采用分子筛层析法分离纯化细粒棘球蚴囊液粗抗原

目的分离纯化细粒棘球蚴囊液抗原,去除其中的非特异性反应蛋白,探索优化囊液抗原制备方法。方法采用超滤法浓缩羊肝细粒棘球蚴囊液制备成粗抗原,并采用免疫印迹实验对囊液抗原进行分析,发现非特异性反应的主要蛋白条带。随后采用Superdex凝胶柱通过分子筛层析法对囊液抗原进行纯化。用44份细粒棘球蚴病患者、17份健康人和10份囊尾蚴患者血清进行酶联免疫实验,评估纯化后囊液抗原的检测能力。结果免疫印迹实验发现,非特异性反应的主要条带的相对分子质量(M_r)约为55 000,纯化后去除了该非特异性反应蛋白。纯化后的囊液抗原ELISA检测灵敏度为93.2%(41/44),特异性为94.1%(16/17),约登指数为0.87,与囊尾蚴病交叉反应性为30%(3/10)。未纯化前的灵敏度为90.9%(40/44),特异性为88.2%(15/17),约登指数0.79,与囊尾蚴病交叉反应性为60%(6/10)。纯化前后ELISA检测的灵敏度存在显著性差异。结论本研究采用分子筛层析法对羊源细粒棘球蚴的囊液抗原进行纯化,去除了引起非特异性反应的主要蛋白(M_r 55 000),为后续进行囊液抗原标准化、提高检测能力提供了依据。     

棘球蚴B抗原序列的不均一性

棘球蚴B抗原是一种耐热的160kDa脂蛋白,为棘球蚴囊液的主要抗原。有约90％细粒棘球蚴病人和约40％泡状棘球蚴病人有其抗体,说明这两种棘球蚴均有B抗原。经免疫荧光检测它主要存在于原头节的表皮细胞,也存在于囊液中,表明它是一种分泌性抗原。 最近已从英国羊获得的细粒棘球蚴中分     

用合成多肽分析细粒棘球蚴AgB抗原亚单位的反应性表位

目的通过合成多肽分析细粒棘球蚴AgB1、AgB2和AgB4等3个亚单位抗原的主要反应性表位区域。方法用ELISA法分析来源于细粒棘球蚴AgB1、AgB2和AgB4亚单位基因序列的5个人工合成多肽KK36、RK30、B4-1、B4-2和B4-3,及上述3个亚单位重组抗原在血清抗体检测中的反应性,共检测细粒棘球蚴病(115例)、多房棘球蚴病(54例)、囊尾蚴病(22例)患者和健康人(18例)血清209份。用受试者工作特征(ROC)曲线分析合成多肽和重组抗原在血清检测中的诊断效率。结果多肽KK36和RK30诊断细粒棘球蚴病的敏感性分别为89.2%和85.0%,特异性为62.5%和59.4%,诊断效率分别为84.8%和80.4%,与AgB1(84.5%)和AgB2(81.2%)抗原诊断效率相近,拟合AgB1和AgB2重组抗原的ROC曲线。来源于AgB4抗原的3个多肽B4-1、B4-2和B4-3检测细粒棘球蚴病患者血清的诊断效率分别为49.4%、57.9%和77.4%。其中,B4-3的反应性最佳,B4-2也有一定的反应性。结论 KK36和RK30完整地包含了AgB1和AgB2抗原的反应性表位区域。B4-2和B4-3多肽...     

检测棘球蚴病患者血清内循环抗原的免疫层析试条的制备和评价

目的 以棘球蚴囊液纯化抗原特异性单克隆抗体为基础建立一种快速、简便诊断棘球蚴病的胶体金免疫层析试条方法,并对其进行评价。方法 纯化棘球蚴囊液抗原,并以此免疫BALB/c小鼠,采用杂交瘤技术制备单克隆抗体,对所制备的单克隆抗体确定其亚类和效价。筛选基于棘球蚴囊液纯化抗原制备的单克隆抗体对,采用柠檬酸三钠还原法制备胶体金颗粒,标记筛选到的单克隆抗体,并将其吸附于交联垫;将另一筛选到的单克隆抗体划线包被于同一硝酸纤维素膜适当位置,制成免疫层析试条。用该试条检测手术确诊的细粒棘球蚴病(87例)、多房棘球蚴病(40例)、囊尾蚴病(25例)、日本血吸虫病(10例)、弓形虫病(5例)、并殖吸虫病(5例)、华支睾吸虫病(5例)患者血清,以及60例健康者血清,以评价其检测的敏感性和特异性。结果 以棘球蚴囊液纯化抗原为免疫源制备单克隆抗体,共筛选了11株能高效分泌效价在1∶25 600～1∶102 400特异抗体的细胞株,抗体亚类为IgG₁或IgG_2a。筛选到的单克隆抗体F₃B₆作为标记抗体,单克隆抗体C_4<...     

羊棘球蚴囊液抗原B诊断早期实验棘球蚴病鼠的研究

采用两种抗原（抗原Ｂ和粗抗原）通过ＥＬＩＳＡ法检测感染细粒棘球蚴和多房棘球蚴２－３月的小鼠,正常鼠以及棘球蚴病人,非棘球蚴病人和健康人血清抗体。结果表明,抗原Ｂ和粗抗原检测２月细粒棘球蚴鼠血清抗体阳性率分别为９５％和１００％。检测３月细粒球蚴病鼠血清抗体均为１００％,检测正常血清的假阳性率分别为０和５％。     

细粒棘球蚴重组抗原B 8-kDa亚单位1对囊型包虫病的血清学诊断价值

目的通过基因工程技术获得细粒棘球蚴抗原B8-kDa亚单位1重组蛋白(rEgAgB8/1),探讨其对囊型包虫病(CE)的血清学诊断价值。方法将构建的rEgAgB8/1原核表达质粒(pET32b-rEgAgB8/1)转化至E.coli BL-21(DE3)中,用IPTG诱导表达,经亲和层析纯化获得高纯度rEgAgB8/1,以rEgAgB8/1为抗原,应用ELISA和Immuno blotting方法对31例手术确诊的囊型包虫病病人血清进行了回顾性检测与分析。结果ELISA和Immuno blotting方法检测CE病人血清阳性率均为90.3%(28/31),3例血清学检测阴性的CE病人均为初次诊断为CE及单纯性肝脏单发感染的病人;血清抗体水平随着病人棘球蚴囊数目增加而有所增加,棘球蚴囊的数目与血清抗体水平的比较用单因素方差分析有显著性差异(F=5.06,P=0.0142),1个囊与2个囊/3个囊组间血清抗体水平有显著差异,2个囊与3个囊组间差异无统计学意义。结论rEgAgB8/1重组蛋白抗原对囊型包虫病有较高的血清学诊断价值,多囊型包虫病人血清抗体水平高于单囊型包虫病人。     

检测包虫病患者血清特异IgG4诊断价值的研究

目的探讨检测患者血清特异IgG4诊断包虫病的效果.方法采用两种抗原(棘球蚴囊液粗抗原及其抗原B)通过ELISA法检测棘球蚴病病人、非棘球蚴病病人和健康对照血清特异IgG4.结果粗抗原和抗原B检测棘球蚴患者血清特异IgG4的阳性率分别为94.4%和89.8%;与部分猪囊尾蚴病患者血清出现交叉反应外,与肺吸虫病、旋毛虫病、血吸虫病、肝囊肿等患者的血清以及健康对照血清均未出现交叉反应.结论检测棘球蚴患者血清特异IgG4敏感性高,特异性强,具有较好的诊断价值.     

云南腾冲市棘球蚴病线索调查

目的了解腾冲市棘球蚴病的感染现状,为制订棘球蚴病防治策略提供依据。方法以2011～2017年报告病例为线索,选取报告病例所在自然村及毗邻的2～4个村,对1岁以上常住居民进行腹部B超检查,开展人群棘球蚴患病情况调查。对报告病例所在小学的全体学生进行腹部B超检查,同时采集静脉血,ELISA试剂盒检测血清抗棘球蚴Ig G抗体水平。采用内脏剖检法调查牲畜和啮齿动物棘球蚴感染情况。采集报告病例所在自然村的所有家犬犬粪,检测犬粪棘球蚴抗原。结果进行B超检查3 153人,未发现棘球蚴病患者;检测血清376份,血清抗棘球蚴Ig G抗体阳性率为0.27%(1/376);中间宿主检查牛11头、羊113只、猪2 217头,鼠48只,未发现棘球蚴感染;检测终宿主犬粪251份,犬粪棘球蚴抗原阳性率为0.4%(1/251)。结论腾冲市部分乡镇存在棘球蚴病流行,应加强传染源犬的监测和管理。     

3种抗棘球蚴抗体检测试剂盒用于棘球蚴病血清学诊断效果评价

目的　评价我国3种商品化抗棘球蚴抗体检测试剂盒用于棘球蚴病血清学诊断的效能。方法　收集142份细粒棘球蚴病、89份多房棘球蚴病确诊患者及39份健康对照者血清标本,分别采用试剂盒A[酶联免疫吸附试验（ELISA法）]、B（ELISA法）、C（胶体金法）检测,比较细粒棘球蚴病和多房棘球蚴病患者血常规和生化指标差异,分析采用试剂盒A和B检测的棘球蚴病患者血清特异性抗体吸光度值（A值）与血常规及生化指标的相关性,评价3种试剂盒检测棘球蚴病的效果。结果　细粒棘球蚴病和多房棘球蚴病患者白细胞计数（WBC）、中性粒细胞计数（NEU）、单核细胞计数（MONO）、嗜碱性粒细胞计数（BASO）、丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、直接胆红素（DBIL）、间接胆红素（IBIL）中位数差异无统计学意义（P均 > 0.05）,细粒棘球蚴病患者淋巴细胞计数（LYM）和白蛋白（ALB）中位数显著高于多房棘球蚴病患者（P均< 0.05）,而多房棘球蚴病患者嗜酸性粒细胞计数（EOS）中位数显著高于细粒棘球蚴病患者（P < 0.01）。采用试剂盒A检测的棘球蚴病患者血清特异性抗体A值与患者WBC（rs = 0.153,P < 0.05）、EOS（rs = 0.174,P < 0.05）呈线性正相关,与TBIL（rs = -0.134,P < 0.05）、IBIL（rs = -0.146,P < 0.05）呈线性负相关。采用试剂盒B检测的棘球蚴病患者血清特异性抗体A值与WBC（rs = 0.257,P < 0.01）、NEU（rs = 0.203,P < 0.01）、MONO（rs = 0.159,P < 0.05）、EOS（rs = 0.330,P < 0.01）、ALT（rs = 0.171,P < 0.01）、AST（rs = 0.160,P < 0.05）呈线性正相关,与ALB呈线性负相关（rs = -0.168,P < 0.05）。试剂盒A、B和C诊断棘球蚴病患者的总符合率分别为86.30%、69.63%和91.48%,敏感度分别为84.42%、64.94%和92.21%,特异度分别为97.44%、97.44%和87.18%,约登指数分别为0.82、0.62和0.79,Kappa值分别为0.600、0.337和0.750。试剂盒A、B和C诊断细粒棘球蚴病患者的总符合率分别为84.54%、64.64%和71.82%,敏感度分别为80.99%、55.63%和68.31%,特异度分别为97.44%、97.44%和87.18%,约登指数分别为0.78、0.53和0.56;试剂盒A、B和C诊断多房棘球蚴病患者的总符合率分别为92.19%、85.16%和85.16%,敏感度分别为89.89%、79.78%和84.27%,特异度分别为97.44%、97.44%和87.18%,约登指数分别为0.87、0.77和0.72。试剂盒C在诊断细粒和多房棘球蚴病时存在交叉反应。试剂盒A和B用于诊断棘球蚴病的受试者工作特征曲线（receiver operating characteristic curve,ROC）曲线下面积分别为0.970和0.948,差异无统计学意义（Z = 1.618,P > 0.05）,试剂盒A和B诊断棘球蚴病结果具有较好一致性（Kappa = 0.585,P < 0.01）。结论　3种抗棘球蚴抗体检测试剂盒对多房棘球蚴病血清学诊断效果均优于细粒棘球蚴病。试剂盒A诊断棘球蚴病具有较高敏感性和特异性,检测性能相对稳定、干扰因素相对较少,适合用于棘球蚴病术前初步诊断和术后患者血清抗体随访监测;试剂盒C诊断棘球蚴病具有较高敏感性和特异性,操作方便、报告率高,适用于棘球蚴病初步筛查。     

细粒棘球蚴感染对小鼠炎症性肠病的保护机制研究

目的 观察细粒棘球蚴感染对葡聚糖硫酸钠(DSS)诱导的小鼠炎症性肠病(IBD)症状的保护作用及可能机制。方法 建立继发性细粒棘球蚴感染小鼠模型,再给予DSS诱导IBD症状,解剖后ELISA法检测小鼠血清中囊液抗原水平,并观察腹腔内的包囊;根据每日体重监测、解剖后结肠长度测量及结肠组织病理学评分指标评估细粒棘球蚴感染对炎症性肠病的保护作用;采用流式细胞术CBA法检测模型鼠血清中Thl/Th2/Thl7水平,评价细胞因子对炎症性肠病的保护作用。结果 小鼠感染细粒棘球蚴后血清HCF抗体阳性率与成囊率均为100%。细粒棘球蚴感染减轻了DSS引起的小鼠体重下降程度;结肠变短程度减小,HE染色检查小鼠结肠组织炎性症状显著改善。CBA检测显示细粒棘球蚴感染小鼠血清IL-4、IL-10、IL-17A、IL-6、IFNγ含量均较正常小鼠显著增加,表现为Th2为主的免疫反应;细粒棘球蚴感染合并IBD模型组小鼠Th2/Th1型细胞因子比率同IBD组相比显著升高。结论 细粒棘球蚴感染IBD模型小鼠血清IL-4、IL-10水平升高,Th2高水平状态削弱了DSS引发的Th1反应,表明细粒棘球蚴感染可改善IBD小鼠...     

Split Right Coronary Artery Its Definition and Its Territory

We report here, for perhaps the 1st time in the English-language literature, the extent of the territory fed by the anterior bifurcation of the (anomalous) split right coronary artery (RCA). A 64-year-old man presented with an occlusion of the anterior bifurcation of a split RCA—which resulted in an infarct that involved both the inferoseptal left ventricular wall and the anterior right ventricular free wall. Split RCA is the same anomaly as the improperly named “double right coronary artery.” In reality, there are not 2 RCAs, but only split portions of the posterior descending branch of the RCA, with 2 separate proximal courses.Key words: Coronary vessel anomalies/diagnosis, myocardial infarction/diagnosis/physiopathologyIn December 2007, a 64-year-old hypertensive man, a chronic smoker with dyslipidemia, presented at the hospital 6 hours after the onset of chest pain. In the emergency room, an electrocardiogram (ECG) showed a pre-existing right bundle branch block with new Q waves in the inferior leads and ST-segment elevation in the anterior precordial leads (Fig. 1). Cardiac enzymes were elevated (creatine kinase, 702 U/L; creatine kinase MB, 91.1%; cardiac troponin T, 0.344 ng/mL). An echocardiogram on admission showed basal inferior and mid-inferior akinesia with anterior right ventricular wall hypokinesia. The anterior left ventricular wall contracted normally.Open in a separate windowFig. 1 Electrocardiogram on admission.Due to the presumptive diagnosis of acute anterior myocardial infarction, the patient was immediately studied by means of coronary angiography, which showed a normal left coronary artery (Fig. 2), a split right coronary artery (RCA) with proximal occlusion of the anterior bifurcation, and no distal filling through collateral vessels (Fig. 3). The posterior subdivision of the RCA led to a small upper posterior descending branch and a large posterolateral branch (Fig. 3). The occluded anterior subdivision was entered and dilated; a 2.75 × 14-mm Endeavor® stent (Medtronic, Inc.; Minneapolis, Minn) was deployed, which resulted in recovery of TIMI-3 flow while antegrade filling of the posterior descending branch showed on angiography (Fig. 4). Follow-up ECGs showed gradual resolution of the ST-T changes (Fig. 5).Open in a separate windowFig. 5 Two days after angioplasty, an electrocardiogram shows that ST-T changes are beginning to resolve.Open in a separate windowFig. 4 Immediately after angioplasty, antegrade filling of the more distal posterior descending branch appears upon angiography.Open in a separate windowFig. 3 Angiogram on admission shows total occlusion (arrow) of the split anterior bifurcation of the right coronary artery.Open in a separate windowFig. 2 Angiogram on admission shows near-normal left coronary artery.     

Urotensin II: Its Function in Health and Its Role in Disease

Urotensin II (U-II) is the most potent vasoconstrictor known, even more potent than endothelin-1. It was first isolated from the fish spinal cord and has been recognized as a hormone in the neurosecretory system of teleost fish for over 30 years. After the identification of U-II in humans and the orphan human G-protein-coupled receptor 14 as the urotensin II receptor, UT, many studies have shown that U-II may play an important role in cardiovascular regulation. Human urotensin II (hU-II) is an 11 amino acid cyclic peptide, generated by proteolytic cleavage from a precursor prohormone. It is expressed in the central nervous system as well as other tissues, such as kidney, spleen, small intestine, thymus, prostate, pituitary, and adrenal gland and circulates in human plasma. The plasma U-II level is elevated in renal failure, congestive heart failure, diabetes mellitus, systemic hypertension and portal hypertension caused by liver cirrhosis. The effect of U-II on the vascular system is variable, depending on species, vascular bed and calibre of the vessel. The net effect on vascular tone is a balance between endothelium-independent vasoconstriction and endothelium-dependent vasodilatation. U-II is also a neuropeptide and may play a role in tumour development. The development of UT receptor antagonists may provide a useful research tool as well as a novel treatment for cardiorenal diseases.     

Alcoholics Anonymous and Its Finances

The financial structure of Alcoholics Anonymous is examined in this study through an analysis of its activities in some European countries and the U.S. A focus on the interrelational between the material and the spiritual, in light of the philosophy and traditions of Alcoholics Anonymous, guides the discussion.     

卒中相关性肺炎及其发病机制

卒中相关性肺炎（SAP）是卒中患者病情恶化和死亡的主要原因之一，其发生和发展与卒中患者的基础状态、卒中部位、严重程度、全身和呼吸系统局部免疫功能下降以及不恰当的治疗等因素有关。细菌定植、误吸和机体抵抗力减弱是SAP发病机制中的重要环节。     

Human Gut Microbiota and Its Metabolites Impact Immune Responses in COVID-19 and Its Complications

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