目的 比较Victoria blue法(A组)和Verhoeff法(B组)弹力纤维染色在染色过程、染色时间、染色效果方面对肺癌胸膜侵犯和结直肠癌浆膜或静脉侵犯判读的差异.方法 A组、B组均分别纳入肺癌怀疑胸膜侵犯标本50例和结直肠癌怀疑浆膜或静脉侵犯标本20例,进行弹力纤维染色.对比2组染色过程、染色时间、染色效果;根据染色稳定性、清晰度、对比度等综合评价胸膜侵犯和浆膜或静脉侵犯情况,并对染色质量及判读准确性进行评分.结果 A组比B组操作简单,染色质量稳定,肿瘤细胞与脏层胸膜、浆膜或静脉的关系显示更清晰.A组判读评分高于B组,差异有统计学意义(P＜0.05).结论 Victoria blue法染色有助于准确评估肺癌的胸膜侵犯和结直肠癌浆膜或静脉的侵犯,提高判读准确性.

Objective:To study the effect of MYD88 L265P mutation on the expression of PD-L1 in tumor cells and tumor microenvironment in diffuse large B-cell lymphoma (DLBCL), and to provide theoretical basis for immunotherapy for patients.Methods:Multiplex ligation-dependent probe amplification (MLPA) was used to detect the frequency of MYD88 L265P mutation in 72 cases of DLBCL diagnosed by pathologists in Cancer Hospital of Chinese Academy of Medical Sciences from August 2008 to May 2010. Expression of PD-L1 in tumor cells and tumor microenvironment in all samples was evaluated using PD-L1 (22C3) and PD-L1 (SP142) with Ventana automatic immunohistochemical (IHC) platform. The relationship between MYD88 L265P mutation and the expression of PD-L1 in DLBCL tumor cells and tumor microenvironment was assessed.Results:Of the 72 cases of DLBCL, MYD88 L265P mutation was detected in 15 (20.8%) cases. Nine cases with JAK2 amplification were excluded, and the remaining 63 cases of DLBCL were divided into MYD88 L265P mutant group ( n=14) and MYD88 L265P wild-type group ( n=49). IHC results showed that among the 14 cases of MYD88 L265P mutant groups, PD-L1 (22C3) was positive in 7 cases (7/14) of tumor cells and PD-L1 (SP142) was positive in 4 cases (4/14) of tumor microenvironment. Among the 49 cases of MYD88 L265P wild-type group, 9 cases (18.4%) were positive for PD-L1 (22C3) in tumor cells, and 38 cases (77.6%) were positive for PD-L1(SP142) in tumor microenvironment. In addition, among the 16 cases with PD-L1(22C3) expression in tumor cells, only 2 of the 7 cases with MYD88 L265P mutation were positive for PD-L1 (SP142) in tumor microenvironment. All 9 cases with wild-type MYD88 L265P were positive for PD-L1 (SP142) in tumor microenvironment. Statistical analysis showed that the expression level of PD-L1 (22C3) in tumor cells in the MYD88 L265P mutant group was significantly higher than that in the MYD88 L265P wild-type group ( P=0.017). The expression level of PD-L1 (SP142) in tumor microenvironment in the MYD88 L265P mutant group was significantly lower than that in the MYD88 L265P wild-type group ( P=0.001). Conclusions:MYD88 L265P mutation may play an important role in the regulation of PD-L1 expression in DLBCL tumor cells and tumor microenvironment. Further studies will provide a theoretical basis for immunotherapy of DLBCL patients with MYD88 L265P mutation.

Objective To explore the application value of treatment-related markers PD-L1, PD-L2, CD30, CD23, BCL-2, BCL-6, MUM1 and GATA3 in the diagnosis and prognostic evaluation of primary mediastinal B-cell lymphoma(PMBL). Methods A retrospective study was conducted on 34 patients diagnosed with PMBL, and 31 patients with DLBCL-NOS which was not primary in the mediastinum were taken as control group. The expressions of 8 proteins were detected by IHC staining. Results The median percentages of tumor cells with PD-L1, PD-L2 and CD30 expression in PMBL group were 70% (30%, 90%), 25% (0, 70%) and 17.5% (0, 60%) respectively, which were significantly higher than those in the DLBCL-NOS group (P < 0.05). The positive rates of CD30 and CD23 in PMBL group were 61.76% (21/34) and 76.47% (26/34) respectively, significantly different with those in the DLBCL-NOS group (P=0.000). The survival curve of PMBL patients with CD30 or BCL-6 expression showed a trend of poor prognosis, despite the P value was > 0.05. Conclusion The high expression levels of PD-L1, PD-L2 and CD30 in PMBL are helpful to accurately identify more patients who may respond to immune or targeted therapy. Immunohistochemical staining of PD-L1, PD-L2, CD30 and CD23 is helpful for the differential diagnosis of PMBL and DLBCL-NOS. As candidate prognostic indicators of PMBL, CD30 and BCL-6 should be further studied in a larger number of samples.

目的 研究结直肠癌人工智能病理诊断模型构建过程中,病理医师对数字切片癌组织的人工标注在不同扫描仪构建的全切片图像(WSI)中准确迁移的方法.方法 在本研究中,我们提出了一种基于图像配准的标注迁移方法,在来自不同扫描仪的WSI之间建立仿射映射.通过多分辨率最小化两个WSI缩略图之间的互信息来估计最佳仿射映射参数,以避免和改变扫描仪特定特性的影响,减少计算时间.我们使用了181张结直肠癌病理切片,使用两个品牌的扫描仪获得相应的WSI,对上述标注迁移方法进行测试.结果 181张HE切片的扫描结果表明,同一张切片由不同扫描仪构建的WSI在颜色、位置、大小等属性上都有不同的表现.使用我们提出的标注迁移方法,其中179张图像的人工标注成功地在不同扫描仪构建的WSI中迁移,其中125对使用单个CPU核心的计算时间不到1分钟.结论 我们提出了一种快速、准确的全自动的标注迁移方法,用于在不同扫描仪构建的WSI之间传递人工标注.在准备深度学习训练数据过程中,既可以避免病理医师对新图像的重新标注,也可以避免病理医师之间在标注上的差异.

在日常工作中,我科批量出现了淋巴结组织染色模糊不清的情况,这些问题切片皆由罗氏SYMPHONY全自动染色机染色处理的,并多为一个包埋盒中放置多枚淋巴结的组织块.对于造成该结果的原因,参照相关文献并结合实验室的实际情况,对可能存在问题的流程进行排查,具体实验方案如下. 1 材料与方法 1.1 材料收取2019年6月中国医学科学院肿瘤医院病理科接收的外科手术标本30例,其中包括结肠、乳腺、甲状腺标本.所用病理级载玻片(世泰)、SYMPHONY全自动染色机、即用型试剂(罗氏),95％乙醇、100％乙醇、二甲苯、哈瑞苏木精染色液(北京益利精细化学品有限公司),伊红(北京中杉金桥生物技术有限公司),DHG-9140 A鼓风干燥箱(北京陆希科技有限公司).

目的 探讨弥漫性大B细胞淋巴瘤(DLBCL)中MAPK10基因甲基化的临床意义.方法 选取2008年1月至2010年7月间中国医学科学院肿瘤医院病理诊断明确的107例DLBCL患者标本进行回顾性研究.检测MAPK10基因甲基化状态,分析MAPK10基因甲基化与DLBCL各临床病理的关系,对65例经利妥昔单抗治疗的DLBCL患者进行预后分析,评估MAPK10基因甲基化在DLBCL中的预后价值.结果 107例DLBCL中,68例(63.6％)患者发生MAPK10基因甲基化.MAPK10基因甲基化与患者年龄、性别、淋巴瘤国际预后指数、Ann Arbor分期、血清乳酸脱氢酶水平、肿瘤亚型、中枢神经系统及骨髓受侵等均无关.MAPK10基因甲基化的DLBCL患者5年总体生存率和5年无进展生存率与无MAPK10基因甲基化者间的差异无统计学意义(P＞0.05).结论 在DLBCL中,广泛存在MAPK10基因甲基化,可能与DLBCL的发生有关,与DLBCL预后无关.

Objective: To investigate the impact of ultrasonic assisted rapid processing technique combined with the environment friendly reagent (which can be utilized in fixing,dehydrating and clearing) on processing tumor biopsy specimens and the subsequent target detection. Methods: Postoperative tissue samples of 56 cases of breast cancer, colorectal cancer, lung cancer, stomach cancer, liver mass, bladder mass, uterus mass were obtained at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences from February to April, 2017. Three specimens ranging in size from 1 to 3 mm were collected from each sample, and were separated into control group (traditional tissue-processing method); experiment group 1 (3.7% neutral buffered formaldehyde fixation, composite environment friendly reagent and ultrasonic assisted rapid processing) and experimental group 2 (composite environment friendly reagent direct fixation, higher temperature and longer time for tissue processing). Two pathologists blinded to the experimental groups scored totally the nuclear, cytoplasmic, and membrane staining of 43 cases of immunohistochemistry (IHC), four HER2 fluorescence in situ hybridization (FISH), 20 extracted DNA quality and four EGFR gene mutation detection in lung adenocarcinoma; the results were compared with the control group. Results: There was no difference in the IHC staining, HER2 FISH, the DNA quality, and EGFR genetic results between experimental group 1 and control group. For experiment group 2, comparing results of IHC staining, HER2 FISH and the quality of DNA, there was no obvious difference from control group and experiment group 1, but might show an increase in the background of IHC staining. The difference between the treatment temperature and time in the experimental group 2 did not affect the results of the gene mutation detection. Conclusions: Environment freindly reagent and ultrasonic assisted rapid processing equipment could be used for rapid processing and diagnosis for tumor biopsies. Using complex environment-friendly reagents supplement fixation, higher treatment temperature and longer treatment time do not significantly affect the IHC, FISH and molecular detection accuracy.

目的 探讨复合型环保试剂对肿瘤活检组织HE制片中的固定、快速超声组织处理仪设定的工作温度及时间等主要因素的影响.方法 选取56例术后组织标本,模拟术前活检标本进行取材,每例样本取材3块,直径为1～3 mm.3块组织分别归入对照组(传统活检组织处理方法)、实验1组(4％中性甲醛固定及复合型环保试剂配合超声快速组织处理仪方法)和实验2组(复合型环保试剂直接固定及超声快速组织处理仪边缘条件方法),实验1组和实验2组分别设定不同的固定方式、处理时间和温度对组织进行处理.2位病理医师盲法独立观察3组的HE切片并评分,采用t检验方法统计组间差异.结果 实验1组与对照组比较,HE片评分没有差异(P＞0.69,95％CI:-0.33-0.498);实验2组与对照组比较,HE片评分没有差异(P＞0.07,95％CI:-0.83-0.04);实验1组与实验2组比较,实验1组HE片评分显著高于实验2组(P ＜0.03,95％CI:0.046-0.92),实验2组评分均值均小于对照组和实验1组,有显著差异.结论 利用复合型环保试剂配合超声快速组织处理仪可以大大缩短组织处理时间,但不恰当的处理温度和时间及用复合型环保试剂直接固定会对肿瘤活检组织HE染色结果产生影响.

目的:为发挥病理网络与信息系统在病理检查全流程中的应用与跟踪,探索新型激光包埋盒打号机工作原理和应用特征.方法:选取活检和术后标本进行批量包埋盒预打印共2500个,将采用传统碳带包埋盒打号机纳入对照组(打印1250个),采用激光包埋盒打号机纳入观察组(打印1250个).两组均与病理信息系统(LIS)连接,记录单个包埋盒打印时间,计算扫描识别率及平均识别时间.比较两组平均打印时间、平均扫描识别时间以及字迹耐磨程度与扫描识别率的差异.比较两组打印稳定性、使用过程中出现的故障情况.结果:观察组平均打印时间(4.72±0.59)s/个,对照组(20.62±1.39)s/个;观察组平均扫描识别时间(0.88±0.29)s/个,对照组(1.21±0.62)s/个,两组比较有差异;两组总体扫描识别率比较差异有统计学意义(x2=16.44,P<0.05);观察组故障率低于对照组.结论:激光打印技术用于病理技术中能够提高包埋盒打印速度、提高工作效率、包埋盒打印质量稳定、为病理网络与信息系统在病理检查全流程应用与跟踪中提供良好的设备基础.

目的 探讨c-myc蛋白表达与c-myc基因易位的相关性,旨在寻找适用于临床c-myc蛋白的界定值,以有效判断c-myc基因易位状态.方法 采用免疫组化法与荧光原位杂交(fluorescence in situ hybridization,FISH)法检测293例弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中c-myc蛋白表达水平与c-myc基因易位状态,分析两者的相关性.结果 293例DLBCL中,30例(10.2％)发生c-myc基因易位,其中29例c-myc蛋白阳性细胞百分率＞40％,1例c-myc蛋白阳性细胞百分率为40％,而c-myc蛋白阳性细胞百分率＜40％者均未发生c-myc基因易位.随着c-myc蛋白表达水平的升高,c-myc基因易位的发生率也随之升高.结论 c-myc蛋白高表达与c-myc基因易位密切相关.c-myc蛋白可用于DLBCL中c-myc基因易位的筛选,建议对c-myc蛋白阳性细胞百分率＞40％者行FISH检测以进一步明确有无c-myc基因易位.

Objective To identify reliable morphological and immunophenotype characteristics for diagnosis,differential diagnosis and evaluation of cancer risk.Methods A total number of 95 cases of colorectal polyp samples were retrieved and examined by histopathological approaches.And the expression of DNA mismatch repair protein(MLH1,MSH2,MSH6),DNA repair enzyme(MGMT),p53 and BRAF protein were detected by immunohistochemistry.Traditional adenoma containing tubular adenoma,villous adenoma and tubulovillous adenoma were included as a reference group.Results None of the hyperplastic polyps without colorectal carcinoma showed loss of MLH1,but 23.1% of hyperplastic polyps accompanied with colorectal carcinoma lost of MLH1 expression.The absence rate of MSH2,MSH6 and MGMT immunohistochemical reactivity in hyperplastic polyps is significantly higher than that in tradition adenomas.It is only 11.8% of hyperplastic polyps without colorectal carcinoma showed BRAF protein positive,in contrast with 46.2% of hyperpplastic polyps with colorectal carcinoma.And the expression of MLH1,MSH2 and MSH6 is predominantly correlated.The expression of each immunomarker in different group of polyps or adenomas showed no relationship with the age of patient and the lesion site.Conclusions The gene of MLH1,MSH2,MSH6,MGMT and BRAF plays an important role in serrated pathway.And the serrated lesion lost the expression of MLH1 or with BRAF protein positive probably has a high risk of cancerization.

目的测定南京地区收集的15株单纯疱疹病毒2型(HSV-2)对阿昔洛韦和伐昔洛韦的敏感性,初步建立空斑法提取阿昔洛韦耐药株。方法Vero细胞培养法分离扩增出临床毒株,直接免疫荧光法分型,并用MTT法测定15株2型毒株对阿昔洛韦、伐昔洛韦的敏感性。体外阿昔洛韦诱导,空斑纯化HSV耐药株。结果15株临床株对阿昔洛韦和伐昔洛韦的半数病毒抑制质量浓度(IC50)分别为0.0215- 0.2799μg/mL(平均0.0766μg/mL),0.0549-0.4575μg/ml,(平均0.1605μg/mL)。生殖器疱疹初发组与复发组、用药组与未用药组对2种药物的敏感性采用Wilcoxon秩和检验,P值均>0.05,差异无统计学意义。从15株HSV-2临床株中未提取到耐药空斑;从HSV-1Sam实验室标准株提取到一个耐药空斑,MTT法测定阿昔洛韦IC50为13.36μg/mL;从HSV-2 333标准株中提取到9个耐药空斑,扩增后MTT法测定其对阿昔洛韦的IC50为1.04-5.08μg/mL。结论目前在南京地区收集的HSV-2型毒株对阿昔洛韦和伐昔洛韦敏感性较高,尚未发现耐药株。体外药物诱导、空斑纯化能较快提取到HSV耐药株。

目的 探讨CD4+CD25bright调节性T细胞在梅毒患者免疫学变化中的作用.方法利用流式细胞仪和单克隆荧光抗体检测梅毒患者外周血中CD4+CD25bright调节性T细胞在CD3+CD4+辅助性T细胞(Th)中的比例.结果血清固定组梅毒患者外周血中调节性T细胞比例较正常人对照组显著增高(P＜0.01),而一期、二期及潜伏梅毒患者调节性T细胞比例与正常人对照组差异无统计学意义(P＞0.05).各组梅毒患者外周血调节性T细胞比例与血清RPR滴度无相关性(P＞0.05).结论梅毒患者外周血中CD4+CD25bright调节性T细胞比例增高造成的免疫抑制可能在梅毒血清固定的发生中起着一定的作用。

沙眼衣原体感染可以引起多种疾病.而持续性和复发性是沙眼衣原体感染疾病的一个重要特点,了解导致其持续性的原因对于沙眼衣原体疾病的治疗,减少后遗症有重要作用.就近年来的一些相关研究,简要介绍了干扰素γ抑制被感染细胞凋亡,肿瘤坏死因子-α诱导T细胞凋亡,色氨酸合成酶的不同等几方面可能导致其持续性的原因.