日粮中缺硒对鸡空肠肥大细胞组织化学和超微结构的影响

为探讨硒缺乏对鸡空肠中肥大细胞(MC)的形态学和超微结构的影响,将10只鸡随机分成低硒试验组(饲喂低硒日粮0.032 mg/kg)和正常对照组,在75日龄时断头处死立即取出空肠,进行甲苯胺蓝、长时间甲苯胺蓝、阿利新蓝-藏花红染色、阿利新蓝-高碘雪夫氏反应和醛品红-阿利新蓝5种染色并进行透射电镜观察。结果显示,采用不同染色方法所获得的MC数量、大小、形态、颜色及颗粒的黏多糖成分均不相同。电镜下,MC细胞胞浆内充盈大量的特征性颗粒,颗粒内容物呈均质状,电子密度高低不等,核圆形或椭圆形,胞质中可见高尔基复合体和较多的线粒体;试验组可见MC数量显著增加并与变性细胞紧密相邻,异染色质增多,核偏位或固缩,胞质稀疏,同时可见MC内颗粒内容物释放后的空隙或空泡。因此,组化染色提示肥大细胞存在不同亚型;试验组MC数量明显增加,这提示MC是重要的免疫细胞之一,在机体的免疫调节中发挥着重要的作用。     

宁都黄鸡小肠显微结构和甲苯胺蓝染色阳性细胞分布特点

通过石蜡切片和HE染色方法观察了宁都黄鸡小肠的显微结构,同时通过改良甲苯胺蓝染色的方法鉴定小肠中的甲苯胺蓝染色阳性细胞.结果显示,宁都黄鸡小肠与一般禽类小肠显微结构类似,黏膜下层缺乏十二指肠腺,黏膜固有层缺乏淋巴小结.小肠从前到后,杯状细胞逐渐增多.3段小肠的肠腺和回肠杯状细胞呈甲苯胺蓝染色阳性.小肠壁四层结构中都有肥大细胞的分布,主要分布于黏膜上皮细胞下、黏膜下层、肌层和外膜结缔组织中.肥大细胞体积较大,多数呈卵圆形,有的细胞形态不规则,甚至有突起.胞核染色呈深蓝色,胞质中含有紫红色颗粒.结果表明,宁都黄鸡小肠腺细胞和杯状细胞甲苯胺蓝染色呈阳性,说明其能够分泌有助于吸收的活性物质;小肠中具有丰富的肥大细胞,说明宁都黄鸡有较完善的肠道免疫屏障.     

不同固定液和染色方法显示绒山羊空肠内肥大细胞效果的比较

将绒山羊的空肠用Carnoy氏液、Bouin氏液、4%的中性甲醛(NBF)及4%的多聚甲醛分别固定,然后进行常规甲苯胺蓝-藏红O复染法(MTB-S)和改良甲苯胺蓝-藏红O复染法(LTB-S)染色,光镜观察,探讨不同的固定液和染色方法对显示绒山羊空肠肥大细胞(MC)的影响。结果:绒山羊空肠组织采用Carnoy氏液固定,甲苯胺蓝-藏红O复染法和改良甲苯胺蓝-藏红O复染法对所有的肥大细胞均有较好的染色效果,而Bouin氏液、4%的中性甲醛(NBF)及4%的多聚甲醛固定的空肠,只有少量的MC显示。说明MTB-S与LTB-S均是绒山羊肥大细胞较好的染色方法,固定液配Carnoy氏液为佳。     

家兔主要黏膜免疫器官肥大细胞的免疫组化及超微结构特征

为了探讨家兔黏膜免疫器官肥大细胞（MC）的分布、超微结构及巴氏杆菌感染后特征，对家兔空肠及圆小囊MC进行不同时间甲苯胺蓝染色与血管活性肠肽（VIP）和／或P物质（SP）免疫组化检查，并进行光、电镜观察。结果发现，采用不同固定液和染色方法所获得的MC数量不同，家兔黏膜免疫器官肥大细胞主要分布于黏膜固有层或黏膜下层结缔组织中；当巴氏杆菌感染时MC数量显著增加，这可能对机体的抗感染有重要意义。电镜下，MC胞浆内充盈大量呈电子密度不同的均质状物基质颗粒，未见特殊超微结构。     

硒缺乏鸡免疫器官中肥大细胞超微结构观察

试验对饲喂低硒日粮(0.032 mg/kg)的鸡脾脏、胸腺、法氏囊进行透射电镜观察,以探讨硒缺乏对鸡免疫器官中肥大细胞(MC)结构变化与功能的影响.结果表明:缺硒组鸡免疫器官中肥大细胞数量显著增加并与变性细胞紧密相邻,肥大细胞胞浆内充盈大量呈现电子密度不同的均质状基质颗粒,同时可见肥大细胞颗粒内容物释放后的空隙或空泡.结果说明硒缺乏可引起肥大细胞脱颗粒并释放生物活性物质,对变性细胞周围组织的免疫机能可能起重要作用.     

家兔黏膜免疫器官肥大细胞在巴氏杆菌感染中的特征——组织化学与电镜观察

对5例巴氏杆菌感染家兔的空肠与圆小囊肥大细胞（mast cell，MC）进行了组织化学与电镜的观察，发现在细菌感染导致的病变组织边缘、病变组织内血管附近结缔组织中及变性细胞周围组织的上皮下与黏膜下层都出现了大量甲苯胺蓝（Toluidine Blue，TB）阳性肥大细胞。电镜下，MC有的直接与变性细胞密切接触，有的则和血管内皮紧密相触，在此部位的血管内可见有淋巴细胞贴附填充。MC胞浆内充盈大量的特征性颗粒，有时这些颗粒向细胞表面突出形成边缘空隙，即形成所谓的脱颗粒管道，将颗粒内容物逐渐排除细胞外，遗留空腔。     

实验性乳腺炎小鼠乳腺肥大细胞的变化

应用改良甲苯胺蓝染色法和苏木精-伊红染色法对金黄色葡萄球菌诱发实验性乳腺炎小鼠乳腺肥大细胞的数量、分布、活性进行了研究。正常对照组小鼠乳腺组织中肥大细胞主要分布于乳腺小叶之间的结缔组织中,有些围绕血管或乳腺导管分布,腺上皮之间没有肥大细胞分布;阳性试验组小鼠乳腺肥大细胞主要分布在相邻腺泡周围或腺导管上皮周围,胞质内充满异染颗粒,脱颗粒状态显著,甚至表现有颗粒耗竭现象,腺泡中有一定程度的肥大细胞浸润。肥大细胞计数结果显示,阳性试验组母鼠乳腺内的肥大细胞数急剧增多,显著高于正常对照组(P<0.05),而脱颗粒肥大细胞数阳性试验组与正常对照组相比极显著增多(P<0.01)。结论:肥大细胞在小鼠乳腺炎的致病机制中发挥重要作用,参与了乳腺炎的发病过程。     

蒙古马皮肤肥大细胞两种染色方法的比较

为探讨不同染色方法对蒙古马皮肤肥大细胞(MC)显示的影响,本试验采用将蒙古马皮肤用Carnoy氏液固定,然后进行常规甲苯胺蓝-藏红O复染法和改良甲苯胺蓝-藏红O不同复染时间的染色方法,光镜下进行观察.结果表明,蒙古马皮肤采用改良甲苯胺蓝-藏红O复染5 s染色方法(MTB-S)对肥大细胞有很好的染色效果.     

两种固定液对甲苯胺蓝染色效果的影响

甲苯胺蓝染色法是目前观察肥大细胞最常用的一种细胞化学染色方法.由于该方法简单便捷,还被广泛运用于卡式肺孢子虫囊壁上的圆括号状小体、血涂片及骨髓涂片中嗜碱性粒细胞、宫颈癌、幽门螺杆菌等的检测 [1-6].通过甲苯胺蓝染色可以在显微镜下清晰分辨出肥大细胞,呈紫红色颗粒,背景呈淡蓝色.甲苯胺蓝染色的方法不断被研究人员改良,以求得到更好的染色效果,但有关不同固定液对甲苯胺蓝染色效果影响的研究少有报道.     

牛蛙皮肤显微结构与组织化学研究

运用HE染色和3种组织化学方法—改良甲苯胺蓝、阿利新蓝-PAS和多巴氧化酶法染色对牛蛙皮肤显微结构和特性进行了研究。结果表明,牛蛙皮肤由表皮和真皮组成,表皮由角质层、颗粒层和生发层组成,表皮内无黑色素分布,微血管较少。真皮比表皮厚,分为疏松层和致密层。真皮疏松层内有大量团块状、致密的黑色素,微血管较多。真皮疏松层中分布着黏液腺和颗粒腺,黏液腺多于颗粒腺。牛蛙皮肤肥大细胞主要分布于真皮疏松层和皮下层,且多为长条形。真皮致密层呈阿利新蓝-PAS染色强阳性,表皮与真皮交界处以及真皮疏松层和致密层交界处也有强阳性带。多巴氧化酶法染色结果表明,无黑色素分布的表皮层有许多弥散、细小的阳性棕黑色颗粒状色素,而有大块天然黑色素分布的真皮疏松层阳性颗粒色素较少。结果提示,牛蛙皮肤可通过腺体、肥大细胞和黑色素发挥重要的免疫防御和保护作用。     

Mast cells of the bovine trachea: staining characteristics, dispersion techniques and response to secretagogues.

Sections of the lower trachea of cattle, fixed in either Carnoy''s or formalin, were stained with toluidine blue, alcian blue, or alcian blue and safranin O to study the mast cell population. After toluidine blue staining, about twice as many cells in tissue fixed in Carnoy''s contained dark blue granules compared with tissue fixed in formalin. In addition, for the first time in cattle, a population of cells containing red granules was identified after staining with alcian blue and safranin O. Most of these red granules were formalin sensitive. An enzymatic dispersal technique for mast cells is described that yielded 9.4+/-0.4% mast cells (percentage of nucleated cells) with a viability of 92.3+/-0.6%. Spontaneous histamine release was 3.3+/-0.8%. Dispersed mast cells were challenged with various immunological and nonimmunological secretagogues. The calcium ionophores, A23187, ionomyocin, and BrX537A, were effective in releasing up to 94% of histamine in mast cells in a dose-response relationship. Pasteurella haemolytica culture supernate caused about 10% histamine release at a dose of 0.5 mg/mL after correction for spontaneous release. The average histamine content of the mast cells was 6.6+/-1.0 pg/cell. Cytospins of dispersed cells fixed in Carnoy''s and stained with alcian blue and safranin O contained mast cells with blue and red granules, and a few cells with a mixture of both granule types. Based on the effects of type of fixation, staining characteristics and histamine content, a mix of subtypes of mast cells is present in the bovine trachea. However, functionally they respond to secretagogues differently than rodent mast cells. Without an immunological secretagogue, studies to determine compounds that will be effective in blocking mast cell degranulation will be limited.     

The distribution and heterogeneity of mast cells in tongue from five different avian species

This study was conducted with the aim of determining the morphology, distribution and heterogeneity of mast cells in the tongues of seagull (Larus fuscus), common buzzard (Buteo buteo), goose (Anser anser), white stork (Ciconia ciconia) and Gerze rooster. The study used five samples of tongue material from each of the healthy adult avian species. The samples were fixed in 10% neutral‐buffered formalin (NBF) solution, then, after routine tissue follow‐up, the samples blocked with paraplast. Cross‐sections with 5–6 μm of thickness were stained with the 0.5% toluidine blue and alcian blue/safranin O (AB/SO). In all five avian species, it was found that the mast cells were in different sizes and round, oval or spindle‐shaped based on their place of distribution. Mast cell numbers were determined in stained with toluidine blue, examined ×40 objectives in a 1 mm² area. It was observed that mast cell density in subepithelial lamina propria and microscopic papilla was higher in the tongues of all species. Mast cell distribution and heterogeneity varied through the tongue, and there were more mast cells in the dorsal side of the tongue than the ventral side. The highest amount of mast cells was found in the tongue of the Gerze rooster among all five species. In the tongue cross‐sections stained with the combined method of alcian blue/safranin O (AB/SO), the mast cells were stained as AB (+), SO (+) and AB/SO (+) (mixed).     

Morphological Investigations of the Glandulae Profundae Plicae Semilunares Conjuctivae in the Domestic Swine (Sus scrofa domesticus) and the Wild Hog (Sus scrofa ferus)

Samples of glandulae profundae plicae semilunares (Harderian glands) from five domestic swine and five wild hogs were used for this research. The gland samples were fixed in Bouin solution and mounted in paraffin. The paraffin slices were stained with haematoxylin and eosin, according to the periodic-acid-Schiff (PAS) method, with alcian blue (pH 2.5), with toluidine blue (pH 4.0), and applying a combination of staining with alcian blue (pH 2.5) and the PAS method. In domestic swine and wild hogs, these glands are tubular-alveolar with wide glandular lumina. A great deal of acid mucopolysaccharides and PAS-positive substances were noted within swines’ glandular cells, while, in wild hogs, PAS-positive substances were not frequent, acid mucopolysaccharides being noted in only a few glandular-acini cells. The appearance of the metachromatic phenomenon was not noted either in domestic swine or in wild hogs. In domestic swine, the level of acid mucopolysaccharides is probably due either to the housing method or to the influence of the alkaline substances that may appear on an eye's mucous conjunctive membrane during intensive breeding, such that the Harderian glands protect the mucous membrane by extracting acid mucopolysaccharides.     

PAS-positive inclusions in the alveolar type II cells of the caprine lung.

Histochemical analysis of inclusions in the alveolar type II cells was done with the alcian blue-PAS technique. PAS-positive inclusions, which were absent in the sheep and buffalo, were detected in the alveolar type II cells of the goat lung. Mixed diamine-sodium chloride did not stain these inclusions which also showed orthochromatic blue reaction with toluidine blue. The high percentage of alveolar type II cells showing these inclusions suggested that this difference could be explained by the environmental habitat of the goat.     

壳聚糖硒对蛋雏鸡生长、组织硒含量及血清肝酶活性的影响

为探索壳聚糖硒对蛋雏鸡生长、组织硒含量及血清肝酶活性的影响,选取135只1日龄SPF蛋雏鸡,随机分为3组:一组为对照组,饲喂基础日粮;二组和三组在基础日粮中分别添加硒水平为0.15 mg/kg的亚硒酸钠和壳聚糖硒。结果显示:14日龄时亚硒酸钠组和壳聚糖硒组的耗料量、血清碱性磷酸酶(AKP)活性和胸肌、心肌、肾脏、全血的硒含量显著高于对照组(P<0.05),壳聚糖硒组心肌和肝脏的硒含量显著高于亚硒酸钠组(P<0.05);28日龄时壳聚糖硒组与亚硒酸钠组的体重、血清AKP活性和肝脏、肾脏、胸肌、心肌、全血的硒含量显著高于对照组(P<0.05),壳聚糖硒组的耗料量和肾脏、心肌、全血的硒含量显著高于亚硒酸钠组(P<0.05)。研究表明:壳聚糖硒能促进鸡的生长和硒的沉积,提高血清AKP活性,且效果优于亚硒酸钠。     

Staining pattern of the brush border and detection of cytoplasmic granules in the uriniferous tubules of female DBA/2Cr mouse kidney: comparison among various fixations and stains.

The proximal straight tubules of the female mouse kidney exhibit heavy periodic acid Schiff (PAS) staining in their brush borders and numerous cytoplasmic granules. In the present study, the female DBA/2Cr mouse kidney was examined, using various fixatives (formalin, PFA, PLP, Zamboni's, Bouin, or Carnoy solution) and various staining methods (HE, PAS, alcian blue, periodic acid methenamine-silver (PAM), toluidine blue, azan, or Congo red). Under azan and PAM, the staining pattern of the brush border was similar to that of PAS, and few effects of the fixative were observed. Cytoplasmic granules were clearly detected with PAM as well as PAS. However, these granules were not detected with Carnoy solution. Furthermore, distribution of granules differed between PAS and PAM.     

Effect of dietary selenium on the development of Fusarium-induced tibial dyschondroplasia in broiler chickens

A trial was conducted to determine the effects of dietary level of selenium on the pathogenesis of Fusarium-induced tibial dyschondroplasia (FITD) in broiler chicks, and to assess the applicability of FITD as an animal model of Kashin-Beck disease of humans. Day-old female broilers were fed diets that were deficient in selenium (0.02 ppm Se), adequate in selenium (0.15 ppm Se), or generous in selenium (0.50 ppm Se). TDP-1, the toxic component of the fungus, was administered to 15 of 26 chicks in each dietary group starting at 1 week of age and continuing until the chicks were killed at 24-30 days of age. Plasma selenium levels and hepatic glutathione peroxidase activity were significantly lower in the selenium-deficient group than in other dietary groups; these parameters were not affected by treatment with TDP-1. The mortality rate of the TDP-1-treated selenium-generous group was significantly less than that in the other TDP-1-treated groups, but there were no differences in the incidence, severity, or character of the FITD lesions among the groups. Thus, the interaction of selenium and TDP-1 did not include an effect on FITD.     

Distribution, density and histochemical profiles of the lung mast cells during the post-hatching period of Japanese quails (Coturnix coturnix japonica)

The distribution, density and histochemical characteristics of mast cells in the lungs of the Japanese quail were investigated during the post-hatching period. In the period starting from the first to the 60th day post-hatching, based on proteoglycan content, three types of mast cells, which were alcian blue-positive, safranin O-positive and alcian blue/safranin O-positive, were found to exist in the lungs. The application of staining with berberine sulphate demonstrated that, similar to the distribution of safranin O-positive cells, the heparin-containing cells were located in the periphery of large blood vessels. The percentages of mast cells in different localization sites of the lungs were found to vary with age in the post-hatching period with toluidine blue staining. The lack of any statistically significant increase/decrease in the number of mast cells per unit area of the right and left lung lobes is partially in favour of the proposal that the mast cell number increases with the growth of the lung volume in the post-hatching period.     

二乙基己烯雌酚对成年仓鼠附睾肥大细胞数量分布的影响

成年雄性仓鼠经皮下连续注射二乙基己烯雌酚(DES)7 d后,用改良甲苯胺蓝染色法(MTB)和阿尔新蓝番红染色法(AB S)研究DES对附睾肥大细胞的形态大小、类型和数量分布的影响。结果表明:仓鼠附睾肥大细胞为结缔组织型肥大细胞,多分布于附睾头与附睾尾的被膜和间质中。试验组的肥大细胞常见于附睾管近旁,多呈脱颗粒状。虽然试验组与对照组的肥大细胞大小相近,但试验组的肥大细胞数量增多,尤其是附睾尾间质中肥大细胞增加明显(P<0.01)。