大口黑鲈外周血细胞显微结构、细胞化学特征及吞噬作用的研究

通过对大口黑鲈外周血细胞的显微结构及细胞化学染色特征和吞噬作用的研究,为大口黑鲈免疫学研究积累资料。Wright's染色表明:大口黑鲈外周血细胞分为红细胞、粒细胞、单核细胞、淋巴细胞和血栓细胞,其中粒细胞又分为Ⅰ型、Ⅱ型和Ⅲ型。外周血中成熟红细胞最多,占血细胞总数的98.12%,其次为淋巴细胞和血栓细胞,占白细胞中的比例分别为60.92%和22.99%;单核细胞最大,大小为(9.89±0.70)μm×(8.72±0.68)μm;小淋巴细胞最小,大小为(3.88±0.46)μm×(3.48±0.40)μm。大口黑鲈外周血细胞的免疫相关酶ACP、AKP、PO、POX及细胞成分PAS反应、SBB染色的结果表明,所有红细胞的细胞化学染色均呈阴性,不同白细胞的细胞化学染色特征存在差异。其中所有白细胞的AKP、POX染色均呈阴性,PAS反应均呈阳性,除Ⅱ型粒细胞外所有白细胞的PO、ACP染色均呈阳性,除单核细胞外所有白细胞的SBB染色均呈阳性。大口黑鲈外周血细胞吞噬酵母菌的实验表明,红细胞能够吞噬酵母菌,其吞噬率为(15.70±1.07)%,也观察到部分白细胞吞噬酵母的现象。PO和ACP及脂类和糖类可能是大口黑鲈外周血细胞吞噬作用的重要酶类和能量来源。     

养殖齐口裂腹鱼外周血细胞显微观察

养殖齐口裂腹鱼Schizothorax prenanti外周血经瑞氏染色液染色,可鉴定出红细胞、嗜中性粒细胞、血栓细胞、淋巴细胞、单核细胞,一些未成熟的红细胞和少量进行无丝分裂的红细胞,未观察到嗜碱性粒细胞和嗜酸性粒细胞.在外周血中还可观察到3个阶段的嗜中性粒细胞:中性晚幼粒细胞胞体为圆形或近圆形,胞浆呈淡粉红色,量丰富,其中含有较多红色的细小颗粒,胞核呈肾形;中性杆状核粒细胞胞核较中性晚幼粒细胞的凹陷更强,呈"S"、"C"等形状;中性分叶核粒细胞占多数,胞体近圆形,核至少分成两叶,多达五或六叶.血栓细胞呈圆形、近圆形、梨形、纺锤形、杆状等多种形态,一个或多个聚集在一起.外周血细胞中,血栓细胞体积最小、数量最多,单核细胞体积最大、数量最少.     

乌龟外周血细胞的显微和超微结构

乌龟血细胞的显微和超微结构研究表明;在外周血细胞中,可分辨出红细胞、单核细胞、淋巴细胞、血栓细胞、嗜酸性粒细胞、嗜碱性粒细胞、嗜中性粒细胞等7种细胞.红细胞核圆形,胞质均匀无细胞器.单核细胞的特征是核内异染色质多聚集于周边,胞质中含有许多囊泡.淋巴细胞的核质比例大.血栓细胞以具细长的指状突起,基本无细胞器为其特征.嗜酸性粒细胞仅含一种圆形颗粒,颗粒质地均匀,电子致密,大小不等.嗜碱性粒细胞颗粒环绕在胞核周围,有三种电子密度、颗粒大小不一的类型.嗜中性粒细胞含有两种电子密度、形态不一的颗粒.       

花尾胡椒鲷血细胞在光镜和扫描电镜下的结构

本文以花尾胡椒鲷血细胞进行光镜及扫描电镜观察。结果在血涂片中可分辨出红细胞、淋巴细胞、单核细胞、嗜酸性料细胞。嗜中性粒细胞与血栓细胞,没有发现嗜碱性粒细胞。白细胞中,以淋巴细胞的比例最高,嗜酸性粒细胞数量最少。在扫描电镜下,红细胞椭圆形,表面光滑,无突起;而各种白细胞为圆形,细胞表面不平,具有各种突起,揭示白细胞具有活跃的变形运动的吞噬能力。还见到红细胞、血栓细胞可在外周血液中通过直接分裂产生。     

北部湾棱皮树蛙外周血细胞显微结构观察

本文以人们极少关注的两栖动物北部湾棱皮树蛙(Thelodermacorticale)为研究对象,采用瑞氏-姬姆萨混合染色法与血细胞计数法观察并统计了北部湾棱皮树蛙外周血细胞的形态特征和数量比例。结果显示,北部湾棱皮树蛙的外周血由红细胞、嗜中性粒细胞、嗜碱性粒细胞、嗜酸性粒细胞、淋巴细胞(包括大、小淋巴细胞)和单核细胞、血栓细胞7种细胞组成。其中,红细胞的数量最多,平均含量为2.43×10~5个/mm~3,多呈椭圆形或卵圆形,少数呈泪滴形、梨形、逗号形、梭形,具椭圆或圆形核,核染色质密集,偶尔可观察到正在进行直接分裂或有丝分裂的红细胞。白细胞的数量比红细胞少,平均含量为1.74×10~4个/mm~3,其中,小淋巴细胞占40.02%±1.77%、大淋巴细胞占10.83%±2.53%、单核细胞占23.17%±3.16%、嗜中性粒细胞占10.08%±4.62%、嗜碱性粒细胞占9.25%±2.69%、嗜酸性粒细胞占6.72%±1.50%。血栓细胞呈纺锤形和梭形,常单独存在,或者呈圆形和不规则圆形,数个集合在一起。     

海鳗外周血细胞的显微结构

对海鳗外周血液有形成分用常规Wright氏和Giemsa氏染色并进行显微观察,可鉴别出红细胞、血栓细胞、淋巴细胞、单核细胞、中性粒细胞和酸性粒细胞;还见到幼稚的、正在分裂的、分解和解体状态的红细胞;未发现碱性粒细胞。红细胞数量多,椭圆形,具椭圆形核;白细胞中血栓细胞最多,分散分布,形态多样;淋巴细胞、单核细胞、中性和酸性粒细胞与其它鱼类的基本相似。     

史氏鲟外周血细胞的显微及超微结构

采用血细胞计数、光学显微及电子显微技术对二龄史氏鲟外周血细胞的数目、形态及结构进行了研究。二龄史氏鲟红细胞的数目为47.75×104个/mm3,白细胞数目为2.9万个/mm3,其中淋巴细胞所占比率最高。史氏鲟的外周血中除正常红细胞外,还有处于分裂状态及未成熟的红细胞。史氏鲟外周血中的白细胞有四种类型,分别为淋巴细胞、粒细胞、血栓细胞和单核细胞。其中粒细胞有两种,即嗜中性粒细胞和嗜酸性粒细胞。嗜中性粒细胞含有多种形状的核,其中分叶的核数目较多,粒细胞及淋巴细胞均类似于哺乳动物。对史氏鲟外周血细胞细微结构的观察显示:红细胞中具有少量的细胞器;淋巴细胞结构典型;单核细胞较粒细胞稍小且具有较多线粒体;血栓细胞具有梭形和圆形两种,胞质较少,其中梭形的血栓细胞胞质几乎透明;对粒细胞的颗粒按照形状和电子密度进行了分类。     

中华蟾蜍血细胞的观察

应用常规血涂片Wright氏染色、活体观察和血细胞计数法研究中华蟾蜍（Bufo　gargarizans）的血细胞形态。结果表明:红细胞有有核和无核两种类型。有核红细胞较大,椭圆形,平均19.41×14.25μm,细胞核染色质密集。白细胞圆形或椭圆形,直径比红细胞长径短,核比例一般较大。白细胞中,淋巴细胞最多,其次为中性粒细胞和单核细胞,嗜酸性粒细胞少,嗜碱性粒细胞极少。红细胞和淋巴细胞均有直接分裂现象。红细胞平均值为7.67×105/mm3,白细胞平均值为0.75×105/mm3。血栓细胞纺锤形或卵圆形,胞质少或裸核状,推侧为红细胞的衍生物。     

红腹锦鸡血细胞的光镜和扫描电镜观察

为了探讨红腹锦鸡(Chrysolophus pictus)血细胞的形态特征,为生理学研究提供生物学基础资料,利用光镜和扫描电镜观察了红腹锦鸡血细胞的形态特征。结果表明,红腹锦鸡红细胞呈椭圆形或扁圆形,表面光滑,具核;白细胞为球形,体大,淋巴细胞表面有绒毛状突起,嗜中性粒细胞核一般分2~5叶,嗜酸性粒细胞核一般分2叶,嗜碱性粒细胞核分2~3叶,单核细胞表面粗糙不平,核大,呈肾形或圆形;凝血细胞呈球形或不规则形。     

淡水石斑外周血细胞显微结构观察

淡水石斑(Cichlasoma managuense)外周血细胞可区分出红血细胞、嗜中性粒细胞、嗜酸性粒细胞、单核细胞、淋巴细胞和血栓细胞,未发现嗜碱性粒细胞。外周血液中还存在少量未成熟的和正在分裂的红血细胞。白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;数量上,血栓细胞最多,而嗜酸性粒细胞则最少。     

Cytochemical, immunocytochemical and ultrastructural observations on leukocytes and thrombocytes of fat snook (Centropomus parallelus)

The cytochemical, immunocytochemical and ultrastructural characteristics of leukocytes and thrombocytes in the peripheral blood of the fat snook (Centropomus paralellus) - a fish occurring in Brazil - were investigated. The cytochemical methods were performed to demonstrate four enzymatic reactions - o-toluidine-hydrogen peroxide, naphtol AS-MX phosphate, naphtol AS-BI phosphate and alpha-naphtil acetate to detect myeloperoxidase (MPO), alkaline phosphatase (ALP), acid phosphatase (ACP) and non-specific esterase (α-NAE), respectively - and two non-enzymatic ones - Periodic-Acid Schiff (PAS) and Sudan black B (SBB) to detect the occurrence of glycogen and phospholipids, respectively. Immunocytochemical method utilizing polyclonal rabbit antibody against mammal metalloproteinases (MMPs) 2 and 9 were done. Standard method for Electron Microscopy (EM) was applied for the ultrastructural study. The cytochemical reactions were positive in neutrophils for MPO, ACP, α-NAE, glycogen and phospholipids; in lymphocytes for ACP and α-NAE; in monocytes for ACP and α-NAE and in thrombocytes for ACP, α-NAE and glycogen. Only neutrophils were positive for MMPs 2 and 9, and none of the cells studied were positive for ALP. Ultrastructurally: 1) neutrophil showed a spherical shape with a spherical, indented or lobulated euchromatic nucleus, and cytoplasm containing granules of varied sizes and mitochondria of varied shapes and sizes. The nucleus/cytoplasm relation and the size of granules suggest neutrophil maturation in peripheral blood; 2) lymphocytes showed partially heterochromatic nucleus and minimal cytoplasm; 3) monocytes had long cytoplasmic projections, an indented nucleus, evident nucleolus and cytoplasm with granules of varied sizes and vacuoles; 4) thrombocytes were predominantly elliptical or roughly spherical in shape, had a partially heterochromatic nucleus and cytoplasm containing electron-dense granules, intricate canalicular system and vacuoles occasionally holding phagocytic material.     

微观辨证学之研究：气血辨证与细胞化学含量的关系

在临床上为揭示肝脏病肝纤维化气虚血瘀和气滞血瘀病者,对细胞化学含量的变化,运用显微分光光度技术分析,结果发现气虚血瘀者单核细胞ＡＮＡＥ、ＡＣＰ,红细胞ＬＤＨ和嗜中性粒细胞ＡＤＰ明显降低,气滞血瘀者单核细胞ＡＮＡＥ、红细胞ＬＤＨ及嗜中性粒细胞ＡＤＰ较正常组偏低,但却较气虚血瘀组为高,单核细胞ＡＣＰ高于气虚血瘀组和正常对照组．据此,可将气虚、气滞血瘀及正常对照３组区别开来．     

Cytochemical identification of the leukocytes of torpedoes (Torpedo marmorata Risso and Torpedo ocellata Rafinisque)

The authors subjected peripheral blood smears of Torpedoes to cytochemical analysis of lipids, protein, neutral and acid polysaccahrides and of some enzymatic activities, i.e. adenosine triphosphatase (ATP-ase), acid and alkaline phosphatase, aliesterase and peroxidase. It was found that neutrophilic granulocytes are intensely PAS and aliesterase positive and weakly ATP-ase positive. Eosinophilic granulocytes show the presence of neutral polysaccharides in the matrix (which is PAS positive) and strong ATP-ase and acid phosphatase activities in the granules. Lymphocytes sometimes contain weakly PAS and aliesterase positive granules. Monocytes show some small PAS positive granules and weak acid phosphatase and aliesterase activities. Thrombocytes contain some peripheral granules which are PAS positive and slightly ATP-ase positive. There are no transitional forms between the various cellular types. The results confirm the classification of leukocytes of Torpedoes into neutrophilic granulocytes, eosinophilic granulocytes, lymphocytes, monocytes and thrombocytes and contribute some informations about the histoenzymatic content of Elasmobranch leukocytes.     

A morphological and cytochemical study of erythrocytes, thrombocytes and leukocytes in four freshwater teleosts

Erythrocytes, thrombocytes and leukocytes morphology and cytochemical staining were studied in big head carp Aristichthys nobilis , oscar Astronotus ocellatus , traíra Hoplias malabaricus and lambari Astyanax bimaculatus . Reticulocytes contained a granular material similar to residual RNA following staining with brilliant cresyl blue. Neutrophils, lymphocytes and monocytes were morphologically similar in all the four species. Thrombocytes were present in all the four species and were predominantly fusiform, whereas eosinophils occurred only in A. ocellatus . Aristichthys nobilis contained a leukocyte with unstained granules following Romanowsky-type staining, which stained intensely with periodic acid-Schiff (PAS). Glycogen granules were present in thrombocytes, neutrophils and eosinophils but not in monocytes or lymphocytes. Peroxidase staining was observed in neutrophils of A. ocellatus , H. malabaricus and A. bimaculatus but not in A. nobilis . Monocytes of A. ocellatus , H. malabaricus and A. bimaculatus stained positively for non-specific esterase, whereas those of A. nobilis did not stain. Thrombocytes and leukocytes in all four species were negative for alkaline phosphatase. Neutrophils of A. ocellatus and H. malabaricus may be involved in respiratory burst and may play an important microbicidal role.     

Determination of acid α-naphthyl acetate esterase enzyme activity in peripheral blood leukocytes of gazelles (Gazella subgutturosa)

We examined gazelle peripheral blood leucocytes using the α-Naphthyl acetate esterase (ANAE) staining technique (pH 5.8). Our purpose was to determine the percentage of ANAE positive lymphocytes. The proportion of ANAE positive T-lymphocytes was 72%. T-lymphocytes showed an ANAE positive reaction, but eosinophilic granulocytes and monocytes also showed a positive reaction. By contrast, no reaction was detected in B-lymphocytes, neutrophil granulocytes or platelets. The reaction observed in T-lymphocytes was a red-brown coloration, usually 1–2 granules, but enough granules to fill the cytoplasm were detected rarely. As a result of ANAE enzyme staining, we concluded that the staining technique can be used as a cytochemical marker for gazelle T-lymphocytes.     

版纳鱼螈外周血细胞观察

以濒危两栖动物版纳鱼螈(Ichthyophis bannanica)为材料,应用瑞氏-姬姆萨混合染色法与血细胞计数法观察并统计了版纳鱼螈各种外周血细胞的形态特征和数量比例.结果表明,版纳鱼螈的外周血液中红细胞数量较多,呈卵圆形、椭圆形、梭形和梨形,平均含量为2.57 ×105个/mm3.白细胞数量较少,多呈近圆形,平均含量为0.72×103个/mm3.白细胞中,淋巴细胞最多,其次为单核细胞、嗜中性粒细胞、嗜碱性粒细胞和嗜酸性粒细胞.血栓细胞数量较少,常数个集合在一起.同时,将此研究结果与鱼类、爬行类和其他两栖类的血细胞比较,进而探讨了版纳鱼螈的进化地位.     

Investigation of α-naphthyl acetate esterase and acid phosphatase in the peripheral blood leukocytes of greyhounds

The purpose of our study was to determine the percentages of α-naphthyl acetate esterase (ANAE)- and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL), the presence of the ANAE and ACP enzymes in leukocytes, and the proportion of PBL in greyhounds. Peripheral blood samples were collected from the cephalic antebrachial vein of 14 (7 animals of each sex) healthy 1-2-year-old greyhounds. Mean percentages of ANAE-positive PBL were found to be 73.29 ± 0.95% in female and 74.29 ± 2.21% in male dogs. The difference between mean values of the genders was not statistically significant. The ACP values were 36.00 ± 2.94% for females and 33.57 ± 2.15% for males. No significant differences were found with regard to gender. For both enzymes, although monocytes and eosinophilic granulocytes displayed a positive reaction, neutrophils gave negative reactions. The proportion of PBL was 36.29 ± 5.31% and 33.00 ± 2.38 % in female and male dogs, respectively. The differences were not significant.     

Investigation of α-naphthyl acetate esterase and acid phosphatase in the peripheral blood leukocytes of greyhounds

The purpose of our study was to determine the percentages of α-naphthyl acetate esterase (ANAE)- and acid phosphatase (ACP)-positive peripheral blood lymphocytes (PBL), the presence of the ANAE and ACP enzymes in leukocytes, and the proportion of PBL in greyhounds. Peripheral blood samples were collected from the cephalic antebrachial vein of 14 (7 animals of each sex) healthy 1-2-year-old greyhounds. Mean percentages of ANAE-positive PBL were found to be 73.29 ± 0.95% in female and 74.29 ± 2.21% in male dogs. The difference between mean values of the genders was not statistically significant. The ACP values were 36.00 ± 2.94% for females and 33.57 ± 2.15% for males. No significant differences were found with regard to gender. For both enzymes, although monocytes and eosinophilic granulocytes displayed a positive reaction, neutrophils gave negative reactions. The proportion of PBL was 36.29 ± 5.31% and 33.00 ± 2.38 % in female and male dogs, respectively. The differences were not significant.     

Cytochemistry of normal and leukaemic lymphocytes: a review.

Findings with six cytochemical reactions demonstrable in normal and leukaemic lymphocytes were reviewed. The two methods which are presently of greater diagnostic value are the acid phosphatase (AP) and alpha-naphthyl acetate esterase (ANAE) reactions. AP has a definitive role in the diagnosis of acute and chronic T-cell leukaemias, where a strong positive reaction helps to distinguish them from most B-cell lymphoproliferative disorders. New findings concerning the ultrastructural localization of this enzyme are presented. ANAE is of value in distinguishing T-lymphocytes (positive localized reaction) from B lymphocytes (negative reaction) and the T micron from the T gamma subpopulation of T-lymphocytes, a positive reaction demonstrable only in the T micron cells. Other reactions reviewed were PAS, beta-glucoronidase, hexosaminidase and alkaline phosphatase.     

人参不同部位皂甙对衰老的人胚肺成纤维细胞影响的细胞化学研究

应用细胞化学的定性、定位,显微分光光度计定量分析的方法,测定人参不同部位皂甙对低代龄和高代龄人胚肺成纤维细胞内多糖类(PAS反应)、酸性非特异性酯酶(ANAE)、碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、单胺氧化酶(MAO)含量的影响,实验结果表明:人参根、果、茎叶皂甙(SRG、SFG、SSLG)使低代龄细胞的多糖类、MAO含量下降或上升,对ANAE台量没有显著影响。但对高代龄细胞,SFG、SRG、SSLG则显著增加了多糖类、ANAE、ALP、ACP的相对含量,同时降低了MAO的含量。本文提示,人参不同部位皂甙均可以提高衰老细胞内多糖类,ALP、ACP、ANAE的相对含量,降低MAO的含量,而对年青细胞的影响则很不一致。