IFA诊断黑热病所用实验材料的检测效果评价

IFA虽已常规用于黑热病的诊断和流行病学调查,但对IFA试验中实验材料的选择可能给检测带来的影响及影响的程度等却无较系统的探讨。为此,我们比较了这些因素对检测结果的影响。 用3株杜氏利什曼原虫制备前鞭毛体抗原:(1)保存人株抗原:自四川南坪一例病人骨髓分离。前鞭毛体经NNN培养基培养10天,收集培养物制成抗原片后放-20℃中保存31个月。(2)新鲜人株抗原:自汶川     

恙虫病东方体Sta56和Sta58抗原的研究进展

引起恙虫病的病原体——恙虫病东方体(0rientia tsutsugamushi，Ot．)是一种生长缓慢、严格专性细胞内寄生的革兰氏阴性微生物，分类学上属于立克次体科恙虫病东方体属。纯化的Ot．在SDS-PAGE上出现约30条多肽带，其中56kD抗原和58kD抗原分别为Ot．外膜和细胞内含量最丰富的蛋白。本文就Ot．这两种抗原研究情况作简要介     

培养利什曼原虫的一种改良的液体血琼脂培养基

Difco血琼脂B45培养基,对巴西利什曼种团和利什曼原虫的培养是有用的,可为血清学研究提供大量的巴西利什曼抗原。本文作者以血琼脂为基础,发展了一种液体的B45培养基。将血琼脂基置于-20℃冰冻12小时,而后加无菌蒸馏水稀释成17%浓度, 浸于自来水或置37℃水溶中融化,以无菌操作按1～3ml量分装于无菌的试管内,在-20℃内可保存一年。作者指出,巴西利什曼原虫在这种液体培养基内生长良好,与在常规血琼脂斜面冷凝水内的生长一样,同时在玻璃维亚用此种     

悬浮培养保存恶性疟原虫的一种简单自动装置

鉴于恶性疟原虫能在悬浮培养中生长,并可获得较高的原虫密度。本文作者设计了一种简单的自动培养装置。保存于6℃中的培养基贮存器,以不锈钢管通过螺旋盖与37℃标准轨道孵箱中的培养瓶连接。培养瓶为250ml三角烧瓶,内盛50ml培养液,另装不锈钢管,供气体和更换培养基的进出口与废料瓶及标本收集瓶连接,另一为取样管伸至瓶底,能吸出大量培养基,而不吸出红细胞。其外端装一只疫苗瓶或取样瓶(用于取样或加入新鲜红细胞)。培养基的进出由两个蠕动泵操作。轨道孵育箱     

大肠杆菌O157∶H7 CVa9株O抗原变异的研究

目的　研究本实验室保藏的大肠杆菌O15 7:H7(E coliO15 7:H7)日本分离株CVa9O抗原的变异。方法　血清凝集试验检测CVa9菌株O和H抗原 ,区分O抗原变异株与非变异株。聚合酶链反应 (PCR)法分别检测O15 7和H7特异性基因。生化试验检测生化特性。观察菌株在麦康凯、山梨醇麦康凯、棉子糖麦康凯及ChromagarO15 7显色培养基上菌落形态 ,菌落计数法计算变异率。结果　抗 -O15 7抗血清与CVa9菌株进行玻片凝集试验出现强凝集 (CVa9- 8)和不凝集 (CVa9- 1、CVa9- 15 )两种菌落 ,试管凝集试验结果相同 ,证实不凝集菌落O抗原发生变异。抗 -HT 抗血清分别与变异株和非变异株H抗原进行试管凝集试验 ,均为强凝集。两种菌株O15 7和H7特异性基因均为阳性。变异株与非变异株生化特性基本相同 ,但变异株不发酵棉子糖。两种菌株在麦康凯及山梨醇麦康凯培养基上菌落形态特征没有区别。在ChromagarO15 7显色培养基上培养 4 8h ,变异株为浅紫红色 ,菌落周边呈毛边状 ,非变异株为紫红色 ,菌落边缘整齐。变异株在用棉子糖代替乳糖制备的棉子糖麦康凯培养基上菌落呈粉红色 ,非变异株呈红色。菌落计数显示变异率为 99 80 %。结论　CVa9菌株在保存过程中O抗原发生变异 ,菌落形态及部分生化特征亦有所改变。变异株和非变异株均携带O15     

免疫球蛋白对体外生长的几株恶性疟原虫的不同作用

作者测试了从巴布亚-新几内亚疟区居民血清中分离出的免疫球蛋白对恶性疟原虫体外生长的作用,检测了对有抑制作用的免疫球蛋白敏感性不同的虫株的蛋白质和抗原成分。试验的恶性疟原虫共4株,其中3株(FCQ2,FCQ27,FCQ46)取自巴布亚-新几内亚,在实验室保种培养2年以上。1株(KI)取自泰国。保种系用HEPES缓冲的RPMI 1640培养基加10%人血清,按照Trager和Jensen蜡罐法培养。     

在特定的无血清条件下的弓形虫体外培养

迄今为止,用作弓形虫体外培养的培养基为保证虫体与宿主细胞的生长总是富含血清或其他成份。但发现培养基添加血清能影响其细胞内弓形虫滋养体的释出及其活力。为解决这一问题,作者用特定组成的培养基完全地取代富含血清培养基,予以培养条件     

一株阴道毛滴虫释放的一种RNA病毒及其细胞病理学

近年有些作者对不同原虫中存在的小RNA病毒曾作描述。本文作者研究了一株1964年分离并传代的阴道毛滴虫(123414)及另两株不同来源的阴道毛滴虫(虫株87464和30001)。细胞按常规置无血清的C13培养基(pH 7)34℃培养。对照虫株也置于C13和Diamond′s培养基,pH6,34℃或37℃下培养。在培养3—7天后,显微镜下观察各株细胞贴壁情况。离心收集阴道毛滴虫,     

血清诊断美洲利什曼病的间接萤光抗体试验

抗原:在感染人体巴西利什曼原虫原鞭毛体的非洲绿猴的单层肾脏Vero细胞中生长的原虫,经用15%兔血琼脂斜面培养使其大量繁殖后,浮悬在Eagle基础保种培养基中,并接种于组织培养,在35℃孵箱内培育6天后,倾去保种培养基以去掉细胞外的原虫,将感染原虫的细胞放入佛罗拿缓冲盐水(VBS)中,用0.02%EDTA处理,使感染原虫的细胞破裂,释出细胞里的无鞭毛原虫     

感染疟原虫的红细胞突起物形成的逆转

人恶性疟原虫株在培养基中培养可发生某些改变,有突起株(K~+)经长期培养可变为无突起株(K~-)。为了鉴定体外培养是否会引起K~-变种或仅是亚群生长,作者将K~+进行克隆,然后筛选培养K~-细胞,对K~-细胞中的原虫进行再克隆,选出K~+细胞。最终证明在K~+存在一种蛋白质,而K~-是没有的。     

卡氏肺孢子虫体外无细胞培养的建立

目的建立卡氏肺孢子虫(Pneumocystis carinli，Pc)的体外无细胞系培养体系。方法建立Wistar大鼠卡氏肺孢子虫肺炎模型，Pereoll不连续密度梯度离心方法分离、纯化卡氏肺孢子虫。用添加s-腺苷甲硫氨酸(SAM)、N-乙酰氨基葡萄糖苷、腐胺等辅助剂的IMDM培养基体外培养虫体，用四胺银染色和姬姆萨染色方法计数虫体生长增殖情况，并以透射电镜观察培养虫体的超微结构形态。结果添加s-腺苷甲硫氨酸(SAM)等辅助剂的IMDM培养基(在37℃，pH8．0，5％CO2、95％O2条件)能支持Pc的体外生长。Pc包囊原代培养10天可增殖8～9倍，滋养体可增殖11～12倍。原代与传代培养、新鲜分离的与冻存复苏的虫体在培养基内的生长增殖有差别。结论建立了一个比较稳定的Pc体外无细胞系培养体系。     

Intestinal epithelial culture under an air‐liquid interface: a tool for studying human and mouse esophagi

This study investigated whether an intestinal epithelial culture method can be applied to mouse and human esophageal cultures. The esophagi harvested from 1‐day‐old mice and adult humans were maintained in collagen gels. A commercially available culture medium for human embryonic stem cells was used for the human esophageal culture. We discovered that the intestinal epithelial culture method can be successfully applied to both mouse and human esophageal cultures. The long‐term cultured esophageal organoids were rod‐like luminal structures lined with myofibroblasts. We discovered that regeneration of the esophageal mucosal surface can be almost completely achieved in vitro, and the advantage of this method is that organoid cultures may be generated using host‐derived fibroblasts as a niche. This method is a promising tool for mouse and human research in intestinal biology, carcinogenesis, and regenerative medicine.     

刚地弓形虫细胞培养的研究Ⅱ．RH株速殖子细胞培养上清对成囊株速殖子侵入细胞的影响

本文以ＲＨ株细胞培养上清（ＳＲＣ）替代ＰＬＡ２加入培养系统，对三株成囊株４次培养及与含ＰＬＡ２对照培养的观察表明，含ＳＲＣ的培养系统较常规培养检出时间提前，阳性率提高，在本文条件下与含ＰＬＡ２４０Ｕ者效果相当，提示ＳＲＣ可作为ｐＬＡ２的廉价实用的替代物用于弓形虫病的病原检测。     

胆石症并胆道感染患者胆汁与血细菌培养比较及药敏分析

目的探讨胆石症并胆道感染患者胆汁与血中细菌分布、细菌培养阳性率及二者关系,并探讨菌群对抗生素的敏感性,以指导临床用药。方法对浙江省象山县第一人民医院2003-01~2004-05胆总管结石并胆道感染患者120例通过ERCP胆管深插管抽取胆汁进行细菌培养加药敏试验,同时取40例进行血培养。结果120例胆汁培养67例检出细菌,阳性率为55.8%。40例血培养10例检出细菌,阳性率为25.0%。均主要由G-菌中的大肠埃希菌属、克雷伯杆菌属和肠杆菌属构成。10种抗生素敏感谱中,亚胺培南、美洛匹宁、阿米卡星、庆大霉素等抗菌效果优于氨苄青霉素、替卡西林、头孢噻吩、头孢西丁、头孢呋辛等(P<0.05)。结论胆石症并胆道感染患者胆汁及血中细菌分布均以G-菌为主,且在菌属构成上差异无显著性。但胆汁中细菌阳性率明显高于血培养中细菌阳性率(P<0.01)。治疗上宜据药敏试验主要针对G-杆菌应用敏感抗生素。     

组织贴块法人的支气管上皮细胞的原代培养

目的 通过组织贴块法建立人的支气管上皮细胞原代培养的方法.方法 采用无血清支气管上皮培养基,对经手术切除获得的支气管进行组织贴块法培养获得的细胞进行原代培养和传代,通过倒置显微镜以及细胞免疫化学观察和鉴定细胞.结果 此方法获得的细胞成活率高、纯度高,细胞呈扁平、多边形,象铺路的鹅卵石样分布.细胞角蛋白表达阳性.结论 组织贴块法是一种简便、有效的培养人的支气管上皮细胞的方法,无血清培养基可提供满意的生长条件,提高上皮细胞的纯度,为进一步研究不同的呼吸系统疾病提供了良好的模型.     

兔软骨细胞培养方法研究

Manning 和 Bonner1967年首先把胰蛋白酶和细菌胶原酶结合使用消化关节软骨基质获得大量纯度高的软骨细胞,此后,软骨细胞培养研究工作得到很大发展并取得丰硕成果。现这一方法经改进后在国外已被广泛使用,在我国尚未见有关这方面报导。本文以体外单层兔软骨细胞培养为例,简要介绍用透明质酸酶、胰蛋白酶和胶原酶消化分离兔关节软骨细胞的实验材料准备,操作方法步骤,并对实验结果作了简要讨论。希望通过这一方法介绍,对从事软骨疾病研究、大骨节病病因研究人员有所助益。     

Revisiting factors associated with blood culture positivity: Critical factors after the introduction of automated continuous monitoring blood culture systems

Blood culture is the main tool used to identify causative pathogens. Adequate volume and number of culture sets are considered key to blood culture positivity rate. It is not known whether these factors remain critical to the positivity rate after the introduction of automated continuous blood culture system monitoring.We measured blood volume per bottle and described the distribution of blood volume and number of culture sets. Multivariate logistic regression was performed to determine the independent association of blood volume, number of culture sets, diagnosis of sepsis in a patient, and other covariates with blood culture results.Only 6.9% of the blood culture bottle volumes complied with the guidance (8–10 mL), with the highest culture positivity rate (18%). Of the culture events, only one set of blood was cultured in 60.9% of events. In the multivariate analysis, blood culture volume per event (odds ratio [OR], 1.09 [95% confidence interval [CI], 1.06–1.11]), patients with a diagnosis of sepsis (OR, 2.86 [95% CI, 2.06–3.98]), and samples from the emergency department (OR, 2.29 [95% CI, 1.72–3.04]), but not the number of culture sets (OR, 0.74 [95% CI, 0.50–1.12]), were observed to be statistically significant with respect to blood culture positivity rate.Our results revealed that the total blood culture volume and the diagnosis of sepsis were critical factors affecting blood culture positivity rate. However, the proportion of blood culture bottles with the optimal blood volume was very low, and optimizing blood volume would be key to increasing blood culture positivity rate.     

Stimulation of autoantibody production in normal blood lymphocytes by malaria culture supernatants

Supernatants from Plasmodium falciparum cultures containing soluble parasite material were mitogenic for normal human peripheral blood mononuclear cells (MNC) in vitro. This was evidenced by blast transformation and significant incorporation of 3H-thymidine and confirms earlier reports of the mitogenic potential of malaria parasites. Lymphocyte activation by these malaria derived products was polyclonal as demonstrated by increased secretion of IgA, IgG and IgM by the stimulated cells. Using rat tissues and Hep-2 cells as substrates, autoantibody activity was found in the IgM fraction of the secreted immunoglobulin. Speckled anti-nuclear (ANA) antibody, anti-globulins (rheumatoid factor) and anti-intermediate filament antibodies were produced by the stimulated lymphocytes. No significant immunoglobulin secretion with autoantibody specificity was found in control cultures in which normal MNC were incubated with supernatants from non-parasitized red cell cultures. The data supports the suggestion that polyclonal lymphocyte activation by parasite derived products occurs in vivo and, in addition, provides an explanation for the presence of autoantibodies in the serum of malaria patients.     

Predicting consumer behavior with Web search

Recent work has demonstrated that Web search volume can “predict the present,” meaning that it can be used to accurately track outcomes such as unemployment levels, auto and home sales, and disease prevalence in near real time. Here we show that what consumers are searching for online can also predict their collective future behavior days or even weeks in advance. Specifically we use search query volume to forecast the opening weekend box-office revenue for feature films, first-month sales of video games, and the rank of songs on the Billboard Hot 100 chart, finding in all cases that search counts are highly predictive of future outcomes. We also find that search counts generally boost the performance of baseline models fit on other publicly available data, where the boost varies from modest to dramatic, depending on the application in question. Finally, we reexamine previous work on tracking flu trends and show that, perhaps surprisingly, the utility of search data relative to a simple autoregressive model is modest. We conclude that in the absence of other data sources, or where small improvements in predictive performance are material, search queries provide a useful guide to the near future.