电化学发光免疫分析系统的性能评估

定量评估Elecsys2010电化学发光免疫分析系统的系统性能。以促甲状腺激素（Thyroid stimulating homromne,TSH)检测为例，对系统探测灵敏度，低，中，高质控血清的批内和批间精密度，批间稳定性参数Slope,ED20,ED50,ED80等4项指标，系统探测的准确度（回收试验），系统探测的有效性（平行试验）进行定量测定，TSH的最小测定值为0.004μU/ml。低，中，高3种质控样品批内和批间相对标准偏差（RSD）均小于5%，批间稳定性参数各RSD均小于5%，与统计学要求相一致，且符合标准曲线质控参数的重现性，回收试验满足临床要求，符合检验统计结果，理论值与实测值间作回归分析。相关系数r=0.999。回归方程表现良好的线性，且与标准曲线几乎完全重合，Elecsys2010电化学发光荧光免疫分析系统是一套有效性好，准确度高，灵敏度高，系统精度良好，检测速度快的自动化免疫分析系统。     

CA19-9时间分辨荧光免疫分析方法的建立及应用

以CA19－9单抗192＃包被板条,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合Eu^3 及标记241＃单抗,发光增强系统为以β-二酮体为主的增强液,采用平衡饱和法建立了CA19－9时间分辨荧光免疫分析。采用Log-Logit函数和四参数Logistic函数两种程序处理数据。结果表明方法的批内和批间CV分别显3．26％和5．74％,平均回收率为99．61％,灵敏度为1．66U/mL,可测范围为18．69－967U／mL,ED20、ED50t ED80分别为40．55U／mL、114．2U/mL和396．9U/mL。本方法与CEA有6％交叉反应,与CA125、CA153和AFP均无交叉反应,Eu^3 标记抗体于－30℃,保存六个月免疫反应性基本无损失,连续5个月应用同批试剂,分析结果稳定。78份血清样品的检测结果与化学发光吻合,本文提示,采用CA19－9时间分辨荧光免疫分析,可以高效灵敏地检测出血清中的CA19－9的含量,值得临床推广。     

C-肽化学发光免疫分析方法学研究

目的：建立人血清C-肽化学发光免疫分析法。方法：采用两株高特异的抗C-肽单克隆抗体，以一株包被微孔板制成固相抗体，另一株标记碱性磷酸酶，以金刚烷衍生物为底物，双抗体夹心法测定人血清C-肽浓度。结果：以5μg/ml单克隆抗体包被微孔板，酶结合物以1：5000稀释，在C-肽浓度0．1～15ng／ml范围内线性良好，线性方程为y=0．9575x-0．0869，相关系数0．99；灵敏度为0．01ng／ml，批内、批间变异系数分别为5．8％、8．4％；平均回收率98．4％，范围91．5％～105．0％。结论：文中建立的人血清C-肽化学发光免疫分析法，首次建立小分子人血清C-肽双抗体夹心化学发光免疫法测定。线性好，灵敏度较常规高出一个数量级，准确度和精密性高，能够满足一般临床诊断和科研的应用。     

降钙素原对儿童细菌性脑膜炎鉴别诊断的Meta分析

目的探讨降钙素原（PCT）对儿童细菌性脑膜炎的诊断价值。方法采用计算机检索获得PCT对儿童细菌性脑膜炎的诊断性研究文献,检索时间为建库至2013年7月,按照QUADAS标准评价纳入文献质量。采用Meta—Disc1．4软件进行Meta分析,对敏感度、特异度、阳性似然比、阴性似然比进行汇总和异质性检验,绘制综合受试者特征曲线（SROC）,行敏感性分析和异质性原因分析。使用Stata12．0软件判断发表偏移并绘制漏斗图。结果12篇文献进入Meta分析（中文5篇,英文7篇）。①汇总敏感度0．95（95％CI：0．92—0．97）,特异度0．89（95％CI：0．86—0．92）,阳性似然比11．09（95％CI：5．73～21．49）,阴性似然比0．07（95％CI：0．05～0．11）,诊断比值比122．01（95％CI：65．08—228．75）,SROC曲线下面积（AUC）0．9777,Q’指数0．933。②分别剔除样本量〈50文献、中文文献和回顾性研究文献的敏感性分析显示,上述诊断参数的95％CI与原数据有较大重叠。③特异度、阳性似然比的文献间存在显著的异质性,研究地域、PCT检测方法和细菌性脑膜炎诊断标准可能不是异质性产生的原因。④以PCT5肛g·L。界值诊断儿童细菌性脑膜炎的敏感度、特异度,SROCAUC和Q’指数最大。结论PCT在鉴别诊断儿童细菌性脑膜炎与病毒性脑膜炎中有较高的敏感度和特异度,但各研究间存在异质性,使用PCT鉴别诊断儿童细菌性脑膜炎仍需结合具体的临床情况。     

铁蛋白时间分辨荧光免疫分析法的建立

用夹心法建立铁蛋白(FER)时间分辨荧光免疫分析法(TRFIA),测定血清FER的含量。以FER单克隆抗体(McAb)806#包被板,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合Eu^3 及标记FERMcAb 803#,发光增强系统为以β-二酮体为主的增强液。采用双对数函数数据处理程序,方法的批内和批间CV分别为2．2％和3．7％,平均回收率为118％,灵敏度为0．5μg／L,可测范围为0．5-1000μg／L。ED20、ED50和ED80分别为27．8μg／L、88．5μg／L和247μg／L。Eu^3 FER McAb于-30℃保存至少4个月,免疫反应性基本无损失,同批试剂连续分析4个月结果稳定。本法与PE公司的FER-TRFIA试剂盒所得样品的测定值的相关系数为0．98。本文建立的FER-TRFIA法,是一种适用于人群贫血普查和各种肝脏疾病及肿瘤的联合诊断的良好方法,分析范围、灵敏度和稳定性均较理想,值得推广应用。     

降钙素原对新生儿败血症早期诊断价值的系统评价

目的系统评价降钙素原（PCT）对新生儿败血症早期诊断价值。方法用计算机检索CNKI、VIP等数据库。按两个纳入标准分别纳入有关PCT诊断新生儿败血症的研究,采用QUADAS工具进行质量评价,利用Metadisel．4计算各个研究的合并DOR,进行异质性分析,并计算敏感度、特异度、阳性似然比和阴性似然比,进行合并受试者诊断特征曲线（SROC）分析,获得SROC曲线下面积（AUC）。用Review Manager5软件进行其它诊断试验评价指标的Meta分析。结果按照标准一,纳入8个研究;按照标准二,纳入7个研究。8个研究间不存在异质性（x^2=3．28,P=0．858,12=0．0％）。8个研究合并敏感度为0．914,合并特异度为0．935,AUC为0．9783;7项研究间不存在异质性（x^2=3．24,P=0．779,I^2=0．0％）,合并敏感度为0．876,合并特异度为0．943,AUC为O．9578。结论从目前的研究看,PCT对新生儿败血症早期诊断有很高的价值,是很好的辅助诊断指标。     

应用化学发光免疫分析法测定血清抗TgAb和抗TPOAb的临床诊断价值

为探讨化学发光免疫分析法 (CLIA)测定血清抗甲状腺球蛋白抗体 (抗TgAb)和抗甲状腺过氧化物酶抗体 (抗TPOAb)的临床诊断价值 ,以病理诊断作为主要依据 ,2 89例各种甲状腺疾病患者为研究对象 ,采用CLIA法测定血清抗TgAb、抗TPOAb水平 ,应用ROC曲线分析法评价CLIA法的诊断价值并确定诊断临界值。结果发现对自身免疫性甲状腺疾病 (AITD)的诊断 ,血清抗TgAb和抗TPOAb水平的ROC曲线下面积分别为 0 792± 0 0 2 9和 0 892±0 0 2 2 ,其诊断临界值分别为 90U mL(准确度 75 1% ,敏感度 6 3 3% ,特异度 82 2 % )和 30 0U mL(准确度 85 5 % ,敏感度 78 9% ,特异度 89 4 % )。特别针对桥本病的诊断 ,血清抗TgAb和抗TPOAb曲线下面积分别为 0 76 9± 0 0 4 4和 0 737± 0 0 4 9,诊断临界值分别为 30 0U mL(准确度 6 9 7% ,敏感度 5 4 9% ,特异度 82 8% )和 30 0 0U mL(准确度74 3% ,敏感度 70 6 % ,特异度 77 6 % ) ,总预测正确率为 71 6 % ,总判别功效为 6 9 7% ,增加血清TSH水平变量后 ,总预测正确率和总判别效力均提高至 89 9%。表明CLIA法测定血清抗TgAb和抗TPOAb水平对AITD诊断具有较高的诊断价值 ;对桥本病和Graves病的诊断 ,如同时参考血清促甲状腺激素 (TSH)水平 ,可明显提高诊断的准确度。     

甲状腺激素放射免疫分析内质控的实验与思考

目的 :探讨甲状腺激素T3 、T4、FT3 、FT4、TSH检测结果不稳定和各项目间的不匹配原因 ,以提高检测质量 ,为临床提供可信的报告。方法 :通过实验室内质控改进分离方法 ,对各类加样器及放免测定仪定期监测 ,放免指标采用零管结合率 (B0 )、非特异结合 (NSB)、有效剂量值 (ED2 0 、ED50 、ED80 )、平均批变异系数 (ABCV)、平均病人值和质控血清含量的Shewart质控图 ,免放法选用B0 、NSB、ED80 、ED50 、ED2 0 、标准点偏差、质控血清和平均病人值。结果 :通过随意选择 1 0批标本共 96 4例测定结果分析 ,有 1 5 4例项目之间不匹配占 1 6 % ,而方法和操作的误差仅占不匹配标本的 5 8% ,其中大多数为甲状腺激素的生化特性和临床表现。结论 :分离误差是造成随机误差最主要来源 ,应选择稳定性好的分离法 ,推进固相分离替代均相分离 ,平均病人值有补充质控血清的作用 ,它和内质控的各项参数组成实验结果偏移取舍指标 ,以去除因方法和操作引起的误差 ,余下的不匹配结果原因为临床甲状腺疾病特点。实验人员要从技术和临床角度来分析结果 ,沟通实验室与临床的联系 ,才能知己知彼地提供给临床可信的报告     

时间分辨荧光免疫法测定血清AFP的方法学评价

对时间分辨荧光免疫法(TRFIA)测定血清AFP的方法学予以评价.评价TRFIA法测定AFP的线性范围、灵敏度、精密度、回收率及与ELISA法和RIA法的相关性.结果显示:AFP线性范围1～1000U/mL,检测下限≤0.1 U/mL,批内及批间变异系数分别为2.3%、5.3%,回收率97.85%,与ELISA法及RIA法高度相关,r值分别为0.982与0.991.正常血清AFP参考值0.8～10.5 U/mL.结论:TRFIA测定AFP线性范围宽,稳定性好,灵敏度与准确度高,无放射性污染,是当前免疫分析中有发展前途的一项超微量检测技术.     

胃蛋白酶原Ⅰ时间分辨荧光免疫分析法的建立

目的　采用时间分辨荧光免疫分析技术建立高灵敏的胃蛋白酶原Ⅰ的快速全自动检测方法。方法　以PGⅠ单克隆抗体 80 0 3#包被板 ,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合Eu3 及标记PGⅠ单克隆抗体 80 16 # ,发光增强系统为以 β 二酮体为主的增强液。采用平衡饱和法建立PGⅠ时间分辨荧光免疫分析 ,数据采用Log Logit法函数和四参数Logitc函数数据处理程序处理。结果　方法的批内和批间CV分别为 1.9%和 4 .7% ,平均回收率为 10 2 .6 5 % ,灵敏度为 0 .0 5 μg L ,可测范围为 3.5～ 32 8μg L ,ED2 0 、ED50 和ED80 分别为 11.34μg L、38.73μg L和 132 .3μg L。Eu3 标记抗体 2 0℃保存 8个月免疫反应性基本无损失 ,同批试剂连续 8个月应用分析结果稳定 ,临床结果与免疫放射分析法的结果相符。结论　胃蛋白酶原Ⅰ时间分辨荧光免疫分析法是目前胃蛋白酶原Ⅰ检测中最灵敏的方法 ,该分析法稳定性好 ,具有很好的应用前景。     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.     

The development of the ligament of the head of the femur

The hip joints of 30 human male and female fetuses and stillborns between 20 mm and 350 mm crown-rump length were studied by light microscopy. The ligament of the head of the femur developed in situ as a condensation of mesenchyme at the end of the second month of intra-uterine life (IUL), and was vascularized by branches of acetabular vessels early in the fourth month. In the majority of fetuses older than 5.5 months IUL, vessels in the ligament passed a short way into the femoral head within cartilage canals, to supply a small region around the fovea capitis. The remainder of the head was supplied by vessels in canals from around the upper part of the neck. The ligament changed from predominantly cellular to fibrous during the last 4 months of IUL. This increase in strength suggested significant mechanical functions in utero: limitation of adduction-flexion and opposition to postero-superior dislocation were the most likely.     

Counterpointing the functional role of the forebrain and of the brainstem in the control of the sleep-waking system

This paper reviews the lifetime contributions of the author to the field of sleep-wakefulness (S-W), reinterprets results of the early studies, and suggests new conclusions and perspectives. Long-term cats with mesencephalic transection show behavioral/polygraphic rapid eye movement sleep (REMS), including the typical oculo-pupillary behavior, even when the section is performed in kittens prior to S-W maturation. REMS can be induced as a reflex. Typical non-rapid eye movement S (NREMS) is absent and full W/arousal is present only after a precollicular section. The isolated forebrain (IF) rostral to the transection exhibits all features of W/arousal and NREMS [with electroencephalographic (EEG) spindles and delta waves], arousal to olfactory stimuli, and including the appropriate oculo-pupillary behaviors. These features also mature normally after neonatal transection. REMS is absent from the IF. After deprivation there is NREMS pressure and rebound in the IF, but the decerebrate cat only shows pressure for REMS. Most IF reactions to pharmacologic agents are within expectations, except for the tolerance/withdrawal effects of long-term morphine use which are absent. In contrast, these effects are supported by the brainstem (i.e. seen in the decerebrate cat). In cats with ablation of the telencephalon, or diencephalic cats, delta waves are absent in the thalamus. EEG thalamic spindle waves are seen triggering S for only 4-5 days after ablation. Therefore, true NREMS is absent in chronic diencephalic cats although pre- and postsomniac behaviors persist. These animals are hyperactive and show a pronounced, permanent insomnia; however, a low dose of barbiturate triggers a dramatic REMS/atypical NREMS rebound. Cats without the thalamus (athalamic cats), initially show a dissociation between behavioral hyperactivity/insomnia and the neocortical EEG, which for 15-20 days exhibits only delta and slower oscillations. Fast, low-voltage W rhythms appear later on, first during REMS, but spindle waves and S postures are absent from the start, such that these cats also display only atypical NREMS. Athalamic cats also show barbiturate-sensitive insomnia. Cats with ablation of the frontal cortices or the caudate nuclei remain permanently hyperactive. They also show a mild, but significant hyposomnia, which is permanent in afrontal cats, but lasts for about a month in acaudates. The polygraphic/behavioral features of their S-W states remain normal. We conclude and propose that: (a) the control of the S-W system is highly complex and distributed, but is organized hierarchically in a well-defined rostro-caudal manner; the rostral-most or highest level (telencephalon), is the most functionally complex/adaptative and regulates the lower levels; the diencephalic/basal forebrain, or middle level, has a pivotal role in inducing switching between S and W and in coordinating the lowest (brainstem) and highest levels; (b) W can occur independently in both the forebrain and brainstem, but true NREMS- and REMS-generating mechanisms exist exclusively in the forebrain and brainstem, respectively; (c) forebrain and brainstem S-W processes can operate independently from each other and are preprogrammed at birth; this helps understanding normal and abnormal polygraphic/behavioral dissociations in humans and normal dissociations/splitting in aquatic mammals; (d) NREMS homeostasis is present in the IF, but only REMS pressure after deprivation persists in the decerebrate cat; (e) the thalamus engages in both NREMS and W; (f) insomnia in diencephalic cats is the result of an imbalance between antagonistic W- and S-promoting cellular groups in the ventral brain (normally modulated by the telencephalon); (g) the EEG waves, which are signature for each S-W state, appear to truly drive the concomitant behaviors, e.g. a hypothetical human IF could alternate between behavioral NREMS and W/arousal/awareness; (h) a role for REMS is to keep the individual sleeping at the end of the self-limiting NREMS periods. The need for accelerating research on telencephaling NREMS periods. The need for accelerating research on telencephalic S-W processes and downstream control of the lower S-W system levels is emphasized.     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Involvement of the striatum in the central regulation of the hormonal functions of the gonads

Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms. Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 85, No. 4, pp. 594–597, April, 1999.     

Ultrastructure of the endothelium of the drainage system of the eye

The endothelium of the ocular drainage system (Schlemm’s canal, collector tubules, and aqueous veins) in primary juvenile glaucoma undergoes degenerative dystrophic changes with compensatory hypertrophy and proliferation at the initial stages of the glaucomatous process and atrophy and desquamation at advanced and terminal stages. Progressive decrease in the pinocytous function of endotheliocytes, reduction of the protein-synthesizing and mitochondrial compartments of the cytoplasm, and formation of autophagosomes reflect the process of endotheliocyte degeneration in general. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 574–577, May, 2008