Management of cytological material,pre‐analytical procedures and bio‐banking in effusion cytopathology

Serous effusion fluid is one of the most commonly encountered specimens in routine cytopathology practice. It provides invaluable information about the patient and the clinical status; but to get the most of it, specimen handling and processing must be carried out properly. Cytomorphology is the basis of a successful analysis which should complemented by ancillary tests when needed. A wide spectrum of ancillary techniques ‐ ranging from immunocytochemistry and flow cytometry to different assays of molecular pathology ‐ can be applied to serous effusions. This article describes the acquisition and management of serous effusion fluids, methods for preservation and transportation, different techniques of cytopreparation, application of immunocytochemistry, flow cytometry, and fluorescence in‐situ hybridization (FISH), as well as DNA extraction for polymerase chain reaction (PCR) and next generation sequencing (NGS). Principles of bio‐banking of effusion samples are also discussed which is getting more important in correlation with the developments in personalized medicine.     

恶性胸腔积液的治疗现状

恶性胸液在临床上基本上都出现在肿瘤的中、晚期,治疗的主要目的是解决患者的痛苦、提高病人的生存质量、延长患者的生命,其治疗措施随着科技的进步也在不断地改进完善,远期效果将有待于进一步提高。     

Characterization of IL-2-activated TILs and their use in intrapericardial immunotherapy in malignant pericardial effusion

Pericardial effusion (PE) and cardiac tamponade caused by malignant pericarditis are critical conditions in cancer patients, which still lack a recommended protocol for their long-term management. Percutaneous pericardiocentesis and simple drainage are commonly performed as the initial treatment. The aims of this study were to investigate the presence of cytotoxic T lymphocytes (CTLs) in malignant PE and to determine the clinical response to administering autologous tumor-infiltrating lymphocytes (TILs) into the pericardial cavity. Initially, we identified human lymphocyte antigen class-I-restricted and tumor-specific CTLs within the interleukin-2 (IL-2)-activated TILs in PEs from four patients, on the basis of interferon-γ production and lactate dehydrogenase-release assays. Clinically we observed favorable responses to the pericardial transfer of IL-2-activated autologous TILs in four patients: one male with advanced esophageal cancer, one female with recurrent lung cancer and two females with recurrent breast cancer, respectively. Autologous TILs from PEs were expanded in vitro with IL-2, characterized for CD3, CD4 and CD8 markers, checked for contamination and then infused into the patient’s pericardial space through a catheter. This was repeated biweekly. After treatment, there were no signs of recurrence of PE in either case, as determined by radiography, echocardiography and computed tomography. The only adverse effects seen were grade 1 fevers. These results suggested that intrapericardial cellular immunotherapy with autologous TILs could be a safe and effective treatment for controlling malignant pericarditis with associated cardiac tamponade, and that tumor-specific CTLs present in malignant PE might be important for tumor rejection.     

肺癌性胸腔积液中生存素基因检测的诊断意义探讨

目的：生存素基因（survivin）是一种新近发现的抗凋亡基因,在肿瘤组织中呈现表达。本文旨在探讨和比较肺癌性胸腔积液和结核性胸腔积液中生存素基因的表达情况,以及其联合细胞学检查对判断肺癌性胸腔积液的敏感度。方法：应用逆转录酶-聚合酶链反应法（RT-PCR）检测2007年06月～2008年03月42例肺癌患者癌性胸腔积液标本,及同时期28例结核性胸腔积液标本的生存素mRNA表达情况,并联合细胞学检查结果进行对比分析。结果：42例肺癌患者胸腔积液标本中生存素mRNA的阳性率为52138％（22／42）;癌细胞的检出率为30．95％（13／42）;生存素mRNA检测联合细胞学检查诊断肺癌的敏感性为61．90％（26／42）,显著高于单独胸腔积液细胞学检测的敏感性（P〈0．001）。28例结核性胸腔积液标本的生存素mRNA阳性率为7．14％（2／28）,显著低于肺癌患者胸腔积液标本生存素mRNA的阳性率（P〈0．001）。结论：运用RT—PCR方法检测胸腔积液中生存素mRNA的表达在判断肺癌性胸腔积液中具有一定的敏感性和特异性,可能作为肺癌辅助诊断的一个新检测指标。     

Immunological analysis and clinical effects of intraabdominal and intrapleural injection of lentinan for malignant ascites and pleural effusion

Twenty effusions in sixteen patients with malignant peritoneal and/or pleural effusions were treated with intracavitary injection of lentinan. Lentinan was injected at a dosage of 4 mg/week for 4 weeks. In total, sixteen (80%) of twenty lesions demonstrated clinical responses. Performance status was improved in seven patients. The average survival time in responders was 129 days, while, in non-responders, it was 49 days. Serious toxicities were not observed.NK activity of PBMC significantly decreased after lentinan injection. NK activity of PEC in responders was augmented significantly. Anti-Daudi and lymphokine activated killer activity were also augmented or maintained after lentinan injection.     

Examination of cytological smears and clot sections prepared from pleural fluids: a comparative study

Between January 1989 and May 1990, the Department of Cytology of Llandough Hospital examined 784 pleural fluid cytological preparations, 99 of which were also examined as paraffin wax sections of clots that had formed. to assess whether this practice yields worthwhile information, we reviewed the cytology and clot sections of 99 patients with pleural effusions and correlated the results with subsequent histology and review of clinical case notes. the specificities of each method separately and when combined was high: 92% clot section, 85% smear cytology and 85% combined. the sensitivity using clot section alone was 49%, smear cytology alone 46% and in combination, 54%. Analysis showed no significant difference between these sensitivities. When pleural fluid clot formation occurs, we recommend the continued examination of clot sections, since it yields an increase in positive diagnoses, albeit not statistically significant. Entre janvier 1989 et mai 1990, 784 préparations cytologiques de liquides pleuraux ont été examinées dans le département de Cytologie de l'Hôpital Llandough. Parmi ces 784 cas, 99 contenaient un coagulum qui a fait l'objet également de coupes histologiques apres inclusion en paraffine. Afin de déterminer si cette pratique apporte une information intéressante, la cytologie de ces 99 patients et les coupes obtenues á partir du coagulum ont été revues, puis les résultats one été corrélés avec les résultats histologiques et avec I'analyse des données cliniques. Les valeurs de la spécificité de chacune des méthodes et des deux méthodes associées sont élevées: 92% pout les coupes du coagulum, 85% pour la cytologie d'étalement et 85% pour les deux méthodes combinées. Les valeurs pour la sensibilité sont de 49% pour les coupes de coagulum, de 46% pour la cytologie seule et de 54% en combinant les deux methodes. L'analyse ne met pas en évidence de différence significativé de sensibilitk entre les méthodes. Lorsqu'un coagulum s'est formé dans un liquide pleural, nous recornmandons de poursuivre la pratique de l'inclusion en paraffine et de coupe de ce coagulum puisque cette pratique aboutit à une augmentation du taux de diagnostics positifs, même si la différence n'est pas démontrée statistiquement. Das zytologische Labor des Llandough Hospitals untersuchte zwischen Januar 1989 und Mai 1990 784 Pleurapunktate. Von 99 wurden ausserdem paraffinge-schnittone Clotpräparate angefertigt. Um den Wert dieses Verfahrens zu überprüfen wurden diese Ergebnisse mit späteren histologischen Proben und der Patientengeschichte korreliert. Die Spezifitaten betrugen: 92% für Clot, 85% für Ausstriche und 85% für die Kombination. Die Sensitivitäten: 49% für Clot, 46% für Ausstriche und 54% für die Kombination. Die Unterschiede sind statistisch nicht signifikant, trotzdem sollten Gerinnsel bei Vorhandensein histologisch verarbeitet werden.     

EGFR在晚期肺腺癌患者中的表达及临床意义

目的探讨表皮生长因子受体(EGFR)在晚期肺腺癌患者中的表达及其意义。方法收集60例晚期肺腺癌患者的胸腔积液及活检肺癌组织,采用免疫组化检测EGFR的表达,并探讨其表达与临床病理特征的关系。结果 EGFR在胸腔积液及肺癌组织中的阳性表达率分别为75.0%(45/60)、63.3%(38/60),两者差异无统计学意义(P0.05)。结论 EGFR均高表达于肺腺癌胸腔积液及腺癌组织,且与年龄、性别、吸烟史、分化程度及肿瘤的大小无明显相关,可指导肺腺癌患者的靶向治疗。     

血清和胸水中CA125在结核性和癌性胸水中的表达及临床意义

目的:探讨血清和胸水中CA125在结核性和癌性胸水中的表达及鉴别诊断意义。方法:抽选我院确诊的结核性胸水病人85例(结核组)和癌性胸水病人71例(癌症组),检测两组患者血清和胸水中CA125表达,并以胸水/血清中CA125比值10(p-CA125/s-CA12510)为临界值,观察其对癌性胸水的鉴别特异度、灵敏度及准确性。结果:癌症组胸水中CA125表达及p-CA125/s-CA125比值均显著高于结核组(P0.05);但血清中两组CA125表达比较差异无显著性(P0.05);两组胸水中,以35U/ml为临界值,两组患者阳性率92.9%(79/85)、100%(71/71)比较差异无显著性(X2=7.0718,P=0.0078)。癌症组中p-CA125/sCA125比值10的比率(84.5%VS 17.6%)明显高于结核组(X2=66.6244,P=0.0000);并以其为诊断癌性胸水的临界值,鉴别诊断特异度、灵敏度及准确性分别为82.3%、84.5%、83.3%。结论:血清和胸水中CA125表达对于鉴别结核性或者是癌性胸水的临床意义不大,但是p-CA125/s-CA125比值对于鉴别结核性和癌性胸水具有一定临床价值。