奶牛乳腺上皮细胞和成纤维细胞的体外培养与鉴定

为体外培养纯化出稳定的奶牛乳腺上皮细胞和成纤维细胞,试验通过外科手术的方法取妊娠后期或泌乳期的荷斯坦奶牛乳腺组织,分离乳腺腺泡,用组织块法体外培养奶牛乳腺细胞,应用差时胰酶消化法和差速贴壁法将奶牛乳腺上皮细胞和成纤维细胞分别纯化出来,并用免疫组化的方法对细胞的纯度进行鉴定。结果表明:纯化的奶牛乳腺上皮细胞多为多角形,细胞核呈圆形或椭圆形,核仁清晰可见,多呈鹅卵石样或铺路石样生长,并可分泌乳滴,角蛋白-18反应阳性,波形蛋白反应阴性;纯化的奶牛乳腺成纤维细胞多为长梭形,呈旋涡状或放射状生长,角蛋白-18反应阴性,波形蛋白反应阳性;经纯化后2种细胞的纯度均可达95%以上,可满足后续试验的要求。     

Acute megakaryoblastic leukemia with erythrophagocytosis and thrombosis in a dog

A 7-year-old, intact male Dachshund was presented to the Lyon veterinary school for lethargy and anorexia of several weeks duration. The main clinical signs were pale and icteric mucous membranes, hepatomegaly, splenomegaly, and lymphadenopathy. Results of a CBC and plasma biochemistry tests revealed severe nonregenerative anemia, thrombocytopenia, and increased alanine aminotransferase and alkaline phosphatase activities. Blood smear evaluation and cytologic examination of lymph node and bone marrow aspirate specimens revealed a large population of poorly differentiated blast cells with morphologic features suggesting megakaryocytic lineage. A low number of well-differentiated but dysplastic megakaryocytes also were observed in lymph node and bone marrow smears. A few blast cells were erythrophagocytic. Blast cells were positive for glycoprotein IIIa, factor VIII-related antigen, and factor XIII using immunocytochemistry. The dog was euthanized and necropsied. Histologic findings consisted of diffuse, massive infiltration of lymph nodes, liver, and spleen by megakaryoblasts and atypical megakaryocytes, with widespread thrombosis. This case confirms the usefulness of immunochemistry, including for factor XIII, in the diagnosis of megakaryoblastic leukemia, and demonstrates the unique features of tumor cell erythrophagocytosis and marked fibrinous thrombosis, which have not been reported previously in dogs.     

Cytologic and histologic features of a poorly differentiated glioma in a dog

A 5-year old female Boxer with a 1-week history of progressive paresis and paraplegia had a T10-13 subarachnoid filling defect on myelography. Exploratory hemilaminectomy revealed an intramedullary spinal cord tumor which was subsequently diagnosed as a poorly differentiated glioma, most likely an anaplastic ependymoma. The cytologic, histologic, and immunocytochemical staining characteristics of this neoplasm are described. Differential diagnoses, including primary and secondary tumors involving the central nervous system are discussed.     

Malignant histiocytosis in a domestic cat: cytomorphologic and immunohistochemical features

Malignant histiocytosis (MH) was diagnosed in a 13-year-old neutered male Domestic Shorthair cat on the basis of light microscopic and immunohistochemical findings. Thoracic fluid analysis showed a modified transudate which contained a very few atypical discrete cells. Cytologic and histologic evaluation of mediastinal and splenic masses revealed a pleomorphic population of large, discrete, round cells 10 to 30 micrometers in diameter with marked cellular atypia. Nuclei were oval to reniform, often with prominent, bizarre nucleoli. Multinucleated cells and mitotic figures were commonly seen. Erythro- and leucocytophagia were noted. Immunohistochemistry indicated a scattered positive staining pattern with the histiocytic antigenic marker Mac387 and a minor population of cells showing positive reactivity for lysozyme. This report describes the characterization of MH in a cat and emphasizes that MH should be considered as a differential diagnosis in proliferative disorders of discrete-cells in this species.     

Development of a new canine osteosarcoma cell line

Establishing a canine osteosarcoma (OSA) cell line can be useful to develop in vivo and in vitro models of OSA. The goal of this study was to develop, characterize and authenticate a new canine OSA cell line and a clone. A cell line and a clone were developed with standard cell culture techniques from a naturally occurring OSA in a dog. The clonal cell line induced a tumour after injection in RAG 1‐deficient mouse. Histology was consistent with OSA. The original tumour from the dog and the tumour induced in the mouse were both reactive with vimentin and osteonectin (ON). The parent cell line and clonal cell line were reactive with ON, osteocalcin and alkaline phosphatase. Loss of heterozygosity was found in the same three microsatellite markers in the parent and clonal cell lines, and the tumour tissue grown in the mouse.     

Two cases of cutaneous T cell lymphoma in Friesian cows in the Azores

This paper presents two cases of cutaneous neoplasia in Friesian cows, on the island of S. Miguel, Azores. The lesions consisted of raised pink plaques, with no pruritus or signs of associated pain, that were extensively distributed over both lateral and ventral body surfaces. The histological examination of the material obtained by skin biopsy or post-mortem suggested a T cell lymphoma with extensive epidermal infiltration. The lymph nodes showed extensive invasion by tumour cells. The results of immunocytochemistry, which were the same for the tumour cells in the skin and in the lymph node, were positive for CD3, confirming the T cell origin of the tumour cells. It was concluded that these were cases of T cell lymphoma, involving both skin and lymph nodes. Although there have been other reports of cutaneous lymphosarcoma in the literature, these are the first cases identified in the Azores.     

Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats

Epithelial–mesenchymal transition (EMT) plays a crucial role in metastasis of epithelial tumors; however, it is challenging to detect EMT by cytology. In the present study, EMT was visualized by fluorescence-immunocytochemistry (FICC). Air-dried smears from epithelial tumors of dogs (n=22) and cats (n=9) were stained using mouse monoclonal anti-E-cadherin and rabbit monoclonal anti‐vimentin antibodies. Enzymatic immunohistochemistry (IHC) revealed that 51.6% (8/22 in dogs, 8/9 in cats) of the cases showed EMT. In dogs, FICC could detect EMT in 62.5% (5/8) of those cases. In cats, FICC could detect EMT in 100% (8/8) of the cases. In conclusion, the present FICC method could successfully detect EMT using conventional air-dried cytology smear slides.     

3种性类固醇激素及其受体在北方山溪鲵卵母细胞发育中的分布

为探讨性类固醇激素对北方山溪鲵(B a trachup erus tibetanus)卵泡发育的调控作用,采用组织学与免疫细胞化学方法,观测卵子发生周期中雌二醇(17-βestrad io l,E2)、孕酮(progesterone,P)、睾酮(testosterone,T)3种性类固醇激素及其受体(雌激素受体(E strogen R eceptor,ER)、孕激素受体(P rogesterone R eceptor,PR)、雄激素受体(A ndrogen R eceptor,AR))在不同发育时期卵泡中的表达特征。结果可见,北方山溪鲵卵巢中Ⅰ期卵泡数量在10月达到最大值,Ⅱ期卵泡数量在翌年5月达到最大值,Ⅲ期卵泡数量在6月最多,Ⅳ期卵泡数量在11月最多。E2与ER在Ⅲ期卵泡卵母细胞胞质中的阳性反应最强,P与PR,T与AR均在Ⅳ期卵泡卵母细胞中的阳性反应最强。结果表明,北方山溪鲵的卵泡为异步发育,其从发生至成熟后排出需要的最短时间应为20~22个月。E2在卵母细胞的卵黄形成中起作用,P与T均对卵母细胞成熟起重要作用。3种性类固醇激素分泌的变化规律与北方山溪鲵卵泡发生、发育、成熟和排出的周期一致。