目的 探讨甲状腺乳头状癌术前细针穿刺细胞学(fine-needle aspiration cytology,FNAC)和术后石蜡组织BRAF V600E突变检测的差异及应用价值.方法 应用免疫组化和qRT-PCR检测143例甲状腺乳头状癌石蜡标本BRAF V600E突变,以测序为金标准,比较两种技术检测BRAF V600E的差异.回顾分析265例甲状腺乳头状癌术前FNAC和术后石蜡组织采用qRT-PCR检测BRAF V600E,比较同一组织不同获取方式对BRAF V600E突变检出率的影响.结果 143例甲状腺乳头状癌石蜡标本检测BRAF V600E,免疫组化特异度为85.45％(47/55)、阳性预测值为91.67％(88/96)、准确率为94.41％(135/143)、假阳性率为14.55％(8/55).qRT-PCR特异度为98.19％(54/55)、阳性预测值为98.88％(88/89)、准确率为99.30％(142/143)、假阳性率为1.8％(1/55).经配对χ2检验,比较两种方法检测的差异,其特异度、阳性预测值、准确率、假阳性率差异有显著性(P<0.00001).另外,265例甲状腺乳头状癌患者术前FNAC BRAF V600E突变检出率为69.81％(185/265),术后石蜡组织BRAF V600E突变检出率为76.98％(204/265).经配对χ2检验,术前FNAC和术后石蜡组织BRAF V600E突变检出率差异有统计学意义(P<0.00001).结论 甲状腺乳头状癌采用qRT-PCR检测BRAF V600E突变优于免疫组化;术后石蜡组织BRAF V600E突变检出率明显高于术前FNAC标本,甲状腺乳头状癌术前FNAC检测BRAF V600E阴性的病例,建议术后石蜡组织复检BRAF V600E突变状态.

恶性间皮瘤(malignent mesothelioma,MM)是一种显示向间皮分化的恶性肿瘤[1],好发于胸膜和腹膜,少数病例可发生于心包膜和睾丸鞘膜等部位.呈弥漫性生长的称为弥漫性恶性间皮瘤(DMM),少数肿瘤(＜1％)可呈局限性结节性生长,称为局限性恶性间皮瘤(LMM).

Objective: The purpose of this study was to investigate the relationships between TP53 Pro72Arg (rs1042522) polymorphism and susceptibility to type 2 diabetes (T2DM) and its related complications. Methods: The TP53 Pro72Arg polymorphism was genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method in 206 T2DM patients and 446 healthy controls. Mitochondrial DNA (mtDNA) content, mtDNA transcriptional level and large-scale mtDNA deletion were evaluated in leukocytes of T2DM patients using fluorescence-based quantitative PCR (FQ-PCR), reverse transcriptase-quantitative PCR (RTqPCR) and long-range PCR approaches, respectively. The data of our study were processed by GraphPad Prism software (version 7.00). Results: The distribution of TP53 Pro72Arg differed in T2DM patients from the controls, with a moderately increased proportion of TP53 Arg72 variant carriers (Pro/Arg and Arg/Arg genotypes) (88.3% vs 81.2%, p=0.022; OR=1.089, 95% CI=1.018-1.164). T2DM patients with Arg/ Arg genotype had significantly decreased prevalences of diabetic neuropathy and retinopathy compared to those without (6.5% vs 19.4%, p=0.018 and 14.8% vs 30.7%, p=0.018, respectively). T2DM patients with Arg/Arg genotype had higher mtDNA content and mtRNA expression level than those who were not Arg/Arg genotype (p<0.05 for all), and we did not observe mtDNA 4977-base pair (bp) deletion mutations in the leukocytes of T2DM patients. Conclusion: There was a significant association of the TP53 Pro72Arg polymorphism with susceptibility to T2DM, and the homozygous Arg/Arg genotype of this gene locus might be a protective factor for diabetic complications. Those results suggested that the TP53 Arg72 variant had a different association with type 2 diabetes and its complications, and it might be related to mtDNA maintenance of the TP53 Arg72 variant under hyperglycemia-induced stress.

This study aims to identify differentially expressed proteins related with platinum sensitivity and to find biomarkers for predicting platinum response and survival outcomes in patients with high-grade serous ovarian cancer (HGSOC). Eligible HGSOC patients were divided into platinum-sensitive and platinum-resistant groups according to platinum-free interval (PFI). Tissue protein lysates from tumor tissues were subjected to an in-solution tryptic digest followed by tandem mass tag (TMT) labeling of the resulting peptides and mass spectrometric analysis. Candidate proteins were identified using differentially expressed protein and gene set enrichment analysis (GSEA) and confirmed by immunohistochemistry (IHC), and their survival relevance was evaluated in The Cancer Genome Atlas (TCGA) ovarian cancer cohort. The results showed that there was a significant difference in the protein expression profiling between the two patient groups. In the GSEA model, a gene set of 239 extracellular matrix (ECM)-related proteins was significantly enriched in the platinum-sensitive group [normalized enrichment score (NES) = 3.82, q < 10 −5 ], and this finding was confirmed in TCGA ovarian cancer cohort. Interestingly, an ECM-related gene expression, serpin family A member 10 ( SERPINA10 ), was identified to be significantly positively correlated with overall survival (OS) and progression-free survival (PFS) in TCGA ovarian cancer cohort (all p < 0.05). IHC results demonstrated that HGSOC patients with high SERPINA10 expression had longer PFI than the patients with low SERPINA10 expression (9 vs. 5 months, p = 0.038), and the SERPINA10 expression had an area under the receiver operating characteristic curve (AUC) value of 0.758 (95% CI = 0.612–0.905; p = 0.005) to discriminate the platinum-sensitive group from the platinum-resistant group. In conclusion, the results suggested that SERPINA10 could be a promising biomarker for predicting the response and survival in platinum-based chemotherapy of HGSOC.

目的 应用免疫组化和实时荧光定量PCR(real-time fluorescence quantitative PCR,qRT-PCR)技术检测结直肠癌中BRAF V600E突变,比较这两种技术在结直肠癌中的应用价值.方法 总结113例结直肠癌标本BRAF V600E免疫组化和qRT-PCR的结果 ,以测序为金标准,评价这两种技术在结直肠癌BRAF V600E突变检测中的应用价值.结果 113例结直肠癌标本中,免疫组化检测阳性19例,阴性82例,不确定诊断12例.其中19例阳性标本经测序确认BRAF V600E突变6例,未突变13例;12例不确定诊断病例经测序确认1例突变,11例未突变.免疫组化阳性检出率为16.81％,阴性检出率为72.57％,不确定诊断例数占检测例数的10.62％.113例结直肠癌标本同时行qRT-PCR检测,结果 突变7例,未突变106例,阳性检出率为6.19％,阴性检出率为93.8％,无不确定诊断,与测序结果 一致,符合率达100％.以测序为金标准,比较这两种技术在结直肠癌检测BRAF V600E中的应用价值,免疫组化和qRT-PCR技术的灵敏度、特异度、阳性预测值、阴性预测值、假阴性率、假阳性率、准确率分别为100％、86.32％、31.58％、100％、0、13.68％、87.13％和100％、100％、100％、100％、0、0、100％.经配对χ2检验这两种检测方法差异有显著性(P<0.00001).结论 在结直肠癌BRAF V600E检测中,qRT-PCR较免疫组化有更高的准确性.

Purpose Platinum resistance is a primary barrier to improving the survival rate of ovarian cancer. The relationship between mtDNA somatic mutations and response to platinum-based chemotherapy in ovarian cancer has not been well clarified. Patients and Methods Here, we employed the next-generation sequencing (NGS) platform to identify mtDNA mutations of the unrelated high-grade serous ovarian cancer (HGSOC) patients. Results We identified 569 germline variants and 28 mtDNA somatic mutations, and found the platinum-sensitive relapsed HGSOC patients had more synonymous mutations while the platinum-resistant relapsed HGSOC patients had more missense mutations in the mtDNA somatic mutations. Meanwhile, we found that the HGSOC patients who harbored heteroplasmic pathogenic mtDNA somatic mutations had significantly higher prevalence of both platinum-resistance and relapse than those without (80.0% versus 16.7%, p=0.035). Additionally, we observed that the tumor tissues had significantly higher lactate-to-pyruvate (L/P) ratio than the paired nontumor tissues (p<0.001), and L/P ratio of tumors with any heteroplasmic pathogenic mtDNA mutations was significantly higher than that of the tumors free of pathogenic mtDNA mutations (p=0.025). Conclusion Our findings indicate that these heteroplasmic pathogenic mtDNA somatic mutations may cause decreased respiratory chain activity and lead to the metabolism remodeling that seem to be beneficial for progression of both platinum-based chemotherapy resistance and relapse.

目的 探讨非典型官颈腺细胞(AGC)在宫颈细胞学中的应用情况.方法 2012年1月～2019年8月选取南京医科大学第二附属医院病理科经液基薄层细胞检测检查的AGC病例34例,与其组织学结果进行对照并分析.结果 宫颈腺上皮异型增生7例(20.59％),宫颈腺癌2例(5.88％),子宫内膜腺癌4例(11.76％),高度鳞状上皮内病变累及腺体9例(26.47％),低分化鳞状细胞癌2例(5.88％),宫颈管内膜慢性炎7例(20.59％),子宫内膜息肉2例(5.88％),宫颈腺上皮微腺型增生1例(2.94％).根据病变类型,分为腺上皮病变组(13例)和鳞状上皮病变组(11例),两组年龄构成比较,差异有统计学意义(P＜0.05).结论 AGC在官颈组织学对照中,若为良性或者癌前病变,多数为官颈管腺上皮异形增生以及较大比例的需要鉴别的高度鳞状上皮内病变累及腺体;若为恶性,则多数为宫颈腺癌或子宫内膜腺癌及需要鉴别的低分化鳞癌.此外,不同病变类型可能与年龄有关系,40岁以上是腺上皮病变的高发年龄段.

BACKGROUND:The pathogenesis of acquired melanocytic nevi (AMN) is still unclear, and the origin of nevus cells has not been clarified.OBJECTIVE:To analyze the clinical features and pathological types of AMN and identify the possible origin of nevus cells.METHODS:A retrospective study of 2929 cases of AMN was conducted, and 96 specimens of intradermal and junctional nevi were selected. Immunohistochemical assays were performed to detect the expression of basement membrane component receptor DDR-1 and the molecular markers on epidermal melanocytes, dermal stem cells (DSCs), and hair follicle stem cells.RESULTS:Junctional nevi and compound nevi were prone to occur on glabrous skin, such as the palms, soles, and vulva, and on the extremities in children, whereas intradermal nevi tended to develop on the trunk, head, and face of adults. The immunohistochemical data revealed that both junctional nevi and intradermal nevi expressed the epidermal melanocyte surface markers E-cadherin, DDR-1, and integrin α6 and the DSC molecular markers NGFRp-75 and nestin. CD34 was expressed only in junctional nevi, whereas K19 was not expressed in any type of melanocytic nevi. There was no significant difference in molecular expression at different sites or in different ages of onset. Nestin expression was markedly stronger in the intradermal nevi than in the junctional nevi, but there was no difference between the superficial and deep nevus cell nests of intradermal nevi.CONCLUSION:AMN may have a multicellular origin that commonly follows the mode of Abtropfung. Furthermore, DSCs may partly or independently participate in the formation of nevus cells.

黑素细胞痣(melanocytic nevus)又名色素痣,是皮肤的黑素细胞良性增生性病变,可生长于身体的任何部位,且以颜面部多发[1].因现代人们对外貌的要求并担心其恶变,色素痣手术切除逐年增加,成为病理科较常见的组织标本之一.根据痣细胞巢的生长部位可分为交界痣、复合痣、皮内痣.色素痣组织切片经HE染色后可见形状不一的棕黄色、棕褐色或棕黑色的细小颗粒,我们称之为黑色素.这种黑色素颗粒可掩盖细胞的结构和形态,影响观察,而且其颗粒存积的颜色与进一步免疫组化DAB显色的棕黄色难以区别,对疾病的诊断造成障碍.为去除黑色素颗粒对细胞的影响,提高色素痣标本的制片质量,笔者对传统脱黑色素法进行了探索改良,并对脱色素标本免疫组化的处理方法条件优化,现将实验内容介绍如下.

Microcystic/reticular schwannoma is a recently described, rare, distinctive histological variant of schwannoma with a predilection for the gastrointestinal tract [1]. Unlike classic Schwannoma, most microcystic/reticular Schwannoma had an incomplete fibrous capsule, two distinctly different areas - hypercellular areas and myxoid areas, Microcystic/reticular schwannoma occurs in the internal organs, especially in the digestive tract and a rare case occurs in the adrenal glands [1–3]. Herein, we report a case of microcystic/reticular schwannoma of the adrenal gland, in a 60-year-old male with 7.0 cm × 6.0 cm × 5 cm mass in the left adrenal gland. The tumor showed a vague multinodular appearance with a pushing border and arranged in a microcystic and reticular growth pattern with anastomosing and intersecting strands of spindle cells in a myxoid or collagenous/hyalinized stroma. Tumor cells showed diffuse nuclear and cytoplasmic positivity for S-100.

Extragastrointestinal stromal tumors (EGISTs) are rare tumors that arise outside the digestive tract. We report a case of an EGIST arising in the subcutaneous tissue of the abdominal wall, which at this site can often be misdiagnosed as dermatofibrosarcoma protuberans. The tumor was surgically resected from a 72-year-old male Chinese Han patient, and pathological examination revealed spindle-shaped tumor cells with eosinophilic cytoplasm and an oval nucleus. Immunohistochemically, the tumor cells showed strong cytoplasmic positivity for CD34, c-KIT (CD117), and DOG1. Tests for activating mutations of GISTs showed that the tumor cells carried an in-frame deletion (NP_000213.1:p.Lys550_Gln556del) in exon 11 of c-KIT (CD117). Thus, an EGIST should be considered in patients with abdominal subcutaneous tumors with an epithelioid, spindle-shaped, or mixed morphology. Immunohistochemistry of c-KIT (CD117) and DOG1 and genetic testing for activating mutations are recommended to aid in the differential diagnosis of subcutaneous tumors. In short, although EGISTs are rare in the abdominal subcutaneous tissue, pathologists must be aware of their possibility.

Triple-negative breast cancer (TNBC) is a complex heterogeneous disease that lacks the expressions of hormone receptors (HR) and human epidermal growth factor receptor 2 (HER2). Although TNBC make up less than 20% of breast cancer, it accounts for a large number of metastatic cases and deaths. Currently, extensive efforts have been made to look for potentially biomolecular targets for TNBC treatment. Based on the differences of molecular events identified by gene profiling analysis, the TNBC may be divided into some broad categories: basal-like (BL), mesenchymal-like (ML), immune-associated, HER2-enriched and luminal/apocrine breast cancers. Most of these are in the BL-TNBC category. BL-TNBC carries a representative molecular event of DNA-repair deficiency that increases the effectiveness of DNA destabilizers (represented by platinum agents) and DNA-damage response inhibitors (represented by PARP inhibitors). However, the results from clinical and preclinical studies have been inconsistent. Herein, we simply outline the progress of breast cancer classification and the significance of DNA repair deficiency in the clinic treatment for TNBCs. Previous studies have shown that the neoadjuvant therapies with platinum agents are effective for early-stage and metastatic TNBC patients with DNA repair defects. The results indicate that proper biomarkers (such as homologous recombination repair defects, HRD) are necessary for predicting the response to chemotherapy and often sufficient to select patients in early-phase treatment. Furthermore, the combination of chemotherapy drugs and inhibitors of DNA damage response represents a potential therapeutic strategy for TNBCs.

显微镜下结肠炎( microscopic colitis, MC)是一类慢性水样腹泻,内镜下结肠黏膜正常或轻度异常,病理组织活检呈特异性改变的疾病。近年来该病发病率逐年上升,约占慢性腹泻的13%[1],越来越受到临床医师的关注。 MC肠镜下黏膜并无特殊表现,而其他的相关影像学及一般的实验室检查对该病的诊断并无帮助,因此结肠镜组织活检显得尤为重要[2],而对活检组织进行 Masson特殊染色合并 CD3、CD45免疫组化染色对该病的诊断起到非常关键的作用,现将实验方法介绍如下。

对肌肉活检标本进行还原型辅酶Ⅰ四氮唑还原酶(NADH-tetrazolium reductase, NADH-TR)染色可区分不同的肌纤维结构,是神经肌纤维病变诊断的重要指标之一[1],免疫组化染色有利于观察肌纤维不同部位蛋白变化而进一步确定病变性质[2].在同一张切片上进行NADH-TR染色与免疫组化双重染色,可同时显示不同染色成分,为病理诊断提供更丰富的形态学依据.现将方法介绍如下.