改良NADPH—d和AchE酶组织化学法诊断先天性巨结肠

目的：探讨进一步改良后的还原型尼克酰胺腺嘌呤二核苷酸黄递酶（NADPH—d）和乙酰胆碱酯酶（AchE）酶组织化学染色法诊断先天性巨结肠（HD）的优势,以期提高HD术前及术中诊断的准确率。方法：取HD患儿术中切除的结肠标本18例,其中女7例,男11例,平均年龄（8．6±3）月。标本分狭窄段、移行段、扩张段,其冰冻切片分别用改良的NADFH—d和AchE酶组织化学染色法进行染色,与常规HE染色进行比较,并用肠神经元特异标记物β-Ⅲ Tubulin（TUJ-1）行免疫荧光及蛋白基因产物9．5（proteingeneproduct9．5,PGP9．5）行免疫组化检查论证改良方法的特异性。结果：NADPH—d法能快速确定扩张段近端切缘是否存在神经节细胞,诊断HD的阳性率为94．4％（17／18）;AchE法对于HD狭窄段黏膜下异常增生的神经丛诊断阳性率为88．9%（16／18）;两组间诊断阳性率差异无统计学意义（P〉0．05）。耗时方面NADPH-d法为（7．0±1．7）min,AchE法为（30．9±2．2）min,组间比较差异有统计学意义（P〈0．001）。上述两种改良方法与TUJ-1及PGP9．5对照结果一致。结论：NADPH—d法能快速确定肠管切缘是否有神经节细胞分布,适用于术中快速诊断HD;AchE法适用于术前直肠黏膜层的活检诊断。     

Histochemical and pharmacological analysis of catecholaminergic projections to the perifornical hypothalamus in relation to feeding inhibition

Three techniques, namely, midbrain lesions, fluorescence histochemistry, and brain cannulation, were used in combination to analyze noradrenergic projections to the paraventricular nucleus of the hypothalamus (PVN) and their function in stimulating feeding behavior. The convergence of evidence indicates that the dorsal component of the central tegmental tract (CTT), which ascends through the dorsal pons and then projects through the medial tegmental radiations (TR) into the ventral tegmentum just dorsal to the medial lemniscus, contains the crucial noradrenergic axons which innervate the PVN and mediate noradrenergic stimulation of feeding behavior. The primary evidence for this conclusion is that dorsal tegmental electrolytic or 6-OHDA lesions which damaged specifically these fibers invariably caused: (1) a reduction of catecholamine varicosities within the PVN (most notably, fine and moderate-size, rounded varicosities within the parvocellular area); (2) a strong reduction or loss of the feeding response elicited by PVN injection of the presynaptically-acting drugs tranylcypromine and desipramine; and (3) a potentiation of the same response produced by injected norepinephrine. These pharmacological and neurochemical changes in the PVN were reduced in magnitude if the dorsal CTT and medial TR fivers received only partial damage, and these changes did not occur at all if the lesion fell immediately dorsal to these fibers without damaging them. Specific lesions in the ventral tegmentum, which also failed to damage the dorsal CTT and TR axons but instead damaged the ventral component of the CTT, not only failed to disrupt the action of the antidepressant agents but actually potentiated their effectiveness in the PVN. Ventromedial lesions, however, which severed the rostroventral extension of the dorsal CTT and medial TR fibers, had the same behavioral consequences as had the dorsal lesions which damaged this projection at a more dorsocaudal level. Finally, damage to other catecholamine projections had little effect on PVN function in stimulating eating.     

Metabolic changes after injection of quinolinic acid or 6-hydroxydopamine in the rat striatum: A time-course study using cytochrome oxidase and glycogene phosphorylase a histochemistry

Abstract

Injection of excitotoxins, such as quinolinic acid (QA), into the striatum has been extensively used as an experimental model of Huntington's disease, while Injection of 6-hydroxydopamine (6-OHDA) into the dopaminergic nigrostriatal pathway provides a well established model of Parkinson's disease. In the present study, we have examined the metabolic changes induced by an intrastriatal injection of QA or 6-OHDA using histochemical staining for the metabolic markers cytochrome oxidase (COx) and active glycogene phosphorylase (CPa). Intrastriatal injection of QA produced major changes in COx (decrease of staining) and CPa (increase of staining, except in the core of the lesion where the staining was virtually absent) histochemistry at the level of the striatum and of most of the other basal ganglia nuclei. Although attenuated over time, these changes persisted up to one year after the lesion. On the contrary, after the intrastriatal injection of 6-OHDA (which induces only a partial lesion of the nigrostriatal pathway), we did not observe any remarkable changes in COx or CPa staining. This study illustrates the discrepancies between the morphological changes and metabolic changes that are induced when using these experimental models of neurodegenerative disorders. [Neurol Res 2000; 22: 425-429]     

Histochemical evaluation of energy metabolism in rat glioma

Abstract

The key enzymes of oxidative phosphorylation and glycolysis were evaluated histochemically in rat-implanted C6 gliomas using spot densitometry. Hexokinase, the initial enzyme for the glycolysis pathway, was 40% higher within tumour than the contralateral cerebral cortex. A similar increase within tumours for 2-deoxyglucose was observed by autoradiography. Glucose-6-phosphate dehydrogenase (G6PDH), which is the first enzyme in the pentose phosphate pathway, shunting glucose towards nucleic acid synthesis, was more than 300% higher in gliomas compared with the normal cortex. In contrast, enzymes in the energy producing tricarboxylic acid cycle (succinate-isocitrate-and malate-dehydrogenase) and in the electron-transport system (cytochrome c oxidase) were significantly reduced in tumour (58% less than the contralateral cortex). Lactate dehydrogenase activity, which converts pyruvate to lactate, was 50% higher within tumour. Significant reductions of enzymatic activities also occurred in non-neoplastic tissue in ipsilateral hemisphere, with larger tumours. Some enzymes showed heterogeneous activity within tumours, especially C6PDH. These results suggest that: (1) energy production is more dependent on lactate production than on oxidative phosphorylation in C6 gliomai, and (2) a significant part of the increased glucose utilization in glioma cells is due to increased activity of the pentose phosphate shunt for increased DNA synthesis, and not energy production. [Neurol Res 1992; 14: 289-293]     

真皮下血管网皮瓣血运重建方式的实验研究

目的探讨真皮下血管网皮瓣血运重建的机制。方法以SD大鼠为实验动物,在背部形成蒂在尾侧的2cm×7cm真皮下血管网皮瓣,并将皮瓣人为分成3个区段:1区为近蒂部0~3cm;2区为距蒂部3~5cm;3区为皮瓣远端距蒂部5~7cm。皮瓣直接原位缝合为对照组,分别在1、2区段下方放置硅胶膜为实验组,采用大体肉眼观察、碱性磷酸酶组织化学和免疫组化(CD34)的方法,比较各组皮瓣成活以及不同区段微血管构筑的差异。结果与对照组及1区下方放置硅膜相比,2区下方放置硅膜后皮瓣成活率显著下降;1区下方放置硅膜与对照组之间皮瓣成活率无统计学差异。对照组及在1区下方放置隔膜后皮瓣2区微血管密度较同组1区明显增高。结论真皮下血管网皮瓣中段的缺血缺氧及血运重建对超长比例皮瓣的成活尤其是对皮瓣远端的血运重建具有重要作用。     

雌激素对慢性脑缺血大鼠海马 CA1区一氧化氮合酶阳性神经元的影响

目的：研究雌激素对去势后慢性脑缺血大鼠海马CA1区一氧化氮合酶阳性神经元的影响。方法：15只大鼠随机分为模型组、(雌激素)治疗组、假手术组。利用卵巢摘除术和双侧颈总动脉永久结扎术制备动物模型，分别予以雌激素和煮沸过的麻油处理。B—NADPH组织化学染色显示一氧化氮合酶(NOS)阳性神经元，光镜下观察、分析。结果：模型组和治疗组海马CA1区NOS阳性神经元数目与假手术组相比均有明显减少，与治疗组相比，模型组减少更多。结论：慢性脑缺血能使去势大鼠海马CA1区NOS阳件神绎元数目明显减少．雌激素替代治疗能够减轻这种改变．     

水淹对钉螺卵组织化学影响的实验研究

１９９３年４～５月．采用实验现场结合组织化学技术，观察了水淹对钉螺春季繁殖器螺卵组织化学的影响。结果表明，对照组螺卵卵胚内的ＳＤＨ、ＬＤＨ及ＣＣｏ等与细胞能量代谢有关的酶呈强阳性反应．实验组螺卵卵胚内的ＳＤＨ．ＣＣｏ，水淹第１０ｄ时降为的阳性反应（＋），３０ｄ时消失，ＬＤＨ含量在水淹在２０ｄ时未见变化，３０ｄ时降为阳性反应（＋＋），４０ｄ时消失。水淹３０ｄ的螺卵石蜡包理切片，作糖原（ＰＡＳ）反应，由强阳性变为弱阳性（＋）．综合上述结果，分析了水淹影响螺卵发育的机制。     

Alteration in immunoexpression of glucose transporter 2 in liver of tumour-bearing rats

To elucidate interactions between the glucose transport system and hepatic glucose production in the tumour-bearing state, glycogen storage, expression of glucose transporter isoform 2 (Glut 2) and activities of glucose-6-phosphatase (G-6-Pase) and hexokinase were histochemically examined in hepatocytes of tumour-bearing rats. Five male F344 rats, subcutaneously inoculated with methylcholanthrene(MCA)-induced sarcoma cells were compared with five pair-fed animals and four ad libitum fed controls. Glycogen storage was markedly decreased in liver cells of tumour-bearing rats compared to in those of control animals. Glut 2 immunoreactivity was uniformly seen in the cellular membrane of hepatocytes from control animals. In rats bearing sarcoma, the staining intensity was significantly decreased, suggesting that Glut 2 with its bi-directional transport capacity was down-regulated in the tumour-bearing state. Positive staining for hexokinase activity was located in the perivenous area in livers from control animals and was more diffusely located and more intense in livers from tumour-bearing animals. G-6-Pase activity, limited to the peripheral area in livers from controls, extended to the intermediate area and had stronger reactivity in livers from tumour-bearing animals. In the tumour-bearing cachectic condition, glucose may be partially consumed by a futile cycle, hepatic metabolic zonation was disturbed, and the release of glucose from the liver may not be mediated by a facilitative glucose transporter-2.     

大鼠室周器官的形态学观察

目的 系统观察5个室周器官的微细结构。方法 对40只成年SD大鼠行脑切片，分别行硫堇和Tolivia还原银染色，以及NADPH-d组织化学染色。结果 （1）室周器官微小的体积与其异常丰富的血床形成鲜明的反差；（2）5个室周器官依体积大小次序为穹隆下器，最后区，正中隆起，连合下器，终板血管器；（3）穹隆下器和终板血管器可能含神经分泌细胞；（4）连合下器具有室管膜下神经细胞层；（5）正中隆起具有最密集和最多的伸展细胞。一氧化氮合酶均呈阳性；（6）在最后区与延髓中央管之间，可能有神经纤维联系。结论 室周器官可能是脑内体液化学信息的重要转导位点。