小鼠出生后背部毛囊发育与Lef-1表达的研究

旨在研究Lef-1在小鼠背部毛囊生长周期中的表达规律及其与毛囊生长更替的关系。本试验采用石蜡切片HE染色、Western blotting以及免疫组织化学方法,研究小鼠毛囊的周期性变化,检测Lef-1在小鼠毛囊生长的不同时期皮肤组织中的蛋白表达水平,并对其进行组织定位。结果,昆明系小鼠毛囊生长呈周期性变化,处于不同生长时期的毛囊具有各期的特殊结构;Western blotting结果显示,在各阶段皮肤组织中存在Lef-1蛋白的表达,其在毛囊生长初期的表达量显著高于中期和末期;免疫组织化学结果显示,Lef-1在不同毛囊生长时期的皮脂腺、根鞘和真皮乳头均有不同程度的表达。Lef-1的表达与毛囊的周期性变化存在相关性。     

ALV-J诱发鸡成红细胞白血病的临床病例分析

2010—2011年检测3批病例,分别来自泰安新泰的110日龄麻鸡、济宁泗水的90日龄麻鸡和济南20日龄蛋雏鸡。病鸡均表现消瘦、精神萎靡、嗜睡、鸡冠稍苍白或发绀等,死亡率分别为20%、12%和18%。剖检,3批病鸡均可见肝脏、脾脏及肾脏严重肿大,胸腺、肌肉、腺胃等组织器官出血。新泰病鸡肝脏和肺脏表面有明显的白色肿瘤结节。血液涂片与骨髓涂片观察发现大量红细胞转变为体积较大,胞质丰富蓝染,胞核大呈圆形、核内有很纤细的染色质、核周围有空泡的成红细胞,以晚幼成红细胞为主;组织学检查发现,在所有组织脏器血管及间质内均可观察到大量聚集的与血涂片中形态相一致的成红细胞,并且可观察到核分裂相,但未见淋巴细胞聚集。通过血液学和组织学观察不仅排除马立克氏病和网状内皮组织增生症,而且排除中毒及代谢性疾病的可能。以肝脏组织DNA为模板,在3批病鸡中利用PCR检测均扩增出J亚群禽白血病病毒(ALV-J)gp85基因924bp的特异性片段,而A亚群禽白血病病毒(ALV-A)、B亚群禽白血病病毒(ALV-B)阴性。免疫组织化学检测结果表明,发病鸡肺脏和脾脏等含血量丰富的组织内发现组织细胞及成红细胞胞质呈ALV-J抗原阳性。根据以上检测结果确定此3批患有成红细胞白血病病鸡均由ALV-J感染引起。ALV-J引起单纯鸡成红细胞白血病在国内为首次发现。ALV-J在我国鸡群中的多潜能致瘤机制需要进一步的研究。     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

A retrospective study of the clinical, histological, and immunohistochemical manifestations of 5 dogs originally diagnosed histologically as necrotizing scleritis

Purpose To describe the clinical, histological, and immunohistochemical manifestations of canine necrotizing scleritis. Methods A retrospective examination of the clinical records and samples of ocular tissues from five dogs with a histological diagnosis ‘necrotizing scleritis’ was completed. Archived, formalin‐fixed, paraffin‐embedded samples and two control globes were stained with hematoxylin and eosin, Gram, periodic acid–Schiff (PAS) and Masson trichrome stains, and they were immunohistochemically labeled for CD3, CD18, and CD20. Results Of the five cases reviewed, only two could be confirmed as idiopathic necrotizing scleritis. The other three cases were retrospectively diagnosed as unilateral focal, non‐necrotizing scleritis, one as episcleritis and the third was scleritis secondary to a proptosed globe based on our retrospective clinical, histological, and immunohistochemical evaluations. In these two cases, idiopathic necrotizing scleritis manifested as a bilateral, progressive, inflammatory disease of the sclera and cornea that induces significant uveitis. Light microscopic examination confirmed collagen degeneration and granulomatous inflammation. There was no evidence for an infectious etiology based on Gram’s and PAS stainings. Immunohistochemical labeling revealed a predominance of B cells in idiopathic, bilateral necrotizing scleritis. Tinctorial staining abnormalities with Masson’s trichrome stain were present in scleral collagen of the two cases with idiopathic necrotizing scleritis as well as a case of secondary traumatic scleritis. Conclusions Based on a limited number of cases, idiopathic canine necrotizing scleritis shares similar histopathological features with non‐necrotizing scleritis and episcleritis; however, necrotizing scleritis is B‐cell‐dominated and bilateral, and significant collagen alterations manifest with Masson’s trichrome stain.     

绒山羊肥大细胞类胰蛋白酶的免疫组化及图像分析研究

[目的]探讨绒山羊肥大细胞中是否存在类胰蛋白酶及不同固定液对其常规染色和免疫组化染色结果的影响。[方法]采用兔抗羊肥大细胞类胰蛋白酶多克隆抗体间接免疫过氧化物酶技术检测由Carnoy液和中性福尔马林液(NBF)固定的绒山羊空肠、瓣胃和肺等组织肥大细胞中是否存在类胰蛋白酶。同时采用Image-ProPlus图像分析软件和人工计数法对结果进行分析,比较经常规染色和免疫组化染色后不同固定液固定组织中肥大细胞的形态及数量,进而判断Carnoy液和中性福尔马林液(NBF)对该检测技术的影响。[结果]兔抗羊多克隆抗体与绒山羊肥大细胞中的类胰蛋白酶具有良好的交叉反应,证实绒山羊肥大细胞胞浆颗粒中存在类胰蛋白酶。同时,与Carnoy液相比较,NBF液固定组织能较好地反映肥大细胞在组织中的数量和形态变化。[结论]绒山羊肥大细胞中存在类胰蛋白酶,且与Carnoy液相比较,NBF液是一种更为适合绒山羊肥大细胞常规染色和免疫组化染色的固定剂。     

J亚群白血病病理学观察及抗原定位

通过病理组织学和免疫组织化学技 术对1日龄感染鸡定期进行病理组织学观察 及gp85病毒糖蛋白检测,确定病理变化和J 亚群白血病的组织学嗜性及二者的关系。结 果在3周龄时出现第1只发病鸡,以后所有 鸡发病且病变随着日龄的增加而加重,主要 在骨髓、肝脏、脾脏、肾脏、卵巢、心肌等组织 中出现瘤细胞增生,并有炎性反应;免疫组在 3周龄时,肝脏、心脏、肺脏、小肠、肾脏、卵巢、 骨骼肌开始出现阳性反应,抗原主要在胞浆 中出现,呈深蓝色颗粒。在10周龄～14周龄 时,骨髓呈中度的阳性反应,而其它组织的阳 性信号出现的程度未见大的变化;在23周龄 时,肝内肿瘤呈较强阳性反应,信号主要集中 在细胞核;整个实验过程中,法氏囊、脑始终 呈阴性反应。研究发现,骨髓及肿瘤的抗原 性较强,并且肿瘤的抗原性要强于骨髓;另外 还发现,在阳性反应强的组织中,病变较明 显,肿瘤细胞增生明显,即阳性反应的组织器 官易形成肿瘤转移灶。     

A Case of Metastatic Adrenocortical Carcinoma Diagnosed with Steroidogenic Factor-1 in a Sprague-Dawley Rat

This report describes the morphological and immunohistochemical characteristics of an adrenocortical carcinoma with distant metastasis in a Sprague-Dawley rat. Macroscopically, a single large mass was observed in the adrenal gland, and multiple nodules were noted in the lung, liver and thyroid. Histologically, the adrenal tumor consisted of a solid growth of eosinophilic round cells with nuclear atypia. Vascular invasion was present, and multiple metastatic lesions were also observed in the lungs, liver, and mediastinal lymph nodes. Immunohistochemically, the nuclei of these tumor cells were positive for Steroidogenic Factor-1 (SF-1). In the thyroid, tumor cells histologically resembling adrenal cells were immunohistochemically negative for SF-1 but positive for calcitonin; thus the lesion was diagnosed as thyroid C-cell carcinoma. From these results, the present case was diagnosed as adrenocortical carcinoma with distant metastases. SF-1 could be a valuable marker for the differential diagnosis of adrenocortical tumors versus other endocrine tumors such as C-cell carcinoma.     

N -Methyl- N -nitrosourea-induced Renal Tumors in Rats: Immunohistochemical Comparison to Human Wilms Tumors

N-Methyl-N-nitrosourea (MNU)-induced renal tumors in rats and Wilms tumors in humans were compared. Renal mesenchymal tumors (RMTs) and nephroblastomas (blastemal and epithelial components) in female Lewis rats treated with a single intraperitoneal injection of 50 mg/kg MNU at birth and Wilms tumors (blastemal, epithelial and mesenchymal components) in humans were analyzed for the expression of pancytokeratin (CK), vimentin, p63, α-smooth muscle actin (SMA), desmin, S-100, CD57, CD117/c-kit, Wilms tumor 1 protein (WT1) and β-catenin. The mesenchymal components of rat RMTs and human Wilms tumors expressed vimentin, SMA and β-catenin. The blastemal components of rat nephroblastomas and human Wilms tumors expressed vimentin, CD117/c-kit and β-catenin. The epithelial components of rat nephroblastomas and human Wilms tumors expressed vimentin and β-catenin. WT1 was expressed in different cellular components of rat tumors as compared with human Wilms tumors; the expression was seen in mesenchymal tumors and blastemal components of nephroblastomas in rats and epithelial components in human Wilms tumors. CK, p63 and CD57 were not expressed in rat RMTs or nephroblastomas, while CK and WT1 were expressed in epithelial components and CD57 was expressed in blastemal and epithelial components of human Wilms tumors. Rat and human tumors were universally negative for the expression of desmin and S-100. The immunohistochemical characteristics of rat renal tumors and human Wilms tumors may provide valuable information on the differences in renal oncogenesis and biology between the two species.     

Mycoplasma leachii causes bovine mastitis: Evidence from clinical symptoms,histopathology and immunohistochemistry

Twelve quarters of six lactating cows were inoculated with Mycoplasma leachii strain GN407 through intramammary ductal infusion; another 12 quarters were inoculated with heat-inactivated M. leachii culture medium as negative controls. Multidisciplinary procedures, including clinical assessment, etiology assessment, pathology and immunohistochemistry (IHC), were used to elucidate the pathogenicity of M. leachii in bovine mastitis. From post-inoculation days (PIDs) 3 to 9, 12 inoculated quarters developed mild to severe clinical mastitis and mammary tissue histopathological changes, including inflammatory cell infiltration and architectural destruction of mammary gland ducts. The M. leachii antigen was also detected by IHC in the mammary gland epithelial cells of the inoculated quarters as a weak signal on PID 6 and as a strong signal on PID 9. The control quarters also developed mild mastitis and histopathological changes on PID 9, and M. leachii was also detected by IHC. Throughout the experimental period, the quarters of the negative control cow were clinically and pathologically normal, and the M. leachii antigen was not detected. In conclusion, direct histological and immunohistochemical evidence confirmed that M. leachii causes clinical bovine mastitis through histopathological lesions induced by invasion of the pathogen into mammary gland cells and through inflammatory cell infiltration.