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Application of Neural Networks to the Classification of Pancreatic Intraductal Proliferative Lesions 神经网络在胰腺导管内增生性病变分类中的应用
K. Okoń, R. Tomaszewska, K. Nowak, J. Stachura
The aim of the study was to test applycability of neural networks to classification of pancreatic intraductal proliferative lesions basing on nuclear features, especially chromatin texture. Material for the study was obtained from patients operated on for pancreatic cancer, chronic pancreatitis and other tumours requiring pancreatic resection. Intraductal lesions were classified as low and high grade as previously described. The image analysis system consisted of a microscope, CCD camera combined with a PC and AnalySIS v. 2.11 software. The following texture characteristics were measured: variance of grey levels, features extracted from the grey levels correlation matrix and mean values, variance and standard deviation of the energy obtained from Laws matrices. Furthermore we used moments derived invariants and basic geometric data such as surface area, the minimum and maximum diameter and shape factor. The sets of data were randomly divided into training and testing groups. The training of the network using the back‐propagation algorithm, and the final classification of data was carried out with a neural network simulator SNNS v. 4.1. We studied the efficacy of networks containing from one to three hidden layers. Using the best network, containing three hidden layers, the rate of correct classification of nuclei was 73%, and the rate of misdiagnosis was 3%; in 24% the network response was ambiguous. The present findings may serve as a starting point in search for methods facilitating early diagnosis of ductal pancreatic carcinoma.
本研究的目的是测试基于核特征,特别是染色质结构的神经网络在胰腺导管内增生性病变分类中的适用性。该研究的材料来自于胰腺癌、慢性胰腺炎和其他需要胰腺切除术的肿瘤手术患者。如前所述,导管内病变分为低级别和高级别。图像分析系统由一台显微镜、CCD相机和一台PC机以及analysis v. 2.11软件组成。测量纹理特征:灰度方差、灰度相关矩阵提取的特征和均值、Laws矩阵提取的能量方差和标准差。在此基础上,利用矩导不变量和基本几何数据,如表面积、最小和最大直径以及形状因子。这些数据被随机分为训练组和测试组。使用反向传播算法对网络进行训练,并使用神经网络模拟器SNNS v. 4.1对数据进行最终分类。我们研究了包含一到三个隐藏层的网络的有效性。使用包含3个隐藏层的最佳网络,核分类正确率为73%,误诊率为3%;24%的网络反应是模糊的。本研究结果可作为寻找早期诊断导管性胰腺癌方法的起点。
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引用次数: 10
Cytologic and DNA-Cytometric Early Diagnosis of Oral Cancer 口腔癌的细胞学和dna细胞学早期诊断
T. Remmerbach, H. Weidenbach, N. Pomjanski, K. Knops, S. Mathes, A. Hemprich, A. Böcking
Objective. The aim of this prospective study was to report on the diagnostic accuracy of conventional oral exfoliative cytology taken from white‐spotted, ulcerated or other suspicious oral lesions in our clinic. In addition we checked DNA‐image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA‐aneuploidy is a sensitive and specific marker for the early identification of tumor cells in oral brushings. Study design. 251 cytological diagnoses obtained from exfoliative smears of 181 patients from macroscopically suspicious lesions of the oral mucosa and from clinically seemingly benign oral lesions which were exisiced for establishing histological diagnoses were compared with histological and/or clinical follow‐ups of the respective patients. Additionally nuclear DNA‐contents were measured after Feulgen restaining using a TV image analysis system. Results. Sensitivity of our cytological diagnosis on oral smears for the detection of cancer cells was 94.6%, specificity 99.5%, positive predictive value 98.1% and negative predictive value 98.5%. DNA‐aneuploidy was assumed if abnormal DNA‐stemlines or cells with DNA‐content greater 9c were observed. On this basis the prevalence of DNA‐aneuploidy in smears of oral squamous cell carcinomas in situ or invasive carcinomas was 96.4%. Sensitivity of DNA‐aneuploidy in oral smears for the detection of cancer cells was 96.4%, specificity 100%, positive predictive value 100% and negative 99.0%. The combination of both techniques increased the sensivity to 98.2%, specificity to 100%, positive predictive value to 100% and negative to 99.5%. Conclusions. Brush cytology of all visible oral lesions, if they are clinically considered as suspicious for cancer, are an easily practicable, cheap, non‐invasive, painless, safe and accurate screening method for detection of oral precancerous lesions, carcinoma in situ or invasive squamous cell carcinoma in all stages. We conclude that DNA‐image cytometry is a very sensitive, highly specific and objective adjuvant tool for the early identification of neoplastic epithelial cells in oral smears.
目标。本前瞻性研究的目的是报告在我们的诊所中从白斑、溃疡或其他可疑口腔病变中提取的传统口腔剥脱细胞学诊断的准确性。此外,我们检查了DNA图像细胞术作为辅助诊断工具。我们的假设是DNA -非整倍体是一个敏感和特异性的标记物,可以早期识别口腔刷牙中的肿瘤细胞。研究设计。对181例口腔黏膜宏观可疑病变患者和临床表现为良性的口腔病变患者的剥脱涂片进行细胞学诊断,并与各自患者的组织学和/或临床随访进行比较。此外,使用电视图像分析系统测量Feulgen保留后的核DNA含量。结果。口腔涂片细胞学诊断检测癌细胞的敏感性为94.6%,特异性为99.5%,阳性预测值为98.1%,阴性预测值为98.5%。如果观察到异常的DNA干系或DNA含量大于9c的细胞,则假定DNA非整倍体。在此基础上,口腔鳞状细胞原位癌或浸润性癌涂片中DNA -非整倍体的患病率为96.4%。DNA -非整倍体口腔涂片检测癌细胞的敏感性为96.4%,特异性为100%,阳性预测值为100%,阴性预测值为99.0%。两种技术联合使用的敏感性为98.2%,特异性为100%,阳性预测值为100%,阴性预测值为99.5%。结论。对所有可见的口腔病变进行刷细胞学检查,如果临床认为可能是癌症,是一种简便、廉价、无创、无痛、安全、准确的筛查方法,可用于检测口腔癌前病变、原位癌或侵袭性鳞状细胞癌的各个阶段。我们得出结论,DNA图像细胞术是一种非常敏感、高度特异性和客观的辅助工具,可用于口腔涂片中肿瘤上皮细胞的早期鉴定。
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引用次数: 122
High Resolution Comparative Genomic Hybridization Detects 7–8 Megabasepair Deletion in PCR Amplified DNA1 高分辨率比较基因组杂交检测PCR扩增的DNA1中7-8兆对缺失
J. Larsen, A. M. Ottesen, M. Kirchhoff, C. Lundsteen, J. Larsen
We investigated if any change in spatial resolution of comparative genomic hybridization analysis could be detected when using DNA amplified by degenerate oligonucleotide primed PCR (DOP‐PCR) as opposed to the use of unamplified DNA. Five DNA samples from B‐cell leukemias with small 11q deletions were amplified by DOP‐PCR and analysed by means of high resolution comparative genomic hybridization (HR‐CGH) for the evaluation of aberration size detection limit. By means of HR‐CGH, we found the detection limit of DOP‐PCR CGH for deletions to be between 3 Mbp and 7–8 Mbp.
我们研究了与使用未扩增的DNA相比,使用退化寡核苷酸引物PCR (DOP‐PCR)扩增的DNA是否可以检测到比较基因组杂交分析的空间分辨率变化。用DOP - PCR扩增了5例11q缺失的B细胞白血病DNA样本,并用高分辨率比较基因组杂交(HR - CGH)进行分析,以评估畸变大小的检测限。通过HR - CGH,我们发现DOP - PCR CGH对缺失的检出限在3 Mbp到7-8 Mbp之间。
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引用次数: 7
DNA Ploidy and S-Phase Fraction in Carcinoma of the Gallbladder Related to Histopathology, Number of Gallstones and Survival 1 胆囊癌DNA倍性和s期分数与组织病理学、胆结石数量和生存率的关系
U. Gustafsson, C. Einarsson, L. Eriksson, V. Gadaleanu, S. Sahlin, B. Tribukait
Gallstones are a risk factor for the development of gallbladder cancer. We studied DNA ploidy and cell cycle composition by flow cytometry in archival specimens from 52 gall bladder carcinomas in relation to histopathological grade, tumour stage, gallstone number and survival. 69% of the gallbladder carcinomas showed aneuploidy. All tumours with single stones (N=11) were aneuploid while only 61% of tumours with multiple stones (N=41) were aneuploid (p=0.002). DNA aneuploidy was related to increase in T‐category (p=0.01), grade (p=0.02), and nuclear pleomorphism (p=0.0005). The distribution of DNA ploidy shifted from tetraploid in low stage towards triploid positions in high stage tumours (p=0.02) combined with higher S‐phase values in triploid tumours (p=0.05). S‐phase fraction increased during development from normal tissue to dysplasia, cancer in situ and cancer in diploid cases (p=0.0002), and further at the change from diploid to aneuploid (p=0.004). At a median cancer specific survival time of four months patients with diploid tumours had a better survival than those with aneuploid tumours (p=0.02). In multivariate analysis of the tumour characteristic, only T‐category and tumour grade were independent prognostic factors. The shift from diploid to aneuploid and the further shift of ploidy within aneuploid tumours are in agreement with the concept of a clonal development of gallbladder cancer. These changes are combined with a stepwise increase in the fraction of S‐phase cells. Low frequency of symptoms in single stone patients may be the reason for detection of malignancy at a late stage of tumour development.
胆结石是患胆囊癌的危险因素之一。我们用流式细胞术研究了52例胆囊癌档案标本的DNA倍体和细胞周期组成与组织病理分级、肿瘤分期、胆结石数量和生存期的关系。69%的胆囊癌呈非整倍体。所有合并单个结石的肿瘤(N=11)为非整倍体,而合并多个结石的肿瘤(N=41)中只有61%为非整倍体(p=0.002)。DNA非整倍性与T型(p=0.01)、等级(p=0.02)和核多形性(p=0.0005)的增加有关。DNA倍体的分布从低期的四倍体向高期肿瘤的三倍体转移(p=0.02),三倍体肿瘤的S期值更高(p=0.05)。在正常组织向发育不良、原位癌和二倍体癌的发育过程中,S期分数增加(p=0.0002),在二倍体向非整倍体的发育过程中,S期分数增加(p=0.004)。在4个月的中位癌症特异性生存时间中,二倍体肿瘤患者比非整倍体肿瘤患者有更好的生存(p=0.02)。在肿瘤特征的多变量分析中,只有T型和肿瘤分级是独立的预后因素。从二倍体到非整倍体的转变以及在非整倍体肿瘤中倍性的进一步转变与胆囊癌克隆发展的概念一致。这些变化与S相细胞比例的逐步增加相结合。单一结石患者的低频率症状可能是在肿瘤发展晚期才发现恶性肿瘤的原因。
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引用次数: 4
Fibroblast Growth Factor Receptor 3 (FGFR3)–Analyses of the S249C Mutation and Protein Expression in Primary Cervical Carcinomas 成纤维细胞生长因子受体3 (FGFR3) -原发性宫颈癌中S249C突变和蛋白表达分析
Haiyan Dai, R. Holm, G. Kristensen, V. Abeler, A. Børresen-Dale, Å. Helland
Fibroblast growth factor receptor 3 (FGFR3) seems to play an inhibitory role in bone development, as activating mutations in the gene underlie disorders such as achondroplasia and thanatophoric dysplasia. Findings from multiple myeloma (MM) indicate that FGFR3 also can act as an oncogene, and mutation of codon 249 in the fibroblast growth factor receptor 3 (FGFR3) gene was recently detected in 3/12 primary cervical carcinomas. We have analysed 91 cervical carcinomas for this specific S249C mutation using amplification created restriction site methodology (ACRS), and detected no mutations. Immunohistochemistry was performed on 73 of the tumours. Reduced protein staining was seen in 43 (58.8%) samples. Six of the tumours (8.2%) revealed increased protein staining compared with normal cervical tissue. These patients had a better prognosis than those with reduced or normal levels, although not statistically significant. This report weakens the hypothesis of FGFR3 as an oncogene of importance in cervical carcinomas.
成纤维细胞生长因子受体3 (FGFR3)似乎在骨骼发育中起抑制作用,因为该基因的激活突变是软骨发育不全和嗜盐性发育不良等疾病的基础。多发性骨髓瘤(MM)的研究结果表明,FGFR3也可以作为致癌基因,最近在3/12的原发性宫颈癌中检测到成纤维细胞生长因子受体3 (FGFR3)基因密码子249的突变。我们使用扩增限制性内切位点方法(ACRS)分析了91例宫颈癌中这种特定的S249C突变,未检测到突变。73例肿瘤行免疫组化处理。43例(58.8%)蛋白染色降低。与正常宫颈组织相比,6例肿瘤(8.2%)显示蛋白染色增加。这些患者的预后比低水平或正常水平的患者好,尽管没有统计学意义。该报告削弱了FGFR3作为宫颈癌重要致癌基因的假设。
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引用次数: 10
In Situ Reverse Transcriptase-Nested Polymerase Chain Reaction to Identify Intracellular Nucleic Acids without the Necessity of Dnase Pretreatment and Hybridisation 原位逆转录-巢式聚合酶链反应鉴定细胞内核酸无需dna酶预处理和杂交
M. Menschikowski, M. Vogel, R. Eckey, G. Dinnebier, W. Jaross
In the present study a protocol of in situ reverse transcriptase‐nested polymerase chain reaction (in situ RT‐nested PCR) was examined based on the following modifications. (i) To exclude false positive signals caused by “DNA repair mechanisms” and “endogenous priming”, a two‐step PCR was applied after reverse transcription. The first step was performed in the presence of extrinsic primers and unlabeled nucleotides with a maximum of PCR cycles possible without destroying the cell morphology. The second step consisted of only one annealing/elongation reaction, the target sequence marked by addition of digoxigenin‐labeled nucleotides and intrinsic primers. (ii) In order to prevent amplifications of genomic DNA nested primer pairs were applied crossing intron sequences. (iii) To minimize the diffusion of PCR products in cells, the extrinsic primers were extended with complementary 5′‐tails. This approach results in the generation of high molecular weight concatamers during PCR cycles. By applying this protocol, immunostainings specific for phospholipase A2 of type IIA mRNA were exclusively detectable in the cytoplasm of HepG2 hepatoma cells, which were used as a model system, whereas the nuclei were unstained. Multiple control experiments yielded completely negative results. These data suggest that the in situ RT‐nested PCR, which in comparison to the method of in situ RT‐PCR‐in situ‐hybridisation is simpler and less time‐consuming, can be used as an alternative approach to identify intracellular nucleic acids.
在本研究中,基于以下修改,对原位逆转录酶巢式聚合酶链反应(原位RT巢式PCR)方案进行了检验。(i)为了排除由“DNA修复机制”和“内源性引物”引起的假阳性信号,逆转录后采用两步PCR。第一步是在外部引物和未标记的核苷酸存在的情况下进行的,在不破坏细胞形态的情况下可能进行最多的PCR循环。第二步仅包括一个退火/延伸反应,目标序列通过添加地高辛标记的核苷酸和内在引物来标记。(ii)为了防止基因组DNA的扩增,在内含子序列间应用嵌套引物对。(iii)为了尽量减少PCR产物在细胞中的扩散,外源引物用互补的5 '‐尾巴延伸。这种方法导致在PCR周期中产生高分子量的连接体。通过应用该方案,在HepG2肝癌细胞的细胞质中可以检测到IIA型mRNA的磷脂酶A2特异性免疫染色,而细胞核未染色。多次对照实验得出了完全否定的结果。这些数据表明,与原位RT - PCR -原位杂交相比,原位RT -巢式PCR更简单,耗时更少,可以作为鉴定细胞内核酸的替代方法。
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引用次数: 1
7th Congress of the European Society for Analytical Cellular Pathology, Caen, France, 1–5 April 2001 第七届欧洲分析细胞病理学学会大会,法国卡昂,2001年4月1-5日
M. Ormerod
One of the attractions of a career in science is the opportunity to make friends with people across the globe. A parallel advantage is the opportunity to visit attractive cities in a variety of countries. Sometimes, the science is an added bonus! Caen, in Normandy, combined all three – good science , good company and excellent sightseeing. Although 70% of the city was unfortunately destroyed during the Second World War, much was preserved or has now been restored. This is the city of William the Conqueror who was buried here in the abbey he founded, L'Abbaye aux Hommes. The remains of his castle are opposite the University and the venue for the 7th ESACP Conference. The bottle of cider on the lunch timetable reminded me that I was truly in the heart of Normandy. However , the organisers ensured that we had time to appreciate some of the other delights of the region. We were welcomed to Caen by the Mayor at a reception in the second large abbey, L'Abbaye aux Dames, founded by William's wife, Mathilde, in 1067. On the penultimate day, a coach tour of the Fleurie Coast visited Honfleur and Deauville. We finished the tour in Beuvron-en-Auge, a typical Norman village in the " Pays d'Auge " , at a converted manor, 'Haras de Sens'. Alfred Böck-ing gave the Distinguished Ploem Lecture entitled 'To-wards a non-invasive, objective single cell cancer diag-nosis'. Haras de Sens is famous for its horses trained for dressage. An exhibition of these skills accompanied by a local brass band was followed by an excellent meal, introducing us to some of the regional cuisine. There was also some first class science. I will not give you a blow by blow account of the lectures – you can read the abstracts in an earlier volume of this journal. The highlight of the plenary sessions was probably Oli Kallioniemi's lecture on 'Tissue Microarray (" tissue chip ") Technology for high-throughput molecular profiling of cancer'. In the parallel sessions, it was heartening to hear so many good papers from younger scientists. The Organ-isers had underlined the importance of youth by devoting a large part of the available funds to the sponsorship of students. The poster that did not win a prize but drew the most attention was an evaluation of the biological safety of condoms by Pretorius et al. tested on in vitro cultures. While spermicides had …
从事科学事业的吸引力之一是有机会与世界各地的人交朋友。另一个优势是有机会参观不同国家的迷人城市。有时候,科学是一个额外的奖励!位于诺曼底的卡昂集三者于一身:良好的科学、良好的伙伴和绝佳的观光。虽然70%的城市不幸在第二次世界大战期间被摧毁,但大部分都被保存下来或现在已经修复。这是征服者威廉的城市,他被埋葬在这里,他建立的修道院,L'Abbaye aux Hommes。他的城堡遗迹就在大学对面,也是第七届ESACP会议的举办地。午餐时间表上的那瓶苹果酒提醒我,我真的身处诺曼底的中心。然而,组织者确保我们有时间欣赏该地区的其他一些乐趣。我们来到卡昂,市长在第二大修道院——由威廉的妻子玛蒂尔德(Mathilde)于1067年建立的圣母院(L’abbaye aux Dames)的招待会上欢迎我们。在倒数第二天,Fleurie海岸的长途旅行参观了Honfleur和Deauville。我们在Beuvron-en-Auge结束了旅程,这是一个典型的诺曼村庄,位于“Pays d' auge”,在一个改造的庄园“Haras de Sens”。Alfred Böck-ing发表了题为“迈向非侵入性、客观的单细胞癌诊断”的杰出Ploem演讲。哈拉斯德森斯以其训练有素的马匹而闻名。在当地铜管乐队的伴奏下,他们展示了这些技巧,之后是一顿丰盛的晚餐,向我们介绍了一些当地的美食。还有一些一流的科学。我就不逐字逐句地给你讲讲座了,你可以在这本杂志的前一卷中读到摘要。全体会议的亮点可能是Oli Kallioniemi关于“组织微阵列(“组织芯片”)技术用于高通量癌症分子分析”的演讲。在平行会议上,听到这么多年轻科学家的好论文令人振奋。主办方强调了青年的重要性,将大部分可用资金用于资助学生。这张海报虽然没有获奖,但却引起了最多的关注,它是Pretorius等人在体外培养中对避孕套的生物安全性进行的评估。而杀精剂……
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引用次数: 1
Amplification at 9p in Cervical Carcinoma by Comparative Genomic Hybridization 比较基因组杂交法扩增宫颈癌9p位点
K. Jee, Young Tak Kim, Kyu Rae Kim, Y. Aalto, S. Knuutila
DNA copy number changes were studied by comparative genomic hybridization on 10 tumor specimens of squamous cell carcinoma of cervix obtained from Korean patients. DNA was extracted from paraffin‐embedded sections after removal of non‐malignant cells by microdissection technique. Copy number changes were found in 8/10 tumors. The most frequent changes were chromosome 19 gains (n=6) and losses on chromosomes 4 (n=4), 5 (n=3), and 3p (n=3). A novel finding was amplification in chromosome arm 9p21‐pter in 2 cases. Gains in 1, 3q, 5p, 6p, 8q, 16p, 17, and 20q and losses at 2q, 6q, 8p, 9q, 10p, 11, 13, 16q, and 18q were observed in at least one of the cases.
采用比较基因组杂交技术,对10例韩国人宫颈鳞状细胞癌肿瘤标本的DNA拷贝数变化进行了研究。用显微解剖技术去除非恶性细胞后,从石蜡包埋切片中提取DNA。8/10的肿瘤出现拷贝数改变。最常见的变化是19号染色体的增加(n=6)和4号染色体(n=4)、5号染色体(n=3)和3p号染色体(n=3)的减少。一个新的发现是染色体臂9p21 - pter在2例中扩增。至少有一种情况在第1、3、5、6、8、16、17和20季度出现收益,在第2、6、8、9、10、11、13、16和18季度出现亏损。
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引用次数: 9
Topical Mitomycin C and Radiation Induce Conjunctival DNA-Polyploidy 局部丝裂霉素C和辐射诱导结膜dna多倍体
O. Cartsburg, C. Kallen, J. Hillenkamp, R. Sundmacher, N. Pomjanski, A. Böcking
Introduction: Atypical cell changes often occur following treatment of premalignant or malignant conjunctival neoplasias with topical mitomycin C (MMC) and/or radiation. These reactive, non‐neoplastic alterations of the conjunctival epithelium can be a differential diagnostic problem. Our aim was to investigate changes in the nuclear DNA‐distribution of conjunctival epithelial cells after MMC‐ and radiation therapy by DNA‐image‐cytometry. Methods: Conjunctival brush smears were obtained from 13 patients (13 eyes) with squamous cell carcinomas and six patients (6 eyes) with conjunctival malignant melanomas in situ before, during and after treatment. The patients were treated with MMC‐drops (0.02% or 0.04%) alone (n=12), with radiation therapy (n=3) or both (n=4). At first, the obtained brush smears were evaluated by cytology. Secondly, after Feulgen restaining, the DNA content of reactively changed cells was determined using the AutoCyte‐QUIC‐DNA® workstation. Results: We observed euploid DNA‐polyploidy and cytomorphological changes in all patients (19/19). We considered these alterations as reactive to treatment. Four patients showed their greatest DNA‐stemline at 4c and 15 patients at 8c. This effect was observed during and following MMC‐drops and/or radiation and remained stable in 94% of all patients after a mean follow‐up of 22.5 months (SD 15.4). In five cases image cytometry additionally demonstrated DNA‐stemline aneuploidy as an evidence of tumor recurrence. Conclusion: Measurements of DNA‐content revealed euploid polyploidisation of morphological suspicious but benign squamous cells which is the biologic correlate of well known secondary morphologic changes following topical chemotherapy and/or radiation. DNA‐image‐cytometry is a useful tool in the differention of euploid polyploidization as a sign of reactive cell changes following treatment and tumor recurrences.
非典型细胞改变常发生在外用丝裂霉素C (MMC)和/或放射治疗癌前或恶性结膜瘤后。结膜上皮的这些反应性、非肿瘤性改变可能是鉴别诊断的问题。我们的目的是通过DNA图像细胞术研究MMC和放射治疗后结膜上皮细胞核DNA分布的变化。方法:对13例(13眼)鳞状细胞癌患者和6例(6眼)原位结膜恶性黑色素瘤患者在治疗前、治疗中及治疗后进行结膜刷涂片。患者分别接受MMC滴剂(0.02%或0.04%)单独治疗(n=12)、放射治疗(n=3)或两者同时治疗(n=4)。首先,对获得的毛刷涂片进行细胞学评价。其次,在Feulgen抑制后,使用AutoCyte - QUIC - DNA®工作站测定反应改变细胞的DNA含量。结果:我们在所有患者中观察到整倍体DNA -多倍体和细胞形态学变化(19/19)。我们认为这些改变是对治疗的反应。4例患者在4c和15例患者在8c时显示出最大的DNA - stemline。在MMC滴注和/或放疗期间和之后观察到这种效应,在平均随访22.5个月后,94%的患者保持稳定(标准差15.4)。在5个病例中,图像细胞术还显示DNA -茎系非整倍体是肿瘤复发的证据。结论:DNA含量测量显示形态学可疑但良性的鳞状细胞整倍体多倍体化,这是众所周知的局部化疗和/或放疗后继发形态学变化的生物学相关性。DNA -图像-细胞术在整倍体多倍体分化中是一种有用的工具,作为治疗和肿瘤复发后反应性细胞变化的标志。
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引用次数: 13
Comparative Intracellular Cytokine Production by In Vitro Stimulated T Lymphocytes from Human Umbilical Cord Blood (HUCB) and Adult Peripheral Blood (APB) 体外刺激人脐带血(HUCB)和成人外周血(APB) T淋巴细胞胞内细胞因子生成的比较
J. G. García Vela, I. Delgado, R. Bornstein, B. Alvarez, M. C. Auray, I. Martín, F. Oña, F. Gilsanz, J. Antônio, García Vela
To date over 400 HUCB transplants have been reported from different centers. It has been suggested that there is a reduced graft‐versus‐host‐disease (GVHD) with HUCB compared to bone marrow transplantation. Since cytokine production by a cell is an indication of the cells function it is important to determinate the differences between APB and HUCB with respect to production of these soluble factors. Our aim was to analyse the intracellular cytokine production by HUCB and APB T lymphocytes with and emphasize on their possible role in GVHD. Heparinized HUCB samples from 8 normal full‐term deliveries and 10 normal blood donors were stimulated 4 hours at 37∘C and 5% CO2 with phorbol 12‐myristate 13‐acetate (PMA) and Ionomycin in the presence of brefeldine. Afterwards cells were stained with CD3, CD4 or CD8 in different combinations. Finally, after cell permeabilization, cells were stained with Il‐2, Il‐4 or IFN‐γ. Data acquisition was performed on a FACScan flow cytometer. Compared to APB, HUCB T lymphocytes produced less Il‐2, Il‐4 and IFN‐γ. In HUCB, Il‐2, Il‐4 and IFN‐γ were produced predominantly by CD4+ T cells. In APB, Il‐2 and Il‐4 were also produced predominantly by CD4+ cells compared with CD8+ T lymphocytes, however, IFN‐γ was produced by both CD4+ and CD8+ T cells. These results indicate that there are clear differences in the cytokine profile between T cells in APB and HUCB.
迄今为止,不同中心报道了400多例hub移植。有研究表明,与骨髓移植相比,hub的移植物抗宿主病(GVHD)减少。由于细胞产生的细胞因子是细胞功能的一个指标,因此确定APB和hub在产生这些可溶性因子方面的差异是很重要的。我们的目的是分析hub和APB T淋巴细胞细胞内细胞因子的产生,并强调它们在GVHD中的可能作用。在brefeldine存在的情况下,用phorbol 12‐肉豆酸酯13‐乙酸酯(PMA)和Ionomycin在37°C和5% CO2下刺激8例正常足月分娩和10例正常献血者的肝素化HUCB样本4小时。然后用不同组合的CD3、CD4或CD8染色细胞。最后,在细胞渗透后,用Il - 2、Il - 4或IFN - γ对细胞进行染色。数据采集在FACScan流式细胞仪上进行。与APB相比,HUCB T淋巴细胞产生较少的Il - 2、Il - 4和IFN - γ。在hub中,Il - 2、Il - 4和IFN - γ主要由CD4+ T细胞产生。在APB中,与CD8+ T细胞相比,Il‐2和Il‐4也主要由CD4+细胞产生,然而,IFN‐γ由CD4+和CD8+ T细胞产生。这些结果表明,APB和HUCB中T细胞的细胞因子谱存在明显差异。
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引用次数: 12
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Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology
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