自发性高血压大鼠下丘脑加压素能神经元形态学研究

本文用光、电镜和免疫组织化学方法观察了自发性高血压大鼠（ＳＨＲｓ）下丘脑加压素能神经元的形态结构。电镜下ＳＨＲｓ加压素能神经元胞体大,电子密度高,核大有皱褶细胞质内密集充满各种细胞器。可见膜包神经分泌颗粒。ＰＡＰ法显示精氨酸加压素（ＡＶＰ）阳性细胞主要分布于视上核（ＳＯＮ）腹外侧部及室旁核（ＰＶＮ）内。在ＳＯＮ和ＰＶＮ之间尚有一个ＡＶＰ阳性细胞的密集核团。说明ＳＨＲ下丘脑也有产生ＡＶＰ功能。ＳＯＮ和ＰＶＮ之间的核团是否与ＳＨＲｓ下丘脑－垂体－肾上腺轴ＡＣＴＨ的异常有关？有待进一步研究。本文为探讨ＳＨＲｓ正常生理功能及高血压的发生提供形态学资料。     

大鼠实验性脾虚证胰腺组织化学研究

用成年雄性Ｗｉｓｔａｒ大鼠３０只,分为（１）正常对照组,喂饲自来水。（２）脾虚组,用苦降破气中药和饮食失节法致成脾虚模型。（３）自然恢复组,动物致虚后,喂饲自来水。（４）中药治疗组。取四组动物胰腺进行ＲＮＡ,琥珀酸脱氢酶（ＳＤＨ）,乳酸脱氢酶（ＬＤＨ）,三磷酸腺苷酶（ＡＴＰａｓｅ）,葡萄糖－６－磷酸酶（Ｇ－６－Ｐａｓｅ）和硫胺素焦磷酸酶（ＴＰＰａｓｅ）组织化学反应和观察,并对ＳＤＨ,ＬＤＨ,ＲＮＡ进行了显微分光光度计定量测定。本研究结果表明,脾虚组胰腺泡细胞的ＲＮＡ,ＳＤＨ,ＡＴＰａｓｅ,Ｇ－６－Ｐａｓｅ,ＴＰＰａｓｅ含量和活性都低于对照组,而ＬＤＨ活性高于对照组。治疗组与自然恢复组相比,治疗组胰腺泡细胞以上指标接近对照组。定量测定与定性的结果一致。本研究表明,脾虚证时胰腺泡细胞上述几种酶活性和ＲＮＡ明显下降,可能在脾虚证发病中起主要作用,中药治疗有显著改善     

国人睾丸、附睾和输精管的水解酶的组织化学观察

本文报导国人睾丸、附睾和输精管的ＮＳＥ,ＡＣｈＥ,ＣｈＥ,ＡＬＰ,ＡＣＰ,５＇－Ｎａｓｅ,Ｇ－６－Ｐａｓｅ,β－ＧＡ,β－ＧＲ,ＡＰ－Ｍ,ＡＴＰａｓｅ和ＴＰＰａｓｅ水解酶的组织化学活性、结果显示：睾丸曲细精管的ＡＬＰ,５＇－Ｎａｓｅ和ＡＴＰａｓｅ;睾丸间质细胞的ＮＳＥ,ＡＬＰ,ＡＴＰａｓｅ和ＡＣＰ;睾丸间质中的ＮＳＥ,ＡＬＰ和ＡＴ－Ｐａｓｃ;睾丸输出小管和附睾管上皮的ＮＳＥ,ＡＬＰ,ＡＣＰ,５’－Ｎａｓｅ,β－ＧＡ,β－ＧＲ,ＡＴＰａｓｅ和Ｔｐｐａｓｅ;附睾头部间质中的ＮＳＥ,ＡＣｈＥ,ＡＬＰ,和ＡＴＰａｓｅ;输精管上皮细胞的ＡＴＰａｓｅ的酶活性均呈强阳性或极强阳性。说明人类睾丸、附睾和输精管含有丰富的水解酶,尤其是附睾头部的输出小管、附睾管和头质均含有种类多活性高的水解酶,在精子的功能成熟上起了重要的作用,提示其可能作为生育与不育诊治中的重要指标。     

颗粒子宫腺细胞的分离、纯化与细胞化学研究

胰蛋白酶消化小鼠子宫腺细胞区,制成单细胞悬液,滴片后进行细胞学及细胞化学研究。结果：有多种类型细胞存在于悬液中,其中中等大小细胞（直径２０μｍ左右）大部分为颗粒子宫腺细胞（ＧｒａｎｕｌａｔｅｄＭｅｔｒｉａｌＧｌａｎｄＣｅｌｌｓ,ＧＭＧｃｅｌｌｓ）。ＧＭＧ细胞约占所有细胞的５０％。利用Ｐｅｒｃｏｌｌ非连续性密度梯度离心可以提高ＧＭＧ细胞的纯度,使之达８０％以上。细胞化学研究显示ＧＭＧ细胞含ＡＣＰ,ＮＳＥ,ＬＮＡｓｅ及糖蛋白成份。     

用原位末端标记方法观察细胞凋亡在正常喉粘膜、炎症、癌前和癌变中的作用

为了观察凋亡与喉粘膜癌变的关系,本实验应用原位末端标记（ＩＳＥＬ）方法检测喉粘膜正常（ｎｏｒｍａｌｌａｒｙｎｇｅａｌｍｕｃｏｓａ,Ｎ）、炎症（ｉｎｆｌａｍｍａｔｉｏｎ,ＩＦ）、不典型增生（ｄｙｓｐｌａｓｉａ,ＤＹＳ）及鳞癌（ｓｑｕａｍｏｕｓｃｅｌｃａｒｃｉｎｏｍａ,ＳＣＣ）中细胞凋亡情况。标记后的凋亡细胞通过计数以凋亡指数（ＡＩ）表示。结果表明：ＤＹＳ组的凋亡指数（ＡＩ）在所有病变中最高;ＤＹＳ及ＳＣＣ组的ＡＩ值较正常Ｎ及ＩＦ组明显增多,且有显著性差异（Ｐ＜００５）;对ＳＣＣ的计数显示,ＡＩ值随着ＳＣＣ的分化程度下降有逐渐减少的趋势。ＳＣＣ转移组与非转移组的ＡＩ值间无显著性差异（Ｐ＞００５）。结论：细胞凋亡在喉粘膜癌前病变和癌变中发挥着重要作用,与喉癌的发生发展密切相关     

中药固真方对促甲状腺素受体基因表达的调节作用

促甲状腺素（ＴＳＨ）通过甲状腺细胞膜上的ＴＳＨ受体（ＴＳＨＲ）,产生第二信使ｃＡＭＰ,从而激活ｃＡＭＰ反应性起动子,而使相应的基因获得表达．实验结果表明,在构建有ＴＳＨＲ和糖蛋白激素ｃＡＭＰ反应性起动子以及萤光素基因（Ｌｕｃ）的转染细胞中,经补肾益精中药固真方提取液（１×１０~（－４）稀释）处理３～５ｄ后,可下调ＴＳＨＲ基因的表达,并使ＴＳＨＲ数目减少．提示固真方可调整甲状腺细胞的功能,或许有利于调整甲状腺机能亢进．     

大肠癌及癌旁粘膜中胃泌素、生长抑素细胞的免疫细胞化学研究

本文对１７例大肠癌病人癌组织及癌旁粘膜中的胃泌素细胞和ＳＳ细胞进行免疫细胞化学观察（ＰＡＰ）。结果显示：在癌及癌旁粘膜中来见胃泌素细胞;癌细胞中均未见ＳＳ细胞;癌旁粘膜中ＳＳ细胞的形态、位置近于正常。距癌灶０～２ｃｍ粘膜ＳＳ细胞数和总阳性强度显著高于２～６ｃｍ和４～６ｃｍ粘膜（Ｐ＜０．０１）。０～２ｃｍ粘膜中ＳＳ细胞数和总阳性强度与粘膜上皮细胞异型增生程度呈显著负相关（前者Ｐ＜０．０１;后者Ｐ＜０．０５）。Ｄｕｋｅｓ’ＣＤ期者０～２ｃｍ和２～４ｃｍ粘膜中细胞平均阳性强度均显著低于ＡＢ期患者（前者Ｐ＜０．０５,后者Ｐ＜０．０１）。距癌０～２ｃｍ粘膜ＳＳ细胞增多和总阳性强度增高可能是机体发生在病变部位的胃肠内分泌防御反应的细胞学基础,并受到粘膜上皮异型增生程度的影响。晚期０～４ｃｍ粘膜ＳＳ细胞分泌ＳＳ减少是机体胃肠内分泌功能衰退的表现。     

PAF对肺内小动脉平滑肌细胞TXA_2，PGI_2乃Ca~（2＋）－ATPase的影响

本工作采用分离培养家兔肺内小动脉平滑肌细胞（ＰＡＳＭＣｓ），观察了外源性血小板活化因子（ｐｌａｔｅｌｅｔａｃｔｉｖａｔｉｎｇｆａｃｔｏｒ，ＰＡＦ）、ＢＮ５２０２１（ＰＡＦ受体拮抗剂）、吲哚美辛、维拉帕米对ＰＡＳＭＣｓ产生血栓素Ａ_２（ＴｘＡ_２）、前列环素（ＰＧＩ_２）及对细胞膜Ｃａ~（２＋）－ＡＴＰａｓｅ活力的影响。结果表明：（１）基础状态下ＰＡＳＭＣｓ存在花生四烯酸（ＡＡ）代谢。（２）外源性ＰＡＦ通过受体后途径激活环加氧酶促进ＡＡ代谢致ＴＸＡ_２及ＰＧＩ－２增加，ＴＸＡ_２／ＰＧＩ_２比值无明显变化。（３）外源性ＰＡＦ能直接抑制Ｃａ~（２＋）－ＡＴＰａｓｅ活力。（４）维拉帕米可逆转ＰＡＦ抑制ＰＡＳＭＣｓ膜Ｃａ~（２＋）－ＡＴＰａｓｅ活力的效应。     

门静脉分支结扎后肝的组织化学变化

为了进一步探讨门静脉血流阻断对肝的影响,作如下实验。２４只健康成年家兔分为（１）对照组：假手术;（２）实验组：门静脉右外支结扎术。术后第１,３,７天分别处死动物。取结扎叶肝组织行组织化学观察。结果,实验组肝琥珀酸脱氢酶（ＳＤＨ）,葡萄糖６磷酸酶（Ｇ６Ｐａｓｅ）,镁激活三磷酸腺苷酶（Ｍｇ２＋ＡＴＰａｓｅ）,胆碱酯酶（ＣｈＥ）等酶活性和糖原ＰＡＳ反应明显降低;乳酸脱氢酶（ＬＤＨ）,酸性磷酸酶（ＡＣＰ）和碱性磷酸酶（ＡＬＰ）活性明显升高。术后第１,３,７天各动物肝的组织化学变化无显著性差异。结果表明：门静脉缺血将引起肝的代谢和功能损害,门静脉在肝血供中起着很重要的作用     

缺氧时肺动脉内皮细胞与肺动脉平滑肌细胞增殖的相互影响

血管内皮细胞和血管平滑细胞在结构和功能上关系密切,两者的相互在与血管舒缩笔血和壁结构。本文观察了培养的小牛肺动脉内皮细胞（ＰＡＥＣｓ）和肺动平滑肌细胞（ＰＡＳＭＣＳ）缺氧时在细胞增殖方面的相互影响。ＰＡＳＭＣＳ常氧条件培养基（ＣＭ）可使ＰＡＥＣＳ的＾３Ｈ－ＴｄＲ掺入降低约５８％,缺氧ＣＭ对ＰＡＥＣＳ的＾３Ｈ－ＴｄＲ掺入无明显的抑制作用;ＰＡＥＣＳ的常氧ＣＭ使ＰＡＳＭＳ的＾３Ｈ－ＴｄＲ掺入升高约６０     

不同周龄SD大鼠脏器重量及其变化趋势分析

目的分析不同周龄SD大鼠的脏器重量及其变化趋势,为评判药物毒性反应提供理论参考。方法分别选取试验第13、26、52、78和104周对照组动物脑、脾脏、心脏、肺脏、肝脏、肾脏、肾上腺、睾丸、卵巢的重量数据并分析。结果从13～104周SD雌鼠脑、脾脏、心脏、肺脏、肝脏、肾脏、肾上腺、卵巢的重量呈升高趋势。从13～104周SD雄鼠脑、脾脏、心脏、肺脏、肝脏、肾脏重量均重于雌鼠,但雌鼠肾上腺重量、脏体比和脏脑比均显著高于雄鼠。结论本研究首次在国内建立了符合我国实验动物现状的,不同周龄SD大鼠的脏器重量背景数据和参考值范围,并分析了不同周龄SD大鼠脏器重量变化趋势。     

实验性脾虚证大鼠结肠和盲肠组织化学、免疫组织化学研究

成年雄性Wistar大鼠60只,分正常对照组;脾虚组;自然恢复组和四君子汤治疗组。四组动物取结肠和盲肠分别进行H-E;酶组织化学和5-羟色胺细胞PAP免疫组织化学观察。结果表明,脾虚组结肠和盲肠粘膜上皮的杯状细胞较对照组明显增多,淋巴细胞及淋巴集结也增多。固有层毛细血管充血并有炎症细胞的增多。上皮细胞和腺细胞琥珀酸脱氢酶(SDH)减弱,乳酸脱氢酶(LDH)增强;结肠上皮细胞和腺细胞碱性磷酸酶(AIP)和三磷酸腺苷酶(ATPase)增强,而盲肠上皮细胞和腺细胞AIP和ATPase减弱。结肠和盲肠5-HT细胞免疫组化反应减弱。这些结果表明肠上皮细胞酶的变化和5-HT细胞分泌活性的变化与脾虚证的发生密切相关,可能是导致脾虚证原因之一。经四君子汤治疗后以上各项指标均比自然恢复组更接近于正常对照组,说明此药对消化道粘膜上皮酶活性和5-HT细胞分泌活性恢复正常有明显效果,从而起治疗作用。     

Tissue manganese levels and liver pyruvate carboxylase activity in magnesium-deficient rats

To investigate the manganese status in magnesium deficiency, 40 male Wistar rats, 3 wk old, were divided into two groups and fed a magnesium deficient diet or a normal synthetic diet for 2 wk. Dietary magnesium depletion decreased magnesium levels in brain, spinal cord, lung, spleen, kidney, testis, bone, blood, and plasma, while it elevated the magnesium level in liver. In magnesium-depleted rats, calcium concentration was increased in lung, liver, spleen, kidney, and testis, while it was decreased in tibia. In magnesium-depleted rats, manganese concentration was decreased in plasma and all tissues except adrenal glands and blood. Dietary magnesium depletion diminished pyruvate carboxylase (EC 6.4.1.1) activity in the crude mitochondrial fraction of liver. Positive correlation was found between the liver manganese concentration and the pyruvate carboxylase activity. In the magnesium-depleted rats, glucose was decreased while plasma lipids (triglycerides, phospholipids, and total cholesterol) were increased. These results suggest that dietary magnesium deficiency changes manganese metabolism in rats.     

脾虚证小鼠胸腺细胞周期和免疫细胞化学的实验研究

本文用ＦＣＭ和免疫细胞化学技术检测了小鼠胸腺细胞的活动周期,Ｂｒｄｕ＋细胞和表达Ｔｈｙｌ．２抗原的Ｔ细胞。小鼠包括对照组,利血平诱发的脾虚组,自然恢复组和健脾汤治疗后复健组。ＦＣＭ和免疫细胞化学结果显示,脾虚组小鼠Ｇ１期细胞堆积,Ｓ期和Ｇ２＋Ｍ期细胞减少。同时胸腺内Ｂｒｄｕ＋细胞和Ｔ细胞也相应减少。经过健脾汤治疗后,除Ｇ１期细胞外,Ｓ期和Ｇ２＋Ｍ期细胞,Ｂｒｄｕ＋细胞和Ｔ细胞均显著增加,并且几乎达正常水平。但是自然恢复组在恢复细胞活动周期正常化,以及恢复Ｂｒｄｕ＋细胞和Ｔ细胞方面比复健组缓慢。说明健脾汤对脾虚小鼠的胸腺细胞有促进ＤＮＡ合成和细胞增殖功能。     

Perspectives in Endocrine Toxicity of Heavy Metals—A Review

An attempt has been made to review the endocrine/hormonal implications of a few environmentally significant metals, viz, lead, mercury, cadmium, copper, arsenic and nickel, in man and animals. Special emphasis has been given to the adrenals, thyroid, testis, ovary and pancreas. Toxic metals can cause structural and functional changes in the adrenal glands. Their effects on steroidogenesis have been reviewed. It has been reported that thyroid hormone kinetics are affected by a number of metallic compounds. Occupational exposure to a few of these metals can cause testicular injury and sex hormone disturbances. Protective effects of a few antioxidants on their reproductive toxicity have also been discussed. Information gathered on female reproductive toxicity of heavy metals shows that exposure to these metals can lead to disturbances in reproductive performance in exposed subjects. Certain metals can cause injury to the endocrine pancreas. Exposure to them can cause diabetes mellitus and disturb insulin homeostasis. The need to develop molecular markers of endocrine toxicity of heavy metals has been suggested. Overall information described in this review is expected to be helpful in planning future studies on endocrine toxicity of heavy metals.     

The enhancement of body resistance to combined exposure to immobilization and cold with thyroid hormones]

In experiments on 123 male rats have been found that the combination of immobilizing and cold (4 degrees C) stresses during 6 hours results in the formation of 4.5 gastric mucosa ulcers in every rat, marked activation of lipoperoxidation and depression of antioxidant activity of myocardium, significant increase of relative mass of adrenal glands, decrease of spleen mass and death of 80% animals accompanied by the reduction of concentration of thyroid hormones in blood serum and hypothermia (to 28 degrees C). The prestress injection of small doses of thyroidine decreased of mucosa defect rate and the number of ulcers to 2.8, made less marked the changes of relative mass of adrenal glands and spleen and the changes of concentration of thyroid hormones in blood serum, significantly limited the activation of lipoperoxidation, depression of power in antioxidant systems of myocardium, hypothermia (to 32.5 degrees C) and significantly increased the survival rate (to 70%).     

Period gene expression in mouse endocrine tissues

Circadian rhythms are generated by the oscillating expression of the Per1 and Per2 genes, which are expressed not only in the central brain pacemaker but also in peripheral tissues. Hormones are likely to coordinate physiological function in time. We performed in situ hybridization to localize mPer1 and mPer2 mRNA to particular cell types and tissue compartments in adrenal, thyroid, and testis. BALB/c mice maintained in a 12:12-h light-dark cycle expressed mPer1 in adrenal medulla, particularly in late afternoon and early night. mPer2 mRNA was more intensely expressed in adrenal cortex, especially in afternoon and evening. mPer1 mRNA was detected in thyroid. mPer1 was found in some but not all seminiferous tubules of each mouse at all times of day. Quantitation in C57BL/6 mice revealed a significant increase in the number of heavily labeled seminiferous tubules early in the night. Consistent with in situ hybridization, immunocytochemistry showed PER1 protein in spermatocytes and spermatids (spermatogenic stages VII-XII). Staining in spermatogonia and interstitial cells was inconsistent. Double labeling with 5'-bromodeoxyuridine showed PER1 expression first occurring 5 days after DNA replication. We conclude that mPeriod genes are expressed in peripheral endocrine glands. Central regulation, adenohypophyseal control, and functional importance of expression and phase remain to be elucidated.     

The effect of a deficiency of the hormones of the peripheral endocrine glands on behavioral, learning and memory processes]

A comparative study of the effects of thyroid, adrenal, and gonadal deficit on capability for learning, retention of memory traces, and behavior was carried out in male rats under conditions of hormonal disbalance produced by extirpation of the endocrine glands. Behavior of animals was tested during the active and passive avoidance learning and in the open field. It was found out that the extirpation of the peripheral endocrine glands impairs learning and reproduction of the acquired reaction and alters the behavior. The results suggest that corticosteroid hormones take part in learning and behavior. Gonadal and thyroid hormones appear to exert a modulating influence on the higher nervous activity.     

Subcellular location and expression pattern of autoimmune regulator (Aire), the mouse orthologue for human gene defective in autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED).

Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED), also known as autoimmune polyglandular syndrome Type I (APS1), is an autosomal recessive autoimmune disease caused by mutations in a gene designated as AIRE (autoimmune regulator). Here we have studied the expression of Aire in transfected cell lines and in adult mouse tissues. Our results show that Aire has a dual subcellular location and that it is expressed in multiple immunologically relevant tissues such as the thymus, spleen, lymph nodes, and bone marrow. In addition, Aire expression was detected in various other tissues such as kidney, testis, adrenal glands, liver, and ovary. These findings suggest that APECED protein might also have a function(s) outside the immune system.(J Histochem Cytochem 49:197-208, 2001)     

Morphological and functional changes in several endocrine glands induced by hypothyroidism in the rat

The effect of hypothyroidism upon the morphology and the function of several endocrine glands was studied in radiothyroidectomized male rats. It was found that T3, T4, insulin, prolactin and corticosterone levels were significantly lower in hypothyroid rats. TSH levels were significantly higher in these animals while no changes were depicted in testosterone levels. The administration of T4 drew back to normal range the above-mentioned altered serum hormone levels. The studies performed with light microscopy revealed alterations only in the TSH secretory cells of the adenohypophysis. Conversely, when using the electron microscope to study the different endocrine glands, clear alterations were depicted in the TSH and prolactin secretory cells of the adenohypophysis, as well as in the pancreatic B cells and the cells of the zona fasciculata of the adrenal cortex. No abnormal changes were demonstrable at the level of the seminiferous tubules of the testis. All the above morphological changes were corrected by the administration of T4 to hypothyroid rats. These results suggest that the hypothyroid state is a complex hormonal dysfunction rather than a single hormonal defect. The secretory alterations are accompanied by fine cellular alterations in the corresponding glands.