钙离子结合蛋白S100A9在乳腺癌中表达及临床意义

目的:探讨钙离子结合蛋白S100A9在乳腺癌患者中的表达及临床意义。方法:选取39例乳腺癌患者术前、15例术后血清及10例健康女性血清,12例手术切除肿瘤组织标本,应用ELISA检测乳腺癌患者血清S100A9水平,采用免疫组织化学方法检测乳腺癌组织中S100A9的表达,分析S100A9表达与临床病理特征的关系。结果:S100A9在乳腺癌组患者血清中表达水平高于健康组(P<0.01),有淋巴结转移乳腺癌组血清S100A9表达水平高于无淋巴结组(P<0.01),乳腺癌患者术后S100A9血清表达水平降低(P<0.01)。乳腺癌患者血清中S100A9水平与患者年龄、肿瘤最大径、组织学分级和病理类型有关(P<0.01)。S100A9在乳腺癌组织中表达明显高于癌旁正常组织。结论:S100A9表达水平在乳腺癌血清和组织中明显升高,表明S100A9可作为乳腺癌患者诊断和疗效观察指标。     

ELISA检测血清S100B蛋白方法的建立与临床应用

建立了S10 0B蛋白双抗体夹心ELISA检测方法 ,为临床提供快速、可信的定量指标。采用抗S10 0B蛋白单克隆抗体为包被抗体 ,抗S10 0多克隆抗体为检测抗体 ,对正常人和神经系统病变患者血清中S10 0B蛋白进行了检测。该法线性范围广 ,灵敏度高 ,重复性好。血清参考范围 (0 0 6 8～ 0 72 8) μg/L。 14 2份临床血清标本总阳性率为脑梗塞 2 1 6 %、脑挫裂伤33 3%、脑出血 5 9%。该法对临床脑及相关疾病的诊断及预后判断有很好的应用前景     

SARS冠状病毒N蛋白基因的表达及其在SARS诊断中的应用

目的 获得重组SARS冠状病毒（SARS-CoV）N蛋白抗原，建立特异性诊断SAILS病毒感染的免疫学方法。方法 大肠埃希菌中表达SARS病毒N蛋白基因，用金属螯合层析纯化N蛋白，建立检测SARS抗体的EUSA方法。结果 大肠埃希菌中表达了SARS病毒全长N蛋白抗原，经包涵体洗涤和金属螯和纯化后得到纯度较高的重组蛋白。用重组抗原EUSA检测30名SARS患者抗体全部为阳性，30名正常人血清为阴性，30名发热非SARS患者血清为阴性。结论 SARS表达核蛋白可以在大肠埃希菌中得到高效表达，纯化的重组N蛋白具有良好抗原性，可用于检测SARS抗体。     

S—100蛋白β亚单位ELISA检测方法的建立和初步应用

Ｓ１００蛋白是１９６５年由Ｍｏｏｒｅ发现并命名的一组酸性钙结合蛋白，相对分子质量（Ｍｒ）约为２００００，由α链和β链组成。根据组合方式，Ｓ１００可分为Ｓ１００ａ（αβ），Ｓ１００ｂ（ββ）和Ｓ１００ａｏ（αα）３种基本亚型。Ｓ１００的分布具有组织特异性，Ｓ１００ａ主要见于神经组织中的神经元，Ｓ１００ｂ主要见于胶质细胞中，Ｓ１００ａｏ则主要存在于心脏、横纹肌和肾脏等组织［１］。Ｓ１００作为病理学标志性蛋白，已用于黑色素瘤、神经胶质瘤、脊索瘤及脂肪瘤等的病理诊断。近期发现，体液中…     

出血性脑损伤患者脑脊液和血清中S100β蛋白的意义

目的:观察脑出血患者CSF和血清中S100β蛋白含量的变化及其临床意义。方法:以25例腹股沟疝和大隐静脉曲张患者作为25例脑出血患者对照组,取CSF和血清;将24只家兔随机分为两组(每组12只),一组实验组,另一组为假手术组。分别于实验性脑出血后6 h、12 h、24 h、48 h、72 h、96 h等各时间点取各组CSF和血清,采用ELISA法测定S100β蛋白的含量。结果:脑出血患者急性期与恢复期脑脊液中S100β蛋白的水平明显高于对照组(P<0.01);也明显高于血清S100β蛋白的水平(P<0.01);不同时间点兔脑出血模型实验组脑脊液中S100β蛋白的水平明显高于假手术组(P<0.01)。结论:脑出血后S100β蛋白在脑脊液中持续时间较长,且含量显著增高,可作为出血性脑损伤的早期诊断、指导治疗、判断预后的检测指标。     

SARS冠状病毒核衣壳蛋白的克隆表达及其临床应用研究

目的　克隆、原核表达严重急性呼吸综合征冠状病毒 (SARS-CoV)核衣壳 (N)蛋白 ,分析、评价其抗原性和在SARS血清学诊断中的应用价值。方法　采用逆转录巢式聚合酶链反应 (RT-nested PCR)扩增SARS CoV的N蛋白基因 ,克隆入pBAD-Thio TOPO原核表达载体 ,表达、纯化重组融合N蛋白 ,WesternBlot分析其抗原性和特异性 ,建立以重组N蛋白为抗原的酶联免疫吸附测定(ELISA)法 ,并与以全病毒裂解液为抗原的ELISA法进行比较。结果　重组表达载体经诱导产生了高水平的重组融合N蛋白 ,融合蛋白经亲和纯化后 ,具备了较高的纯度和抗原反应性 ,以重组蛋白为抗原的ELISA法在特异性和敏感性方面优于以全病毒裂解液为抗原者。结论　重组SARS-CoV的N蛋白具有良好的抗原性和特异性 ,可作为新一代SARS-CoV抗体检测试剂盒的备选抗原     

鼻咽癌组织中S100A4蛋白的表达及其临床意义

目的检测S100A4蛋白在鼻咽癌组织中的表达并探讨其与鼻咽癌的临床病理特征的关系。方法采用免疫组化SP法检测30例鼻咽慢性炎、122例鼻咽癌组织中S100A4的表达水平。结果 S100A4蛋白在鼻咽癌组织中的阳性率(76.2%)明显高于慢性炎组织(13.3%,P<0.01);N0～N1和N2～N3期中S100A4的阳性率分别为90.4%和65.7%,差异有统计学意义(P<0.01);临床分期中,Ⅲ～Ⅳ期和Ⅰ～Ⅱ期S100A4的阳性率分别为85.4%和51.5%,Ⅲ～Ⅳ期明显高于Ⅰ～Ⅱ期(P<0.01);此外,S100A4表达与患者性别、年龄、组织学类型以及T分期进展均无统计学意义(P>0.05)。结论 S100A4蛋白在鼻咽癌中表达升高,与淋巴结N分期和临床分期密切相关,有望成为判断鼻咽癌恶性生物学行为的有效指标。     

血清和精浆中C反应蛋白的ELISA测定

用抗人Ｃ反应蛋白（ＣＲＰ）血清和ＣＲＰ标准品建立了检测人血清和精浆中ＣＲＰ的ＥＬＩＳＡ，检测了正常人和不育男性精浆中ＣＲＰ含量，以及１０种疾病患者血清ＣＲＰ含量。结果表明，研制的试剂和建立的方法可满足临床常规检测要求。     

大鼠坐骨神经钳伤后变性和再生过程中S—100蛋白的变化

选用Ｗｉｓｔａｒ雌性大鼠４０只，分设正常，神经损伤，损伤对照组，运用免疫组化ＰＡＰ方法对神经干内的Ｓ－１００蛋白检测。染色结果经形态学观察及电子计算机图像处理系统分析结果表明：坐骨神经干内Ｓ－１００蛋白含量与损伤神经再生时间有密切关系，并观察到损伤第５，７天，损伤神经侧Ｓ－１００轴浆转运现象，推测Ｓ－１００可能参于了周围神经再生微环境的变化，从而影响损伤神经元的存活和神经纤维的再生。     

肠道病毒71型重组VP1蛋白的表达和EV71型手足口病血清学诊断方法的建立

目的 获得纯化的具有免疫活性的肠道病毒71型VP1蛋白,建立EV71感染早期、快速和准确的ELISA血清学诊断方法.方法 通过PCR方法扩增出VP1基因,定向克隆到原核表达载体pET-21b(+),阳性质粒转化入E.coli B121(DE3)感受态细胞经IPTG诱导,SDS-PAGE电泳和蛋白免疫印迹分析目的 蛋白的表达水平.纯化的VPI蛋白用作包被抗原,建立手足口病(HFMD)患者抗-EV71-IgM和IgG的血清学诊断方法.结果 成功表达和纯化了VP1重组蛋白,所表达的蛋白能被EV71型手足口病患者血清所识别.调查发现,与正常人和EV71阴性手足口病患儿比较,EV71阳性手足口病血清中抗-EV71-IgM和IgG中A值显著升高.差异具有统计学意义(P<0.05).与RT-PCR结果比较,发现该方法IgM的诊断敏感性和特异性分别为73%和77%;该IgG诊断方法的敏感性和特异性分别为82%和83%.完成该试验仪需4 h.结论 利用pET原核表达系统成功克隆、表达和纯化了肠道病毒71型重组外壳蛋白VPI,且具有良好的抗原性.该抗原可用于研制EV71血清学诊断试剂盒.     

S100A4蛋白在卵巢浆液性腺癌中的表达及其临床意义

目的: 分析S100A4蛋白在卵巢浆液性腺癌中的表达及其与临床病理因素和预后的关系,探讨其在卵巢浆液性腺癌侵袭转移过程中的作用及判断卵巢浆液性腺癌患者预后的价值。方法: 用免疫组化方法检测S100A4蛋白在卵巢浆液性腺癌、卵巢浆液性腺瘤及卵巢交界性浆液性腺瘤组织中的表达,分析S100A4蛋白的表达与卵巢浆液性腺癌各临床病理因素及生存预后的关系。结果: S100A4蛋白表达定位在肿瘤细胞的细胞质和细胞核,在卵巢浆液性腺癌、卵巢交界性浆液性腺瘤和卵巢浆液性腺瘤组织中的高表达率分别为78%、30%和27%,S100A4蛋白在卵巢浆液性腺癌组织中的高表达率高于其它两个对照组,差异有统计学意义。S100A4蛋白表达与卵巢浆液性腺癌患者的病理分级、治疗后是否复发有关(均P＜0.05)。单因素分析显示患者无疾病进展时间和总生存时间均与S100A4蛋白表达有关(P＜0.05)。多因素Cox回归分析显示术后残留病灶大小和病理分级是影响卵巢浆液性腺癌预后的独立因素。结论: S100A4蛋白表达上调在卵巢浆液性腺癌的发生过程中可能起一定作用。病理分级高的卵巢浆液性腺癌细胞可能部分通过上调S100A4蛋白表达增强其侵袭转移能力。S100A4蛋白可能成为对卵巢浆液性腺癌患者复发风险预测和预后判断的指标之一。     

Cerebrospinal fluid S100B levels reflect symptoms of depression in patients with non-inflammatory neurological disorders

Recent findings document numerous interactions between neuronal and glial systems that likely play a role in the pathophysiology of depression. These findings suggest that glia-derived neurotrophic protein S100B may play a significant role in developing depression. To test the relationship between S100B and depressive symptoms we designed cross-sectional clinical study including S100B serum and CSF levels in neurological patients with non-inflammatory disorders (NIND), who undergone cerebrospinal fluid assessment for diagnostic purposes. The present study was focused on psychometric testing of depression (BDI-II), anxiety (SAS) and alexithymia (TAS-20), and neurochemical measure of cerebrospinal fluid (CSF) and serum levels of S100B in 40 NIND inpatients [mean age 41.67]. The main result shows that S100B in CSF is significantly negatively correlated with BDI-II (Spearman R = −0.51, p < 0.0009) but not with SAS and TAS-20. The finding indicates that decreased level of S100B in CSF is related to increased symptoms of depression in the NIND patients.     

S100A4蛋白在甲状腺癌中的表达和意义

 目的 探讨S100A4蛋白在甲状腺癌组织中的表达及临床意义。方法 应用免疫组织化学SP法,检测56例甲状腺癌（乳头状腺癌30例,髓样癌12例,滤泡状癌8例,未分化癌6例）、45例甲状腺良性病变（甲状腺腺瘤、结节性甲状腺肿、原发性甲状腺功能亢进各15例）及13例甲状腺癌癌旁正常组织中S100A4蛋白的表达水平。结果 甲状腺癌组织中S100A4蛋白表达阳性率为83.9%（47/56）,表达强度呈（+）者25例,（++）者12例,（+++）者10例;甲状腺良性病变组织中S100A4蛋白表达阳性率为11.1%（5/45）,表达强度均为（+）;癌旁正常甲状腺组织中S100A4蛋白表达均呈阴性。甲状腺癌组织与后两者比较,差异均具有统计学意义（P<0.01）。临床分期Ⅰ～Ⅱ期者S100A4蛋白表达阳性率为77.8%（28/36）,表达强度为（+）者22例,（++）者4例,（+++）者2例;临床分期Ⅲ～Ⅴ期者表达阳性率为95.0%（19/20）,表达强度为（+）者3例、（++）者8例、（+++）者8例;两者间差异有统计学意义（P<0.01）。有淋巴结转移者S100A4蛋白表达阳性率为100%（16/16）,表达强度为（++）者8例、（+++）者8例;无淋巴结转移者表达阳性率为77.5%（31/40）,表达强度为（+）者25例、（++）者4例、（+++）者2例;两者间差异有统计学意义（P<0.01）。不同病理类型甲状腺癌组织中S100A4蛋白表达阳性率和表达强度差异无统计学意义（P>0.05）。结论 S100A4蛋白表达与甲状腺肿瘤细胞的恶性增殖和侵袭转移密切相关。S100A4蛋白表达强度作为判断甲状腺癌的恶性程度及预后的检测指标,具有临床应用的价值。     

Expression of S100 proteins in normal human tissues and common cancers using tissue microarrays: S100A6, S100A8, S100A9 and S100A11 are all overexpressed in common cancers

AIMS: To survey the expression of members of the S100 family of calcium-binding proteins in normal human tissues and common cancers using tissue microarrays. S100A6, S100A8, S100A9 and S100A11 have all been suggested to have potential roles in carcinogenesis and tumour progression but their expression has not been described in a wide range of human tissues and tumours. METHODS AND RESULTS: A custom-made tissue array, containing 291 tissue cores representing 28 tissue types and 21 tumour types, was used to produce sections that were immunostained for S100A2, S100A6, S100A8, S100A9, S100A11, calbindin 1, calbindin 2, S100B and parvalbumin. S100A6, S100A8 and S100A9 were expressed in 32%, 12% and 28% of breast cancers, respectively. There was a translocation of S100A11 expression from exclusively nuclear in normal tissues to cytoplasmic and nuclear in all common cancers. CONCLUSIONS: S100A6, S100A8, S100A9 and S100A11 are all expressed in common cancers, especially breast cancer. In addition, S100A11 undergoes a nucleocytoplasmic translocation which may have a direct influence on the proliferation of the cancer cells.     

S100A12在子痫前期患者母血中的表达及相关性研究

目的探讨S100A12在子痫前期患者母血清中的表达和临床意义。方法分别抽取24例子痫前期轻度患者、24例子痫前期重度患者以及24例正常妊娠妇女的血液,收集所有患者完整的病例资料,采用免疫印迹法检测血清中S100A12蛋白表达水平,并采双抗体夹心酶联免疫吸附法检测血清中TNF-α和IL-6的表达。结果与正常组相比,子痫前期患者血清中S100A12、IL-6和TNF-α的表达水平显著升高（P〈0.05）,且S100A12的表达水平与IL-6存在显著正相关性（P〈0.05）。结论 S100A12在子痫前期患者血清中的表达水平明显升高,有可能用于临床上子痫患者的诊断和治疗。     

川崎病急性期中性粒细胞功能及S100蛋白表达的变化

目的观察中性粒细胞在川崎病（KD）急性期的功能及S100A8/A9蛋白表达的变化,并探讨其意义。方法依据纳入标准和排除标准选取2006年11月至2007年7月在复旦大学附属儿科医院住院的KD患儿为研究对象,选取同期手术患儿为本研究对照组。通过二氢若丹明荧光染色法分析KD患儿中性粒细胞功能,并用荧光定量PCR法检测KD患儿中性粒细胞S100A8/A9 mRNA表达。结果共纳入KD患儿32例,其中男19例,女13例,年龄2个月至7岁2个月,平均（2.1±1.9）岁。冠状动脉损害者6例,无冠状动脉损害者26例;对照组手术患儿20例。KD急性期中性粒细胞明显活化,使用IVIG治疗后中性粒细胞活化百分比下降。KD急性期中性粒细胞S100A8/A9 mRNA的表达增加;IVIG治疗后,无冠状动脉损害患儿,S100A8/A9 mRNA表达明显降低,冠状动脉损害患儿S100A8/A9mRNA表达升高。结论KD急性期中性粒细胞活化,并存在相关的活化蛋白高表达,提示中性粒细胞可能参与了KD的发病机制。且在冠状动脉损害患儿中性粒细胞S100A8/A9 mRNA持续高表达,提示中性粒细胞可能参与冠状动脉损害。     

Accuracy of the determination of S100 protein expression in malignant melanoma using a polyclonal antibody directed against S100 and monoclonal antibodies specific for S100 alpha and beta

Abstract

S100 proteins are present in a variety of tissues and perform regulatory functions in numerous metabolic processes. They have an important role in many human cancers, including malignant melanoma. Both polyclonal and monoclonal antibodies have been used to investigate S100 expression in melanoma tissue sections. This study aimed to determine the accuracy and sensitivity of these two types of antibodies in detecting S100 proteins in paraffin processed tissue cases of malignant melanoma. The study compared routinely used rabbit polyclonal anti-S100 antibody raised against both anti-S100A and B isoforms (Dako, Glostrup, Denmark), as per studies by Timar, and compared and contrasted findings with mouse monoclonal anti-S100A and anti-S100B antibodies (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). The study involved the assessment of formalin-fixed paraffin-embedded tissue blocks from 56 cases of malignant melanoma, consisting of 23 superficial spreading, nine nodular, eight lentigo maligna, five acral lentigenous forms, five metastatic melanomas (two sentinel lymph node positive cases and three cases of nodal involvement from cases of elective nodal groin dissections), and six cases of desmoplastic malignant melanoma (DMM). The slides were stained by immunohistochemical methods on an automated platform (BenchMark XT; Roche, USA) and employing the iView detection system. All slides were examined by routine light microscopy by two independent assessors. The best results for both intensity of staining and percentage of positive tumor cells were achieved with polyclonal anti-S100 antibody and monoclonal anti-S100B antibody. Anti-S100A antibody yielded weaker staining intensity (with mean intensity of 1·8, compared to 2·8 for both anti-S100B antibody and polyclonal anti-S100 antibody), and a lower percentage of positive melanoma cells (an average of 74% for anti-S100A, compared to 95% for both anti-S100B antibody and polyclonal anti-S100 antibody). This result was statistically significant (P<0·01). Staining in cases of DMM gave the same results (P<0·01). The conclusion from this study is that polyclonal anti-S100 antibody and monoclonal anti-S100B antibody are more suitable than monoclonal anti-S100A antibody for diagnostic investigations of malignant melanoma, irrespective of the histological type of melanoma.     

Decrease of S100 beta protein in serum of patients with amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of unknown origin characterized by loss of upper and lower motor neurons and concomitant astrogliosis. We have investigated the S100 beta protein levels in serum as a marker for astroglia of patients with ALS (n=41) in comparison to a control group (n=32). Additionally we have investigated 12 patients at different follow-up time points (minimum 6 months). We could not observe a significant difference of S100 beta protein in patients with ALS in comparison to our control group (P=0.11) but we could clearly see a decrease of S100 beta levels in the further course of the disease. As S100 beta is also seen as a protein with nerve growth factor activity we assume that the fall of serum levels may reflect the loss of nerve growth stimulation in patients with ALS and suppose that repetitive measurements of S100 beta in serum can be used as an objective marker for disease progression.     

Evaluation of S100A4 mRNA in EUS-FNA specimens for the assessment of chemosensitivity to gemcitabine from patients with unresectable pancreatic cancer

Background/Aims: Gemcitabine (GEM) is the first-line chemotherapy in patients with unresectable pancreatic cancer. However, the clinical outcomes of this regimen are still unsatisfactory in prolonging survival. Resistant to GEM is one of the reasons for poor prognosis. Therefore, looking for molecular biomarkers to predict chemosensitivity to GEM is important for treatment in unresectable pancreatic cancer patients. The aim of this study was to analyze S100A4 mRNA in tissues of unresectable pancreatic cancer obtained by endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNA), and to determine the relation between S100A4 mRNA level and chemosensitivity to GEM. Methods: The analysis was performed on samples from 36 patients with unresectable pancreatic cancer who were treated with gemcitabine alone. The patients were assigned to receive GEM at 1,000 mg/m²/wk for weeks 1 to 6, followed by 1 week rest, then for 4 weeks. mRNA was extracted for S100A4 mRNA assay from patients above by EUS-FNA before GEM-treatment. The 36 patients were divided into the following two groups. Patients with partial response and those with stable disease whose tumor markers decreased by 50% or more were classified as the effective group. The rest of patients were classified as the non effective group. The relationship between GEM efficacy and S100A4 mRNA expression was then examined by chi-squared test. Results: S100A4 mRNA showed a significant correlation with GEM efficacy. Patients in the effective group had low S100A4 mRNA expression, whereas patients in non-effective group had high S100A4 mRNA expressions (P = 0.0059). Conclusion: S100A4 mRNA level analyzed in EUS-FNA samples is an important molecular biomarker for prediction of chemosensitivity to GEM in unresectable pancreatic cancer.