禁食对鹅腺胃中Ghrelin mRNA表达的影响

为了研究禁食是否对吉林白鹅腺胃Ghrelin mRNA表达有影响,试验采用相对荧光定量方法,以β-actin为内参,检测70日龄吉林白鹅腺胃组织中的Ghrelin mRNA表达量。结果表明:禁食48 h后鹅腺胃Ghrelin mRNA表达水平显著高于自由采食组(P0.05)。说明禁食是影响鹅腺胃Ghrelin mRNA相对表达量的重要因素。     

Ghrelin阳性细胞在食蟹猴消化系统中的分布定位

目的:研究ghrelin阳性细胞在食蟹猴消化系统中的分布定位。方法:采用免疫组化SABC法检测ghrelin免疫阳性细胞在食蟹猴消化系统中的分布部位、含量及细胞形态。结果:ghrelin免疫阳性细胞分布于食蟹猴的胃、十二指肠、空肠前段、空肠中段以及胰腺中,而在口腔黏膜、食管、空肠后端至直肠、肝脏中均无表达。结论:食蟹猴内ghrelin阳性细胞的分布与人和其他动物的分布基本相似,ghrelin在进化上具有保守性,物种间存在差异性。     

GHSR-1a在奶山羊胃肠道的分布与表达

试验采用免疫组织化学、Real-time PCR和Western blotting方法测定ghrelin的功能性受体GHSR-1a(Growth hormone secretagogue receptor-1a,GHSR-1a)在奶山羊胃肠道的分布和表达。免疫组织化学结果显示,GHSR-1a免疫阳性细胞广泛分布于奶山羊胃肠道。在皱胃主要定位于黏膜层和肌层;瘤胃、网胃和瓣胃黏膜层及肌层中也可见GHSR-1a免疫阳性细胞;在小肠主要位于十二指肠、空肠和回肠的黏膜层、黏膜下层和肌层;在结肠、盲肠和直肠GHSR-1a免疫阳性细胞也有广泛分布;GHSR-1a主要表达于内在神经丛神经细胞、胃底腺上皮细胞、肠腺上皮细胞、复层鳞状上皮细胞、平滑肌细胞中。real-time PCR和Western blotting结果显示,皱胃、十二指肠、空肠、回肠、盲肠、结肠和直肠GHSR-1a的表达水平相对较高,显著高于瘤胃、网胃和瓣胃的表达(P<0.05)。结果表明,ghrelin可能通过GHSR-1a对奶山羊胃肠功能具有重要的调节作用。     

雄性食蟹猴生殖系统中Ghrelin的分布定位

本文采用免疫组织化学染色方法,探讨ghrelin在雄性食蟹猴生殖系统内的分布定位。通过对ghrelin免疫阳性细胞在生殖系统中分布部位、含量及细胞形态等方面的研究,为今后ghrelin在食蟹猴体内的功能研究奠定形态学基础。免疫组化染色发现,ghrelin阳性细胞在食蟹猴的生殖系统中有分布。Ghrelin免疫阳性细胞被染为棕色到棕黑色,主要分布于睾丸、附睾及输精管中,精囊腺中无ghrelin阳性细胞分布。Ghrelin阳性细胞在组织中多呈散在分布。细胞大小不一、形态各异,多呈圆形、卵圆形、锥体形、长柱形及其他不规则形。     

GHSR—1a在奶山羊胃肠道的分布与表达

试验采用免疫组织化学、Real—timePCR和Western blotting方法测定ghrelin的功能性受体GHSR-1a（Growth hormone seeretagogue receptor-1a,GHSR-1a）在奶山羊胃肠道的分布和表达。免疫组织化学结果显示,GHSR—1a免疫阳性细胞广泛分布于奶山羊胃肠道。在皱胃主要定位于黏膜层和肌层;瘤胃、网胃和瓣胃黏膜层及肌层中也可见GHSR-1a免疫阳性细胞;在小肠主要位于十二指肠、空肠和回肠的黏膜层、黏膜下层和肌层;在结肠、盲肠和直肠GHSR—1a免疫阳性细胞也有广泛分布;GHSR—1a主要表达于内在神经丛神经细胞、胃底腺上皮细胞、肠腺上皮细胞、复层鳞状上皮细胞、平滑肌细胞中。real—timePCR和Westernblotting结果显示,皱胃、十二指肠、空肠、回肠、盲肠、结肠和直肠GHSR—1a的表达水平相对较高,显著高于瘤胃、网胃和瓣胃的表达（P〈0．05）。结果表明,ghrelin可能通过GHsR-1a对奶山羊胃肠功能具有重要的调节作用。     

蓝狐消化道5-羟色胺和生长抑素免疫组化定位

采用免疫组织化学SP法.研究蓝狐消化道内5-羟色胺和生长抑素2种免疫阳性细胞的形态结构与分布密度.结果表明,消化道中这2种免疫阳性细胞形态多呈圆形、椭圆形或锥体形,主要集中分布在胃腺上皮、肠上皮及肠腺上皮细胞之间.5-HT免疫阳性细胞数量以结肠最多,直肠和空肠次之,胃底腺区、十二指肠、回肠和幽门腺区较少,食管、贲门腺区和盲肠中未见:SS免疫阳性细胞大量出现于幽门腺区,食管、责门腺区和盲肠中未见;根据其形态推测,蓝狐消化道这2种免疫阳性细胞有内、外2种分泌功能.     

禁食对开产蛋鸡红细胞免疫和外周血淋巴细胞免疫功能的影响

为了研究禁食对家禽免疫功能的影响,试验选择90只海兰褐开产蛋鸡为研究对象,随机分为3组,其中一组为常规饲养对照组,试验1组为断料自由饮水(水中含3%葡萄糖+复合多维),试验2组为断料自由饮水并添加免疫增强剂(水中含3%葡萄糖+复合多维+淋巴因子),时间为7 d。分别于禁食前1天、禁食后第4天、禁食后第7天测定试验鸡常规血象、外周血淋巴细胞转化能力、红细胞免疫功能。结果表明:至禁食第4天,与对照组相比,试验1组和2组鸡红细胞和白细胞数量均明显增多,外周血淋巴细胞增殖转化能力和红细胞免疫功能也有提高。说明短期合理禁食可提高开产蛋鸡的免疫功能。     

成年皖西白鹅生殖系统ghrelin的免疫组化定位

为了研究皖西白鹅生殖系统内是否有产生Ghrelin细胞分布，应用免疫组化SABC法并结合DAB显色技术，结果：①在卵巢的生长卵泡中，均有ghrelin免疫阳性细胞，尤其在生长卵泡细胞的颗粒层更为明显；闭锁卵泡的卵泡外腺细胞也呈现ghrelin免疫阳性反应；②在输卵管五段的粘膜层中均观察到ghrelin免疫阳性细胞，其中，漏斗部细胞着色最深，数量最多，膨大部细胞反应最微弱，峡部、子宫部和阴道部ghrelin免疫反应介于二者之间，肌层和外膜均未见ghrelin表达。结论：产生Ghrelin的细胞在成年皖西白鹅卵巢和输卵管中均有广泛的分布，揭示Ghrelin可能调节生殖功能。     

猪胃生长抑素阳性细胞和盐酸细胞发育的研究

应用免疫组织化学技术和组织学方法分别研究猪胃生长发育过程中生长抑素(SS)阳性细胞和盐酸细胞数量的变化。结果表明,胃底腺盐酸细胞数量刚出生时很少,但从3日龄开始一直处于上升趋势,直到120日龄才开始下降。从出生1～3日龄,盐酸细胞数量呈极显著增加(P<0 01);在20日、45日龄和90日龄时,盐酸细胞数量仍均呈显著或极显著增加(P<0 05,P<0 01)。然后于120日龄,盐酸细胞数量呈极显著减少(P<0 01)。胃底腺中SS阳性细胞发育与盐酸细胞正相反,从刚出生(1日龄)到3日龄,SS阳性细胞极显著增加(P<0 01),然后从3日龄以后SS阳性细胞逐渐减少,一直持续到120日龄。表明猪胃底腺中SS阳性细胞的发育可能对盐酸细胞的发育具有调节作用。     

年龄差异对家兔CD4+T细胞在胃肠分布的影响

为深入了解家兔消化道中CD4⁺T细胞的组织学定位以及在消化道不同解剖学部位的数量变化,本试验运用免疫组织化学方法,探讨了家兔胃(胃底腺区、幽门腺区)和小肠及大肠不同解剖学部位中的CD4⁺T细胞定位特点和数量变化规律。结果表明,在胃肠中,CD4⁺T细胞主要分布于固有层。胃底腺区阳性细胞数目要明显比幽门腺区少。小肠、大肠各段的阳性细胞分布数量未见明显差异,由十二指肠到回肠表现为小幅度递增,由盲肠到直肠表现为小幅度递减。胃肠中CD4⁺T细胞在各部位的数量随年龄增长表现出先增后减的变化趋势,老年组和幼年组CD4⁺T细胞数量在大肠中未见明显差异。结果提示,CD4⁺T细胞在青年家兔胃肠中的分布最为丰富。     

Predominant expression of growth hormone secretagogue receptor in the caudal lobe of chicken pituitary and parallel developmental increase in expression of pituitary GH and proventriculus ghrelin mRNA

To examine the involvement of ghrelin in growth hormone (GH) synthesis in the chicken pituitary, the regional distribution of GH secretagogue receptor (GHS‐R)/ghrelin receptor was investigated. Quantitative real‐time polymerase chain reaction (Q‐PCR) analysis revealed that the expression levels of GHS‐R and GH mRNA in the caudal lobe were about fourfold and sevenfold higher in the cephalic lobe of 7 day‐old chickens, respectively. Immunohistochemical analysis showed that GHS‐R immunoreactivity was more abundant in the caudal lobe than in the cephalic lobe, as was the case for GH immunoreactivity. By Q‐PCR, parallel increases were observed in the expression levels of ghrelin mRNA in the proventriculus and GH mRNA in the pituitary from embryonic day 17 to day 7 after hatching, whereas no significant change was found in the expression levels of GHS‐R mRNA in the pituitary during this period. These results suggest that proventriculus‐derived ghrelin may participate in pituitary GH synthesis by acting on its receptor during late embryonic development and the early post‐hatching period in chickens.     

Changes in ghrelin levels of plasma and proventriculus and ghrelin mRNA of proventriculus in fasted and refed layer chicks

This is a test-report of ghrelin levels in plasma and proventriculus, the glandular portion of the avian stomach, by using a specific radioimmunoassay for acylated ghrelin, as well as the expression of the ghrelin gene in the proventriculus after a 12-h fasting period followed by a 6-h feeding period with 6-day-old layer chicks. After fasting, the plasma ghrelin levels increased from 21.3 ± 4.5 to 32.9 ± 5.0 fmol/ml, but once refed it returned to the control value. After fasting, the ghrelin mRNA and the peptide levels in the proventriculus increased, and ghrelin mRNA levels remained high but once refed the ghrelin content returned to the control level. Furthermore, in order to examine the effect of increased circulating ghrelin on food intake, a bolus intravenous injection of 500 pmol of chicken ghrelin was given to 8-day-old chicks. The ghrelin injection did not cause any significant changes in food intake. These results indicate that the levels of ghrelin and its mRNA with layer chicks are altered according to the feeding state and this in a similar manner as has been observed in mammals. Unlike in mammals, an increase in circulating ghrelin does not cause the promotion of food intake in chicks.     

EFFECT OF ANESTHESIA,POSITIONING, TIME,AND FEEDING ON THE PROVENTRICULUS: KEEL RATIO OF CLINICALLY HEALTHY PARROTS

Healthy, adult Hispaniolan Amazon parrots (Amazona ventralis) were imaged on three occasions to determine the effects of anesthesia, patient rotation, feeding, and short/long‐term temporal factors on the proventriculus:keel ratio. Increasing rotation up to 15° from right lateral resulted in increased inability to measure the proventriculus in up to 44% of birds, meaning that the proventriculus:keel ratio could not be calculated from those radiographs. There was a significant difference between the proventriculus:keel ratio for individual parrots when quantified 3 weeks apart. Despite this difference, all ratios remained within normal limits. No significant effect was identified due to anesthesia, feeding, fasting, or repeated imaging through an 8‐h period. Interobserver agreement for measurability and correlation for the proventriculus:keel ratio values was high. It is recommended that the proventriculus:keel ratio be calculated from anesthetized parrots to attain images in true lateral recumbency. Ratio fluctuations within the normal range between radiographs obtained on different dates may be observed in normal parrots.     

Microstructure Features of Proventriculus and Ultrastructure of the Gastric Gland Cells in Chinese Taihe Black‐bone Silky Fowl (Gallus gallus domesticus Brisson)

To investigate microstructure of proventriculus and ultrastructure of the gastric gland cells from Chinese Taihe black‐bone silky fowl (BSF), the proventriculus of 4‐week‐old BSF was sampled. Conventional histological and transmission electron microscope (TEM) methods were used in this study. The wall of the Taihe BSF proventriculus was consisted of four layers, the mucous, submucosa, muscularis externa and the serosa as others birds. The muscularis externa of the birds' proventriculus contained three layers. Much of the melanin was present in loose connective tissue of lamina propria, submucosa, and muscularis externa unlike others. In addition, the ultrastructure of the gastric gland cells was observed by TEM. There was only one kind of gland cell, for example oxynticopeptic cell in proventriculus of Taihe BSF. The oxynticopeptic cells contained numerous mitochondria, cisternae of rough endoplasmic reticulum (CRER), intracellular canaliculi (IC) that secrete hydrochloric acid and small amounts of pepsinogen granules. The rough endoplasmic reticulum (RER) was irregular cisternae with ribosomes and surrounded tightly the mitochondria along their configuration. The electron‐dense pepsinogen granules were round with various sizes. The neighbouring oxynticopeptic cells were closed up with tight junction and gap junction. The inter‐space between the neighbouring oxynticopeptic cells was stenosis or was filled with electron‐dense extracellular substance. In conclusion, the gastric gland cells of Chinese Taihe BSF proventriculus were only oxynticopeptic cells that secrete hydrochloric acid and pepsinogen, but no parietal cells and chief cells of mammal. The gastric gland cells of proventriculus were underdeveloped compared with those of mammals.     

Effects of fasting and refeeding on plasma concentrations of leptin, ghrelin, insulin, growth hormone and metabolites in swine

The effects of nutritional status, such as fasting and refeeding, on leptin and ghrelin secretion in swine were examined. The swine (n = 4) were fasted for 54 h and plasma hormone levels were measured before, during and after fasting. Plasma leptin and insulin concentrations began to decrease 12 and 6 h into the fasting period, respectively (P < 0.05), and maintained a low level for the remaining period of fasting. Plasma leptin and insulin returned to the pre‐fasting value 6 and 12 h after refeeding, respectively. Plasma ghrelin concentrations showed a nocturnal periodicity during the fasting period; it increased nocturnally at 36 and 42 h into the fasting period (P < 0.05). Plasma growth hormone levels did not show any remarkable changes during the fasting. Plasma glucose levels showed a modest fall during fasting and significantly decreased (P < 0.05) at 24 h into the fasting period, returning to pre‐fasting levels after refeeding. Plasma nonesterified fatty acid levels increased (P < 0.05) at 12 h into the fasting period and returned to the pre‐fasting level 6 h after refeeding. These results indicate that plasma leptin, insulin and ghrelin play an important role in maintaining energy homeostasis in swine. The plasma ghrelin did not continuously increase, but showed nocturnal periodicity during fasting. This may suggest that ghrelin is also involved in physiological processes other than energy homeostasis.     

Proliferative response by the ostrich proventriculus in idiopathic gastric stasis: a case report

In this report, the late clinical, necropsy, and histopathological findings of idiopathic gastric stasis in an ostrich are described, with special attention to the proventriculus. An ostrich with clinical signs of gastric stasis was brought to our clinic in a state of impending death that could not be reversed with emergency medical care. At necropsy, the koilin layer of the gizzard was uneven without ulcerations. The proventriculus was distended with food items without impaction and mucosal lesions. None of the most typical aetiological factors including foreign objects, Macrorhabdus ornithogaster and parasitic infection with Libyostrongylus douglassii was present. Histologically, the proventriculus was characterised by hyperplastic mucosa with luminal buds and necrotic cells in the lumen. Immunochemistry for proliferating cell nuclear antigen (PCNA) revealed a high rate of proliferation in epithelial cells of the glandular and mucosal lining in the proventriculus neighbouring the affected gizzard: approximately 88.6% of the cells were immunoreactive as compared to 34.4% of the cells in control ostriches used for comparison. In conclusion, in the absence of gizzard contraction, the mucosal lining of the proventriculus is hyperplastic with a high rate of proliferation that may help compensate the distension due to the accumulation of food items.     

Distribution and ontogeny of gastrin- and serotonin-immunoreactive cells in the proventriculus of developing chick, Gallus gallus domestica

The ontogeny and distribution of gastrin- and serotonin-immunoreactive (IR) cell in the proventriculus of chicks (Gallus gallus domestica, n = 60) in different growth periods was examined immunohistochemically using antisera specific to gastrin and serotonin. Gastrin and serotonin-IR cells were detected in chick proventriculus. Gastrin-IR cells were first evident after 12 days of incubation in lamina epithelialis and compound glands, while serotonin-IR cells were observed only in compound glands at that same time. Gastrin-IR and serotonin-IR cells increased in frequency on incubation day 14 and 16, respectively. Towards the end of incubation, gastrin- and serotonin-IR cell numbers decreased. In adult chicken, both IR cells were present but not lower numbers. The observations demonstrate the presence of gastrin- and serotonin-IR cells in the proventriculus of developing chicks in temporally changing frequencies.     

The Distribution of Gastrin,Somatostatin and Glucagon Immunoreactive (IR) Cells in Ostrich Stomach During the Pre‐ and Post‐hatching Period

Peptides of the gastrointestinal tract play a significant role in the digestive processes and the development of the body; therefore, it is important to have an understanding of location and distribution of gastrin, somatostatin and glucagon immunoreactive (IR) cells in the stomach mucosa of growing birds. For this purpose, 6 embryos and 37 chicks from an ostrich farm in Latvia were used. Tissue samples were collected from the proventriculus – superficial and deep glandular region and from the ventriculus – side wall and pyloric region. The number of cells was determined in 10 mucosal fields of each tissue sample. For statistical analysis, the one‐way anova method was used. Gastrin IR cells regarding the stomach mucosa were found only in the pyloric region. Somatostatin IR cells were most densely located in the pyloric region too, but some cells were also discovered in the mucosa of proventriculus and ventriculus. Glucagon IR cells were found in the epithelium of the deep glands of the proventriculus and only some cells of the superficial glands of the proventriculus, and the ventriculus side wall mucosa. Gastrin and somatostatin IR cells were present in a comparatively large quantity in the ostrich chicks' ventriculus – pyloric region yet not long before hatching. They were located deep in the mucosa of pyloric glands, and their number tended to increase with birds advancing in age.     

禁食、急性冷暴露和外源性注射Ghrelin对羔羊生理指标的影响

为了探究禁食、急性冷暴露和注射外源性生长激素释放肽Ghrelin对羔羊生理指标的影响,本研究监测了室温禁食24h后继续禁食并冷冻(-24.7℃±0.2℃)2h羔羊基本生命体征、血液生理指标和血糖浓度的变化,同时探究了外源性Ghrelin对于冷冻的禁食羔羊以上指标的影响。结果显示,在禁食后8、16、24h羔羊呼吸频率、心率和血糖浓度均较禁食前显著降低(P0.05),在禁食后16h和24h直肠温度均较禁食前极显著降低(P0.01),而血液生理指标变化不明显(P0.05);在冷冻1.5h和2.0h后羔羊呼吸频率较保温组显著减慢(P0.05),心率和血糖浓度在冷冻后1.0、1.5、2.0h显著增加(P0.05),冷冻过程中羔羊直肠温度差异不显著(P0.05);冷冻2h后颈静脉一次性注射Ghrelin未发现对羔羊以上指标的影响(P0.05)。以上研究结果提示,羔羊在禁食和低温情况下可通过调整自身活动,以维持机体稳态,并且Ghrelin可能并未参与该调整过程。