Expression of Dystrophin in Duchenne Muscular Dystrophy Patients after Myoblast Transplantation

Based on originally designed technique of myoblast cultivation and in accordance with the approved by the Russian Ministry of Health one muscle treatment protocol of myoblast transplantation to the Duchenne muscular dystrophy patients, the first in Russia clinical trial of this gene correction method was carried out. Immunologically related myoblast cultures (30 to 90 million cells per patient) were injected after all preliminary procedures into tibialis anterior muscles of four boys selected from a group of volunteer recipients (Duchenne muscular dystrophy patients) based on the analysis of a number of surface antigens in donor–recipient pairs. The condition of the patients remained satisfactory during the whole period of post-transplantation follow-up (from 6 months to 1.5 years). Six months after myoblast transplantation the presence of donor DNA or dystrophin synthesis was demonstrated in muscle biopsies of three out of four patients. This result confirms efficacy and safety of the procedure used.     

Interactions between the Su(Hw) and Mod(mdg4) proteins required for gypsy insulator function

The gypsy insulator is thought to play a role in nuclear organization and the establishment of higher order chromatin domains by bringing together several individual insulator sites to form rosette-like structures in the interphase nucleus. The Su(Hw) and Mod(mdg4) proteins are components of the gypsy insulator required for its effect on enhancer-promoter interactions. Using the yeast two-hybrid system, we show that the Mod(mdg4) protein can form homodimers, which can then interact with Su(Hw). The BTB domain of Mod(mdg4) is involved in homodimerization, whereas the C-terminal region of the protein is involved in interactions with the leucine zipper and adjacent regions of the Su(Hw) protein. Analyses using immunolocalization on polytene chromosomes confirm the involvement of these domains in mediating the interactions between these proteins. Studies using diploid interphase cells further suggest the contribution of these domains to the formation of rosette-like structures in the nucleus. The results provide a biochemical basis for the aggregation of multiple insulator sites and support the role of the gypsy insulator in nuclear organization.     

Plant Resistance Suppressors in the Pathosystem Formed by Potato and the Causal Agent of Late Blight

The properties and effects of two plant resistance suppressors (1,3--1,6--glucan and a pentasaccharide of xyloglucan origin) involved in the pathosystem of potato (Solanum tuberosum) and the causal agent of blight (Phytophthora infestans(Mont) de Bary) were compared. The microbial 1,3--1,6--glucan suppressed the defense response over a narrow concentration range (10^–2M), whereas the plant pentasaccharide had a broad range of effective concentrations (10^–12to 10^–6M). In the pathosystem of potato and the causal agent of late blight, the -glucan caused a local and race-specific suppressor effect on the plant host defense response. In contrast, the pentasaccharide caused both local and systemic suppression of potato resistance and the presence of terminal fucosyl residue in the xyloglucan oligosaccharine played a decisive role in its effect. The recognition of both suppressors by potato cell membrane sites is discussed.     

硫氧还蛋白抗体柱的制备

目的:探索硫氧还蛋白(Trx)抗体柱对Trx融合蛋白纯化的可行性。方法与结果:对含有Trx基因的质粒表达载体pTrxFus进行改造,在Trx读框之后加入6×His序列,并在大肠杆菌中表达C端带有6×His标签的Trx,经Ni2+柱亲和纯化后制备多克隆抗体;把经蛋白A纯化后的抗体偶联在溴化氰活化的琼脂糖凝胶上,制成Trx抗体柱;用此抗体柱纯化与Trx融合表达的豇豆胰蛋白酶抑制剂(CpTI),SDS-PAGE结果显示获得了纯度较高的Trx-CpTI。结论:用Trx抗体制成的免疫亲和层析柱可以有效纯化Trx融合蛋白。     

创伤患者急性肾功能损伤诊断与治疗

目的:探讨患者创伤后发生急性肾损伤的患者发病率、临床特点以及发病危险因素,以便有效预防和及早治疗。方法:回顾性分析我院重症监护室2004年1月至2010年12月收治的创伤患者相关临床资料,分析创伤后急性肾损伤的发病率以及发病危险因素。结果:共有106例患者纳入我们的研究,其中47例患者创伤后并发急性肾损伤。在发生急性肾损伤患者中,平均年龄为31±19岁,84.6%为男性;其中25例为脓毒血症引起,18例是因为低血压导致急性肾功能损伤。所有患者中,24例患者出现了少尿的症状,19例患者进行了透析治疗。腹部外伤[(OR)=3.66,P=0.027]和应用呋塞米[(OR=4.10,P=0.026)]是发生急性肾损伤的危险因素。结论:急性肾损伤时创伤后的严重并发症之一,死亡率高。只有找到创伤后发生急性肾损伤的危险因素,才能有效预防和及早治疗。     

中药夏枯草对人甲状腺癌细胞系SW579增殖周期及凋亡的影响

目的:探讨夏枯草对人甲状腺癌细胞系SW579细胞生长的抑制作用及其对细胞增殖周期和凋亡的影响。方法:采用甲基噻唑(MTT)比色法和生长曲线测定不同浓度夏枯草在不同作用时间内对在体外培养SW579细胞增殖的影响,同时应用流式细胞术检测细胞增殖周期及凋亡率的变化。结果:夏枯草可在G0/G1期阻滞人甲状腺癌细胞系SW579的增殖,使S期细胞比率降低;在一定范围内,夏枯草的浓度越高、作用时间越长,对肿瘤细胞生长的抑制作用越强,凋亡率也越高。结论:夏枯草能抑制人甲状腺癌细胞系SW579细胞生长,并诱导细胞凋亡而阻止细胞周期。     

Kmasker plants – a tool for assessing complex sequence space in plant species

Many plant genomes display high levels of repetitive sequences. The assembly of these complex genomes using short high‐throughput sequence reads is still a challenging task. Underestimation or disregard of repeat complexity in these datasets can easily misguide downstream analysis. Detection of repetitive regions by k‐mer counting methods has proved to be reliable. Easy‐to‐use applications utilizing k‐mer counting are in high demand, especially in the domain of plants. We present Kmasker plants, a tool that uses k‐mer count information as an assistant throughout the analytical workflow of genome data that is provided as a command‐line and web‐based solution. Beside its core competence to screen and mask repetitive sequences, we have integrated features that enable comparative studies between different cultivars or closely related species and methods that estimate target specificity of guide RNAs for application of site‐directed mutagenesis using Cas9 endonuclease. In addition, we have set up a web service for Kmasker plants that maintains pre‐computed indices for 10 of the economically most important cultivated plants. Source code for Kmasker plants has been made publically available at https://github.com/tschmutzer/kmasker . The web service is accessible at https://kmasker.ipk-gatersleben.de .