IL—10基因在人脑胶质瘤中的表达及意义

采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）方法检测了白细胞介素－１０（ＩＬ－１０）ｍＲＮＡ在人脑胶质瘤细胞中的表达。结果１７例患者的新鲜脑胶质瘤标本中，有１５例表达ＩＬ－１０ｍＲＮＡ，提示免疫抑制因子ＩＬ－１０可能是导致脑胶质瘤患者免疫功能低下的原因之一。     

基质金属蛋白酶-2 mRNA在胃癌中的表达

目的　探讨基质金属蛋白酶２（ＭＭＰ２） ｍＲＮＡ 及蛋白在胃癌中的表达。方法　用ＲＴＰＣＲ 法及免疫组化法分别检测２５ 例胃癌患者癌组织及癌旁２ｃｍ 与≥５ｃｍ 组织ＭＭＰ２ ｍＲＮＡ 及蛋白的表达。结果　胃癌组织ＭＭＰ２ ｍＲＮＡ 表达率（１７／２５） 高于两组癌旁组织，与癌旁≥５ｃｍ 组织表达率（９／２５）比较差异有显著性（Ｐ＜０ ．０５）；免疫组化显示胃癌组织ＭＭＰ２ 蛋白的表达率（１７／２５） 也高于两组癌旁组织，与癌旁≥５ｃｍ 组织表达率（８／２５） 比较差异有显著性（Ｐ＜ ０ ．０５） 。结果接近ＲＴＰＣＲ 法；胃癌组织ＭＭＰ２ ｍＲＮＡ 表达值显著高于两组癌旁组织（Ｐ均＜０ ．０１）；ＭＭＰ２ ｍＲＮＡ 及蛋白表达似与胃癌分化及临床分期有关。１９ 例进展期胃癌组织及癌旁２ｃｍ 组织，ＭＭＰ２ ｍＲＮＡ 表达值显著高于６ 例早期胃癌相应组织（Ｐ均＜ ０．０５） 。结论　ＭＭＰ２ 在胃癌侵袭及转移过程中可能发挥重要作用。     

苯那普利对糖尿病肾小球细胞外基质增生的抑制作用？…

目的：了解血管转换酶抑制苯那普利对单侧肾功能除糖尿病大鼠肾脏结构和功能改变的保护作用及机制。方法：应用生物化学分析，ＲＴ－ＰＣＲ方法检测苯地普利治疗对糖尿病大鼠血尿素氮，肌肝，血脂，肾小球细胞外基质（ＥＣＭ）以及肾皮质内金属蛋白酶－３（ＭＭＰ－３）的ｍＲＮＡ表达的影响，结果：糖尿病大鼠的尿素氮，肌酐及甘油三酯，胆固醇水平均较非糖尿病大鼠明显增加，ＰＡＳ染色发现肾小球系膜区ＥＣＭ明显积聚，苯那普利治     

糖皮质激素对哮喘患者白细胞介素—5mRNA表达与嗜酸性粒细胞激活作…

为了观察糖皮质激素对哮喘患者白细胞介素（ＩＬ）－５ｍＲＮＡ表达与嗜酸性粒细胞激活作用的影响，本研究用逆转录（ＲＴ）－聚合酶链反应（ＰＣＲ）法半定量分析了哮喘患者用糖皮质激素治疗前后外周血单个核细胞（ＰＢＭＣ）中ＩＬ－５ｍＲＮＡ表达水平的变化，检测了血清嗜酸细胞阳离子蛋白（ＥＣＰ）浓度、一秒钟用力呼气容积（ＥＦＶ１）下降２０％时的乙酰甲胆碱（ＭＣＨ）激发浓度（ＭＣＨ－ＰＣ２０值）和基础ＦＥＶ１占用力     

原发性肾病综合征患者IL—3血浆水平和基因表达的研究

目的：探讨成人原发性肾病综合征（ＮＳ）患者外周血单个核细胞（ＰＢＭＣ）中ＩＬ－１３ｍＲＮＡ表达及ＩＬ－１３血浆水平变化。方法：选择１０名健康正常对照者和１６名ＮＳ患者。采用反转录多聚酶链反应（ＲＴ－ＰＣＲ）技术，对ＮＳ患者ＰＢＭＣ中ＩＬ－１３ｍＲＮＡ表达量进行分析。同时应用ＩＬ－１３特异的酶联免疫吸附法（ＥＬＩＳＡ）测定ＩＬ－１３血浆水平。结果：ＮＳ患者ＰＢＭＣ中ＩＬ－１３ｍＲＮＡ表达量及血浆ＩＬ     

一氧化氮与门脉高压性肠病关系的实验研究

目的探讨ＮＯ在门脉高压性肠病发病机理中的作用。方法制作大鼠四氯化碳门脉高压模型。第１２周末，在冰冻切片上进行结肠ＮＡＤＰＨ黄递酶组织化学染色；应用化学发光法测定ＮＯＳ活性；ＲＴ－ＰＣＲ反应测定ＮＯＳｍＲＮＡ表达量。结果（１）ＮＡＤＰＨ黄递酶组化染色：模型组大鼠结肠粘膜下小血管内皮细胞和神经纤维的ＮＯＳ染色强度显著高于正常对照组。而表层上皮细胞ＮＯＳ染色减弱。（２）化学发光法测定ＮＯＳ活性：模型组大鼠结肠粘膜ＮＯＳ活性较正常对照组显著增加，其中ｉＮＯＳ活性增加更为明显。（３）ＲＴ－ＰＣＲ：模型组大鼠结肠粘膜ＮＯ－ＳｍＲＮＡ表达量较正常对照组显著增加。结论ＮＯ可能由于其扩张血管的特性，而参与门脉高压性肠病血管病变的发病过程，并与门脉高压性肠病粘膜病变、可能存在的运动功能异常亦有一定的关系。     

SHR血管平滑肌PDGF—A链,TGFβ1及其受体mRNA表达的研究

目的：在自发型高血压和正常血压的Ｖｉｓｔａｒ－ｋｙｏｔｏ大鼠血管壁平滑肌细胞（ＳＨＲ－ＶＳＭＣ和ＷＫＹ－ＶＳＭＣ）,比较几种内源性生长因子及它们受体的表达水平,从ＶＳＭＣ自分泌和旁分泌产生骨源性生长因子的角度,阐述ＳＨＲ－ＶＳＭＣ和ＷＫＹ－ＶＳＭＣ的差异。方法：以定量ＲＴ－ＰＣＲ技术,在二株系ＶＳＭＣ,检测长型ＰＤＧＦ－Ａ ,ＴＧＦβ１及它的受体ｍＲＮＡ的表达水平。结果：（１）在ＳＨＲ－ＶＳＭＣ,     

辛伐他汀对大鼠肾间质成纤维细胞的影响

目的：研究３－羟基－３－甲基戊二酰辅酶Ａ（ＨＭＧ－ＣｏＡ）还原酶抑制剂（辛伐他汀）对大鼠肾间质成纤维细胞（ＲＦＢ）增生，细胞周期及细胞间信号传导的影响。方法：应用ＭＴＴ比色法、流式细胞仪、半定量ＲＴ－ＰＣＲ等技术，分别检测不同浓度辛伐他汀对体外培养的ＲＦＢ的增生、细胞周期以及细胞信号传导中ｃ－ｆｏｓ ｍＲＮＡ表达的影响。结果：辛伐他汀可抑制ＲＦＢ的增生活性（Ａ值），尤以无血清组显著，各浓度加药组Ａ组与正常对照组比较差异显著（Ｐ〈０．０５）；各浓度加药组Ｇ０／Ｇ１期细胞比增高，Ｓ期细胞比、ＰＩ值和ＤＮＡ含量则逐渐下降，Ｇ２／Ｍ期细胞比变化不明显；各浓度加药组ＲＦＢ ｃ－ｆｏｓ ｍＲＮＡ的表达逐渐下降，并呈一定的剂量依赖性，与正常对照组比较差异显著（Ｐ〈０．０５）。结论：辛伐他汀可抑制细胞增生，阻止ＲＦＢ由ＧＩ期进     

支气管哮喘患者粒—基噬细胞集落刺激因子mRNA表达的研究

目的 探讨利用无创伤性方法寻找较为特异性地反映哮喘患气道炎症的客观指标。方法 通过研究哮喘患气道粘膜细胞粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）ｍＲＮＡ表达,并以此为参照,分析哮喘患诱导痰细胞ＧＭ－ＣＳＦ ｍＲＮＡ变化,利用免疫组化结合逆转录聚合酶链反应（ＲＴ－ＰＣＲ）技术,分析哮喘组（１０例）气道粘膜细胞ＧＭ－ＣＳＦ ｍＲＮＡ的表达;用３％高渗盐水诱导痰液,利用ＲＴ－ＰＣＲ技术分析哮喘组（     

非小细胞肺癌淋巴转移与nm23—H1基因突变及mRNA表达异…

目的 观察非小细胞肺癌（ＮＳＣＬＣ）组织中ｎｍ２３－Ｈ１基因的存在状况及其转录表达水平，分析肺癌恶性转移与该基因异常的关系。方法 采用ＰＣＲ－ＳＳＣＰ和半定量ＲＴ－ＰＣＲ方法，以正常癌旁组织及正常肺组织为对照，观察了３１例原发性ＮＳＣＬＣ中ｎｍ２３－Ｈ１基因的突变和ｍＲＮＡ表达情况。结果 ３１例肺癌中未发现１例存在ｎｍ２３－Ｈ１基因突变。在２０例具淋巴结转移的ＮＳＣＬＣ中ｎｍ２３－Ｈ１基因ｍＲＮＡ     

Phenylethanolamine N-methyltransferase-containing neurons in the limbic system of the young rat.

Fifteen years ago epinephrine cells were shown to be present in the medulla oblongata of the rat. These cell groups (C1 and C2) were thought to supply the epinephrine innervation in the rest of the central nervous system. In this study I demonstrate the presence of epinephrine-producing neurons in the forebrain of the young rat. Neurons that are immunopositive for phenylethanolamine N-methyltransferase (S-adenosyl-L-methionine:phenylethanolamine N-methyltransferase, EC 2.1.1.29) are present in the central nucleus of the amygdala as well as in the bed nucleus of the stria terminalis. Neurons in the same location are also immunopositive for tyrosine hydroxylase [tyrosine 3-monooxygenase; L-tyrosine, tetrahydrobiopterine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2]. The phenylethanolamine N-methyltransferase immunopositivity disappears by day 35, while a small amount of tyrosine hydroxylase-positive cells still can be found in the adult. In situ hybridization reveals tyrosine hydroxylase mRNA in the above nuclei in both young and adult animals. The number of the positive cells decreases in adulthood. RNA blot-hybridization analysis showed the presence of phenylethanolamine N-methyltransferase mRNA in the amygdala and the bed nucleus of the stria terminalis in the young and in the adult rat brain. Neurons that are immunopositive for phenylethanolamine N-methyltransferase are also present in the human amygdala.     

Molecular characterization of an aquaporin cDNA from brain: candidate osmoreceptor and regulator of water balance.

The aquaporins transport water through membranes of numerous tissues, but the molecular mechanisms for sensing changes in extracellular osmolality and regulating water balance in brain are unknown. We have isolated a brain aquaporin by homology cloning. Like aquaporin 1 (AQP1, also known as CHIP, channel-forming integral membrane protein of 28 kDa), the deduced polypeptide has six putative transmembrane domains but lacks cysteines at the known mercury-sensitive sites. Two initiation sites were identified encoding polypeptides of 301 and 323 amino acids; expression of each in Xenopus oocytes conferred a 20-fold increase in osmotic water permeability not blocked by 1 mM HgCl2, even after substitution of cysteine at the predicted mercury-sensitive site. Northern analysis and RNase protection demonstrated the mRNA to be abundant in mature rat brain but only weakly detectable in eye, kidney, intestine, and lung. In situ hybridization of brain localized the mRNA to ependymal cells lining the aqueduct, glial cells forming the edge of the cerebral cortex and brainstem, vasopressin-secretory neurons in supraoptic and paraventricular nuclei of hypothalamus, and Purkinje cells of cerebellum. Its distinctive expression pattern implicates this fourth mammalian member of the aquaporin water channel family (designated gene symbol, AQP4) as the osmoreceptor which regulates body water balance and mediates water flow within the central nervous system.     

Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland

Distribution and development of growth hormone secretagogue receptor (GHS-R) mRNA expression in rat brain and pituitary gland were examined using ribonuclease protection assay. In adult male rats, GHS-R mRNA levels were highest in the pituitary gland, whereas those in the hypothalamus and hippocampus were 57 and 30% of those in the pituitary gland, respectively. Less abundant but detectable levels of GHS-R mRNA were found in the midbrain, pons, and medulla oblongata, but expression was barely detectable in the cerebellum and cerebral cortex. The expression of GHS-R mRNA was detected at late gestation (embryonic day 19) in the pituitary gland, hypothalamus, and brainstem. The mRNA levels increased with age in the pituitary gland, and decreased postnatally in the brainstem, while they remained constant in the hypothalamus during development. In contrast, GHS-R mRNA was not detectable in the hippocampus during the fetal period, but was first detected on postnatal day 7. Expression of GHS-R mRNA was also examined in the spontaneous dwarf rat (SDR), a model for isolated GH deficiency, to examine alterations in GHS-R mRNA expression in a GH-deficient state. GHS-R mRNA levels in the pituitary gland of SDRs were higher than those of control rats, suggesting negative regulation of GHS-R mRNA by GH in this region. GHS-R mRNA levels increased in the hypothalamus of female, but not in male SDRs. In contrast, GHS-R mRNA levels were not affected by GH in the brainstem and hippocampus. These results indicate that region-specific, developmentally regulated expression of GHS-R mRNA may reflect divergent physiological roles of GHS/GHS-R in distinct regions of the central nervous system and the pituitary gland.     

Oxyntomodulin and glicentin: brain-gut peptides in the rat

Glucagon-like materials and glucagon have been identified by immunoassay and immunocytochemistry in the mammalian central nervous system. However, the molecular forms relevant to brain glucagon-like immunoreactivity (GLI) have not been precisely defined. In the rat small intestine, more than 90% of GLI is constituted by two peptides: oxyntomodulin (OXM) and glicentin. This work was initiated to characterize and determine the concentrations of these two peptides and glucagon in the rat central nervous system and to compare their relative proportions with those found in the gut. Different regions from the adult rat brain were analyzed by HPLC in association with RIA, using a central glucagon antiserum and an antibody directed toward the C-terminal end of OXM and glicentin. The elution profiles of hypothalamus extracts were constituted by two main peaks, both detected by the two antibodies used and displaying the same retention times as glicentin and OXM, respectively. A third small peak, which coeluted with glucagon, was constantly recorded with the central glucagon antiserum. The percentages of glicentin, OXM, and glucagon in 10 hypothalami were 37 +/- 1%, 55 +/- 1%, and 8 +/- 2%, respectively (n = 8). This distribution was quite similar to that in small intestinal extracts (38 +/- 1%, 59 +/- 1%, and 1.3 +/- 0.1%, respectively; n = 7); however, the peptide concentrations were almost 50-fold greater in intestine than in hypothalamus. In the medulla oblongata, the same peptide ratio was observed, with 10-fold lower concentrations compared to those in hypothalamus. In olfactory bulb, cerebellum, and cortex the concentrations were close the the detection limit, whereas they could be not detected in the pituitary. The combination of HPLC and specific RIAs allowed us to unambiguously characterize OXM and glicentin as the major components of GLI in the rat hypothalamus and medulla oblongata. The same proportion of these two peptides in the central nervous system and the gut indicates that a similar posttranslational processing exists in these rat tissues, another example of the brain-gut axis.     

Molecular cloning and expression of equine calcitonin,calcitonin gene-related peptide-I,and calcitonin gene-related peptide-II

In this study, we describe the cloning and tissue expression of equine calcitonin (CT), calcitonin-gene related peptide (CGRP)-I, and CGRP-II cDNA. We also describe a novel divergent form of CGRP (CGRP-I). Equine CT has greatest homology (>85%) to human, rat and mouse subgroups of calcitonins. Equine CGRP-I has low homology (<59%) to CGRPs of other species. The signal and N-terminal peptides for equine CT and CGRP-I were identical, indicating that these peptides are encoded by a gene equivalent to the human CALC-I gene. Equine CGRP-II has >80% homology to chicken, human, rat, ovine, swine, and bovine CGRPs. The homology between equine CGRP-I and CGRP-II is low (56%). The high homology of equine CGRP-II and the low homology of equine CGRP-I to CGRP in other species were unexpected findings. Northern blot analysis revealed that CT mRNA expression was restricted to the thyroid gland; however, RT-PCR revealed that CT mRNA expression was also present in the pituitary gland and in the liver. CGRP-I and CGRP-II mRNA expression was present in several regions of the nervous system and other tissues of neuroectodermal origin. An unexpected finding was CGRP-I expression in the kidney by both Northern analysis and by RT-PCR. Based on these results, CT gene expression in the horse was not restricted to the thyroid gland, and CT may be important in regulating pituitary cell function. CGRPs are widely expressed in tissues of the central and peripheral nervous system. Information from this study will be valuable to study the role of CT, CGRP-I, and CGRP-II in equine health and disease.     

Prehepatic portal hypertension in rats modifies norepinephrine metabolism in hypothalamus medulla oblongata and portal vein

The present experiments investigated the possible relationship between portal hypertension and norepinephrine metabolism in the central nervous system (hypothalamus and medulla oblongata) and the portal vein in the rat. Group I (72), portal hypertensive, and group II (70) sham-operated animals, were sacrificed day 14, and endogenous norepinephrine content, uptake and release from hypothalamus, medulla oblongata, and portal vein were investigated. In group I our results showed increases in norepinephrine storage (69%; 8.3%) and release (19.7%; 43.8%) and a diminished uptake (42.3%; 27.5%) in the hypothalamus and medulla oblongata, respectively. Portal veins showed a decreased content and uptake (62.5% and 43.5%, respectively) and increased release (25%) compared to group II rats. These results suggest a close relationship between the central nervous system and rat portal hypertension, perhaps related to modifications of central sympathetic activity.This work was supported in part by Consejo de Investigaciones Técnicas y Cientificas of Argentina (CONICET) by grants PIA 2042/90 and 0738/91-010 and by the University of Buenos Aires grant FA 031.