Der f 1血清特异性IgE ELISA检测方法的建立

目的利用粉尘螨变应原第1组分重组蛋白rDer f 1建立特异性IgE ELISA检测方法,并对哮喘患者血清进行检测。方法以rDer f 1为包被抗原,分别建立间接ELISA法和亲和素-生物素复合ELISA(ABC-ELISA)法,检测Der f 1特异性IgE阳性标准血清和39例哮喘患者血清中的Der f 1特异性IgE,并分析检测结果。结果间接ELISA法和ABC-ELISA法检测Der f 1特异性IgE的最适抗原包被浓度和血清稀释倍数均为15μg/ml和1︰2,间接ELISA法辣根过氧化物酶(HRP)标记抗人IgE抗体适宜稀释倍数为1︰1 000,ABC-ELISA法生物素标记抗人IgE抗体和HRP标记亲和素的稀释倍数分别为1︰1 000和1︰2 000。用间接ELISA和ABC-ELISA检测IgE的灵敏度分别为0.68U/ml和0.73U/ml,检测患者血清中Der f 1特异性IgE的阳性率分别为92.3%和97.4%,差异无统计学意义(P>0.05)。结论基于重组变应原rDer f 1的ELISA法检测粉尘螨特异性IgE具有较高的敏感度,可用于螨过敏性疾病的实验诊断。     

ELISA测定粉尘螨特异性IgE、IgG和总IgE的研究

用ELISA间接法测定粉尘螨(Dermatophagoidesfarinae)特异性IgE和特异性IgG,双抗体法测定总IgE。实验中采取棋盘滴定,选择了包被的抗原或抗体、血清和酶结合物的最适浓度,并对抗原包被     

ABC-ELISA法诊断肠螨病的研究

目的　探讨生物素 -亲和素酶联免疫吸附试验 (ABC -ELISA)在肠螨病诊断中的应用价值。方法　采集 4 8例肠螨病患者血清 ,用ABC -ELISA法检测血清中螨特异性抗体IgG ,并与葡萄球菌A -蛋白酶联免疫吸附试验 (SPA -ELISA)进行比较。结果　用ABC -ELISA法和SPA -ELISA法检测 4 8例肠螨病患者血清中的螨特异性抗体IgG阳性率分别为89 5 8% (43/ 4 8)和 5 6 2 5 % (2 7/ 4 8) ,两者比较 ,差异具显著性 (χ2 =13 5 0 ,P <0 0 1)。结论　ABC -ELISA法是实验室诊断肠螨病的一种有效方法     

应用重组抗原(rSAG1)检测IgM抗体诊断弓形虫病的研究

目的以刚地弓形虫RH株重组主要表面抗原1作为检测抗原,建立检测弓形虫IgM抗体的rSAG1-ELISA.方法用Ni2 螯合柱纯化重组抗原rSAGI;以不同浓度的rSAG1的包被聚苯乙烯酶标条,阳性和阴性混合血清分别作1:100和1:200稀释,以辣根过氧化物酶标记的羊抗人IgM为二抗,采用正交试验确定rSAG1-ELISA的最佳检测条件;按其最佳检测条件分别对弓形虫IgM阳性和阴性混合血清重复测20次进行精确度的检测;采用抗体抑制试验检测其特异性;同时对用进口试剂盒筛得的35份弓形虫IgM阳性血清和57份弓形虫IgM阴性血清进行检测.结果 制备的rSAG1重组蛋白纯度在90%以上;rSAG1-ELISA的最佳检测条件为:rSAG1的包被浓度为2.5μg/m1,血清稀释度1:100,酶标记的羊抗人IgM 1:4 000稀释;用rSAG1-ELISA对混合弓形虫IgM阳性和阴性血清的重复检测表明:IgM阳性混合血清检测值的变异系数(CV值)为13.8%,IgM阴性混合血清的CV值为7.7%;灵敏度检测表明血清稀释度在1:50～1:200均可检出阳性;特异性试验的抑制率为62.0%;rSAG1-ELISA与进口试剂盒的总符合率为88.0%,其中阳性和阴性符合率分别为82.9%(29/35)和91.2%(52/57).结论 rSAG1-ELISA具有较好的灵敏度和特异性,与进口试剂盒相比有较高的符合率,表明该法对弓形虫病早期诊断具有较高的价值,可进一步推广应用.     

ELISA法检测血清中抗CNN2抗体的实验条件优化性探讨

目的 探讨血清抗钙调节蛋白Calponin 2(CNN2)抗体的最适酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)实验条件。方法 将BL21(DE3) p Lys S-pColdⅢ-C基因工程菌重组表达而分离纯化的CNN2包被至酶标板。采集肝细胞癌患者血清和正常人血清,探讨蛋白包被浓度、血清稀释液的选择、封闭方式及条件的选择、HRP标记亲和素稀释度的最适条件。结果 CNN2蛋白浓度为10μg/ml时为最适蛋白包被浓度。0. 5%胎牛血清蛋白(BSA)为最适封闭方式。使用PBST来稀释血清,成本低,效果好。亲和素的最适稀释比为1∶9 000。最适实验条件的重复性良好。ROC曲线分析结果显示,最适实验条件下,该检测方法 ROC曲线下面积(AUC)为0. 978(95%CI:0. 947～1. 000,P=0. 000);截断值为0. 484,其对应的灵敏度为96. 0%,特异度为93. 0%。结论 构建了检测血清抗CNN2抗体的最适ELISA实验条件。     

粉尘螨浸液免疫治疗螨性哮喘患者的免疫功能观察

目的　探讨粉尘螨变应原浸液免疫治疗对螨性哮喘患者免疫功能的影响。　方法　用粉尘螨浸液和平喘药分别对实验组和对照组螨性哮喘患者进行免疫治疗和对症治疗 ,用ELISA法检测患者治疗前、后血清总IgE、螨特异性IgE、螨特异性IgG、IL 2和IL 4水平 ,用生物素 链霉亲和素法检测患者治疗前、后外周血CD3 + 、CD4+ 、CD8+ 和CD4+ /CD8+ 。　结果　患者血清总IgE、螨特异性IgE、螨特异性IgG治疗前分别为 (2 92 .3 5± 112 .46)IU/ml、(2 5 1.68±12 5 .15 )IU /ml和 (5 87.64± 3 5 4.68)IU /ml ,治疗后分别为 (2 65 .74± 12 4.67)IU /ml、(2 97.5 6± 172 .2 7)IU /ml和(82 4.5 1± 42 8.2 6)IU /ml,两者相比 ,螨特异性IgG显著上升 (P <0 .0 1)。外周血CD3 + 、CD4+ 、CD8+ 、CD4+ /CD8+ 治疗前分别为 (5 5 .87± 7.2 3 ) %、(3 8.43± 6.43 ) %、(3 0 .14± 5 .2 4) %和 1.2 6± 0 .5 6,治疗后分别为 (65 .83± 6.5 5 ) %、(4 2 .72± 6.2 6) %、(2 8.5 7± 4.67) %和 1.5 8± 0 .62 ,CD4+ /CD8+ 比值上升 ,差异显著 (P <0 .0 5～P <0 .0 1) ;治疗前、后IL 2和IL 4的水平分别为 (2 .16± 0 .3 8) pg/ml、(3 .49± 0 .5 7) pg/ml和 (1.64± 0 .82 )pg/ml、(1.0 3± 0 .78) pg/ml ,差异均具显著性 (P <0 .0     

合成肽作抗原酶免疫技术检测非甲-戊型肝炎病人血清中抗HGV

本文应用人工合成肽作抗原建立了检测庚型肝炎病毒(HGV)抗体的酶免疫技术(EIA),最适抗原包被浓度为4ug/ml。在特异性验证的基础上检测34例非甲-戊5型肝炎病人血清和32例丙型肝炎感染病人血清。结果表明,在     

应用ABC-ELISA检测包虫病人血清特异性IgE

本文报告了应用一种简单而敏感的酶免疫分析法——ABC(Avidin-Biotin-HRP Complex)-ELISA检测包虫病人和猪囊虫病人血清总IgE含量,以常规ELISA和ABC-ELISA检测病人血清抗包虫特异性IgE。 多种蠕虫病患者的血清IgE抗体升高、嗜酸粒细胞增多,皮肤呈速发型超敏反应。人体血清中总IgE含量甚少,特异性IgE含量更少,故检测方法的     

西尼罗病毒抗体酶联免疫吸附试验检测方法的评价

目的系统地评价血清西尼罗病毒(West Nile virus,WNV)抗体的酶联免疫吸附试验(ELISA)检测方法,为在人群和宿主动物中进行WNV感染血清流行病学调查提供技术支持。方法利用构建的包膜蛋白重组质粒(pQE-30)表达纯化西尼罗病毒包膜蛋白,以此为抗原进行ELISA检测。在对抗原包被量、酶标抗体浓度、血清稀释度进行优化的基础上,对本方法的灵敏度和特异度进行评价。结果确定最适抗原包被量为0.034μg,酶标抗体工作浓度和血清稀释度分别为1∶4 800和1∶80;批内变异和批间变异分别为6.7%和22.6%;对小鼠WNV抗体阳性血清53份、JEV抗体阳性血清48份和阴性对照血清94份进行检测,灵敏度为86.8%,特异度分别为93.8%和92.6%。结论本研究建立的ELISA方法灵敏、检测抗体特异,结果可重复,是一种有价值的血清学调查方法。     

粉尘螨特异性IgE对皮肤挑刺试验和鼻腔激发试验的标准化

在吸入型哮喘患者中,进行了粉尘螨Dermato-phagoides farinae浸液的皮肤挑刺试验、鼻腔激发试验和血清特异性IgE的测定。以无哮喘、无过敏性鼻炎的健康者为对照。结果表明皮肤试验的反应率与所取的风团直径有关,鼻腔试验的累积阳性率与所使     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.