风湿性心脏病心力衰竭患者抗心脏β_1和M_2受体自身抗体的研究

目的：探讨抗心脏Ｇ－蛋白偶联型β１ 和Ｍ２ 受体的自身抗体是否与风湿性心脏病（ＲＶＨＤ）心力衰竭有关。方法：以心脏Ｇ－蛋白偶联受体β１ 和Ｍ２ 细胞外第二环表位肽段序列的合成肽作为抗原,应用酶联免疫吸附测定（ＥＬＩＳＡ）技术检测５０ 例ＲＶＨＤ患者和４１ 例正常人血清中抗心脏β１ 和Ｍ２ 受体的自身抗体。结果：５０ 例ＲＶＨＤ心力衰竭患者中有２０ 例存在抗心脏β１ 受体自身抗体（４０．０％ ）,正常组仅５ 例（１２．２％ ）;前组又有２４ 例存在抗心脏Ｍ２ 受体自身抗体（４８．０％ ）,正常组仅４ 例（９．８％ ）。ＲＶＨＤ组β１ 和Ｍ２ 自身抗体的滴度分别为１∶１０４ 与１∶１１７,正常组为１∶４０ 与１∶２４（Ｐ＜ ０．０５）;心功能Ⅱ～Ⅲ级的阳性率及抗体滴度也明显高于心功能Ⅳ级。结论：抗心脏β１ 和Ｍ２ 受体的自身抗体与ＲＶＨＤ有关,这两种自身抗体可能参与ＲＶＨＤ心力衰竭的病理改变。     

高血压性心脏病抗G-蛋白偶联β_1-肾上腺素能受体与M_2-胆碱能受体自身抗体的初步研究

目的抗心肌β１和Ｍ２的自身抗体是否与高血压性心脏病有关。方法以心脏Ｇ－蛋白偶联受体β１和Ｍ２细胞外第二环表位肽段的合成肽作为抗原，应用酶联免疫吸附测定（ＥＬＩＳＡ）技术检测４４例高血压性心脏病（ＨＣＤ）患者、３６例单纯性高血压患者和４１例正常人（对照组）血清中抗β１和Ｍ２受体的自身抗体。结果抗心肌β１受体自身抗体的检出率，高血压性心脏病患者为４７．７％（２２／４４），显著高于高血压患者１１．１％（４／３６）与正常人１２．１（５／４１）；抗心肌Ｍ２受体自身抗体的检出率，高血压性心脏病患者为４５．５％（２０／４４），显著高于高血压患者８．３％（３／３６）与正常人９．８％（４／３６）。高血压性心脏病患者血清中抗β１和Ｍ２受体的自身抗体滴度分别为１∶９６与１∶１０３，远高于单纯性高血压和对照组的１∶４０与１∶２４的相应抗体滴度（Ｐ＜０．０５）。结论高血压性心脏病患者不仅存在抗心肌β１和Ｍ２受体的自身抗体而且其抗体滴度也明显高于单纯性高血压及正常人，检测该抗体对预测原发性高血压患者是否并发心肌病变，可能有一定的参考价值。     

病毒性心肌炎与扩张型心肌病血清抗M受体抗体研究

目的Ｍ受体和肾上腺素能受体通过自主神经调节心脏功能。在某些病理条件下，血清抗Ｍ受体抗体的产生与疾病的发生、发展密切相关。本研究拟通过对病毒性心肌炎（ＶＭＣ）与扩张型心肌病（ＤＣＭ）患者血清抗Ｍ受体抗体的检测，探讨该指标的辅助判断价值。方法随机选择ＶＭＣ患者３２例、ＤＣＭ患者１２例、冠心病患者２０例及正常对照组１４例，受试者血清与人心室肌组织片预孵育后，冷冻切片法测定Ｍ受体与配基［３Ｈ］ＱＮＢ结合的Ｂｍａｘ与Ｋｄ值。ＶＭＣ患者血清同时进行心肌肌钙蛋白Ⅰ（ｃＴｎⅠ）测定与淋巴细胞肠道病毒逆转录扩增。结果７例正常人（意外死亡者）心室肌Ｍ受体与配基［３Ｈ］ＱＮＢ结合的Ｂｍａｘ值为５１．４±６．０ｆｍｏｌ／Ｌｍｇｐｒ，Ｋｄ值为０．６８±０．０９ｎｍｏｌ／Ｌ。ＤＣＭ患者血清孵育后Ｍ受体和配基结合的Ｂｍａｘ值下降（Ｐ＜０．０１），Ｋｄ值升高（Ｐ＜０．０１）；ＶＭＣ患者仅Ｋｄ值升高（Ｐ＜０．０１），但ｃＴｎＩ阳性及病毒扩增阳性者Ｂｍａｘ值同时下降（Ｐ＜０．０５）。结论ＶＭＣ和ＤＣＭ患者血清中存在抗Ｍ受体抗体，其导致心肌Ｍ受体和配基结合的数目与亲和力改变，对ＤＣＭ诊断以及ＶＭＣ向ＤＣＭ演变可能提供有益的证据。     

高血压性心脏病抗G—蛋白偶联β1—肾上朱素能受体与M2—胆碱能受体自身抗体

目的 抗体肌β１和Ｍ２的自身全是否与高血压怀心脏病有关。方法 以心脏Ｇ－蛋白偶联受体β１和Ｍ２细胞外第二环表位肽段的合成肽作为抗原，应用酶联免疫吸附测定（ＥＬＩＳＡ）技术检测４４例高血压性心脏病（ＨＣＤ）患者、３６例单纯性高血压患者和４１例正常（对照组）血清中抗β１和Ｍ２受体的自身抗体。结果 抗心肌β１受体自身抗体的检出率，高血压性心脏病患者为４７．７％（２２／２４），显著高于高血压患者１１．１％     

免疫转印法检测扩张型心肌病患者抗β－受体抗体的探讨

应用免疫转印法检测２０例扩张型心肌病（ＤＣＭ）患者血清中抗心肌β－肾上腺素能受体抗体。结果：抗心肌细胞膜６７ｋＤ肽（β－受体）抗体在１１例ＤＣＭ患者血清中被检出，而对照组均为阴性（Ｐ＜０．０１），检测的特异性１００％，敏感性５５％。提示免疫转印法检测抗心肌β－受体抗体，对扩张型心肌病具有临床诊断价值。     

高血压不同时期β受体的变化及其意义

目的探讨高血压病（ＥＨ）患者心肌β受体的变化与其病情的关系。方法采用３Ｈ－双氢阿普洛尔（３Ｈ－ｄｉｈｙｄｒｏａｌｐｒｅｎｏｌｏｌ）放射性配基结合分析法，测定６９例ＥＨ不同时期男性患者外周血淋巴细胞的β受体密度（与心肌β受体密度呈显著性正相关），并对各期１０例患者进行羟甲叔丁肾上腺素药物实验，以测定β受体敏感性。结果Ⅰ、Ⅱ期ＥＨ患者β受体的最大结合容量（βＭｍａｘ）分别较正常人增加３０．８％（Ｐ＜０．０１）和５６．７％（Ｐ＜０．００１），羟甲叔丁肾上腺素增加心率３０次／分的变时剂量（ＣＤ３０）分别较正常人降低２０．７％（Ｐ＜０．０１）和３７．９％（Ｐ＜０．００１），其中并发左室肥厚患者上述特征的显著性明显大于元左室肥厚患者（均Ｐ＜０．００１）。Ⅲ期ＥＨ患者βｍａｘ较正常人降低２７．２％，ＣＤ３０则较正常人增加２４．１％（均Ｐ＜０．０１），其中井发心力衰竭患者上述特征的显著性明显大于无衰竭患者（均Ｐ＜０．００１）。结论心肌β受体密度及功能的变化可能与ＥＨ及其并发左室肥厚与心力衰竭有关，提示，该参数可作为判断ＥＨ患者病情变化的一项指标。     

抗心肌线粒体自身抗体在扩张型心肌病诊断中的价值

采用ＤＩ－ＰＡＰ法和ＩＢＴ法，对心功能改变相似的２２例扩张型心肌病（ＤＣＭ）和２０例冠心病患者进行自身抗体检测，仅在ＤＣＭ患者血清中检测出抗心肌线粒体抗体（１５例阳性）和线粒体ＡＤＰ／ＡＴＰ载体抗体（１６例阳性），两种方法的符合率８６．４％，总计１７例阳性；而冠心病组和正常人对照组均为阴性（Ｐ＜０．０１）．提示抗心肌线粒体抗体的检测，对ＤＣＭ诊断有很高的特异性和敏感性，可作为ＤＣＭ诊断的重要指标．     

大肠癌患者外周血CD44表达的变化

目的探讨大肠癌患者外周血ＣＤ４４表达变化的意义．方法以流式细胞仪对２６例大肠癌患者外周血ＣＤ４４表达进行免疫荧光检测，通过自身对照分析术前、术后变化，并与正常人（ｎ＝２０）及大肠良性肿瘤患者（ｎ＝１７）进行对比研究，ｔ检验处理数据．结果大肠癌患者外周血ＣＤ４４细胞（２０５％±４５％）明显高于正常人（１８％±１１％）及大肠良性肿瘤患者（２４％±１４％，Ｐ＜００１），后二者差别不明显（Ｐ＞００５），已发生转移的大肠癌患者外周血ＣＤ４４含量（２２９％±４８％）较无转移者（１６１％±３３％）明显增高（Ｐ＜００１），无转移组根治手术后外周血ＣＤ４４含量迅速下降（２９％±１８％）至正常人水平，已有转移者变化不明显（Ｐ＞００５）．结论ＣＤ４４水平可作为大肠恶性肿瘤负荷及转移的指标．     

病毒性心肌炎可溶性白细胞介素2受体、自然杀伤细胞活性及T细胞亚群的检测

应用单克隆与多克隆双抗体夹心法检测３６例病毒性心肌炎（ＶＭＣ）及２４例正常人（ＮＣ）的血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ），同时测定外周血自然杀伤细胞（ＮＫＣ）活性和Ｔ淋巴细胞亚群。结果显示ＶＭＣ患者ｓＩＬ－２Ｒ明显高于ＮＣ组（Ｐ＜０．００１）．而ＮＫＣ活性明显低于ＮＣ组（Ｐ＜０．０１），Ｔ细胞亚群与ＮＣ组比较，急性ＶＭＣ患者总Ｔ细胞（ＣＤ３），辅助性Ｔ细胞（ＣＤ４）和抑制性Ｔ细胞（ＣＤ８）均减少，ＣＤ４／ＣＤ８比值显著降低（Ｐ＜０．０５．０．０１），以细胞免疫功能低下为明显；而ＶＭＣ后遗症期患者ＣＤ３、ＣＤ４与ＮＣ组无差异（Ｐ＞０．０５），ＣＤ８显著降低（Ｐ＜０．０５），ＣＤ４／ＣＤ８比值显著高于ＮＣ组（Ｐ＜０．０５），以细胞免疫调节失衡为主。上述结果提示细胞免疫功能低下及免疫功能失调为ＶＭＣ发病及影响预后的重要因素。     

扩张型心肌病白细胞介素2受体的初步研究

检测２０例扩张型心肌病（ＤＣＭ）及２０例正常人（ＮＣ）的血清可溶性白细胞介素２受体（ＳＩＬ－２Ｒ）及外周血单个核细胞膜白细胞介素２受体（ＩＬ－２Ｒ）的表达。结果发现，ＤＣＭ患者ＳＩＬ－２Ｒ明显高于ＮＣ组（Ｐ＜０．００１），而膜ＩＬ－２Ｒ表达低于ＮＣ组（Ｐ＜０．０１），提示ＩＬ－２Ｒ在ＤＣＭ发病中可能有一定意义。     

Prolactin receptors in primary cultures of carcinogen-induced rat-mammary tumors.

7,12-Dimethylbenz[alpha]anthracene-induced rat mammary tumors were dissociated with collagenase and hyaluronidase and placed into primary culture. In most cultures, specific binding of 125I-labeled ovine prolactin was (i) lower than that for the original tumors unless bovine prolactin (1 microgram/ml) had been added to the dissociation medium, and (ii) varied with the type of growth medium used. The level of prolactin binding in cultured cells was relatively constant for the first 7-10 days. Prolactin binding in cultured cell homogenates was maximal at pH 7.0, proportional to cell protein, specific for prolactin, and reached a steady state by 12 h at 22 degrees C. The half-maximum inhibition of 125I-labeled prolactin binding by unlabeled prolactin was 100 ng/ml for cells grown in 5-1000 ng of prolactin/ml. After prolactin was removed from the growth medium, the level of available binding sites progressively increased, reached a maximum at 48 h and then declined. At 48 h, the dissociation constant for prolactin binding (Kd approximately 1 x 10(-10) M) was comparable to that in tumors. In some cultured tumors, a 48-h treatment with 0.5 or 1.0 ng of prolactin/ml caused an apparent increase in the level of prolactin binding. Prolactin increased DNA synthesis and its removal caused a reduction in [3H]estradiol and [3H]-R5020 binding to cultured cell cytosols.     

Membrane depolarization causes a direct activation of G protein-coupled receptors leading to local Ca2+ release in smooth muscle

Membrane depolarization activates voltage-dependent Ca²⁺ channels (VDCCs) inducing Ca²⁺ release via ryanodine receptors (RyRs), which is obligatory for skeletal and cardiac muscle contraction and other physiological responses. However, depolarization-induced Ca²⁺ release and its functional importance as well as underlying signaling mechanisms in smooth muscle cells (SMCs) are largely unknown. Here we report that membrane depolarization can induce RyR-mediated local Ca²⁺ release, leading to a significant increase in the activity of Ca²⁺ sparks and contraction in airway SMCs. The increased Ca²⁺ sparks are independent of VDCCs and the associated extracellular Ca²⁺ influx. This format of local Ca²⁺ release results from a direct activation of G protein-coupled, M₃ muscarinic receptors in the absence of exogenous agonists, which causes activation of Gq proteins and phospholipase C, and generation of inositol 1,4,5-triphosphate (IP₃), inducing initial Ca²⁺ release through IP₃ receptors and then further Ca²⁺ release via RyR2 due to a local Ca²⁺-induced Ca²⁺ release process. These findings demonstrate an important mechanism for Ca²⁺ signaling and attendant physiological function in SMCs.     

Expression of mRNA encoding G protein-coupled receptors involved in congestive heart failure

Congestive heart failure (CHF) induces changes in the neurohumoral system and gene expression in viable myocardium. Several of these genes encode G protein-coupled receptors (GPCRs) involved in mechanisms which compensate for impaired myocardial function. We used real-time quantitative RT-PCR (Q-RT-PCR) to investigate the expression of mRNA encoding 15 different GPCRs possibly involved in CHF, and the effect of normalisation to GAPDH mRNA (GAPDH) or 18S rRNA (18S). CHF was induced in rats by coronary artery ligation, with sham-operated controls (Sham). After 6 weeks, mRNA expression in viable left ventricular myocardium was determined using both 18S and GAPDH as the normalisation standard. An apparent 30% reduction in GAPDH mRNA levels vs. 18S in CHF compared to Sham, although not significant in itself, influenced the interpretation of regulation of other genes.Thus, levels of mRNA encoding receptors for angiotensin II (AT₁), endothelin (ET_A, ET_B) and the muscarinic acetylcholine (mACh) receptor M₁ increased significantly in CHF only when normalised to GAPDH. Levels of mRNA encoding the mACh receptors M₃ and M₄ and the serotonin receptors 5-HT_2A and 5-HT₄ increased, whereas α_1D-adrenoceptor mRNA decreased in CHF irrespective of the normalisation standard. No significant change was detected for M2 and M5 mACh receptors or α_1A-, α_1B-, β₁- or β₂-adrenoceptors. Q-RT-PCR is a sensitive and powerful method to monitor changes in GPCR mRNA expression in CHF. However, the normalisation standard used is important for the interpretation of mRNA regulation. This article is accompanied by the Invited Editorial "Pitfalls in the normalization of real-time polymerase chain reaction data" by M. C. Hendriks-Balk et al. which can be found under     

Acute stress enhances glutamatergic transmission in prefrontal cortex and facilitates working memory

The prefrontal cortex (PFC), a key brain region controlling cognition and emotion, is strongly influenced by stress. While chronic stress often produces detrimental effects on these measures, acute stress has been shown to enhance learning and memory, predominantly through the action of corticosteroid stress hormones. We used a combination of electrophysiological, biochemical, and behavioral approaches in an effort to identify the cellular targets of acute stress. We found that behavioral stressors in vivo cause a long-lasting potentiation of NMDAR- and AMPAR-mediated synaptic currents via glucocorticoid receptors (GRs) selectively in PFC pyramidal neurons. This effect is accompanied by increased surface expression of NMDAR and AMPAR subunits in acutely stressed animals. Furthermore, behavioral tests indicate that working memory, a key function relying on recurrent excitation within networks of PFC neurons, is enhanced by acute stress via a GR-dependent mechanism. These results have identified a form of long-term potentiation of synaptic transmission induced by natural stimuli in vivo, providing a potential molecular and cellular mechanism for the beneficial effects of acute stress on cognitive processes subserved by PFC.     

肠黏膜雌孕激素受体表达在肠易激综合征发病机制中的作用研究

目的 探讨雌、孕激素受体在肠易激综合征（IBS）患者肠道内分布、变化及其临床意义，并对在消化道的作用靶点和可能的细胞学机制做出合理解释和推断。方法 经结肠镜钳取24名正常人和59例肠易激综合征患者的回肠末端、盲肠和降结肠黏膜，分别采用抗人雌、孕激素受体抗体标识肠黏膜上雌、孕激素受体，并应用免疫组化方法检测了雌、孕激素受体阳性细胞数目改变。结果 1、IBS患者回肠末端和盲肠黏膜雌激素受体阳性细胞数目增多（P＜0.01)，降结肠黏膜与正常组无显著差别；2、肠道黏膜未见孕激素受体表达 结论 IBS患者肠黏膜雌激素受体高表达在IBS病理过程中发挥一定作用，并认为肥大细胞为雌激素在消化道的重要靶点，其对于揭示IBS在女性中的高发病率和月经期加重的临床现象有重要意义；同时提示雌激素受体拮抗剂可能对部分患者有效。     

GnRH agonist-induced inhibitory and stimulatory effects during ovarian follicular maturation

The in vivo regulation of ovarian gonadotropin and prolactin receptors and adenylate cyclase activity by FSH, and the potent GnRH agonist [D-Ala⁶]des-Gly¹⁰-GnRH N-ethylamide (GnRHa), was studied in immature hypophysectomized diethylstilbestrol-implanted rats. During FSH treatment over a 48 h period, FSH receptors increased 2-fold with the maximum response during the first 12 h, whereas LH and prolactin receptors increased by 10-fold and 6-fold with the maximum response from 12 to 48 h. Administration of GnRHa at any time during the 48 h period of FSH treatment inhibited the subsequent development of gonadotropin and PRL receptors. In contrast, administration of a single dose of 10 μg GnRHa after 48 h of FSH treatment stimulated follicular luteinization and caused increases in basal adenylate cyclase activity, ovarian weight and PRL receptor content, and concomitant decreases in gonadotropin receptors and adenylate cyclase responses. In the immature follicles of animals not primed with FSH, GnRHa caused progressive inhibition of FSH-sensitive adenylate cyclase activity, with a decrease in FSH receptors, but increased both basal and GMP-P(NH)P-stimulated adenylate cyclase activities. These results demonstrate that GnRHa causes marked inhibition of gonadotropin receptor expression in the basal and FSH-stimulated ovary. This decrease in gonadotropin receptors is an important component of the mechanism by which GnRH agonists inhibit ovarian gonadotropin-sensitive adenylate cyclase activity. In addition, these peptides exert stimulatory effects upon ovarian weight and basal adenylate cyclase activity, and cause an increase in PRL receptors and luteinization of mature ovarian follicles. The stimulatory actions of GnRH agonists upon ovarian function are more prominent with increasing maturation of the granulosa cell.     

Estrogen and progestin receptors in colonic cancer?

Adenocarcinomas of the large bowel in 28 consecutive patients were examined for the presence of estrogen and progestin receptor proteins. None of the specimens showed specific high affinity receptor binding. Our findings suggest that adenocarcinoma of the large bowel does not contain cytoplasmic receptor sites for estrogen and progestin. No reprints will be available.     

Characterization of CD56-/CD16+ natural killer (NK) cells: a highly dysfunctional NK subset expanded in HIV-infected viremic individuals

Natural killer (NK) cells are an important component of the innate immune response against viral infections. NK cell-mediated cytolytic activity is defective in HIV-infected individuals with high levels of viral replication. In the present study, we examined the phenotypic and functional characteristics of an unusual CD56(-)/CD16(+) (CD56(-)) NK subset that is greatly expanded in HIV-viremic individuals. The higher level of expression of inhibitory NK receptors and the lower level of expression of natural cytotoxicity receptors observed in the CD56(-) NK fraction compared with that of CD56(+) NK cells was associated with extremely poor in vitro cytotoxic function of this subset. In addition, the secretion of certain cytokines known to be important in initiating antiviral immune responses was markedly reduced in the CD56(-), as compared with the CD56(+) NK cell subset. These data suggest that the expansion of this highly dysfunctional CD56(-) NK cell subset in HIV-viremic individuals largely accounts for the impaired function of the total NK cell population.     

Steroid receptors in tumors of tissues generally considered to be hormone-independent

Summary One hundred twenty malignant tumors of organs other than breast and uterus were examined for the presence and the amount of steroid receptors in both men and women. Estradiol receptors were most frequently encountered (61%) independent of sex and type of organ. Progesterone receptors were more frequent in renal cell carcinomas than in intestinal carcinomas. Binding sites for dihydrotestosterone and cortisol could be found only occasionally.Supported by a grant of the Deutsche Forschungsgemeinschaft, SFB 118, Grundlagen der Früherkennung und der Verlaufsbeobachtung des Krebses.