超滤方法在体外循环中应用的探讨

目的：观察和研究超滤方法在体外循环心内直视手术中应用的效果。方法：统计自1989年9月至2002年1月间共实行8622例体外循环心内直视手术中应用超滤方法,其中男性4825例,女性3837例。体外循环中超滤法是在心内操作基本完成,且在机体给予复温时进行的。结果：全组病例超滤时间10-46分,超滤液量为300-1800ml,监测超滤液中未发现细胞成分,蛋白定性为阴性。超滤后血液得到浓缩,检测血液尿素氮,肌酐及炎性介质均比以前降低。结论：体外循环中应用超滤方法可以减轻病人术后组织水肿,并能迅速地浓缩血液,提高病人的血细胞压积,血红蛋白和各种凝血物质,超滤方法还能降低体外循环中的炎性介质的浓度,有利于患者术后的康复。     

Identification of the major TG4 cross‐linking sites in the androgen‐dependent SVS I exclusively expressed in mouse seminal vesicle

SVS I was exclusively expressed in seminal vesicle in which the protein was immunolocalized primarily to the luminal epithelium of mucosal folds. The developmental profile of its mRNA expression was shown to be androgen‐dependent, manifesting a positive correlation with the animal's maturation. There are 43 glutamine and 43 lysine residues in one molecule of SVS I, which is one of the seven major monomer proteins tentatively assigned on reducing SDS–PAGE during the resolution of mouse seminal vesicle secretion. Based on the fact that SVS I‐deduced protein sequence consists of 796 amino acid residues, we produced 7 recombinant polypeptide fragments including residues 1–78/F1, residues 79–259/F2, residues 260–405/F3, residues 406–500/F4, residues 501–650/F5, residues 651–715/F6, and residues 716–796/F7, and measured the covalent incorporation of 5‐(biotinamido)pentylamine (BPNH₂) or biotin‐TVQQEL (A25 peptide) to each of F1‐to‐F7 by type 4 transglutaminase (TG₄) from the coagulating gland secretion. F2 was active to a greater extent than the other fragments during the BPNH₂‐glutamine incorporation, and a relatively low extent of A25‐lysine cross link was observed with all of the seven fragments. The MS analysis of BPNH₂‐F2 conjugate identified Q²³² and Q²⁵⁴ as the two major TG₄ cross‐linking sites. This was substantiated by the result that much less BPNH₂ was cross‐linked to any one of the three F2 mutants, including Q232G and Q254G obtained from single‐site mutation, and Q232G/Q254G from double‐site mutation. J. Cell. Biochem. 107: 899–907, 2009. © 2009 Wiley‐Liss, Inc.     

肺炎克雷伯菌菌毛粘附素克隆表达及其对体外培养细胞的粘附活性

摘要：【目的】肺炎克雷伯菌(K.pn)与宿主细胞的粘附是致病的首要条件,粘附过程主要通过菌毛粘附素MrkD蛋白介导。为了进一步分析MrkD蛋白与宿主细胞间的粘附机制,进一步确定MrkD蛋白的粘附阻断作用。【方法】构建肺炎克雷伯菌菌毛粘附素融合蛋白原核表达质粒pGEX-4T-mrkD,转入大肠杆菌BL21,优化诱导表达条件,表达产物经亲和层析纯化、凝血酶切除融合蛋白GST标签后,进行SDS-PAGE和Western blot鉴定。激光共聚焦显微镜定位MrkD蛋白在宿主细胞上的结合部位;通过粘附活性试验与粘附动力学实验研究了MrkD蛋白的生物活性。【结果】实验得到了分子量为35 kDa的MrkD蛋白,定位了MrkD蛋白在宿主细胞上的结合部位,并证明了MrkD蛋白可以显著影响肺炎克雷伯菌对宿主细胞的粘附力。【结论】本试验首次证实了MrkD蛋白的粘附阻断作用并观察到了其与宿主细胞的作用位点,为研究肺炎克雷伯菌的致病机制,寻找粘附素功能表位奠定了基础。     

Cliques in mitotic spindle network bring kinetochore‐associated complexes to form dependence pathway

The mitotic spindle is an essential molecular machine for chromosome segregation during mitosis. Achieving a better understanding of its organization at the topological level remains a daunting task. To determine the functional connections among 137 mitotic spindle proteins, a protein–protein interaction network among queries was constructed. Many hub proteins, which connect more than one query and serve as highly plausible candidates for expanding the mitotic spindle proteome, are ranked by conventional degree centrality and a new subnetwork specificity score. Evaluation of the ranking results by literature reviews and empirical verification of SEPT6, a novel top‐ranked hub, suggests that the subnetwork specificity score could enrich for putative spindle‐related proteins. Topological analysis of this expanded network shows the presence of 30 3‐cliques and six 4‐cliques (fully connected subgraphs) that, respectively, reside in eight kinetochore‐associated complexes, of which seven are evolution conserved. Notably, these complexes strikingly form dependence pathways for the assembly of the kinetochore complex. These analyses indicate the feasibility of using network topology, i.e. cliques, to uncover novel pathways to accelerate our understanding of potential biological processes.     

Seasonal change and habitat selection of shorebird community at the South Yangtze River Mouth and North Hangzhou Bay, China

Coastal regions are important habitats for migratory shorebirds. The aim of the study is to understand habitat use by migratory shorebirds and to develop a conservation strategy in the sustainable use of wetlands. From March 2004 to January 2005, we conducted a seasonal shorebirds census in ten coastal habitats along the South Yangtze River mouth and North Hangzhou Bay, simultaneously examining the relative seasonal abundance of shorebirds and their spatial distribution. A total of 25 species were identified, the dominant seasonal species were Great Knot (Calidris tenuirostris), Sharp-tailed Sandpiper (Calidris alpine) and Red-necked Stint (Calidris ruficollis) in spring; Kentish Plover (Charadrius alexandrinus), Common Greenshank(Tringa nebularia) and Lesser Sand Plover (Charadrius mongolus) in summer; Kentish Plover, Red-necked Stint and Common Greenshank in autumn; Dunlin(Calidris alpine), Kentish Plover and Marsh Sandpiper (Tringa stagnatilis) in winter. These species accounted for more than 85% of the total shorebirds. The numbers of shorebirds counted was highest in spring and then in autumn, winter and summer respectively. Among the four seasons, there were few significant differences in the number of bird species between the sites outside the seawall (intertidal mudflat) and the sites inside the seawall (artificial wetland), but the average density of shorebirds was obviously different. The habitat-selection analysis of the environmental factors (outside and inside the seawall) impacting on the shorebird community was made in the 10 study sites with Canonical Correspondence Analysis. The study results indicated that: (1) Outside the seawall, the widths of the total intertidal mudflat and bare mudflat were the key factors affecting the shorebirds; the proportion of bulrush (Scirpus×mariquete) covering and supertidal mudflat width had a positive correlation with the abundance of birds, while human disturbance and the proportion of both reed (Phragmites communis) and smooth cord-grass (Spartina alterniflora) covering in total surveyed areas had negative impacts on bird numbers; (2) Inside the seawall, the proportions of areas with shallow water and mudflats occupying the total surveyed area were key factors influencing the number of birds; the size of the bulrush area should have a positive impact on the appearance of shorebirds. Habitats with heavy human disturbance, dense reed and smooth cord-grass or a high water level were not conducive to be inhabited by shorebirds.     

Response of vegetation phenology to urbanization in the conterminous United States

The influence of urbanization on vegetation phenology is gaining considerable attention due to its implications for human health, cycling of carbon and other nutrients in Earth system. In this study, we examined the relationship between change in vegetation phenology and urban size, an indicator of urbanization, for the conterminous United States. We studied more than 4500 urban clusters of varying size to determine the impact of urbanization on plant phenology, with the aids of remotely sensed observations since 2003–2012. We found that phenology cycle (changes in vegetation greenness) in urban areas starts earlier (start of season, SOS) and ends later (end of season, EOS), resulting in a longer growing season length (GSL), when compared to the respective surrounding urban areas. The average difference of GSL between urban and rural areas over all vegetation types, considered in this study, is about 9 days. Also, the extended GSL in urban area is consistent among different climate zones in the United States, whereas their magnitudes are varying across regions. We found that a tenfold increase in urban size could result in an earlier SOS of about 1.3 days and a later EOS of around 2.4 days. As a result, the GSL could be extended by approximately 3.6 days with a range of 1.6–6.5 days for 25th ~ 75th quantiles, with a median value of about 2.1 days. For different vegetation types, the phenology response to urbanization, as defined by GSL, ranges from 1 to 4 days. The quantitative relationship between phenology and urbanization is of great use for developing improved models of vegetation phenology dynamics under future urbanization, and for developing change indicators to assess the impacts of urbanization on vegetation phenology.     

An experimentally validated genome-scale metabolic reconstruction of Klebsiella pneumoniae MGH 78578, iYL1228

Klebsiella pneumoniae is a Gram-negative bacterium of the family Enterobacteriaceae that possesses diverse metabolic capabilities: many strains are leading causes of hospital-acquired infections that are often refractory to multiple antibiotics, yet other strains are metabolically engineered and used for production of commercially valuable chemicals. To study its metabolism, we constructed a genome-scale metabolic model (iYL1228) for strain MGH 78578, experimentally determined its biomass composition, experimentally determined its ability to grow on a broad range of carbon, nitrogen, phosphorus and sulfur sources, and assessed the ability of the model to accurately simulate growth versus no growth on these substrates. The model contains 1,228 genes encoding 1,188 enzymes that catalyze 1,970 reactions and accurately simulates growth on 84% of the substrates tested. Furthermore, quantitative comparison of growth rates between the model and experimental data for nine of the substrates also showed good agreement. The genome-scale metabolic reconstruction for K. pneumoniae presented here thus provides an experimentally validated in silico platform for further studies of this important industrial and biomedical organism.     

Distribution of the number of false discoveries in large-scale family-based association testing with application to the association between <Emphasis Type="Italic">PTPN1</Emphasis> and hypertension and obesity

We present a model-free approach to the study of the number of false discoveries for large-scale simultaneous family-based association tests (FBATs) in which the set of discoveries is decided by applying a threshold to the test statistics. When the association between a set of markers in a candidate gene and a group of phenotypes is studied by a class of FBATs, we indicate that a joint null hypothesis distribution for these statistics can be obtained by the fundamental statistical method of conditioning on sufficient statistics for the null hypothesis. Based on the joint null distribution of these statistics, we can obtain the distribution of the number of false discoveries for the set of discoveries defined by a threshold; the size of this set is referred to as its tail count. Simulation studies are presented to demonstrate that the conditional, not the unconditional, distribution of the tail count is appropriate for the study of false discoveries. The usefulness of this approach is illustrated by re-examining the association between PTPN1 and a group of blood-pressure-related phenotypes reported by Olivier et al. (Hum Mol Genet 13:1885–1892, 2004); our results refine and reinforce this association.