Erythroblastic sarcoma is a rare and aggressive hematologic malignancy presenting as a mass-forming extramedullary proliferation of immature erythroid cells. Myeloid/lymphoid neoplasms with JAK2 rearrangement, most often PCM1::JAK2, may show eosinophilia, myelofibrosis, and expansion of immature erythroid precursors. NUTM1 rearrangements, initially recognized as defining alterations of NUT carcinoma, have subsequently been identified in selected hematologic malignancies but remain exceptionally rare in myeloid neoplasms. Here, we report an adult erythroblastic sarcoma harbouring concurrent PCM1::JAK2 and a novel NOP10::NUTM1 fusion, accompanied by partial chromosome 8p deletion and complicated by secondary hemophagocytic lymphohistiocytosis. To our knowledge, this is the first reported adult erythroblastic sarcoma with concurrent PCM1::JAK2 and NOP10::NUTM1 fusions. This case extends the molecular spectrum of erythroblastic sarcoma and underscores the value of integrated morphologic, immunophenotypic, cytogenetic, and genomic assessment in diagnostically challenging erythroid neoplasms.

We report a case of a 30-year-old male patient with a germline heterozygous TET2 R1354fs mutation, who successively developed three distinct lymphoid malignancies: mixed cellularity classical Hodgkin lymphoma (MCCHL), angioimmunoblastic T-cell lymphoma (AITL), and T-cell acute lymphoblastic leukemia (T-ALL). Sequencing analysis of the oral mucosa and bone marrow specimens during the remission phase confirmed the germline origin of the TET2 mutation. This represents the first documented case of the sequential transformation across three distinct lymphoid malignancies attributable to a germline TET2 mutation, underscoring its pivotal role in lymphomagenesis and clonal evolution.

Parathyroid carcinoma (PC) is a rare and aggressive endocrine malignancy with limited treatment options. Current treatments such as chemotherapy and radiotherapy have demonstrated limited efficacy. Here, we report the case of a male patient who presented with symptoms including polydipsia, polyuria, and joint pain. Further examination revealed a neck lump, hypercalcemia, and hyperparathyroidism, leading to a diagnosis of PC after en bloc surgery. Seven months later, the patient developed local recurrence and lung metastases, which were resected via left lateral neck dissection and thoracoscopic wedge resection. A 422-gene panel test revealed the presence of epidermal growth factor receptor ( EGFR) p.L858R (c. T2573G) mutation, which may sensitize the EGFR-tyrosine kinase inhibitor response, and phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha ( PIK3CA) p.E545KV (c. G1633A) mutation. After multidisciplinary treatment discussions, the patient was treated with the multi-target tyrosine kinase inhibitor, anlotinib, resulting in survival benefits for 19 months. This case highlights the potential of targeted therapy in terms of long-term survival in patients with distant metastatic PC, as well as the importance of precision therapy guided by genome sequencing to identify potential therapeutic targets.

Epstein–Barr virus (EBV)-associated lymphoproliferative diseases (EBV-LPDs) are common complications that occur after solid organ transplantation or allogeneic hematopoietic stem-cell transplantation (HSCT). However, their occurrence and treatment post-chimeric antigen receptor-modified T (CAR-T) cell therapy has not been reported. Two patients had been diagnosed with EBV-positive aggressive B-cell lymphoma and experienced relapses after multiple lines of treatment. After receiving CAR-T cell therapy in tandem with autologous HSCT, the patients achieved complete remission. However, with a median time of 38.5 months after CAR-T cell therapy, B-cell-derived EBV-LPDs were diagnosed, and they were relieved through the administration of immune checkpoint inhibitor or B-cell-depleting agents. Collectively, our report suggests that EBV-LPDs may represent a long-term adverse event after CAR-T cell therapy, especially in patients who previously had EBV-positive disorders, and they can be resolved by immune normalization strategy or B-cell depleting therapy.

Introduction: Standardization of methods for the immunohistochemical detection of CD30 expression and improvement of its reproducibility is critical for determining patient eligibility for anti-CD30 therapy. The protocol of the VENTANA CD30 assay has been well-optimized based on a pilot study. The current study aims to evaluate the staining concordance of CD30 detected by nine alternative immunohistochemical (IHC) assays in addition to the VENTANA CD30 assay. This evaluation is pivotal for exploring more equivalent detection methods to assess CD30 expression in patients with malignant lymphomas. Methods: This multi-centre retrospective study aimed to enroll 700-1,000 patients diagnosed with malignant lymphomas from 17 institutes across China. Tissue sections were stained using the VENTANA CD30 assay or at least one of the nine IHC assays utilizing different antibodies (umAB256 (zsbio); Ber-H2 (Maixin); Ber-H2 (DAKO); JCM182 (Leica) and platforms (Ventana BenchMark; DAKO; Leica). The staining results were evaluated by a panel of three independent expert hematopathologists in China based on consensus criteria. The primary objective of the study was to evaluate the concordance of semiquantitative results of CD30 staining detected by nine IHC assays and the VENTANA CD30 assay. Consistency was assessed by the intraclass correlation coefficient (ICC). To observe the effect of CD30 expression level on the concordance of different assays, cases with high-level expression (such as cHL and ALCL) and with low-level expression (with positivity <10% cells) were included in the subgroup analysis. Results: In this interim analysis, 346 patients from 12 institutes were enrolled, and 245 of whom met the inclusion criteria, after excluding those who met the exclusion and rejection criteria. With the exception of Ber-H2 (DAKO) + Ventana BenchMark assay, which showed a moderate concordance with the VENTANA CD30 assay, the concordance between the other eight CD30 IHC assays and the VENTANA CD30 assay was good or excellent, regardless whether cases with low-level expression were included. Excluding cases with high-level expression, the concordance between Ber-H2 (DAKO) + Ventana BenchMark assay and the VENTANA CD30 assay remained moderate, the same result got in Ber-H2(Maixin)+Ventana BenchMark. When excluding cases with low-level expression, the concordance of the umAB256 (zhongshanjinqiao) + Ventana BenchMark and Ber-H2 (Maixin) + Dako with the VENTANA CD30 assay turned to be moderate similarly. Conclusions: Overall, the concordance of nine CD30 IHC assays with the VENTANA CD30 assay varied from moderate to excellent, suggesting the reliability of the application of these approaches. Additional efforts are needed to optimize those assays with moderate concordance to inform treatment decisions better.

Age-related clonal hematopoiesis is defined as the presence of somatic mutations of genes known to be recurrently mutated in hematologic malignancies in the peripheral blood of healthy individuals.1 When clonal hematopoiesis is associated with a mutation at a variant allele frequency of 0.02 or greater,it is termed"clonal hematopoiesis of indeterminate potential"(CHIP).

Parathyroid carcinoma (PC) is a rare and aggressive endocrine malignancy with limited treatment options. Current treatments such as chemotherapy and radiotherapy have demonstrated limited efficacy. Here, we report the case of a male patient who presented with symptoms including polydipsia, polyuria, and joint pain. Further examination revealed a neck lump, hypercalcemia, and hyperparathyroidism, leading to a diagnosis of PC after en bloc surgery. Seven months later, the patient developed local recurrence and lung metastases, which were resected via left lateral neck dissection and thoracoscopic wedge resection. A 422-gene panel test revealed the presence of epidermal growth factor receptor ( EGFR ) p.L858R (c. T2573G) mutation, which may sensitize the EGFR-tyrosine kinase inhibitor response, and phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha ( PIK3CA ) p.E545KV (c. G1633A) mutation. After multidisciplinary treatment discussions, the patient was treated with the multi-target tyrosine kinase inhibitor, anlotinib, resulting in survival benefits for 19 months. This case highlights the potential of targeted therapy in terms of long-term survival in patients with distant metastatic PC, as well as the importance of precision therapy guided by genome sequencing to identify potential therapeutic targets. Keywords Case report , parathyroid carcinoma , hyperparathyroidism , epidermal growth factor receptor , tyrosine kinase inhibitor , lung metastasis

Abnormalities of FGFR1 have been reported in multiple malignancies, suggesting FGFR1 as a potential target for precision treatment, but drug resistance remains a formidable obstacle. In this study, we explored whether FGFR1 acted a therapeutic target in human T-cell acute lymphoblastic leukemia (T-ALL) and the molecular mechanisms underlying T-ALL cell resistance to FGFR1 inhibitors. We showed that FGFR1 was significantly upregulated in human T-ALL and inversely correlated with the prognosis of patients. Knockdown of FGFR1 suppressed T-ALL growth and progression both in vitro and in vivo. However, the T-ALL cells were resistant to FGFR1 inhibitors AZD4547 and PD-166866 even though FGFR1 signaling was specifically inhibited in the early stage. Mechanistically, we found that FGFR1 inhibitors markedly increased the expression of ATF4, which was a major initiator for T-ALL resistance to FGFR1 inhibitors. We further revealed that FGFR1 inhibitors induced expression of ATF4 through enhancing chromatin accessibility combined with translational activation via the GCN2-eIF2α pathway. Subsequently, ATF4 remodeled the amino acid metabolism by stimulating the expression of multiple metabolic genes ASNS, ASS1, PHGDH and SLC1A5, maintaining the activation of mTORC1, which contributed to the drug resistance in T-ALL cells. Targeting FGFR1 and mTOR exhibited synergistically anti-leukemic efficacy. These results reveal that FGFR1 is a potential therapeutic target in human T-ALL, and ATF4-mediated amino acid metabolic reprogramming contributes to the FGFR1 inhibitor resistance. Synergistically inhibiting FGFR1 and mTOR can overcome this obstacle in T-ALL therapy.

目的： 探讨成人胃孤立性朗格汉斯细胞组织细胞增生症（LCH）的临床表现、病理特征、免疫表型、分子病理、鉴别诊断及预后。 方法： 分析3例胃孤立性LCH的临床病理特点、免疫表型及分子病理等，并总结相关文献。 结果： 例1男，23岁，胃镜发现胃窦体交界前壁有一息肉样隆起；例2男，22岁，胃镜发现胃体小弯侧有一隆起糜烂灶；例3男，46岁，胃镜发现胃体上段有一充血糜烂灶、中段有一息肉。3例镜下均见异常增生的朗格汉斯细胞，细胞中等大小，胞质略嗜酸性，核卵圆形，可见核沟，背景里有大量炎性细胞浸润，免疫组织化学CD1α、Langerin、S‐100蛋白均为阳性。例2、3检测出BRAF V600E基因突变。3例均行胸腹部CT或正电子发射计算机断层扫描排除其他系统LCH。 结论： 胃孤立性LCH非常罕见，预后良好，经验不足的医师对本病的诊断要点往往了解不充分，应注意与低分化腺癌鉴别，避免误诊为恶性肿瘤。.

Abstract Background Abnormalities of FGFR1 have been reported in multiple malignancies, which proposes FGFR1 as a potential target for precision treatment, whereas drug resistance remains a formidable obstacle simultaneously. Methods RNA-seq analysis, mouse tumor models, and flow cytometry were performed to identify that the FGFR1 was a potential target in T-ALL. RNA-seq, ATAC-seq, targeted metabolomics analysis, surface sensing of translation (SUnSET) assay, western blot assays, and qRT-PCR were used to investigate the underlying mechanisms of FGFR1-targeting resistance. Drug screening was conducted to identify a drug combination strategy for overcoming this drug resistance. Results We identified that FGFR1 was observably upregulated in T-ALL and inversely correlated with the prognosis of patients. Functional studies showed that the knockdown of FGFR1 suppressed T-ALL cells growth and progression both in vitro and in vivo. Whereas the human T-ALL cells were resistant to FGFR1 inhibitors. Mechanistically, we identified that ATF4 was markedly upregulated and was a major initiator for T-ALL resistance to FGFR1 inhibitors. Expression of ATF4 was induced by FGFR1 inhibitors through enhancing chromatin accessibility transcriptionally combined with activating translation via the GCN2-eIF2α pathway. Then, ATF4 remodeled the amino acid metabolism by stimulating the expression of multiple metabolic genes, and further maintained the activation of mTORC1, which contributed to the drug resistance of malignancies. Moreover, targeting FGFR1 and mTOR exhibited synergistically antileukemic efficacy. Conclusion These findings revealed that FGFR1 was a potential therapeutic target in T-ALL, whereas ATF4-induced amino acid metabolic reprogramming mediated the FGFR1-targeted resistance. Synergistically targeting FGFR1 and mTOR could overcome this obstacle in T-ALL therapy.

Aims: We aim to evaluate the expression of GNA13 in germinal center (GC)-derived B cell lymphomas to evaluate its potential use as marker for diagnosis. Methods and Results: In this study, GNA13 expression was detected by immunohistochemistry in 129 cases of B-cell non-Hodgkin lymphomas (B-NHLs) and 8 cases of angioimmunoblastic T-cell lymphoma (AITL). Furthermore, mutations in the GNA13 gene in 114 cases of B-NHLs were detected by exome sequencing. In 50 cases of diffuse large B-cell lymphoma (DLBCL), 11 cases were positive for GNA13, and all GNA13-positive cases were of the germinal center B-cell-like (GCB) subtype. In 42 cases of follicular lymphoma (FL), the GNA13-positive rate was 100% in FL grade 1, 100% in FL grade 2, 80.0% in FL grade 3 A, and 63.2% in FL grade 3B. In 6 cases of Burkitt lymphoma, 3 cases were positive for GNA13. However, in small lymphocytic lymphoma, mantle cell lymphoma and marginal zone lymphoma, GNA13 was negative or only expressed in the GC. In addition, the GNA13-positive rate was 100% in AITL. The mutation of the GNA13 gene was 27.8% (5/18) in GCB-DLBCL and 10.3% (3/29) in FL, respectively, and mutation of GNA13 occurred in cases with weak positive or negative expression of the GNA13 protein. Conclusion: GNA13 may be a reliable new marker for GC cells applied in the diagnosis of GC-derived B-cell lymphomas. Its deficiency or low expression may be related to the degree of invasion of GC-derived B-cell lymphomas, and may result from genetic mutations.

OBJECTIVE:To investigate the expression and significance of insulinoma associated protein 1 (INSM1) and SRY-related high-mobility group box 11 (SOX11) in pancreatic neuroendocrine tumor (PNET) and solid pseudopapillary neoplasm (SPN). METHODS:To detect the expression of INSM1, SOX11, Syn, CgA, CD56, β-catenin, and CD99 in 56 cases of PNET, 42 cases of SPN, 16 cases of ductal adenocarcinoma (DACC) and 8 cases of acinar cell carcinoma (ACC) by immunohistochemistry. The application value of combination of INSM1 and SOX11 was compared with conventional markers (Syn, CgA, CD56, β-catenin, and CD99) in diagnosis and differential diagnosis of PNET and SPN. RESULTS:(1) In the 56 cases of PNET, the positive signals of INSM1 were located in the tumor and islet nucleus, the positive expression rate in the tumor tissues was 91.07% (51/56), whereas the signal was absent in 42 cases of SPN, 16 cases of DACC and 8 cases of ACC, and there were significant statistical difference between PNET with SPN, DACC, and ACC respectively (P < 0.001). (2) The positive signals of SOX11 were located in the tumor nucleus, with the positive expression rate was 92.86% (39/42) in SPN, however, the positive expression rate of SOX11 was 8.93% (5/56) in PNET, which included 3 cases of G1 and 2 cases of G3 types of PNET, the SOX11 positive signal was absent in 16 cases of DACC, 8 cases of ACC and peritumoral nomal pancreatic tissue, and the differences were statistically significant of positive rate between SPN with PNET, DACC and ACC, respectively (P < 0.001). (3) The sensitivity of INSM1(+)/SOX11(-) immunophenotype for PNET was 85.71%, vs. CD56 (57.14%), the difference was statistically significant (P=0.001); vs. Syn (80.36%) and CgA (71.43%), the difference was no statistically significant (P>0.05). The specificity of INSM1(+)/SOX11(-) for PNET was 100.00%, vs. Syn (42.86%) and CD56 (47.62%), the difference was statistically significant (P < 0.001); vs. CgA (92.86%), the difference was no statistically significant (P>0.05). The sensitivity of INSM1(-)/SOX11(+) immunophenotype for SPN was 92.86%, vs. β-catenin (90.48%) and CD99 (85.71%), the difference was no statistically significant (P>0.05). The specificity of INSM1(-)/SOX11(+) for SPN was 96.43%, vs. CD99 (48.21%), the difference was statistically significant (P < 0.001); vs. β-catenin (100.00%), the difference was no statistically significant (P>0.05). (4) The positive expression of INSM1 and SOX11 in PNET and SOX11 were not correlated with clinicopathological parameters (age, gender, tumor size, location, grade, and metastasis) (P>0.05). CONCLUSION:The positive expression patterns of INSM1 and SOX11 in PNET and SPN respectively are conductive to distinguish the both tumors. The combination of both take precedence over some corresponding conventional immunohistochemical markers in terms of sensitivity and specificity.

BackgroundAnaplastic Large Cell Lymphoma (ALCL) is one of the most common subtypes of T-cell lymphoma. Among these, refractory and relapsed (r/r) ALK positive ALCL lacks effective therapies. The chimeric antigen receptor-modified T (CAR-T) cell therapy holds great promise as a therapeutic strategy for this disease. However, it is not known yet whether anti-CD5 CAR-T cells are sufficient for the definitive treatment of relapsed ALK+ ALCL, nor the role of accurate laboratory-based diagnoses during CAR-T treatment.Case presentationThe adolescent patient received autologous T cells containing sequences encoding VH domains specific to CD5. Following the infusion, there was an increase in both the copy number and proportion of CAR-T cells in peripheral blood. IL-6 and ferritin levels in the patient exhibited significant fluctuations, with increases of 13 and 70 folds respectively, compared to baseline after the treatment. Additionally, adverse effects were observed, including grade 4 rash, grade 1 headache, nausea, and neck-pain. Surprisingly, a relapsed disease phenotype was identified based on the results of PET/CT and histopathological analysis of the inguinal lymph node biopsy. After conducting a thorough diagnostic assessment, which included flow cytometry, next-generation sequencing (NGS), examination of immune-related gene rearrangements, and analysis of the immune repertoire of T-cell receptors (TCR), we conclusively determined that the hyperplastic T cells identified in the lymph node were the result of an expansion of CAR-T cells. Ultimately, the patient has attained complete remission (CR) and has sustained a disease-free survival state for 815 days as of the cutoff date on August 30, 2023.ConclusionTaken together, the results demonstrate that anti-CD5 CAR-T cells can induce a clinical response in r/r ALK+ ALCL patient. Furthermore, this case underscores the importance of utilizing advanced technologies with high sensitivity and accuracy for biological detection in clinical laboratory diagnosis and prognosis in CAR-T cell treatment.Trial registration numberNCT04767308.

BackgroundMyeloid Sarcoma with monocytic differentiation is rare and quite likely is missed by surgical pathologists. However it is frequently misdiagnosed because of its non-specific imaging and histological pattern.Case presentationWe report the case of a 64-year-old woman with gastric primary myeloid sarcoma with monocytic differentiatio. Upper endoscopy revealed a neoplastic growth at the junction of the lesser curvature and gastric antrum. Except for a slightly increased peripheral monocyte count, no abnormalities were found on hematological and bone-marrow examination. Gastroscopic biopsy showed poorly differentiated atypical large cells with visible nucleoli and nuclear fission. Immunohistochemistry showed positive CD34, CD4, CD43, and CD56 expression, and weakly positive lysozyme expression. Immune markers for poorly differentiated adenocarcinoma, malignant melanoma, and lymphohematopoietic-system tumors were negative. The final diagnosis was myeloid sarcoma with monocytic differentiation. Chemotherapy did not shrink the tumor, so, radical surgery was performed. Although the tumor morphology did not change postoperatively, the immunophenotype did. CD68 and lysozyme expression (tumor tissue markers) changed from negative and weakly positive to strongly positive, AE1/3 expression (epithelial marker) changed from negative to positive, and CD34, CD4, CD43, and CD56 expression (common in naive hematopoietic cell-derived tumors) was greatly attenuated. Exome sequencing revealed missense mutations in FLT3 and PTPRB, which are associated with myeloid sarcoma, and in TP53, CD44, CD19, LTK, NOTCH2, and CNTN2, which are associated with lymphohematopoietic tumors and poorly differentiated cancers.ConclusionWe diagnosed myeloid sarcoma with monocytic differentiation after excluding poorly differentiated adenocarcinoma, common lymphohematopoietic-system tumors, epithelioid sarcoma, and malignant melanoma. We identified that the immunophenotypic of patient had alterations after chemotherapy, and FLT3 gene mutations. We hope that the above results will improve our understanding of this rare tumor.

Background: Despite of the paradigm change on the treatments of acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL) by venetoclax, it has been less successful in the treatment of diffuse large B-cell lymphoma (DLBCL). Here, we explored whether acylglycerol kinase regulates the sensitivity of DLBCLs to venetoclax and its mechanism in both cell lines and preclinical animal models. Methods: The expression of AGK and sensitivity to venetoclax of seven DLBCL cell lines were determined. Upon knockdown and overexpression of AGK by lentivirus in DLBCL cells, the venetoclax-induced apoptosis and PTEN-FOXO1-BCL-2 signaling axis were evaluated in vitro. The efficacy of venetoclax and PTEN-FOXO1-BCL-2 signaling axis were evaluated in immunodeficient NCG mice that were implanted with control or shAGK stably transduced SU-DHL4 cells. The expressions of AGK, BCL-2 and FOXO1 were evaluated in tumor tissues of DLBCL patients. Results: AGK expression was inversely correlated with sensitivity of DLBCL to venetoclax. Inhibition of AGK rendered the DLBCL cells more sensitive to venetoclax. Mechanistically, AGK phosphorylated and inactivated PTEN, which led to AKT activation and reduced FOXO1 nuclear translocation. Inhibition of AGK also led to enhanced efficacy of venetoclax for suppression of DLBCL tumor growth in vivo, which was dependent on FOXO1. In human DLBCL tumor tissues, the expression of AGK inversely correlated with BCL-2 expression, as well as the amounts of nuclear FOXO1. Conclusions: Our data demonstrated that AGK regulates venetoclax response in DLBCL via PTEN-FOXO1-BCL-2 signaling axis. Targeting AGK may enhance the efficacy of venetoclax for the treatment of DLBCL patients.

Objective: To investigate the clinical features, morphological characteristics, immunophenotype, and differential diagnosis of goblet cell adenocarcinoma (GCA) in the digestive system. Methods: The clinicopathological data, morphological characteristics, immunophenotypes of 22 cases of GCA in the digestive system diagnosed from January 2010 to January 2021 were collected. Meanwhile, 25 cases of neuroendocrine neoplasm (NEN) and 24 cases of adenocarcinoma were used as controls. Relevant literature was also reviewed. Results: There were 16 males and 6 females, aged from 36 to 79 years with an average of 56 years. The anatomical sites of the 22 GCA were mostly appendix (17 cases) and occasionally extra-appendix (5 cases), including 3 cases in stomach, 1 case in duodenum and 1 case in anal. All 17 cases of appendiceal GCA were pure GCA. Among the 5 cases of extra-appendiceal GCA, One case of gastric GCA was pure, two cases of gastric GCA with NEN or adenocarcinoma, duodenal GCA with NEN and adenocarcinoma, anal GCA with NEN.Low-grade GCAs were composed of goblet, Paneth and neuroendocrine cells, which were arranged in intestinal crypt tubular or cluster structures and distributed in the wall of digestive system. The tubular and cluster structures lacked adhesion. Goblet cells were columnar, located in the base, with clear cytoplasm, small nuclei, inconspicuous atypia, and uncommon mitoses. Extracellular mucus and signet-ring cells with nuclear variations could be seen in some cases. Nerve fiber bundle invasion and tumor thrombus in vessels were often present. High-grade GCAs lacked tubular and cluster structures, and their histological structures were more complex. Tumor cells expressed mixed neuroendocrine and glandular epithelial markers. Similar to the expression patterns of synaptophysin and chromogranin A, CD200 and INSM1 were also dot-like or patch-positive in GCA. Conclusions: GCA is an infrequent tumor of the digestive system and shows the bi-directional differentiation characteristics of neuroendocrine and glandular epithelium. Accurate diagnosis and staging are related to its prognosis.

目的 探讨胰腺实性-假乳头状肿瘤(solid-pseudopapillary neoplasms,SPN)的临床病理特征及转录因子E3(transcrip-tion factor E3,TFE3)表达在诊断中的价值.方法 采用免疫组化法检测32例胰腺SPN中TFE3、β-catenin、CD10、CK、vimen-tin、Syn、CD56、AR、PR的表达,40例胰腺非SPN分别为17例神经内分泌肿瘤(neuroendocrine tumor,NET)、15例导管腺癌(duct adenocarcinoma,DACC)、3例腺泡细胞癌(acinar cell carcinoma,ACC)、3例神经内分泌癌(neuroendocrine carcinoma,NEC)和2例胰母细胞瘤作为对照组,分析TFE3在SPN诊断和鉴别诊断中的应用价值.结果 β-catenin、CD10、CK、vimentin、Syn、CD56、AR、PR在胰腺SPN和非SPN中均呈不同程度表达.32例胰腺SPN中TFE3阳性位于肿瘤细胞核,其中28例呈弥漫强阳性,4例呈局部阳性,阳性率为100.0％(32/32);胰腺非SPN中有3例NET、1例NEC和1例ACC呈TFE3阳性,阳性率为12.5％(5/40);两组间差异有显著性(P<0.001).TFE3对胰腺SPN的敏感性和特异性分别为100.0％和87.5％.TFE3、β-catenin和CD10三者联合的敏感性和特异性分别为93.8％和100.0％,优于TFE3和TFE3联合β-catenin或CD10检测.结论 TFE3在肿瘤细胞核中呈强阳性表达模式是胰腺SPN独特的免疫表型特征;TFE3联合β-catenin、CD10检测具有高敏感性和高特异性,可以作为诊断胰腺SPN的重要诊断标志物.

目的 探讨非瑟酮(Fisetin)对阿霉素(Doxorubicin,DOX)诱导的人肺动脉内皮细胞(HPAEC)衰老的影响及其相关机制.方法 使用不同浓度的DOX(0.1、0.25、0.5、1μmol/L)诱导HPAEC衰老,通过CCK8法检测细胞增殖活性,Western blot法检测衰老相关蛋白P53、P21表达.随后用不同浓度的Fisetin作用于HPAEC,分别于0、24、48和72 h检测Fisetin对HPAEC细胞增殖的影响,流式细胞术检测Fisetin对细胞周期的影响,利用衰老相关β-半乳糖苷酶(SA-β-gal)对衰老细胞进行染色并计数,活性氧(ROS)试剂盒检测Fisetin对HPAEC活性氧(ROS)产生的影响,West-ern blot法检测衰老相关蛋白、Nrf2和MAPK信号蛋白的表达情况,酶联免疫吸附试验(ELISA)检测分泌的细胞因子浓度,CCK8法检测Fisetin预处理的HPAEC培养上清对人肺动脉平滑肌细胞(HPASMC)增殖的影响.结果 当Fisetin浓度在20μmol/L以下时,HPAEC增殖活性在24、48、72 h没有明显降低,差异无统计学意义(P>0.05);流式细胞术检测发现DOX刺激后,90％ 以上的HPAEC被阻滞在DNA合成前期(G0/G1期),而Fisetin(5、10、20μmol/L)处理组细胞周期在DNA合成期(S期)分别占40％、50％ 及55％,呈浓度依赖性上升,差异具有统计学意义(P<0.01).Western blot证实DOX刺激后,HPAEC中衰老相关蛋白P21表达增加,而随着Fisetin作用浓度的增加,P21表达逐渐下降(P<0.05).SA-β-gal衰老染色和ROS染色证实DOX干预后,衰老细胞的数量明显增加,ROS的产生量也增加;而Fisetin处理后,衰老细胞的数量逐渐减少,ROS的产生量也减少.Fisetin可增加HO-1、p-P38的表达,促进Nrf2核转移,沉默HO-1和抑制p-P38表达均可减弱Fisetin对P21的抑制作用.DOX刺激后细胞衰老相关分泌表型(SASP)的细胞因子表达不断升高,而不同浓度的Fisetin(5、10、20μmol/L)干预后,SASP细胞因子的表达量下降.CCK8实验证实,Fisetin预处理的HPAEC培养上清对HPASMC增殖有抑制作用.结论 Fisetin可通过Nrf2/HO-1和MAPK信号通路抑制DOX诱导的HPAEC衰老,并能够抑制HPASMC增殖,从而抑制肺动脉重塑.