基于免疫纳米颗粒强化的压电免疫传感器检测大肠杆菌O157︰H7

摘要：【目的】结合纳米技术建立检测大肠杆菌（Escherichia coli）O157︰H7高灵敏检测技术。【方法】采用化学共沉淀法制备出核心粒径约为10 nm的免疫纳米磁颗粒,柠檬酸钠还原法制备粒径约为20 nm的免疫胶体金。压电免疫传感器通过金黄色葡萄球菌蛋白A（Protein A from Staphylococcus aureus SPA）法将抗体固定于石英晶振上,两种免疫纳米颗粒借助不同的抗体连接于传感器上对检测频率信号进行放大。【结果】SPA在石英晶振上的最佳固定浓度和时间为1.2 mg/mL和40 min,抗体的最佳固定浓度和时间为1.0 mg/mL和60 min。压电免疫传感器通过两种免疫纳米颗粒的放大作用,使其对大肠杆菌O157︰H7的检测限从104 cfu/mL提高到101 cfu/mL。【结论】免疫纳米颗粒强化对压电免疫传感器的检测频率信号具有很好的放大效应,可以明显提高其检测灵敏度。     

Promoting human embryonic stem cell renewal or differentiation by modulating Wnt signal and culture conditions

We previously showed that Wnt3a could stimulate human embryonic stem （hES） cell proliferation and affect cell fate determination. In the absence of feeder cell--derived factors, hES cells cultured under a feeder-free condition survived and proliferated poorly. Adding recombinant Wnt3a in the absence of feeder cell derived-factors stimulated hES cell proliferation but also differentiation. In the present study, we further extended our analysis to other Wnt ligands such as Wntl and Wnt5a. While Wntl displayed a similar effect on hES cells as Wnt3a, Wnt5a had little effect in this system. Wnt3a and Wntl enhanced proliferation of undifferentiated hES cells when feeder-derived self-renewal factors and bFGF are also present. To explore the possibility to promote the proliferation of undifferentiated hES cells by activating the Wnt signaling, we overexpressed Wnt3a or Wntl gene in immortalized human adult fibroblast （HAFi） cells that are superior in supporting long-term growth of undifferentiated hES cells than primary mouse embryonic fibroblasts. HAFi cells with or without a Wnt tmnsgene can be propagated indefinitely. Over-expression of the Wnt3a gene significantly enhanced the ability of HAFi feeder cells to support the undifferentiated growth of 3 different hES cell lines we tested. Co-expression of three commonly-used drug selection genes in Wnt3a-overpressing HAFi cells further enabled us to select rare hES clones after stable transfection or transduction. These immortalized engineered feeder cells （W3R） that co-express growth-promoting genes such as Wnt3a and three drug selection genes should empower us to efficiently make genetic modified hES cell lines for basic and translational research.     

基于RS的湖岸带物理结构完整性评价方法——以查干湖为例

以查干湖流域为研究区,利用RS和GIS技术,基于水利部《河流(湖)健康评估指标、标准与方法 V1.0》并加以改进,以500m×1000m为评价基本单元,整个研究区共有315个物理结构基本评价单元,建立基于RS的湖岸带物理结构完整性评价体系,该评价体系由目标层、准则层和指标层构成,其中准则层由湖岸带状况、湖岸线发育率和湖泊萎缩率3项组成,指标层由地形坡度、植被覆盖率等8项指标构成。研究结果表明:38个监测点中,查干湖23个,新庙泡15个,基于RS物理结构评价结果分别为0.67—0.76和0.35—0.45,地面实测评价结果分别是0.64—0.77和0.35—0.55。两种评价方法结果一致,并表明查干湖的湖岸带物理结构健康状态属于健康,新庙泡则属于亚健康。     

大鼠脂肪间充质干细胞诱导为类肝细胞及其细胞片的制备

成体多能干细胞,如来自骨髓和脂肪组织的间充质干细胞等具有多向分化的潜能。虽然自体干细胞移植已经发展成为器官移植的有效代替疗法之一,但是由于移植位点细胞的流失和分化条件的限制等问题使得这种疗法的效率大大降低。本研究目的是将由脂肪干细胞分化而来的类肝细胞制备成具有稳定细胞性状的可移植的肝细胞片。首先在体外分离扩增脂肪干细胞,并通过控制严格地分化条件获得类肝细胞。然后将此细胞接种到聚N-异丙基丙烯酰胺(PNIPAAm)结合的细胞培养皿表面,通过调节培养温度到20oC,使细胞成片脱离培养皿形成细胞片。对细胞片进行了常规HE染色和免疫组化观察,结果显示:这类细胞片中平均含有2～3层细胞,并且保持了细胞外基质的完整。同传统的胰酶消化收集移植用细胞相比,细胞片方法极大地减少了对移植用细胞的细胞膜和细胞外基质的损伤,这将大大促进细胞片和原位组织的相互作用,增加细胞利用效率,从而有望提高治疗效果。     

Lysophosphatidic Acid Acyltransferase from Coconut Endosperm Mediates the Insertion of Laurate at the sn-2 Position of Triacylglycerols in Lauric Rapeseed Oil and Can Increase Total Laurate Levels

Expression of a California bay laurel (Umbellularia californica) 12:0-acyl-carrier protein thioesterase, bay thioesterase (BTE), in developing seeds of oilseed rape (Brassica napus) led to the production of oils containing up to 50% laurate. In these BTE oils, laurate is found almost exclusively at the sn-1 and sn-3 positions of the triacylglycerols (T.A. Voelker, T.R. Hayes, A.C. Cranmer, H.M. Davies [1996] Plant J 9: 229–241). Coexpression of a coconut (Cocos nucifera) 12:0-coenzyme A-preferring lysophosphatitic acid acyltransferase (D.S. Knutzon, K.D. Lardizabal, J.S. Nelsen, J.L. Bleibaum, H.M. Davies, J.G. Metz [1995] Plant Physiol 109: 999–1006) in BTE oilseed rape seeds facilitates efficient laurate deposition at the sn-2 position, resulting in the acccumulation of trilaurin. The introduction of the coconut protein into BTE oilseed rape lines with laurate above 50 mol % further increases total laurate levels.     

Relationships between biological soil crusts,bacterial diversity and abundance,and ecosystem functioning: Insights from a semi‐arid Mediterranean environment

Questions: To what degree do biological soil crusts (BSCs), which are regulators of the soil surface boundary, influence associated microbial communities? Are these associations important to ecosystem functioning in a Mediterranean semi‐arid environment? Location: Gypsum outcrops near Belmonte del Tajo, Central Spain. Methods: We sampled a total of 45 (50 cm × 50 cm) plots, where we estimated the cover of every lichen and BSC‐forming lichen species. We also collected soil samples to estimate bacterial species richness and abundance, and to assess different surrogates of ecosystem functioning. We used path analysis to evaluate the relationships between the richness/abundance of above‐ and below‐ground species and ecosystem functioning. Results: We found that the greatest direct effect upon the ecosystem function matrix was that of the biological soil crust (BSC) richness matrix. A few bacterial species were sensitive to the lichen community, with a disproportionate effect of Collema crispum and Toninia sedifolia compared to their low abundance and frequency. The lichens Fulgensia subbracteata and Toninia spp. also had negative effects on bacteria, while Diploschistes diacapsis consistently affected sensitive bacteria, sometimes positively. Despite these results, very few of the BSC effects on ecosystem function could be ascribed to changes within the bacterial community. Conclusion: Our results suggest the primary importance of the richness of BSC‐forming lichens as drivers of small‐scale changes in ecosystem functioning. This study provides valuable insights on semi‐arid ecosystems where plant cover is spatially discontinuous and ecosystem function in plant interspaces is regulated largely by BSCs.     

聚乙烯醇生物降解研究进展

聚乙烯醇(PVA)是一种在纺织和化工行业中广泛使用的难降解的高分子聚合物。随着人们对纺织工业清洁生产的关注,如何在退浆工艺中就实现对PVA的生物降解、减少PVA废水的排放,并避免化学退浆过程中高温和氧化造成的棉纤维损伤,是近年来纺织生物技术领域的研究热点。由于PVA降解菌种类不多、培养周期长,PVA降解酶酶活不高、提取不容易等原因,使PVA的生化降解研究还局限在PVA降解菌的筛选、PVA降解酶的酶学性质研究等方面,PVA降解酶还未在纺织工业上得到应用。本文综述了近年来国内外在PVA降解菌筛选、PVA降解酶提取及酶学性质、PVA生化降解机理等方面的研究进展,并讨论了PVA生化降解研究中存在的问题及发展方向。     

Effects of size,neighbors, and site condition on tree growth in a subtropical evergreen and deciduous broad‐leaved mixed forest,China

Successful growth of a tree is the result of combined effects of biotic and abiotic factors. It is important to understand how biotic and abiotic factors affect changes in forest structure and dynamics under environmental fluctuations. In this study, we explored the effects of initial size [diameter at breast height (DBH)], neighborhood competition, and site condition on tree growth, based on a 3‐year monitoring of tree growth rate in a permanent plot (120 × 80 m) of montane Fagus engleriana–Cyclobalanopsis multiervis mixed forest on Mt. Shennongjia, China. We measured DBH increments every 6 months from October 2011 to October 2014 by field‐made dendrometers and calculated the mean annual growth rate over the 3 years for each individual tree. We also measured and calculated twelve soil properties and five topographic variables for 384 grids of 5 × 5 m. We defined two distance‐dependent neighborhood competition indices with and without considerations of phylogenetic relatedness between trees and tested for significant differences in growth rates among functional groups. On average, trees in this mixed montane forest grew 0.07 cm year^?1 in DBH. Deciduous, canopy, and early‐successional species grew faster than evergreen, small‐statured, and late‐successional species, respectively. Growth rates increased with initial DBH, but were not significantly related to neighborhood competition and site condition for overall trees. Phylogenetic relatedness between trees did not influence the neighborhood competition. Different factors were found to influence tree growth rates of different functional groups: Initial DBH was the dominant factor for all tree groups; neighborhood competition within 5 m radius decreased growth rates of evergreen trees; and site condition tended to be more related to growth rates of fast‐growing trees (deciduous, canopy, pioneer, and early‐successional species) than the slow‐growing trees (evergreen, understory, and late‐successional species).