Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Pleural Effusion, Malignant ; Consensus ; China

OBJECTIVESTo detect CEA mRNA levels in benign and malignant pleural and peritoneal effusions and evaluate their clinical significance.

METHODSSamples of pleural and peritoneal effusions from 58 patients with malignant diseases and 76 patients with benign diseases were collected and total RNAs were prepared and subjected to real-time fluorescent quantitative RT-PCR to determine the CEA mRNA levels in these samples. The positive rate of this examination was compared with that of shed cell pathological examination.

RESULTSNineteen samples (32.8%) of pleural and peritoneal effusions from the 58 patients with malignant diseases showed positive results in shed cell examination, while the number of CEA mRNA >1 CN was 46 (79.3%) (chi(2) = 21.81, P = 0.000). Nineteen samples of pleural and peritoneal effusions from the 76 patients with benign diseases showed CEA mRNA > 1 CN (25.0%), which was significantly different from that of the patients with malignant diseases (chi(2) = 38.85, P = 0.000).

CONCLUSIONCEA mRNA levels in pleural and peritoneal effusions can be quantified by real-time fluorescent quantitative PCR, which is more sensitive than shed cell pathological examination. This technique is helpful in discrimination of benign and malignant pleural and peritoneal effusions.

Ascitic Fluid ; chemistry ; Carcinoembryonic Antigen ; genetics ; Female ; Fluorescence ; Humans ; Male ; Middle Aged ; Pleural Effusion ; chemistry ; Pleural Effusion, Malignant ; chemistry ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; methods

BACKGROUND: Malignant pleural effusion is one of the common clinical manifestations of patients with lung adenocarcinoma. Patients with pleural effusion at the initial diagnosis of lung adenocarcinoma usually indicate poor prognosis. Epidermal growth factor receptor (EGFR) mutations mainly occur in patients with lung adenocarcinoma. Patients with different mutant subtypes have different prognosis. The clinical characteristics and prognostic factors of patients with EGFR mutated lung adenocarcinoma of different molecular subtypes combined with pleural effusion at initial diagnosis are still unclear. This study was designed to explore the clinical characteristics and prognostic factors of these patients in order to provide management recommendations for them. METHODS: A retrospective analysis of the clinical characteristics, treatment, outcomes and progression-free survival (PFS) of first-line treatment in patients with EGFR mutated lung adenocarcinoma combined with pleural effusion at initial diagnosis admitted to Department of Medical Oncology and Radiation Sickness, Peking University Third Hospital from January 2012 to June 2021 was performed. Pearson's chi-square test or Fisher's exact test were performed for comparison between groups. Kaplan-Meier method was performed for survival analysis and Cox proportional risk regression model was performed for multivariate analysis. RESULTS: 76 patients met the inclusion criteria in this study. The incidences of EGFR classical mutations 19del, 21L858R and non-classical mutations were 46.0%, 38.2% and 15.8%, respectively among these patients. There was no significant difference between the three mutations in terms of gender, age, presence of dyspnea at presentation, whether other distant metastases were combined, site of pleural effusion, volume of pleural effusion, presence of other combined effusions, tumor-node-metastasis (TNM) stage, presence of other gene mutations, and treatment of pleural effusion (P>0.05). In patients with EGFR classical mutations 19del or 21L858R or non-classical mutations subtype, the proportion of chemotherapy in first-line regimens were 17.1%, 20.7% and 58.3%, respectively (P=0.001); and first-line disease control rates were 94.3%, 75.9% and 50%, respectively (P=0.003); pleural effusion control rates were 94.3%, 79.3% and 66.7%, respectively (P=0.04); PFS were 287 d, 327 d and 55 d, respectively (P=0.001). Univariate analysis showed that EGFR mutation subtype, control of pleural effusion, first-line treatment agents, and first-line treatment efficacy were significantly associated with PFS (P<0.05). Cox multifactorial analysis showed that only EGFR mutation subtype and first-line treatment efficacy were independent prognostic factors for PFS (P<0.05). CONCLUSIONS PFS was significantly better for classical mutations than for non-classical mutations in patients with EGFR mutated lung adenocarcinoma combined with pleural effusion at initial diagnosis. Improving the efficacy of first-line therapy is the key to improve the prognosis of these patients.
Adenocarcinoma of Lung/genetics* ; ErbB Receptors/genetics* ; Humans ; Lung Neoplasms/pathology* ; Mutation ; Pleural Effusion/complications* ; Prognosis ; Retrospective Studies

Pericentric inversion of chromosome 4 can give rise to 2 alternate recombinant (rec) chromosomesby duplication or deletion of 4p. The deletion of distal 4p manifests as Wolf-Hirschhorn syndrome (WHS). Here, we report the molecular cytogenetic findings and clinical manifestations observed in an infant with 46,XX,rec(4)dup(4q)inv(4)(p16q31.3)pat. The infant was delivered by Cesarean section at the 33rd week of gestation because pleural effusion and polyhydramnios were detected on ultrasonography. At birth, the infant showed no malformation or dysfunction, except for a preauricular skin tag. Array comparative genomic hybridization analysis of neonatal peripheral blood samples showed a gain of 38 Mb on 4q31.3-qter and a loss of 3 Mb on 4p16.3, and these results were consistent with WHS. At the last follow-up at 8 months of age (corrected age, 6 months), the infant had not achieved complete head control.
*Chromosome Deletion ; *Chromosome Duplication ; *Chromosome Inversion ; *Chromosomes, Human, Pair 4 ; Comparative Genomic Hybridization ; Female ; Gestational Age ; Humans ; Infant ; Pleural Effusion/ultrasonography ; Polyhydramnios/ultrasonography ; Pregnancy ; Wolf-Hirschhorn Syndrome/*genetics

Isolated pleural effusion, so called primary pleural effusion denotes a pleural effusion without documented etiology such as a cardiac, inflammatory, iatrogenic problem or fetal hydrops. Chromosomal anomaly such as Down syndrome may be associated with isolated pleural effusion. The content of the isolated pleural effusion is mostly chylous, and isolated non-chylous pleural effusion in neonate is rare. We experienced 2 cases of isolated non-chylous pleural effusion. They had neither cardiac problem nor other sign of hydrops fetalis. Imaging diagnosis was done by plain chest radiography and subsequent ultrasonogram. One of them was diagnosed to Down syndrome by karyotyping. They were fared well after diagnostic and therapeutic thoracentesis. We describe 2 cases of non-chylous pleural effusion and review a few English-language case reports of this entity.
Chyloperitoneum/pathology ; Chylothorax/pathology ; Down Syndrome/diagnosis/genetics ; Female ; Fetal Diseases/diagnosis/therapy ; Gestational Age ; Human ; Hydrothorax ; Infant, Newborn ; Karyotyping ; Male ; *Pleural Effusion ; Pregnancy ; Ultrasonography ; Ultrasonography, Prenatal

BACKGROUNDThe discovery of water channel aquaporins (AQPs) has greatly expanded the understanding of the regulation of the water permeability of biological membranes. Aquaporin-1 (AQP1) may be involved in fluid transport in numerous pathological conditions. The objective of the present study was to examine whether AQP1 is present in cultured rat pleural mesothelial cells (PMCs) and to investigate the specific inhibitory effect of RNA interference (RNAi) on AQP1 expression in PMCs, which may provide a new method for the further studies on the relation between expression of AQP1 in PMCs and pleural fluid removal in vivo.

METHODSPMCs were isolated and cultured from rat pleura. The expression of AQP1 in PMCs was confirmed by immunocytochemical staining and reverse transcriptase-polymerase chain reaction (RT-PCR). Two eukaryotic expression plasmid vectors of short hairpin RNA (shRNA) specific for the AQP1 gene of rat sapien were designed and constructed. The recombinant plasmid vectors were transfected into cultured rat PMCs by cation liposomes. Flow cytometry was used to screen the most effective shRNA at 48 hours after transfection. The expressions of AQP1 mRNA and protein were detected by RT-PCR and Western blotting method at 48 hours after transfection.

RESULTSRT-PCR and immunostaining revealed that AQP1 mRNA and protein were present in cultured rat PMCs. Two effective eukaryotic expression plasmid vectors of shRNA specific for the AQP1 gene were constructed successfully. The levels of the expression of AQP1 were inhibited by 83.45%, 90.93%, respectively, at mRNA level and 41.24%, 67.60%, respectively at protein level by two recombinant plasmids at 48 hours after transfection. The expression of AQP1 in PMCs transfected with plasmid was significantly lower than that of the cells transfected with the control plasmid HK and that of the untransfected cells (P < 0.01). There was no significant difference in AQP1 expression between the control group and the group transfected with AQP1 nonspecific shRNAs (P < 0.05).

CONCLUSIONSThe expression of AQP1 was present in rat PMCs. The application of shRNA-AQP1 could markedly inhibit the expression of AQP1 in cultured rat PMCs. The use of RNAi is a promising tool for future research into the mechanisms of pleural fluid in vivo.

Animals ; Aquaporin 1 ; antagonists & inhibitors ; genetics ; Cells, Cultured ; Epithelial Cells ; metabolism ; Flow Cytometry ; Male ; Microscopy, Fluorescence ; Pleura ; cytology ; metabolism ; Pleural Effusion ; therapy ; RNA Interference ; RNA, Small Interfering ; genetics ; Rats

OBJECTIVETo investigate aberrant methylation in the promoter of p16 gene in the sediment cells of pleural effusion and evaluate its clinical significance in the differentiating benign and malignant pleural effusion.

METHODSUsing methylation-specific PCR (MSP), aberrant promoter methylation of p16 gene was detected in the sedimental cells of pleural effusion samples from 66 patients with pleural effusion.

RESULTSOf the 66 patients with pleural effusion, 36 had a definite diagnosis of malignant pleural effusion, and the rest were confirmed to have benign pleural effusion. The positivity rate of p16 gene promoter methylation was 69.4% (25/36) in malignant pleural effusion and 13.3% (4/30) in benign pleural effusion specimens, showing a significant difference between them (χ² = 20.915, P < 0.01). The diagnostic sensitivity, specificity and accuracy of aberrant promoter methylation of p16 gene in the 36 malignant cases were 69.4%, 86.7% and 77.3%, respectively. The positive expression of p16 gene promoter methylation in malignant pleural effusion was not correlated to the histological type or the pathological grade of the tumor (P > 0.05).

CONCLUSIONDetection of aberrant methylation in p16 gene promoter in the sediment cells of pleural effusion specimens by MSP method allows differentiation between benign and malignant pleural effusion.

Adult ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Genes, p16 ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Pleural Effusion, Malignant ; diagnosis ; genetics ; Promoter Regions, Genetic ; genetics ; Sensitivity and Specificity

BACKGROUNDInterleukin (IL)-27 has been reported to have anti-proliferate and anti-angiogenic activities on cancer cells. However, the involvement of IL-27 in malignant pleural effusion (MPE) remains unknown. Thus, in this research, we compared the immune functions of IL-27, interferon (IFN)-γ, and IL-17 on lung cancer cells and revealed the regulatory mechanism of IL-27 in MPE.

METHODSThe distribution of IL-27 in both MPE and blood was evaluated by enzyme-linked immunosorbent assay and flow cytometry. The expressions of cytokine receptors and the levels of the phosphorylated signal transducer and activator of transcription (STAT) signalings were detected by flow cytometry. As well as the effects of proliferation, apoptosis, migration, and adherent activity of IL-27, IFN-γ, and IL-17 on lung cancer cells were also explored.

RESULTSThe expression of IL-27 was increased in MPE when compared with blood (147.3 ± 25.1 pg/ml vs. 100.3 ± 13.9 pg/ml, P = 0.04). IL-27 was noted to suppress both proliferation (18.33 ± 0.21 vs. 27.77 ± 0.88, P = 0.0005) and migration (1.82 ± 0.44 vs. 3.13 ± 0.07, P = 0.04) of A549 cells, but obviously promoted apoptosis of A549 cells (9.47 ± 1.14 vs. 4.96 ± 0.17, P = 0.02) by activating STAT1 signaling. Interestingly, IL-27 played totally opposite effects on A549 cells by activating STAT3 pathway. Moreover, IL-27 exerted different intercellular adherent activities of A549 cells to pleural mesothelial cell monolayer by activating different STAT signalings.

CONCLUSIONSIL-27 might exert an important immune regulation on lung cancer cells in human pleural malignant environment.

Adult ; Aged ; Aged, 80 and over ; Cell Line ; Cell Movement ; genetics ; physiology ; Cell Proliferation ; genetics ; physiology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interleukins ; metabolism ; Lung Neoplasms ; metabolism ; Male ; Middle Aged ; Pleural Effusion, Malignant ; metabolism ; Signal Transduction

OBJECTIVES: The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section. METHODS: A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was. RESULTS: Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21. CONCLUSIONS The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.
Carcinoma, Non-Small-Cell Lung/drug therapy* ; DNA Mutational Analysis/methods* ; ErbB Receptors/genetics* ; Female ; Humans ; Lung Neoplasms/drug therapy* ; Male ; Mutation ; Paraffin/therapeutic use* ; Pleural Effusion/genetics* ; Protein Kinase Inhibitors/therapeutic use* ; Staining and Labeling

OBJECTIVETo explore the mRNA expression of breast cancer susceptibility gene 1 (BRCA1) in tumor cells isolated from malignant pleural and peritoneal effusions, and the predictive role of BRCA1 related to the efficacy of cisplatin-based chemotherapy.

METHODSTumor cells were isolated from malignant pleural and peritoneal effusions of 31 cancer patients. The response of these tumor cells to cisplatin was determined by CCK8 assay. Real time quantitative RT-PCR was used to examine the BRCA1 mRNA level in the primary culture cancer cells.

RESULTSThe expression level of BRCA1 mRNA was 0.618 (0.014 - 18.063) in primary culture tumor cells. The IC(50) of DDP was 2.809 µg/ml in the primary culture tumor cells (0.118 - 19.439 µg/ml). Both BRCA1 mRNA expression and the tumor cells IC(50) of DDP were not significantly related with patient age, gender, the type of primary tumor, whether to accept the chemotherapy and effusion type (P > 0.05). The level of BRCA1 mRNA was negatively correlated with the chemosensitivity in terms of IC(50) of cisplatin (P < 0.001).

CONCLUSIONAssessment of expression level of BRCA1 mRNA may be useful in predicting the efficacy of cisplatin-based chemotherapy in patients with metastatic malignant effusions.

Antineoplastic Agents ; pharmacology ; Ascitic Fluid ; metabolism ; pathology ; BRCA1 Protein ; genetics ; metabolism ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Pleural Effusion, Malignant ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; metabolism ; pathology