催情助孕液对小鼠雌、孕激素水平及其受体mRNA表达的影响

分别以0.1、0.2、0.4 mL/d剂量的催情助孕液给21日龄小鼠灌服7d后,检测体质量、脏器指数、血清雌激素和孕激素水平、子宫组织中雌激素和孕激素受体基因mRNA的表达情况,并与腹腔注射雌二醇4d的小鼠进行比较.结果显示与对照组和雌二醇注射组比较,0.2 mL的催情助孕液能显著促进小鼠子宫发育,并增强雌激素水平及其受体基因表达,而对孕激素水平及其受体基因表达无显著影响.结果表明,催情助孕液对动物生殖发育与促进发情具有较好的效果.     

传染性支气管炎病毒感染对蛋雏鸡输卵管雌激素受体及孕激素受体表达的影响

以不同传染性支气管炎病毒（肾型IBV-河北分离株与呼吸型IBV-M41）分别感染蛋雏鸡,通过测定输卵管蛋白分泌部及子宫雌激素受体（Estrogen receptor,ER）和孕激素受体（Progesterone receptor,PR）的表达水平,来探讨IBV早期感染（10d）对抑制蛋鸡输卵管发育的作用机制。结果显示,IBV感染降低子宫和蛋白分泌部ER mR-NA、PR mRNA的表达水平,即致子宫ER mRNA和PR mRNA的表达低于检测水平;蛋白分泌部ER mRNA和PR mRNA的表达水平显著降低（P〈0.01）,且M41的作用幅度较肾型IBV更为显著（P〈0.01）。结果表明,IBV可通过降低ER和PR的表达水平,抑制输卵管对雌激素的应答能力而影响输卵管的发育。     

Regulation by gonadal steroids of estrogen and progesterone receptors along the reproductive tract in female lambs.

The regulation of estrogen and progesterone receptor (ER, PR) expression by estradiol (E2) and progesterone (P4) in the oviduct, uterus and cervix of female lambs was studied. The animals received three intramuscular injections of E2, P4 or vehicle with an interval of 24 h and they were slaugthered 24 h after the third injection. Determinations of ER and PR were performed by binding assays and mRNAs of ER alpha and PR by solution hybridization. High levels of ER and PR in both cervix and oviduct were found in the female lamb, differing from other mammalian species. No significant effects by either E2 or P4 treatment on ER and PR levels in the cervix and oviduct could be observed. E2 treatment increased the mRNA levels of ERa and PR more than 3-fold in the cervix, while P4 treatment increased the mRNA levels of ERa and PR in the uterus. The results show differential effects of gonadal steroids on sex steroid receptor expression along the reproductive tract in female lambs, suggesting that steroid target tissues can modulate responses to the same circulating levels of steroid hormones.     

Intestinal neoplasia in horses

BACKGROUND: Intestinal neoplasia of horses is inadequately described. HYPOTHESIS: Intestinal neoplasia of horses has characteristic clinicopathologic features. ANIMALS: Thirty-four horses with intestinal neoplasia. METHODS: Retrospective study. RESULTS: Anamnesis, clinical signs, clinicopathologic and pathologic findings in 34 adult horses diagnosed histologically with intestinal neoplasia were reviewed. The horses ranged in age from 2 to 30 years (mean 16.6 years at presentation). The Arabian breed was most represented and there was no sex predisposition. The most common presenting complaints were weight loss, colic, anorexia, and fever. The most consistent clinical signs were poor body condition, tachycardia, tachypnea, fever, and diarrhea. Useful diagnostic tools included rectal examination, routine blood analyses, abdominocentesis, ultrasonographic examination, rectal biopsy, and exploratory laparotomy. Alimentary lymphoma was the most common intestinal neoplasia identified, followed by adenocarcinoma and smooth muscle tumors. The small intestine was the most common segment of intestine affected for all neoplasms. Intestinal neoplasia was diagnosed antemortem in 13 of 34 (38%) horses. The median time from onset of clinical signs to death or euthanasia was 1.9 months. The discharge rate was 15%. Although the longest survival was observed in horses with jejunal adenocarcinoma, all horses were eventually euthanized because of intestinal neoplasia. CONCLUSIONS: Arabian horses were 4.5 times more likely to have intestinal neoplasia diagnosed than were other breeds.     

Immunohistochemical expression of progesterone receptors, growth hormone and insulin growth factor-I in feline fibroadenomatous change

The immunohistochemical expression, tissue-specific and cell-specific distribution patterns of progesterone receptors (PR), growth hormone (GH) and insulin growth factor-I (IGF-I) have been studied in 22 cases of feline fibroadenomatous change (FFAC). PR and GH were detected in all cases and were distributed homogeneously throughout the lesion, while IGF-I was detected in 77% of the cases at the site of ductal budding. The simultaneous expression of PR, GH and IGF-I was detected in epithelial cells in 14 of 22 cases while PR and GH expression only was detected in epithelial cells in 11 cases. Cases that expressed GH and IGF-I without PR expression in the stroma were the most numerous. Double immunohistochemical staining showed the co-localisation of PR and GH in a subset of ductal epithelial cells located between basal/myoepithelial and luminal cells (probably undifferentiated stem cells). These results suggest that ligand-activated progesterone receptors may induce the local synthesis of GH which in turn may exert its proliferative action directly and also indirectly through the production of other growth factors, such as IGF-I, in an autocrine/paracrine manner.     

大鼠不同时期乳腺组织中雌激素孕激素及催乳素的变化规律

为探讨雌激素(E2)、孕激素(P)及催乳素(Prl)在大鼠不同时期乳腺组织中的变化规律,选雌性SD大鼠42只,分别为处女鼠、妊娠鼠(6d、12d、18 d)、泌乳鼠(6d、12d、18 d),每个时期6只.在指定的时间点处死大鼠取乳腺组织.用放免(RIA)组织匀浆中E2、P及Prl水平进行定量研究,用统计学方法进行处理,观察这三种激素的变化规律.结果显示E2从处女期到整个妊娠期,其呈现下降趋势.而分娩后E2水平急剧升高,后又有所下降,但维持在较高水平;在乳腺发育不同时期,P在乳腺组织中整体水平一直呈现下降趋势;Prl水平以处女期最高;随着妊娠进行逐渐降低,整个乳腺发育过程中Prl整体上几乎一直呈下降趋势.表明E2、P及Prl在促进乳腺发育及维持和启动泌乳方面发挥了重要的作用.     

Clinical, histopathological and immunophenotypical findings in five horses with cutaneous malignant lymphoma

This study documents the clinical, histopathological, immunohistochemical and flow-cytometric findings in five horses with cutaneous non-epidermotropic malignant lymphoma (ML). The median survival time after discovery of the first subcutaneous nodules was 3.8 years (range 2-5 years: n=4). Histologically, the cutaneous ML had a pleiomorphic structure and contained a mixture of large reticulo-endothelial cells, medium-large sized lymphoid cells with a rounded nucleus and small nucleoli, many medium sized lymphoid cells with irregular nuclei, and some small lymphoid cells. Immunohistochemically (IHC) the lymphoid cells were positive for the pan-T-lymphocyte marker CD3 but negative for the B-lymphocyte markers CD21 and kappa and lambda immunoglobulin light chains. Although routine haematological examination revealed no abnormalities in the horses with cutaneous ML, changes in the peripheral blood lymphocyte population were apparent flow-cytometrically. Compared to clinically healthy horses, a decreased total percentage of cells was recorded in the lymphocyte gate. In three horses with cutaneous ML, an increase in CD4 positive cells was noticed in the monocyte gate. Flow-cytometric analysis of tumour cells collected by fine needle aspiration (FNA) suggested that the cutaneous MLs consisted primarily of CD4 and CD8 positive T-lymphocytes. The results were compared to those of a monomorphic multicentric T- and a monomorphic multicentric B-cell lymphoma. The results of immunohistochemistry and flow-cytometry were largely but not completely in accordance. In conclusion, the results of this study suggest that cutaneous non-epitheliotropic malignant lymphomas in the horse are of T-cell origin and that, after improvement of its accuracy, flow cytometric analysis of FNAs might become a useful aid to rapid tumour identification.     

Immunohistochemical localization of endometrial oestrogen and progesterone receptors in the cow

The presence and distribution of oestrogen and progesterone receptors have been investigated by means of immunohistochemical procedures in the uterus of two groups of cows: the first group underwent superovulatory hormonal treatment while the second group was used as a control group. After the immunohistochemical study no differences regarding the presence and distribution of hormone receptors seemed to be apparent between the two groups, so it was concluded that the topographical distribution and staining intensity of hormone receptors seem to be unaffected by hormonal superovulatory treatment.     

Bovine uterine, cervical and ovarian androgen receptor concentrations. Correlation with estrogen and progesterone receptor concentrations.

Bovine cytosol androgen receptor (ARC) concentrations were examined simultaneously in various regions of the uterus and in ovarian tissues of cows, and were related to cytosol estrogen (ERC) and progesterone receptor (PRC) concentrations and circulating steroid levels. ERC concentrations were 3-7-fold and PRC concentrations 13-29-fold those of ARC in bovine endometrial and myometrial tissues. When serum progesterone levels were low, both endometrial and myometrial ARC, endometrial ERC, and endometrial and myometrial PRC concentrations were higher (p < 0.05) than those observed during higher progesterone concentrations. Because serum 5 alpha-dihydrotestosterone (5 alpha-DHT) concentrations were higher during the luteal phase, it is possible that ARC was down-regulated by this natural ligand at this phase of the cycle. There were no differences between uterine horns in endometrial or myometrial ARC concentrations. Bovine cervical and ovarian stromal tissue also contained ARC, and the concentrations were about the same as in the endometrium and the myometrium. The relative binding affinities (RBAs) of some steroid hormones towards ARC in vitro were: the synthetic compound R1881 (146%), 5 alpha-dihydrotestosterone (100%), testosterone (75%) while estradiol-17 beta, progesterone and dexamethasone had lower RBAs (2, < 1, < 1% respectively). Cytosol androgen receptor concentrations correlated significantly with cytosol progesterone (PRC) and estrogen receptor (ERC) concentrations, both in the endometrium and myometrium. These data show that androgens, such as 5 alpha-DHT, may participate the endocrine regulation of bovine reproductive tissues.     

Expression of estrogen receptor 1 and progesterone receptor in primary goat mammary epithelial cells

The exact role and sensitivity of cells to estrogen and progesterone mediated through the steroid receptors during lactation is not known. Expression of estrogen receptor 1 (ESR1) and progesterone receptor (PGR) was quantified in mammary tissue‐derived primary goat mammary epithelial cells (pgMECs) to determine the influence of donor tissue physiology (lactating and juvenile) and cell culture growth conditions (basal and lactogenic) on ESR1 and PGR expression in the derived cells. Relative messenger RNA (mRNA) levels for both receptors were the highest in cell lines derived from mammary tissue of juvenile goats. Maintaining pgMECs in lactogenic conditions resulted in up‐regulation of ESR1 (1.36‐ to 12.35‐fold) and in down‐regulation of PGR (‐2.53‐ to ‐3.62‐fold), compared to basal conditions. Based on Western blotting analysis we suggest that the differences in mRNA expression are translated to the protein level. We suggest that differential expression in lactating conditions is correlated with terminal differentiation of the pgMECs. Double immunostainings showed that estrogen receptor alpha (ER‐α) positive cells do not exclusively belong to the luminal lineage and that ER‐α and PGR can be expressed individually or co‐expressed in the pgMECs. The derived primary cultures/lines in early passages are hormone‐responsive and represent a useful surrogate for mammary tissue in research experiments.     

Immunohistochemical assay for detecting estrogen receptors in canine mammary tumors.

OBJECTIVE: To determine the estrogen receptor (ER) content of canine mammary gland tumors by use of immunohistochemical (IHC) examination of formalin-fixed sections. SAMPLE POPULATION: 21 mammary gland tumors from 20 adult dogs. PROCEDURE: ER were detected in formalin-fixed tissues, using an avidin-biotin alkaline phosphatase IHC assay and were quantified on fresh-frozen tumor samples, using a modified dextran-coated charcoal (DCC) assay. RESULTS: 7 of 21 tumors had visually detectable nuclear ER by use of IHC staining, whereas 8 of 21 tumors were positive for ER by use of the DCC assay. The ER-positive cells in 5 IHC-positive tumors were epithelial cells with histologic criteria of early malignancy. The remaining 2 ER-positive tumors detected by use of IHC had ER-positive mast cells within areas of connective tissue around the tumor. CONCLUSIONS: Immunohistochemistry is an additional method for detection of ER in canine mammary tumors. The major advantage of this type of assay is that it may be performed on formalin-fixed tissues, and individual ER-positive cells may be identified. Discovery of ER-positive mast cells by use of IHC is of concern, particularly if the ER status of a tumor is based on DCC results alone. CLINICAL RELEVANCE: Because most canine mammary tumors are fixed in formalin prior to histologic evaluation, an IHC assay that identifies ER-positive cells is desirable. Adjunctive antiestrogen therapy could be administered to dogs with ER-positive tumors.     

Cytoplasmic estrogen and progesterone receptors in canine endometrium during the estrous cycle

Estradiol and progesterone receptors (ER, PR) were characterized and measured in cytosols from canine endometrium, using saturation and sucrose-gradient centrifugation radioassays. Both receptors were demonstrated to be steroid- and tissue-specific saturable proteins, which bound the respective steroids with high affinity (dissociation constant [Kd] approximately 10(-9)M). Serum estradiol, progesterone, and endometrial cytosol receptor concentrations and receptor-binding affinity were measured for 25 bitches from which samples were obtained at 5 stages of the estrous cycle (5 bitches each): anestrus (A), the 3rd day of proestrus (P3), the 3rd day of estrus (E3), the 12th day after onset of estrus (E12), and the 28th day after onset of estrus (E28). Mean (+/- SEM) serum estradiol concentrations were 17.0 +/- 2.2 (A), 55.4 +/- 5.0 (P3), 89.4 +/- 24.9 (E3), 41.0 +/- 5.9 (E12), and 50.6 +/- 3.9 (E28) pg/ml. Mean (+/- SEM) serum progesterone concentrations were 0.4 +/- 0.1 (A), 1.5 +/- 0.2 (P3), 17.3 +/- 7.5 (E3), 41.6 +/- 9.5 (E12), and 25.8 +/- 3.2 (E28) ng/ml. Concentrations of ER increased significantly from 1.06 pmol/g of uterus during stage A to a peak concentration of 6.18 pmol/g of uterus at E12, followed by a gradual decrease to 0.69 pmol/g of uterus by E28. The PR concentrations increased from 3.01 pmol/g of uterus in stage A to 17.32 pmol/g of uterus at P3; PR concentrations, thereafter, decreased gradually to 1.85 pmol/g of uterus by E28. Dissociation constants were significantly higher at E12 for the ER (Kd = 2.6645 X 10(-9)M) and at P3 for the PR (Kd = 5.8282 X 10(-9)M) than at the other stages examined, indicating a decrease in receptor affinity during the periods of high receptor concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Tamoxifen induces apoptosis and inhibits respiratory burst in equine neutrophils independently of estrogen receptors

Neutrophils play an important role in the exacerbation and maintenance of severe equine asthma; persistent neutrophil activity and delayed apoptosis can be harmful to surrounding tissues. Tamoxifen (TX) is a nonsteroidal estrogen receptor modulator with immunomodulatory effects and induces early apoptosis of blood and bronchoalveolar lavage neutrophils from horses with acute lung inflammation. This study investigated if the in vitro effects of tamoxifen are produced by its action on nuclear (α and β) and membrane (GPR30) estrogen receptors in healthy equine neutrophils. Results showed that TX inhibits neutrophil respiratory burst induced by opsonized zymosan in a dose‐dependent manner. Nuclear (17‐β‐Estradiol) and GPR30 cell membrane (G1) estrogen receptor agonists and their antagonists (ICI 182,780 and G15, respectively) do not block or reproduce the effect of TX. Therefore, TX does not inhibit respiratory burst through estrogen receptors. TX (8.5 μM) also increased phosphatidylserine translocation, a marker of early apoptosis, which did not occur with any of the estrogen receptor agonists or antagonists . Thus, tamoxifen generates dose‐dependent inhibition of respiratory burst and increased early apoptosis in healthy equine neutrophils, independently of nuclear or membrane estrogen receptors. Further studies are necessary to explore the signaling pathways of tamoxifen‐induced ROS inhibition and phosphatidylserine translocation.     

Morphologic, immunohistochemical, and molecular characterization of hepatosplenic T-cell lymphoma in a dog

A 13-year-old neutered male Jack Russell Terrier (Parson Russell Terrier) was presented to the Texas Veterinary Medical Center with a history of lethargy, depression, vomiting, and fever. The dog had mildly regenerative anemia, severe thrombocytopenia and low antithrombin activity. Marked splenomegaly was found on physical examination and imaging studies, and malignant round cell neoplasia and marked extramedullary hematopoiesis were diagnosed on aspirates of the spleen. The dog underwent exploratory laporatomy and splenectomy. Because of a rapid decline in clinical condition postsurgery, the dog was euthanized. Splenic and hepatic biopsies were submitted for histopathologic evaluation. A neoplastic population of round cells was found throughout the splenic parenchyma and within hepatic sinusoids. The neoplastic cells stained strongly positive for CD3 (T-cell marker) and were negative for CD79a (B-cell marker) and lysozyme (histiocytic marker). A diagnosis of T-cell lymphoma was confirmed by assessment of T-cell clonality using canine-specific polymerase chain reaction-based techniques. Although expression of the gammadelta T-cell receptor was not evaluated, this case shares many similarities with a rare syndrome in humans known as hepatosplenic gammadelta T-cell lymphoma.     

Metastatic melanoma in horses

The clinical and pathologic findings are reviewed for 14 horses with metastatic melanoma. All were older gray horses, with an average age of 16 years. The most common sites of primary tumors were the ventral tail, perineum, and parotid salivary gland. Metastases were found in multiple locations and caused a variety of clinical syndromes. The most common sites for metastases were the lymph nodes, liver, spleen, skeletal muscle, lungs, and surrounding or within blood vessels throughout the body. Many of the horses had dermal melanomas for years (range 1-6 years) before succumbing to metastatic disease. Histologic characteristic of dermal masses was not predictive of malignancy in the majority of cases. Treatment consisting of surgical debulking and administration of cimetidine, an autogenous vaccine, or both was attempted in 4 horses with no effect on outcome.     

Endocrinal responses in exercising horses

The intensity and duration of exercise exert a major influence on energy expenditure and physiological changes in the horse. Stressful environmental conditions, acclimation, and training status may further modify these responses. To maintain functional homeostasis during exercise, changes in autonomic nervous activity and hormone secretion are coupled to both the feedforward and the feedback mechanisms that control substrate mobilisation and utilisation.During exercise, both the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis are activated, which increases the circulating levels of adrenocorticotropin (ACTH), cortisol, adrenaline and noradrenaline. Furthermore, adrenaline inhibits the release of insulin from the pancreas. Catecholamines, adrenaline, and noradrenaline increase glycogen breakdown in the muscles. In the liver, catecholamines, together with cortisol, increase blood glucose by activating glycogen breakdown and gluconeogenesis. Cortisol and catecholamines also enhance the mobilisation of free fatty acids from fat stores.In addition to efficient energy metabolism, the ability to exercise is highly dependent on the well-coordinated neuroendocrine control of cardiovascular function. Catecholamines increase oxygen delivery during exercise by enhancing cardiac output, splenic erythrocyte release, and skeletal muscle flow. Furthermore, cardiovascular homeostasis is maintained by changes in plasma renin activity and in plasma concentrations of atrial natriuretic peptide (ANP), arginine vasopressin, and aldosterone.     

Validation of thromboelastometry in horses

Background: Thromboelastometry is used for identifying or monitoring coagulation abnormalities. It has been validated in several species but not in horses and the characteristics of the equine thromboelastogram have not yet been detailed. Objectives: The purpose of this study was to validate a thromboelastometer to be used with equine blood and to define the normal equine thromboelastogram. Methods: A Rotem-gamma thromboelastometer (Pentapharm GmbH, Munich, Germany) was used on 38 citrated blood samples to investigate native coagulation, the intrinsic and extrinsic pathways, the function of fibrinogen (largely dependent on its concentration), and the presence of fibrinolysis. Using classic validation approaches, we evaluated the imprecision of the method and the influence of hemolysis and storage time and temperature. The normal thromboelastogram was defined in both saddle and racing horses (the latter sampled before and after the race). Results: For imprecision tests, the analytical variations were <10%. The equine thromboelastogram had a pattern similar to those of other species, but the intrinsic and extrinsic pathways were less and more efficient, respectively. Reference intervals in racing horses, especially after exercise, were different from those of saddle horses, most likely due to a higher RBC mass. Coagulability decreased in hemolyzed samples and significant changes were found between nonrefrigerated and refrigerated blood samples stored for 20 hours. Conclusions: The Rotem-gamma thromboelastometer is a precise instrument for use with equine blood samples. The equine thromboelastogram is similar to that of other species, but reference intervals vary with aptitude and exercise. Hemolysis and refrigeration alter thromboelastometric results.     

Exercise-induced physiological adjustments to stressful conditions in sports horses

Among athletic/sports animals, the horse has a unique ability to increase its oxygen uptake by a factor of 60 during heavy exercise. This is achieved by physiological adaptations of all the links in the oxygen chain. Ventilation is increased by a factor of 30. Since the horse is a compulsory nasal breather, this hyperpnea necessitates high transmural pressure changes, which may be responsible for the dynamic collapse of the airways. Blood flow is increased by a factor of 10. Since the left ventricle is not very compliant, this increase necessitates a high filling pressure in the pulmonary circulation, which may induce capillary stress failure and exercise-induced pulmonary haemorrhage. Lastly, oxygen transport is improved by splenic contraction which increases haemoglobinemia by 50%.Sports horses frequently suffer from several problems, which are related either to endogenous or exogenous stresses experienced during their career. These stresses, caused by the use of the horse as a competition animal, may lead to several medical problems.At a systemic level, endogenous stresses include hyperkaliemia, lactacidemia, and hyperthermia; oxidative stress may induce problems at a general, and/or a pulmonary level.External factors, e.g. poor quality of inspired air, transport, hot and humid ambient conditions, and microbiological agents, may also induce abnormal body attacks, and lead to health problems.     

MCT1, MCT4 and CD147 gene polymorphisms in healthy horses and horses with myopathy

Polymorphisms in human lactate transporter proteins (monocarboxylate transporters; MCTs), especially the MCT1 isoform, can affect lactate transport activity and cause signs of exercise-induced myopathy. Muscles express MCT1, MCT4 and CD147, an ancillary protein, indispensable for the activity of MCT1 and MCT4. We sequenced the coding sequence (cDNA) of horse MCT4 for the first time and examined polymorphisms in the cDNA of MCT1, MCT4 and CD147 of 16 healthy horses. To study whether signs of myopathy are linked to the polymorphisms, biopsy samples were taken from 26 horses with exercise-induced recurrent myopathy. Two polymorphisms that cause a change in amino acid sequence were found in MCT1 (Val₄₃₂Ile and Lys₄₅₇Gln) and one in CD147 (Met₁₂₅Val). All polymorphisms in MCT4 were silent. Mutations in MCT1 or CD147 in equine muscle were not associated with myopathy. In the future, a functional study design is needed to evaluate the physiological role of the polymorphisms found.