Fast Dipstick Dye Immunoassay for Detection of Immunoglobulin G (IgG) and IgM Antibodies of Human Toxoplasmosis

A dipstick dye immunoassay (DDIA) was developed to detect immunoglobulin G (IgG) or IgM antibodies of toxoplasmosis infection in humans. The assays employ a blue colloidal dye particles (D-1) conjugated to sheep anti-human IgG and rabbit anti-human IgM as the visualizing agents and a soluble antigen of tachyzoites of Toxoplasma gondii strain RH (TSA) as the detective antigen. The mixture of dye-labeled anti-human antibody-special human antibody was captured by TSA onto a nitrocellulose membrane dipstick by means of immunochromatography. The assays are rapid (the whole test can be completed within 15 min), simple, and cheap, and they don't require any equipment. They are sensitive and specific for the detection of anti-Toxoplasma IgG or IgM antibodies and generally agree closely with the results from the enzyme-linked immunosorbent assay. The assays are especially suitable for field applications.     

The Distribution of Synaptotagmin II in RBL-2H3 and Its Regulation on Exocytosis of Lysosomes in RBL-2H3

Synaptotagmin(Syt)constitutes a family of membrane-trafficking proteins,so far nearly 20 Syts have beendiscovered.Extensive work showed that synatotagmins were a potential Ca~(2+) sensor for regulated exocytosis.Thisstudy was to investigate the expression and location of synaptotagmin Ⅱ(Syt2)in RBL-2H3(RBL)and its role inregulating exocytosis of RBL.The expression of Syt2 in RBL was confirmed by Western blot.The recombinantexpression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation,the stabletransfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm.To analyze the role of Syt2 during exocytosis of RBL,therelease of cathepsin D was assayed by immunoblotting.Compared with control,the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis oflysosomes in RBL.Cellular & Molecular Immunology.2005;2(3):205-209.     

CD3ζ链基因在脐带血T细胞及其CD4+和CD8+T细胞亚群中的表达特点

目的 了解脐带血T细胞及其CD4 和CD8 T细胞亚群的信号传导分子CD3ζ基因的表达特点.方法 采用SYBR Green Ⅰ荧光定量PCR和相对定量分析法检测60例脐带血单个核细胞和12例纯化脐带血CD4 及CD8 T细胞的CD3ζ基因的表达情况,以β2微球蛋白基因(β2M)作为内参,采用相对定量公式:2-△Ct×100%,计算CD3ζ基因相对mRNA表达量,60例健康成人作为对照.并根据荧光定量PCR熔解曲线特点和序列分析了解CD3ζ基因突变情况.结果 全部脐带血和健康成人外周血单个核细胞均表达CD3ζ基因,不同个体CD3ζ基因表达量有所差异,脐带血T细胞CD3ζ基因相对mRNA表达量为6.7%±5.56%,CD4 和CD8 T细胞的CD3ζ基因相对mRNA表达量分别为6.74%±2.0%和6.88%±1.76%,三者CD3ζ基因表达量均明显高于健康成人(P=0.000,P=0.034,P=0.000).序列分析结果显示尚未发现国外文献所报道的ζ链剪接异构体.结论 本研究率先报道了脐带血T细胞及其CD4 和CD8 T细胞亚群的CD3ζ基因mRNA的表达水平,为进一步了解脐带血T细胞免疫学特点提供新的基础资料.     

基于遗传算法的经颅磁刺激线圈阵列设计与场分布计算

为解决经颅磁刺激的兴奋点定位问题提出了以圆环面聚焦线圈阵列实现电磁聚焦的方案。而后,使用经过改进的自适应遗传算法对注入阵列各单元线圈电流的大小和相位进行了优化,在目标区域内很好的实现了电磁聚焦。通过计算,显示了线圈阵列在优化后的电流组态下产生的磁场和电场在计算区域内的分布和二维等高线图,表明了该聚焦线圈阵列可在目标区域产生具有良好聚焦性能的电场和磁场分布,并具有可同时聚焦至多个目标的能力。     

RGS1对RAW264.7细胞共刺激分子和细胞因子表达的调节

目的:研究RGS1基因对细胞表面分子表达以及细胞因子分泌的影响.方法:通过克隆RGS1基因、转染RAW264.7细胞、建立稳定转染的细胞系, 利用流式细胞术检测, 基因芯片分析初步探讨RGS1基因的功能.结果:RGS1基因能够抑制NF-κB转录因子活性; RGS1基因明显增加RAW264.7细胞表面CD86分子的表达, 降低CD80表达; 在不同Toll样受体配体刺激后, CD40、 CD80和PD1水平低于对照组.同时, RGS1基因使IL-6、IL-15的表达明显升高, IL-10、 IL-1的表达明显降低.结论:RGS1基因可调节免疫相关细胞表面分子和细胞因子的分泌, 影响Toll 样受体信号通路,表明RGS1可能在免疫调节方面起重要作用.     

逆转录病毒载体介导乙型肝炎病毒反义基因的转录表达

为了探索在真核细胞内转录表达乙型肝炎病毒（ＨＢＶ）反义核酸的方法，用基因重组技术将ＨＢＶ前Ｃ／Ｃ基因（ＰｒｅＣ／Ｃ）和前Ｓ／Ｓ基因（ＰｒｅＳ／Ｓ）片段反向插入逆转录病毒载体质粒，再将重组体分别转染ＰＡ３１７包装细胞，进而获得能够介导ＨＢＶ反义基因向小鼠ＮＩＨ３Ｔ３细胞转移表达的重组逆转录病毒。经分子杂交试验表明，含有ＨＢＶ反义基因的重组逆转录病毒序列已经整合到转染的ＰＡ３１７细胞染色体上；转导的ＮＩＨ３Ｔ３细胞内有ＨＢＶ反义ＲＮＡ转录表达。结论：逆转录病毒载体包装细胞系统能够介导ＨＢＶ反义基因在真核细胞中转录表达，因而有可能利用反义技术和基因转移方法进行抗－ＨＢＶ基因治疗     

In vitro dedifferentiation of fetal porcine pancreatic tissue prior to transplantation as islet-like cell clusters

The fetal porcine pancreas under experimental conditions can be transplanted in the form of explants or islet-like cell clusters (ICCs) to normalize blood glucose levels in diabetic recipients. ICCs are released from the collagenase-digested pancreas and require a 4- to 5-day culture period for their complete formation. In order to maximize insulin producing beta cell differentiation following transplantation, an understanding of ICC development is essential to utilize this alternative treatment for type 1 diabetes. In this study a role is proposed for exocrine cells in the generation of the multipotent pancreatic precursor cells during the culture period. Acinar cells undergo dedifferentiation during the initial stages of the culture period into multipotent pancreatic precursor cells, previously called protodifferentiated cells. The progressive loss of exocrine differentiation appears to involve rapid degranulation of zymogen granules by exocytosis and loss of the prominent secretory apparatus. These processes occur in parallel with a significant reduction in the expression of lipase in the period from day 0 to day 5 and simultaneously there is an increase in the epithelioid/ductal cell marker, cytokeratin 20. Using proliferating cell nuclear antigen, cell proliferation during the culture period does not appear to account for the increase in epithelioid/ductal cells. Further the rates of apoptosis and necrosis which were identified using the TUNEL technique and propidium iodide, respectively, do not appear to account for the reduction in exocrine cell numbers. Exocrine cell dedifferentiation appears to increase the pool of protodifferentiated cells which have the potential to develop into the insulin-producing beta-cell population following transplantation into the diabetic recipient     

Afferent innervation of the utricular macula in pigeons

Biotinylated dextran amine (BDA) was used to retrogradely label afferents innervating the utricular macula in adult pigeons. The pigeon utriclar macula consists of a large rectangular-shaped neuroepithelium with a dorsally curved anterior edge and an extended medioposterior tail. The macula could be demarcated into several regions based on cytoarchitectural differences. The striola occupied 30% of the macula and contained a large density of type I hair cells with fewer type II hair cells. Medial and lateral extrastriola zones were located outside the striola and contained only type II hair cells. A six- to eight-cell-wide band of type II hair cells existed near the center of the striola. The reversal line marked by the morphological polarization of hair cells coursed throughout the epithelium, near the peripheral margin, and through the center of the type II band. Calyx afferents innervated type I hair cells with calyceal terminals that contained between 2 and 15 receptor cells. Calyx afferents were located only in the striola region, exclusive of the type II band, had small total fiber innervation areas and low innervation densities. Dimorph afferents innervated both type I and type II hair cells with calyceal and bouton terminals and were primarily located in the striola region. Dimorph afferents had smaller calyceal terminals with few type I hair cells, extended fiber branches with bouton terminals and larger innervation areas. Bouton afferents innervated only type II hair cells in the extrastriola and type II band regions. Bouton afferents innervating the type II band had smaller terminal fields with fewer bouton terminals and smaller innervation areas than fibers located in the extrastriolar zones. Bouton afferents had the most bouton terminals on the longest fibers, the largest innervation areas with the highest innervation densities of all afferents. Among all afferents, smaller terminal innervation fields were observed in the striola and large fields were located in the extrastriola. The cellular organization and innervation patterns of the utricular maculae in birds appear to represent an organ in adaptive evolution, different from that observed for amphibians or mammals.     

先天性生殖器官异常孕妇的围生结局分析

目的探讨先天性生殖器官异常孕妇的围生结局。方法选择2009年1月至2019年12月,于四川大学华西第二医院活产分娩的99779例单胎妊娠孕妇为研究对象。根据是否合并先天性生殖器官异常,将其分为观察组(n=324,合并)及对照组(n=99455,未合并)。回顾性分析2组孕妇的临床病例资料,总结观察组不同类型先天性生殖器官异常孕妇的围生结局特点。采用χ^(2)检验及独立样本t检验,对2组孕妇妊娠期各并发症发生率、剖宫产率及新生儿出生体重等,进行统计学分析。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求。结果①观察组孕妇最常见先天性生殖器官异常前3位依次为纵隔子宫、单角/残角子宫及弓形子宫,分别占35.2%(114/324)、26.2%(85/324)及13.9%(45/324)。包括既往妊娠次数在内,观察组孕妇共计妊娠571次,其中弓形子宫、纵隔子宫孕妇的自然流产率位列前2位,分别为38.5%(35/91)、28.6%(61/213);阴道畸形孕妇足月产率(70.0%),高于子宫畸形者(52.8%),并且差异有统计学意义(χ^(2)=4.432、P=0.035)。②观察组孕妇早产、胎膜早破、胎位异常、胎盘早剥、胎儿生长受限(FGR)、胎盘黏连/植入、子宫破裂及剖宫产率分别为29.6%、29.3%、36.1%、5.9%、2.5%、21.0%、3.4%、78.7%,均分别高于对照组孕妇的9.8%、22.3%、5.7%、1.2%、1.0%、11.5%、1.5%、62.0%;而羊水过少率(0.9%)及新生儿出生体重[(2913±652)g],则显著低于对照组的2.9%、(3254±445)g,并且上述差异均有统计学意义(均为P<0.05)。2组孕妇前置胎盘、妊娠期高血压疾病(HDCP)、产后出血(PPH)及子宫切除率比较,差异均无统计学意义(P>0.05)。结论先天性生殖器官异常孕妇的母、胎围生期并发症发生风险高。提高该病孕妇的孕前及早孕期诊断率,加强围生期保健及处理,可改善其围生结局。