降钙素基因相关肽和神经生长因子对局灶性脑缺血再灌注大鼠顶叶、海马FasmRNA表达的影响

目的研究外源性降钙素基因相关肽(CGRP)和神经生长因子(NGF)对局灶性脑缺血再灌注大鼠顶叶、海马FasmRNA表达的影响,探讨降钙素基因相关肽和神经生长因子对缺血再灌注脑神经组织的作用。方法用线栓法制备大鼠大脑中动脉阻塞(MCAO)模型,应用原位杂交和显微图象分析方法检测局灶性脑缺血再灌注大鼠顶叶、海马FasmRNA的表达。结果假手术组大鼠顶叶、海马未见FasmRNA阳性表达细胞;缺血再灌注组(缺血再灌注不同时间段6h,12h,24h,48h,72h)顶叶、海马FasmRNA阳性细胞明显过表达;注射CGRP或NGF后顶叶、海马FasmRNA阳性表达细胞平均光密度值明显低于缺血再灌注组(P<0.01);两者合用组平均光密度值比单独应用低(P<0.05)。结论经颈动脉注入外源性CGRP和NGF下调顶叶和海马缺血神经元FasmRNA的表达,且有协同效应,可能是两者对缺血神经元保护作用的机制之一。     

甲状腺滤泡旁细胞的研究进展

甲状腺滤泡旁细胞是甲状腺中产生降钙素的细胞。但作为ＡＰＵＤ系的甲状腺滤泡旁细胞也有分泌多种调节肽的功能。本文对甲状腺滤泡旁细胞分泌调节肽的种类，分泌调节肽的基因机制以及调节肽的作用机制的研究进行综述     

人甲状腺促性腺激素释放激素及其受体的表达

目的　探讨人甲状腺中是否存在促性腺激素释放激素 (GnRH)及促性腺激素释放激素受体 (GnRH R)并细胞定位 ,试图从转录及翻译水平讨论人甲状腺可否合成GnRH及GnRH R ,为探讨GnRH可能影响甲状腺的功能提供形态学依据。　方法　采用免疫组织化学法和原位杂交技术。　结果　所测人正常甲状腺组织均呈较强的GnRH和GnRH R免疫反应阳性 ,甲状腺滤泡上皮细胞、滤泡旁细胞均为阳性细胞。免疫反应阳性物质主要分布在阳性细胞胞质内 ,胞核呈阴性。原位杂交结果同免疫组织化学染色基本一致 :甲状腺滤泡上皮细胞胞质和滤泡旁细胞胞质可检测到较强的GnRHmRNA及GnRH RmRNA阳性杂交信号。胞核均呈阴性 ,未检测到杂交信号。　结论　人甲状腺不仅表达GnRH和GnRH R ,而且可以自身合成GnRH和GnRH R。GnRH可能以自分泌的方式影响甲状腺的生理功能     

主要组织相容性复合体在人乳腺癌组织的表达

目的通过观察主要组织相容性复合体(MHC)在乳腺癌组织的表达,探讨MHC与乳腺癌发生发展的关系,为临床进行乳腺癌的生物治疗提供实验依据。方法收集手术切除的人乳腺癌组织和癌旁相对正常乳腺组织,用免疫组织化学方法和图像分析技术,观察人白细胞抗原(HLA-DR)阳性细胞、CD4+T细胞和TCD8+T细胞在不同乳腺组织中的表达。结果在乳腺癌组织中HLA-DR阳性细胞平均光密度值和面数密度明显高于正常乳腺组织(P<0.05);CD4+T和TCD8+T细胞阳性细胞面数密度与正常乳腺组织相比明显减少(P<0.05),且CD4+T细胞多分布在HLA-DR阳性细胞附近。结论人乳腺癌局部微环境中HLA-DR阳性细胞数多于正常乳腺组织,CD4+T和TCD8+T在人乳腺癌组织低于乳腺正常组织。     

不同生殖状态下雌性大鼠下丘脑视上核、室旁核内一氧化氮合酶活性的观察

为了进一步研究一氧化氮(NO)与生殖的关系,本文应用一氧化氮合酶(NOS)免疫组织化学ABC法和计算机图像分析,对正常、晚期(22天)妊娠和中期(7天)哺乳的大鼠下丘脑视上核、室旁核内NOS阳性神经元的细胞数、细胞平均切面面积及平均灰度值进行了观察。结果表明:晚期孕鼠的NOS阳性细胞数、细胞的平均切面面积和平均灰度值在室旁核分别为49.8±3.9、152.4±14.1μm2和153.4±8.9;在视上核分别为29.2±3.7、163.5±13.8μm2和140.5±7.2,前两项指标明显高于正常组(p<0.01),平均灰度值明显低于正常组(p<0.01)。中期哺乳鼠室旁核的NOS阳性细胞…     

甲状腺髓样癌的组织病理学和免疫组织化学的研究

为探讨甲状腺髓样癌的多分化，用免疫组织化学方法对２４例甲状腺髓样癌进行研究。结果显示：癌组织中见甲状腺球蛋白阳性细胞３例、降钙素阳性细胞２４例、铬粒素Ａ阳性细胞２３例、５－羟色胺阳性细胞１４例、神经元特异性烯醇化酶和生长抑素阳性细胞均为１０例、血管活性肠肽阳性细胞４例、Ｓ－１００蛋白阳性细胞１例。本组癌的多种抗原表达提示，它可能来源于可以产生多种激素的神经外胚叶细胞。另有３例见甲状腺球蛋白阳性癌细胞，提示：甲状腺髓样癌伴滤泡上皮分化和混合性髓样癌并不少见，可能来源于能向两个方向分化为滤泡上皮细胞和滤泡旁细胞的后腮干细胞。     

沙眼衣原体感染后大鼠卵巢S-100阳性树突状细胞的变化

目的　研究沙眼衣原体(CT)感染后,大鼠卵巢树突状细胞(DC)的变化。方法　成年Wistar雌 性大鼠,单侧卵巢囊接种沙眼衣原体E型株,于感染第1/2d、7d、14d取材,用免疫组化方法检测大鼠卵巢 DC的变化。结果　CT感染后大鼠卵巢黄体内S 100+阳性DC平均光密度值明显高于对照组(P<0.05或 0.01)。结论　沙眼衣原体感染可引起大鼠卵巢黄体内S 100阳性DC的表达。     

心理应激后大鼠腺垂体促甲状腺素细胞的定量观察

目的　探讨心理应激对腺垂体促甲状腺素细胞的影响。方法　采用心理应激模型 ,并在不同应激条件下对大鼠腺垂体促甲状腺素细胞进行免疫组织化学和电镜观察 ,并用体视学公式作定量分析。结果　应激大鼠促甲状腺素细胞体积密度 (Vv)和数密度 (Nv)的值均低于正常对照组 ,且在不同应激条件的A、B、C三组存在显著差别。结论　心理应激可不同程度地抑制大鼠腺垂体促甲状腺素细胞合成和分泌促甲状腺素。     

降钙素基因相关肽对局灶性脑缺血再灌注大鼠脑组织P53蛋白表达的调节作用

目的　研究降钙素基因相关肽(CGRP)对局灶性脑缺血再灌注大鼠脑组织P53蛋白表达的影 响,探讨降钙素基因相关肽对脑组织缺血再灌注损伤的保护作用及机制。方法　用线栓法制备大鼠大脑中动 脉阻塞(MCAO)模型,应用免疫组化和显微图像分析方法检测大鼠脑缺血再灌注后脑组织P53蛋白的表达。 结果　假手术组海马和顶叶内未见P53阳性细胞,脑缺血再灌注组阳性细胞明显增多,注射CGRP后P53阳 性细胞平均灰度值明显高于缺血再灌注组(P<0.01)。结论　CGRP下调缺血神经元P53蛋白的表达,可能 对缺血神经元的恢复有促进作用。     

心理应激对大鼠胃窦生长抑素mRNA细胞的影响

<正> 为探讨心理因素对胃窦粘膜生长抑素mRMA细胞的影响,本文采用Yanjin等创建的方法,以雄性SD大鼠15只,随机等分成 A、B、C三组,建立心理应激动物模型.以原位杂交组织化学技术和微机图像分析技术(Tiger图像分析软件),检测大鼠心理应激后胃窦粘膜生长抑素mRNA阳性细胞的表达强度和细胞体积密度(Vv)、单位体积数密度(Nv)的变化.结果表明:A、B两且大鼠胃窦粘膜生长抑素mRNA细胞的阳性表达信号明显强于只给无规律闪兴刺激的C组;阳性细胞的Vv和Nv方面,三组相比,为闪光间期长短不均加电刺激组(A组)>闪光间期衡定加电刺激组(B组)>单纯闪光间期长短不均组(C组).此结果揭示了心理应激后胃窦粘膜D细胞合成、分泌生长抑素是增加的.本实验从基因的量变角度揭示了心理应激后胃窦粘     

Immunohistochemical comparison of chromogranins A and B and secretogranin II with calcitonin and calcitonin gene-related peptide expression in normal, hyperplastic and neoplastic C-cells of the human thyroid

Normal and hyperplastic thyroid C-cells and 14 cases of medullary thyroid carcinoma were investigated immunohistochemically with antibodies against chromogranins A and B, secretogranin II, calcitonin and calcitonin gene-related peptide (CGRP). Normal and hyperplastic C-cells showed strong calcitonin and chromogranin A immunoreactivity whereas CGRP, chromogranin B and secretogranin II expression was less intense. Strong calcitonin and chromogranin A immunoreactivity was also found in the majority of tumour cells in medullary thyroid carcinoma. The CGRP, chromogranin B and secretogranin II staining observed was present in variable patterns. In some cases CGRP, chromogranin B and secretogranin II could only be demonstrated in isolated tumour cells with elongated processes suggestive of neuronal differentiation of these cells. The biological function(s) of the chromogranins/secretogranins remain(s) still unclear. There is evidence that these proteins are pro-peptides which give rise to functionally active compounds. Studies on normal C-cells and medullary thyroid carcinoma may elucidate the role of chromogranins/secretogranins in endocrine and neuronal cells.     

Somatostatin immunoreactive C cells in thyroid glands from various mammalia species

The relative distribution of somatostatin- and calcitonin-containing cells in thyroid glands from various mammalian species was investigated by immunoperoxidase staining, and the concentration of immunoreactive somatostatin by radioimmunoassay. In the thyroid glands of guinea pigs and rabbits, most of the calcitonin cells were also immunoreactive to the somatostatin antiserum, and high concentration of immunoreactive somatostatin was obtained. On the other hand, in the thyroids of other animal species—rats, dogs, pigs, cows, goats, cats, monkeys, mice, and hamsters—only a few C cells revealed the immunoreaction for somatostatin, and the concentration of somatostatin was low. In all animal species studied, the somatostatin was present in the same cells that contain calcitonin, though in guinea pigs and rats there were some C cells containing a large number of reaction products for somatostatin but very few for calcitonin. Thus, it was concluded that there was a considerable variation in somatostatin immunoreactivity of thyroid C cells from species to species.     

Effects of ovariectomy on parafollicular cells in the rat

Changes in the parafollicular cells in the thyroid gland of ovariectomized rats were investigated to clarify the relationship between the secretory function of sex hormones and that of parafollicular cells. Compared with control rats, the ovariectomized rats exhibited decreases in 1) the number of parafollicular cells in the thyroid glands, 2) the number of secretory granules in the parafollicular cells, and 3) the area occupied by the Golgi complex. These results suggest that the lack of estrogen caused by ovariectomy reduces the synthesis and release of calcitonin in parafollicular cells, which may be one of the causes of osteoporosis.     

C cells evolve at the same rhythm as follicular cells when thyroidal status changes in rats

C cells are primarily known for producing calcitonin, a hypocalcemic and hypophosphatemic hormone. Nevertheless, besides their role in calcium homeostasis, C cells may be involved in the intrathyroidal regulation of follicular cells, suggesting a possible interrelationship between the two endocrine populations. If this premise is true, massive changes induced by different agents in the activity of follicular cells may also affect calcitonin-producing cells. To investigate the behaviour of C cells in those circumstances, we have experimentally induced two opposite functional thyroid states. We hyperstimulated the follicular cells using a goitrogen (propylthiouracil), and we suppressed thyroid hormone synthesis by oral administration of thyroxine. In both scenarios, we measured T₄, TSH, calcitonin, and calcium serum levels. We also completely sectioned the thyroid gland, specifically immunostained the C cells, and rigorously quantified this endocrine population. In hypothyroid rats, not only follicular cells but also C cells displayed hyperplastic and hypertrophic changes as well as increased calcitonin levels. When exogenous thyroxine was administered to the rats, the opposite effect was noted as a decrease in the number and size of C cells, as well as decreased calcitonin levels. Additionally, we noted that the two cell types maintain the same numerical relation (10 ± 2.5 follicular cells per C cell), independent of the functional activity of the thyroid gland. Considering that TSH serum levels are increased in hypothyroid rats and decreased in thyroxine-treated rats, we discuss the potential involvement of thyrotropin in the observed results.     

The physiological role of calcitonin assessed through chronic calcitonin deficiency in rats

1. An attempt was made to assess the physiological role of calcitonin by producing its prolonged deficiency in rats.2. Three groups of young rats viz. (I) sham-operated controls, (II) thyroidectomized with thyroxine replacement and (III) thyroidectomized with thyroxine as well as calcitonin replacement, all with parathyroid autografts, were studied for 5 weeks. Groups II and III showed impaired Ca tolerance and group II also showed significant reduction in total (organic and inorganic) mass of femora as compared to group I.3. Three similar groups of old mature rats were studied as under (2) for 9 weeks. Groups II and III in this case showed no significant changes as compared to group I.4. In intact mature rats, Ca tolerance is impaired, thyroid content of calcitonin is high and the hypocalcaemic action of calcitonin is much reduced as compared to intact young rats.5. It is concluded that calcitonin plays a useful but non-essential role in Ca metabolism and bone development of young growing rats. Its function, if any, in mature rats on a normal diet remains doubtful.     

Calcitonin gene-related peptide gene expression in collagen-induced arthritis is differentially regulated in primary afferents and motoneurons: influence of glucocorticoids.

Calcitonin gene-related peptide is involved in peripheral and spinal mechanisms of inflammatory pain. In this paper, we used collagen II-induced arthritis in the rat as a model to investigate the influence of chronic arthritic pain on calcitonin gene-related peptide gene expression in sensory and motor pathways. Additionally, we examined the effect of the glucocorticoid drug budesonide on arthritis-induced changes of calcitonin gene-related peptide expression and constitutive calcitonin gene-related peptide expression. Thirteen days after the immunization with native rat collagen type II rats developed a progressive and chronic polyarthritis which was scored with respect to the degree of swelling and/or redness of the paw and ankle joints. Budesonide significantly attenuated the extent of arthritis. Changes in calcitonin gene-related peptide expression were evaluated by semiquantitative in situ hybridization and immunocytochemistry on day 21 post-immunization. In sensory neurons of dorsal root ganglia of arthritic rats, a significant increase in calcitonin gene-related peptide messenger RNA and protein levels was seen. These increases were completely blocked by budesonide. Also in dorsal root ganglia of non-arthritic rats, budesonide had an effect, with reduced calcitonin gene-related peptide messenger RNA levels below constitutive concentrations. Image analysis of calcitonin gene-related peptide immunoreactivity revealed that changes in calcitonin gene-related peptide expression were due to alterations in calcitonin gene-related peptide expression levels rather than to de novo synthesis or changes in the numbers of calcitonin gene-related peptide expressing neurons. In spinal motoneurons of arthritic rats, marked decreases in calcitonin gene-related peptide messenger RNA and protein levels were measured. These reductions were attenuated by budesonide. The changes in calcitonin gene-related peptide expression in motoneurons correlated with the severity of arthritis in the ipsilateral hind paw. Budesonide had no effects on calcitonin gene-related peptide messenger RNA levels in motoneurons of non-arthritic rats. The opposite regulation of calcitonin gene-related peptide gene expression in primary sensory and spinal somatomotor pathways in collagen-induced arthritis suggests that calcitonin gene-related peptide plays a specific role in both chronic inflammatory pain and arthritis-induced motor dysfunction. The sensitivity of constitutive and inflammation-induced sensory calcitonin gene-related peptide expression to budesonide treatment may indicate that the beneficial effects of steroid treatment in inflammation is partly mediated by down-regulation of calcitonin gene-related peptide in sensory neurons involved in neurogenic inflammation.     

Comparative study of changes in activity of calcitonin-producing cells in the thyroid and lungs during hypercalcemia

The conflicting results of previous evaluations of the influence of hypercalcemia on thyroid C-cells and the absence of reports on its influence on calcitonin (CT)-producing cells of the airways prompted this study. It aims to evaluate the activity and changing dynamics in CT-producing cells in rat thyroid glands and lungs after prolonged hypercalcemia. The hypercalcemia was induced by an intraperitoneal injection of 100,000 IU of vitamin D₃ (Vigantol^®, Merck). Control and experimental rats were examined after 1, 7 and 14 days. Calcium and CT concentrations were measured in blood plasma. Paraffin wax embedded sections of thyroid and lungs were stained with hematoxylin and eosin. CT was detected immunohistochemically in thyroid C-cells and lung endocrine cells. Results showed increased calcium and CT concentrations in rat blood after vitamin D₃ administration, a statistically significant reduction in number of CT-positive cells in the thyroid with indications of their increased activity, and a statistically non-significant reduction of the number of cells immunopositive for CT in lungs after 1 day, and an increase after 7 days, compared to the control group. Lack of an obvious correlation in lung neuroendocrine cells among the rats with hypercalcemia may indicate the presence of other, perhaps local, functions for calcitonin.     

Somatostatin-immunoreactive cells in medullary carcinoma of the thyroid.

Ten medullary carcinomas of the thyroid were examined by immunohistochemistry using antisera against calcitonin, somatostatin, and ACTH. In addition to calcitonin cells, eight of the tumors contained somatostatin-immunoreactive cells in varying numbers. No ACTH-immunoreactive cells were found. The results indicate that thyroid medullary carcinomas often are multihormonal, and that somatostatin is common in such tumors. In contrast to calcitonin cells, somatostatin cells are not found in normal human thyroid tissue.