Neuroendocrine (NE) Cells in Rat Neonatal Lungs. A Histochemical and Immunocytochemical Study

A study has been carried out to determine the presence of NE cells in the newborn rat lung. The localization of these cell was achieved by an argyrophil method. Both single NE and NEB, were found. Immunoperoxidase techniques were performed to determine NSE, serotonin and calcitonin production in these NE formations.     

Chromogranin- and Somatostatin-containing Neuroendocrine Cells in the Porcine Uterus An Immunocytochemical Study

Endocrine cells scattered in organic mucosae were defined "Neuroendocrine" (NE) cells because they constitute a section of the Diffuse Neuroendocrine System (DNES). Such cells have never been evidentiated in the normal endometrium. By means of histochemical and immunohistochemical techniques, NE cells, some of which contain the hormone somatostatin, are described in the glandular epithelium of the uterine horn in non-pregnant, non-castrated, young and adult sows. As is known, the uterine horn is the organ of pregnancy in the pig. The localization, distribution and morphology of the uterine NE and somatostatin-containing cells are reported and the importance of their function, in the pregnant and non-pregnant porcine uterus, is discussed.     

弥散神经内分泌系统研究概论

本文系统概述了弥散神经内分泌系统(DNES)概念的形成、DNES的组成及其细胞结构、功能特点和DNES细胞的鉴定方法,并简要概述了DNES与疾病及免疫的关系。指出从免疫-神经-内分泌网络的角度来揭示DNES与消化道疾病及其黏膜免疫的关系将是研究胃肠道疾病发病机理的一个新思路。     

Fine Structural and Histochemical Study of Equine Paneth Cells

Ultrastructure. lysozyme and glycoconjugate activity in duodenal Paneth cells were observed concurrently in the horse. Paneth cells were seen to uniformly line the base of the equine intestinal glands. The round secretory granules have centrally located electron densities with peripherally located electron lucent halos. Histochemically, the peripheral halo layer was positively stained for carbohydrates by the periodic acid-thiocarbohydrazide-silver protein-physical development (PA-TCH-SP-PD) method and the entire granules reacted positively to the WGA. The central core area reacted with anti-lysozyme. We identified a young (Type I) and an old (Type II) cell population in the same crypt, but we suggest that the observed populations are variations of the same cell type with the varied appearance due to aging of the secretory granules.     

Neuroendocrine Cells in the Gastrointestinal Tract of Wild Boar

The presence and the distribution of neuroendocrine cells in the gastrointestinal tract of adult wild boar were investigated. The endocrine cells have been identified by means of immunocytochemical techniques using antibodies against serotonin, gastrin, somatostatin, cholecystokinin (CCK), met-enkephalin (MET-ENK), gastric-inhibitory peptide (GIP) and glucagon. The number of positive cells for each antiserum in each region was evaluated. Results were compared with data present in the literature and obtained previously by us and other authors in swine and domestic mammals (Ceccarelli et al., 1985, 1987, 1990, 1991, 1995; Capella and Solcia, 1972; Domeneghini and Castaldo, 1981; Peranzi and Lehy, 1984; Krause et al., 1985).     

Study on Inducing Equine Bone Marrow Mesenchymal Stem Cells Differentiated into Chondrocytes

This study was aimed to establish equine bone marrow mesenchymal stem cells(BMSCs) line and induce it to differentiate into chondrocytes. The BMSCs were collected by cutting the bone and flushing the cutting surface by PBS, and then the bone marrow was washed with PBS,the collected cells were cultured after centrifugation. The cells were purified by passaging,the stem cell properties were tested before the induction. And the cells were also appraised by the expression of the special gene of chondrocytes as well as staining the differentiated cells with Alcian blue to insure the induction was effective. The obtained BMSCs expressed Sox2 and Nanog genes, which were the stem cell special genes, and also expressed CD44, CD90 and CD105 genes, but absent of CD34 and CD45 genes. The shapes of the 3rd passage BMSCs were changed after cultured in inducing medium for a few days. Furthermore, the cells were positive to Alcian blue staining, increased expression of the Col special gene of chondrocyte day by day as well. According to this study, the BMSC line was established, and the BMSCs were induced and differentiate into chondrocytes successfully.     

马骨髓间充质干细胞向软骨细胞的诱导分化

试验旨在建立马骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）细胞系,并诱导其向软骨细胞分化。通过获取马BMSCs,进行细胞培养和纯化,对第3代（P3）细胞进行干细胞特性鉴定,并诱导其向软骨细胞分化,对分化后的细胞染色,并检测其软骨细胞特异性基因的表达。结果显示,获得的马BMSCs表达标记基因Sox2和Nanog,并表达间充质干细胞表面标记因子CD44、CD90和CD105,不表达造血细胞表面标志物CD34和CD45。P3代细胞经诱导培养后形态发生改变,阿尔新蓝染色为阳性,并表达软骨细胞特异基因Col,且其表达量随着诱导分化时间的增加而增高。综上表明,本试验建立了马BMSC细胞系,并成功诱导其分化为软骨细胞,为软骨损伤的干细胞治疗提供了试验依据。     

Primary Cilia in Chondrogenic Differentiation of Equine Bone Marrow Mesenchymal Stem Cells: Ultrastructural Study

Locomotor system disorders in equine species, such as tendon lesions, osteoarthritis, or ligament injuries, are some of the most frequent causes of dramatic reduction in horse performance. Traditional therapies are aimed at the inflammatory process and pain, but they do not regenerate normal tendon or ligament matrix and do not reduce re-injured rates. Mesenchymal stem cells have started to use as therapeutic option to repair these injured tissues. Most studies have focused on their isolation, in vitro culture and phenotyping. However, mesenchymal stem cell ultrastructure has been disregarded in the last years. We investigate the ultrastructural characteristics of these cells once differentiated into chondrocytes. Ultrastructural analysis was conducted on suspension cultures of differentiated chondrocytes from bone marrow mesenchymal stem cells by means of transmission electron microscopy. The morphologic characteristics of these cells, their ability to produce the extracellular matrix, and the presence of a single cilium could be indicative of the mesenchymal cells differentiation into chondrocyte phenotype. This study provides essential data to evaluate the degree of suitable phenotypic stability, for these cells can be used with repair purposes.     

Observations on the Equine Hypophysial Portal System

The blood supply to the equine pituitary gland, with special attention to its portal system, was studied by the aid of vascular injections and histological serial sections. The primary capillary plexus of the median eminence was supplied by the rostroventral and rostrodorsal infundibular arteries, which were branches of the internal carotid arteries and the ramus communicans caudalis, respectively. The neural lobe was supplied by the caudal infundibular arteries, which originated from the arteria intercarotica caudalis. Dorsal and ventral groups of the portal vessels were described. Their regional distribution into the sinusoidal capillaries of the pars distalis was discussed. Results of this investigation indicated that the portal vessels were the sole supply to the equine pars distalis. No direct arterial supply to this part of the adenohypophysis was observed.     

Immunocytochemical Study of the Ultimobranchial Tubule in Wistar Rats

A systematic immunohistochemical study of the ultimobranchial tubule (UBT) has been carried out in 45 Wistar rats of different ages (0, 5, 10, 15, 20, 25, 30, 60 and 120 days). The existence of calcitonin immunoreactive cells in the UBT wall has been demonstrated in a 5-days old rat. In addition, immunohistochemical studies for thyroglobulin revealed positive staining in follicular cells connected to the UBT and, occasionally, in isolated cells lying within solid clusters from the UBT. These last results together with the continued and repeated existence of numerous mitosis and PAS (+) microfollicles, apparently rising from the UBT, support the hypothesis that the ultimobranchial body (UBB) may contribute partially to the formation of a part of the follicular component.     

蒙古马胎儿成纤维细胞永生化细胞体系建立的研究

【目的】试验旨在建立永生化蒙古马胎儿成纤维细胞(equine fetal fibroblasts, EFFs)系,为马繁殖与发育生物学研究提供参考。【方法】采用1 mg/mL胶原酶Ⅳ消化分离原代EFFs,利用表达人端粒酶逆转录亚基(hTERT)和潮霉素B抗性的pLV-hTERT-IRES-hygro慢病毒感染EFFs,经潮霉素B筛选得到阳性细胞hTERT-EFFs,以第3代EFFs为对照,采用实时荧光定量PCR检测hTERT mRNA的表达水平,CCK-8法绘制生长曲线检测细胞增殖能力,流式细胞仪检测细胞凋亡情况,β-半乳糖苷酶染色检测细胞衰老情况。使用DNAMAN软件比对分析人、马、小鼠、牛、绵羊、猪多物种TERT核苷酸、蛋白和端粒酶RNA组分(TERC)序列相似性。【结果】原代EFFs在实验室培养体系下可传至17代,但第17代细胞发生了衰老。hTERT基因可成功转入EFFs,并至少稳定表达20代。然而hTERT-EFFs增殖能力显著低于EFFs(P<0.05),hTERT-EFFs凋亡早期细胞率显著高于EFFs(P<0.05)。细胞衰老检测结果表明,hTERT-EFFs...     

Equine Veterinarians' and Health Care Professionals' Concerns Related to the Implementation of the National Equine Identification System

The National Animal Identification System (NAIS) is an animal tracking database created to contain the spread of contagious foreign animal diseases. Currently, no studies address the perceived support of the equine veterinary field for NAIS. The objectives of this study were to determine equine veterinarians' level of familiarity with NAIS, perceived effectiveness of NAIS, use of equine microchipping, and extent of disaster planning. A 26-question online survey (Zoomerang zPro, MarketTools, Inc., San Francisco, CA) was designed and administered to a sample (n = 139) drawn from the American Association of Equine Practitioners. SPSS 16.0 (SPSS Inc., Chicago, IL) was used to analyze data. Results showed that only 19.4% of practicing veterinarians were “very familiar” with NAIS. Nearly half (49.3%) of the respondents were neutral regarding their opinion of NAIS. However, 55.6% thought that NAIS would efficiently stop the spread of a contagious animal disease. More than 52% reported that they were currently microchipping horses. After microchipping, 66.7% did not verify premises' identification numbers or record information about the horse, and 80.2% did not complete the paperwork to contact the State Department of Agriculture. Approximately 75.2% of respondents reported that they had experienced a contagious disease outbreak. Most (81.6%) veterinarian respondents did not have a plan to deal with clients' horses during a disaster. There is a need for communication and information for veterinarians regarding NAIS, premises registration, emergency planning guidelines, data to collect after microchipping, and how to provide information to clients.     

The Blood-Vessel System of the Equine Periodontal Ligament: a Mesoscopic Study on Corrosion Casts

Several investigators have suggested that the blood-vessel system in the equine periodontal ligament does not only serve nutritional needs, but also specific functional needs, e.g. in terms of a shock absorbing system. In order to supplement the previous data, corrosion casts of the periodontal blood vessels were studied with special attention to the spatial arrangement and connections of the vessels. The heads of eight warm-blooded horses (age 2.5–23 years, seven female, one male), which had been killed for medical reasons, were dissected. The arteria alveolaris inferior and the a. infraorbitalis were perfused with 100 ml Heparin (20 000 IU) followed by 100 ml tap water. After storage at 4°C for 24–96 h, 10–200 ml methacrylate resin (Biodur^TM E20) were injected by use of a cartridge pistol for approx. 1 h. After polymerization for 24 h at 60°C, the jaws were frozen and cut with a steel band saw in a horizontal, sagittal or transversal plane to obtain segments that exposed the periodontal space. The specimens were macerated in detergent at 60°C until the soft tissues had vanished. Care was taken to keep the periodontal vessels complete and in topographical integrity. The vascular casts were mesoscopically examined by use of a dissection microscope (magnification 5.8×–40×). The blood vessels in the periodontal space connected with vessels in three other sites:

• (1)

  occlusal – with the blood vessels of the gingiva;

       
  
  

二甲苯对小鼠肺组织损伤的初步研究

为了研究二甲苯对小鼠肺组织的损伤情况,以健康成年小鼠作为试验动物,分3个试验组(低、中、高剂量组)1个对照组,给试验组腹腔注射不同剂量二甲苯(0.125、0.25、0.5 mL/kg),给对照组腹腔注射生理盐水.15 d后剖检取材,采用石蜡切片和HE染色技术研究二甲苯染毒后小鼠肺组织结构的变化特点;采用分光光度法测定小...     

Oestradiol Induced Inhibition of Neuroendocrine Marker Expression in Leydig Cells of Adult Rats

The objectives of this work were to determine the changes in the expression of neuroendocrine markers in Leydig cell by oestradiol treatment, and to determine whether testosterone is able to recover partially the effects of hormonal suppression induced by oestradiol. Adult male rats were injected daily with either 50 microg of oestradiol or oestradiol plus testosterone propionate (25 mg every 3 days) for 15 days. The animals were sacrificed and testicles were dissected and processed by routine histological protocols. FSH and LH serum levels were determined by radioimmunoassay. The visualization of antigens was achieved by the streptavidin-peroxidase immunohistochemical method. Antibodies against chromogranin A (CrA), S-100 protein (S-100), P substance (PS), synaptofisin (SYN), neurofilament protein (NF), gliofibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were used. The mean LH and FSH serum concentrations were consistently suppressed with hormonal treatments. Intermediate filaments (NF and GFAP) showed no difference in their expression. The expression of S-100, NSE and SYN was significantly lower in both hormone-treated groups. In oestradiol-treated rats, the immunoreactivity of CrA and SP decreased significantly but was restored after testosterone supplementation. Although the nature and functions of many of these substances in Leydig cells remain unknown, these results are consistent with the hypothesis that the expression of some neuroendocrine markers is hormonally controlled.     

Tissue Culture of the Enteric Nervous System from Equine Ileum

Ileal samples were harvested fresh from euthanized adult horses. The tissues were microdissected to prepare wholemount preparations for immunohistochemistry and for either explant or dissociated culture systems of the enteric nervous system. Explant culture systems were established using wholemounts of either the submucous plexus or the muscularis externa (including the myenteric plexus). Dissociated cell cultures could only be obtained from the submucous plexus. Culture systems were maintained for up to 5 days. Immunoreactivity for a neuronal marker (Pan-N) and for glial cell markers (GFAP and S100) indicated the presence of both neurons and enteric glia in the tissue culture preparations.This is the first report of equine enteric neurons being grown in tissue culture. Further refinements to the techniques will be required before this in vitro model can be used for quantitative analysis.     

Tattooing of the Equine Eyelid: A Retrospective Study

In this retrospective study, medical records of 26 horses whose eyelids were tattooed were analyzed, and data regarding age, breed, eyelid pigment, eye color, sex, and reproductive status of the horses were compiled. Owners of the horses were contacted by telephone and questioned about their horses. Twenty-six horses were tattooed at Colorado State University (CSU), and one of them was tattooed twice. Records were available for 26 horses, and follow-up results by telephone were completed for 22 of those. Telephone follow-up times ranged from 3 months to 17 years after tattooing. American Paint Horses constituted 62% (16/26) of the cases, followed by Appaloosas (19% = 5/26), and Quarter Horses (8% = 2/26). In 100% (22/22) of cases, the reason for presenting the horse for a tattoo was lack of periocular pigmentation. Only two horses developed ocular or periocular squamous cell carcinoma (SCC) after their tattoos. Although the results of this study suggest that tattooing may be protective against SCC and solar blepharitis, it is by no means conclusive. However it appears to have no deleterious effects, and often owners like the appearance of the tattooed eyelid.