Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites

The investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci – six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.     

基于生态敏感性评价的本溪青云山景区空间规划

林地型景区的生态环境保护和利用是景区旅游开发的基础.本文基于生态环境要素和保护需要构建了生态敏感性评价指标体系,通过层次分析法和熵权法得到组合权重,结合GIS技术对本溪青云山景区进行生态敏感性评价,并根据评价结果制定景区空间规划.结果表明:青云山景区内高敏感区占比9.2%,敏感区占比60.2%,低敏感区占比22.7%,不敏感区占比7.9%.依据青云山生态特征和生态保护红线将景区分为限制开发区、适度开发区和旅游开发区三大板块,结合自然、人文等资源的空间分布特征,制定了景区"二轴六区"的空间规划,针对各板块资源利用现状和存在的问题,制定了相应的旅游开发规划策略.本研究能够为林地型景区空间规划提供更为科学有效的规划路径.     

Genomic overview of serine proteases

Serine proteases (SP) are peptidases with a uniquely activated serine residue in the substrate-binding pocket. They represent about 0.6% of all proteins in the human genome. SP are involved in many vital functions such as digestion, blood clotting, fibrinolysis, fertilization, and complement activation and are related to many diseases including cancer, arthritis, and emphysema. In this study, we performed a genomic analysis of human serine proteases utilizing different databases, primarily that of MEROPS. SP are distributed along all human chromosomes except 18 and Y with the highest density (23 genes) on chromosome 19. They are either randomly located within the genome or occur in clusters. We identified a number of SP clusters, the largest being the kallikrein cluster on chromosome 19q13.4 which is formed of 15 adjacent genes. Other clusters are located on chromosomes 19p13, 16p13, 14q11, 13q35, 11q22, and 7q35. Genes of each cluster tend to be of comparable sizes and to be transcribed in the same direction. The members of some clusters are sometimes functionally related, e.g., the involvement of many kallikreins in endocrine-related malignancies and the hematopoietic cluster on chromosome 14. It is hypothesized that members of some clusters are under common regulatory mechanisms and might be involved in cascade enzymatic pathways. Several functional domains are found in SP, which reflect their functional diversity. Membrane-type SP tend to cluster in 3 chromosomes and have some common structural domains. Several databases are available for screening, structural and functional analysis of serine proteases. With the near completion of the Human Genome Project, research will be more focused on the interactions between SP and their involvement in pathophysiological processes.     

The Oompa-Loompas of the genome factories

Automation Technologies for Genome Characterization

edited by Tony J. Beugelsdijk, John Wiley and Sons, 1997. UK£55.00 hbk (xvi +306 pages) ISBN 0 471 12806 6     

Evidence of colorectal cancer risk associated variant Lys25Ser in the proximity of human bone morphogenetic protein 2

Colorectal cancer (CRC) is the third most prevalent cancer and fourth leading cause of cancer-related deaths globally. It has been shown that the nsSNP variants play an important role in diseases, however it remained unclear how these variants are associated with the disease. Recently, several CRC risk associated SNPs have been discovered, however rs961253 (Lys25Arg at 20p12.3) located in the proximity of bone morphogenetic protein 2 (Bmp2) and fermitin family homolog 1 Fermt1 genes have been reported to be highly associated with the CRC risk. Here we provide evidence for the first time in silico biological functional and structural implications of non-synonymous (nsSNPs) CRC disease-associated variant Lys25Arg via molecular dynamic (MD) simulation. Protein structural analysis was performed with a particular variant allele (A/C, Lys25Arg) and compared with the predicted native protein structure. Our results showed that this nsSNP will cause changes in the protein structure and as a result is associated with the disease. In addition to the native and mutant 3D structures of CRC associated risk allele protein domain (CRAPD), they were also analyzed using solvent accessibility models for further protein stability confirmation. Taken together, this study confirmed that this variant has functional effect and structural impact on the CRAPD and may play an important role in CRC disease progression; hence it could be a reasonable approach for studying the effect of other deleterious variants in future studies.     

我国医疗服务项目成本核算研究述评 ——基于演变历程的视角

目的 了解我国医疗服务项目成本核算文献研究进展和政策推进情况。方法 检索中国知网、万方数据库、重庆维普数据库1978—2016年医疗服务项目成本核算相关文献和政策文件,进行内容分析。结果 通过对文献研究主题、数量与时间分布的三维匹配,将我国医疗服务项目成本核算演变历程分为起步（1979—1991年）、探索前期（1992—2001年）、探索中期（2002—2009年）、探索后期（2010年至今）4个阶段。结论 不同时期医疗服务项目成本核算政策推进均滞后于学术研究,缺乏政策导向性;医疗服务项目成本核算仍处于探索阶段,亟待标准化;核算技术标准化是医疗服务项目成本核算标准化的核心;未来医疗服务项目成本核算功能应向医疗质量安全成本一体化扩展;夯实核算基础与完善配套政策是开展医疗服务项目成本核算的关键。     

国家自然科学基金委重大项目“禾本科植物的适应性辐射及其进化机制”结题综述

国家自然科学基金委重大项目"禾本科植物的适应性辐射及其进化机制"利用比较形态学、分子系统学、进化发育生物学、古生物学等多方面证据,通过多学科交叉的方法探讨了禾本科植物中出现的适应性辐射现象及其机制,在禾本科的系统发育关系以及适应性辐射的基本式样、生物和非生物环境因素在物种适应和分化以及物种快速形成中的作用、基因组大小及其结构变异以及突变、重复和调控模式变化对适应性辐射过程中关键性状(功能)的影响等方面取得了重要进展,在人才培养、国际合作和实验体系建立等方面极具特点。该项目的顺利完成为更好地阐明植物多样性形成的原因与机理奠定了良好的基础。     

鼠Ⅰ型可溶性白细胞介素1受体基因在昆虫细胞的克隆和表达

白细胞介素１（ＩＬ－１）是一种重要的细胞因子,具有广泛的生物学活性。它通过与细胞表面的白细胞介素 １受体（ＩＬ－１Ｒ）结合而起作用。以杆状病毒为载体在昆虫细胞中克隆表达了小鼠Ｉ型可溶性白细胞介素１受体（ｓＩＬ－１　ＲＩ）基因。以ＮＩＨ／３Ｔ３细胞ＲＮＡ为模板,采用ＲＴ－ＰＣＲ方法扩增得到小鼠ｓＩＬ－ＩＲＩ的ｃＤＮＡ,克隆至杆状病毒转移载体ｐＡｃＧＰ６７Ｂ,将转移重组质粒与野生病毒ＡＣＮＰＶ ＤＮＡ共转染昆虫细胞Ｓｆ９,经同源重组得到重组杆状病毒ｒＡＣＮＰＶ。应用经纯化的ｒＡｃＮＰＶ感染昆虫细胞Ｓｆ９,表达获得重组的ｓＩＬ－１ＲＩ。经对亲和层析样品的ＳＤＳ－ＰＡＧＥ分析和对ＩＬ－１β生物活性阻断作用实验证实,表达产物能够与其配基结合,并且能够分泌至细胞培养上清中。     

2000-2019年京津冀地区植被覆盖状况变化及驱动因素

近20年来中国和印度通过土地利用活动改变地表覆盖使植被变得更绿,京津冀地区是植被变得更绿的典型地区。收集京津冀地区2000-2019年MODIS增强型植被指数（EVI）、植被覆盖百分比数据,以及年平均温度和降水量等数据,分析该区近20年来自然植被和农田植被EVI变化过程和趋势,揭示其变化的驱动因素,结果显示2000-2019年京津冀地区自然植被和农田植被EVI显著增加,自然植被EVI增加速率是农田植被的1.8倍。99.51%的自然植被和96.95%的农田植被生长状况改善。2000-2019年京津冀地区自然植被EVI与年平均温度和年降水量相关性不显著。农田植被EVI与年平均温度显著相关,与年降水量相关性不显著。农田灌溉和年平均温度变化是农田植被EVI显著增加的主要驱动因素。近20年京津冀地区通过实施以造林为主的生态建设工程,自然植被生长覆盖状况呈现极显著变好。同时森林植被比非森林植被覆盖百分比增加趋势更明显。以造林为主的生态建设工程是京津冀地区自然植被生长覆盖状况显著变好的主要驱动因素。     

Projecting the distribution of forests in New England in response to climate change

Aim To project the distribution of three major forest types in the northeastern USA in response to expected climate change. Location The New England region of the United States. Methods We modelled the potential distribution of boreal conifer, northern deciduous hardwood and mixed oak–hickory forests using the process‐based BIOME4 vegetation model parameterized for regional forests under historic and projected future climate conditions. Projections of future climate were derived from three general circulation models forced by three global warming scenarios that span the range of likely anthropogenic greenhouse gas emissions. Results Annual temperature in New England is projected to increase by 2.2–3.3 °C by 2041–70 and by 3.0–5.2 °C by 2071–99 with corresponding increases in precipitation of 4.7–9.5% and 6.4–11.4%, respectively. We project that regional warming will result in the loss of 71–100% of boreal conifer forest in New England by the late 21st century. The range of mixed oak–hickory forests will shift northward by 1.0–2.1 latitudinal degrees (c. 100–200 km) and will increase in area by 149–431% by the end of the 21st century. Northern deciduous hardwoods are expected to decrease in area by 26% and move upslope by 76 m on average. The upslope movement of the northern deciduous hardwoods and the increase in oak–hickory forests coincide with an approximate 556 m upslope retreat of the boreal conifer forest by 2071–99. In our simulations, rising atmospheric CO₂ concentrations reduce the losses of boreal conifer forest in New England from expected losses based on climatic change alone. Main conclusion Projected climate warming in the 21st century is likely to cause the extensive loss of boreal conifer forests, reduce the extent of northern hardwood deciduous forests, and result in large increases of mixed oak–hickory forest in New England.