Regular microarray of Au flower-like structure formed by template-assisted electrodeposition: Influence factors and their electrochemical behavior

This paper describes a template-assisted method that combines the advantages of photolithography and electrochemical synthesis to construct a wafer-scale Au flower-like structure microarray. The detailed information about the effect of concentration of electrolyte, pattern of template substrate, applied potential, and surfactant on the formation of Au flower-like microarray is investigated. The possibility of using such a microarray as electrodes for their electrochemical properties is also tested by CV experiments.     

Representation of Women in News and Photos: Comparing Content to Perceptions

This study uses a feminist framework of masculine cultural hegemony to examine the representation of women in two newspapers—a medium-sized newspaper (Study 1) and a larger newspaper (Study 2). Surveys gauged news staff and news reader perceptions of female representation in news content to determine if perceptions matched content-analysis findings. Content-analysis results revealed a greater proportion of males than females in both content and photos for both studies. Female staffers were more likely than their male counterparts to perceive this disparity, and news readers perceived an even greater disparity than the news staff. News staffs were less likely to perceive disparities in the editorial, travel, and entertainment sections. We discuss the findings in relation to cultural hegemony and the representation of women in the news.     

Identification and Functional Analysis of MicroRNAs in Mice following Focal Cerebral Ischemia Injury

Numerous studies have demonstrated that genes, RNAs, and proteins are involved in the occurrence and development of stroke. In addition, previous studies concluded that microRNAs (miRNAs or miRs) are closely related to the pathological process of ischemic and hypoxic disease. Therefore, the aims of this study were to quantify the altered expression levels of miRNAs in the infarct region 6 h after middle cerebral artery occlusion (MCAO)-induced focal cerebral ischemia in mice using a large-scale miRNAs microarray. Firstly, MCAO-induced cerebral ischemic injuries were investigated by observing the changes of neurological deficits, infarct volume and edema ratio. One hundred and eighteen differentially expressed miRNAs were identified in the infarct region of mice following the MCAOs compared with sham group (p < 0.05 was considered as significant). Among these 118 significantly expressed microRNAs, we found that 12 miRNAs were up-regulated with fold changes lager than two, and 18 miRNAs were down-regulated with fold changes less than 0.5 in the infarct region of mice following the 6 h MCAOs, compared with the sham group. Then, these 30 miRNAs with expression in fold change larger than two or less than 0.5 was predicted, and the functions of the target genes of 30 miRNAs were analyzed using a bioinformatics method. Finally, the miRNA-gene network was established and the functional miRNA-mRNA pairs were identified, which provided insight into the roles of the specific miRNAs that regulated specified genes in the ischemic injuries. The miRNAs identified in this study may represent effective therapeutic targets for stroke, and further study of the role of these targets may increase our understanding of the mechanisms underlying ischemic injuries.     

质粒剂量和免疫次数对基因枪免疫DNA疫苗免疫反应的影响

目的研究质粒剂量和免疫次数对基因枪免疫DNA疫苗免疫反应的影响。方法构建携带HIV-1 Env gp145基因的DNA疫苗质粒p1.0-Env,分别采用肌肉注射和基因枪方法免疫BALB/c小鼠。其中,基因枪免疫采用不同剂量或不同次数。ELISA法检测小鼠体内Env特异性抗体水平。结果基因枪免疫小鼠诱导的抗体水平显著高于肌肉注射免疫。在0.1～2.25μg剂量范围内,基因枪免疫小鼠体内的Env特异性抗体水平随疫苗剂量的增加不断提高。加大免疫剂量至5μg和10μg,不能提高体液免疫反应水平。基因枪免疫1次和2次所诱导的特异性抗体水平很弱,免疫3次可诱导高水平的抗体应答。结论基因枪免疫能有效提高DNA疫苗所诱导的抗体应答,小鼠的最适免疫剂量约为2.25μg,2.25μg免疫3次可诱导较强的体液免疫反应。     

从噬菌体表面展示肽库中筛选边缘无浆体膜表面蛋白5单克隆抗体识别的抗原表位

目的从噬菌体表面展示肽库中筛选边缘无浆体膜表面蛋白5(Membrane surface protein5,MSP5)单克隆抗体识别的抗原表位。方法用MSP5单克隆抗体1D8作为靶标,对噬菌体展示随机12肽库进行筛选,通过ELISA和竞争抑制ELISA鉴定筛选克隆的结合特性,并提取阳性克隆的单链DNA,进行测序分析。结果从表面展示随机肽序列的噬菌体文库中筛选到与MSP5单抗1D8特异结合的噬菌体克隆,其一致序列为LING。竞争抑制试验表明,含特异序列的克隆能与MSP5重组蛋白抗原竞争。结论初步确定MSP5单克隆抗体1D8的抗原表位为线性表位,为进一步研究其在边缘无浆体诊断及新型疫苗研制中的作用奠定了基础。     

Fluctuation of Global Gene Expression by Endogenous miRNA Response to the Introduction of an Exogenous miRNA

Most of the intracellular endogenous microRNAs (endo-miRNAs) are considered to be saturated in Argonaute (Ago) proteins in the RNA-induced silencing complexes (RISCs). When exogenous miRNAs (exo-miRNAs) are introduced into cells, endo-miRNAs in the RISC may be replaced with exo-miRNAs or exo-miRNAs, and endo-miRNAs might also compete for the position in the newly synthesized RISC with each other. This would lead to the fluctuation of global gene expression not only by repression of exo-miRNA target gene expression, but also by the increase of the endo-miRNA target gene expression. In the present study, we quantified the changes in the expression levels of target genes of exo-miRNA and endo-miRNA in the cells transfected with fifteen different exo-miRNAs by microarray experiments. Different exo-miRNAs increased ratios of expression levels of target genes of a given endo-miRNA to different extents, suggesting that the replacement efficiencies might differ according to the exo-miRNA types. However, the increased ratios in the expression levels of each endo-miRNA target genes by the transfection of any particular exo-miRNA were mostly equivalent, suggesting that the endo-miRNAs present in the RISC might be replaced with excessive exo-miRNAs at similar levels, probably because they exist in single-stranded forms in the RISC.     

从噬菌体抗体库中筛选出抗原结合活性相近、表达水平不同的单链抗体

从抗人膀胱癌改形噬菌体抗体库中筛选出一组相对亲和力常数和细胞结合活性相近的单链抗体，而它们的结合活性均明显高于起始的鼠源单链抗体。在亲和力淘选的过程中，这一组抗体的表达量存在显著差异，这在一定程度上影响到筛选过程中ELISA鉴定的结果，使整个筛选过程并不仅仅是对高亲和力抗体的筛选，也同时得到了高表达量的抗体。     

血样中血小板污染对凝聚胺抗体筛检和交叉配血的影响

目的探讨血小板污染对凝聚胺抗体筛检和交叉配血试验的干扰。方法应用自然沉降和离心方法去除血液标本中的血小板,用凝聚胺方法进行血小板去除前后抗体筛检和交叉配血试验。瑞氏染色显微镜下观察血小板聚集。结果自然沉降的标本有大量的絮状团块,镜下可见血小板聚集,而非红细胞凝块。不同离心力去除血小板污染的程度不同,离心力越大,血小板聚集越少,离心力达2200g,可完全去除血小板污染,对凝聚胺抗体筛检和交叉配血试验的干扰消失。结论血浆标本中血小板污染可对凝聚胺抗体筛检和交叉配血试验产生干扰,但可以采用离心法去除。     

miR-140-5P 参与调控颈椎软骨终板退变

目的：探讨表达谱芯片初步分析 microRNA差异表达是否参与调控颈椎软骨终板退变。方法选择25例颈椎软骨终板退变、需要行手术治疗的患者为病例组,15例无椎间盘软骨终板退变患者为对照组。选取病例组2例和对照组2例标本进行microRNA芯片差异表达分析。然后采用miRCURYTM LNA 18．0表达谱芯片进行原位杂交,由 Axon GenePix 4000B microarray扫描仪采集芯片荧光值。芯片结果经病例组15例和对照组15例标本进行 Real-time PCR和Western blot验证。结果病例组发现22个上调和19个下调 microRNA。Real-time PCR验证：病例组miR-140-5P表达量低于对照组的3倍、miR-140-5P的靶基因 Adamts-5和 Dnpep的表达量分别高于对照组的3倍和3．5倍（均P＜0．05）。miR-140-5P 与 Adamts-5和 Dnpep 的 miRNA表达呈负相关（r＝-0．84、-0．76,均P＜0．05）。Western blot结果示：病例组软骨终板 Adamts-5和 Dnpep的蛋白表达分别为0．66±0．46和0．57±0．32,均高于对照组软骨终板的0．22±0．06和0．26±0．08（均P＜0．05）。结论本研究首次报道了 miR-140-5P参与调控颈椎软骨终板退变。进一步研究miR-140-5P不仅有利于揭示软骨终板退变的分子生物学机制,而且其日后也可能成为治疗椎间盘退变的有效靶工具。