广西巴马小型猪Oct4基因克隆及其在成纤维细胞中的表达

根据GenBank中公布的猪Oct4核苷酸序列设计合成1对引物,采用RT-PCR方法从巴马小型猪睾丸组织扩增Oct4基因编码全序列。然后将该基因重组于含有绿色荧光报告基因的真核表达载体pEGFP-N1中,构建成pEGFP-Oct4重组质粒,通过酶切电泳鉴定和DNA测序证明重组质粒构建成功。使用脂质体2000将pEGFP-Oct4转染巴马小型猪肾脏成纤维细胞,荧光显微镜下可观察到绿色荧光,转染细胞株中可检测到Oct4的转录。结果表明,成功构建了真核表达载体pEGFP-Oct4,并可在巴马小型猪肾脏成纤维细胞中表达,为进一步研究Oct4基因生物学功能奠定了基础。     

不同代次广西巴马小型猪内源性反转录病毒3′UTR克隆及结构和调控元件分析

利用CDNA末端快速扩增技术（RACE）成功扩增了广西巴马小型猪近交系连续4代次及封闭群的猪内源性反转录病毒（PERV）3UTR,共获得8条序列（GenBank登录号分别为：GQ475493、GQ475494、GQ475495、GQ475496、GQ475497、GQ475498、GQ475499、GQ475500）,分为638、749bp 2种不同的类型,都属于PERV-A亚型。以PERV-3UTR构建遗传进化树,PERV-A,B,C分群清晰,3UTR可作为PERV亚型区分的另一基因序列。对调控元件定位分析,推测具有749bp类型3UTR的PERV病毒转录水平更高。在所获得序列的209～227bp位置,发现了＂CTTGAAACTT＂10个碱基的1.9个重复。模拟RNA二级结构,广西巴马小型猪PERV-3UTR 2种类型无论从空间构型、自由能,还是茎环数量都存在明显的不同。这些结果为全面评价广西巴马小型猪来源的PERV病原安全性提供了参考。     

A Comparison of the Histological Structure of the Placenta in Experimental Animals

The primary function of the placenta is to act as an interface between the dam and fetus. The anatomic structure of the chorioallantoic placenta in eutherian mammals varies between different animal species. The placental types in eutherian mammals are classified from various standpoints based on the gross shape, the histological structure of the materno-fetal interface, the type of materno-fetal interdigitation, etc. Particularly, the histological structure is generally considered one of the most useful and instructive classifications for functionally describing placental type. In this system, three main types are recognized according to the cell layers comprising the interhemal area: (1) epitheliochorial type (horses, pigs and ruminants), (2) endotheliochorial type (carnivores) and (3) hemochorial type (primates, rodents and rabbits). The number of cell layers in the interhemal area is considered to modify the transfer of nutrients between maternal and fetal blood and is one of the important factors with respect to the difference in placental permeability between animal species. Therefore, in reproductive and developmental toxicity studies, careful attention should be paid to the histological structure of the interhemal area when extrapolating information concerning placental transfer characteristics to different animal species.     

Bilateral cataract in a cynomolgus monkey

Severe bilateral cataract was found in a 7 year-old naïve female cynomolgus monkey (Macaca fascicularis) 3 months before necropsy. During macroscopic examination, severe opacity and thinning of the lens were observed in both eyes. Histopathology revealed that the lens nuclei and majority of cortex lens fibers had disappeared and become excavated, while the lens fibers in the subcapsular area were swollen and distorted. Other observations included atrophy and vacuolation in the lens epithelial cells and proliferation of spindle cells and collagen fiber beneath the anterior capsule of the right eye. Immunohistochemical staining of these spindle cells revealed the presence of vimentin, cytokeratin and α-smooth muscle actin (α-SMA), which were considered to be derived from lens epithelial cells. This is a rare case of spontaneous, bilateral, hypermature cataract in a cynomolgus monkey.     

Porencephaly in a cynomolgus monkey ( macaca fascicularis )

Porencephaly was observed in a female cynomolgus monkey (Macaca fascicularis) aged 5 years and 7 months. The cerebral hemisphere exhibited diffuse brownish excavation with partial defects of the full thickness of the hemispheric wall, and it constituted open channels between the lateral ventricular system and arachnoid space. In addition, the bilateral occipital lobe was slightly atrophied. Histopathologically, fibrous gliosis was spread out around the excavation area and its periphery. In the roof tissue over the cavity, small round cells were arranged in the laminae. They seemed to be neural or glial precursor cells because they were positive for Musashi 1 and negative for NeuN and GFAP. In the area of fibrous gliosis, hemosiderin or lipofuscin were deposited in the macrophages, and activated astroglias were observed extensively around the excavation area.     

Ocular fundus images with confocal scanning laser ophthalmoscopy in the dog, monkey and minipig

Confocal scanning laser ophthalmoscopy (CSLO) is a new technique that enables ocular fundus image recording and retinal dynamic angiography to be performed. The ocular fundus image is acquired sequentially, point by point, and is reconstructed on a video monitor at the rate of 25 images per second. The feasibility of performing both ocular fundus image recordings and retinal angiography image recordings were tested on two dogs, two monkeys and two minipigs using a 40° field I + Tech CSLO. Fundus area of each dog, monkey and minipig were examined without any additional optical devices. The ocular fundus and angiography images were recorded, stabilized and analyzed under the same conditions. For each species, all images were easily recorded without any additional optical device in a lighted room and the morphology of the retinal images generated was similar to those obtained with a camera or angiography of higher resolution. Capillary phase or venous times are presented. Image recording at 25 frames/second enabled more retinal dynamics to be demonstrated than with use of regular angiography. This technique is noninvasive and easy to perform if the eye is fixed and eyelids maintained open. It also allows exploration of retinal microvascularization and could be utilized for clinical, pharmacologic and toxicologic investigations as well.     

恒河猴辐射后的临床症状及护理

经5-7Gy的钴-60多向辐射后,恒河猴表现出食欲废绝,精神不振,活动性降低,呕吐,腹泻,脱水,易感染,易出血,最后死亡.这主要是因为辐射损伤了动物的造血机能和消化系统.临床上应对症护理,减轻动物疼痛.     

Goblet cell hyperplasia and muscular layer thickening in the small intestine of a cynomolgus monkey

We report here the interesting case of a 5-year-old male cynomolgus monkey with goblet cell hyperplasia and thickening of the muscular layer throughout the small intestine without exhibiting any clinical symptoms. Necropsy examination showed diffuse thickening of the intestinal wall from the jejunum to the ileum, with an appearance likened to a rubber tube. Histopathologically, marked thickening was observed in both the mucosal and muscular layers in the jejunum and ileum, and slight thickening was observed in the duodenum. Goblet cell hyperplasia with extension of the circular folds and villi was prominently observed. The mucosal surface was covered with a thick mucus layer containing desquamated mucosal epithelial cells, and both the inner and outer muscular layers were markedly thickened due to smooth muscle hypertrophy. Neither macroscopic nor histopathological examination identified any causative factors, such as infection, enteritis and intestinal stenosis, or obstruction that may have caused development of this lesion. Given these observations, this case may simply be considered of spontaneous goblet cell hyperplasia and muscular layer thickening in the small intestine of a cynomolgus monkey.     

Digestive retention times for Allen's swamp monkey and L'Hoest's monkey: data with implications for the evolution of cercopithecine digestive strategy

Primates access energy from plant fiber via bacterial fermentation in either a modified forestomach ('foregut'), a caecocolic ('hindgut') chamber of the large intestine, or both. Longer digestive retention times allow for more complete fermentation; as such, primates that consume an herbivorous diet high in fiber are expected to have both relatively and absolutely longer retention times than those mammals that rely on more readily digestible plant foods, such as fruit. We used particulate markers to measure the digestive retention times of captive Allen's swamp monkeys [Allenopithecus nigroviridis (Pocock, 1907)] (n= 3) and L'Hoest's monkey (Cercopithecus lhoesti P. Sclater, 1899) (n= 2). Results indicate mean retention times of 23.2-29.4 h and 23.2-24.0 h for C. lhoesti and A. nigroviridus, respectively. Results from this study, in combination with previously published data on digestive retention times in other primate species, indicate that cercopithecines differ from other primate taxa by having lengthier retention times that can be predicted by body mass alone. These data are consistent with the hypothesis that relatively lengthy retention times are a primitive trait for Cercopithecinae.     

Study of retinal microvessels in a Rhesus monkey model of chronic high intraocular pressure

Objective  To investigate the changes in retinal vessels, especially macular capillaries, under high IOP using the Rhesus monkey high IOP model.
Methods  The trabecular meshwork of the adult Rhesus monkey was cauterized by laser to induce increased IOPs with different degrees of damage. The eyeballs were enucleated, and the optic nerves were stained with toluidine blue in semithick slices. Part of the retina was observed under electron microscope, and the rest was stained by the ADPase method. The damage levels of the optic nerve were evaluated by axon count, and the pathological appearance of the macular capillaries were observed.
Results  Five mildly damaged eyes, three moderately damaged eyes and three severely damaged eyes were evaluated. Dense and intact perifoveal vascular rings were observed in all the eyes. The vessels' area percentages, as well as area, perimeter and diameter of the foveal avascular zones, were measured, and no statistically differences were found among different groups ( P -values were 0.269, 0.500, 0.951, and 0.555 separately). The ultra structures of the normal capillaries showed regular tubes and intact basement membranes, while lipoid substances in capillary tubes, swollen mitochondria in endothelial cell bodies, and uneven basement membranes were found in the high IOP-damaged eyes.
Conclusions  Compared with normal eyes, no obvious differences were found in macular microvessels and foveal avascular zones in the Rhesus monkey model of high IOP. However, presence of swollen mitochondria in endothelial cells and lipoid substances in capillary tubes might suggest that high IOP could damage the capillary endothelial cells.     

Comparative fluorescein angiography of the normal sheep and goat ocular fundi

Fluorescein angiography without sedative or anesthetic agents was evaluated in 20 normal goats and 20 normal sheep. All of the angiographic phases were observed using 20 mg/kg fluorescein IV in both species. Fundus fluorescein angiography results revealed wide stars of Winslow in the tapetal fundus, central or marginal flow during the first part of the arterial phase, delayed filling of the focal areas in the choroid near the optic disc that often coincided with others in the disc, and lack of evidence of the 'striate area' in the tapetal fundi. In goats, the angiographic times were 6.54+/-1.25 s for the arterial phase (TA), 7.80+/-1.37 s for the arterio-venous phase (TAV), and 14.13+/-2.01 s for the venous phase (TV). I1: 1.30+/-0.30 s (time elapsing between TA and TAV), and I2: 6.20+/-1.60 s (time elapsing between TAV and TV). In sheep, times were 9.54+/-2.18 s TA, 11.73+/-2.10 s TAV, and 20.86+/-2.74 s TV. I1: 2.04+/-0.75 s and I2: 8.98+/-2.47 s, respectively. Due to the large size of the fundic vessels in sheep and goats, fluorescein angiography of the retinal vasculature can facilitate the study of the different vascular diseases in these species.     

Non-selective computed tomography angiography of a vascular ring anomaly in a dog

Non-selective computed tomography angiography (CTA) is an emerging imaging technique that will have continued application in veterinary medicine because of its short scan time, use of a single, peripheral venous injection and availability of more detailed anatomic information—including 3-dimensional reconstructions. The improved anatomic detail may facilitate veterinary students' learning of complex aortic arch malformations and details relative to the surgical approach. Herein we describe application of its use in a dog with a vascular ring anomaly due to a persistent right fourth aortic arch and left ligamentum arteriosum. Additionally, we compare images with a normal dog.     

Comparison of cardiac measurements by multi-detector computed tomography angiography and transthoracic echocardiography in English bulldogs

Introduction

Transthoracic echocardiography (TTE) is the primary tool for the assessment of cardiac structure and function in dogs but is challenging in English bulldogs due to dorsoventral compression of the thorax, obesity, and narrow intercostal spaces. Multi-detector computed tomography angiography (CTA) may overcome the conformational obstacles of cardiac imaging in this breed.

Nonselective computed tomography angiography for detecting arterial blood flow to the distal limb following trauma in two small equids

Two equids weighing <250 kg were examined several days after suffering severe mid‐diaphyseal dorsal metatarsal lacerations. Distal limb vascular disruption was suspected in both cases. Nonselective computed tomography angiography with contrast medium injected peripherally via the jugular veins was used to evaluate the vascular supply to the distal limb. The use of this imaging technique demonstrated either intact or disrupted distal limb vasculature. The imaging results were verified with a positive long‐term outcome in the case with a diagnosis of an intact vasculature and with histological findings of avascular necrosis in the case with a diagnosis of disrupted vasculature.     

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.     

巴马小型猪对高致病性猪繁殖与呼吸综合征病毒的敏感性评价

为评价巴马小型猪对高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)的敏感性,本研究选择PRRSV抗体阴性的16头巴马小型猪和17头本地二元杂交猪作为研究对象,分别分为低剂量接毒组、高剂量接毒组、对照组1和对照组2,低剂量、高剂量接毒组肌肉注射接种PRRSV NVDC-JXA1强毒株,对照组1和接毒组混合饲养,对照组2分开饲养作为空白对照。接毒后每天测量体温,观察精神、食欲、死亡等情况,死亡动物剖检病变,攻毒后每隔7d采血用RT-PCR法检测病原,连续观察21d。结果绝大部分猪体温升高到41℃以上,且有3个以上温次,所有接种及混养动物都死亡,巴马小型猪死亡时间在8d~17d,二元杂交猪在7d~13d,死亡动物出现肺充血、出血、实变,扁桃体、淋巴结、肝脏、肾脏、皮下有出血等症状,病原RTPCR检测为阳性,说明攻毒动物死于HP-PRRS,表明巴马小型猪对PRRSV NVDC-JXA1强毒株非常敏感,可用于PRRS活疫苗的检验。     

Effects of three methods of restraint an in rhesus and african green monkeys intravenous glucose tolerance testing

The intravenous administration of 0.75 gm glucose per kg and the measurement of serum glucose pretest and at 10, 20, 30, 60, 90 and 120 minutes constitute a satisfactory protocol for intravenous glucose tolerance testing of Rhesus (Macaca mulatto) and African Green (Cercopithecus aethiops) monkeys. No significant differences were noted between animals restrained with ketamine hydrochloride and those restrained with sodium pentobarbital, but the African Green males and females and the male Rhesus monkeys yielded significantly different results while being manually restrained.