Brain-Specific Cytoskeletal Damage Markers in Cerebrospinal Fluid: Is There a Common Pattern between Amyotrophic Lateral Sclerosis and Primary Progressive Multiple Sclerosis?

Many neurodegenerative disorders share a common pathophysiological pathway involving axonal degeneration despite different etiological triggers. Analysis of cytoskeletal markers such as neurofilaments, protein tau and tubulin in cerebrospinal fluid (CSF) may be a useful approach to detect the process of axonal damage and its severity during disease course. In this article, we review the published literature regarding brain-specific CSF markers for cytoskeletal damage in primary progressive multiple sclerosis and amyotrophic lateral sclerosis in order to evaluate their utility as a biomarker for disease progression in conjunction with imaging and histological markers which might also be useful in other neurodegenerative diseases associated with affection of the upper motor neurons. A long-term benefit of such an approach could be facilitating early diagnostic and prognostic tools and assessment of treatment efficacy of disease modifying drugs.     

Multistep Model of Cervical Cancer: Participation of miRNAs and Coding Genes

Aberrant miRNA expression is well recognized as an important step in the development of cancer. Close to 70 microRNAs (miRNAs) have been implicated in cervical cancer up to now, nevertheless it is unknown if aberrant miRNA expression causes the onset of cervical cancer. One of the best ways to address this issue is through a multistep model of carcinogenesis. In the progression of cervical cancer there are three well-established steps to reach cancer that we used in the model proposed here. The first step of the model comprises the gene changes that occur in normal cells to be transformed into immortal cells (CIN 1), the second comprises immortal cell changes to tumorigenic cells (CIN 2), the third step includes cell changes to increase tumorigenic capacity (CIN 3), and the final step covers tumorigenic changes to carcinogenic cells. Altered miRNAs and their target genes are located in each one of the four steps of the multistep model of carcinogenesis. miRNA expression has shown discrepancies in different works; therefore, in this model we include miRNAs recording similar results in at least two studies. The present model is a useful insight into studying potential prognostic, diagnostic, and therapeutic miRNAs.     

人巨细胞病毒pp65基因片段的原核表达及鉴定

目的构建人巨细胞病毒(HCMV)pp65基因片段(363～505位氨基酸)的原核表达质粒,并鉴定所表达蛋白的反应原性。方法以含有HCMVpp65全长基因的pGEM-T-pp65质粒为模板,PCR扩增pp65基因第1087～1515位核苷酸片段,酶切后插入pET-21a(+)载体,转化大肠杆菌BL21(DE3)。经IPTG诱导表达,纯化后进行Western blot鉴定。结果酶切分析和测序证明原核表达质粒pET-21a(+)-pp65构建正确。表达的融合蛋白主要以包涵体形式存在,以1.0mmol/L IPTG诱导5h,目的蛋白的表达量最高。纯化后的蛋白具有良好的反应原性。结论已成功构建了HCMV pp65基因片段原核表达质粒,并在大肠杆菌中表达了融合蛋白。     

Circulating Tumor Cells (CTC) and Cell-Free DNA (cfDNA) Workshop 2016: Scientific Opportunities and Logistics for Cancer Clinical Trial Incorporation

Despite the identification of circulating tumor cells (CTCs) and cell-free DNA (cfDNA) as potential blood-based biomarkers capable of providing prognostic and predictive information in cancer, they have not been incorporated into routine clinical practice. This resistance is due in part to technological limitations hampering CTC and cfDNA analysis, as well as a limited understanding of precisely how to interpret emergent biomarkers across various disease stages and tumor types. In recognition of these challenges, a group of researchers and clinicians focused on blood-based biomarker development met at the Canadian Cancer Trials Group (CCTG) Spring Meeting in Toronto, Canada on 29 April 2016 for a workshop discussing novel CTC/cfDNA technologies, interpretation of data obtained from CTCs versus cfDNA, challenges regarding disease evolution and heterogeneity, and logistical considerations for incorporation of CTCs/cfDNA into clinical trials, and ultimately into routine clinical use. The objectives of this workshop included discussion of the current barriers to clinical implementation and recent progress made in the field, as well as fueling meaningful collaborations and partnerships between researchers and clinicians. We anticipate that the considerations highlighted at this workshop will lead to advances in both basic and translational research and will ultimately impact patient management strategies and patient outcomes.     

Editorial on the Special Issue: Regulation by Non-Coding RNAs

This Special Issue of IJMS is devoted to regulation by non-coding RNAs and contains both original research and review articles. An attempt is made to provide an up-to-date analysis of this very fast moving field and cover regulatory roles of both microRNAs and long non-coding RNAs. Multifaceted functions of these RNAs in normal cellular processes, as well as in disease progression, are highlighted.     

原发性开角型青光眼致病基因MYOC真核表达质粒的构建及其在COS-7细胞中的表达

目的构建原发性开角型青光眼(POAG)致病基因MYOC的真核表达质粒,并在COS-7细胞中表达MYOC蛋白。方法用RT-PCR法扩增人眼组织(角膜缘)MYOC基因cDNA,纯化回收后,克隆入pGEM-T载体,再亚克隆入真核表达质粒pEGFP-N3,构建重组表达质粒pEGFP-N3-MYOC,转染COS-7细胞,用荧光显微镜观察MYOC蛋白在COS-7细胞中的表达,Western blot分析MYOC蛋白分泌特点。结果经酶切和DNA测序鉴定,证实重组表达质粒pEGFP-N3-MYOC构建正确,荧光显微镜观察MYOC蛋白能在COS-7细胞中表达,并且定位在细胞质中,而绿色荧光蛋白分布在整个细胞内。Western blot结果显示,MYOC蛋白能分泌到细胞外。结论已成功构建MYOC基因真核表达质粒,并能在COS-7细胞中表达MYOC蛋白,为进一步研究POAG发病机制奠定了基础。     

PPA多元醇相对分子质量测定的影响因素

概述了聚己二酸-1,2-丙二醇酯多元醇的几种相对分子质量的估算方法,讨论了这几种估算方法产生误差的原因和消除误差的对策。