Crb1在小鼠生后不同发育阶段子宫组织中的表达

应用HE染色方法和免疫荧光组织化学技术对小鼠生后不同发育阶段子宫组织结构的发育以及极性调控蛋白Crb1的定位表达进行了研究,结果显示,随个体发育,子宫管腔及子宫腺腔不断扩大,子宫内膜上皮持续增厚、固有层内子宫腺逐渐发达;4周龄时子宫腔面出现纤毛,随发育进程越来越发达;在各发育阶段的小鼠子宫组织中均有Crb1的表达,随个体发育表达呈现先增强后减弱的趋势,在8周龄时达到最强;Crb1主要定位于子宫内膜上皮细胞和子宫腺上皮细胞的胞膜和胞质,提示Crb1可能与小鼠子宫内膜上皮细胞和腺上皮细胞的极性建立和维持有关。     

小鼠生后不同发育阶段附睾组织中Crb3的表达

应用HE染色法和免疫荧光组织化学技术分别对小鼠生后不同发育阶段附睾组织结构及Crb3在不同发育时期附睾组织中的定位表达进行了研究。结果显示,附睾在4周龄时上皮为2层～3层的假复层纤毛柱状细胞,顶端纤毛细长;6周龄上皮细胞层数减少至1层～2层;8周龄附睾管上皮厚度增至最大,上皮细胞呈单层高柱状;12周龄后附睾管上皮开始变薄,呈单层柱状。免疫荧光染色结果显示,Crb3蛋白主要分布在附睾上皮柱状细胞的胞膜,在精子尾部有微弱的表达,这提示了Crb3不仅与附睾上皮细胞的极性建立和维持有关,而且对精子活力和血-附睾屏障的形成也可能具有潜在的影响。     

Fibrinous pericarditis and cardiac tamponade in a 3‐week‐old pony foal

A 3‐week‐old Pony of the Americas foal presented with a history of respiratory distress presumed to result from Actinobacillus equuli pleuropneumonia and septic arthritis. Failure of transfer of passive colostral immunity was suspected, but not confirmed, based on a history of the foal being separated from its dam shortly after parturition. Transient improvement was noted following thoracocentesis and removal of approximately 600 ml of pleural fluid but progressive clinical signs of congestive heart failure developed. Fibrinous pericarditis with evidence of cardiac tamponade was subsequently diagnosed via thoracic ultrasonography. Early clinical signs of cardiogenic shock were identified and fibrinopurulent exudate removed through a catheter placed with ultrasound guidance into the pericardium. The foal experienced cardiorespiratory arrest during the procedure and died despite resuscitative efforts. Post mortem examination identified extensive hypertrophy of the pericardium, septic arthritis, mild pleural effusion and focal bronchopneumonia. This report details the clinical evaluation, haematology, treatment and post mortem pathology of a foal with Actinobacillus equuli associated fibrinous pericarditis, as well as a brief review of cardiac tamponade.     

肌肉生长抑制素在小鼠不同发育阶段心肌组织中的表达

本试验旨在探究肌肉生长抑制素（myostatin,MSTN）基因在小鼠不同发育阶段心肌组织中的表达情况,分别选取幼年（7日龄）、性成熟（21~28日龄）、体成熟（42~56日龄）和老年（84日龄以上）4个发育阶段的健康小鼠（各3只）为研究对象,采用实时荧光定量PCR、HE染色、免疫组织化学等方法研究小鼠不同发育时期MSTN基因在心肌组织中的表达及不同时期心肌组织的发育。实时荧光定量PCR结果显示,MSTN基因在小鼠不同发育阶段心肌组织中均有表达,且其在小鼠老年时期的表达量最高,体成熟时期的表达量最低;幼年小鼠和性成熟小鼠心肌组织中MSTN基因表达水平显著低于老年小鼠（P<0.05）,而体成熟小鼠则极显著性低于老年小鼠（P<0.01）。HE染色结果显示,小鼠心肌细胞核及肌纤维的生长在不同发育时期明显不同,其中幼年时期的细胞核最多,肌纤维排列相对较紧密,而在性成熟、体成熟、老年时期心肌的发育指标依次降低。免疫组织化学结果显示,MSTN基因主要在心肌细胞核中表达,其各个时期表达量与实时荧光定量PCR的结果相符合。     

铁蛋白1基因在边缘革蜱各发育龄期、吸血期及不同器官中的表达分析

试验旨在了解铁蛋白1（ferritin 1,Fer1）基因在边缘革蜱各发育龄期、吸血期及不同器官中的表达情况。用实时定量荧光PCR法分析了Fer1基因在边缘革蜱各发育阶段、吸血期和不同器官中的相对表达情况,以饥饿雌蜱为基准（1 FC）,用2^-△△Ct法计算结果。结果显示,雌性蜱在开始吸血后第72 h Fer1基因的相对表达量最高,在刚刚孵化的饥饿成蜱（雌、雄）、卵及幼蜱中相对表达量较低;在若蜱中相对表达量较高,其中,幼蜱和若蜱在饥饿状态下比同时期饱血蜱的相对表达量高。在雌蜱的不同器官中,以饱血蜱唾液腺为基准（1 FC）,半饱血蜱中肠的表达量最高,在卵巢和马氏管不表达;在饱血蜱唾液腺、中肠、卵巢和马氏管中均表达,其中,在卵巢和马氏管中的表达量较高,中肠次之,唾液腺中的表达较低。试验结果表明,边缘革蜱体内Fer1基因的相对表达量随着蜱吸血过程变化而变化,这与蜱体内消化细胞的完善及消化酶在该过程中的变化相关,可为该类蜱铁代谢通路中关键功能基因的研究提供参考。     

Kiss-1基因在不同发育阶段五指山猪睾丸中的表达及定位研究

试验采用荧光定量技术研究亲吻素-1(Kiss-1)基因mRNA在30、60、80(初情期)和120日龄五指山公猪睾丸中的表达情况,并采用免疫组化技术定位其表达。结果显示,Kiss-1mRNA在五指山猪睾丸中的表达随着年龄的增长表达量逐渐升高,到初情期(80日龄)时达到最高(P0.05),随后到120日龄时显著降低(P0.05);免疫组化结果显示,睾丸间质细胞、精原细胞、初级精母细胞、次级精母细胞、圆形精子细胞及间质细胞中均发现Kiss-1阳性反应产物,而在精子中未发现Kiss-1阳性表达产物。结果证实,Kiss-1基因在五指山猪精子发生过程中发挥着重要作用。     

Mitral valve and tricuspid valve dysplasia in a 9‐week‐old Standardbred colt

A 9‐week‐old Standardbred colt was presented for investigation of dull demeanour, exercise intolerance and heart murmurs. Cardiac auscultation revealed a grade 5/6 holosystolic murmur and a grade 5/6 pansystolic murmur over the left and right cardiac apex respectively, and an irregularly irregular cardiac rhythm. Electrocardiographic examination findings were consistent with atrial fibrillation and tachycardia. Echocardiographic examination identified marked atrioventricular regurgitation and atrial dilation bilaterally, thickening of the mitral and tricuspid valves and dilation of the pulmonary artery consistent with pulmonary hypertension. No ventricular or atrial septal defect was present. Cardiomegaly and diffuse pulmonary oedema were evident on examination of lateral thoracic radiographs. Dysplasia of the mitral and tricuspid valves, eccentric cardiomegaly and pulmonary oedema were confirmed by post mortem examination. Dysplasia of the atrioventricular valves represents a rare cause of biventricular failure in the horse.     

不同发育阶段圭山山羊胚胎常规冷冻保存研究

采用常规冷冻法冷冻保存圭山山羊桑椹胚、囊胚、扩张囊胚期胚胎,解冻后体外发育率分别为41.94%、67.50%、84.09%,桑椹胚与囊胚、扩张囊胚间差异显著(P<0.05),囊胚和扩张囊胚间虽无显著性差异(P>0.05),但扩张囊胚体外发育率高于囊胚.在胚胎的冷冻-解冻过程中,桑椹胚、囊胚、扩张囊胚的透明带受损伤率分别为6.45%、20.00%、31.82%,透明带受损的桑椹胚、囊胚、扩张囊胚体外发育率分别为0%、50.00%、78.57%.初步说明发育程度越高的胚胎,在冷冻-解冻过程中其透明带越易受到损伤,但随发育程度的加深,胚胎生长发育时对透明带的依赖程度也变得越来越弱,表明圭山山羊早期扩张囊胚最适宜进行冷冻保存.     

雌性牦牛主要生殖器官中DBI的表达与定位分析

为加深对DBI基因功能的探究,揭示其在牦牛生殖生理过程中的作用.本试验采集健康母牦牛(3～6岁)繁殖周期不同阶段(卵泡期、黄体期和妊娠期)的卵巢、输卵管和子宫组织,共分为9组,每组设置3个生物学重复.采用实时荧光定量PCR (qRT-PCR)和蛋白免疫印迹技术(Western blotting,WB)检测牦牛DBI在繁...     

Expression of 11β‐Hydroxysteroid Dehydrogenase Type 1 and Glucocorticoid Receptors in Reproductive Tissue of Male Horses at Different Stages of Sexual Maturity

Glucocorticoids (GCs) as mediators of the stress response may affect Leydig cell function by inhibiting either luteinizing hormone receptor expression or testosterone biosynthesis. The isozymes 11β‐hydroxysteroid dehydrogenase (11βHSD) 1 and 11βHSD2 control the intracellular cortisol levels. Little is known about the effects of stress on fertility in the equine. The objective of the present study was to determine the presence and cellular localization of glucocorticoid receptors (GCR) and glucocorticoid‐metabolizing enzymes (11βHSD1 and 11βHSD2) in equine epididymal and testicular tissue with special regard to sexual maturation. Testicular and epididymal tissue was collected from 21 healthy stallions, and four age groups were designed: pre‐pubertal, young, mature and older horses. Immunohistochemistry (IHC) analysis and quantitative real‐time PCR (qRT‐PCR) were used. Pre‐pubertal horses showed higher testicular gene expression of 11βHSD1, 11βHSD2 and GCR than horses of all other groups (p < 0.05). A positive intranuclear immunoreaction for GCR was seen in epithelial cells of caput, corpus and cauda epididymidis and in Leydig cells. Significant differences (p < 0.05) between age groups occurred. The number of Leydig cells staining positive for GCR was highest in immature stallions (p < 0.05). The enzyme 11βHSD1 was localized in epithelial cells of the caput and corpus epididymidis and in Leydig cells. As determined by enzyme assay, nicotinamide adenine dinucleotide (NAD)‐dependant dehydrogenase (oxidation) activity was not detected in testicular tissue from immature stallions but in all other age groups (n = 3 per group). Results of this study suggest a contribution of GCs to maturation of male reproductive tissue in horses. In mature stallions, expression of 11βHSD enzymes and the oxidative 11βHSD activity in Leydig cells and epididymal basal and principal cells suggest a protective role on these tissues contributing to physiological intracellular glucocorticoid concentrations.     

Use of multi‐detector computed tomographic angiography in the diagnosis of a parapharyngeal aneurysm in a 6‐week‐old foal

Multidetector‐row computed tomographic contrast angiography (MDCTA) is routinely employed to investigate vascular masses in human patients but, to date, the use of this technique to investigate an aneurysmal mass has not been reported in an equine case. The potential of MDCTA to investigate a right‐sided parapharyngeal mass in a 6‐week‐old Thoroughbred foal was therefore investigated. A 4‐slice helical computed tomography scanner was used on a superficial, firm, ovoid mass yielding arterial blood on fine needle aspiration. MDCTA enabled identification of the vessels involved thus assisting in forming a diagnosis based on the morphology of the lesion and aided surgical planning. Histology confirmed the presence of mural thrombus and calcification within the smooth muscle wall consistent with a true aneurysm. MDCTA provided additional information to conventional imaging in this case of vascular masses of the head and neck. The technique therefore has the potential to improve diagnosis and assist in the management of such lesions.     

WIP1基因对3T3-L1前脂肪细胞增殖分化的影响及其在小鼠不同生长阶段的表达

【目的】探究野生型p53诱导磷酸酶1(WIP1)基因与脂肪细胞增殖和分化的关系,以期为猪优质性状育种提供新基因素材。【方法】利用脂质体转染方法将合成的WIP1基因的3对siRNAs (WIP1-790、WIP1-893和WIP1-1845)和阴性对照NC-siRNA分别转染3T3-L1前脂肪细胞,通过实时荧光定量PCR检测WIP1基因的表达水平,比较不同siRNA的干扰效率。然后利用筛选到的干扰效率最高的siRNA敲减WIP1基因在3T3-L1前脂肪细胞中的表达,通过CCK-8检测siRNA敲减细胞和NC-siRNA细胞不同生长时期(0、12、24、48、72、96和120 h)的增殖情况,并通过实时荧光定量PCR检测上述2种细胞24和48 h的细胞周期蛋白B1(Cyclin B1)和Cyclin D1的表达水平。对干扰效率最高的siRNA和NC-siRNA组3T3-L1前脂肪细胞进行成脂诱导分化,通过油红O染色和甘油三酯定量法评估成脂分化效率,利用实时荧光定量PCR检测过氧化物酶体增殖物活化受体γ(PPARγ)、CCAAT/增强子结合蛋白α(C/EBPα)、脂肪酸结合蛋白4(FABP4)、硬脂酰辅酶A去饱和酶(SCD1)的表达水平,Western blotting法检测PPARγ蛋白的表达水平;通过实时荧光定量PCR检测WIP1基因在21日龄、8周龄和6月龄小鼠腹股沟皮下脂肪组织和性腺脂肪组织的时空表达情况。【结果】基因干扰试验结果显示,3对siRNAs对WIP1基因的表达均具有极显著的干扰作用(P＜0.01),其中WIP1-790的干扰效率最高,达到了70%以上。CCK-8检测结果显示,与NC-siRNA组相比,WIP1-790干扰组细胞的增殖速率在各生长时期均极显著降低(P＜0.01),且细胞周期基因Cyclin B1和Cyclin D1的表达量在24和48 h均极显著下调(P＜0.01)。成脂诱导分化结果显示,与NC-siRNA组相比,WIP1-790干扰组细胞在成脂分化第8天油红O染色阳性细胞明显减少,脂滴含量极显著降低(P＜0.01),甘油三酯含量显著降低(P＜0.05);PPARγ、C/EBPα、FABP4、SCD1等标记基因mRNA表达水平均极显著降低(P＜0.01),PPARγ蛋白表达水平极显著降低(P＜0.01)。WIP1基因在8周龄和6月龄小鼠的腹股沟皮下脂肪组织中的表达量分别显著或极显著高于21日龄小鼠(P＜0.05;P＜0.01),且在6月龄小鼠的性腺脂肪组织中,WIP1基因的表达量极显著高于21日龄和8周龄的小鼠(P＜0.01)。【结论】WIP1基因通过调控Cyclin B1、Cyclin D1和PPARγ等细胞周期和成脂分化相关基因的表达影响3T3-L1细胞的增殖和分化,且参与小鼠脂肪沉积过程,结果可为深入解析WIP1基因调控脂肪发生的分子机制提供参考。     

Safety evaluation of orally administered afoxolaner and milbemycin oxime in eight‐week‐old dogs

The safety profile of afoxolaner (an isoxazoline molecule) when combined with milbemycin oxime (a macrocyclic lactone) was evaluated according to the regulatory requirements when administered six times orally in a soft chewable formulation at a dose of at least 1×, 3×, or 5× the maximum exposure dose in 8‐week‐old Beagle dogs. Thirty‐two healthy puppies (16 males and 16 females) were enrolled and allocated randomly to one of four treatment groups. Three doses were administered at 28‐day intervals (Days 0, 28, and 56), followed by three additional doses administered with 14‐day intervals (Days 84, 98, and 112). The study ended on Day 126. Treatment groups were as follows: Group 1: untreated, sham‐dosed control; Group 2: afoxolaner/milbemycin oxime chews administered at a dose of at least 5 and 1 mg/kg, respectively (1×); Group 3: afoxolaner/milbemycin oxime chews administered at a dose of at least 15 and 3 mg/kg, respectively (3); and Group 4: afoxolaner/milbemycin oxime chews administered at a dose of at least 25 and 5 mg/kg, respectively (5×). All dogs were examined for general health twice a day beginning on Day ‐14. Physical examinations, and blood collections for clinical pathology analysis and afoxolaner and milbemycin oxime plasma concentrations, were performed throughout the study. No afoxolaner/milbemycin oxime treatment‐related changes were observed in growth, physical variables, clinical pathology variables, or tissues examined histologically. No clinically relevant or statistically significant health abnormalities related to the administration of afoxolaner/milbemycin oxime were observed. No signs of macrocyclic lactone sensitivity were observed at any time during the study. Vomiting and diarrhea were observed sporadically across all groups including the controls. Based upon the results of this study, afoxolaner/milbemycin oxime soft chewables were shown to be safe when administered repeatedly at up to 5× the maximum exposure dose in dogs as young as 8 weeks of age.     

Successful treatment of infectious (Salmonella type III: 44) polyarthritis and osteomyelitis in a 4‐week‐old foal

This article describes the acute onset of infectious polyarthritis and osteomyelitis in a 4‐week‐old foal. Analysis of synovial fluid obtained from the left femoropatellar and right tarsocrural joints combined with clinical signs consisting of joint effusion and lameness yielded a diagnosis of septic arthritis. Bacterial culture of synovial fluid from the left stifle revealed Salmonella type III: 44. Rapid, sustained clinical improvement was noted following discontinuation of empirical antimicrobial therapy (potassium penicillin and amikacin sulphate) and initiation of treatment with ceftiofur and ampicillin. The importance of combining knowledge of veterinary pharmacology and microbiology so that appropriate antimicrobials may be selected with regard to the local environment in which they are to eradicate infection is emphasised. Despite frequent reference to amikacin sulphate as an effective antimicrobial for treating infections in foals caused by Salmonella, factors are discussed that explain why amikacin may not be clinically effective for treating infectious arthritis caused by Salmonella.     

Expression of VEGFR and PDGFR‐α/‐β in 187 canine nasal carcinomas

Radiotherapy represents the standard of care for intranasal carcinomas. Responses to tyrosine kinase inhibitors (TKIs) have been reported but data on expression of target receptor tyrosine kinases (rTKs) is limited. This study characterizes the expression of vascular endothelial growth factor receptor (VEGFR), platelet‐derived growth factor receptor (PDGFR)‐α and PDGFR‐β in canine intranasal carcinomas. Histological samples from 187 dogs were retrieved. Immunohistochemistry was performed using commercially available antibodies. Expression of rTKs was classified into weak, moderate or intense and additionally recorded as cytoplasmic, membranous, cytoplasmic‐membranous, nuclear or stromal. VEGFR was expressed in 158 dogs with predominantly moderate expression (36.9%) and a cytoplasmic‐membranous expression pattern (70.9%). PDGFR‐α was detected in 133 with predominantly weak expression (57.9%) and cytoplasmic pattern (87.9%). PDGFR‐β was identified in 74 patients with a predominantly moderate expression (17.6%) and cytoplasmic expression pattern (63.5%). Co‐expression of rTKs was common. These results confirm expression of VEGFR, PDGFR‐α and PDGFR‐β in canine intranasal carcinomas and support the utility of TKIs.