除草剂丁草胺对泥鳅体内6种代谢与免疫相关酶的抑制作用

目的研究除草剂丁草胺对泥鳅(Misgurnus anguillicaudatus)体内重要器官过氧化物酶(POX)、非特异性酯酶(NSE)、碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、琥珀酸脱氢酶(SDH)和腺苷三磷酸酶(ATPase)等6种代谢与免疫相关酶活力的影响。方法将泥鳅暴露在不同浓度的丁草胺中96 h,解剖取其心脏、肝胰脏、鳃、头肾、胃和肠等6种器官,采用冰冻切片、酶组织化学及光密度分析技术检测酶活力变化。结果POX在头肾中活性显著较高,其余器官中活性均较低;高浓度丁草胺显著抑制头肾POX活性,但显著促进肝胰脏、胃和肠POX活性。NSE在肝胰脏中活性显著较高,心脏中活性显著较低;高浓度丁草胺对除心脏和胃以外的体内器官NSE活性具有显著抑制作用。ALP在胃中活性显著较高,心脏和头肾中未检测出明显的酶活性;高浓度丁草胺显著抑制胃和肠ALP活性,对肝胰脏和鳃ALP活性无显著影响。ACP在胃和肠中活性显著较高,心脏中活性显著较低;高浓度丁草胺显著抑制肝胰脏、鳃、头肾、胃和肠ACP活性,对心脏ACP活性无显著影响。SDH在心脏中活性显著较高,鳃和头肾中未检测出明显的酶活性;高浓度丁草胺显著抑制心脏、肝胰脏和胃SDH活性,对肠SDH活性无显著影响。ATPase在心脏中活性显著较高,头肾和肠中活性显著较低;高浓度丁草胺显著抑制心脏、肝胰脏和头肾ATPase活性,但对胃和肠ATPase活性具有显著促进作用。结论丁草胺可能通过抑制心脏SDH和ATPase活性影响泥鳅的血液循环,通过抑制胃和肠ALP和ACP活性降低泥鳅对营养物质的消化和吸收能力,通过抑制肝胰和头肾NSE、ACP和ATPase活性损害泥鳅的解毒与免疫防御功能。

矩镰荚苜蓿地下器官贮藏组织中淀粉粒变化规律

矩镰荚苜蓿(Medicago archiducis-nicolai)分布于青藏高原东缘高寒地区,具有极强的抗寒能力。为了探明矩镰荚苜蓿地下器官的越冬抗寒机理,本研究采用常规石蜡切片法,对自然越冬过程中根、根颈和地下茎贮藏组织中淀粉粒的变化进行动态观测。结果显示:矩镰荚苜蓿越冬过程中,淀粉主要贮藏在地下器官的髓和栓内层薄壁细胞中,少量存在于木质部薄壁细胞,根颈与根内淀粉粒分布较为密集;地下不同器官中淀粉数量在越冬前后变化趋势基本一致,总体呈现出先减少后增加的变化规律,越冬前期淀粉在地下器官中大量积累,越冬中期逐渐消失,后期又逐渐积累起来;各地下器官中根颈对温度变化响应最快,地下茎对低温最敏感。研究发现,矩镰荚苜蓿地下茎除具有扩展、繁殖能力外,还兼具贮藏与抗寒等能力,是该植物不同于其他苜蓿属物种的特殊地下器官;越冬期矩镰荚苜蓿地下器官抗寒力形成的不同步性,可能是其应对高寒环境的适应性策略。

Myelin histology:a key tool in nervous system research

The myelin sheath is a lipoprotein-rich,multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers.Due to the complex structure and composition of myelin,various histological techniques have been developed over the centuries to evaluate myelin under normal,pathological or experimental conditions.Today,methods to assess myelin integrity or content are key tools in both clinical diagnosis and neuroscience research.In this review,we provide an updated summary of the composition and structure of the myelin sheath and discuss some histological procedures,from tissue fixation and processing techniques to the most used and practical myelin histological staining methods.Considering the lipoprotein nature of myelin,the main features and technical details of the different available methods that can be used to evaluate the lipid or protein components of myelin are described,as well as the precise ultrastructural techniques.

Ultrastructure and histochemistry of rat myocardial capillary endothelial cells in response to diabetes and hypertension

Insufficient growth and rarefaction of capillaries, followed by endothelial dysfunction may represent one of the most critical mechanisms involved in heart damage. In this study we examined histochemical and ultrastructural changes in myocardial capillary endothelium in two models of heart failure streptozotocin-induced diabetes mellitus （STZ） and NO-deficient hypertension in male Wistar rats. Diabetes was induced by a single i.v. dose of STZ （45 mg/kg） and chronic 9-week stage was analysed. To induce NO-deficient hypertension, animals were treated with inhibitor of NO synthase Lnitroarginine methylester （L-NAME） （40 mg/kg） for 4 weeks. Left ventricular tissue was processed for enzyme catalytic histochemistry of capillary alkaline phosphatase （A1Ph）, dipeptidyl peptidase IV （DPP IV）, and endothelial NO synthase/NADPH-diaphorase （NOS） and for ultrastructural analysis. In diabetic and hypertensive rats, lower/absent A1Ph and DPP IV activities were found in focal micro-areas. NOS activity was significantly reduced and persisted only locally. Quantitative evaluation demonstrated reduction of reaction product intensity of A1Ph, DPP and NOS by 49.50%,74.36%, 20.05% in diabetic and 62.93%, 82.71%, 37.65% in hypertensive rats. Subcellular alterations of endothelial cells were found in heart of both groups suggesting injury of capillary function as well as compensatory processes. Endothelial injury was more significant in diabetic animals, in contrast the adaptation was more evident in hypertensive ones. Concluding： both STZ-induced diabetes- and NO-deficient hypertension-related cardiomyopathy were accompanied by similar features of structural remodelling of cardiac capillary network manifested as angiogenesis and angiopathy. The latter was however, predominant and may accelerate disappearance of capillary endothelium contributing to myocardial dysfunction.

High-efficiency somatic embryogenesis and morphohistology and histochemistry of somatic embryo development in Larix leptolepis Gordon

A high-efficiency somatic embryogenesis protocol of Japanese larch （Larix leptolepis Gordon） has been established in our investigation. Calli were induced from immature zygotic embryos of female cones ofL. leptolepis and then subcultured regularly on to a modified Gupta and Durzan （DCR） basal medium for 5 years. Embryogenic tissues showed distinct morphological changes dur- ing somatic embryo development when they were transferred to a maturation medium supplemented with abscisic acid （ABA） com- pared with the morphology in a medium lacking ABA. Histological observations indicated that polyembryony was a characteristic feature during early embryogeny and somatic embryos at later stages showed normal histodifferentiation. In addition, histochemical analysis revealed that abundant starch granules and proteins accumulated in mature embryos, indicating that they played important roles in the development and regeneration of normal plantlets from somatic embryos on hormone-free germination media

Intestinal metaplasia,dysplasia and gastric cancer:a study of mucohistochemistry,immunohistochemistry and cell DNA quantitative analysis

One hundred and sixty-eight specimens of intestinal metaplasia(IM)with variousaccompanying lesions in gastric mucosa were studied with mucohistochemical and ABCimmunohistochemical staining,The quantitative analysis of cell DNA was done withflowcytometry for 36 specimens.The results indicated that the incidence of type Ⅱb IM wassignificantly higher in the groups of dysplasia(34.6％)and mucosa adjacent to gastric cancer(GC)(51.7％)than in the chronic atrophic gastritis(CAG)group(16.0％)(P【0.01).The expres-sion rate of monoclonal antibody MG7 related antigen(MG7-Ag)in type Ⅱb IM(473％)wasalso significantly higher than those in type la(29.7％),Ib(26.1％)and Ⅱa IM(28.3％)(P【0.05).Expression rate of MG7-Ag,DNA aneuploid rate and percentage of S phase cell werestatistically higher in the type Ⅱb IM with dysphsia(62.5％,62.5％ and 143±32)than in typeⅡb 1M without dysplasia(47.3％,12.5％and12.7±2.9)(P【0.05 and P【0.01).These findingssuppor the supposed progressive process:CAG→type Ⅱb IM→dysplasia→GC,andtype Ⅱb IM with dysplasia is closely associated with GC.

Effects of Natural Radiation, PAR and Artificial Ultraviolet Radiation-B on the Ultrastructure and Histochemistry of Leaf of Oryza sativa L.

Ultraviolet radiation-B (UVBR) affects plants in many important ways, including reduced growth rate, reduction of primary productivity and changes in ultrastructure. The rice (Oryza sativa) is one of the most cultivated cereal in the world along with corn and wheat, representing over 50% of agricultural production. In this study, we examined O. sativa exposed to natural radiation denominated which “ambient samples”, plants cultivated which photosynthetically active radiation (PAR), denominated with PAR-only and plants cultivated with PAR + UVBR for 2 h per day during 30 days of cultivation in vitro. The samples were processed for electron microscopy and histochemistry analysis. PAR + UVBR caused changes in the ultrastructure of leaf of O. sativa, mesophyll cells, which included increased thickness of the cell wall and plastoglobuli, reduced intracellular spaces, changes in the cell contour, and destruction of chloroplast and mitochondria internal organization. The exposure to PAR + UVBR led to changes in guard and subsidiary cells, and the stomata and papillae were with irregular shape. The reduction of epicuticular wax that covered the leaf, was observed. Taken together, these ?ndings strongly suggested that PAR + UVBR negatively affects the ultrastructure and morphology and growth rates, of leaf of O. sativa and, in the long term, their economic viability.

A Study of Histology and Enzymatic Histochemistry on Rabbit's Retina in Acute Ocular Hypertension

The changes of activities of enzymes relating to energy metabolism in rabbit's retina in acute ocular hypertension were observed. The activities of succinate dehydrogenase and adenosine triphosphatase were found to be reduced, while the activities of the lactatic dehydrognease and glucose-6-phosphatase increased. The results revealed the metabolic disturbance of energy in retina after acute ocular hypertension might be the underlying factors relating to the defects of the functions and structures of the...

Histochemistry of microinfarcts in the mouse brain after injection of fluorescent microspheres into the common carotid artery

The mouse model of multiple cerebral infarctions,established by injecting fluorescent microspheres into the common carotid artery,is a recent development in animal models of cerebral ischemia.To investigate its effectiveness,mouse models of cerebral infarction were created by injecting fluorescent microspheres,45–53μm in diameter,into the common carotid artery.Six hours after modeling,fluorescent microspheres were observed directly through a fluorescence stereomicroscope,both on the brain surface and in brain sections.Changes in blood vessels,neurons and glial cells associated with microinfarcts were examined using fluorescence histochemistry and immunohistochemistry.The microspheres were distributed mainly in the cerebral cortex,striatum and hippocampus ipsilateral to the side of injection.Microinfarcts were found in the brain regions where the fluorescent microspheres were present.Here the lodged microspheres induced vascular and neuronal injury and the activation of astroglia and microglia.These histopathological changes indicate that this animal model of multiple cerebral infarctions effectively simulates the changes of various cell types observed in multifocal microinfarcts.This model is an effective,additional tool to study the pathogenesis of ischemic stroke and could be used to evaluate therapeutic interventions.This study was approved by the Animal Ethics Committee of the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences(approval No.D2021-03-16-1)on March 16,2021.

Enzymatic Histochemistry of Retina with Experimental Intraocular Pressure Elevation in Rabbits

To investigate the pathogenesis of retina lesions caused by intraocular pressure elevation, activities and distribution of enzymes in retina including lactic dehydrogenase (LDH), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), acid phosphatase (ACP), cholinesterase (ChE), cytochrome oxidase(CCO),nucleotidase (5'-Nase) and glucose-6-phosphatase (G6Pase) were determined histochemically in 30 rabbits. It was found that 1) in the early stage of intraocular pressure elevation, the activities of LDH, SDH, ATPase, ACP, and ChE in retina were increased, while the activities of CCO,5'-Nase decreased;2)in thelate stage of intraocular pressure elevation, the activities of all these enzymes but ACP, which showed a reduced activity, were close to the normal level; 3) in superoxide dismutase.(SOD-CCE) treated group, except the slight increase of LDH and G6Pase activities,the activities of the remaining enzymes were near to normal. Our results suggest that the various histochemical changes in retina induced by intraocular pressure elevation were cornpensatory in the early stage and were beneficial to the supply of energy needed in retinal tissue andcellular metabolism;while in the late stage, the lesion of retina cells developed due to decompensation.SOD-CCE could alleviate the retinal lesions caused by intraocular pressure elevation, and can be used as auxiliary drug for the treatment of intraocular pressure elevation.

ACETYLCHOLINESTERASE HISTOCHEMISTRY OF THE THALAMUS IN THE PRIMATE

Objective To observe the distribution of acetylcholinesterase activity in the thalamus of the monkey.Methods Histochemical method was used to detect the acetylcholinesterase activity in the thalamus.Results Acetylcholinesterase was found to be inhomogeneous distribution in the primate thalamus and to reveal previously uncovered inhomogeneity within certain thalamic nuclei and their subdivisions. The medial, ventral and posterior nuclear groups displayed markedly uneven acetylcholinesterase reaction.In the mediodorsal nucleus,three distinct sbudivisions were revealed by acetylcholinesterase histochemistry, medial magnocellular part, ventral sector of central parvicellular part and dorsolateral sector of lateral pars multiformity showed weak, moderate and strong acetylcholinesterase activity, respectively. In the ventral nuclear group, acetylcholinesterase histochemistry was strong in the medial part of ventral posterior nucleus, moderate in the magnocellular part of ventral anterior, caudal, medial, oral and pars postrema parts of ventral lateral nucleus, as well as lateral part of ventral posterior nucleus, poor and weak in the inferior part of ventral posterior nucleus, par compacta of the medial part of ventral posterior nucleus and parvicellular part of ventral anterior nucleus. In the pulvinar nucleus, acetylcholinesterase reaction ranged from weak, moderate to strong in the parts of the oral, medial and lateral, as well as inferior of this nucleus, respectively. Regional variations of acetylcholinesterase activity within the thalamic nuclei and their subdivisions can help to identify them by acetylcholinesterase histochemistry. In addition, the dark patches of strong acetylcholinesterase activity contrasting with a lighter surrounding matrix were revealed within the parvicellular part and pars multiformis of mediodorsal nucleus, paracentral nucleus, central lateral nucleus, pars postrema part of ventral lateral nucleus and medial habenula nucleus, as well as medial part of pulvinar necleus, respectively.Conclusion Acetylcholinesterase histochemistry can serve as useful marker for delineating the thalamic nuclei and their sbudivisions. The different distribution of acetylcholinesterase histochemistry implies the activity of thalamic nuclei may be modulated by cholinergic systems.

The 12th China-Japan Joint Seminar on Histochemistry and Cytochemistry

01 Differentiation-dependent changes in DNA methylation and histone H3 acetylation: possible roles of these epigenetic factors in mouse spermatogenesis Takehiko Koji Department of Histology and Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8523, Japan Mammalian spermatogenesis is a well-ordered process, consisting of spermatogonial mitosis, spermatocytic meiosis and spermiogenesis.The process is also characterized by successive chromatin condensation and a high frequency of spermatogenic cell apoptosis.Therefore, epigenetic factors can be implicated in the regulation of spermatogenesis and any epigenetic disorder may lead to the induction of apoptosis. When we analyzed global level of DNA methylation by immunohistochemistry with anti-5-methylcytosine in paraffin-embedded sections of mouse testis, there was no difference among any stages of germ cells.

STUDIES OF NUCLEOLAR ORGANIZER REGIONS AND MUCIN HISTOCHEMISTRY IN ULCERATIVE COLITIS

The argyrophil method for nucleolar organizer regions (AgNOR) known as the AgNor technique and mucin histochemical stain were applied to Investigate the dysplasia and cancaration in ulcerative colitis Including 58 cases of biopsy specimens and three cases operative specimens. The numbers of AgNOR have been gradully increased with the grades of dasplasls. Similarly the percentage of sialomucin having vicinal diols on side chain of sialic acid also gradully Increased with the grades of dysplasia. The AgNOR reflects rDNA transcriptional activity responsible for degrees of differentiation of cell. Epithelial cells secreting a heterogeneous mucin, could be taken as a signal of abnormal cellular differentiation. AgNOR and mucln chages might be assumed as a criteria In representing malignant transformation.

龙葵腺毛发育的细胞学研究

利用光学显微镜、扫描电镜和透射电镜技术,观察龙葵(Solanum nigrum L.)四叶一心期叶片及茎表皮腺毛的种类、分布,探究了不同类型腺毛的起源、生长、成熟、分泌、衰老等发育过程的细胞学特征;通过组织化学染色和荧光显微技术,观察了龙葵腺毛成分、分布,为龙葵的进一步开发利用提供参考。结果表明,(1)龙葵腺毛分为单细胞头腺毛和多细胞头腺毛两类,前者主要分布于茎表面和叶上下表皮,后者主要分布于茎表面的单细胞头腺毛之间、叶脉及叶边缘;(2)龙葵腺毛发育起始于表皮细胞突起,单细胞头腺毛顶端生长,具1~4个柄细胞,4种类型;多细胞头腺毛可再分为1层、2层与3层多细胞头腺毛,另具3种特殊类型;(3)龙葵成熟腺毛具分泌能力,通过皮下空间的物质积累导致腺毛头细胞表面形成突起、包块、破口,最终释放分泌物;而头细胞与柄细胞随即皱缩、衰老。(4)超微结构显示,腺毛头细胞中内质网与高尔基体极为丰富,合成代谢及分泌活动活跃,产生大量包裹嗜锇物质的囊泡,囊泡与细胞壁融合,进而将嗜锇物质转移至细胞壁并积累,随后储存在角质层下的皮下空间直至分泌释放;(5)组织化学染色结果表明,腺毛含有萜类、生物碱、脂类、蛋白质、酚类和多糖,头细胞中主要含有萜类、生物碱、脂类、蛋白质、酚类和中性多糖,柄细胞中主要含有萜类、生物碱、脂类。

家鸽消化道6种酶的分布与组织定位

目的研究家鸽消化道黏膜酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、腺苷三磷酸酶(ATPase)、非特异性酯酶(NSE)、琥珀酸脱氢酶(SDH)和过氧化物酶(POX)等6种酶的分布及组织定位。方法在成体家鸽消化道中的8个部位取样,采用冷冻切片、酶组织化学染色和光密度定量分析技术检测ACP、ALP、ATPase、NSE、SDH和POX的分布及活力。结果ACP在肌胃、空肠和十二指肠酶活力较高,而在腺胃、回肠、盲肠、直肠和食道酶活力较低。ALP在腺胃、十二指肠、空肠、回肠和直肠酶活力显著较高,盲肠和食道次之,肌胃酶活力最低。ATPase在除直肠外的其它部位酶活力均显著较高。NSE在肌胃酶活力最高,空肠、盲肠、回肠和食道次之,十二指肠、腺胃和直肠酶活力较低。SDH在食道、腺胃和肌胃酶活力显著较高,直肠次之,回肠、盲肠和空肠相对较低,十二指肠酶活力最低。POX在腺胃酶活力最高,肌胃、十二指肠、食道和回肠次之,直肠、空肠和盲肠酶活力较低。结论家鸽消化道6种酶的分布与其不同部位的消化吸收功能相适应。十二指肠和空肠是进行蛋白质细胞内消化和吸收的主要部位;小肠各部位对脂类、葡萄糖、钙离子以及无机磷酸盐均有较强的吸收作用;空肠、回肠和盲肠是消化酯类物质的主要部位。

山茱萸水提取物对肾阳虚动物模型肝脏和睾丸影响的实验研究

目的 :探讨山茱萸补肾阳作用。方法 :应用氢化考的松所致的肾阳虚动物模型 ,观察山茱萸水溶物对肾阳虚大鼠的影响。结果 :山茱萸水溶物具有使肾阳虚大鼠肝脏重量接近正常 ,肝细胞病理改变减轻 ,肝细胞RNA和糖原含量升高 ,丙二醛含量下降 ;它还能使肾阳虚大鼠睾丸脏器系数向正常方向逆转 ,使睾丸间质细胞内RNA含量增加。结论

一氧化氮合酶阳性神经元在小鼠脑内的分布

目的 :研究一氧化氮合酶 (NOS)在小鼠脑内的分布。方法 :用NADPH 黄递酶组织化学技术 ,观察了NOS阳性神经元在小鼠脑内的分布和形态。结果 :在大脑皮质、纹状体、基底前脑、杏仁核、下丘脑和脑干等处有较多一氧化氮合酶阳性神经元的分布。结论

退变性颈椎失稳颈后深部肌肉的组织化学特征及其临床意义

目的：研究颈后深部肌肉的肌纤维型分布和横切面积，探讨其生理功能和在退变性颈椎失稳发病中的作用。方法：应用肌球蛋白ＡＴＰ酶染色法，观察正常人（４例）和退变性颈椎失稳患者（４例）颈后深部肌肉各型肌纤维的比例和结构特征，并测量肌纤维的横切面积。结果：正常人和退变性颈椎失稳患者颈后深部肌肉Ⅰ型和Ⅱ型纤维交织排列，横切面呈多边形或椭圆形，Ⅰ型纤维数量及横切面积均显著大于Ⅱ型纤维。退变性颈椎失稳患者肌纤维边界不清，横切面上可见小空泡，Ⅰ型纤维减少，Ⅱ型纤维增多。结论：颈后深部肌肉对维持颈段脊柱的生理姿势和运动起重要作用，肌纤维病理学改变是退变性颈椎失稳发病的重要因素之一。

先天性马蹄内翻足小腿肌肉多项组织化学研究

本文对13例先天性马蹄内翻足及5例非神经肌肉疾病手术患儿小腿71个肌肉活检标本进行了多项组织化学研究。肌纤维ATP酶染色,发现先天性马蹄内翻足小腿胫后肌及腓肠肌Ⅰ型肌纤维增多及同型肌纤维聚集现象,提示有肌纤维失神经后神经再支配;肌纤维吖啶橙荧光染色及运动终板AchE染色未发现肌纤维有失神经后强橙黄色荧光及运动终板形态、功能异常,说明胚胎在生长发育过程中通过正常的神经再支配,达到了神经与肌肉在结构和功能上的完整。本实验研究结果支持早期肌力不平衡学说,为早期肌力平衡手术治疗提供了理论依据。

水貂发情期卵巢动态结构研究

对发情期水貂卵巢进行系列切片，研究其卵泡生长．闭锁、排卵．间质腺细胞的形态结构及组织化学变化。水貂卵泡生长呈波浪式，卵泡闭锁分３种类型、闭锁有腔卵泡内膜中上皮样细胞增殖、在原有卵泡腔的位置上形成新的间质腺组织。并对含有甘油三酯及磷脂的脂滴分布以及涉及到类固醇生物合成等方面的问题进行了讨论。