利用ELISA技术评估尘螨变应原的稳定性

目的检测尘螨变应原的稳定性。方法利用ELISA技术检测在37℃处理后尘螨变应原的稳定性。结果尘螨变应原溶液在37℃处理6d后,利用人抗血清可以检测到其活性下降,而使用兔抗血清时尘螨抗原生物活性的变化在37℃处理45d后仍未检测到。结论尘螨变应原的稳定性应利用IgE抗体来检测,表明尘螨变应原的生物活性在37℃处理45d的过程中逐渐下降。     

基于文字音韵的弧蜮病解读

《金匮要略》中“狐惑”两字从传统考据之音韵、训诂探源当为“弧蜮”。《说文解字注》指出“狐”并非指狐疑之意,当为“弧”之错讹,为“蜮”的外形特征,指身体呈弧形;“惑”非为疑惑之意,当读“蜮”,即江淮水滨之虫。马王堆出土文献《疗射工毒方》对“蜮”的记载突出其具有“射”的功能,符合“蜮”身体呈弧形的外形特征。“狐惑”表现出的“状如伤寒”“蚀于喉”“蚀于阴”“面目乍赤乍黑乍白”等症状与恙虫致病之“偶有畏寒,伴有剧烈头痛全身疼痛乏力,食欲不振;腋下、会阴、肛门出现无痛的庁;丘疹眼结膜的充血”等表现十分吻合。针对此“弧蜮”症状与恙虫致病,通过地下出土与现存传世文献“二重证据”相互佐证,运用训诂学方法,推理出“狐”为“弧”之错讹,“惑”与“蜮”同源,明晰“狐惑”之源流。     

屋尘螨特异性免疫治疗的哮喘患儿对尘螨过敏原免疫反应的变化

目的观察经屋尘螨特异性免疫治疗(SIT)的哮喘患儿对尘螨过敏原特异性免疫反应的变化。方法选取54例尘螨过敏性哮喘患儿,其中男性38例,女性16例;年龄6~11岁,平均年龄9岁1个月。27例经标准化屋尘螨过敏原皮下SIT疗程1.5~2.0年患儿作为SIT组,另27例未经SIT患儿作为对照组。应用最大呼气流量-容积曲线测定方法测定肺通气功能,参数为第1秒用力呼气容积(FEV1)占预计值百分比(FEV1%);应用荧光酶联免疫法测定2组患儿血清屋尘螨特异性IgE和特异性IgG4浓度;分离患儿外周血单个核细胞(PBMC),经质量浓度20μg/mL屋尘螨蛋白浸液刺激并培养48 h后,检测CD4+CD25+Foxp3+调节性T淋巴细胞(Treg细胞)的相对数量和可分泌白细胞介素10(IL-10)的Treg细胞百分比;同时留取细胞培养上清液应用酶联免疫吸附分析检测细胞因子IL-4、干扰素-γ(IFN-γ)和转化生长因子β1(TGF-β1)水平。结果 SIT组患儿肺功能FEV1%与对照组相比,差异无统计学意义(P>0.05);SIT组IgG4水平显著高于对照组,差异有极显著统计学意义(P<0.000 1)。SIT组IgE水平与对照组相比,差异无统计学意义(P>0.05);两组刺激后Treg细胞百分数分别与各自该组内未经刺激时基础水平的差异无统计学意义。两组间PBMC无论是否经屋尘螨蛋白浸液刺激时Treg细胞百分数的差异无统计学意义(P>0.05);SIT组刺激后Treg细胞百分数高于自身未经刺激基础水平1.56倍。对照组经刺激后与未接受刺激比较,Treg细胞百分数无显著变化;SIT组PBMC培养上清液中IL-4和TGF-β1浓度显著低于对照组,IFN-γ浓度显著高于对照组,差异有统计学意义(P<0.05)。结论屋尘螨SIT的哮喘患儿对尘螨过敏原特异性免疫反应表现为Treg细胞比率增加,Th1/Th2免疫反应平衡改变,屋尘螨特异性IgG4浓度增高,提示发生对尘螨过敏原特异性免疫耐受的特征。     

Mast cells derived from human induced pluripotent stem cells are useful for allergen tests

Background

Several methods have been developed to detect allergen-specific IgE in sera. The passive IgE sensitization assay using human IgE receptor-expressing rat cell line RBL-2H3 is a powerful tool to detect biologically active allergen-specific IgE in serum samples. However, one disadvantage is that RBL-2H3 cells are vulnerable to high concentrations of human sera. Only a few human cultured cell lines are easily applicable to the passive IgE sensitization assay. However, the use of human induced pluripotent stem cells (iPSCs) to generate human mast cells (MCs) has not yet been reported.

尘螨肠道微生物蛋白Hypothetical protein CE2118的表达及纯化

目的 研究尘螨肠道微生物蛋白Hypothetical protein CE2118的表达和纯化,为研究其在尘螨疫苗免疫治疗中的作用奠定基础。方法 采用生物信息学方法,根据GenBank中Hypothetical protein CE2118蛋白的基因序列,将其中稀有密码子改造为大肠杆菌常用密码子并进行二级结构优化,合成Hypothetical protein CE2118基因,构建原核表达载体pGEX 6P-1-hypothetical protein CE2118并经酶切鉴定,在大肠埃希菌Rosetta(DE3)中用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,重组产物采用GST亲和层析纯化柱纯化。结果 经密码子改造和二级结构优化后Hypothetical protein CE2118基因长度为588bp,其编码蛋白理论分子量为21 kDa。重组表达载体经酶切鉴定与理论推测结果相符,该基因经IPTG诱导在大肠埃希菌Rosetta(DE3)中得到高效的可溶性表达,纯化后的重组蛋白分子量约为21 kDa,其单一蛋白纯度达95%以上。结论 本研究成功构建了Hypothetical protein CE2118基因的pGEX6P-1原核重组质粒,获得的可溶性重组蛋白为进一步研究肠道微生物蛋白Hypothetical protein CE2118在尘螨疫苗免疫治中的作用机理奠定基础。     

Is asthma a protective factor for dengue fever? In vitro experiment and nationwide population-based cohort analysis

BackgroundDengue fever (DF) is the most rapidly spreading mosquito-borne viral disease. Practical vaccines or specific therapeutics are still expected. Environmental factors and genetic factors affect the susceptibility of Dengue virus (DV) infection. Asthma is a common allergic disease, with house dust mites (HDMs) being the most important allergens. Asthmatic patients are susceptible to several microorganism infections.MethodsA nationwide population-based cohort analysis was designed to assess whether to determine whether asthma can be a risk factor for DF.ResultsUnexpectedly, our data from a nationwide population-based cohort revealed asthmatic patients are at a decreased risk of DF. Compared to patients without asthma, the hazard ratio (HR) for DF in patients with asthma was 0.166 (95% CI: 0.118–0.233) after adjustment for possible confounding factors. In the age stratification, the adjusted HR for DF in young adult patients with asthma was 0.063. Dendritic cell-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN) of dendritic cells (DCs) is an important entry for DV. Through another in vitro experiment, we found that HDM can diminish surface expression of DC-SIGN in monocyte-derived DCs and further decrease the cellular entry of DV.ConclusionsDecreased DC-SIGN expression in DCs of allergic asthmatic patient may be one of many factors for them to be protected against DF. This could implicate the potential for DC-SIGN modulation as a candidate target for designing therapeutic strategies for DF.     

蠕形螨自然死亡的判断方法探讨

目的对人体蠕形螨的自然死亡判断方法进行探讨和分析。方法用透明胶纸法取活螨,于Version10.0金盘多媒体摄影系统观察下,每隔3h分别观察并拍摄虫体死亡的过程,记录虫体形态、结构、颜色和内含物颗粒变化。结果活螨虫体浅黄色、半透明,左右对称,形态规则,结构完整清晰,末体中部聚集了清晰的深色颗粒,细小均匀,环行纹理清晰、规则。运动活泼,口器、四肢及末体摆动明显,升温后活动加剧。死螨形态明显改变,虫体肿胀,表膜纹理模糊或消失,体周出现油滴状分泌物堆积,发生皱缩变形、边界不清,破裂后内容物外溢等。结论此法有助在普通光学显微镜下直接、快速、准确判断虫体死亡,为深入研究奠定基础。