Nucleophosmin/B23在肝细胞癌组织中的高表达

目的探讨Nucleophosmin／B23（B23）在肝细胞癌（HCC）组织中的表达及其临床病理意义。方法采用重组蛋白表达和杂交瘤细胞技术制备B23重组蛋白和鼠抗B23单克隆抗体。收集103例HCC组织、12例肝局灶性结节性增生和17例肝血管瘤旁肝组织的临床病理档案资料,10例HCC及癌旁肝新鲜组织,采用免疫组织化学（ABC）方法、逆转录聚合酶链反应（RT-PCR）及免疫印迹等技术检测B23在这些组织中的表达,并与增殖细胞核抗原（PCNA）的表达相比较。应用统计学方法对结果进行分析。结果RT-PCR及免疫印迹结果显示：在mRNA和蛋白质水平,B23在HCC组织中的表达明显高于对应的癌旁肝组织。免疫组织化学结果显示：B23在4组（HCC组、癌旁肝组织组、肝局灶性结节性增生组、血管瘤旁肝组织组）中的表达差异有统计学意义（P〈0．001）,其中B23在HCC组织中的表达显著高于其他3组（P〈0．01）,PCNA在HCC组织中的表达也显著高于其他3组（P〈0．01）,相关性分析表明B23与PCNA在4组中表达强度及其差异具有相关性（r=0．4767,P〈0．01）。B23在HCC组织中的表达强度与患者血清AFP水平、肿瘤病理分级及是否伴有肝硬化之间的关系有统计学意义（P〈0．05）。结论B23在HCC中呈高表达,且显著高于非癌肝组织;B23可作为HCC细胞增生程度的潜在标记,并在临床病理上具有潜在的应用意义。     

Expression of gLTP in Sympathetic Ganglia from Stress-hypertensive Rats: Molecular Evidence

We previously reported behavioral and electrophysiological evidence indicating that superior cervical ganglia (SCG) from rats that developed hypertension as a result of chronic psychosocial stress expressed ganglionic long-term potentiation (gLTP) in vivo. In the present study, we present additional supportive evidence by measuring changes in protein levels of essential signaling molecules in ganglia from chronically stressed rats. We compared protein levels of essential, LTP-related signaling molecules in ganglia isolated from chronic stress-hypertensive rats, known to have expressed gLTP, with those of the same molecules in normal ganglia 1h after eliciting gLTP by high frequency stimulation (HFS) in vitro. Immunoblot analysis showed a significant increase in the levels of phosphorylated CaMKII, total CaMKII, nitric oxide synthase (NOS-1), and calmodulin in SCG from both chronically stressed rats and from normal rat ganglia in which gLTP was expressed by HFS in vitro. Additionally, there was a parallel reduction in calcineurin protein levels in ganglia from both groups. The present results confirm that ganglia from stressed rats have expressed gLTP in vivo and that synaptic plasticity in sympathetic ganglia may involve a molecular cascade largely similar to that of LTP in the hippocampal CA1 region.     

Evaluation of the role of H pylori infection in pathogenesis of gastric cancer by immunoblot assay

INTRODUCTION H pylori, a Gram-negative bacterium, is now widely considered as one of the major etiologic factors in the pathogenesis of a great variety of gastrointestinal diseases such as gastritis, peptic ulcers and mucosa- associated lymphoid tissue ly…     

Anti-U1RNP antibodies in patients with localized scleroderma

Abstract Antibodies to U1 ribonucleoproteins (RNP) have been detected in serum from patients with various autoimmune diseases. However, the presence of anti-U1RNP antibodies in patients with localized scleroderma has not been reported. In this study, we examined the frequency of anti-U1RNP antibodies using immunoprecipitation of U small nuclear RNAs and determined the antigen specificity by immunoblotting. Of 70 serum samples from patients with localized scleroderma, 2 (3%) immunoprecipitated U1 small nuclear RNA. Indirect immunofluorescence using HEp-2 cells as substrate showed coarse speckled nuclear fluorescence without nucleolar staining in both of the samples positive for anti-U1RNP antibodies. In addition, the presence of anti-U1RNP antibodies in each serum sample was confirmed by immunodiffusion against HeLa cell extracts. Immunoblotting analysis showed anti-70 kDa antibodies in each serum sample. This reaction against 70 kDa protein in the patients with localized scleroderma was analogous to that in patients with systemic sclerosis or mixed connective tissue disease. Both patients with positive serum were diagnosed as having linear scleroderma, but neither had evidence of Raynaud’s phenomenon or sclerodactyly. These results indicate that the presence of anti-U1RNP antibodies is one of the serological abnormalities in localized scleroderma, and that the mechanism of induction of anti-U1RNP antibodies in patients with localized scleroderma might be similar to that in patients with systemic sclerosis and mixed connective tissue disease. Received: 12 February 2001 / Revised: 27 April 2001 / Accepted: 11 July 2001     

抗Scl-70和抗Jo-1抗体在系统性硬皮病和皮肌炎中的临床意义

为了提高对系统性硬皮病及皮肌炎的临床确诊率，通过ＥＮＡ抗原应用免疫印迹技术检测了１９例系统性硬皮病（ＳＳ）及２８例皮肌炎或多发性肌炎（ＤＭ／ＰＭ）患者抗Ｓｃｌ ７０及抗Ｊｏ １抗体，并与系统性红斑狼疮（ＳＬＥ）、混合性结缔组织病（ＭＣＴＤ）共６２例及５０例健康人作了对照研究。结果表明：７０ ＫＤ（抗Ｓｃｌ ７０）多肽抗体是系统性硬皮病的特异性标记抗体，阳性率３１．６％；５５ＫＤ（抗Ｊｏ １）多肽抗体是皮肌炎的特异性标记抗体，阳性率４６．４％。ＳＬＥ、ＭＣＴＤ及健康人全部阴性，从而有助于对系统性硬皮病及皮肌炎的鉴别诊断     

NMDA-receptors 1 and 2A/B coassembly increased in human epileptic focal cortical dysplasia

PURPOSE: This study was designed to quantify the relation between expressions of NMDA receptor (NMDAR) subunits (1 and 2A/B) and the epileptogenicity in human focal cortical dysplasia. METHODS: Immunoblotting and immunoprecipitation were used to quantify these receptor subunits in tissue resected from EEG-verified epileptic and distal nonepileptic frontal cortical areas in each of three patients as determined by chronic subdural electrode recordings. In each patient, adjacent sections were immunostained to verify that the numbers of dysplastic neurons were greater in epileptic than in nonepileptic cortex. RESULTS: In all patients, NMDAR2A/B expressions and their coassemblies with NMDAR1 were increased in epileptic dysplastic cortex compared with the relatively normal appearing nonepileptic cortex. For all three patients, there were no significant differences in NMDAR1 protein expressions between the two EEG groups. CONCLUSIONS: These results suggest that increased NMDAR1-NMDAR2A/B coassembly contributes to hyperexcitability in dysplastic cortical neurons and focal seizure onsets.     

抗-SSA/Ro抗体三种检测方法的比较

目的 比较酶联免疫吸附实验（ELISA）、免疫双扩散（ID）和免疫印迹（IB）3种方法检测抗-SSA/Ro抗体,评价3种方法的符合度,同时提高IB检测相对分子质量52 000条带的认识.方法 采用ELISA法、IB法及ID法检测以下3组对象的抗-SSA/Ro抗体水平：正常献血员血清122份;实验室常规检测血清7736份;诊断明确结缔组织病血清166份.结果 （1）正常献血员122份血清ELISA、ID和IB检测抗-SSA/Ro抗体全部阴性;（2）实验室常规检测的7736份血清,IB检测相对分子质量52 000阳性1085例,其中抗核抗体和其他抗-ENA抗体均阴性的有92份,该组病人经过ID和ELISA两种方法检测全部阴性;（3）结缔组织病组166例,ELISA、ID和IB检测抗-SSA/Ro抗体的阳性率分别是76.5%（127/166）,65.1%（108/166）,49.4%（82/166）;（4）结缔组织病166份血清中,ELISA和ID两种方法检测的符合率88.6%（147/166）,两种方法的比较,χ2=17.1,P＜0.001;ID和IB两种方法检测的符合率为75.9%（126/166）,两种方法的比较,χ2=15.6,P＜0.001;（5）ELISA和ID进行Spearman等级相关分析,相关系数为0.828,P＜0.001.结论 （1）ELISA和ID特异性优于IB.（2）IB特异性较差,但如果同时ANA和或抗-ENA其他条带阳性,特异性会显著提高.（3）ELISA敏感性优于ID,ID敏感性优于IB.（4）ELISA和ID结果在数值上存在显著的正相关关系.     

Cork workers’ occupational asthma: lack of association with allergic sensitisation to fungi of the work environment

Objectives To evaluate allergic sensitisation to Chrysonilia sitophila, Penicillium glabrum, and Trichoderma longibrachiatum in cork workers with asthma.Methods Skin prick tests with a battery of common allergens and with the three fungi were performed on ten cork workers with asthma and eight non-exposed asthmatics. Based on serial peak expiratory flow measurements, five were classified as having occupational asthma (AO) and five as having non-occupational asthma (NOA). In exposed patients, specific antibodies for the three fungi were also studied by immunoblottingResults Two out of ten patients with occupational exposure and four out of eight of the control group showed positive results for skin prick tests for common allergens. Moreover, two out of five patients with OA and three out of eight controls exhibited sensitisation to storage mites. All exposed patients (with OA or NOA) had negative skin prick test results for the fungal extracts. In patients with asthma and occupational exposure, immunoblotting results confirmed the absence of specific IgE. However, specific IgG4 was present in some cases.Conclusions Atopy does not seem to characterise occupational asthma in cork workers. Despite their long exposure to moulds, we could not find evidence of IgE sensitisation to the three most prevalent cork fungi in patients with OA, which points to the search for other causative agents, such as cork chemical compounds or contaminants.     

Improved diagnostic testing for ataxia-telangiectasia by immunoblotting of nuclear lysates for ATM protein expression

The laboratory diagnosis of ataxia-telangiectasia (A-T) currently relies upon measurement of serum alphafetoprotein (AFP) and cellular sensitivity to ionizing radiation. A previous report suggests that immunoblotting of whole cell lysates from lymphoblastoid cell lines (LCLs) might be informative for diagnosis. To further evaluate this possibility, and improve sensitivity, we performed immunoblotting for ATM protein on nuclear lysates of 71 consecutive radiosensitive LCLs that were established from patients with clinical features suggestive of A-T. Fifty-two LCLs (73%) contained no detectable ATM protein, with a representative sample (N=25) testing negative for ATM kinase activity, having at least one ATM mutation, and having elevated AFP levels; these results confirmed the diagnosis. Seventeen LCLs (24%) expressed intermediate or normal levels of ATM protein and exhibited normal ATM kinase activity; follow-up studies failed to detect ATM mutations and AFP levels were normal in all but three. Of the remaining two radiosensitive LCLs, one had 35% of normal protein with normal kinase activity and no ATM mutations. The other LCL had 9% of normal protein, with intermediate levels of kinase activity, a homozygous missense ATM mutation, and elevated AFP. Our data suggest that it is very uncommon to encounter bonafide A-T patients with more than trace amounts of ATM protein. We conclude that immunoblotting for ATM protein is of higher specificity for diagnosing A-T than radiosensitivity testing. In addition, we have documented in vitro radiosensitivity in other patients who share some clinical features with A-T.