Immunohistochemical study of caveolin-1 and -2 in the rat retina

The expression of caveolin-1 and -2 in the retina was examined; Western blot analysis showed that both were present. Immunohistochemistry indicated that caveolin-1 was expressed in the majority of retinal layers, including the ganglion cell layer, inner plexiform layer, outer plexiform layer, and in the vascular endothelial cells of the retina. Caveolin-2 was primarily immunostained in the vessels, but in a few other elements as well. This is the first demonstration of caveolin differential expression in the retina of rats, and suggests that caveolin plays an important role in signal transduction in glial cells and neuronal cells.     

Uterine Carcinosarcoma in a 2-year-old Female Wistar Hannover GALAS Rat

Carcinosarcomas are rare tumors in humans as well as rats and most commonly occur in the uterus. Recently, we observed a case of incidental carcinosarcoma of the uterus in a female Wistar Hannover GALAS [BrlHan:WIST@ Jcl (GALAS)] rat at 2 years of age. Histopathologically, the tumor was characterized by an admixture of malignant epithelial and nonepithelial elements. The carcinomatous components represented a type of endometrial carcinoma, consisting of glandular and solid proliferation of large-sized tumor cells. Prominent mitoses and tumor cell invasion were observed. The sarcomatous components were characterized by multifocal proliferation of severe atypical cells with cartilage matrix and were diagnosed as chondrosarcoma. Transitions between carcinomatous and sarcomatous components were observed, and many tumor cells in the solid lesion showed immunohistochemical reactivity with both cytokeratin and vimentin. Based on these findings, this tumor was diagnosed as a uterine carcinosarcoma. This is the first report of uterine carcinosarcoma in Wistar Hannover GALAS [BrlHan:WIST@Jcl (GALAS)] rats.     

26例犬毛囊肿瘤的组织病理学诊断和免疫组织化学分析

本研究旨在为犬毛囊肿瘤的诊断提供最优的免疫标记物,提高肿瘤诊断的准确性,缩短肿瘤的病理学诊断时间,为犬毛囊肿瘤病的临床精准治疗提供帮助.作者收集了26例临床犬毛囊肿瘤病例,分别用免疫标记物CK5/6、CK8/18、CK19、P63、CD34、CD10、Vimentin对肿瘤样本进行免疫组织化学标记(IHC).26例犬毛...     

Spontaneous Extraskeletal Osteosarcoma in a Rabbit (Oryctolagus cuniculus): Histopathological and Immunohistochemical Findings

A spontaneously occurring subcutaneous mass in the left forelimb of a nine-year-old rabbit (Oryctolagus cuniculus) was examined histopathologically and immunohistochemically. Clinically, edema and hemorrhage were seen around the mass. No connection of the tumor mass to the appendicular skeleton was found. The tumor was arranged in a solid growth pattern and irregular bundles, and neoplastic cells were polygonal to spindle-shape. Osteoid (positive for osteocalcin) and multinucleated giant cells were diffusely or focally seen. Neoplastic cells were positive for vimentin, osterix and Ki-67, indicating the nature of osteoblasts with proliferating activity, but negative for α-smooth muscle actin, desmin or CD204. Based on these findings, a diagnosis of extraskeletal osteosarcoma was made, a very rare tumor both in laboratory and pet rabbits.     

Ghrelin在成年皖西白鹅小肠内的免疫组化定位研究

采用免疫组化SABC法并结合DAB显色技术研究了皖西白鹅小肠内Ghrelin阳性细胞及其分布。结果：①Ghrelin免疫阳性细胞主要位于小肠的黏膜层，而黏膜下层和肌层未见Ghrelin阳性细胞。这些阳性细胞，胞浆着色，主要有两类，一类为“闭合型”细胞，呈圆形或椭圆形，其顶部不露于腔面，另一类为“开放型”细胞，呈锥体型，细胞顶部达到腔面。②在十二指肠，“开放型”和“闭合型”两种阳性细胞均可观察到，而在空肠和回肠，主要为“闭合型”阳性细胞。产生Ghrelin的细胞在成年皖西白鹅小肠黏膜层有广泛的分布，揭示Ghrelin可能调节小肠的功能。     

清洁级成年SD大鼠肝脏自发病变的组织病理学研究

为了研究清洁级成年SD大鼠肝脏自发病变的组织病理学改变,选取3 000只清洁级成年SD大鼠,雌雄各半,取肝脏做组织病理学检查.结果表明,清洁级成年SD大鼠肝脏的自发病理变化有细胞水肿、脂肪变性、炎症细胞浸润、肝窦扩张、灶性坏死、肝内胆管增生、囊肿、肝细胞癌等,自发病理变化无性别差异(P＞0.05).研究结果丰富了清洁级成年SD大鼠肝脏自发病变的组织病理学资料.     

Ultrastructural and Immunohistochemical Studies on Uptake and Distribution of FITC-Conjugated PLGA Nanoparticles Administered Intratracheally in Rats

Polylactide-glycolide (PLGA) nanoparticles have been developed as pulmonary drug delivery carriers. To investigate their behavior, small- (d50 = 74 nm) and large-sized (d50 = 250 nm) FITC-conjugated PLGA nanoparticles were intratracheally administered to rats and were traced for 5, 30 and 60 minutes and 24 hours after administration (HAT). Immunohistochemically, a, FITC-positive reaction was observed in type-I alveolar epithelial cells (type-I AEC), endothelial cells and alveolar macrophages in the lungs from 5 minutes after treatment (MAT) to 24 HAT in both nanoparticle groups. In the kidneys, a positive reaction was observed in proximal tubular epithelial cells at 30 MAT; the reaction peaked at 60 MAT and was reduced at 24 HAT, while no positive reaction was seen in other sites. Ultrascructurally, the number of membrane-bound vesicles, which were approximately 70 nm in size and hard to distinguish from pinocytic vesicles, apparently increased in type-I AEC and endothelial cells at 5 MAT in the small-sized group, in comparison with the control group receiving physiological saline. The number of vesicles in the large-sized group was almost same as that in the control group. On the other hand, in both nanoparticle groups, lysosomes filled with nanoparticles appeared in alveolar macrophages from 30 MAT to 24 HAT. These results indicate that PLGA nanoparticles might be quickly transferred from the alveolar space to the blood vessel via type-I alveolar epithelial cells and excreted into urine, and that there is a threshold for particle size, less than approximately 70 nm in diameter, with regard to absorption through the alveolar wall.     

小鼠和山羊子宫内膜淋巴细胞的制备

应用不同的消化方案,比较分析了胰蛋白酶和胶原酶对小子宫及山羊子宫内膜组织的消化效果,结果证明,对小鼠子宫消化的最佳方案是：１２０ＩＵ．ｍＬ＾－１Ｉ型胶原酶３７℃消化４ｈ,继之４℃消化１０ｈ,收获已消化细胞后,对残余组织利用２．５ｇ．Ｌ＾－１胰蛋白酶热消化５－１０ｍｉｎ,之后迅速加入５％左右的ＦＣＳ;对山羊子宫内膜的最适消化条件是１２０ＩＵ．ｍＬ＾－１Ｉ型胶原酶３７℃消化４ｈ,继之４℃消化１０ｈ;应用上述方法消化后,所获淋巴细胞活率≥９５％。     

Immunohistochemical Characterization of a Renal Nephroblastoma in a Trp53-mutant and Prolyl Isomerase 1-deficient Mouse

A nephroblastoma is a tumor arising from metanephric blastema occurring in childhood. Among laboratory rodents, nephroblastoma has been frequently reported in rats, but it remains exceedingly rare in mice. The present work describes a nephroblastoma in a young mouse homozygous for the specific Trp53 R172H point mutation coupled with targeted deletion of the Pin1 gene. The affected kidney was effaced by a biphasic tumor with an epithelial component arranged in tubules surrounded by nests of blastemal cells. Immunohistochemically, the neoplasm was diffusely positive for Wilms’ tumor antigen. The epithelial component expressed markers of renal tubular differentiation including wide-spectrum cytokeratin, E-cadherin and folate-binding protein. Furthermore, the neoplasm exhibited a high proliferative index and diffuse nucleocytoplasmic β-catenin expression. Based on histological and immunohistochemical features, a diagnosis of nephroblastoma potentially associated with Trp53 loss and oncogenic β-catenin activation has been proposed.     

Immunohistochemical evaluation of hepatic progenitor cells in different types of feline liver diseases

Hepatic progenitor cells are periportally resident cells capable of differentiating into mature hepatocytes or cholangiocytes to ensure hepatic regeneration. This reaction is termed a ductular reaction. In the present study, regenerative response of the feline liver to different hepatic diseases was investigated immunohistochemically. Regeneration of the liver through hepatocellular replication and proliferation of progenitor cell compartment were comparatively evaluated. Histological and immunohistochemical stainings were conducted on feline liver samples (n=40) representing various hepatobiliary diseases. Cytokeratin (CK) 7, CK19, Proliferating cell nuclear antigen (PCNA), Ki67, and Human hepatocyte marker 1 (Hep Par-1) were used. The presence of progenitor cells within feline livers was proved, both as passive cells in normal liver and as active cells (ductular reaction) in hepatic lesions. CK7 was found to be a suitable antibody for immunohistochemically detecting feline progenitor cells. In acute events, regeneration was predominantly shaped by the division of hepatocytes. In chronic events and severe acute events, hepatocytes lost their ability to divide and regeneration mainly occurred through progenitor cells. Location of the ductular reaction varied between different hepatic diseases. Parenchymal ductular reaction was detected in fulminant hepatitis, chronic hepatitis, hepatocellular lipidosis and metastatic lymphoma, whereas septal ductular reaction was detected in chronic hepatitis and metastatic lymphoma. Ductular reaction exhibited positive staining for Hep Par-1 in chronic and severe acute events. This study indicates the major role played by hepatic progenitor cells in regeneration of the feline liver. Moreover, it shows how the activation pattern of ductular reaction varies according to the hepatobiliary disease type.     

Age Difference in Morphology and Immunohistology inthe Thymus and Spleen in Crl:CD (SD) Rats

We investigated chronological changes in immunohistochemical phenotyping in the thymus and spleen in Crl:CD rats up to the age of about one year. In the thymus, T cells increased markedly from 3 to 4 weeks of age. Proliferating cells also increased markedly at these points. B cells tended towards an increase with age. In the spleen, white pulp increased until 9 weeks of age and remained fairly stable thereafter. In the periarteriolar lymphoid sheath and marginal zone, T cells gradually increased until 9 weeks of age and became almost flat thereafter. In the lymph follicle, T cells increased with age. B cells tended towards an increase with age in all areas of the spleen. It was concluded that development of the thymus was most marked from 3 to 4 weeks of age and that both the thymus and spleen had matured by 9 weeks of age.     

Immunohistochemical and histoplanimetrical study on the endothelial receptor involved in transportation of minute chylomicrons into subepithelial portal blood in intestinal villi of the rat jejunum

A portion of the minute chylomicrons less than 75 nm in diameter are transcytosed from the extravascular tissue into the subepithelial blood capillaries (sBC) in the villous apices of the rat jejunum. However, the details of the transportation mechanism have not been clarified. In this study, the endothelial receptor involved in the transportation of minute chylomicrons into the sBC’s lumina was immunohistochemically and histoplanimetrically examined in intestinal villi of the rat jejunum. Immunopositivity for very low density lipoprotein (VLDL) receptor was detected on the luminal and basal surfaces of the endothelial cells of sBC in approximately 68% of those apices of jejunal villi that possessed numerous chylomicrons in the lamina propria, while VLDL receptor was detected on the endothelial cells of sBC in only approximately 8% of intestinal villi that possessed few or no chylomicrons in the lamina propria. No immunopositivity for LDL receptor was detected in the sBC of all intestinal villi. These findings suggest that VLDL receptor is expressed by the endothelial cells of the sBC in conjunction with the filling of the lamina propria of jejunal villi with many chylomicrons produced by the villous columnar epithelial cells and that the VLDL receptor mediates the transportation of minute chylomicrons, maybe VLDL, into the subepithelial portal blood from the extravascular tissue of the rat jejunal villi.     

Spontaneous Adenosquamous Carcinoma with Rapid Growth and EMT-like Changes in the Mammary Gland of a Young Adult Female BALB/c Mouse

This study histopathologically and immunohistochemically investigated a spontaneously occurring single mass subcutaneously located in the left lower abdomen of a female BALB/cAJcl−nu/+ mouse at 10 weeks of age. The mass was about 20 × 15 × 10 mm in size after formalin fixation; nevertheless, it was not detected by clinical observations at 9 weeks of age. H&E staining revealed the tumor origin was epithelial and probably arose from the mammary gland, and the tumor cells demonstrated a squamous, acinar or polyhedral/basal pattern. A cell kinetics analysis revealed that many of the tumor cells of the squamous, acinar or polyhedral/basal component were positive for PCNA and cyclin D1, although there were a few of TUNEL-positive tumor cells in all of the components. An epithelial/mesenchymal analysis demonstrated that most of the tumor cells of the squamous and acinar components contained keratin and E-cadherin; however, most of the tumor cells of the polyhedral/basal component were less or very weakly positive for these markers. The tumor cells of the squamous component were negative for vimentin and SMA; however, many of the tumor cells of the polyhedral/basal component exhibited vimentin. In addition, expression of SMA was confirmed in some tumor cells of the acinar and basal components. Based on the microscopic and immunohistochemical characterizations, the tumor was diagnosed to be adenosquamous carcinoma that originated from the mammary gland with rapid growth, and the tumor cells demonstrated epithelial-mesenchymal transition-like changes.     

Availability of a Microglia and Macrophage Marker,Iba-1, for Differential Diagnosis of Spontaneous Malignant Reticuloses from Astrocytomas in Rats

In rats, it is sometimes difficult to distinguish malignant reticuloses from astrocytomas in routine histopathological assessment. In the present study, four spontaneous brain neoplasms developing in the cerebrum of one Wistar Hannover rat and three Sprague-Dawley rats were immunohistochemically examined using microglia and macrophage markers. Histopathologically, these neoplasms were localized mainly in the cerebral cortex, hypothalamus or piriform lobe, and the portions showing solid growth did not show characteristic cellular arrangement but had an indistinct boundary with the surrounding brain parenchyma. Neoplastic cells had oval or pleomorphic small nuclei with abundant eosinophilic cytoplasm. Two cases showed neoplastic cell infiltration into the meninges and perivascular spaces. Silver staining showed lack of reticulin fiber production in the stroma of the neoplasms. Immunohistochemically, the neoplastic cells were strongly positive for Iba-1 and sporadically positive for CD68 in all four cases. On the basis of these results, all the neoplasms examined here could be distinguished from astrocytomas and diagnosed as malignant reticuloses. Thus, immunohistochemical demonstration of microglia/macrophage characters, such as using Iba-1, is considered to be helpful for differential diagnosis of malignant reticuloses from astrocytomas among spontaneously occurring primary brain neoplasms in rats.     

Studies on Some Aspects of Neutrophil Functions and Uterine Defenses in Cows during the Estrous Cycle

Some aspects of the bovine uterine defense mechanisms were evaluated over two successive estrous cycles in five adult nonpregnant Friesian cows. Uterine flushings and blood samples were taken from each cow during the estrus and diestrus phases of each estrous cycle. A significant higher percentage of Candida albicans (C. albicans) were phagocytosed (P < 0.01) and killed (P < 0.05) by blood neutrophils during estrus than during diestrus. N-acetyl-β-D-glucosaminidase activity of uterine flushings, blood neutrophil alkaline phosphatase activity scores, and serum progesterone concentrations during diestrus increased significantly (P < 0.01) compared to those at estrus. Mean total blood leucocyte and monocyte counts increased significantly (P < 0.05) during estrus compared to diestrus .
It is concluded, that the different hormonal status has at each stage of the estrous cycle a definite effect on uterine resistance against bacterial infection and a fall in serum progesterone concentrations is associated with an increased ability of blood neutrophils to phagocytose and kill C. albicans .     

电刺激发情期大鼠蓝斑核对子宫平滑肌电活动的影响

在急性实验条件下,以爆发波(BW)的频率、波宽、峰电位频率为指标,观察了电刺激蓝斑核对发情期大鼠子宫平滑肌电活动的影响.结果:电刺激蓝斑核后,子宫角平滑肌BW频率增加极显著,波宽及峰电位频率虽有增加,但无显著差异;子宫颈平滑肌BW频率无显著增加,波宽却显著缩短,峰电位频率有所减少,未见明显差异.结果表明,蓝斑核对发情期大鼠子宫平滑肌电活动具有调节作用.     

无菌大鼠深二度烫伤感染模型机体炎性反应的研究

为探究无菌大鼠深二度烫伤感染模型机体炎性反应,试验利用无菌大鼠分别建立深二度烫伤损伤模型和烫伤后绿脓杆菌感染模型,比较这两种模型和SPF级大鼠深二度烫伤模型在愈合过程、炎性反应及病理变化等方面的异同。20只6周龄雌性Wistar无菌大鼠,在超净工作台内采用恒温恒压烫伤仪94℃作用8 s造成大鼠深二度烫伤,随机分为2组,一组伤口感染浓度为1×10⁵ CFU/mL绿脓杆菌菌液0.5 mL,一组不做处理,创面保持无菌状态,SPF级Wistar大鼠在同样条件下烫伤,创面不做处理。分别观察3组大鼠的伤口结痂时间、脱痂时间和愈合时间,并在烫伤前（0 h）,烫伤后24 h、3 d和7 d时,取血清,检测炎性因子肿瘤坏死因子-α（TNF-α）、白细胞介素1α（IL-1α）、粒细胞-巨噬细胞刺激因子（GM-CSF）的变化。结果显示,感染绿脓杆菌大鼠组开始结痂所需时间显著长于SPF级大鼠组和无菌大鼠组（P<0.05）;而脱痂及愈合所需时间显著变短（P<0.05）;感染绿脓杆菌组大鼠烫伤24 h时,皮下组织中可见淋巴细胞,有慢性炎性反应;3 d时,真皮深层有化脓性感染,横纹肌周围可见大量炎细胞,烫伤7 d时,皮下组织中有轻微血管扩张充血,化脓性感染减轻,无菌烫伤大鼠与SPF级烫伤大鼠皮肤在烫伤后一周内有轻度炎性反应。3组大鼠烫伤后,血清GM-CSF、IL-1α及TNF-α水平均较烫伤前显著升高（P<0.05）;烫伤24 h时,感染绿脓杆菌大鼠组和无菌大鼠组血清IL-1α水平显著高于SPF级大鼠组（P<0.05）;无菌大鼠组血清GM-CSF水平显著高于SPF级大鼠组和感染绿脓杆菌大鼠组（P<0.05）;烫伤3 d时,染绿脓杆菌大鼠组和无菌大鼠组血清IL-1α水平显著高于SPF大鼠组（P<0.05）,无菌大鼠组血清GM-CSF水平显著高于染绿脓杆菌大鼠组（P<0.05）。综上所述,无菌大鼠对烫伤刺激反应迅速强烈,血清中炎性因子迅速升高,SPF级大鼠深二度烫伤后炎性反应缓慢;无菌大鼠深二度烫伤伤口感染1×10⁵ CFU/mL绿脓杆菌,脱痂时间和愈合时间反而缩短。