Reactivity of ACTH and synthetic ACTH peptides with antisera to human calcitonin.

We studied reactivity of highly purified pituitary hormones in our human calcitonin (hCT) radioimmunoassay (RIA) which can detect 1 pg of hCT. ACTH at doses of greater than 1 microgram of peptide per RIA tube reacted in the hCT assay, as did beta-endorphin (beta EPH) at a dose of 10 micrograms per tube. No reactivity was observed with comparable concentrations of all other known pituitary hormones. ACTH also reacted at doses greater than 1 microgram per tube with 7 other hCT antisera which recognized differing antigenic determinants in the calcitonin molecule but it was not reactive with 2 antisera against porcine calcitonin or 2 antisera against salmon calcitonin. This slight degree of cross-reactivity of hACTH and beta EPH in the hCT RIA cannot account for the presence of immunoreactive CT in pituitary glands. Nevertheless, antisera used for the localization of peptides must be rigorously tested for the existence of cross-reactivities with other possible substances, especially if such antisera detect the peptide in unexpected tissues.     

Fine-needle biopsy of parathyroid adenomas

Summary High-resolution real-time sonography was performed in 15 cases of clinically and chemically suspected primary hyperparathyroidism and in 20 patients with different thyroid nodules. The suspected enlarged parathyroid glands and the thyroid nodules were percutaneously punctured under sonographic control. Concentrations of parathy-roid hormone, human thyroglobulin, and human calcitonin were measured in the aspirate, and immunocytology was performed. The mean concentration of the aspirated parathyroid hormone in the parathyroid glands was 4,013.6 pmol/1±4,519 (SD) as compared with 14.9 pmol/1±8.7 in the thyroid nodules. Thyroglobulin was present in the aspirated fluid of parathyroid adenomas located behind the thyroid (mean±SD, 398.1 ng/ml±317). In comparison, the aspirated thyroglobulin from the thyroid nodules averaged 9,689.7 ng/ml ±3,732. Immunocytology for parathyroid hormone was positive in 14 of the 15 biopsied specimens. Of 15 patients who were scanned for suspected hyperparathyroidism, six had concomitant thyroid nodules.It is concluded that the measurement of high concentrations of parathyroid hormone in the aspirate from a cervical mass, with sonographic control of needle position and/or positive immunocytology provides absolute localization of parathyroid tissue.Abbreviations (FITC) Fluorescein-isothiocyanate - (hCT) human calcitonin - (PBS) phosphate buffered saline - (PTH) parathyroid hormone - (SD) standard deviation - (TG) human thyroglobulin     

高活力人降钙素类似物vhCT的免疫原性研究

本研究用固相多肽合成法合成人降钙素类似物vhCT ,产物纯度达到等电聚焦均一 ,质谱测定分子量精确。生物活性为天然人降钙素的 2 0～ 2 5倍。用放射免疫法测定vhCT能与识别天然人降钙素的抗体起反应 ,产生IgG抗体的免疫原性随偶联的不同载体而有差异。但在BALB/C5 7小鼠体内不引起IgE增高。提示vhCT不产生IgE介导的过敏反应 ,具有药物应用前景。     

Human anti-interleukin 1 alpha antibodies

Autoantibodies to the cytokine interleukin (IL)-1 alpha are frequently found in sera of apparently healthy humans. We have developed a sensitive radioimmunoassay (RIA) and an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of human serum antibodies to IL-1 alpha at concentrations below 10 pmol/l. The RIA is based on coprecipitation of 125I-labelled human recombinant IL-1 alpha (rIL-1 alpha) by rabbit antibodies to human immunoglobulins. The ELISA is based on recovery of added rIL-1 alpha to serum samples and takes advantage of the fact that free human autoantibodies to IL-1 alpha in a dose dependent manner reduce recovery of added rIL-1 alpha. The assays correlate exceedingly well (r = 0.99, P less than 0.001). Their inter- and intraassay coefficients of variation were less than 30% and less than 5% (RIA) and less than 20% and less than 10% (ELISA). Both assays were used to measure the presence of anti-IL-1 alpha antibodies in sera of patients with various autoimmune diseases. Autoantibodies to IL-1 alpha were detectable in up to 75% of these sera, but the frequencies and titers varied considerably between individuals with different diseases.     

Levels of parathyroid hormone-related protein in hypercalcemia of malignancy: comparison of midregional radioimmunoassay and two-site immunoradiometric assay

Summary Overproduction of parathyroid hormone-related protein (PTHrP) is a major cause of hypercalcemia of malignancy in patients with solid tumors. We measured plasma levels of the protein by a radioimmunoassay (RIA) against PTHrP(5384) and by an immunoradiometric assay (IRMA) against PTHrP(1–86). Of 16 affected patients 7 had elevated PTHrP levels in both assays and 4 had elevated levels in the RIA only. Median levels were about tenfold higher in these patients when measured by RIA (median of 34 versus 2.2 pmol/1). Measurements from both assays were, however, highly correlated with each other in this patient group (P<0.01). PTHrP was not elevated in 10 normocalcemic patients with lung carcinoma. During long-term follow-up of a patient with a mesothelioma of the pleura, PTHrP levels measured with both assays decreased during chemotherapy in parallel with a normalization of serum calcium. In another hypercalcemic patient suffering from renal carcinoma, PTHrP measured by IRMA decreased by 40% within 12 h after nephrectomy, whereas PTHrP measured by RIA did not show a significant decline. Direct comparison of the assay results thus pointed to the existence of heterogeneity of circulating forms of PTHrP in plasma. In conclusion, both immunoassays detected elevated levels of PTHrP in a fraction of patients with hypercalcemia of malignancy and thus may be a tumor marker during treatment of malignancies.Abbreviations PTHrP parathyroid hormone-related protein - PTH parathyroid hormone - RIA radioimmunoassay - IRMA immunoradiometric assay     

A new radioimmunoassay for human alpha atrial natriuretic peptide and its physiological validation

A new sensitive, specific radioimmunoassay for human alpha atrial natriuretic peptide (hANP) and a novel extraction method for hANP have been developed. Antiserum to hANP (Keq = 1.923 x 10(11) l/mol) showed no cross-reactivity with related analogues. Displacement of 50% bound 125I-hANP occurred at 4.7 +/- 0.1 fmol/tube (n = 15). The limit of detection of plasma hANP after extraction of 2 ml plasma with Florisil was 1.2 pmol hANP/litre plasma. The recovery of synthetic hANP from plasma over the range 6.5-162.5 pmol/l was 71.2 +/- 1.9%. Inter- and intra-assay coefficients of variation were 13.1% and 10.1% respectively. Extracted plasma was stored at -80 degrees C without loss in immunoreactivity. The radioimmunoassay was physiologically validated in man by measuring plasma hANP following central volume expansion - (a) plasma hANP rose from 2.5 +/- 0.5 to 4.1 +/- 0.6 pmol/l in 9 normal volunteers after postural change from seated to lying (b) infusion of normal saline (2 litre/2 hour) increased hANP from 1.6 to 4.3 pmol/l (n = 2). Infusion of hANP (2 pmol/kg/min) increased plasma hANP to 19.9 +/- 3.4 pmol/l (n = 6).     

Evidence for interference of 25 (OH) vitamin D3 with phosphaturic action of calcitonin.

The present study evaluated the effect of 25(OH)vitamin D3[25(OH)vit D3] on the phosphaturic action of calcitonin in anesthetized parathyroidectomized rats. In group 1, calcitonin was given intravenously over six clearance periods. In group 2, after three periods of calcitonin given intravenously, 25(OH)vit D3 was added and given together with calcitonin for three additional periods. During calcitonin infusion, Cp/CIn 0.18 +/- 0.02 (mean +/- SE) in group 1 was not different from the corresponding Cp/CIn 0.18 +/- 0.03 in group 2; but when 25(OH)vit D3 was added, Cp/CIn 0.12 +/- 0.01 in group 2 was lower (P less than 0.001) than the corresponding Cp/CIn 0.26 +/- 0.02 in group 1. With intravenous calcitonin the urinary excretion of cycle AMP (UcAMP) 97 +/- 29 in group 1 did not differ from the corresponding UcAMP 86 +/- 27 pmol/min in group 2, but when 25(OH)vit D3 was added UcAMP 41 +/- 12 in group 2 was lower (P less than 0.001) than the corresponding UcAMP 131 +/- 14 pmol/min in group 1. This study demonstrated that 25(OH)vit D3 blocks the phosphaturic action of calcitonin. The associated fall in Uctamp suggests that25 (OV)vit D3 acts possibly by inhibiting the calcitonin-induced activation of adenylate cyclase in the kidney. However, other alternative mechanisms of action have not been excluded.     

A simple and sensitive radioimmunoassay for adenosine.

We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2', 3'-0-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[125I]-TME) as a tracer. Anti-adenosine antiserum for the RIA was raised in rabbits immunized with diSc-adenosine conjugated to human serum albumin (diSc-adenosine-HSA). All samples and standards were succinylated prior to assay. The present immunoassay allows detection of 6.25-400 pmol/ml of adenosine in sample. Values obtained by the RIA and by a HPLC analysis showed a high correlation with correlation coefficient of 0.997. In order to determine adenosine in plasmas, blood cells must be separated in the presence of 6 mM EDTA, 0.006% dipyridamole (Dip) and 23 microM 2'-deoxycoformycin (dCF) at 2 degrees C. Adenosine in plasma could be accurately determined by the proposed method even without any pretreatments by deproteinizing. The adenosine levels with or without EDTA-treated normal human plasmas determined were 26.2 +/- 7.26 and 100 +/- 3.62 pmol/ml (mean +/- SEM), respectively.     

妊娠高血压患者血浆中降钙素基因相关肽的测定及其临床意义

本文测定了３０例正常育龄妇女，２８例正常妊娠以及３６例妊娠征患者血浆中降钙素基因相关肽的水平，结果发现，轻度妊高生与正常妊娠相比，血浆中ＣＧＲＰＫ是有下降但不显著（Ｐ〉０．０５）。中重度妊高征与正常妊娠相比，差异有非常显著性，因此推测，妊娠状态下血浆中降钙素基因相关肽的减少可能是妊娠高血压症产生的重要原因之一。     

Sensitive and specific monoclonal immunoassay for detecting yellow fever virus in laboratory and clinical specimens.

A solid-phase radioimmunoassay (RIA) was developed for the detection of yellow fever (YF) virus in infected cell culture supernatant fluid and clinical samples. The test employed a flavivirus group-reactive monoclonal antibody attached to a polystyrene bead support and a radiolabeled type-specific antibody probe in a simultaneous sandwich RIA format. Optimal assay conditions specified a 16-h incubation at high temperature (45 degrees C). Monoclonal antibody to tetanus toxoid was added to the radiolabeled probe to inhibit nonspecific binding. The sensitivity of the assay for cell culture-propagated virus was 2.0 log10 50% mosquito infectious doses per 100 microliters or 100 pg of gradient-purified virion protein per 100 microliters. Specificity, assessed with human sera from 512 patients with liver diseases other than YF, including acute viral hepatitis, showed a false-positive rate of 0.0 to 0.6%. Sera from experimentally infected rhesus macaques containing greater than 3.0 log10 units/100 microliter of YF virus were positive by RIA. Sera and liver tissue from human patients were found to be positive.     

Gamma 2-MSH increases during graded exercise in healthy subjects: comparison with plasma catecholamines, neuropeptides, aldosterone and renin activity

To evaluate the possibility that the proopiomelanocortin (POMC)-derived peptide gamma 2-melanocyte stimulating hormone (gamma 2-MSH) has a role in circulatory regulation in man we studied circulating levels of this peptide at three different stages of physical activity in 10 young healthy subjects. The results were compared to simultaneously measured plasma levels of catecholamines, neuropeptide Y, vasopressin, renin activity, aldosterone and human alpha-atrial natriuretic peptide (alpha-hANP) and of the vasodilatory peptides calcitonin gene-related peptide, substance P and vasoactive intestinal peptide. The plasma levels of gamma 2-MSH-LI (like immunoreactivity) increased from 1009 +/- 101 pmol l-1 at supine rest to 1281 +/- 79 pmol l-1 when measured after 10 min walking (P less than 0.05), and remained at this increased level also after a consecutive further increase of physical activity (4 min stair rush), 1293 +/- 87 pmol l-1 (P less than 0.05 vs. at rest). The increase in circulating gamma 2-MSH-LI levels preceded the elevation of the venous plasma noradrenaline level, but did not rise further with more pronounced activation of the sympathetic nervous system at the highest grade of physical activity examined.     

脑出血患者血浆ET-1、CGRP和NSE变化及临床应用

目的:观察脑出血患者急性发作期、恢复期血浆内皮素(ET-1)、降钙素基因相关肽(CGRP)和神经元特异性烯醇化酶(NSE)的含量变化,探讨它们在脑出血不同时期的临床意义.方法:脑出血组60例,健康对照组60例.脑出血组分别于入院次日、第7d、第21d空腹静脉采集血液标本.用放射免疫分析ET-1、CGRP、NSE含量.结果:脑出血组各时间的ET-1、CGRP和NSE均高于健康对照组,与对照组比较,差异有统计学意义(P＜0.01).ET-1、CGRP发病次日与第7d含量比较,差异无统计学意义(P＞0.05),与第21d含量比较,差异有统计学意义(P＜0.01);NSE发病次日含量与第7d、21d比较,差异有统计学意义(P＜0.01),第7d与第21d含量比较,无统计学意义(P＞0.05).结论:脑出血患者血浆ET-1、CGRP和NSE水平发病次日显著升高.恢复期ET-1、CGRP、NSE含量较发病次日逐渐下降 .测定血浆ET-1、CGRP和NSE含量有助于脑出血患者的早期诊断和预后判断,具有重要的临床价值.     

毛细支气管炎患儿血浆CGRP、VIP RIA的意义

目的：通过愉洲毛细支气管炎患儿急性期和恢复期血浆中降钙素基因相关肽（CCRP）、血管活性肠肽（VIP）的含量,探讨毛细支气管炎患儿血浆CGRP、VIP放射免疫分析（RIA）的意义。方法：本文采用放射免疫分析测定了31例毛细支气管炎患儿急性期和恢复期血浆CGRP、VIP的浓度,以35例正常婴幼儿血浆CGRP、VIP的浓度为对照。结果：毛细支气管炎患儿急性期血浆CGRP浓度明显高于恢复期及正常对照组婴幼儿（P〈0．05）;恢复期CGRP浓度有所下降,但仍高于正常对照组（P〈0．05）,三组间有显著差异（F=82．50,P〈0．01）;毛细支气管炎患儿急性期血浆VIP浓度明显低于恢复期及正常对照组婴幼儿（P〈0．05）;恢复期V1P浓度有所升高,但仍低于正常对照组（P〈0．05）;三组之间有显著性差异（F=300．20,P〈0．01）。结论：CGRP、VIP定量分析结果表明毛细支气管炎患儿血浆CGRP、VIP含量与病程紧密相关。CGRP、VIPRIA对于小儿毛细支气管炎发病机制的研究以及了解其发病病程和指导治疗有着重要的意义。     

中西医结合治疗DVT过程中血浆CGRP水平

目的 :探讨急性深静脉血栓形成 (DVT)中西医结合治疗过程中血浆降钙素基因相关肽 (CGRP)水平及其临床意义。方法 :选择 5 0例急性深静脉血栓形成及 30例本病后遗症接受中西医结合治疗的病人 ,利用放射免疫分析动态观察其血浆CGRP水平。结果 :DVT患者中西医结合治疗后 6h血浆CGRP即明显升高 ,72h达高峰 ,与临床疗效最佳时间相符 ,而 30例后遗症病人则无明显变化。结论 :血浆CGRP水平可作为中西医结合临床治疗效果的一种检测指标 ,并可作为疾病进程的实验室指标。     

C型利钠肽放免试剂盒研制及初步临床应用

目的 :建立C型利钠肽 (CNP)放射免疫分析 ,并初步探讨其临床应用价值。方法 :将甲状腺球蛋白与CNP联接 ,免疫家兔制备抗体。建成CNP放射免疫分析法 ,研制成功放免试剂盒。应用本试剂盒测定83例健康人、5 4例冠心病、2 5例心力衰竭、73例高血压患者血浆CNP水平。结果 :分析灵敏度为 5pg/ml。所获CNP抗血清特异性强 ,与心钠素 (ANP)、神经降压素 (NT)、神经肽Y(NPY)、内皮素 (ET)、降钙素基因相关肽(CGRP)、降钙素 (CT)等多肽均无交叉反应。批内变异系数 <10 % ,批间变异系数 <15 %。心力衰竭、冠心病、高血压患者血浆CNP浓度分别为 6 2 5pg/ml、4 7 9pg/ml、5 6 4pg/ml,明显高于健康人 (2 7 5pg/ml)。结论 :CNP放射免疫分析操作简便、快速 ,为进一步研究CNP在人体生理、病理状态及基础医学研究工作提供了一种可靠的手段。     

A specific, sensitive radioimmunoassay for platelet-activating factor (PAF)

A specific radioimmunoassay (RIA) has been developed for platelet-activating factor (PAF) and shown to be sensitive over the range 10-1000 pg (0.02-2 pmol). The anti-PAF antibodies showed specificity for the acetyl group at the C2 position of the PAF molecule and exhibited no significant cross-reactivity with lyso-PAF or the naturally occurring lipids including lecithin and lysolecithin. The sensitivity of the RIA was at least as good as the platelet-based assays for PAF but the RIA was simpler to perform, had a higher capacity and did not have the drawback of the inherent variability associated with the bioassays.     

成人原发性甲减患者血浆SS、SP含量的变化

目的:观察成人原发性甲状腺机能减退(甲减)患者甲状腺素(TH)治疗前后脑神经肽变化,以探索甲减致脑中枢神经系统损害的机制。方法:采用放射免疫分析,对比分析了45例成人原发性甲减患者TH治疗前后血浆中生长抑素(SS)、P物质(SP)变化。结果:甲减组血浆中SP水平明显低于正常对照组(P<0.01),SS水平与正常对照组比较无显著性差异(P>0.05),其中FT3≥2.5pmol/L组病人血浆中SS水平与正常对照组比较无显著性差异(P>0.05),FT3<2.5pmol/L组血浆中SS含量低于正常对照组(P<0.05),经过TH治疗后SS、SP水平提高(P<0.05,P<0.01)。结论:成人原发性甲减患者血浆存在不同程度的SS、SP减低,这与其引起的神经精神症状可能密切相关,早期、积极、正规的TH治疗对脑功能的恢复有很重要的意义。     

Measurement of glucagon in human plasma by enzyme immunoassay

In order to investigate the validity of an enzyme immunoassay for glucagon, the glucagon levels of human plasma were determined by both enzyme immunoassay (EIA) and radioimmunoassay (RIA). After a glucose load, plasma glucagon measured by both EIA and RIA fell in 12 normal subjects. The glucagon levels measured by both assays during glucose tolerance test showed good agreement in a group of 10 patients. After arginine infusion, plasma glucagon increased in 6 normal subjects and 3 patients and glucagon values measured by EIA correlated well with those by RIA. The present study demonstrates correlation between glucagon levels measured by RIA and EIA and indicates the usefulness of EIA for determining glucagon in human plasma.     

Vasodilatory effects and coexistence of calcitonin gene-related peptide (CGRP) and substance P in sensory nerves of cat dental pulp

Substance P (SP)- and calcitonin gene-related peptide (CGRP)-immunoreactivity (-IR) were localized by immunohistochemistry in the same nerve cell bodies in the trigeminal ganglia as well as in nerve terminals of the dental pulp. The distribution of SP- and CGRP-IR nerves were identical in the dental pulp and mainly associated with blood vessels. The level of CGRP-IR in the dental pulp, as measured by radio-immunoassay (RIA), was 1.4 +/- 0.2 pmol g-1 wet wt, which is in the same range as that found for substance P. Local intra-arterial infusion of synthetic CGRP and substance P produced vasodilatation in the dental pulp as measured by both laser Doppler flowmetry and an 125I clearance technique. The CGRP was effective as a vasodilator when infused in the femtomole per minute range, and SP in the picomole range. The effect of CGRP (50 fmol min-1) was 10 times larger when given after SP (15 pmol min-1) than before it. Since the two peptides coexist in the same neurons, it is suggested that they both contribute to the vasodilation seen upon antidromic stimulation of sensory nerves.     

缺氧缺血性脑病新生儿血浆ET与CGRP含量测定的临床意义

为探讨内皮素（ＥＴ）与降钙素基因相关肽（ＣＧＲＰ）在新生儿缺氧缺血性脑病（ＨＩＥ）中的临床意义,采用放射免疫分析法测定了３５例ＨＩＥ新生儿血浆ＥＴ与ＣＧＲＰ含量变化。结果显示,ＨＩＥ新生儿急性期血浆ＥＴ水平与对照组比较显著升高（Ｐ〈０．０１）,恢复期较急性期明显降低（Ｐ〈０．００１）,但仍明显高于对照组（Ｐ〈０．００１）;急性期血浆ＣＧＲＰ含量较对照组明显降低（Ｐ〈０．００１）,恢复期较急性期显著