Aberrant expression of cyclin D1 in pulmonary proliferative lesions induced by high doses of urethane in transgenic mice carrying the human prototype c-H-ras gene

In our previous study, when rasH2 mice and non-transgenic (non-Tg) littermates were injected intraperitoneally with 1,000 mg/kg of urethane once or three times at two-day intervals, the incidence of lung proliferative lesions in rasH2 mice given triple doses of urethane was significantly increased, compared to that in rasH2 mice given a single dose of urethane, and the mutation frequency of the transgene in lung tumors in rasH2 mice given triple doses was lower than that in rasH2 mice given a single dose of urethane. In the present study, differential immunohistochemical expressions of Cyclin D1 and PCNA, that lead to abnormal cell proliferation and tumor development due to uncontrolled G1-S transition in the cell cycle, as well as p53 tumor suppressor gene in pulmonary proliferative lesions obtained from our previous study were investigated. Over-expression of Cyclin D1 in hyperplasias in rasH2 mice given triple doses was significantly increased, compared to that in the single-injection group, but no significant differences in Cyclin D1 between the single and triple injection groups were observed in hyperplasias in non-Tg mice or lung tumors in either rasH2 or non-Tg mice. There were no differences in the PCNA labeling index of hyperplasias in rasH2 or non-Tg mice between the triple-injection and single-injection groups, while the PCNA labeling index tended to be increased in the tumor, compared with that in hyperplasias. There was neither mutation of p53 nor an increase in immunoreactivity of wild type p53 in these proliferative lesions. These results suggest that cyclin D1 over-expression in alveolar/bronchiolar hyperplasias in rasH2 mice in the triple-injection group is not only indicative of a high cell proliferation rate but also of an important role in the process of malignant transformation.     

Cellular proliferative and telomerase activity in canine mammary gland tumors

In canine mammary tumors, we examined the telomerase activity, proliferative activity by proliferative cell nuclear antigen (PCNA) immunohistochemistry, and percentage of apoptotic cells by the deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method. The relationship between these measures and histopathologic malignancy was also investigated. PCNA index was highest in malignant tumors (adenocarcinoma: 27.0%; malignant mixed tumor: 15.7%), followed by benign tumors (adenoma: 4.4%; benign mixed tumor: 5.3%), hyperplasia (2.1%), and normal mammary gland (0.9%). In adenoma and adenocarcinoma, papillary and solid types showing higher cellularity tended to have higher PCNA indices than did cystic and tubular types. Although the TUNEL index was <1% in all cases, the relationship between this measure and histopathologic diagnosis showed the same tendency as observed in PCNA immunostaining. Telomerase activity was detectable in all adenomas, benign mixed tumors, and adenocarcinomas examined. In contrast, all normal mammary glands, hyperplasias, and malignant mixed tumors were negative for telomerase. Relative telomerase activity (RTA) of adenocarcinoma (56.5) was significantly higher than that of adenoma (27.8) and benign mixed tumor (33.9), and a significant positive correlation (P < 0.001) was noted between RTA and PCNA index. No significant correlations were noted between either PCNA or TUNEL index and clinical features such as metastasis and tumor diameter. PCNA index and telomerase activity may be useful markers for judging malignancy of canine mammary tumors.     

Primary epididymis squamous cell carcinoma in a CB6F1-Tg rasH2 mouse

In a 26-week carcinogenicity study in rasH2-Tg mice, squamous cell carcinoma on the epididymis was observed in a male mouse in the positive control group treated with N-methyl-N-nitrosourea. A 29-week-old male rasH2-Tg mouse that was euthanized 21 weeks after the administration of N-methyl-N-nitrosourea had a white-grayish mass on the left caput epididymis. The mass was nodular and consisted of pleomorphic tumor cells forming alveolar, sheeted, and trabecular structures suggesting epithelial tumor growth. These cells presented a cobblestone-like arrangement and formed intercellular bridges. Keratinization was infrequently observed. Periodic acid-methenamine-silver staining revealed argyrophilic fibrous structures around the alveolar structure of the tumor cells. Immunohistochemically, the tumor cells were positive for cytokeratin AE1/AE3 and cytokeratin 14 and negative for cytokeratin 5, p63, uroplakin III, vimentin, desmin, and αSMA. These immunohistochemical results suggested the tumor cells originated from the epididymal ducts. Metastatic lesions were observed in the mesenteric, inguinal, and pancreaticoduodenal lymph nodes. Based on these results, this tumor was diagnosed to be a primary squamous cell carcinoma of the epididymis. This is the first report of primary squamous cell carcinoma of the epididymis in a rasH2-Tg mouse.     

Survey of tumorigenic sensitivity in 6-month rasH2-Tg mice studies compared with 2-year rodent assays

The pharmacokinetic endpoint of a 25-fold increase in human exposure is one of the specified criteria for high-dose selection for 2-year carcinogenicity studies in rodents according to ICH S1C(R2). However, this criterion is not universally accepted for 6-month carcinogenicity tests in rasH2-Tg mice. To evaluate an appropriate multiple for rasH2-Tg mice, we evaluated data for 53 compounds across five categories of rasH2-Tg mouse-positive [(1) genotoxic and (2) non-genotoxic] carcinogens and rasH2-Tg mouse-negative [(3) non-genotoxic carcinogens with clear or uncertain human relevance; (4) non-genotoxic rodent-specific carcinogens; and (5) non-carcinogens], and surveyed their tumorigenic activities and high doses in rasH2-Tg mice and 2-year rodent models. Our survey indicated that area under the curve (AUC) margins (AMs) or body surface area-adjusted dose ratios (DRs) of tumorigenesis in rasH2-Tg mice to the maximum recommended human dose (MRHD) were 0.05- to 5.2-fold in 6 category (1) compounds with small differences between models and 0.2- to 47-fold in 7 category (2) including three 2-year rat study-negative compounds. Among all 53 compounds, including 40 compounds of the rasH2-Tg mouse-negative category (3), (4), and (5), no histopathologic risk factors for rodent neoplasia were induced only at doses above 50-fold AM or DR in rasH2-Tg mice except for two compounds, which induced hyperplasia and had no relationship with the tumors observed in the rasH2-Tg mouse or 2-year rodent studies. From the results of these surveys, we confirmed that exceeding a high dose level of 50-fold AM in rasH2-Tg mouse carcinogenicity studies does not appear to be of value.     

Cross-Sectional Imaging Characteristics of Pituitary Adenomas, Invasive Adenomas and Adenocarcinomas in Dogs: 33 Cases (1988–2006)

Background: Pituitary tumors in dogs can be adenomas, invasive adenomas, or adenocarcinomas. In people, invasive adenomas and pituitary adenocarcinomas carry a worse prognosis than adenomas.
Hypothesis/Objective: To identify differentiating features on cross-sectional imaging in dogs with pituitary adenomas, invasive adenomas, and adenocarcinomas.
Animals: Thirty-three dogs that had computed tomography (CT) or magnetic resonance imaging (MRI) performed and a necropsy diagnosis of pituitary adenoma ( n = 20), invasive adenoma ( n = 11), or adenocarcinoma ( n = 2).
Methods: Medical records were retrospectively reviewed for signalment, history, and diagnosis. CT and MR images were reviewed for characteristics of pituitary tumors.
Results: Mean (± standard deviation) age for dogs with pituitary adenomas (10.6 ± 2.9 years) was greater than that of those with invasive adenomas (8.3 ± 2.7 years, P = .04). Eighteen out of 20 (90%) dogs with adenomas had contrast-enhancing masses. Thirteen out of 20 (65%) had homogeneous enhancement. Mean adenoma height was 1.2 ± 0.7 cm. Eight out of 20 (40%) adenomas were round and 8/20 (40%) compressed surrounding brain. Eleven out of 11 dogs (100%) with invasive adenomas had contrast-enhancing masses. Seven out of 11 (64%) masses were homogeneous. Mean invasive adenoma height was 1.8 ± 0.7 cm, which was significantly greater than adenomas ( P = .03). Mass shape varied from round to oval to irregular. Six out of 11 (55%) masses compressed surrounding brain. Clinical and imaging features were variable for 2 dogs with adenocarcinomas.
Conclusions and Clinical Relevance: Invasive adenoma should be suspected if a dog with a pituitary tumor is <7.7 years of age and has a mass >1.9 cm in vertical height. Adenocarcinomas are uncommon and metastatic lesions were not seen with imaging.     

Characterization of spindle cell component of ferret (Mustela putorius furo) adrenal cortical neoplasms – correlation to clinical parameters and prognosis

Adrenal cortical epithelial tumours are common in ferrets. A variant tumour type with prominent spindle cell proliferation has been identified. We characterized these variant tumours with light microscopy and immunohistochemical analysis and correlate these features to clinical parameters and prognosis. We classified 24 ferret adrenal cortical masses with recognizable spindle cell proliferation obtained from the AMC and AFIP databases, based on percentage of spindle cells present and features of malignancy. These masses were separated into hyperplastic nodules and adenomas (both with 1–24% spindle cells), ‘mixed’ adenomas (≥25% spindle cells), adenocarcinomas (1–24%) and ‘mixed’ adenocarcinomas (≥25% spindle cells). Tumours were evaluated immunohistochemically for smooth muscle actin (SMA) and estrogen receptor (ER) expression. Disease‐free interval (DFI) and survival time (ST) were calculated using Kaplan–Meier product limit method. Of 24 cases of spindle cell variant adrenal tumours, one was a hyperplastic nodule, 10 were adenomas, three were ‘mixed’ adenomas, six were adenocarcinomas and four were ‘mixed’ adenocarcinomas. The proliferative spindle cell cytoplasm was SMA‐positive (smooth muscle myocyte origin). ER positivity, seen in nine of 24 cases, was restricted to adenocarcinomas, ‘mixed’ adenomas and ‘mixed’ adenocarcinomas. DFI and ST were significantly reduced in ‘mixed’ adenocarcinomas or tumours with ER expression. DFI was significantly reduced in tumours with marked smooth muscle. The spindle cell component of these variant adrenal tumours is smooth muscle in origin. The presence of abundant smooth muscle, a more malignant histologic grade (‘mixed’ adenocarcinomas) and ER expression are significantly positively correlated to both decreased DFI and decreased ST.     

Histopathological characterization of the skeletal myopathy in rasH2 mice carrying human prototype c-Ha-ras gene

A skeletal myopathy is found in approximately 100% of rasH2 mice. To confirm detailed features of the rasH2 skeletal myopathy, the biceps femoris, diaphragm, triceps brachii, gastrocnemial (types I and II fiber-mixed muscles) and soleus muscle (type I fiber-dominant muscle) obtained from male rasH2 and non-transgenic littermates aged 10-13 and 34 weeks were examined. Variations in the muscle fiber size, early-scattered degeneration/necrosis and regeneration of muscle fibers were detected in 10-13-week-old rasH2 mice. The severity of the above muscular lesions was more prominent in older rasH2 mice. These lesions were noted in the type II myofiber dominant muscles (biceps femoris, triceps brachii and gastrocnemial). NADH-TR stain clearly demonstrated a disorganized intermyofibrillar network and necrotic change in muscle fibers. No specific morphological changes, like rod structure or tubular aggregation seen in some types of myopathy, were noted in Gomori trichrome and NADH-TR stains in the rasH2 mouse like in many types of muscular dystrophy. Electronmicroscopically, occasional muscle fiber degeneration/regeneration, invaded phagocytic cells, indistinct Z-band suggesting excessive contraction and dilatation of the sarcoplasmic reticulum were observed. In summary, the skeletal myopathy occurring in rasH2 mice is consistent with muscular dystrophy characterized morphologically by progressive degeneration and regeneration of myofibers. The myopathy is confined to the type II myofiber predominant muscles and is not associated with any pathognomonic lesions. These characteristics will provide us with a useful model for research in muscular dystrophy of diverse myofibers.     

Cyclooxygenase-2 expression in canine mammary tumors

Mammary tumors are the most common neoplasms in female dogs. Induction of cyclooxygenase-2 (COX-2) has been implicated in various cancers in humans. However, expression of COX-2 has not been investigated in canine mammary tumors. Normal mammary gland (n = 4), simple or complex adenomas (n = 63), and simple or complex adenocarcinomas (n = 84) were studied by immunohistochemistry. Results showed that COX-2 was not expressed in the normal gland but was detected in 24% of adenomas and in 56% of adenocarcinomas (P < 0.001). The incidence of COX-2 expression and the intensity of the COX-2 signal were higher in adenocarcinomas than in adenomas (P < 0.001). These results demonstrate for the first time that COX-2 is induced in a proportion of canine mammary tumors and that COX-2 expression is more frequent and more intense in malignant than in benign tumors, suggesting a potential role for COX-2 in canine mammary tumorigenesis.     

rasH2 mouse: reproducibility and stability of carcinogenicity due to a standardized production and monitoring system

The rasH2 mouse was developed as a model for carcinogenicity studies in regulatory science. Its phenotype is stable during high-volume production and over successive generations. To produce rasH2 mice, three strains of mice (C57BL/6J-TgrasH2, C57BL/6J, and BALB/cByJ) were maintained individually. Since the homozygous c-HRAS genotype is lethal, hemizygous transgenic mice were maintained by crossing with inbred C57BL/6J mice. After breeding, male B6-transgenic mice were mated with female BALB/cByJ mice to obtain transgenic mice. Pups that were rasH2-Tg (tg/wt) or rasH2-Wt (wt/wt) were confirmed by genotyping. Frozen embryos were preserved by the Central Institute for Experimental Animals (CIEA) and sent to two facilities, CLEA Japan and Taconic Biosciences, where the mice were produced. Production colonies are created in both facilities and supplied to customers worldwide. To prevent genetic drift, the colonies were renewed for up to 10 generations, and renewals were carried out four times every five years from 2005 to 2021. To ensure the uniformity and maintenance of the phenotype of rasH2 mice, the carcinogen susceptibilities were monitored in every renewal of colonies by CIEA based on a standard protocol of the short-term carcinogenicity study using the positive control compound N-methyl-N-nitrosourea (MNU). Furthermore, simple carcinogenicity monitoring targeting the forestomach, the organ most sensitive to MNU, was performed approximately once a year. Based on the optimally designed production and monitoring systems, the quality of rasH2 mice with reproducibility and stability of carcinogenicity is maintained and supplied globally.     

Immunoadjuvant activity of crude lectin extracted from Momordica charantia seed

The aim of this study is to investigate the immunoadjuvant activity of the crude Momordica charantia lectin (crMCL) extracted from seed using beta-galactosidase (beta-gal) as the model antigen. BALB/c mice were injected intramuscularly with beta-gal alone or beta-gal + crMCL for up to four immunizations at two-week intervals. After administration of 2 doses, the IgG-specific titer to beta-gal was significantly higher in mice in the beta-gal + crMCL group than in that from the animals from the beta-gal alone group, while it was about the same in both groups after 1 dose. Our data suggest that crMCL may help raise antibodies under the prime and boost administration regimen and could be a potent vaccine adjuvant.     

Differential expression of cell cycle regulators p21, p27 and p53 in metastasizing canine mammary adenocarcinomas versus normal mammary glands

The cyclin dependent kinase inhibitors p21 and p27 are important regulators of cell cycle progression. To analyze their role in the malignant progression of canine mammary tumors expression levels of p27 and p21 and its major regulator p53 were compared in simple adenomas, adenocarcinomas of the mammary gland and lymph node metastases with normal mammary gland. Laser microdissection of tissue samples and real-time PCR were used for quantification of mRNA expression levels. p21 was overexpressed in adenocarcinomas, whereas adenomas and metastases expressed p21 more heterogeneously. Comparison of p21 expression in adenocarcinomas and their metastases revealed a significant decrease in expression in metastases. In contrast, p27 expression was reduced in the adenocarcinomas but heterogeneously expressed in adenomas and metastases. Taken together the results suggest that loss of p21 overexpression is associated with tumor metastasis while reduced cell cycle inhibition by p27 is associated with malignant progression.     

Immunohistochemical detection of Mdm2 and p53 in feline mammary gland tumors

The objective of this study was to evaluate nuclear reactivity of Mdm2 and p53 proteins by immunohistochemical means in feline mammary gland tumors; 12 adenomas which included 6 adenomatous lesions obtained from the tissue adjacent to adenocarcinomas, and 22 adenocarcinomas. Seven adenomas and 18 adenocarcinomas showed moderate or marked Mdm2 reactivity. Sixteen adenocarcinomas showed moderate to marked p53 reactivity, but 9 adenomas showed none. Discordant Mdm2 overexpression was found in 5 adenomas and 3 adenocarcinomas, although co-overexpression of Mdm2 and p53 was found in 15 adenocarcinomas. These results suggest that nuclear overexpression of Mdm2 is present in the tumors of early stage without p53 overexpression and related to feline mammary gland tumorigenesis. Nuclear overexpression of p53 is more frequent in adenocarcinomas, but not in adenomas.     

Quantitative analysis in spontaneous canine anal sac gland adenomas and carcinomas

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas. The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.     

Quantitative analysis in spontaneous canine anal sac gland adenomas and carcinomas

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas.The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.     

Chemical-induced lung tumor in Tg-rasH2 mice: a novel mouse tumor model to assess immune checkpoint inhibitors combined with a chemotherapy drug

In subcutaneous tumor models, changes in the tumor microenvironment can lead to differences in therapeutic treatment responses between the subcutaneous and parent tumors. Accordingly, we generated a lung carcinogenesis model that combines genetically modified mice (Tg-rasH2 mice) with two-stage chemical carcinogenesis as an alternative to the subcutaneous tumor model. In this model, Tg-rasH2 mice were treated with 1-ethyl-1-nitrosourea, followed by butylhydroxytoluene. Mice developed lung adenomas five weeks after treatment initiation. Subsequently, anti-mouse PD-1 antibody (α-mPD-1) or isotype control was administered intraperitoneally twice a week for 4 weeks. Tumor growth was examined by measuring the relative tumor area in serially sliced lung histopathological specimens. No statistically significant differences were observed in the relative lung tumor areas between treated and control groups. A second experiment then examined the antitumor efficacy of α-mPD-1 combined with gemcitabine in a mouse model. Mice were treated identically as in Experiment 1, except that the treated group received once-weekly intraperitoneal injections of 10 mg/kg gemcitabine. In contrast to Experiment 1, the combined treatment significantly reduced the relative tumor areas in the lungs. This result also resembles that of a phase III clinical trial (ORIENT-12), showing that patients with non-small-cell lung carcinoma benefited from combination treatment with gemcitabine and the anti-human PD-1 antibody sintilimab. Thus, this mouse model could be a feasible means to preclinically evaluate the antitumor efficacy of different immunotherapy and chemotherapy drug combinations.     

Enhancement of hepatocellular proliferative activity of kojic acid in mice by a simultaneous administration of ascorbic acid

To examine the tumor modification activity of kojic acid (KA) by sodium ascorbic acid (AA), 5-week-old male ICR mice were administered intraperitoneally with N-diethylnitrosamine (DEN) as an initiation treatment. Two weeks after the initiation treatment, animals were fed basal diet containing 0 (Group 1: DEN alone) or 3% KA (Group 3: DEN+KA), drinking water containing 5,000 ppm AA (Group 2: DEN+AA) or 3% KA and 5,000 ppm AA (Group 4: DEN+KA+AA) for 6 weeks. One week after the administration of KA and/or AA, all mice were subjected to two-thirds partial hepatectomy. At the end of the experimental period, all surviving mice were sacrificed and removed the liver. The liver weights of the Groups 3 and 4 were significantly increased, and the number of proliferating cell nuclear antigen positive hepatocytes and the gene expressions of Ccnc, Ccnd1, Ercc and Cyp7a1 were significantly increased in the Group 4, as compared to the Group 1. These results of the present study suggest that AA enhances the hepatocellular proliferative activity of KA in mice.     

谷物膳食纤维对高胆固醇饮食小鼠脂质代谢及肠道菌群的影响

本研究旨在探讨谷物膳食纤维(DF)对高胆固醇饮食小鼠的脂质代谢和肠道菌群的影响。选取C57BL/6J小鼠30只,并根据体重随机分为4组,分别饲喂基础饲粮(对照,CO组,n=8)、基础饲粮+5 g/kg DF(CO+DF组,n=8)、高脂高胆固醇饲粮(HF组,n=7)及高脂高胆固醇饲粮+5 g/kg DF(HF+DF组,n=7)。试验期8周,测定各组小鼠生长性能、血清和肝脏生化指标、粪便总胆固醇(TC)、短链脂肪酸(SCFA)、相关肝脏胆固醇和胆汁酸(BA)蛋白表达及肠道菌群。结果表明:1)CO+DF组的最终体重显著低于CO组(P<0.05),与HF组相比,CO组和HF+DF组的4周和最终体重均显著降低(P<0.05),肝脏和脂肪的重量显著减少(P<0.05)。且与HF组相比,DF+HF组摄食效率显著降低(P<0.05)。2)CO+DF组血清TC、甘油三酯(TG)含量及天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)活性以及肝脏TC和TG含量显著低于CO组(P<0.05),CO组和HF+DF组显著低于HF组(P<0.05),粪便TC含量仅在HF+DF组显著低于HF组(P<0.05)。3)DF对肝脏胆固醇调节元件结合蛋白-2(SREBP-2)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)、肝脏X受体α(LXRα)、微管相关蛋白1轻链3-Ⅰ(LC3-Ⅰ)和微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)蛋白表达没有显著影响(P>0.05)。CO+DF组肝脏胆固醇7-羟化酶(Cyp7a1)的蛋白表达水平显着低于CO组(P<0.05),但HF组和HF+DF组之间没有显著差异(P>0.05)。4)HF+DF组的粪便总SCFA和丙酸盐含量显著高于HF组(P<0.05)。与相应的对照组相比,CO+DF组和HF+DF组的粪便丁酸盐含量显著增加(P<0.05)。5)在门水平上,DF组厚壁菌门(Firmicutes)相对丰度减少,HF+DF组的拟杆菌门(Bacteroidetes)相对丰度高于HF组。梭状芽孢杆菌属ⅪⅤa(ClostridiumⅪⅤ)相对丰度在CO+DF组和HF+DF组显著增加(P<0.05)。此外,与其他组相比,仅HF+DF组中阿克曼菌属(Akkermansia)相对丰度显著增加(P<0.05)。由此可见,每千克饲粮添加5 g DF可降低饲喂含脂45%饲粮小鼠的体重和肝脏脂肪含量,降低血清TC、TG含量及AST和ALT活性以及肝脏TC和TG含量,降低胆固醇合成相关蛋白Cyp7a1的表达水平,提高总SCFA含量及肠道有益菌群和优势菌群相对丰度。     

兔IL-6基因真核表达载体的构建及其对pcDNA-VP60DNA疫苗佐剂效应研究

为研究兔白介素6（interleukin6,IL-6）真核表达质粒（pcDNA-IL-6）对核酸疫苗免疫效果的影响,本文构建了真核表达质粒pcDNA-IL-6,并将其与核酸疫苗pcDNA-VP60联合免疫家兔,以pcDNA-VP60和质粒载体pcDNA3．1（＋）作对照,用血凝抑制试验检测试验兔体内特异性抗体水平。结果表明：真核重组质粒pcDNA-IL-6对重组质粒pcDNA-VP60均具有免疫增强作用,从免疫后7d到70d,pcDNA-VP60／pcDNA-IL6联合免疫组抗体水平均高于pcDNA-VP60免疫组,差异具有显著统计学意义。     

The immunohistochemical evaluation of estrogen receptor-alpha and progesterone receptors of normal, hyperplastic, and neoplastic endometrium in 88 pet rabbits

The expression of estrogen receptor-alpha (ER) and progesterone receptor (PR) in normal, hyperplastic, and neoplastic endometrium in rabbits was evaluated by immunohistochemistry. The tissues evaluated were 27 normal uteri, 19 cases with endometrial hyperplasia, and 42 adenocarcinomas. Sixteen of 27 cases of normal uteri (59.3%) and 13 out of 19 hyperplasias (68.4%) stained positive with both ER-alpha and PR. Adenocarcinomas were further subdivided into 26 papillary and 16 tubular/solid adenocarcinomas. Papillary adenocarcinoma infiltrated the myometrium late in the disease and caused attenuation of the myometrium. In contrast, tubular/solid adenocarcinoma invaded into the deep myometrium early in the disease without thinning of the myometrium. Twenty-one cases out of 26 (80.8%) cases of papillary adenocarcinoma were both ER-alpha and PR negative, whereas 15 out of 16 (93.8%) of the tubular/solid adenocarcinomas were positive for ER-alpha, PR, or both. The total immunoreactive scores of ER-alpha, PR, and mode of myometrial invasion were significantly different between histopathologic types. This suggests that there may be 2 different developmental pathways for uterine adenocarcinomas in the rabbit.