La imagen nuclear con anticuerpos monoclonales radiomarcados: viejas aplicaciones con nuevos enfoques / Nuclear imaging with radiolabeled monoclonal antibodies: old applications with new aproaches

Resumen Los anticuerpos monoclonales marcados con radionucleidos fueron en sus inicios ampliamente empleados para el estudio de diversas enfermedades, fundamentalmente oncológicas mediante la inmunogammagrafía. Estos fueron poco a poco sustituidos por moléculas con mejores prestaciones como los péptidos y la 18F-fluordesoxiglucosa (18F-FDG). No obstante, en el presente siglo, la amplia introducción de la inmunoterapia produjo un cambio de paradigma en cuanto al empleo de los anticuerpos monoclonales radiomarcados para la adecuada selección y seguimiento de los pacientes a ser tratados con inmunoterapia, resurgiendo la inmunotomografía por emisión de fótón único (inmuno-SPECT), la inmunotomografía por emisión de positrones (inmuno-PET) y la imagen corregistrada con la tomografía axial computarizada (TAC), como modalidades de gran valor en el manejo del cáncer. El objetivo del presente trabajo fue brindar una panorámica acerca de la evolución de la imagen nuclear con anticuerpos monoclonales radiomarcados y sus principales aplicaciones en el tiempo, fundamentalmente en el estudio de los pacientes con cáncer.

Abstract In the beginning, radionuclide-labeled monoclonal antibodies were widely employed for the study of various diseases, mainly oncological, by immunoscintigraphy. They were gradually replaced by molecules with better performance such as peptides and 18F-FDG. However, in the present century, the wide introduction of immunotherapy produced a paradigm shift in the use of radiolabeled monoclonal antibodies for the proper selection and follow-up of patients to be treated with immunotherapy, re-emerging of the immune-single photon emission tomography (immuno-SPECT), the immune-positron emission tomography (immuno-PET) and the co-registered image with computed tomography (CT) as imaging modalities of great value in the management of cancer. The aim of the present work was to provide an overview of the evolution of nuclear imaging with radiolabeled monoclonal antibodies and their main applications over the time, mainly in the study of patients with cancer.

Servicio de determinación de hormonas a instituciones hospitalarias en el Centro de Isótopos / Hormone determination service to hospital institutions the Center of Isotopes

RESUMEN En los marcos de la estrecha colaboración entre la Agencia de Energía Nuclear y Tecnologías de Avanzada y el Ministerio de Salud Pública, a inicios de este siglo se procedió a organizar, en el Centro de Isótopos, un servicio de determinación de hormonas por metodologías nucleares, atendiendo a necesidades de diversas instalaciones hospitalarias. El centro disponía de personal capacitado en técnicas de radioinmunoanálisis y seguridad radiológica, el equipamiento necesario y laboratorios licenciados para el trabajo con sustancias radiactivas. Para ello se hizo necesario incorporar al Sistema de Calidad las regulaciones vigentes para laboratorios clínicos y adecuar la infraestructura para el procesamiento de no menos de 1000 muestras por semana, procedentes de 16 hospitales. Se requería recogida y transportación de las muestras, recepción, inspección y registro, análisis, evaluación y entrega de los resultados. En función de la actividad se diseñó y puso en operación un sistema informático y se normalizaron acciones para asegurar competencia técnica avalada por: trazabilidad; incertidumbre; ejercicios de aptitud y otros requisitos exigidos por la regulación de Buenas Prácticas de Laboratorio Clínico vigentes en el país. A raíz de nuevas regulaciones, el servicio ha introducido de manera paulatina determinadas mejoras. En 15 años de trabajo este laboratorio ha resultado el principal del país en emplear métodos radiactivos para la cuantificación de hormonas, lo que favorece la atención de más de 70 000 pacientes como promedio al año, principalmente de la capital. Esto es parte de nuestra contribución en el 500 aniversario de La Habana.

ABSTRACT Within the framework of the close collaboration between the Agency of Nuclear Energy and Advanced Technologies and the Ministry of Public Health, at the beginning of this century, and considering the need of different hospitals, the Center of Isotopes (CENTIS), began providing service to determine hormones by nuclear methodologies,. CENTIS had qualified personnel in immune radioanalysis RIA/IRMA techniques and radiation safety, in addition to having the required equipment and accredited laboratories to work with radioactive substances. As a result, it was necessary to incorporate to CENTIS Quality System, the regulations in force for clinical laboratories , and to adapt its infrastructure for the processing of no less than 1000 samples per week from 16 hospitals. The Service required collection and transportation of the samples, reception, inspection and registration, test, evaluation and delivery of the test results. In order to fulfil this activity, a computer system was designed and set , and standards for actions were established to guarantee a high technical quality endorsed by traceability, uncertainty; proficiency tests and other requirements demanded by the Good Practices of Clinical Laboratory (BPLC 03 -2009) issued by CECMED. Due to new regulations, several improvements have been introduced. In 15 years, CENTIS laboratory has become the most important in the country using radioactive methods for the quantification of hormones, with an average testing of more than 70 000 patients per year, mainly in the capital. This is part of our contribution in the 500th Anniversary of Havana.

On the consistency of different methods for detecting aldosterone concentration in blood / 中华内分泌代谢杂志

Objective@#To evaluate the consistency of different methods for detecting aldosterone concentration in blood and to establish a reference interval of serum aldosterone concentration by liquid chromatography-tandem mass spectrometry(LC-MS/MS).@*Methods@#Concentrations of blood aldosterone were measured by LC-MS/MS, chemiluminescent assays (Diasorin, Domestic A and B systems) and radioimmunoassay (RIA) in 138 healthy adults, 67 patients with essential hypertension and 23 patients with primary aldosteronism.@*Results@#Aldosterone concentrations measured by various methods were quiet different(P<0.01). Spearman correlation analysis showed that the correlation coefficient were 0.776(P<0.01) between CLIA (Diasorin) and LC-MS/MS, 0.805(P<0.01) between CLIA (Domestic A) and LC-MS/MS, 0.058(P>0.05) between CLIA(Domestic B) and LC-MS/MS, 0.338(P<0.01) as well as between RIA and LC-MS/MS. Bland-Altman analysis for the measurements showed poor consistency among methods for aldosterone concentrations. The reference interval was 15.2-222.2 pg/ml for serum aldosterone concentrations by LC-MS/MS.@*Conclusions@#There are significant differences of blood aldosterone concentrations determined by different methods. Clinically, a specified reference interval might be needed while using different methods.

Research progress of aldosterone detection methods and clinical applications / 中华检验医学杂志

Aldosterone is a steroid hormone that acts on the kidneys to maintain the balance of sodium and water. Primary hyperaldosteronism (PA) is the most common disease that causes secretory hypertension. PA is often unrecognized and is associated with a high death rate. The aldosterone/rein rate (ARR) and detection of serum or urinary aldosterone are the most effective method to screen and identify PA. In healthy individuals, aldosterone concentration is present in low quantities in serum and there are numerous compounds that can potentially interfere with analysis making aldosterone a technically challenging analyte quantifying method. At present, the method of aldosterone detection includes radioimmunoassay, enzyme-linked immunosorbent assay, chemiluminescence method and mass spectrometry. These methods have their own advantages and disadvantages, the ARR ratios of different detection methods for the diagnosis of PA are not completely consistent. In this paper, the detection methods and clinical applications of aldosterone are reviewed.

Research progress of aldosterone detection methods and clinical applications / 中华检验医学杂志

Aldosterone is a steroid hormone that acts on the kidneys to maintain the balance of sodium and water. Primary hyperaldosteronism (PA) is the most common disease that causes secretory hypertension. PA is often unrecognized and is associated with a high death rate. The aldosterone/rein rate (ARR) and detection of serum or urinary aldosterone are the most effective method to screen and identify PA. In healthy individuals, aldosterone concentration is present in low quantities in serum and there are numerous compounds that can potentially interfere with analysis making aldosterone a technically challenging analyte quantifying method. At present, the method of aldosterone detection includes radioimmunoassay, enzyme-linked immunosorbent assay, chemiluminescence method and mass spectrometry. These methods have their own advantages and disadvantages, the ARR ratios of different detection methods for the diagnosis of PA are not completely consistent. In this paper, the detection methods and clinical applications of aldosterone are reviewed.

Application value of anti-double stranded DNA antibody in systemic lupus erythematosus by enzyme-linked immunosorbent assay / 上海交通大学学报（医学版）

Objective • To investigate the characteristics and clinical application value of anti-double stranded DNA (dsDNA) antibody detected by enzyme-linked immunosorbent assay (ELISA). Methods • 186 patients with systemic lupus erythematosus, 183 autoimmune disease of non-SLE controls, 78 non-autoimmune disease controls and 50 healthy controls were selected. The serum anti-dsDNA antibody was detected simultaneously by the methods of ELISA and radioimmunoassay (RIA) and their diagnostic efficacies for detection were compared. Results • The sensitivities of anti-dsDNA antibody in SLE by RIA and ELISA were 47.31% and 62.90%, respectively. The specificities were 85.85% and 81.67%, respectively. The positive predictive were 66.67% and 67.24%, respectively. The negative predictive were 73.15% and 78.14%, respectively. The anti-dsDNA antibody levels of SLE patients detected by ELISA and RIA both increased with the increase of SLE disease activity index. Conclusion • The specificity of ELISA is similar with RIA in diagnosing SLE, and the sensitivity is higher than RIA, which can screen the patients with SLE. In addition, the two methods are both suitable for monitoring the condition of SLE.

Survey Result of the Counting Efficiency of Gamma Counter by Certified Reference Materials / 대한핵의학회잡지

PURPOSE: In radioimmunoassay (RIA), the gamma counter is the important instrument for the accurate measurement. To manage quality assurance of RIA, the counting efficiency of gamma counter is one of the important parameters. The aimof this study was to evaluate the counting efficiency of gamma counters in multiple institutes on the base of traceability by using the certified reference materials (CRMs).METHODS: Twenty-three institutes that perform RIA were enrolled in this study. I-125 CRMs that were certified by National Institute of Standards and Technology (NIST) were used. Each institute was asked to count the activity of I-125 CRMs at most twice on all gamma counters in use. The counting efficiency of each well of counter was calculated on the base of NIST-certified information, corrected for I-125 decay for date of testing.RESULTS: From 23 institutes, 44 gamma counters were evaluated. The average counting efficiency of all wells was 85.9% and the standard deviation was 13.5%. As a mean value of each gamma counter, three gamma counters showed poor counting efficiency (less than 70%). The poorest counting efficiency was 7%. The counting efficiency of seven gamma counters was between 70 and 75%. Eight counters had the counting efficiency between 75 and 90%. More than half of counter (26 gamma counters) showed excellent counting efficiency (more than 90%). The standard deviation variation range of inter-well efficiency was from 0 to 11.2.CONCLUSION: The first survey on the counting efficiency of gamma counter was performed in South Korea. Most of the RIA laboratories have well managed the quality assurance of gamma counter.

Current status and prospects of quantitative immunoassay technologies / 中华检验医学杂志

Quantitative immunoassay technique is the common method of quantitative detection in clinical laboratory. Several important branches of quantitative immunoassay were formed by changing the tracer or the Antigen-antibody complex separation method, including radioimmunoassay, fluorescent immunoassay, enzyme immunoassay, chemiluminescence, colloidal gold, immuno-turbidimetric analysis and homogeneous immunoassay. Different immunoassay techniques have their own characteristics, also apply to different detecting conditions in clinic. This paper reviewed several common kinds of quantitative immunoassay technology, and discussed both their advantages and disadvantages, which provide reference for the application and development of clinical testing technology.

Estradiol and progesterone fecal metabolites analysis in crab-eating-fox (Cerdocyoun thous) / Análise de metabólitos fecais de estradiol e progesterona em cachorro-do-mato (Cerdocyoun thous)

In this study, four crab-eating fox females (Cerdocyon thous) maintained at the Federal University of Mato Grosso Zoo, Cuiabá, Brazil, were investigated for 12 months, using feces measurement of estradiol and progesterone concentrations. Fecal collections were performed three times a week for hormone extraction. Two methods of analysis, Elisa (EIA) and Radioimmunoassay (RIA), were used in the measurement of progesterone (P4) and estradiol (E2) metabolites. The aim of this study was to compare and validate two different methods of hormone measurement for C. thous. There were no differences regarding the method used. The Radioimmunoassay technique proved to be more sensitive, however, both showed similar results.(AU)

Neste estudo, quatro fêmeas de cachorros-do-mato (Cerdocyon thous) mantidas no Zoológico da Universidade Federal de Mato Grosso, Cuiabá, MT, Brasil, foram investigadas pelo período de 12 meses, mediante a mensuração de concentrações de estradiol e progesterona em fezes. Coletas de fezes foram realizadas três vezes por semana para posterior extração hormonal. Dois métodos de análise de metabólitos fecais, elisaimunoensaio (EIA) e radioimunoensaio (RIA), foram utilizados na mensuração dos metabólitos de progesterona (P4) e estradiol (E2). O objetivo deste estudo foi comparar e validar dois diferentes métodos de mensuração hormonal para C. thous. Não houve diferença significativa com relação ao método empregado. A técnica de radioimunoensaio demonstrou ser mais sensível, no entanto ambas apresentaram resultados semelhantes.(AU)

Assessment of the Precision and Functional Sensitivity of Two Thyroglobulin Assays: Comparison of the Second-Generation Roche Electrochemiluminescent Immunoassay and BRAHAMS Radioimmunoassay

BACKGROUND: Thyroglobulin (Tg) is the primary biochemical marker used to monitor patients with differentiated thyroid cancer (DTC) for residual or recurrent disease after total thyroidectomy, as only normal or well-differentiated malignant thyroid cells produce Tg. Here, we evaluated the precision and functional sensitivity (FS) of a recently developed highly sensitive Tg (hsTg) electrochemiluminescent immunoassay (ECLIA) and compared it to that of the radioimmunoassay (RIA) method using pooled human serum with low levels of Tg. METHODS: For the ECLIA method, the Elecsys Tg II kit (Roche Diagnostics, Germany) was used with an E170 analyzer (Roche Diagnostics). For the RIA method, the Tg-plus-RIA kit (BRAHAMS, Germany) was used with a Cobra Quantum gamma counter (Packard Instrument Company, USA). The precision and limit of detection (LOD) were determined according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. FS was determined using a modification of the CLSI guideline. RESULTS: The total precision of the hsTg ECLIA and RIA methods was 9.6% and 48.2%, respectively. The manufacturer-reported LOD was verified by the hsTg ECLIA (0.04 ng/mL), but not by the RIA method (>0.08 ng/mL). The hsTg ECLIA showed better FS (0.04 ng/mL at a coefficient of variation [CV] of 10%) than the RIA method (0.37 ng/mL at a CV of 20%). CONCLUSIONS: Thus, the hsTg ECLIA performed better than the RIA method in terms of FS, which is extremely important for the early detection of residual or recurrent disease in DTC patients after total thyroidectomy. The excellent performance of the hsTg ECLIA could allow for clinical Tg measurement without thyroid-stimulating hormone stimulation, in contrast to the insufficient performance of the RIA method.

Determination of cortisol and cortisone in human saliva by a liquid chromatography-tandem mass spectrometry method / Determinação de cortisol e cortisona na saliva por método baseado em cromatografia líquida e espectrometria de massas em tandem

Objective Salivary cortisol measurement plays an important role in the evaluation of adrenal function. Its high correlation with free serum cortisol, the easy of sampling and the limited presence of interfering steroids, generated multiple recent studies of its application, in special in the screening of adrenal hyperfunction. In this paper we present our experience in the development of a high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for salivary cortisol and cortisone measurement. Materials and methods For this study we used 181 saliva samples from our routine diagnostic laboratory. The HPLC-MS/MS method was based on a Waters Quattro Premier tandem mass spectrometer with an electrospray probe. After derivatization with hydroxylamine transitions monitored included cortisol and cortisone. An in-house radioimmunoassay (RIA) was used for salivary cortisol results comparison. Results Functional sensitivity was 24 ng/dL for cortisol and linearity from 24 to 1929 ng/dL. Saliva cortisol values obtained in the 181 samples presented a median of 52 ng/dL with 5‐95% percentile of 24 and 374 ng/dL. With the RIA the results were 86, 25 and 436 ng/dL, respectively, with values for RIA being significantly higher (P<0.0001) and high correlation (r=0.8312, P<0.0001). Cortisone measured in 159 samples showed a median of 278 ng/dL, with 5‐95% percentile of 100 and 1,133 ng/dL. Correlation with cortisol values was significant (r=0.820, P<0.0001). Conclusion We conclude that the HPLC-MS/MS method compares favorably with the RIA for salivary cortisol measurement, with the additional possibility of concomitant cortisone measurement and the evaluation of 11βHSD2 activity. .

Objetivo A dosagem de cortisol salivar é uma metodologia que vem tendo crescente aceitação no estudo da função adrenocortical. Sua alta correlação com a fração livre sérica, facilidade de coleta e presença limitada de interferentes têm originado múltiplas publicações, em especial no screening de pacientes suspeitos de hiperfunção. Neste trabalho apresentamos nossa experiência no desenvolvimento de metodologia baseada em cromatografia líquida e espectrometria de massas (HPLC-MS/MS) para a medida de cortisol e cortisona salivares. Materiais e métodos Para este estudo utilizamos 181 amostras de saliva de nossa rotina diagnóstica. A metodologia de HPLC-MS/MS baseou-se num espectrômetro de massas Waters Quattro Premier. Após derivatização com hidroxilamina, as transições monitoradas incluíram cortisol e cortisona. Um radioimunoensaio (RIE) in house foi empregado para comparação. Resultados A sensibilidade funcional para cortisol foi de 24 ng/dL, com linearidade entre 24 e 1,929 ng/dL. Os valores de cortisol obtidos nas 181 amostras apresentaram mediana de 52 ng/dL, com percentis 5‐95% de 24 e 374 ng/dL. Com o RIE, os resultados foram 86, 25 e 436 ng/L, respectivamente, com os valores obtidos no RIE significativamente mais elevados (P<0,0001), e alta correlação (r=0,8312, P<0,0001). Cortisona, medida em 159 amostras, mostrou mediana de 278 ng/dL, com percentis 5‐95% entre 100 e 1.133 ng/dL. A correlação com os valores de cortisol foi significativa (r=0,820, P<0,0001). Conclusão Concluímos que o método baseado em HPLC-MS/MS compara-se favoravelmente com o RIE para a medida de cortisol salivar, com a possibilidade adicional da medida concomitante de cortisona e avaliação da atividade da enzima 11βHSD2. .

Análise de um radioimunoensaio iodado para determinação de 11-deoxicortisol / Analysis of an iodide radioimmunoassay for 11-deoxicortisol measurement

Objetivo : Nosso objetivo foi comparar duas técnicas de dosagem do 11-desoxicortisol: a técnica de radioimunoensaio iodado, a qual foi validada neste trabalho, e a cromatografia líquida de alta performance seguida por espectrometria de massa em tandem (LC-MS/MS), sendo a última considerada o padrão-ouro para dosagem dos hormônios esteroides. Materiais e métodos : Para a comparação entre os resultados de 11-desoxicortisol, foram selecionadas 88 amostras. Resultados : A sensibilidade analítica do radioimunoensaio foi de 0,30 ng/mL, com linearidade e perfil de precisão inadequado (34% das amostras com CV ≥ 20%). Das 88 amostras selecionadas, apenas 54 apresentaram resultados mensuráveis em ambos os métodos. A comparação desses resultados, por meio da regressão de Deming, resultou em um coeficiente de correlação de 0,610, inclinação de 3,751, intercepção de 0,145, evidenciando a pobre correlação entre os resultados e a superestimação dos resultados pelo RIA. Conclusão : Concluímos que o método de dosagem de 11-desoxicortisol por radioimunoensaio iodado apresentou resultados inadequados nos diversos parâmetros avaliados, inviabilizando sua utilização como método de dosagem do 11-desoxicortisol. Arq Bras Endocrinol Metab. 2014;58(3):232-6 .

Objective : Our aim was to correlate 11-deoxycortisol levels obtained by two currently available techniques for 11-deoxycortisol measurement: radioimmunoassay, and high performance liquid chromatography followed by tandem mass spectrometry (MS/MS). The latter is the gold standard method for steroid hormone measurement. Materials and methods : We selected 88 samples and the results of these two methods were compared by Deming regression. Results : The analytical sensitivity of the RIA was 0.30 ng/mL, with inadequate linearity and inadequate precision profile (34% of the samples had a CV ≥ 20%). From the selected samples, 54 had measurable levels of 11-deoxycortisol in both methods and were used in the comparison. The comparison of RIA with LC-MS/MS showed an overestimation of the results by RIA. The correlation coefficient was 0.610; linear regression slope was 3.751; and the intercept was 0.145, indicating a poor correlation between the two methods. Conclusion : We concluded that 11-deoxycortisol measured by radioimmunoassay, despite a good analytical sensitivity, showed very low specificity, precluding its use as a reliable method for 11-deoxycortisol measurement. Arq Bras Endocrinol Metab. 2014;58(3):232-6 .

Influence of the treatment of microwave ablation combined with 131Ⅰ tumor cells human mouse chimeric monoclonal antibody injection on the immune function of patients with lung cancer / 肿瘤研究与临床

Objective To investigate the mechanism of anti-tumor in lung cancer patients and their effects to immune system undergoing with microwave ablation treatment,radioimmunotherapy of 131Ⅰ tumor cells human mouse chimeric monoclonal antibody injection (131Ⅰ-chTNT) and combined treatment of two anterior method.Methods The 50 cases of lung cancer were divided into three groups randomly,17 cases were in the group of simple microwave ablation treatment,15 cases were in the group of simple radiotherapy immunotherapy,18 cases were in the group of combined microwave ablation treatment with 131Ⅰ-chTNT radioimmunoassay.During the study,the T lymphocyte subsets,the activity of NK cells,the expressing of interleukin (IL)-2,IL-10,IL-12,the changing of the interferon γ(IFN-γ) and tumor necrosis factor α (TNF-α) were detected and compared of before and after treatment on all the patients.Results The CD4+ value,CD4+/CD8+ value and the activity of NK cells in these three groups after treatment were higher than that of before treatment (P ＜ 0.05),there was statistic difference between the combined treatment group and two simple treatment groups (P ＜ 0.01).The expression of IL-2,IL-12,IFN-γ TNF-α in three group patients after treatment were higher than that of before treatment,which had statistical significance (P ＜ 0.05),while there were no significant differences between groups (P ＞ 0.05).Conclusion The method combined microwave ablation treatment with 131Ⅰ-chTNT radioimmunoassay has efficiency to improve the immune function,which could improve the comprehensive therapeutic effect of lung cancer excellently.

Hormonas reprodcctivas de importancia veterinaria en hembras domésticas rumiantes / Reproductive hrmmones of veterinary importance in domestic ruminant females

En el presente trabajo se revisan las principales hormonas reproductivas en hembras domésticas rumiantes, considerando sus características e interacciones, además de los métodos de determinación y las concentraciones que se reportan de cada una de dichas hormonas, durante las diferentes fases del ciclo estral. El eje hipotálamo-pituitario-ovárico controla la actividad reproductiva, principalmente, a través de las interacciones entre la Hormona Folículo Estimulante (FSH), la Hormona Luteinizante (LH), el Estradiol (E2) y la Progesterona (P4). Durante la fase folicular, las gonadotropinas estimulan el desarrollo de los folículos, promoviendo la proliferación de las células de la granulosa por parte de la FSH; su pico está asociado al surgimiento de la onda folicular, después de la cual decrece la concentración plasmática de FSH, y da inicio a la desviación folicular. Esto permite al folículo dominante expresar receptores para la LH, además de producir inhibina y E2. El alto nivel circulante de E2 induce la liberación de la Hormona Liberadora de Gonadotropinas (GnRH) desde el hipotálamo, resultando en un pico de LH, de amplitud y frecuencia suficiente para estimular la maduración final del folículo y la posterior ovulación. Las altas concentraciones de E2 influyen también sobre la presentación de los cambios fisiológicos y comportamentales durante el estro. La fase luteal está caracterizada por el predominio de la P4, cuya concentración se relaciona con el desarrollo del cuerpo lúteo. Esta hormona es indispensable para el reconocimiento y mantenimiento de la preñez y sus perfiles pueden llegar a determinar si existe la predisposición de un animal a sufrir pérdidas embrionarias tempranas. Se considera que los incrementos o disminuciones en las concentraciones séricas de cada hormona marcan cambios en las fases del ciclo estral, y es fundamental conocer la actividad de dichas hormonas mediante la determinación de su concentración sanguínea normal para cada una de las etapas reproductivas.

The main reproductive hormones in domestic female ruminants are reviewed in this paper, including their characteristics, interactions and concentrations besides the determination methods and concentrations reported in each hormone during the different phases of the estrous cycle. The hypothalamus-pituitary-ovarian axis, controls reproductive activity, mainly, through interactions between the Follicle-stimulating Hormone (FSH), Luteinizing Hormone (LH), Progesterone (P4) and Estradiol (E2). During the follicular phase, gonadotropin hormones stimulate follicle development, promoting the proliferation of granulosa cells on the FSH side. The FSH peak is associated with the emergence of the follicular wave after which, its plasmatic concentration decreases and the follicular deviation begins. This allows the dominant follicle to express LH-receptors and to produce inhibine and E2. The high level of flowing E2 induces the release of Gonadotropin Releasing Hormone (GnRH) from the hypothalamus, resulting in a LH of enough amplitude and frequency to stimulate the final maturation of the follicle and the subsequent ovulation. High plasma concentrations of E2, also influence the appearance of physiological and behavioral changes during estrous. The luteal phase is characterized by the dominance of P4, whose concentration is related to the development of the luteal body. This hormone is essential for pregnancy and its profiles recognition and maintenance, and it can even determine if there exist predisposition of an animal to suffer early embrionary loss. It is considered that increase or decrease in plasma concentration of each hormone, determine changes in the estrous cycle phases and, it is fundamental to know the activity of such hormones, by determining their normal blood concentration for each of the reproductive stages.

Performance Evaluation of Method for Detecting Serum 25-Hydroxyvitamin D2 and 25-Hydroxyvitamin D3 by Using PerkinElmer MSMS Vitamin D Kit / 임상검사와정도관리

BACKGROUND: Determining the concentration profiles of serum 25-hydroxyvitamin D (25OHD) may aid in the clinical diagnosis and treatment of vitamin D deficiency. To date, the standardized liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay has been used for accurate and precise determination of 25-hydroxyvitamin D levels. Here, we evaluated the performance of the recently developed and introduced PerkinElmer Vitamin D kit and compared the measurements obtained by RIA and LC-MS/MS methods. METHODS: We evaluated the accuracy, precision, linearity, lower limit of quantification (LLOQ), recovery, and carry-over of the MSMS Vitamin D kit. Clinical specimens from 80 patients were used for the comparison between the MSMS Vitamin D kit (PerkinElmer, USA) and the RIA kit (DiaSorin, USA). RESULTS: The MSMS Vitamin D kit was found to produce intra-assay and inter-assay coefficients of variation (CVs) of less than 6% for precision and showed a bias of less than 5%. The MSMS Vitamin D kit displayed linearity within the range for total vitamin D levels of 4.5-150 ng/mL, and the lower limit of quantification for 25OHD was 0.38 ng/mL. The RIA measurements of 25OHD showed a correlation of y=0.9931x+0.2216 (r2=0.74) with the LC-MS/MS values. CONCLUSIONS: The LC-MS/MS assay of 25OHD3 and 25OHD2 showed excellent performance when using the MSMS Vitamin D kit and in terms of 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) derivatization. Further, the results obtained were well correlated with those obtained using the RIA method. Thus, assays using the MSMS Vitamin D kit are considered as more standardized, and they enable quicker and more accurate analysis and help reduce inter-laboratory variation than that by other existing methods.

Desenvolvimento de um radioimunoensaio de dexametasona no soro como subsídio para o diagnóstico da síndrome de Cushing / Development of serum dexamethasone radioimmunoassay to corroborate Cushing's syndrome diagnosis

INTRODUÇÃO: O teste de supressão com 1 mg de dexametasona (TSDx) é amplamente empregado no rastreamento da síndrome de Cushing (SC) dada sua elevada acurácia diagnóstica. A SC é um distúrbio endocrinometabólico resultante do hipercortisolismo crônico com característica de ausência de supressão do cortisol no TSDx. OBJETIVO: Desenvolver um radioimunoensaio (RIE) para a dosagem de dexametasona (Dx) no soro para complementar o TSDx. MÉTODOS: Foram imunizados três coelhos com o conjugado dexametasona-21-hemissuccinato-BSA para escolha do melhor anticorpo e o RIE foi desenvolvido de acordo com as recomendações da RE 899/2003 da Agência Nacional de Vigilância Sanitária (ANVISA). Analisamos 96 voluntários, sendo 67 submetidos ao TSDx e 29 não, e 12 pacientes com SC, estudados na Universidade Federal de São Paulo (UNIFESP) e na Santa Casa de Misericórdia de São Paulo. RESULTADOS: O anticorpo contra Dx selecionado mostrou boa especificidade, coeficiente de variação (CV por cento) intra e interensaio < 20 por cento, exatidão de 93,8 por cento e dose mínima detectada de 19,5 ng/dl. A concentração de Dx no soro foi semelhante nos voluntários e pacientes com SC (ausência/supressão do cortisol): 205 a 703 ng/dl e 174 a 661 ng/dl (intervalo de confiança [IC] 95 por cento), respectivamente; os valores foram indetectáveis naqueles que não se submeteram ao teste. DISCUSSÃO: O anticorpo empregado apresenta boa afinidade e especificidade para quantificar a Dx no soro. O RIE mostrou reprodutibilidade e eficiência na determinação dos níveis séricos de Dx durante o TSDx. CONCLUSÃO: O presente RIE para a dosagem de Dx no soro é acurado e confiável, permitindo estabelecer uma faixa de referência de valores para subsidiar a interpretação do TSDx.

INTRODUCTION: The 1 mg dexamethasone suppression test (DxST) is widely used to screen Cushing's syndrome (CS) due to its high diagnostic accuracy. CS is an endocrine-metabolic disorder caused by hypercorticism, which is characterized by the absence of cortisol suppression in DxST. OBJECTIVE: To develop a radioimmunoassay (RIA) for the measurement of serum dexamethasone (Dx) to complement DxST. METHODS: Three rabbits were inoculated with dexamethasone-21-hemisuccinate-BSA in order to choose the best antibody. Serum Dx RIA was performed according to RE 899/2003 (Agência Nacional de Vigilância Sanitária [ANVISA]) regulations. Serum samples from 96 volunteers from Universidade Federal de São Paulo (UNIFESP) and Santa Casa de Misericórdia de São Paulo were analyzed, 67 of which were submitted to DxST and 29 were not. There were 12 patients with CS. RESULTS: The Dx antibody chosen showed good specificity. Intra- and interassay CV were < 20 percent with 93.8 percent accuracy and the lowest detection limit was 19.5 ng/dl. Serum Dx concentration was similar among both volunteers and CS patients (absence of cortisol suppression): 205 to 703 ng/dl and 174 to 661 ng/dl (95 percent CI), respectively. Values were undetectable among those that were not submitted to the test. Discussion: The anti-Dx antibody shows high specificity and reliability to quantify serum Dx in DxST. The Dx RIA presented reproducibility and reliability in the determination of serum Dx levels during DxST. CONCLUSION: The current RIA for serum Dx is accurate and reliable, which permits to establish a reference value range to substantiate DxST interpretation.

Combination of IGF-1 with CEA, CYFRA21-1, NSE for the diagnosis and prediction of treatment response in lung cancer / 中华核医学杂志

Objective To evaluate four tumor markers of insulin-like growth factor 1((IGF-1), CEA, cytokeratin fragment antigen 21-1 (CYFRA21-1), neuron-specific enolase (NSE) for the diagnosis and prediction of treatment response in human lung cancer. Methods Serum samples were taken from three groups: 91 patients with lung cancer, 30 healthy adults and 15 patients with benign pulmonary diseases. Serum IGF-1 was assayed by radioimmunoassay and CEA, CYFRA21-1, and NSE by electrochemiluminescence immunoassay. The differences among the three groups were determined by Kruskal-Wallis one-way analysis of variance (ANOVA) and with Mann-Whitney rank-sum test. Diagnostic efficacy was evaluated by ROC curves. Results The four serum tumor marker levels were significantly higher in lung cancer group, as compared with the benign and the healthy (IGF-1:χ2=26.95,P<0.001, CEA:χ2=49.11,P<0.001; CYFRA21-1:χ2=40.63,P<0.001; NSE:χ2=14.76;P<0.001). The diagnostic sensitivities of IGF-1, CEA, CYFRA21-1 and NSE was 75.6% (34/45), 53.3% (24/45), 66.7% (30/45) and 42.2% (19/45) respectively for lung cancer. The diagnostic sensitivity of IGF-1 combined with CYFRA21-1 was 95.5 %( 43/45) and that of IGF-1 combined with CEA and CYFRA21-1was 97.8%(44/45). Only IGF-1 and CYFRA21-1 showed significant changes before and after treatment (IGF-1: χ2=5.99,P=0.014; CYFRA21-1:χ2=4.99, P=0.025) in cancer group. Conclusions Serum IGF-1, CEA, CYFRA21-1 and NSE are all valuable for lung cancer diagnosis and the combination of those parameters can enhance the diagnostic efficiency. Serum IGF-1 and CYFRA21-1 may also be useful for evaluating the treatment response in lung cancer.

Correlation between adrenomedullin and VEGF in plasma of patients with lung cancer and the clinical significance / 国际肿瘤学杂志

Objective To investigate the expression of adrenomedullin (ADM) and vascular endothelial growth factor( VEGF) in plasm of patients with lung cancer,their correlation,and clinical significance. To explore the potential value of ADM for lung cancer in clinic. Methods The levels of plasms ADM and VECF were measured by means of radioimmunoassay in 66 cases of lung cancer, 31 cases of benign lung disease and 16 cases of healthy persons.Results The expression levels of plasm ADM and VEGF were significantly higher in the patients with lung caner than those in benign lung disease and healthy persons (P <0.05) , ADM in benign lung disease was significantly higher than that in the healthy persons (P <0.05), while VEGF in benign lung disease and healthy persons had no significant difference (P >0.05). The levels of plasm ADM and VEGF in distant metastasis was significantly higher than those with local lymph node metastasis ( P < 0.05 ) ,local lymph node metastasis was significantly higher than those without lymph node metastasis (P <0. 05). The levels of plasm ADM and VEGF were closely related to the local lymph node metastasis, distant metastasis and TNM staging. Furthermore, the expression levels of plasm ADM and VEGF increase with the clinical stage (P < 0.05 ). There was a significantly positive correlation between ADM and VEGF in patients with lung cancer (r = 0.449, P < 0.01) , but there was any correlation between benign lung disease group and healthy group ( P > 0. 05 ). Conclusion ADM and VEGF have synergistic effects in the process of neovascularization in lung cancer; to detect the expression of ADM, VEGF in plasm may be helpful for diagnosis of lung cancer and benign lung, lung cancer clinical staging,to interfere with ADM and its signal transduction pathway may be another new targeted therapy of lung cancer .

Clinical value of soluble intercellular adhesion molecule -1 in patients with thyroid disease / 中国基层医药

Objective To explore the contents and clinical significance of soluble intracellular adhesion molecule-1 ( sICAM-1 ) in patients with single goitre,Graves'and Hashimoto disease. Methods The contents of sICAM-1 in 100 cases of simple goiter group, Graves disease (GD) group 250 cases, Hashimot group 50 cases and 100 normal control were examined by sICAM-1 Radioimmunoassay(RIA) method,and the results were analyzed. Results There were no significant difference of sICAM-1 contents between ( 170.43 ± 34. 23 ) μg/L in normal control group and ( 182.48 ± 40.05) μg/L in simple goiter group( t = 1. 104, P ＞ 0. 05 ); The contents of slCAM-1 in GD group and HT group [( 279.93 ± 86.69) μg/L、 (250.36 ± 81.56) μg/L] were higher than the control group( t = 2.310,2. 210, all P ＜0. 05) ;The sICAM-1 contents in 3 species.methods after treatment [( 178.95 ±59.78) μg/L, ( 185.65 ±53.25)μg/L, (259.41 ± 71.46) μg/L)] were significantly lower than before treatment [(316.53 ± 66.13) μg/L, (277.79±64.30)μg/L,(285.71 ±72.14)μg/L](t=2.312,2.278,2.328,all P ＜0.05);After the Graves'patients were treated and their thyroid function were normal,their serum sICAM-1 levels( 251.92 ± 77.75 )μg/L were lower than that( 329.34 ± 90.47 ) μg/L in relapse Graves'group( t= 2.412 ,P ＜ 0. 05). Conclusion sICAM-1 RIA can be used as a parameter in diagnosing autoimmune thyroid diseases and in evaluating effects of therapy,stopping medicine or the relapse of Graves' disease.

Expression of endothelin-1 in cerebral vessels of scalded rats / 解剖学报

Objective To investigate the expression and effects of endothelin-1 (ET-1) in cerebral vessels of scalded rats. MethodHistological method was used to observe the alteration of histological structure of basilar artery in 8 scalded rats. Reverse transcription polymerase chain reaction(RT-PCR) was performed to examine expression of ET-1 mRNA;Western blotting to analyse expression of endothelin-1;radioimmunoassay to measure ET-1 levels;R3espectively, of cerebral basilar artery in 48 scalded rats. ResultVascular pathohistological changes were observed in cerebral basilar artery after rats were scalded;ET-1, ET-1 mRNA expression and ET-1 levels of basilar artery in scald 6 hours, 12 hours and 24 hours group were obviously higher than that of control group. Conclusion Scald may cause an increases of ET-1 expression in basilar artery, and elevated ET-1 expression may relate to the pathogenesis of cerebral vascular injury in scalded rats.