链球菌蛋白G应用于ELISA的实验研究

对酶标记链球菌蛋白Ｇ（ＳＰＧ）在ＥＬＩＳＡ的应用条件进行了研究并与ＳＰＡ作了系统比较。ＳＰＧ与ＩｇＧ结合受温度影响，３７℃结合活性高于室温和４℃；反应时间定为２ｈ反应溶液ｐＨ值升高，ＳＰＧ的结合力亦增强。在同等条件下与酶标记ＳＰＡ进行比较，结果表明ＳＰＧ对小鼠单克隆抗体、羊抗人、兔抗ＩＦＮ－γ和驴抗兔Ｉｇｇ以及牛、大鼠和人ＩｇＧ的结合力均明显高于ＳＰＡ。ＳＰＧ用于大鼠血清抗ＩＦＮ－γ抗体的检测，其阳性率和抗体滴度均大大高于ＳＰＡ。因此，ＳＰＧ作为一种广谱的、高亲和力的ＩｇＧ结合物，是应用于ＥＬＩＳＡ的理想的替代二抗。     

Scintillation proximity assay in lead discovery

Background: Scintillation proximity assay (SPA) is a homogeneous scintillant bead-based platform for the measurement of biological processes and plays an important role in the identification of active chemical entities in drug discovery. Objective: The design and development of solid-phase SPA approaches are examined and compared with alternative non-radiometric fluorescence-based technologies. Methods: This review provides background on the principle of SPA and its application to biomolecular interactions from a variety of biological sources. Conclusion: The SPA approach is well suited to the demands of commercial high volume automation and assay miniaturization for target-based high-throughput screening campaigns on synthetic and natural product libraries as well as for benchtop characterization and confirmation studies. In the near future, innovations in the way SPA and fluorescence-based screening strategies are multiplexed will improve our comprehensive understanding of cellular system biology and dramatically advance the lead discovery process for the treatment of complex target-related disorders.     

Orexin A decreases lipid peroxidation and apoptosis in a novel hypothalamic cell model

Current data support the idea that hypothalamic neuropeptide orexin A (OxA; hypocretin 1) mediates resistance to high fat diet-induced obesity. We previously demonstrated that OxA elevates spontaneous physical activity (SPA), that rodents with high SPA have higher endogenous orexin sensitivity, and that OxA-induced SPA contributes to obesity resistance in rodents. Recent reports show that OxA can confer neuroprotection against ischemic damage, and may decrease lipid peroxidation. This is noteworthy as independent lines of evidence indicate that diets high in saturated fats can decrease SPA, increase hypothalamic apoptosis, and lead to obesity. Together data suggest OxA may protect against obesity both by inducing SPA and by modulation of anti-apoptotic mechanisms. While OxA effects on SPA are well characterized, little is known about the short- and long-term effects of hypothalamic OxA signaling on intracellular neuronal metabolic status, or the physiological relevance of such signaling to SPA. To address this issue, we evaluated the neuroprotective effects of OxA in a novel immortalized primary embryonic rat hypothalamic cell line. We demonstrate for the first time that OxA increases cell viability during hydrogen peroxide challenge, decreases hydrogen peroxide-induced lipid peroxidative stress, and decreases caspase 3/7 induced apoptosis in an in vitro hypothalamic model. Our data support the hypothesis that OxA may promote obesity resistance both by increasing SPA, and by influencing survival of OxA-responsive hypothalamic neurons. Further identification of the individual mediators of the anti-apoptotic and peroxidative effects of OxA on target neurons could lead to therapies designed to maintain elevated SPA and increase obesity resistance.     

SPA—ELISA一步法检测囊虫病患者血清特异性抗体的研究

用ＳＰＡ－ＥＬＩＳＡ一步法检测了囊虫患者血清特异性抗体，并与快速ＳＡＰ－ＥＬＩＳＡ进行对比。两法同时检测了５９例囊虫病患者，３６例囊虫病病人和３０例正常人血清。结果表明：ＳＡＰ－ＥＬＩＳＡ一步法和快速ＳＰＡ－ＥＬＩＳＡ的阳性率分别为９４．９１％和９８．３１％，几何平均滴度分别为１：４２５．８９和１：５６５．７２，均无显著性差异。对１例肝吸虫病和１例包虫病人血清均起交叉反应。说明ＳＰＡ－ＥＬＩＳＡ一     

西宁地区健康人骨矿物含量调查分析

目的了解西宁地区健康人各年龄段骨矿物含量正常值。方法用SD-1000单光子骨矿物测定仪窄束γ-线通过非优势侧桡、尺骨中下1/3交界处测试点横向扫描。获得骨线密度(BMC)、面密度(BMD)及桡、尺骨骨密度均值。结果1.男性的骨密度在31~40岁为峰顶,女性在41~50岁达峰;2.性成熟期(18~44岁)骨密度值最高,此后随增龄而下降;3.体重与骨矿的含量相关;4.按WHO制定的低于同性别性成熟期骨密度2.5 s为骨质疏松的诊断标准,女性老年期为此症者占24.2%,男性老年期为4.60%,女性为男性5.3倍;5.按低于同性别性成熟期骨密度1 s为低骨量,男、女老年期分别为54%、82.3%。结论1.女性骨峰值较男性晚10年;2.男女性者骨密度值在性成熟期(18~44岁)达峰,此后骨矿物质含量丢失随年龄增长增多;3.体胖者骨矿含量相对较多;4.老年期骨质疏松发生率女性比男性高5.3倍,表明性腺的衰退时间对骨矿物的丢失有重大影响;5.西宁地区低骨量的高危人群众多,应该行早期行干预治疗。*     

一种基于集对分析的中西医结合模型研究

从系统和数学的层面上研究中西医结合的同、异、反现象及其内在本质,具体地采用集对分析的同异反系统理论和联系数,就中西医结合的同、异、反建立同异反系统学模型,借助模型来探索中西医结合中的同异反时空展开,在同异反时空展开过程中,探索中西医结合的实质,并用于中西医结合临床实践和疗效评定,同时也对中医学传统的“同病异治”“异病同治”“通因通用”“相反相成”等治则治法提供现代科学理论依据。     

E-coli和Yeast菌体蛋白ELISA定量测定方法的建立

目的:提高酶联免疫吸附测定(ELISA)方法的灵敏度。方法:将聚苯乙烯微孔板(PS)经过紫外线(UV)辐照后,用葡萄球菌A蛋白(SPA)预包被,再应用生物素-链霉亲和素(BSA)反应建立双抗体夹心ELISA。结果:检测低限为1.95ng/ml,方法的批内变异分别为7.96%和8.63%。结论:为菌体蛋白的测定提供了一种新的方法。     

Activity of growth factors in the IL-6 group in the differentiation of human lung adenocarcinoma

The role of the interleukin-6 (IL-6) group of cytokines in differentiation of two lung adenocarcinoma cell lines has been examined using induction of alkaline phosphatase and expression of surfactant protein A. Oncostatin M was the most active and potent for alkaline phosphatase in A549 cells, with IL-6 having similar activity but less potency. Neither cytokine induced alkaline phosphatase in NCI-H441 cells, although induction was obtained with lung fibroblast-conditioned medium. Surfactant protein A was induced in NCI-H441 cells by conditioned medium and dexamethasone and, to a much lesser extent, by oncostatin M or IL-6. Induction of alkaline phosphatase and surfactant protein A were both dexamethasone-dependent, though some induction of surfactant protein A was obtained with interferon-alpha in the absence of dexamethasone. The activity present in lung fibroblast-conditioned medium suggests paracrine control, but this appears not to be due to oncostatin M or IL-6 as disabling antibodies to either cytokine were not inhibitory, and, although alkaline phosphatase was induced in A549 by both cytokines, it was only induced by conditioned medium in NCI-H441 cells. Furthermore, surfactant protein A was induced in H441 by conditioned medium to a much greater extent than by oncostatin M or IL-6. These data demonstrate that cytokines of the IL-6 group have potential as differentiation inducers in lung adenocarcinoma cells and that there is an equivalent paracrine factor(s) in lung fibroblast conditioned medium. As the production of this factor by fibroblasts is not enhanced by glucocorticoid, although the response of the target cell is, it would appear to be distinct from the fibrocyte pneumocyte factor previously described by Post et al 1984.     

Morphology and the sperm penetration assay

OBJECTIVE: To assess the ability of Kruger morphology to predict the outcome of the sperm penetration assay (SPA). DESIGN: A retrospective review using univariate and multivariate analysis, t tests, and logistic regression was performed to examine the correlation between Kruger morphology and the optimized zona-free hamster egg sperm penetration assay (O-HESPA). SETTING: University hospital. PATIENT(S): One hundred fifteen private-practice patients who underwent semen analysis, including Kruger morphology and O-HESPA, as part of an infertility evaluation between 1997-2000 were retrospectively reviewed. INTERVENTION(S): Retrospective analysis of the sperm penetration assay. MAIN OUTCOME MEASURE(S): Univariate and multivariate analysis, Student's t test, and logistic regression were performed to analyze Kruger morphology, count, and viability and their relationship to SCI result. RESULT(S): Univariate analysis demonstrated a statistically significant correlation between SCI and sperm count, viability, and Kruger morphology. Multivariate analysis demonstrated statistically significant correlations between SCI and count and viability. There was no correlation between Kruger morphology and SCI. Logistic regression was performed on the SCI results, using count, Kruger morphology, and viability. Sperm count was found to be the only variable that was statistically significant with respect to SCI results. CONCLUSION: This study demonstrated that Kruger morphology assessment cannot be used to predict the results of SCI or replace it in the management of the infertile couple.     

Standardized phase angle indicates nutritional status in hospitalized preoperative patients

There is currently no criterion standard to assess nutritional status, and different methods have been used in hospitalized patients. The aim of this study was to investigate the agreement and the association between bioelectrical impedance analysis derived by standardized phase angle (SPA) and other methods used for the nutritional assessment of body composition, metabolic status, and functional status in surgical patients. The hypothesis was that the SPA is effective for evaluating nutritional status in surgical patients; therefore, it could be used when the application of other assessment methods is not possible. The sample consisted of 125 patients (20-94 years of age) before elective gastrointestinal or hernia repair. The participants were from the Surgical Clinic 1 at the University Hospital of the Federal University of Santa Catarina, Florianópolis, SC, Brazil. Nutritional status was evaluated during the preoperative period based on the triceps skinfold thickness, mid-arm circumference, body mass index, percent weight loss, nutritional risk screening 2002 (NRS 2002), subjective global assessment (SGA), and SPA. The agreement between the SPA and the other methods was assessed with the k coefficient. The agreement between the SPA and the methods of nutritional assessment that were investigated for diagnosing malnutrition was moderate for NRS 2002 and SGA, weak for percent weight loss, and poor for triceps skinfold thickness, mid-arm circumference, and body mass index. There was a significant association between SPA and both NRS 2002 and SGA (P < .001). Our results suggest that SPA is able to indicate the risk of nutritional deficiency in the patients assessed. However, good agreement between SPA and the methods investigated was not observed.