Long‐term cytological and histological outcomes in women managed with loop excision treatment under local anaesthetic for high‐grade cervical intraepithelial neoplasia

Y. L. Woo, C. Badley, E. Jackson and R. Crawford Long‐term cytological and histological outcomes in women managed with loop excision treatment under local anaesthetic for high‐grade cervical intraepithelial neoplasia Objective: This study examines the impact of excision margin status after large loop excision of the transformation zone (LLETZ) under local anaesthetic for high‐grade cervical intraepithelial neoplasia (HG‐CIN) on the cytological and histological outcomes up to 5 years after treatment. Methods: Prospective cytological and histological data were obtained by examination of the colposcopy database at Addenbrooke’s Hospital, Cambridge, UK. All women aged between 19 and 50 years who underwent treatment for HG‐CIN by LLETZ under local anaesthetic were included in the study. Patients without follow‐up data were excluded from the study. The excision margin status was correlated with the subsequent cytological and histological outcomes. Results: A series of 967 women with CIN2 and CIN3 underwent LLETZ excision under local anaesthetic. Overall, 42% of women had disease present at the excision margin following LLETZ. Women with CIN3 were more likely than those with CIN2 to have an involved excision margin (P < 0.0001). Cytological recurrence was highest at 12 months (16%) and did not correlate with the CIN grade or excision margin status. Histological recurrence/persistence was also highest at 12 months follow‐up (15%) and this correlated with grade of CIN and margin status (P < 0.0001). Conclusions: Histological recurrence/persistence correlates with grade of CIN and excision margin status. Management of HG‐CIN in an outpatient setting under local anaesthetic is safe, cost effective and yields a favourable long‐term outcome.     

Immunocytochemistry: an indispensable technique in routine cytology

L. Skoog and E. Tani Immunocytochemistry: an indispensable technique in routine cytology Immunocytology is today accepted as an indispensable adjunct to cytomorphology. It has led to a dramatic increase in diagnostic accuracy and also allowed the identification of markers both for prognosis and targeted therapies. Most commercially available antibodies will perform in a reproducible and reliable way provided that the cytological specimen has been prepared and fixed properly. In this review various aspects of immunocytochemistry such as preparation of cytological specimens, fixation and choice of antibodies will be discussed. The specificity of the most commonly used antibodies is summarized and staining panels for various tumours are suggested. In addition, the use of markers for targeted therapy and theranostics is discussed, as well as a brief section on the identification of infectious agents.     

大果白刺雄性不育系的细胞学研究

利用石蜡切片法对大果白刺花药进行细胞学研究,探讨雄性不育系发生败育的时期和方式以及雄性败育与药壁组织间的关系.结果表明:不育系小孢子母细胞形成前期,花药各部分结构发育正常.随着绒毡层的异常解体,多种异常现象相继出现,包括小孢子母细胞液泡化,中层、药室内壁、药隔细胞液泡化,细胞畸形,药壁细胞非正常解体等.退化后整个花药萎缩干瘪,不能开裂,无花粉.因此,大果白刺雄性不育系的绒毡层生理异常并提前退化是导致雄性不育的主要原因.     

人乳头状瘤病毒DNA检测在宫颈病变筛查中的应用研究

目的：探讨高危型人乳头状瘤病毒（HR-HPV）DNA检测方法在宫颈病变筛查中的应用意义。方法：580例妇女同时进行薄层液基细胞学（TCT）、第2代杂交捕获法（HC Ⅱ）和阴道镜下宫颈组织活检,并以病理组织学检查结果作为确诊标准进行对比分析。结果：①580例受检者中病理诊断为炎症207例（35.69%）,CIN Ⅰ 224例（38.62%）,CIN Ⅱ 96例（16.55%）,CIN Ⅲ 38例（6.55%）,浸润癌15例（2.58%）;②TCT检测异常者中炎症52例（25.12%）,CIN Ⅰ 177例（79.02%）,CIN Ⅱ 85例（88.54%）,CIN Ⅲ36例（94.74%）,浸润癌15例（100%）,其中CIN Ⅱ和CIN Ⅲ组间差异无统计学意义（P〉0.05）,但显著高于炎症组和CIN Ⅰ组,低于湿润癌组（P〈0.01或0.05）;③HPV DNA检测阳性者中炎症66例（31.88%）,CIN Ⅰ 152例（67.86%）,CIN Ⅱ 83例（86.46%）,CINIII 35例（92.11%）,浸润癌组15例（100%）,除CIN Ⅱ和CIN Ⅲ组间差异无显著性外（P〉0.05）,其余各组间差异均有统计学意义（P〈0.05或0.01）,且HPV-DNA检测阳性组CIN和浸润癌发病率明显高于阴性组（P〈0.01）;④30岁以下高危险型HPV感染率（65.53%）显著高于30岁以上34.47%感染率（P〈0.01）;⑤联合应用TCT、HPV-DNA检测诊断宫颈癌及癌前病变的敏感度和特异度分别为96.14%和69.28%,高于TCT或HPV-DNA的单独检测。结论：宫颈高危险型HPV感染是CIN及宫颈癌的主要发病因素,并与病变严重程度密切相关,而HPV-DNA和TCT联合应用可提高宫颈癌及癌前病变的检出率。     

The role of liquid-based cytology associated with curettage in the investigation of endometrial lesions from postmenopausal women

OBJECTIVE: This study investigates the role of liquid-based cytology with ThinPrep technique, in the detection of endometrial lesions, using direct endometrial sampling from postmenopausal women with the Endogyn endometrial device. METHODS: It was performed on 491 postmenopausal women referred to our clinic for abnormal bleeding or other symptoms and/or a thickness of endometrium >5 mm on ultrasound. Endometrial sampling, dilatation and curettage (D&C) and hysterectomy were performed on all patients. For the diagnosis, the WHO classification scheme was used. RESULTS: According to our findings a sensitivity of 98.08%, specificity of 100%, positive predictive value of 100%, negative predictive value of 100% and overall accuracy of 98.98% were observed in both endometrial sampling and in D&C. CONCLUSIONS: Endometrial sampling is complementary to D&C for the diagnosis of endometrial lesions and it is necessary for it to be performed before D&C and/or hysterectomy.     

Optimal molecular profiling of tissue and tissue components

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of any tissue-based biological phenomenon. This article reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genotype. The quality of samples isolated from tissue and used for molecular analysis is often glossed over or omitted from publications, making interpretation and replication of data difficult or impossible. Fortunately, recently developed techniques allow life scientists to better document and control the quality of samples used for a given assay, creating a foundation for improvement in this area. Tissue processing for molecular studies usually involves some or all of the following steps: tissue collection, gross dissection/identification, fixation, processing/embedding, storage/archiving, sectioning, staining, microdissection/annotation, and pure analyte labeling/identification and quantification. We provide a detailed comparison of some current tissue microdissection technologies, and provide detailed example protocols for tissue component handling upstream and downstream from microdissection. We also discuss some of the physical and chemical issues related to optimal tissue processing, and include methods specific to cytology specimens. We encourage each laboratory to use these as a starting point for optimization of their overall process of moving from collected tissue to high quality, appropriately anatomically tagged scientific results. In optimized protocols is a source of inefficiency in current life science research. Improvement in this area will significantly increase life science quality and productivity. The article is divided into introduction, materials, protocols, and notes sections. Because many protocols are covered in each of these sections, information relating to a single protocol is not contiguous. To get the greatest benefit from this article, readers are advised to read through the entire article first, identify protocols appropriate to their laboratory for each step in their workflow, and then reread entries in each section pertaining to each of these single protocols.     

辣椒雄性不育系与可育系小孢子发生的细胞学观察

为了探讨辣椒雄性不育花药败育时期和方式,以辣椒雄性不育系1442A、13733A及其可育系为试材,进行了研究。结果发现：败育现象从造孢细胞时期以后每个阶段都有发生,败育形式有造孢细胞液泡化、畸形、拉长、细胞间隙大;绒毡层细胞径向过度伸长,高度液泡化,且出现多层细胞,严重挤压小孢子母细胞,解体较晚且充塞花粉囊室;薄壁细胞取代了药室内壁、中层、绒毡层和小孢子母细胞的分化;药室内壁、中层层数增加,绒毡层细胞肥大,造孢细胞或花粉母细胞分解解体;由于花粉母细胞胼胝质壁不降解而无法释放出四分体小孢子;染色浅、细胞质被降解成空壳的单核期小孢子因缺乏营养物质而败育。     

五种紫萁属植物的核型分析

研究了5种紫萁属Osmunda植物的核形态学特征, 其间期核属于复杂染色中心型, 有丝分裂前期染色体为中间型, 对有丝分裂前期染色体的动态变化过程和在细胞核内的自然排布情况进行了观察。核型公式为: 粗齿紫萁O. banksiifolia (Presl) Kuhn, 2n=4sm+10st+26t+4T; 紫萁O. japonica Thunb., 2n=2sm+8st+32t(2SAT)+2T;华南紫萁O. vachellii Hook., 2n=4sm+8st+28t+4T; 狭叶紫萁O. angustifolia Ching ex Ching &; Ch. H. Wang, 2n=2sm+4st+34t+4T; 粤紫萁O. mildei C. Chr., 2n=2sm+6st+33t+3T。所有染色体臂比均大于2, 核型类型均为4A, 后3种染色体为首次报道, 且对5种紫萁属植物的核型变异及演化关系进行了讨论, 推测Plenasium亚属3种紫萁属植物中粗齿紫萁核型类型最原始, 狭叶紫萁最进化, 其地理分布与核型不对称性有关联, 粤紫萁可能为一自然种间杂种。     

Diagnosis of deep‐seated lymphomas by endoscopic ultrasound‐guided fine needle aspiration combined with flow cytometry

A. Stacchini, P. Carucci, D. Pacchioni, G. Accinelli, A. Demurtas, S. Aliberti, M. Bosco, M. Bruno, A. Balbo Mussetto, M. Rizzetto, G. Bussolati and C. De Angelis
Diagnosis of deep‐seated lymphomas by endoscopic ultrasound‐guided fine needle aspiration combined with flow cytometry Objective: Although endoscopic ultrasound combined with fine needle aspiration (EUS‐FNA) is rapidly becoming the preferred diagnostic approach for the sampling and diagnosis of gastrointestinal and mediastinal malignancies, there are limited data as to its use in the diagnosis of lymphoproliferative disorders. Therefore, we carried out a retrospective evaluation of the performance of EUS‐guided FNA combined with flow cytometry (FC) as a tool to improve overall sensitivity and specificity in the diagnosis of lymphoma. Methods: Of 1560 patients having EUS‐guided FNA during the period of the study, a total of 56 patients were evaluated by cytology with FC after EUS‐FNA. There was adequate material to perform FC analysis for all but one case. Results: EUS‐FNA‐FC gave a diagnosis of lymphoma in 11 cases and of reactive lymphadenopathy in 20. A specific histological type was defined by FC alone in eight cases. The remaining cases were diagnosed later by cytology and cell block sections: 13 carcinomas, nine granulomatous lymphadenopathies and one mediastinal extramedullary haematopoiesis. One case was considered only suspicious for lymphoma on cytology and FC but was not confirmed on molecular analysis and one had insufficient material for FC. Conclusions: Our results show that a combination of EUS‐FNA‐FC is a feasible and highly accurate method, which may be used for the diagnosis and subtyping of deep‐seated lymphoma, providing a significant improvement to cytomorphology alone both for diagnosis and treatment planning, as long as immunocytochemistry is available for non‐lymphoma cases.