化学发光法测量人附睾蛋白4的不确定度的应用与分析

目的 探讨化学发光法测量人附睾蛋白4(HE4)不确定度的评定及不确定度在临床上的应用价值.方法 确定被测量物,分析不确定度来源,基于50.00、175.00 pmol/L两个室内质控水平合成、计算标准不确定度及扩展不确定度;基于以上两个水平的不确定度举例说明不确定度在临床上的应用.结果 不精密度、系统偏倚、校准物质不确定度为HE4测量不确定度的主要来源.50.00、175.00 pmol/L两个室内质控水平的标准不确定度分别为2.22,8.33,扩展不确定度分别为4.44,16.66;HE4不确定度可帮助临床医生判断同一患者前后两次测量结果差异是否具有显著性.结论 化学发光法测量HE4的不确定度主要来源为分析中的不精密度、系统偏倚及校准品的不确定度,分析前因素的不确定度评定较难,但可通过分析前质量控制加以控制,可不予考虑.不确定度的引入对临床医生诊断疾病、评估患者病情变化有重要意义.     

Protein gene product (PGP) 9.5 as a reliable marker in primitive neuroectodermal tumours—an immunohistochemical study of 21 childhood cases

A number of antibodies to neural proteins have been used to demonstrate neuronal differentiation in primitive neuroectodermal tumours. One of them is protein gene product (PGP) 9.5, a neuronal protein isolated from brain, whose function is unknown at present. We have studied differentiation in 21 cases of primitive neuroectodermal tumours of the CNS in children. Immunocytochemical staining was performed for such neuronal markers as: PGP 9.5, neuron specific enolase and synaptophysin, a glycosylated protein associated with synaptic vesicles. Positive staining for PGP 9.5 was present in 16 cases (strong staining in 12), for neuron-specific enolase in 16 cases (strong staining in 10) and for synaptophysin in 10 cases (strong staining in six). Both PGP 9.5 and synaptophysin showed a clear staining pattern with less non-specific background than with neuron-specific enolase. Our findings demonstrate the value of using more than one antibody marker in assessing neuronal differentiation in tumours. The high incidence of positive staining with antibody to PGP 9.5 suggests that this is an essential marker in the panel of antibodies used for the identification of primitive neuroectodermal tumours.     

HE4与CA125联检在卵巢恶性肿瘤诊断中的价值

目的:探讨人血清附睾蛋白4(HE4)和CA125在卵巢恶性肿瘤诊断中的价值.方法:检测卵巢恶性病变组(n=116)和良性病变组(n=92)女性患者血清HE4、CA125的水平;并与48例体检健康妇女(正常对照组)相比较.结果:在卵巢恶性病变组中,HE4、CA125的水平显著高于良性病变组和正常对照组(P＜0.05);H...     

Suppression by Human Placental Protein 14 of Natural Killer Cell Activity

ABSTRACT: Human decidua of early pregnancy contains considerable numbers of CD3^? CD56⁺ natural killer (NK) cells. In this study, two major protein products of the decidua, placental protein 14 (PP14) and placental protein 12 (PP12), were tested for the ability to regulate human NK cell activity. In vitro overnight exposure to PP14 of blood lymphocytes or purified large granular lymphocytes (LGL) resulted in suppression of cytotoxicity against K562 target cells in a 4-h ⁵¹Cr release assay. The NK inhibition was dependent on concentrations of PP14, being detectable at 5 μg/ml and reaching maximum at 50 μg/ml. Manifestation of PP14-induced NK suppression required 18-h contact with NK cells. The suppression of NK activity by PP14 was not abolished by indomethacin. In a target binding assay the number of PP14-treated LGL binding to K562 was comparable to that of untreated ones. By contrast with PP14, PP12 produced no effects on NK cells. These results indicate that PP14 suppresses the function of NK cells, which might be involved in prevention of maternal immune rejection of fetus at the fetomaternal interface.     

嗜酸性粒细胞阳离子蛋白在支气管哮喘中的研究

嗜酸性粒细胞阳离子蛋白(ECP)是活化嗜酸性细胞释放的强碱性蛋白中的重要组分,是嗜酸性粒细胞活化的重要标志,本文概述了ECP的来源,生物学作用以及与感染的关系,着重综述了ECP与支气管哮喘之间的关系。     

Endocrine differentiation of extra-pulmonary small cell carcinoma demonstrated by immunohistochemistry using antibodies to PGP 9.5, neuron-specific enolase and the C-flanking peptide of human pro-bombesin

Several recent studies have confirmed the endocrine nature of small cell carcinoma of the lung. In extra-pulmonary sites, small cell 'undifferentiated' carcinomas have classical morphological features similar to their pulmonary counterpart. We therefore investigated, using immunocytochemistry, the possibility that the non-pulmonary neoplasms may also be endocrine in nature. Sections of 29 small cell carcinomas from oesophagus, stomach, larynx, colon and urinary bladder were immunostained using antisera to protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), cytokeratin, leucocyte common antigen and peptides including bombesin, the C-flanking peptide of human probombesin, adrenocorticotrophic hormone, neurotensin, calcitonin and pancreatic polypeptide. All the tumours showed immunoreactivity for at least one of the two general endocrine markers PGP 9.5 and NSE. Twenty-three of the 29 cases were immunoreactive for PGP 9.5, 27 for NSE. All were positive for cytokeratin and negative for leucocyte common antigen. Of the regulatory peptides, immunoreactivity was obtained with antisera to bombesin (one case), the C-flanking peptide of human pro-bombesin (14 cases), adrenocorticotrophic hormone (one case) and calcitonin (three cases). No PGP 9.5-, NSE- or peptide-like immunoreactivity was detected in 25 control tumours from similar sites, including lymphomas and poorly differentiated tumours. These results suggest that non-pulmonary small cell carcinoma has an endocrine character.     

血尿酸、纤维蛋白原、超敏C-反应蛋白与急性脑梗死的关系

目的探讨血尿酸(Uric acid,UA)、血纤维蛋白原(Fibrinogen,Fg)、超敏C反应蛋白(high sensitive C-reactive protein,hs-CRP)在急性脑梗死中的意义。方法对脑梗死患者98例及对照组68例进行血UA、Fg、hs-CRP的检测。结果急性脑梗死患者血清UA(334.81±94.54)μmol/L、Fg(3.85±1.18)g/L、hs-CRP(5.08±5.52)mg/L水平明显增高,差异有显著性意义(P<0.05);UA与神经功能缺损程度无关,Fg、hs-CRP与神经功能缺损程度有关。结论Fg、hs-CRP与缺血性脑血管病的发生和神经功能缺损程度有关,UA与缺血性脑血管病的发生有关而与神经功能缺损程度无关。     

改进SP法检测热休克蛋白

目的 :探索避免常规SP法检测热休克蛋白 (HSP)时出现假阳性和消除非特异性反应的方法。方法 :心肌组织石蜡切片 ,采用改进的SP法染色 ,即在常规SP法基础上“一抗”倍比稀释 ,双蒸水稀释H2 O2阻断内源性过氧化物酶等染色操作。结果 :改进的SP法染HSP阳性产物呈棕黄色细颗粒状 ,在细胞浆中弥漫分布 ,背景十分清亮。结论 :改进后的SP法更适用于HSP的检测     

Immunohistochemical localisation of regulatory neuropeptides in human circumvallate papillae

The occurrence and distribution of neuropeptide-containing nerve fibres in the human circumvallate papillae were examined by the peroxidase–antiperoxidase immunolocalisation method using surgical specimens that had not been subjected to radiotherapy, and the abundance of neuropeptide-containing fibres was expressed as the percentage of total nerve fibres demonstrated by protein gene product (PGP) 9.5 immunoreactivity for a quantitative representation of these peptidergic fibres. Substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive (IR) nerve fibres were densely distributed in the connective tissue core of the circumvallate papillae, and some SP and CGRP-IR fibres were associated with the taste buds. A moderate number of vasoactive intestinal polypeptide (VIP)-IR fibres and a few galanin (GAL)-IR fibres were also seen in the connective tissue core and subepithelial layer. There were, however, no VIP-IR or GAL-IR fibres associated with the taste buds. Neuropeptide Y (NPY)-IR fibres were few and were associated with the blood vessels. Within the epithelium of the circumvallate papillae, no peptidergic fibres were found, although a number of PGP 9.5-IR fibres were detected. The abundance of SP, CGRP, VIP, and GAL-IR fibres expressed as the percentage of total PGP 9.5 IR fibres was 25.35±3.45%, 22.18±3.26%, 10.23±1.18%, and 4.12±1.05%, respectively. The percentage of NPY-IR fibres was below 3%. In a deeper layer of the papillae, a few VIP, GAL, and NPY-IR ganglion cells were found, and VIP immunoreactivity was detected in a few cells of the taste buds. There was no somatostatin, leucine enkephalin, or methionine enkephalin immunoreactivity in the circumvallate papillae. These results suggest that the dense SP and CGRP-IR fibres within the connective tissue core of the human circumvallate papillae may be involved in the deep sensation of the tongue.     

脑外伤患者血浆D-D、CRP和Fb的检测及临床观察

目的:揭示脑外伤患者血浆D-二聚体(D-D)、C-反应蛋白(CRP)和纤维蛋白原(Fb)浓度,分析其与预后的相关性。方法:收集脑外伤患者86例及对照组30例,检测血浆D-D、CRP和Fb浓度,统计分析其与预后的相关性。结果:脑外伤患者入院后1个月内死亡20例(23.3%)。脑外伤患者血浆D-D、CRP和Fb浓度较对照组明显升高(P<0.01),非生存患者血浆D-D、CRP和Fb浓度较生存患者明显升高(P<0.01),血浆D-D、CRP和Fb浓度预测脑外伤后1个月内死亡均有较高的灵敏度和特异度(P<0.01)。结论:D-D、CRP和Fb共同参与了脑外伤的病理生理过程,该些标志物适合作为一种常规的甄别方法,临床检测这些指标有助于早期预测脑外伤后1个月内死亡的评估。     

急性脑梗死患者血清Lp（a）、CRP、D-D、FG含量及临床意义

目的：探讨急性脑梗死与血中脂蛋白（a）[Lp（a）]以及C反应蛋白（CRP）、D-二聚体（D-D）和纤维蛋白原（FG）的关系。方法：在我院住院治疗的急性脑梗死患者335例,于入院次日清晨抽血检测其Lp（a）、CRP、D-D、FG的含量。同时选择性别和年龄相似的健康查体者作为对照组。比较不同梗死面积的急性脑梗死患者血液中Lp（a）、CRP、D-D、FG的水平。结果：①急性脑梗死组Lp（a）、CRP、D-D、FG的值分别为（425.13±85.11）mg/L、（16.32±5.21）mg/L、（285.11±116.33）ng/ml、（7.11±3.26）g/L,与对照组比较差异有显著性（P〈0.05或P〈0.01）。②不同梗死面积的急性脑梗死患者组间Lp（a）、CRP、D-D、FG的水平呈现大梗死灶〉小梗死灶〉腔隙性梗死的趋势,差异具有显著性（P〈0.05或P〈0.01）。结论：急性脑梗死的发生发展与血液中Lp（a）、CRP、D-D、FG的水平关系密切,Lp（a）、CRP、D-D、FG水平越高提示梗死面积越大,病情严重。Lp（a）、CRP、D-D、FG可作为急性脑梗死的危险因素,并可预测患者的梗死面积、病情严重程度。     

人GST-PTEN融合蛋白的原核表达和纯化

目的构建人第10号染色体上磷酸酶和张力蛋白同源缺失的基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)原核表达质粒,使其在原核细胞中高效表达并进行纯化。方法将全长片段插入原核表达载体pGEX-4T-1,构建重组子pGEX-4T-1-PTEN,转化BL-21感受态细胞。经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达PTEN融合蛋白的可诱导性表达,通过SDS-PAGE电泳、Western印迹分析证实蛋白表达的特异性。并用谷胱甘肽-S-转移酶(glutathione-Stransferase,GST)亲和层析对融合蛋白进行纯化。结果成功构建了原核表达载体pGEX-4T-1-PTEN,并将PTEN融合蛋白的成功表达,通过SDS-PAGE电泳、Western印迹分析,证实了蛋白表达的特异性。并对蛋白进行了纯化,获得了GST-PTEN融合蛋白的纯品。结论成功表达、纯化了GST-PTEN融合蛋白,为进一步研究PTEN蛋白的功能及其与其它功能性蛋白的相互作用研究奠定了基础。     

2型糖尿病患者超敏C反应蛋白与微量白蛋白尿的关系研究

目的探讨2型糖尿病患者超敏C反应蛋白（hs—CRP）的变化及微量白蛋白尿（MAU）与hs—CRP的关系。方法对160例2型糖尿病患者根据尿白蛋白排泄率（UAER）分为正常白蛋白尿组、微量白蛋白尿组，分别进行空腹血糖和hs—CRP测定，并与100例正常对照组比较。结果2型糖尿病患者hs—CRP明显高于正常对照组（P〈0．01），而微量白蛋白尿组hs—CRP值明显高于正常白蛋白尿组（P〈0．05）。结论hs—CRP与糖尿病及其血管并发症有关。微量白蛋白尿是心血管病的预测指标．慢性炎症可能起媒介作用。     

卵巢癌患者血清Hcy、HFA和SA联检的临床意义

目的：探讨了卵巢癌患者血清Hcy、HE4和sA联检的变化及临床意义。方法：应用酶联法和生化法对33例卵巢癌患者进行了血清Hcy、HFA和sA的检测,并与35例正常健康人作比较。结果：卵巢癌患者血清Hcy、HE4和SA水平均非常显著地高于正常人组（P〈0．01）。且血清Hcy水平与HE4、SA水平呈正相关（r=0．6128、0．5942,P〈0．01）。结论：检测卵巢癌患者血清Hcy、HFA和SA水平的变化,提示该类生物标志物的变化,有助于对疾病早期诊断和适宜的预后。     

癌症病人血清中免疫抑制酸性蛋白的测定

免疫抑制酸性蛋白（ＩＡＰ）是一种免疫抑制因子，在抑制带瘤宿主机体免疫监视系统中起着重要作用。本文介绍采用单向琼脂免疫扩散法，对多种肿瘤患者、非肿瘤患者及正常人血清ＩＡＰ水平进行检测。结果表明，肿瘤患者血清中的ＩＡＰ水平和异常率明显高于非肿瘤患者及正常人，提示ＩＡＰ可作为多种癌症的诊断、恶变程度以及术后观察、预后的临床参考指标。     

卵巢癌患者血清HE4、CA125和血浆LPA水平变化及其临床意义

目的:探讨血清HE4、CA125和血浆LPA水平在卵巢癌患者中的变化及其临床意义.方法:分别应用单克隆夹心ELISA、放射免疫分析和化学法定量检测50例卵巢癌患者血清HE4、CA125和血浆LPA含量,并与正常对照组比较分析.结果:卵巢癌Ⅲ/Ⅳ期患者血清CA125和血浆LPA水平均显著高于Ⅰ/Ⅱ期(P<0.01),而血...     

Immunocytochemical expression of protein gene product (PGP) 9.5 in the cat bronchopulmonary neuroendocrine cells and nerves

Variously fixed, wax-embedded lung and gastrointestinal serial tissue sections from newborn to adult cats were stained with hematoxylin-eosin (H&E), Grimelius' silver, and immunohistochemical techniques using antisera to protein gene product (PGP) 9.5, a neuron-specific protein under strong evolutionary constraints. PGP 9.5 is revealed as a pan-neuroendocrine marker useful for tracing the pulmonary diffuse neuroendocrine system (PDNES) and studying the relationships between neuronal and neuroendocrine elements at various stages of life. Its occurrence is also compared in the pulmonary and the gastrointestinal tract. In spite of a close resemblance to already described neuroepithelial bodies (NEB) of other mammals, cat NEB feature typical constitutional and distributional difference, illustrating interspecies differences. The number of PGP 9.5 immunopositive pulmonary neuroendocrine cells declines gradually after 3 weeks and throughout adult life. Immunoreactivity in neuronal elements is lost after 1 week of age. In gastrointestinal tissues, only neuronal lelements immunostain, suggesting functional variations or a separate embryological origin for enteroendocrine cells. © 1993 Wiley-Liss, Inc.