THE RADIOGRAPHIC APPEARANCE OF PRIMARY LIVER NEOPLASIA IN DOGS

The signalment, anamnesis, and histopathologic, gross pathologic, and radiographic findings in 22 dogs with nonvascular, nonhematopoletic primary liver tumors were reviewed. The tumor types represented were hepatoma (8), bile duct cystadenoma (1), hepatocellular carcinoma (5), and cholangiocellular carcinoma (8). The dogs averaged 11.1 years of age. Females were predisposed to cholangiocellular carcinoma. The most common presenting clinical signs were general malaise, anorexia, PU/PD, vomiting, and seizures. Tumors ranged in size from diffuse 0.5-1 cm nodules to an 18-cm solitary mass and were located in any of the liver lobes. Four of the five diffuse tumors were cholangiocellular carcinomas. The most common radiographic appearance for any type of liver tumor was a right cranial abdominal mass causing caudal and left gastric displacement. In 54.5% of the dogs, radiographic evidence of intraperitoneal disease was identified. Nodular interstitial pulmonary masses were seen in 3 of the 22 dogs.     

Immunohistochemical characterization of canine hyperplastic hepatic lesions and hepatocellular and biliary neoplasms with monoclonal antibody hepatocyte paraffin 1 and a monoclonal antibody to cytokeratin 7.

Immunostaining with monoclonal antibody (MoAb) hepatocyte paraffin 1 (Hep Par 1) and an MoAb to cytokeratin 7 (CK7) was performed on 105 formalin-fixed, paraffin-embedded canine hyperplastic and neoplastic hepatic lesions. Hep Par 1 was detected in 12/12 hyperplastic nodules, 17/17 hepatocellular adenomas, and 37/40 hepatocellular carcinomas. The staining was disseminated, granular, and cytoplasmic. This antibody did not react with normal or neoplastic biliary epithelium. Other hepatic tumors or tumors metastatic to the liver did not bind Hep Par 1 except one metastatic intestinal carcinoma. MoAb to CK 7 stained all hyperplastic biliary epithelium and benign cholangiocellular tumors (5/5) and 14/18 cholangiocellular carcinomas. One hepatocellular carcinoma had cells positive for both Hep Par 1 and CK 7. Liver was the only normal tissue tested that reacted with MoAb Hep Par 1. Only five nonhepatic tumors (one adrenocortical carcinoma, one interstitial cell tumor of the testis, one melanoma, and two salivary adenocarcinomas) of 277 tumors tested had focal/multifocal staining for Hep Par 1. Prolonged fixation did not alter the staining with Hep Par 1. We conclude that Hep Par 1 is a specific and sensitive marker for canine hepatocellular tumors and allows distinction between hepatocellular and biliary neoplasms.     

Hepatocholangiocellular carcinoma in a rat--case report

A mixed epithelial tumour in the liver of a 24-month-old male Wistar rat from a 30-month inhalation study is described. The rat, which was in a group exposed to low concentrations of diesel exhaust, was euthanized because of emaciation, forced respiration and abnormal gait. Macroscopic examination of the enlarged liver revealed multiple partly confluent beige-red nodules up to 1.5 cm in diameter. Small nodules up to 7 mm in diameter were seen in the spleen. Histologically, the tumour nodules in the liver consisted of hepatocellular and cholangiocellular components. The hepatocellular component consisted of moderately differentiated polygonal to round hepatocytes about twice as large as normal hepatocytes and having hyperchromatic, centrally located nuclei with prominent nucleoli and eosinophilic cytoplasm. Foci of haematopoiesis and focal necroses were prominent. The cholangiocellular component was moderately differentiated and consisted of tubular structures lined by low cuboidal to cylindrical cells showing cytoplasmic basophilia and small dark nuclei without prominent nucleoli. The histological features of the nodules in the spleen corresponded to those of the primary tumour in the liver. Based on these criteria, the tumour nodules were diagnosed as hepatocholangiocellular carcinoma. The immunohistological examination confirmed the diagnosis, i.e. immunostaining for cytokeratins was positive for eight and 18 (hepatocellular carcinoma) and for seven and 19 (cholangiocellular carcinoma) as well as for vimentin (dense fibrous stroma). This tumour is considered to be spontaneous because of its single occurrence.     

Acquired myasthenia gravis and cholangiocellular carcinoma in a dog

Acquired myasthenia gravis and cholangiocellular carcinoma were diagnosed in a 7-year-old English Setter referred because of forelimb lameness, exercise-induced weakness, and fever. Three months earlier, the dog had had a pleuropulmonary infection caused by a Fusobacterium sp. The concurrent development of myasthenia gravis and cholangiocellular carcinoma in this dog may be explained by a paraneoplastic syndrome, although it is unproven. The cholangiocellular carcinoma may have possessed an acetylcholine receptor-like antigen on the tumor surface, which induced autoantibodies to cross-react with acetylcholine receptors at the neuromuscular junction.     

Isolation and characterization of canine tumor endothelial cells

The present study involved the isolation and characterization of canine tumor endothelial cells (TECs) from 2 malignancies. TECs were isolated using magnetic cell sorting following FITC labeling with UEA1 lectin, and they were characterized by measuring genetic and histopathological endothelial markers. Isolated TECs exhibited a cobblestone-like morphology and expressed both vascular endothelial growth factor receptor 2 (VEGFR2) and Von Willebrand factor (vWF). Further, both TECs and tumor cells derived from a seminoma exhibited increased C-X-C chemokine receptor type 7 (CXCR7) expression. However, CXCR7 expression was not detected in TECs and tumor cells derived from a hepatocellular carcinoma. Understanding TEC specific traits may be important in the development of more efficacious anti-angiogenic therapies that do not induce adverse effects.     

Identification of hepatic stem/progenitor cells in canine hepatocellular and cholangiocellular carcinoma

Hepatic progenitor cells (HPCs) are bipotential stem cells residing in human and animal livers that are able to differentiate towards the hepatocytic or cholangiocytic lineages. HPCs are present in both hepatocellular (HCC) and cholangiocellular carcinoma (CC) in humans; and a small percentage of HCC can originate from cancer stem cells. However, its distribution in canine liver tumour has not been studied. Herein, we searched for stem/progenitor cells in 13 HCC and 7 CC archived samples by immunohistochemical analysis. We found that both liver tumours presented a higher amount of K19‐positive HPCs. Besides, 61.6% of HCC cases presented immature CD44‐positive hepatocytes. Nevertheless, only two cases presented CD133‐positive cells. As observed in humans, hepatic canine tumours presented activated HPCs, with important differentiation onto hepatocytes‐like cells and minimal role of cancer stem cells on HCC. These findings reiterate the applicability of canine model in the search for new therapies before application in humans.     

Serum alpha-fetoprotein values in dogs with various hepatic diseases.

Serum alpha-fetoprotein (AFP) values were measured in hepatic diseased dogs with or without tumor and non-hepatic tumor bearing dogs by a sandwich ELISA using anti-dog AFP antiserum. Serum AFP values were less than 70 ng/ml in clinically healthy dogs. The values in dogs with hepatocellular carcinoma were higher than 1,400 ng/ml in 7 of 9 dogs, wherever those in two dogs with cholangiocarcinoma were in the normal range. Serum AFP values in hepatic diseased dogs without tumor were also high, however, the values were below 500 ng/ml in 90% of the dogs. In non-hepatic tumor dogs, serum AFP values were less than 500 ng/ml in 76% of the dogs. In the surgically removal cases with hepatocellular carcinoma, serum AFP values rapidly decreased. These results suggested that the sandwich ELISA using anti-dog AFP antiserum was an available method for diagnosis of hepatocellular carcinoma in dogs.     

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.     

Hepatocellular carcinoma associated with erythrocytosis and hypoglycemia in a yearling filly

A yearling Arabian-type filly with a history of poor growth, erythrocytosis, hypoglycemia, and high liver enzyme activities was admitted to the hospital for evaluation. Three days after admission, the filly collapsed, deteriorated rapidly despite treatment, and was euthanatized. A metastatic hepatocellular carcinoma with capsular rupture and hemoperitoneum were found at necropsy. Primary liver tumors are rare in horses, and hepatocellular carcinoma has been reported in only 1 other horse. The systemic manifestations of the tumor in this filly included weakness, weight loss, inappetence, erythrocytosis with tumor production of erythropoietin, persistent hypoglycemia with normal serum insulin concentrations, serum alpha-fetoprotein (normally present only during fetal life), and terminal massive hemoperitoneum, all features of the syndrome in man.     

Antemortem diagnosis of hepatic adenocarcinoma in a ewe

A 4-year-old ewe with weight loss, anorexia, and high liver enzyme activities was diagnosed at necropsy as having cholangiocellular carcinoma that had metastasized to kidneys, adrenal glands, heart, lungs, mediastinum, and peritoneal surface of the diaphragm. Cholangiocellular carcinoma is an uncommon neoplasm of sheep. Evaluation of liver tissue obtained via ultrasound-guided percutaneous liver biopsy had suggested an antemortem diagnosis of adenocarcinoma of the liver.     

Four cases of neoplasia in captive wild birds

Four cases of malignant neoplasia in captive wild birds are described: an adenocarcinoma of the adrenal gland in a Mountain duck (Tadorna tadornoides), a malignant melanoma in the thoracic cavity of a Combed duck (Sarkidiornis melanotos), a hepatocellular carcinoma with pulmonary metastasis in an Asian Purple Swamphen (Porphyrio porphyrio), and an undifferentiated carcinoma in the abdomen with metastasis to skeletal muscle in a White-Breasted Waterhen (Amaurornis phoenicurus). The tumors were diagnosed during a 1-year period and represented an incidence of neoplasia of 3.1%. These appear to be the first documented cases of a malignant adrenal gland tumor and a non-ocular melanoma in the order Anseriformes. The hepatocellular carcinoma failed to react with an immunoperoxidase stain for alphafetoprotein.     

Alpha-fetoprotein in serum and tumor tissues in dogs with hepatocellular carcinoma.

Serum alpha-fetoprotein (AFP) concentrations were measured before and after surgical removal of tumor masses in four dogs with hepatocellular carcinoma (HCC). Localization of AFP was also examined immunohistochemically in tumor tissues. In three cases, the serum AFP concentration was 10 to 20 times higher than that of normal dogs. One to two months after surgery, the serum AFP concentration had decreased to normal range. AFP was localized in the tumor tissues in these three cases. One case, which had a low serum AFP, did not show AFP localization in tumor tissue.     

A multistep approach in the cytologic evaluation of liver biopsy samples of dogs with hepatic diseases

Cytologic criteria were evaluated for their diagnostic value in liver disease in dogs. Therefore, histopathologic and cytologic examination was performed on liver biopsy samples of 73 dogs with liver diseases and 28 healthy dogs. Logistic regression analysis was used to select the measured parameters to be included in a multistep approach. With the logistic regression method, different characteristic cytologic parameters could be defined for each histopathologic diagnosis. In malignant lymphoma of the liver, the presence of large numbers of lymphoblasts with a minimum of 5% of all cells was found. Clusters of epithelial cells with several cytologic characteristics of malignancy intermixed with normal hepatocytes were indicative of metastatic carcinoma or cholangiocellular carcinoma. Liver cells in hepatocellular carcinoma were characterized by a high nucleus/cytoplasm ratio, large cell diameters, increased numbers of nucleoli per nuclei, small numbers of cytoplasmic vacuoles, and frequently, small numbers of lymphocytes. Extrahepatic cholestasis was characterized by excessive extracellular bile pigment in the form of biliary casts, an increased number of nucleoli within hepatocytes, decreased hepatic cell size, and low numbers of lymphocytes. In destructive cholangiolitis, increased numbers of neutrophils and a small mean nuclear size within hepatocytes were seen. Acute and nonspecific reactive hepatitis are diagnosed based on the presence of moderate reactive nuclear patterns, including more pronounced chromatin, prominent nucleoli, increased numbers of inflammatory cells, excluding lymphocytes, and the absence of increased numbers of bile duct cell clusters. Increased number of mast cells also was indicative of nonspecific reactive hepatitis. Important cytologic criteria for the diagnosis of liver cirrhosis, in addition to chronic hepatitis, are intracellular bile accumulation and increased numbers of bile duct cell clusters. In summary, the stepwise approach based on logistic regression presented in this study might be helpful in the objective cytologic diagnosis of hepatic diseases.     

肝癌细胞靶向肽的筛选与鉴定

利用噬菌体环七肽库筛选与肝癌细胞特异性结合的多肽,并对其亲和力进行生物学鉴定。以HL-7702为消减细胞,HepG2为筛选靶细胞,对噬菌体随机环七肽库进行4轮全细胞消减筛选,并随机挑取60个阳性噬菌体克隆,以ELISA法鉴定其与HepG2细胞的结合活性,并取阳性克隆进行测序分析,并合成多肽进行免疫细胞化学染色鉴定。经4轮筛选,噬菌体在靶细胞HepG2上出现明显富集;利用ELISA从随机挑选的60个噬菌体克隆中得到15个与肝癌细胞具有高结合力的阳性克隆,测序并进行序列分析比对发现氨基酸序列无同源性,经免疫细胞化学染色鉴定后,发现1条多肽序列亲和力较高,为提高抗菌肽对肿瘤细胞的靶向杀伤作用奠定基础。     

Chemotherapy Versus Chemotherapy Plus Radiotherapy in the Treatment of Tonsillar Squamous Cell Carcinoma in the Dog

The effect of adjuvant chemotherapy and combined chemotherapy and radiotherapy in 22 dogs with advanced canine tonsillar carcinoma (World Health Organization [WHO], T2-3 primary tumor with infiltration into surrounding tissues) was evaluated. There were four treatment groups of at least five dogs each. Combinations of chemotherapeutic drugs reported to have activity against squamous cell carcinoma in humans and dogs (doxorubicin, cisplatin, vinblastine, and cyclophosphamide) were administered after tonsillectomy. Radiation therapy (orthovoltage type, external beam) was combined with chemotherapy in one treatment group. The dogs that had combination radiation therapy and chemotherapy had higher response rates and significantly longer survival times. However, most dogs died of progression of disease.     

Arginase-1 and P-glycoprotein are downregulated in canine hepatocellular carcinoma

BackgroundHepatocellular carcinoma is the most common primary hepatic malignancy in humans and dogs. Several differentially expressed molecules have been studied and reported in human hepatocellular carcinoma and non-neoplastic liver lesions. However, studies on the features of canine hepatocellular carcinoma are limited, especially related to the differential characteristics of neoplastic and non-neoplastic lesions.ObjectivesThe study''s objective was 1) to examine and evaluate the expression of arginase-1, P-glycoprotein, and cytokeratin 19 in canine liver tissues and 2) to investigate the differential features of hepatocellular carcinomas, liver tissue with non-neoplastic lesions, and paracancerous liver tissues in dogs.MethodsThe expression levels of three markers underwent immunohistochemical analysis in 40 non-neoplastic liver tissues, 32 hepatocellular carcinoma tissues, and 11 paracancerous liver tissues. Scoring of each marker was performed semi-quantitatively.ResultsArginase-1 and P-glycoprotein were significantly downregulated in hepatocellular carcinoma, compared with hepatic tissues with non-neoplastic diseases (p < 0.001). Expression levels of arginase-1 and P-glycoprotein were also significantly lower in hepatocellular carcinoma than in paracancerous liver tissues (arginase-1, p = 0.0195; P-glycoprotein, p = 0.047). Few cytokeratin 19-positive hepatocytes were detected and only in one hepatocellular carcinoma and one cirrhotic liver sample.ConclusionsThe results of this study suggest that downregulation of arginase-1 and P-glycoprotein is a feature of canine hepatocellular carcinoma; thus, those markers are potential candidates for use in differentiating hepatocellular carcinomas from non-neoplastic liver lesions in dogs.     

Vimentin/cytokeratin coexpression foci in a well-differentiated canine hepatocellular carcinoma

A number of pale-stained cell foci were observed within a well-differentiated hepatocellular carcinoma which developed in a 10-year-old male mongrel dog. The foci were composed of hepatocyte-like cells, but did not contain glycogen granules in their cytoplasm. Immunohistochemically, the focus cells coexpressed both bile duct type cytokeratin and vimentin. Electronmicroscopically, they were abundant in cytoplasmic organelles and contained bile pigments. Bile canaliculi were noted between the focus cells. The focus cells in the present case were considered to be neoplastic hepatocytes expressed bipotential features of hepatic stem cells.     

Hepatobiliary neuroendocrine carcinoma in cats: a clinicopathologic, immunohistochemical, and ultrastructural study of 17 cases

Hepatobiliary neuroendocrine carcinoma was diagnosed in 17 cats in a period of 10 years. Seven tumors were of intrahepatic origin, one of which was a composite containing components of epithelial and neuroendocrine carcinoma. Nine tumors were of extrahepatic origin, and one tumor was located in the gall-bladder. The cats were adult and geriatric, and the male : female ratio varied according to tumor group. Hepatomegaly, anorexia, weight loss, and vomiting were the most common clinical signs observed in the cats with hepatic neuroendocrine carcinoma. The cats with extrahepatic neuroendocrine carcinoma showed these signs plus icterus (5/9) and high concentrations of hepatic enzymes. Histologically, the hepatic neuroendocrine carcinomas had two patterns, one with acinar structures separated by vascular stroma lined by cuboidal or columnar cells and the other solid with groups of anaplastic cells separated by vascular stroma. The composite tumor consisted of both bile duct carcinoma and neuroendocrine carcinoma. The extrahepatic neuroendocrine carcinomas and the gallbladder neuroendocrine carcinoma were characterized by solid sheets or groups of round to oval cells with vascular or fibrovascular stroma. Immunohistochemical examination of 10 of the neuroendocrine carcinomas revealed that all 10 stained with neuron-specific enolase; one bile duct carcinoma and the gallbladder carcinoma stained with chromogranin; four of five bile duct carcinomas and the gall bladder carcinoma stained with synaptophysin; and one bile duct carcinoma stained with gastrin. One cat with hepatic carcinoma had duodenal ulcer; in this cat, ultrastructural studies showed neurosecretory granules leading to the diagnosis of Zollinger-Ellison syndrome. In four cats in which necropsy was permitted, carcinomatosis (4/4), lymph nodes (4/4), lungs (2/4), and intestines (1/4) were the metastatic sites. Fourteen of the 17 cats were euthanatized during or immediately after surgery.     

Hypoglycemia associated with nonislet cell tumor in 13 dogs

Hypoglycemia associated with nonislet cell tumor was found in 13 dogs. In each dog, clinical signs were related directly to adrenergic and neuroglucopenic effects of hypoglycemia and included collapsing episodes, tremors, restlessness, weakness, and grand mal seizures that were responsive to glucose administration. Eight of the dogs had hepatocellular carcinoma; surgical resection of the tumor achieved remission of clinical signs in 3 of these dogs. Other hepatic tumors associated with hypoglycemia included leiomyosarcoma and hemangiosarcoma involving solitary lobes of the liver. Nonhepatic tumors included splenic hemangiosarcoma, diffuse metastatic melanoma, and salivary gland adenocarcinoma.