Laboratory Investigation最新文献

Scoring Reliability of c-Met Immunohistochemical Assays in Lung Adenocarcinoma 肺腺癌c-Met免疫组化检测评分的可靠性。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-04-01 Epub Date: 2026-02-04 DOI: 10.1016/j.labinv.2026.106078

Yang Zhou , Junyi Pang , Yumeng Cai , Longyun Chen , Yinbo Xiao , Yining Zhen , Xianlong Chen , Zhiyong Liang , Xiaohua Shi

c-Met overexpression may identify patients who benefit from MET inhibitor therapy. However, concordance among c-Met immunohistochemistry (IHC) assays and scoring system reliability remain unclear. We retrospectively analyzed 150 lung adenocarcinoma specimens (from December 2018 to March 2023) using 5 c-Met IHC assays (SP44_R, SP44_Z, D1C2, LBP4-C-MET, and 811B7F4), scored by H-score, clinical score, 2-tier H-score (H-score ≥150 as positive) and 2-tier clinical score (clinical score 2+/3+ as positive). Next-generation sequencing for MET alterations was performed on 107 samples. H-scores of the 5 assays showed excellent interassay reliability (intraclass correlation coefficient = 0.953). Compared with SP44_R, which is the most frequently used antibody in the clinical trial, SP44_Z had the highest correlation with it (ρ = 0.851), followed by D1C2 (ρ = 0.818), LBP4-C-MET (ρ = 0.786), and 811B7F4 (ρ = 0.737). Among the 5 assays, Kendall

W

for the clinical score was 0.697; Fleiss κ for the 2-tier H-score was 0.51 and for the 2-tier clinical score was 0.494. In comparison with the clinical score 3+ of SP44_R, SP44_Z and 811B7F4 showed moderate reliability (κ = 0.483), LBP4-C-MET (κ = 0.459), and D1C2 (κ = 0.234). Nearly perfect agreement existed between the 2-tier H-score and clinical score (κ range: 0.944-0.986). In contrast, correlation analysis between H-scores across all assays and RNA expression levels revealed a weak association (ρ = 0.159-0.349). Five c-Met IHC assays demonstrated moderate-to-strong concordance in detecting c-Met overexpression. SP44_R, SP44_Z, LBP4-C-MET, and 811B7F4 performed reliably, although D1C2 was less consistent for clinical score 3+. A clinical score of 2+/3+ or an H-score of ≥150 is associated with high diagnostic consistency, supporting multiple validated IHC assays for c-Met evaluation in lung adenocarcinoma.

c-Met过表达可以识别从MET抑制剂治疗中获益的患者。然而，c-Met免疫组化（IHC）检测与评分系统可靠性之间的一致性尚不清楚。我们回顾性分析了150例肺腺癌标本（2018年12月至2023年3月），采用5种c-Met IHC检测（SP44R、SP44Z、D1C2、LBP4-C-MET和811B7F4），按H评分、临床评分、2级分类（H评分≥150或临床评分2+/3+为阳性）进行评分。对107个样本进行MET改变的下一代测序。5种检测方法的H评分具有良好的检测间信度（类内相关系数=0.953）。与临床试验中最常用的抗体SP44R相比，SP44Z与其相关性最高（ρ=0.851），其次为D1C2 （ρ=0.818）、LBP4-C-MET （ρ=0.786）、811B7F4 （ρ=0.737）。5项试验中，临床评分的Kendall’s W为0.697；两级H评分的Fleiss' κ为0.51，两级临床评分一致性为0.494。与SP44R临床评分3+相比，SP44Z和811B7F4具有中等信度（κ=0.483）， LBP4-C-MET (κ=0.459), D1C2 （κ=0.234）。两层H评分与临床评分几乎完全吻合（κ范围：0.944 ~ 0.986）。相比之下，所有检测的H评分与RNA表达水平之间的相关性分析显示弱相关性（ρ=0.159-0.349）。五种c-Met免疫组化检测在检测c-Met过表达方面表现出中等到强的一致性。SP44R、SP44Z、LBP4-C-MET和811B7F4表现可靠，但D1C2在临床评分3+时一致性较差。临床评分2+/3+或H评分≥150与高诊断一致性相关，支持多次验证的免疫组化检测用于肺腺癌c-Met评估。

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引用次数: 0

General Pathologists Achieve Near-Specialist Diagnostic Performance Using Deep Learning–Based Virtual Staining for Donor Kidney Assessment: A Retrospective-Prospective Diagnostic Concordance Study 普通病理学家使用基于深度学习的虚拟染色来评估供肾：一项回顾性-前瞻性诊断一致性研究。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-04-01 Epub Date: 2026-02-02 DOI: 10.1016/j.labinv.2026.106077

Jin-Peng Cen , Sheng-Dong Ge , Yang-Shu Zhou , Yu-Zhu Li , Ze-Feng Guo , Rong Peng , Yong-Guang Liu , Song Zhou , Shuo-Yu Xu , Shan-Chao Zhao , Ding Liu

Conventional rapid evaluation of donor kidney quality primarily relies on hematoxylin and eosin (H&E) staining, which inadequately visualizes collagen fibers, compromising the assessment of interstitial fibrosis and posing challenges for general pathologists (GPs) lacking specialized renal training. This study investigated whether artificial intelligence–based virtual staining could enhance donor kidney evaluation, particularly for interstitial fibrosis and chronic pathologies. Using 187 paired whole-slide images of H&E and Masson’s trichrome–stained sections, we developed and validated a CycleGAN-based virtual staining model that transforms H&E images into virtual Masson’s trichrome (VMT) representations. A renal pathologist (RP) and 2 GPs (GP.1 and GP.2) evaluated interstitial fibrosis and chronic changes using the Remuzzi scoring system, comparing results with and without virtual staining. Prospective validation was performed on 46 frozen sections. Results demonstrated that VMT effectively visualized interstitial collagen fibers, enabling more reliable fibrosis classification. Diagnostic accuracy significantly improved with virtual staining versus H&E alone (weighted kappa: GPs improved from 0.39-0.51 to 0.78-0.82; RP from 0.61 to 0.86), and interobserver agreement markedly increased (GPs: 55%-64% to 84%-88%; RP: 71%-90%). Notably, VMT bridged the expertise gap, allowing GPs to achieve near-specialist performance in fibrosis assessment (weighted kappa: GP.1 from 0.48 to 0.82; GP.2 from 0.41 to 0.84). Prospective validation confirmed these advantages, showing superior accuracy when combining VMT with H&E versus H&E alone (weighted kappa: GP.1 from 0.27 to 0.65; GP.2 from 0.34 to 0.68). The technology also enhanced glomerulosclerosis detection (GPs’ kappa = 0.82-0.87) and transplant decision-making accuracy (kappa = 0.84-0.85), elevating GPs to specialist-level competence. In conclusion, deep learning–based virtual staining significantly improves the precision of donor kidney evaluations by GPs, approaching specialist-level performance. This technology offers an efficient, cost-effective solution for assessing fibrosis and chronic pathologies, potentially eliminating diagnostic disparities between GPs and specialist pathologists in transplantation medicine.

传统的快速评估供肾质量主要依赖苏木精和伊红（H&E）染色，不能充分显示胶原纤维，影响间质纤维化的评估，并给缺乏专业肾脏培训的普通病理学家带来挑战。这项研究调查了基于人工智能的虚拟染色是否可以增强供体肾脏的评估，特别是对间质纤维化和慢性病理的评估。使用187对H&E和Masson三色染色切片的整片图像，我们开发并验证了基于cyclegan的虚拟染色模型，该模型将H&E图像转换为虚拟Masson三色（VMT）表示。一名肾脏病理学家（RP）和两名普通病理学家（GP.1和GP.2）使用Remuzzi评分系统评估间质纤维化和慢性变化，比较有无虚拟染色的结果。对46个冷冻切片进行前瞻性验证。结果表明，VMT能有效地显示间质胶原纤维，从而实现更可靠的纤维化分类。与单独使用H&E相比，虚拟染色显著提高了诊断准确性（加权kappa: gp从0.39-0.51提高到0.78-0.82；RP从0.61提高到0.86），观察者间一致性显著提高（gp从55-64%提高到84-88%；RP从71%提高到90%）。值得注意的是，VMT弥补了专业知识的差距，使全科医生在纤维化评估中达到接近专家的表现（加权κ: GP.1从0.48到0.82；GP.2从0.41到0.84）。前瞻性验证证实了这些优势，VMT联合H&E比单独H&E显示出更高的准确性（加权κ: GP.1从0.27到0.65；GP.2从0.34到0.68）。该技术还提高了肾小球硬化的检测（GPs′κ=0.82-0.87）和移植决策的准确性（κ=0.84-0.85），将普通病理学家提升到专家水平。总之，基于深度学习的虚拟染色显着提高了普通病理学家对供体肾脏评估的准确性，接近专家水平的表现。这项技术为评估纤维化和慢性病理提供了一种高效、经济的解决方案，有可能消除移植医学中普通病理学家和专科病理学家之间的诊断差异。

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引用次数: 0

Atypical Ductular Reactions Are a Distinct Regenerative Phenomenon in Hepatocellular Carcinoma (HCC) Patients after Transarterial Chemoembolization. 非典型导管反应是肝细胞癌（HCC）患者经动脉化疗栓塞后的一种独特的再生现象。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-19 DOI: 10.1016/j.labinv.2026.106122

Ornella Wolff, Adrian Kobe, Laure-Alix Clerbaux, Christoph Bütikofer, Magdalene Adamczyk, Markus Rechsteiner, Achim Weber, Juliane Friemel

Transarterial chemoembolization (TACE) is increasingly used in patients with hepatocellular carcinoma (HCC), the most frequent primary liver tumor. Atypical ductular reactions challenge the histological diagnosis in specimens of patients after embolization treatment, mimicking intrahepatic cholangiocarcinoma (CCA) or combined hepatocellular cholangiocarcinoma (HCC-CCA). We characterized atypical ductular reactions in HCC patients after embolization treatment in association with therapy mode, response and survival using histomorphology and spatial microdissection. This retrospective, single-center, longitudinal cohort study included 188 HCC patients. We compared a study group (n=105 patients with total 206 tumor nodules) vs. a treatment-naïve control group (n=83/112 nodules). Presence of embolization compounds, type of embolization treatment, tumor necrosis, morphology and the expression of biliary markers (CK7/19, SOX9) were assessed on whole slides and a tissue microarray (TMA) using machine learning assisted marker profiling. Spatial microdissection and sequencing with the Oncomine Comprehensive Assay (OCAv3) tested genetic landscapes. The relative risk for atypical ductular reactions was increased in HCC patients following embolization therapy compared to the control group (RR 3.6; p<0.05). Morphology appears as ribbon-like with atypical, confluent ducts usually found on the interface of necrosis and vital tumor nodules. Strong intensity of CK7/19 and SOX9 was observed throughout, compared to a heterogenous expression pattern in combined HCC-CCA. Spatial microdissection of atypical ductular reactions showed variants of unknown significance in 1/13 cases (NTRK3), but no CCA-typical mutations. In contrast, combined HCC-CCA (n=4) displayed oncogene alterations in known targets (TP53, BAP1 and TERT). Increased atypical ductular reactions in specimens of HCC-patients after transarterial embolization treatment can be identified as regenerative phenomenon with distinct morphology and immunophenotype.

经动脉化疗栓塞（TACE）越来越多地用于肝细胞癌（HCC）患者，HCC是最常见的原发性肝脏肿瘤。在栓塞治疗后的患者标本中，模拟肝内胆管癌（CCA）或肝细胞型胆管癌（HCC-CCA）的非典型导管反应挑战组织学诊断。我们利用组织形态学和空间显微解剖分析了HCC患者栓塞治疗后的非典型导管反应与治疗方式、反应和生存的关系。这项回顾性、单中心、纵向队列研究纳入188例HCC患者。我们比较了研究组（n=105例患者，共206个肿瘤结节）和treatment-naïve对照组（n=83/112个结节）。栓塞化合物的存在、栓塞治疗的类型、肿瘤坏死、形态学和胆道标志物（CK7/19、SOX9）的表达在整个载玻片和组织微阵列（TMA）上进行评估，使用机器学习辅助标志物分析。空间显微解剖和测序与Oncomine综合分析（OCAv3）测试遗传景观。与对照组相比，栓塞治疗后HCC患者发生非典型导管反应的相对风险增加(RR 3.6; p

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引用次数: 0

PD-L1 Testing in Triple-Negative Breast Cancer: National Practice and Interlaboratory Variation in the Netherlands. 三阴性乳腺癌的PD-L1检测：荷兰的国家实践和实验室间差异。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-13 DOI: 10.1016/j.labinv.2026.106121

Mirte Dekker, Maaike Anna Hempenius, Geertruida H de Bock, Michel van Kruchten, Marcel A T M van Vugt, Albert G Siebers, Bert van der Vegt

Purpose: This nationwide study aimed to evaluate Programmed Death-Ligand 1 (PD-L1) test characteristics and test results during the first two years after the introduction of PD-L1 testing in triple-negative breast cancer (TNBC), and to analyze interlaboratory variation in PD-L1 positivity rates, assessed by a combined positive score (CPS), using nationwide real-world data from the Netherlands.

Materials and methods: Pathology reports of TNBC patients mentioning PD-L1 were retrieved from the Dutch nationwide pathology database (Palga). PD-L1 test characteristics and results were evaluated per year between August 2022 and August 2024. To assess consistency between laboratories, interlaboratory variation in PD-L1 positivity rates (CPS≥10) was analyzed using funnel plots with 95% confidence intervals around the national mean. Laboratory specific PD-L1 positivity rates were case-mix adjusted, using multivariate logistic regression.

Results: During the study period, 926 PD-L1 tests were performed (423 in year 1 and 503 in year 2). CPS was applied in 94.5% of cases. The national mean PD-L1 positivity rate was 46.7%. After adjustment for case-mix variables, laboratory-specific positivity rates ranged from 21.3% to 70.7%, with three of the 19 laboratories showing significant deviations from the national mean.

Conclusions: PD-L1 testing for TNBC is generally well conducted according to the national guidelines in the Netherlands, with consistent use of CPS. However, the observed interlaboratory variation in positivity rates highlights the need for further standardization of testing methods to ensure accuracy of patient selection for immunotherapy.

目的：这项全国性的研究旨在评估三阴性乳腺癌（TNBC）引入PD-L1检测后头两年的程序性死亡配体1 （PD-L1）检测特征和检测结果，并分析PD-L1阳性率的实验室间差异，通过联合阳性评分（CPS）评估，使用来自荷兰的全国真实数据。材料和方法：从荷兰全国病理数据库（Palga）中检索涉及PD-L1的TNBC患者的病理报告。在2022年8月至2024年8月期间，每年评估PD-L1测试特征和结果。为了评估实验室之间的一致性，使用漏斗图分析PD-L1阳性率（CPS≥10）的实验室间差异，其95%置信区间在全国平均值附近。实验室特异性PD-L1阳性率采用多因素logistic回归进行病例混合校正。结果：在研究期间，进行了926次PD-L1测试（第1年423次，第2年503次）。94.5%的病例采用CPS。全国PD-L1平均阳性率为46.7%。在对病例组合变量进行调整后，实验室特异性阳性率从21.3%到70.7%不等，19个实验室中有3个与全国平均水平存在显著偏差。结论：根据荷兰的国家指南，在一贯使用CPS的情况下，TNBC的PD-L1检测通常进行得很好。然而，观察到的实验室间阳性率差异强调了进一步标准化检测方法的必要性，以确保患者选择免疫治疗的准确性。

{"title":"PD-L1 Testing in Triple-Negative Breast Cancer: National Practice and Interlaboratory Variation in the Netherlands.","authors":"Mirte Dekker, Maaike Anna Hempenius, Geertruida H de Bock, Michel van Kruchten, Marcel A T M van Vugt, Albert G Siebers, Bert van der Vegt","doi":"10.1016/j.labinv.2026.106121","DOIUrl":"https://doi.org/10.1016/j.labinv.2026.106121","url":null,"abstract":"<p><strong>Purpose: </strong>This nationwide study aimed to evaluate Programmed Death-Ligand 1 (PD-L1) test characteristics and test results during the first two years after the introduction of PD-L1 testing in triple-negative breast cancer (TNBC), and to analyze interlaboratory variation in PD-L1 positivity rates, assessed by a combined positive score (CPS), using nationwide real-world data from the Netherlands.</p><p><strong>Materials and methods: </strong>Pathology reports of TNBC patients mentioning PD-L1 were retrieved from the Dutch nationwide pathology database (Palga). PD-L1 test characteristics and results were evaluated per year between August 2022 and August 2024. To assess consistency between laboratories, interlaboratory variation in PD-L1 positivity rates (CPS≥10) was analyzed using funnel plots with 95% confidence intervals around the national mean. Laboratory specific PD-L1 positivity rates were case-mix adjusted, using multivariate logistic regression.</p><p><strong>Results: </strong>During the study period, 926 PD-L1 tests were performed (423 in year 1 and 503 in year 2). CPS was applied in 94.5% of cases. The national mean PD-L1 positivity rate was 46.7%. After adjustment for case-mix variables, laboratory-specific positivity rates ranged from 21.3% to 70.7%, with three of the 19 laboratories showing significant deviations from the national mean.</p><p><strong>Conclusions: </strong>PD-L1 testing for TNBC is generally well conducted according to the national guidelines in the Netherlands, with consistent use of CPS. However, the observed interlaboratory variation in positivity rates highlights the need for further standardization of testing methods to ensure accuracy of patient selection for immunotherapy.</p>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":" ","pages":"106121"},"PeriodicalIF":4.2,"publicationDate":"2026-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147463687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Diagnostic Potential of Tn-MUC1 in Breast Cancer: A Novel Immunohistochemical Marker Reflecting Tumor Progression n- muc1在乳腺癌中的诊断潜力：一种反映肿瘤进展的新型免疫组织化学标志物。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-01-06 DOI: 10.1016/j.labinv.2025.106070

Ai Shimizu , Kanako C. Hatanaka , Ayae Nange , Asami Okumura , Masanori Takehashi , Yoshiki Shinomiya , Kentaro Naruchi , Masaharu Sato , Hiroshi Kase , Tomoko Mitsuhashi , Hiroko Yamashita , Yutaka Hatanaka , Yoshihiro Matsuno

Mucin 1 (MUC1) is a highly O-glycosylated transmembrane glycoprotein. Tumor-associated MUC1, characterized by aberrant O-linked glycans, is overexpressed in cancer cells; however, conventional MUC1 antibodies show limited specificity for tumor-associated glycan structures. Recently, a novel epitope-defined antibody (MUC1-Tn antigen epitope-defined antibody [MUC1-Tn ED Ab]) that specifically recognizes the Tn-MUC1 antigen was developed. In this study, we evaluated the potential of MUC1-Tn ED Abs as diagnostic markers of breast cancer. Tissue microarray sections from 124 patients with invasive breast carcinoma (IBC) and 26 whole tissue sections, including multiple neoplastic lesions—flat epithelial atypia (n = 24), ductal carcinoma in situ (n = 26), and IBC (n = 16)—were analyzed. Immunohistochemical distributions of Tn-MUC1 and MUC1 were assessed using the MUC1-Tn ED Ab (clone SN102) and a conventional antibody (clone Ma552), respectively. In tissue microarray analysis, Tn-MUC1 exhibited minimal immunoreactivity in nonneoplastic areas and high specificity for IBC. In IBC tissues, immunoreactivity with Tn-MUC1 was predominantly cytoplasmic, unlike conventional MUC1 staining observed in both the cytoplasm and membrane. In multilesion analysis, cytoplasmic Tn-MUC1 expression was rarely detected in nonneoplastic areas but progressively increased across flat epithelial atypia, ductal carcinoma in situ, and IBC. Knockdown assays in breast cancer cell lines demonstrated that core 1 β1,3-galactosyltransferase 1 (C1GALT1), an enzyme involved in galactosylation of the Tn antigen, significantly influenced the cellular localization of Tn-MUC1. This study demonstrates that Tn-MUC1, as detected by the MUC1-Tn ED Ab, has high specificity for breast cancer and may act as a novel immunohistochemical marker reflecting tumor progression.

Mucin 1 （MUC1）是一种高度o糖基化的跨膜糖蛋白。肿瘤相关（TA） MUC1 (TA-MUC1)，以异常的o链聚糖为特征，在癌细胞中过度表达；然而，传统的MUC1抗体对TA聚糖结构的特异性有限。最近，一种特异性识别n- muc1抗原的新型表位定义抗体（MUC1-Tn ED Ab）被开发出来。在这项研究中，我们评估了MUC1-Tn ED抗体作为乳腺癌诊断标志物的潜力。我们分析了124例浸润性乳腺癌（IBC）患者的组织微阵列（TMA）切片和26例全组织切片，包括多发性肿瘤病变-扁平上皮异型性（FEA, n = 24）、导管原位癌（DCIS, n = 26）和IBC （n = 16）。使用MUC1- tn ED Ab（克隆SN102）和常规抗体（克隆Ma552）分别评估Tn-MUC1和MUC1的免疫组化分布。在TMA分析中，n- muc1在非肿瘤区域表现出最小的免疫反应性，对IBC具有高特异性。在IBC组织中，与n-MUC1的免疫反应性主要发生在细胞质中，这与在细胞质和细胞膜中观察到的常规MUC1染色不同。在多病变分析中，细胞质n- muc1表达在非肿瘤区域很少检测到，但在FEA， DCIS和IBC中逐渐增加。乳腺癌细胞系的敲低实验表明，参与Tn抗原半乳糖基化的酶core 1 β1,3-半乳糖基转移酶1 （C1GALT1）显著影响Tn- muc1的细胞定位。本研究表明，通过MUC1-Tn ED Ab检测到的n- muc1对乳腺癌具有高特异性，可能作为一种反映肿瘤进展的新型免疫组织化学标志物。

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引用次数: 0

Differential Expression of P-Element–Induced Wimpy Testis (PIWI)–Interacting RNAs in Glioblastoma Stem Cells Affects Their Biological Features: Implications for Tumor Progression and Patient Survival 胶质母细胞瘤干细胞中piwi相互作用rna的差异表达影响其生物学特性：对肿瘤进展和患者生存的影响

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-01-13 DOI: 10.1016/j.labinv.2026.106072

Frantisek Siegl , Elena Garcia-Borja , Rosana Mateu , Karolina Trachtova , Matej Jasik , Martin Kopecky , Marek Vecera , Lenka Radova , Michal Hendrych , Tomas Kazda , Pavel Fadrus , Aleksi Sedo , Ondrej Slaby , Petr Busek , Jiri Sana

Glioblastoma isocitrate dehydrogenase–wild-type (GBM) is the most lethal and, concurrently, the most common malignant primary brain tumor. Its aggressive behavior is associated with the presence of a subpopulation of glioblastoma stem cells (GSCs) that drive resistance to therapy and early relapse. P-element–induced wimpy testis–interacting RNAs (piRNAs), which are characteristically expressed in undifferentiated cells, are thought to play an important role in GSC biology. To identify piRNAs associated with GSCs, we selected 24 paired GSC and non-GSC cultures from patients with GBM based on the expression of stem cell markers CD133 and Sox2, ability of GSCs to form spheres in culture, and to differentiate and replicate tumors in immunodeficient mice. Global piRNA profiling using next-generation sequencing revealed 98 significantly differentially expressed piRNAs in GSCs compared with non-GSCs, including piR-9491, whose dysregulation in GBM has been previously described. Downregulation of piR-9491 in GSCs led to decreased viability, growth, invasion, and increased apoptosis. Significance of piRNA pathway for GBM pathology was further highlighted by identification of 34 piRNAs connected to patients' survival. These molecules were further used for the establishment of a piRNA signature predicting survival of patients with GBM. Our results not only confirm the important role of piR-9491 in GSCs but also indicate a potential significant role of piRNAs in the biology of GBM.

野生型胶质母细胞瘤（GBM）是最致命的，同时也是最常见的恶性原发性脑肿瘤。其侵袭性行为与胶质母细胞瘤干细胞（GSC）亚群的存在有关，该亚群驱动对治疗的抵抗和早期复发。piwi相互作用rna （piRNAs）在未分化细胞中特征性表达，被认为在GSC生物学中起重要作用。为了鉴定与GSC相关的pirna，我们基于干细胞标记物CD133和Sox2的表达、GSC在培养物中形成球的能力、免疫缺陷小鼠中分化和复制肿瘤的能力，从GBM患者中选择了24对GSC和非GSC培养物。使用下一代测序的全球piRNA分析显示，与非GSCs相比，GSCs中有98种piRNA表达显著差异，包括piR-9491，其在GBM中的失调先前已被描述过。piR-9491在GSCs中的下调导致细胞活力、生长、侵袭降低和细胞凋亡增加。通过鉴定34个与患者生存相关的piRNA，进一步强调了piRNA通路在GBM病理中的意义。这些分子进一步用于建立预测GBM患者生存的piRNA标记。我们的研究结果不仅证实了piR-9491在GSCs中的重要作用，而且还表明了pirna在GBM生物学中的潜在重要作用。

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引用次数: 0

Clonal Progression of a DNA Polymerase Epsilon–Mutant Endometrial Cancer With Immune Evasion Characteristics and Metastasis 具有免疫逃避特征和转移的DNA聚合酶epsilon突变子宫内膜癌的克隆进展

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-01-20 DOI: 10.1016/j.labinv.2026.106074

Ruirui Fan , Jie Gao

Renal transplant recipients are at significantly higher risk of developing malignancies because of chronic immunosuppression. Although DNA polymerase epsilon (POLE)–mutated endometrioid carcinoma (EC) typically presents as early-stage disease with an excellent prognosis and rare metastasis, we conducted a study on POLE-mutated EC in a transplant recipient that rapidly progressed and metastasized. We aimed to first elucidate the molecular and immunological mechanisms of rapid and malignant progression in the tumor. To understand the mechanisms underlying this atypical clinical course, we performed histological and immunohistochemical comparisons of the primary endometrial tumor and metastatic peritoneal lesions, alongside next-generation sequencing and RNA sequencing analyses. The primary and metastatic lesions shared 17 somatic mutations, indicating a common origin but demonstrated divergent evolution. Gene expression profiling revealed low CD8+ T-cell infiltration and weak interferon gamma signaling, suggesting an immunologically inactive or “immune desert” tumor microenvironment. Clonal evolution analysis showed that the POLE p.S297F mutation initiated hypermutation and clonal diversification in the primary tumor. In contrast, metastatic lesions exhibited POLE loss and activation of the BRAF V600E::PIK3CD pathway, leading to progress and immune evasion. These findings highlight a distinct evolutionary trajectory of POLE-mutated EC under immunosuppressive conditions in this case and underscore the need for tailored oncological surveillance and treatment strategies in transplant recipients.

由于慢性免疫抑制，肾移植受者发生恶性肿瘤的风险明显更高。虽然DNA聚合酶（POLE）突变的子宫内膜样癌（EC）通常表现为早期疾病，预后良好，很少转移，但我们在移植受体中进行了一项pol突变的EC的研究，该EC迅速进展和转移。我们的目的是首先阐明肿瘤快速和恶性进展的分子和免疫学机制。为了了解这种非典型临床病程的机制，我们对原发性子宫内膜肿瘤和转移性腹膜病变进行了组织学和免疫组织化学比较，并进行了下一代测序和rna测序分析。原发性和转移性病变共有17个体细胞突变，表明有共同的起源，但表现出不同的进化。基因表达谱显示低CD8+ T细胞浸润和弱干扰素γ信号，提示免疫无活性或“免疫荒漠”肿瘤微环境。克隆进化分析表明，突变POLE p.S297F在原发肿瘤中引发了高突变和克隆多样化。相反，转移性病变表现为POLE丢失和BRAF V600E/PIK3CD通路的激活，导致进展和免疫逃避。这些发现强调了在这种情况下免疫抑制条件下pole突变EC的独特进化轨迹，并强调了在移植受者中定制肿瘤监测和治疗策略的必要性。

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引用次数: 0

Retraction notice to "Extracellular cold-inducible RNA-binding protein regulates neutrophil extracellular trap formation and tissue damage in acute pancreatitis" Lab Invest 2020 Dec;100(12):1618-1630 “细胞外冷诱导rna结合蛋白调节急性胰腺炎中性粒细胞胞外陷阱形成和组织损伤”的撤回通知，中国医学杂志2020年12月；100(12):1618-1630。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-02-17 DOI: 10.1016/j.labinv.2026.106076

Johan Linders , Raed Madhi , Milladur Rahman , Matthias Mörgelin , Sara Regner , Max Brenner , Ping Wang , Henrik Thorlacius

引用次数: 0

The Value of Internal Positive Controls and Synthetic Calibrators for Performance Assessment of Breast Carcinoma Estrogen Receptor Immunohistochemistry Assay 内阳性对照和合成校准器对乳腺癌雌激素受体免疫组化检测性能评价的价值。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-01-06 DOI: 10.1016/j.labinv.2025.106065

Emina E. Torlakovic , Seshi R. Sompuram , Søren Nielsen , Steven A. Bogen

In 2010, the American Society of Clinical Oncology and the College of American Pathologists jointly issued guidelines for estrogen receptor (ER) and progesterone receptor testing and an update in 2020. Both guidelines recommended that ER and progesterone receptor assays be interpreted in the context of internal and external controls. Despite the considerable importance of internal controls for ER testing by immunohistochemistry (IHC), there are no studies that assess their performance and effectiveness. The Canadian Immunohistochemistry Quality Control performed an educational proficiency testing run for 70 laboratories using a tissue microarray with 80 samples of breast carcinoma enriched for tumors weakly positive for ER. The tissue microarray contained several internal positive controls and was run together with ER calibrators from Boston Cell Standards. The internal positive controls were scored as negative in 0/70 (0%), weak in 12/70 (17%), moderate in 29/70 (41%), and strong in 29/70 (41%) laboratories secondary to differences in analytical sensitivity of IHC protocols. For calibrators, only SP1 clone results (52 participants) are reported because of the small number of laboratories using other primary antibody clones. Thirty-nine laboratories (75%) had IHC protocols with (limit of detection) LOD < 25,000. Both internal positive controls and Boston Cell Standard ER calibrators showed excellent correlation with overall H-score mean, percent 3+ positive cells, and percent negative cells (P < .0001). The difference in diagnostic sensitivity was significant between weak and moderate and between weak and strong internal positive controls (t test, P = .015 and P = .009, respectively) and between IHC protocols with LOD < 25,000 and all others (t test, P = .001). The measured LOD had a significant association with diagnostic accuracy at 1% and 10% cutoffs, whereas the internal positive control score was significantly associated only with diagnostic accuracy at the 10% cutoff (1-way analysis of variance, P = .009). Negative results of the ER IHC assay should not be reported when internal positive controls are either negative or weak. Further studies with more advanced calibrators are needed in order to establish the recommended LOD range for optimal ER IHC assay performance with SP1 and other anti-ER primary antibody clones.

2010年，美国临床肿瘤学会（ASCO）和美国病理学家学会（CAP）联合发布了雌激素受体（ER）和孕激素受体（PgR）检测指南，并将于2020年更新。两份指南都建议在内部和外部控制的背景下解释ER和PgR测定。尽管通过免疫组织化学（IHC）进行内源性雌激素检测的内部控制相当重要，但没有研究评估其性能和有效性。加拿大免疫组织化学质量控制中心（CIQC）使用组织微阵列（TMA）对70个实验室进行了教育水平测试，其中80个乳腺癌样本富含ER弱阳性肿瘤。TMA包含几个内部阳性对照，并与来自波士顿细胞标准公司（BCS）的ER校准器一起运行。由于免疫组化方案分析敏感性的差异，内部阳性对照在0/70（0%）中评分为阴性，在12/70（17%）中评分为弱，在29/70（41%）中评分为中等，在29/70（41%）中评分为强。由于使用其他一抗克隆的实验室数量较少，校准者只报告了SP1克隆结果（52名参与者）。39个实验室（75%）的IHC方案LOD < 25,000。内部阳性对照和BCS ER校准器与总体h评分平均值、3+阳性细胞百分比和阴性细胞百分比均表现出极好的相关性（p < 0.0001）。诊断敏感性在弱和中度、弱和强内部阳性对照之间存在显著差异（t检验，p分别= 0.015和p = 0.009），在LOD < 25,000的IHC方案和所有其他方案之间存在显著差异（t检验，p = 0.001）。测量的LOD与1%和10%临界值的诊断准确性显著相关，而内部阳性控制评分仅与10%临界值的诊断准确性显著相关（单向方差分析，p = 0.009）。当内部阳性对照阴性或弱时，不应报告ER免疫组化试验的阴性结果。需要使用更先进的校准器进行进一步的研究，以确定SP1和其他抗ER一抗克隆的最佳ER IHC检测性能的推荐LOD范围。

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引用次数: 0

Optimal Formalin-Fixed Paraffin-Embedded Sample Preservation for Efficient Staining in Multiplex Imaging 福尔马林固定石蜡包埋样品在多重成像中高效染色的最佳保存方法。

IF 4.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2026-03-01 Epub Date: 2026-01-22 DOI: 10.1016/j.labinv.2026.106075

Yuna Delarue , Nadège Marec , Marion Le Rochais , Christelle Le Dantec , Soizic Garaud , Arnaud Uguen , Divi Cornec , Christophe Jamin , Pierre Pochard , Patrice Hemon

The accuracy of immunohistochemical techniques depends critically on the preservation of antigenic integrity in tissue samples. Although formalin-fixed paraffin-embedded tissues are commonly used for long-term storage, several studies reported signal degradation over time. However, the extent of this degradation and the influence of storage conditions on a wide range of markers remain poorly characterized. In this study, tissue blocks collected over an 8-year period and tissue sections stored under different conditions (room temperature [RT], 4 °C, −20 °C, and −80 °C) were analyzed. Twenty-five antibodies targeting immune, stromal, and structural markers were used to stain 4 different tissue sections. Samples were acquired using imaging mass cytometry. Signal intensities were first quantified, then the data were analyzed, before and after normalization, using the imcRtools pipeline. A gradual loss of signal intensity was observed in blocks stored at RT for more than 6 years, affecting the detection of certain markers regardless of tissue type. For some sections, significant signal degradation was observed after only 1 week at RT and for others after 1 month of storage. Markers such as CD20, CD45, and CD45RA proved to be particularly sensitive. Storage at −20 °C or −80 °C preserved the quality of the staining, whereas storage at 4 °C only allowed partial preservation. Normalization corrected some variations but did not enable the recovery of signals that were severely altered or absent. Commonly used RT storage causes a gradual reduction in imaging mass cytometry--detected immunoreactivity, which may affect scientific interpretation and diagnosis. Storing sections at −20 °C is, therefore, an effective and accessible solution. For retrospective studies, we recommend using blocks that are less than 6 years old. Overall, these results highlight the importance of preestablished standardized preservation protocols to ensure the reproducibility of analyses.

免疫组织化学技术的准确性主要依赖于组织样品中抗原完整性的保存。虽然福尔马林固定石蜡包埋组织通常用于长期储存，但一些研究报告信号会随着时间的推移而退化。然而，这种降解的程度和储存条件对各种标记物的影响仍然不清楚。在本研究中，我们分析了在8年期间收集的组织块和在不同条件下（室温，4°C, -20°C, -80°C）保存的组织切片。使用25种针对免疫、基质和结构标记物的抗体对四个不同的组织切片进行染色。采用成像细胞术（IMC）采集样本。首先对信号强度进行量化，然后使用imcRtools流水线对归一化前后的数据进行分析。在室温下储存6年以上，观察到信号强度逐渐丧失，影响了某些标记物的检测，无论组织类型如何。对于一些切片，在室温下仅一周就观察到明显的信号退化，而另一些则在一个月后储存。CD20、CD45和CD45RA等标记物被证明是特别敏感的。储存在-20°C或-80°C保存染色的质量，而储存在4°C只能部分保存。归一化纠正了一些变化，但不能恢复严重改变或缺失的信号。常用的室温保存方法会导致imc检测到的免疫反应性逐渐降低，这可能会影响科学解释和诊断。因此，将切片保存在-20°C是一种有效且易于使用的解决方案。对于回顾性研究，我们建议使用年龄小于6岁的积木。总的来说，这些结果强调了预先建立标准化保存协议以确保分析可重复性的重要性。

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引用次数: 0