骨外Ewing肉瘤/外周原始神经外胚叶肿瘤的临床病理分析

目的探讨骨外Ewing肉瘤／外周原始神经外胚叶肿瘤的临床病理特征及诊断、鉴别诊断依据。方法18例骨外Ewing肉瘤／外周原始神经外胚叶肿瘤行常规病理检查及免疫组化染色,其中2例进行电镜检查。结果光镜显示肿瘤组织主要由小圆形或卵圆形细胞组成,免疫组化染色显示肿瘤细胞膜CD99强阳性,电镜显示肿瘤细胞质内有神经内分泌颗粒。结论骨外Ewing肉瘤/外周原始神经外胚叶肿瘤的诊断依赖病理特征,并需要与其他小细胞恶性肿瘤进行鉴别。     

乳腺血管肉瘤临床病理分析

目的 分析乳腺血管肉瘤的临床及病理特点.方法 对2例乳腺血管肉瘤进行光镜及免疫组化染色观察.结果 乳腺血管肉瘤组织学形态呈多样性.免疫组化染色显示,血管肉瘤表达血管内皮标记物FⅧRAg、CD31、CD34及淋巴管内皮标记物D240.结论 乳腺血管肉瘤十分罕见,临床表现无特异性,病理形态多样,诊断困难,组织学形态结合免疫组化染色对于确诊有意义.     

上皮样血管肉瘤1例报道并文献复习

目的 探讨软组织上皮样血管肉瘤(epithelioid angiosarcoma,EAS)的临床病理特点、诊断和鉴别诊断.方法 报道1例臀部软组织EAS临床资料、光镜及免疫组化染色结果,并结合文献复习进行讨论.结果 光镜下肿瘤成分几乎全部为上皮样肿瘤细胞组成,细胞排列成小巢状、片状或条索状,可见原始血管结构形成.免疫组化染色:瘤细胞高表达CD34、CKpan及vimentin,并表达CD31、CD99;而SMA、S-100、Myoglohin及HMB-45均阴性;Ki-67增殖指数＜20%.结论 软组织血管肉瘤罕见但上皮样型更为罕见,经组织病理学详细观察及免疫组化染色协助,对于正确诊断和鉴别诊断有重要意义.     

卵巢转移性血管肉瘤1例报道并文献复习

目的 探讨卵巢转移性血管肉瘤的临床病理特点和鉴别诊断.方法 报道1例乳腺血管肉瘤卵巢转移的临床资料、光镜及免疫组化染色结果,结合文献讨论.结果 患者女性,78岁.右侧乳腺血管肉瘤术后10~24个月后发生卵巢肿块.临床表现无特异性,肿瘤边界清楚,局部卵巢结构被破坏,肿瘤由大小不一、不规则的腔隙性血管及实性细胞组成.血管腔隙衬覆的细胞呈立方或上皮样,实性细胞区见细胞呈圆形、椭圆形及梭形,异型性明显,核分裂象易见,CD31 和CD34均呈阳性.结论 卵巢转移性血管肉瘤较为少见,临床病史、免疫组化染色对诊断与鉴别诊断有重要意义.     

乳腺纤维母细胞/肌纤维母细胞性肿瘤临床病理分析

目的 探讨乳腺纤维母细胞/肌纤维母细胞性肿瘤(breast fibroblastic/ myofibroblastic tumors BF/MFT)的临床病理、免疫表型特点及相关鉴别诊断.方法 应用HE染色、免疫组化标记对7例BF/MFT进行形态学观察并进行文献复习.结果 7例患者均为女性,2例为孤立性纤维性肿瘤;2例为肌纤维母细胞瘤;1例为纤维瘤病;1例为炎性肌纤维母细胞瘤;1例为低度恶性肌纤维母细胞肉瘤.结论 BF/MFT非常少见,不同的类型肿瘤生物学行为不同,它们的鉴别诊断主要是组织学及免疫组化.     

伴神经内分泌分化的胃原发性癌肉瘤临床病理观察

目的 探讨伴神经内分泌分化的胃原发性癌肉瘤(gastric primary carcinosarcoma with neuroendocrine differentiation)的临床病理特征,诊断及鉴别诊断.方法 对1例伴神经内分泌分化的胃原发性癌肉瘤进行光镜观察、免疫组化标记和W-S染色,并复习相关文献.结果 患者因烧心、黑便40天就诊.胃镜发现胃贲门处有1巨大菜花状肿物.光镜下肿瘤主要由两种成分构成.一种为不规则腺管结构,免疫表型CK和EMA阳性,vimentin阴性,多数腺管Syn阳性,少数腺上皮细胞CgA阳性;另一种为弥漫分布的间质细胞,免疫表型为vimentin广泛阳性,actin、CK、EMA、Syn、S-100局灶性阳性,CgA阴性.瘤组织中可见局灶性软骨肉瘤、骨肉瘤及平滑肌肉瘤成分.肿瘤旁胃黏膜未查见幽门螺杆菌.结论 伴神经内分泌分化的胃原发性癌肉瘤是一种极其罕见,恶性程度较高,预后较差的肿瘤.其诊断与鉴别诊断主要依靠组织病理学及免疫组织化学.     

Askin瘤尸检1例并文献复习

目的 探讨Askin瘤的临床病理特点、诊断及鉴别诊断.方法 总结1例Askin瘤患者的临床资料和尸检病理学特点;通过光镜和免疫组化及网状纤维染色,对尸体解剖后进行组织病理学观察.结果 临床上表现为青少年胸肺部肿块伴疼痛,患者生存期短,预后差.病理组织学特征主要为肿瘤组织呈弥漫性或结节状分布.组织学上以稀少的纤维血管间质将形态单一的小圆、卵圆形瘤细胞围绕成巢状或小叶状结构.免疫组化显示:瘤细胞CD99(+),NSE弱(+),CgA弱(+).网状纤维染色示肿瘤细胞间网状纤维少.结论 Askin瘤为一种位于胸肺部神经内分泌源性的高度恶性肿瘤,它实际上是PNET/Ewing肉瘤家族的一员,只是发生部位较特殊,预后不良.     

弥漫性特发性肺神经内分泌细胞增生伴微小瘤形成1例并文献复习

目的 探讨弥漫性特发性肺神经内分泌细胞增生伴微小瘤形成的临床及病理特点.方法 对1例弥漫性特发性肺神经内分泌细胞增生伴微小瘤形成进行光镜和免疫组化检测并文献复习.结果 弥漫性特发性肺神经内分泌细胞增生及微小瘤常继发于肺间质性疾病,好发于中年人或非吸烟老年女性,临床表现为干咳和气短等气道堵塞症状,病情进展极其缓慢,体征可不明显.镜下表现为在支气管扩张、慢性炎细胞浸润、肺间质弥漫纤维化的基础上出现多灶性神经内分泌细胞增生(直径<5 mm),增生的细胞可局限于支气管和细支气管上皮或突破基膜向间质浸润.细胞呈短梭形或椭圆形,大小相对一致,核分裂象罕见.免疫组化染色Syn、CgA、CD56强阳性;广谱CK、CK-L、CK-H阳性;增殖指数Ki-67较低,p63、Vim等阴性.结论 弥漫性特发性肺神经内分泌细胞增生伴微小瘤形成是类癌的癌前病变,临床表现不明显,常为偶然发现,确诊主要靠病理学检查及免疫组化,临床处理以密切随访为主,必要时可行外科治疗,预后好.     

心包原发性恶性间皮瘤的临床病理分析

目的：探讨心包原发性恶性间皮瘤的临床病理特征。诊断与鉴别诊断要点。方法：对4例心包原发性恶性间皮瘤进行临床病理分析。光镜及免疫组化染色观察并复习有关文献。结果：男3例,女1例,平均年龄42岁,3例呈局限型,1例为弥漫浸润型。组织学上可表现为肉瘤样梭形细胞型,上皮样型及双相型,免疫组化染色显示肉瘤样梭形细胞表CK、vimentin;上皮样型瘤细胞表达HBME1、CK。结论：原发于心包的恶性间皮瘤罕见,预后极差。临床常被误诊,其组织形态亦复杂多样,应注意与心包的良性增生性病变,心包转移性腺癌和梭形细胞肿瘤等相鉴别。     

乳腺尤因肉瘤(ES)/原始神经外胚层肿瘤(PNET)1例报道并文献复习

目的 探讨乳腺尤因肉瘤/原始神经外胚层肿瘤(primitive neuroectodermal tumors,PNET)的病理诊断及鉴别诊断.方法 对1例乳腺尤因肉瘤/PNET进行免疫组化染色,并复习相关文献.结果镜下观察瘤组织由形态一致的小圆细胞所组成,呈实性片状或乳头状排列,局部可见菊形团样结构.免疫组化显示瘤细胞表达CD99、ER、CD56、GFAP、FLI-1、Calponin等.结论乳腺尤因肉瘤/PNET是少见的小圆细胞恶性肿瘤,CD99、FLI-1及神经内分泌标记物阳性对确诊有重要意义,ER可能在乳腺尤因肉瘤/PNET诊断中有一定的意义.     

Limitations in the ability of NB84 to detect metastatic neuroblastoma cells in bone marrow

BACKGROUND: The accurate assessment of metastases is an essential component of the staging process for children with neuroblastoma. AIMS: To study the sensitivity of the immunohistochemical marker neuroblastoma 84 (NB84) for the detection of bone marrow infiltrates in children with stage 4 neuroblastoma. METHODS: Primary tumour specimens, bone marrow trephine biopsy specimens and lymph node metastases, taken from children with neuroblastoma that had metastasised to bone marrow, were assessed with a panel of commonly used immunohistochemical markers for neuroblastoma. A comparison was drawn between the sensitivity of the marker NB84 for primary tumours and for bone marrow metastases. RESULTS: NB84 immunolabelled all pre-chemotherapy and post-chemotherapy (n = 24) paired primary tumour specimens, as well as each of a further 20, unpaired, pre-chemotherapy primary tumour specimens. It also labelled all (n = 4) lymph node metastases. Immunolabelling of bone marrow trephine biopsy specimens (21/33) was less sensitive. Of 16 primary tumour specimens with a paired bone marrow trephine biopsy specimen, all immunostained positive, whereas only 62.5% of bone marrow biopsy specimens immunostained positive for NB84. The number of bone marrow biopsy specimens immunostaining for NB84 was significantly lower than the number of paired primary tumour specimens (p = 0.041). CONCLUSIONS: NB84 remains a useful marker for the diagnosis of neuroblastoma in primary tumour specimens, but not for neuroblastoma that has metastasised to bone marrow.     

A murine model for bone marrow metastasis established by an i.v. injection of C-1300 neuroblastoma in A/J mice

A reproducible tumor model for bone marrow metastasis has been developed by an injection of murine C-1300 neuroblastoma (C-1300 NB) cells into the tail vein of syngeneic A/J mice. The animals died with liver metastases at 18–21 days after an injection of 10⁵ tumor cells and often had bone marrow metastasis in the femur. N-methylformamide (NMF), a maturational agent, was administered to inhibit liver metastases and to extend survival in mice with advancing bone metastasis. Histological examination of bone marrow metastasis, demonstrated lesions varying from a few small colonies of C-1300 NB cells either in metaphysis or diaphysis to large foci replacing normal hematopoietic bone marrow, simultaneously invading epiphysis or cortex of bone as bone metastasis. This assay demonstrated the ability to detect neuroblastoma cells in the bone marrow histologically and could determine bone marrow TD₅₀ by extraction of bone marrow cells after treatment with various doses of drug. Fifty per cent of mice injected with cyclophosphamide (CY) developed bone marrow metastasis without liver metastasis. Treatment with tamoxifen, an anti-calmodulin drug, suppressed tumor takes in the recipient mice with tamoxifen-dose-dependent fashion. This experimental system allows for investigations into the therapeutic response and biology of neuroblastoma metastases in the bone marrow.     

Immunohistochemical demonstration of neurone specific enolase in bone marrow infiltrated by neuroblastoma.

One hundred and two bone marrow samples were analysed by histological and immunohistochemical methods for neurone specific enolase (NSE). The biopsies were performed to determine the extent of bone marrow disease in 84 neuroblastomas, nine embryonal rhabdomyosarcomas, five Ewing's sarcomas, two cases of Hodgkin's disease and two lymphoblastic lymphomas. Twenty seven (32%) of neuroblastoma bone marrows showed metastases by conventional histological techniques and 33 (39%) after immunohistochemical staining with NSE. Five embryonal rhabdomyosarcomas, five Ewing's sarcomas, and two lymphoblastic lymphomas showed bone marrow metastases. Only one of these cases was reactive for NSE. NSE represents a very sensitive immunomarker for the follow up of neuroblastoma and improves detection of bone marrow invasion by neuroblastoma.     

Collaboration of cancer‐associated fibroblasts and tumour‐associated macrophages for neuroblastoma development

Neuroblastoma is the most common extracranial solid tumour in children and is histologically classified by its Schwannian stromal cells. Although having fewer Schwannian stromal cells is generally associated with more aggressive phenotypes, the exact roles of other stromal cells (mainly macrophages and fibroblasts) are unclear. Here, we examined 41 cases of neuroblastoma using immunohistochemistry for the tumour‐associated macrophage (TAM) markers CD68, CD163, and CD204, and a cancer‐associated fibroblast (CAF) marker, alpha smooth muscle actin (αSMA). Each case was assigned to low/high groups on the basis of the number of TAMs or three groups on the basis of the αSMA‐staining area for CAFs. Both the number of TAMs and the area of CAFs were significantly correlated with clinical stage, MYCN amplification, bone marrow metastasis, histological classification, histological type, and risk classification. Furthermore, TAM settled in the vicinity of the CAF area, suggesting their close interaction within the tumour microenvironment. We next determined the effects of conditioned medium of a neuroblastoma cell line (NBCM) on bone marrow‐derived mesenchymal stem cells (BM‐MSCs) and peripheral blood mononuclear cell (PBMC)‐derived macrophages in vitro. The TAM markers CD163 and CD204 were significantly up‐regulated in PBMC‐derived macrophages treated with NBCM. The expression of αSMA by BM‐MSCs was increased in NBCM‐treated cells. Co‐culturing with CAF‐like BM‐MSCs did not enhance the invasive ability but supported the proliferation of tumour cells, whereas tumour cells co‐cultured with TAM‐like macrophages had the opposite effect. Intriguingly, TAM‐like macrophages enhanced not only the invasive abilities of tumour cells and BM‐MSCs but also the proliferation of BM‐MSCs. CXCL2 secreted from TAM‐like macrophages plays an important role in tumour invasiveness. Taken together, these results indicate that PBMC‐derived macrophages and BM‐MSCs are recruited to a tumour site and activated into TAMs and CAFs, respectively, followed by the formation of favourable environments for neuroblastoma progression. © 2016 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.     

The significance of immunological analysis for the detection of residual neuroblasts in bone marrow

Immunological analysis is complementary to morphological investigation to detect bone marrow (BM) metastases in neuroblastoma patients. It is essential at time of BM harvest for an autograft, since it is the only examination which allows a precise evaluation of the graft contamination by malignant cells. Simple indirect immunofluorescence staining with anti-neuroblastoma monoclonal antibodies (UJ13A and HSAN 1-2) allows a final detection of about 1% malignant cells in the BM, and 1% when cells are gathered in clumps. The use of an immunocytochemical method (alkaline phosphatase) together with double immunofluorescence staining permits to detect as few as one residual neuroblastoma cell in 10(4) normal ones. These two methods used together have allowed to assess the neuroblastoma nature of rare isolated cells with pseudo-lymphocytic aspect.     

Bone marrow metastases: a survey of nonhematologic metastases with immunohistochemical study of metastatic carcinomas.

With current therapies and imaging methods, staging bone marrow biopsies, and subsequently found metastases to the bone marrow, are less frequent. Historically, the most common metastatic nonhematologic tumors to the bone marrow in adult and pediatric patients included breast carcinoma, neuroblastoma, prostatic carcinoma, Ewing sarcoma, and lung tumors. Although the staining patterns of these primary tumors have been extensively examined, the utility of these immunohistochemical profiles has not been studied in the context of metastatic lesions to the bone marrow. In our review of 111 metastases to the bone marrow over an 18-year period, the most common primary tumor types are, in order of frequency: breast carcinoma, neuroblastoma, lung tumors, rhabdomyosarcoma, Ewing sarcoma, prostate carcinoma, and gastrointestinal tract tumors. Additionally, in an analysis of 44 adult metastatic carcinomas, we confirm that immunohistochemical panels are useful in identifying the primary tumor site. Overall, the immunohistochemical characterization of metastatic carcinomas to the bone marrow show good correlation with the established staining pattern of the primary tumors.     

Histopathologic analysis of bone marrow and bone metastasis in murine neuroblastoma

To elucidate the development of bone metastasis in human neuroblastoma, bone marrow and bone metastases were analysed histologically in a hematogeneous metastasis model of murine neuroblastoma. The bone marrow metastasis occurred initially in the bone marrow sinusoid where tumor cells adhered and extravasated to bone marrow parenchyma, resulting in the formation of nodular lesions in the medullary cavity. The nodular lesions eventually progressed to diffuse lesions segmentally occupying the medullary cavity. During the establishment of the diffuse lesions, tumor cells invaded cancellous bone and/or bone cortex, resulting in bone metastasis. Such nodular or diffuse bone marrow metastatic lesions occurred sporadically in a variety of bones. To improve the results of treatment for neuroblastoma, the characteristics of the bone marrow and the bone metastases demonstrated in this study should be considered in the diagnosis and treatment of this disease.     

间变性淋巴瘤激酶在神经母细胞瘤的表达及其意义

目的探讨间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)在神经母细胞瘤(neuroblastoma,NB)中的表达及与预后的关系。方法应用免疫组化EnVision法检测NB内ALK的表达情况,应用国际NB病理分类及分期系统进行病理分类及分期,结合随访资料分析ALK的表达程度与预后之间的关系。结果 80例NB中ALK阳性70例,阳性率为87.5%,阳性表达定位于肿瘤细胞胞质及神经毡内,Schwannian纤维及间质中血管、淋巴细胞均阴性。ALK的表达程度在肿瘤的不同临床分期之间差异明显;患者的平均生存时间与ALK表达程度有关。结论 ALK在NB中异常表达,且其表达水平和NB的侵袭性及预后之间存在联系,即高表达间变性淋巴瘤激酶的肿瘤,更具有侵袭性,预后较差。     

Bone marrow metastatic myeloma cells promote osteoclastogenesis through RANKL on endothelial cells

We have been using the B9/BM1 murine bone marrow metastasis model to study the function of adhesion molecules in the cell-cell interactions and transendothelial migration, necessary for tumor metastasis. The cell surface phenotype of these cells, which colonize vertebral and femoral marrow after intravenous injection, shows great similarity to that of human myeloma cells. In the present study, we investigated the interaction between B9/BM1 cells and osteoclasts, which likely support tumor metastasis in bone marrow. We found that co-culturing B9/BM1 cells and bone marrow-derived endothelial cells (BMECs) in the presence of vitamin D₃ and M-CSF promoted differentiation of primary osteoclast progenitors to osteoclasts (detected by TRAP staining), and that this effect was blocked when BMECs were separated from the other cells by a porous polycarbonate membrane. Flow cytometry analysis showed that BMECs expressed RANKL (receptor activator of NF-κB ligand) protein on their surface, and that this expression was up-regulated by co-culture with B9/BM1 cells. Accordingly, RT-PCR showed expression of RANKL mRNA also to be up-regulated in BMECs co-cultured with B9/BM1 cells. Addition of OPG (osteoprotegerin, a decoy RANKL receptor) to the co-culture system completely blocked osteoclast induction, as did addition of anti-CD44 antibody. Furthermore, intravenous injection of B9/BM1 cells substantially increased the numbers of TRAP-positive osteoclasts detected in mice in vivo. Taken together, these findings suggest that B9/BM1 myeloma cells act via CD44 to stimulate RANKL expression on BMECs, which in turn physically interact with osteoclast progenitors to promote their differentiation to osteoclasts and metastasis in bone marrow. This revised version was published online in July 2006 with corrections to the Cover Date.     

Immunomagnetic depletion of malignant cells from autologous bone marrow graft: from experimental models to clinical trials

Using experimental modes with normal allogeneic bone marrow (BM) contaminated with Burkitt or neuroblastoma cell lines, and a liquid culture assay, we demonstrated that, when used in optimal conditions, the immunomagnetic depletion technique permitted a reproducible elimination of 3-4 log malignant cells. Results were very similar to those obtained with the complement lysis procedure in Burkitt lymphoma. This immunomagnetic procedure was used in 123 cases of autologous bone marrow transplantation (ABMT) in children with neuroblastoma. The analysis of the cases demonstrated, first, that the procedure induced a significant loss of mononuclear cells but was not toxic for BM precursors. Delays to engraftment observed in a few patients were probably due to the combination of pejorative factors, especially the damage caused to the micro-environment by previous heavy and prolonged chemotherapy or the double ABMT programme. Second, patients presented with profound T-cell defect with undetectable IL2 secretion up to 1 year post-graft but they all had normal NK functions from the first month post-graft, these functions exceeding normal values on the second and third months post-graft. Finally, in 20 cases, dual-immunofluorescence staining permitted the demonstration that the autograft contained malignant cells before purging that were eliminated by the immunomagnetic depletion.