M2受体自身抗体在重度子痫前期患者中的表达

目的：检测M2受体自身抗体在重度子痫前期患者中的表达,探讨其临床意义。方法：选择2013年1月至2015年1月在首都医科大学附属朝阳医院妇产科收治的78例重度子痫前期孕妇为研究对象,并取同期收治的78例正常单胎足月孕妇和60例未妊娠妇女作为对照。采用ELISA方法检测3组妇女血浆中的M2受体自身抗体水平,并分析其在重度子痫前期中的临床意义。结果：重度子痫患者M2受体自身抗体阳性率32.1%(25/78),正常妊娠组为10.3%(8/78),两组阳性率比较差异有统计学意义(P<0.01);未妊娠组阳性率8.3%(5/60),与正常妊娠组比较差异无统计学意义(P>0.05);此外,重度子痫前期患者血清中肌酸激酶浓度为(101.49±142.75) U/L,而正常妊娠组肌酸激酶含量为(57.94.±31.64) U/L,两组比较差异有统计学意义(P<0.05)。结论：M2受体自身抗体在重度子痫前期患者中的表达明显增高,可能与其发病有关。     

人肌红蛋白原核表达载体的构建、表达及蛋白纯化

目的：构建人肌红蛋白（Mb）原核表达载体并诱导其表达、纯化及鉴定目的蛋白pQE。方法：根据已报道的人Mb基因序列,采用RT-PCR技术从人骨骼肌组织中获得肌红蛋白cDNA序列。将所得的PCR产物插入克隆载体pGEM-Teasy中,得重组质粒,经酶切、PCR及测序鉴定正确后,将酶切目的片段插入原核表达载体pQpQEE30中并转化大肠杆菌(E.coli)M15,通过IPTG诱 导表达出带有6个组氨酸的融合蛋白。经镍固定金属亲和层析纯化。结果：序列分析表明,人Mb基因成熟肽编码区含有465 bp,编码153个氨基酸;与GenBank(NM203378) 中已报道的Mb核苷酸序列有98.50 %同源性。经IPTG诱导表达,SDS-PAGE 电泳和免疫印迹分析显示,表达的融合蛋白占菌体蛋白总量的31%,相对分子质量约为16 700,并与商品化的人Mb单抗呈特异性反应。经Ni²⁺-NTA agarose 纯化获得SDS-PAGE 电泳下单一条带。结论：在大肠杆菌中获得了人Mb的高效表达。     

Circulating autoantibodies against beta-adrenergic receptors in patients with heart diseases]

Patients with different heart diseases, dilated cardiomyopathy, valvular heart disease, hypertension, ischemic heart disease or myocarditis showed manifestations of autoimmunity and down-regulation of beta-adrenergic receptors. Autoantibodies against beta-adrenergic receptors in these patients were detected with radioligand binding inhibition assay. The results suggested that the down-regulation of cardiac beta-adrenergic receptors in these patients may be mediated by autoimmunity. Autoantibodies against beta-adrenergic receptor were not related to any specific heart diseases, but to the severity of heart failure irrespective of its etiology. The significance of these autoantibodies in heart failure was discussed.     

抗β2-AR肾上腺素受体自身抗体功能的初步研究

目的研究抗β2-肾上腺素受体(β2-adrenoceptor,β2-AR)自身抗体对乳鼠心肌细胞跳动频率的影响。方法①手术结扎大鼠冠状动脉建立心梗模型;②在大鼠心力衰竭(心衰)的不同时期采用链霉亲和素-酶联免疫吸附法(SA-ELISA)检测大鼠血清中抗β2-AR自身抗体的滴度,同时检测大鼠的心功能;③将β2-AR激动剂硫酸特布他林、抗β2-AR自身抗体的阳性血清及抗β2-AR自身抗体 β2-AR组的血清加至培养的乳鼠心肌细胞中,观察该自身抗体对乳鼠心肌细胞跳动频率的影响。结果①将β2-AR激动剂硫酸特布他林(4.5μmol/L)加入培养的乳鼠心肌细胞中,加药前心肌细胞跳动频率(次/15 s)为20.0±6.20,加入后第5,15和30 min分别为16.92±4.89,13.17±3.21和11.85±2.84。②将大鼠抗β2-AR自身抗体的阳性血清加至培养的乳鼠心肌细胞中,加样前心肌细胞跳动频率(次/15 s)为26.43±3.63,加入后第5,15和30 min分别为9.42±3.03,10.00±1.76和9.25±2.56,心肌细胞跳动频率减慢,且在观察时间内不随时间递减,表现出类激动剂样生物效应。③大鼠抗β2-AR自身抗体 β2-AR组的血清加至培养的乳鼠心肌细胞中,加样前心肌细胞跳动频率(次/15 s)为21±3.4,加入后第5,15和30 min分别为18.9±2.72,17.6±2.48和16.1±2.9,心肌细胞的跳动频率无明显减慢。结论抗β2-AR自身抗体作用于培养的乳鼠心肌细胞,可使其跳动频率减慢,且在观察时间内不随时间递减,表现出类激动剂样生物学效应,加入相应抗原后基本可中和其效应。     

抗心肌ADP/ATP载体对小儿心脏病的诊断价值

目的 :探讨抗心肌ADP/ATP载体 (ANT)抗体对小儿心脏病的诊断价值。方法 :运用免疫印转法检测了 3 0例病毒性心肌炎 (VMC)、14例扩张型心肌型 (DCM) ,10例风湿性心脏病 (RHD)、2 2例先天性心脏病 (CHD)患儿血浆抗ANT水平。结果 :抗ANT抗体在VMC和DCM患儿中的阳性率显著高于RHD、CHD和NC三组研究对象 (P均 <0 .0 1)。抗ANT抗体诊断VMC和DCM的敏感性为 72 73 % ,特异性为 98 2 5 %。结论 :抗ANT抗体在小儿心脏病的鉴别诊断中有重要价值 ,可作为小儿VMC和DCM的特异性诊断指标     

黑质自身抗体在MPTP致小鼠帕金森病发病中的作用

目的探讨MPTP致小鼠帕金森病PD发病中自身抗体是否存在及可能的作用。方法使用MPTP亚急性模型建立PD模型。采集建模后小鼠血清与自身黑质蛋白质相互作用,通过免疫组织化学、酶链免疫吸附测定（ELISA）、蛋白免疫印迹法（Western blot）等方法探讨该PD模型有无自身抗体的存在,并初步推断自身抗体是否参与PD发病过程。结果 MPTP致PD模型中,血清和自身黑质的结合性增加。结论 MPTP致PD模型中,血清中可能存在针对黑质的自身抗体,参与了PD的发病过程。     

A novel autoantibody in patients with primary hypertension： antibody against L-type Ca^2＋ channel

Background Recently, it has been proposed that the autoantibodies against various cardiovascular receptors play a role in the pathogenesis of primary hypertension. In this study, we aimed to identify whether or not there are autoantibodies against cardiovascular L-type Ca^2＋ channels in patients with primary hypertension. Methods A peptide corresponding to the sequence 2-16 of the alc-subunit of L-type Ca^2＋ channel was used as an antigen to screen the autoantibodies from 90 patients with primary hypertension and 45 healthy controls by an enzyme-linked immunosorbent assay （ELISA）. The clinical data of 90 hypertensive patients were compared between patients with and without these autoantibodies. Results Serum from 3 （6.7%） of the 45 healthy controls, 33 （36.7%） of 90 hypertensives showed positive responses in ELISA （P 〈0.01）. The prevalence of such autoantibodies in two subgroups of hypertensives with coronary heart disease （9/21, 57.14%, P 〈0.05） and left ventricular diastolic dysfunction （28/63, 44.4%, P 〈0.05） was higher than in those without the corresponding complications. And the patients with such autoantibodies had lower E/A than patients without such autoantibodies （0.803±0.191 vs 1.004±0.322, P=0.002）. Conclusion There are autoantibodies against vascular L-tyPe Ca^2＋ channels in patients with primary hvnertension.     

Antiviral Activity of Dual-acting Hydrocarbon-stapled Peptides against HIV-1 Predominantly Circulating in China

Objective New rationally designed i,i+7-hydrocarbon-stapled peptides that target both HIV-1 assembly and entry have been shown to have antiviral activity against HIV-1 subtypes circulating in Europe and North America. Here, we aimed to evaluate the antiviral activity of these peptides against HIV-1 subtypes predominantly circulating in China. Methods The antiviral activity of three i,i+7-hydrocarbon-stapled peptides, NYAD-36, NYAD-67, and NYAD-66, against primary HIV-1 CRF07 BC and CRF01 AE isolates was evaluated in peripheral blood mononuclear cells (PBMCs). The activity against the CRF07 BC and CRF01 AE Env-pseudotyped viruses was analyzed in TZM-bl cells. Results We found that all the stapled peptides were effective in inhibiting infection by all the primary HIV-1 isolates tested, with 50% inhibitory concentration toward viral replication (IC50) in the low micromolar range. NYAD-36 and NYAD-67 showed better antiviral activity than NYAD-66 did. We further evaluated the sensitivity of CRF01 AE and CRF07 BC Env-pseudotyped viruses to these stapled peptides in a single-cycle virus infectivity assay. As observed with the primary isolates, the IC50s were in the low micromolar range, and NYAD-66 was less effective than NYAD-36 and NYAD-67. Conclusion Hydrocarbon-stapled peptides appear to have broad antiviral activity against the predominant HIV-1 viruses in China. This finding may provide the impetus to the rational design of peptides for future antiviral therapy.     

流式免疫微球芯片技术检测血小板膜糖蛋白特异性自身抗体的方法及其临床应用

目的 建立流式免疫微球芯片技术(FCIBA)检测血小板膜糖蛋白(GP)特异性自身抗体的方法及其临床应用.方法 使用流式细胞仪检测包被不同GP的单克隆抗体,抗GPⅨ(SZ1)、抗GPⅠb(SZ2)、抗GP Ⅲa(SZ21)、抗GPⅡb(SZ22)、抗P-选择素(SZ51)的不同荧光强度的微球,检测结果与单克隆抗体俘获血小板抗原技术(MAI-PA)进行比较.选取特发性血小板减少性紫癜(ITP)组、非ITP (NITP)组和健康人对照组样本用FCIBA分析SZ1、SZ2、SZ21、SZ22、SZ51共5种特异性血小板自身抗体.结果 批内批间差异小;rTP组与NITP组和健康人对照组比较,差异均有高度统计学意义(均P ＜0.01);对ITP诊断的总特异度为81.4％(70/86),MAIPA总特异度为80.2％(69/86),差异无统计学意义(P＞0.05).敏感度为62.00％ (31/50),明显高于改良间接MAIPA的31.00％(19/50,P＜0.05).结论 FCIBA可以一次同时进行5种自身抗体的联合检测,为ITP的临床诊断提供了一种快速测定的新方法.     

粘附分子在多发性肌炎中的表达

目的：探讨粘附分子参与多发性肌炎(PM)的病理机制。方法：采用直接双色荧光免疫标记法和RT—PCR法检测PM患者外周血淋巴细胞表面和肌组织中粘附分予对应配体的异常表达。结果：PM患者外周血淋巴细胞表面粘附分子淋巴细胞功能抗原1(LFA—1)、极迟抗原4(VLA-4)和肌肉组织细胞间粘附分子1(ICAM—1)mRNA的表达均明显高于正常对照组。结论：LFA—1、VLA-4和ICAM—1在多发性肌炎患者中呈高表达，提示粘附分子在PM的炎性细胞浸润破坏病变中可能起介导作用。     

应用细胞内PCR技术构建及筛选白癜风噬菌体抗体库

目的: 克隆白癜风患者淋巴细胞中原始配对的自身抗体基因,构建噬菌体抗体库并筛选针对黑素细胞膜抗原的特异性抗体. 方法: 细胞内PCR法扩增人免疫球蛋白轻链和重链可变区基因,Cre重组酶介导下完成单个细胞内的轻重链连接,获得反映体内原始配对信息的单链抗体可变区基因,构建噬菌体抗体库,筛选针对黑素细胞膜抗原的自身抗体. 结果: 构建完成库容量约为3×l06的噬菌体抗体库,筛选得到1株针对黑素细胞膜抗原的自身抗体. 结论: 利用细胞内PCR技术和噬菌体抗体库技术成功构建免疫文库,为进一步研究自身抗体在白癜风发生发展过程中的作用奠定了基础.     

Autoantibodies against human insulin

Sera from 680 non-diabetic subjects with suspected autoimmune disease were screened for 13 different antibodies. Of the 582 sera found to contain these antibodies, nine bound insulin in an IgG specific enzyme linked immunosorbent assay (micro ELISA). Four of the sera bound human, porcine, and bovine insulins and five bound exclusively human insulin. "Cold" human, porcine, and bovine insulins each displaced, in a dose dependent manner, the four sera which bound all three insulins, but only human insulin displaced the remaining five, porcine and bovine insulins having little or no effect in concentrations up to 1000 U/1. These observations point to the existence of autoantibodies specifically against human insulin in some subjects with established autoimmunity.     

皮肌炎/多肌炎患者血清对血管内皮细胞的影响

目的:探讨皮肌炎/多肌炎(dermatomyositis/polymyositis,DM/PM)患者血清对血管内皮细胞活性的影响.方法:取16例未经治疗的DM/PM患者血清为DM/PM组,其中8例治疗前取血清作为治疗前组,经治疗病情好转后再次取血清作为治疗后组,13名健康体检者血清为正常组.体外培养脐静脉内皮细胞(hu...