Meta-analysis

Objective To investigate the expression and clinical significance of mineral dust-induced gene(MDIG) in malignant pleural effusions (MPE) and tuberculosis pleural effusions (TBPE).Methods Fifty-four patients with MPE (MPE group) and 50 patients with TBPE (TBPE group) were collected.The MDIG protein was detected by enzyme-linked immunosorbent assay and the relative quantification of MDIG mRNA was measured by real-time quantitation polymerase chain reaction.The cutoff value,sensitivity and specificity of the MDIG protein and MDIG mRNA to diagnose the MPE were evaluated by receiver operating characteristic (ROC) curve analysis.By Spearman correlation analysis,the correlation of MDIG protein and MDIG mRNA was evaluated.Results The MDIG protein in MPE group was significantly higher than that in TBPE group [(304.38 ± 228.47) ng/L vs.(44.43 ± 40.57) ng/L],and there was statistical difference (P ＜ 0.01).The MDIG mRNA in MPE group was significantly higher than that in TBPE group (6.27 ± 3.54 vs.1.82 ± 0.64),and there was statistical difference (P ＜ 0.01).With a cutoff point of 114.23 ng/L,MDIG protein had a sensitivity of 77.8％ and a specificity of 94.0％ for differential diagnosis.With a cutoff point of 2.75,MDIG mRNA had a sensitivity of 88.9％ and a specificity of 92.0％ for differential diagnosis.There was a positive correction between MDIG protein and MDIG mRNA (r =0.915,P ＜ 0.01).Conclusions The MDIG protein and MDIG mRNA are highly expressed in MPE with a good sensitivity and specificity.MDIG protein and MDIG mRNA maybe a good clinical indicator in the diagnosis of malignant pleural effusions. Key words: Tuberculosis; Pleural effusion, malignant; Mineral dust-induced gene

Objective To investigate the change of serum cancer antigen(CA) 125 in patients with pleural effusion.Methods One hundred and twenty-eight patients with pleural effusion were admitted to the Naval General Hospital of People's Liberation Army from January 2010 to September 2012 were selected as our subjects.The level of serum CA125 was measured.The difference of serum CA125 positive rate and level were compared according to gender,pleural effusion nature,quantity and pleural effusion chest area; And the difference of patients with malignant pleural effusion tuberculosis,inflammatory,exudative pleural effusion based on above indicators.The correlation between serum CA125 level and pleural effusion depth were analyzed.Results The positive proportions of CA125 were 83.3％ (35/42) and 76.7 ％ (66/88) of patients with malignant and benign effusion respectively,and there was no significant difference (x2 =0.74,P ＞ 0.05).The serum CA125 level of patients with malignant pleural effusion was significantly higher than benign ones ((177.8 ± 31.4) U/ml vs.(110.6 ± 13.6) U/ml,t =31.24,P ＜ 0.05).There were no significant difference in the positive proportion of serum CA125 between malignant,tuberculous,inflammatory and transudative pleural effusion(75.8％ (25/33),70.0％ (20/29),87.5％ (21/24) and 83.3％ (35/42),P ＞ 0.05).Serum CA125 levels of patients with malignant pleural effusion were significantly higher than that with inflammatory ((177.8 ± 31.4) U/ml vs.(72.5 ± 12.8) U/ml,P ＜ 0.05),but the differences were not significant among malignant,tuberculous and transudative pleural effusion group((140.6 ± 28.2) U/ml,(154.3 ± 30.5) U/ml,P ＞ 0.05).The serum CA125 levels of patients with small,moderate and large effusions were (56.4 ± 18.2) U/ml,(120.2 ± 24.5) U/ml and (185.5 ± 34.6) U/ml respectively,and the difference among these groups were significant(F =296.03,P ＜ 0.05).Serum CA125 levels was positively correlated with pleural fluid depth (r =0.56,P ＜0.01).Different gender,pleural effusion parts serum CA125 positive rate and the different levels were not statistically significant (P ＞ 0.05).Conclusion Serum CA125 increased in patients with benign and malignant pleural effusion,and serum CA125 was not severed as the diagnosis biomarker in differentiating benign and malignant pleural effusion.Serum CA125 levels is helpful in monitoring the change of pleural fluid size due to the relation with depth of pleural fluid. Key words: Pleural effusion; Cancer antigen 125 ; Serum