Differential ConA-enriched urinary proteome in rat experimental glomerular diseases

Glomerular diseases are leading causes of end-stage renal diseases worldwide. They are considered to be consequences of injury primarily to the three types of glomerular cells. Differential diagnosis typically relies on invasive biopsy findings. We expected that injuries of different glomerular cells would cause different changes in urinary proteome. The goal of this study was to identify differential urinary proteins distinguishing between injuries of different glomerular cells before significant histopathologic changes. Adriamycin nephropathy and Thy1.1 glomerulonephritis were employed as models with different primary impaired cells. ConA-enriched urinary glycoproteome on day3 were profiled by gel-free shotgun tandem mass spectrometry, and compared with self-healthy controls to identify differential urinary proteins for each model. By comparing the changes of the differential proteins between these two models, we identified 39 proteins with different directions of changes, which may potentially be useful in differentiation; and 7 proteins with the same direction of changes, which may be potential indicators of early renal damage. These differential proteins were of several origins: plasma proteins, proteins with urine or kidney specificity, proteins without tissue-specificity (mainly inflammatory mediators) etc. Our results may help better understand the effects of injuries of different glomerular cells at the initial stage, and lead to the discovery of novel early diagnostic markers for human focal segmental glomerulosclerosis (FSGS) and mesangioproliferative glomerulonephritis (MsPGN) which have the same primary impaired cells with adriamycin nephropathy and Thy1.1 glomerulonephritis, respectively.     

积水和网孔大小对青藏高原高寒沼泽化草甸优势种黑褐苔草叶片凋落物早期分解的影响

2013年5月,在青藏高原高寒沼泽化草甸收集优势种黑褐苔草(Carex atrofusca)立枯体凋落物。6月初,选择3个样带:长期积水带、周期性积水带和无积水带,并结合凋落物分解袋的方法,在每个样带区设置网孔大小分别为大(4.5 mm)、中(2.0 mm)、小(0.1mm)的3种凋落物分解袋,研究积水状态和网孔大小对凋落物早期分解的影响。结果显示:长期积水显著抑制凋落物的分解,而周期性积水和无积水环境中凋落物的质量损失率在大、中、小网孔凋落物袋中没有显著性的差异;网孔显著影响凋落物质量损失率,表现为大网孔中网孔小网孔;在早期分解过程中,凋落物的氮、磷均表现为富集;积水状态显著影响凋落物分解后氮、磷的富集;网孔大小对凋落物氮富集的影响显著,而对磷富集的影响不显著;积水状态和网孔大小对凋落物氮富集影响的交互作用极显著,但对磷富集没有显著的交互作用。研究发现,积水状态和网孔大小影响高寒沼泽化草甸凋落物分解以及氮、磷养分动态,由于不同网孔对凋落物分解的影响可用来反映不同土壤生物对凋落物分解的影响,因此,积水和土壤生物对维持高寒沼泽化草甸生态系统的结构和功能起着非常重要的作用。     

谷氧还蛋白系统及其对细胞氧化还原态势的调控

细胞内氧化还原调控主要是由谷氧还蛋白系统和硫氧还蛋白系统完成。谷氧还蛋白属于硫氧还蛋白超家族,广泛分布在各种生物体内。作为一种巯基转移酶,它能够催化巯基．二硫键交换反应或者还原蛋白质谷胱甘肽二硫化物,以维持胞内的氧化还原态势。谷氧蛋白系统参与氧化胁迫、蛋白修饰、信号转导、细胞调亡和细胞分化等多种生物过程。对其体内作用靶蛋白的研究,有助于阐明谷氧还蛋白在整个细胞氧化还原网络的重要调控作用。     

不同渍水时间对苗期和花期大豆生长及碳氮代谢的影响

以大豆品种南农99.6为材料,通过盆栽试验研究了苗期和花期渍水对大豆生长及碳氮代谢的影响.结果表明: 渍水显著抑制了大豆的生长,植株生物量、叶面积、叶片色素含量和光合速率均显著下降,而丙二醛(MDA)含量显著升高;随着渍水时间的延长,各生理指标的变化幅度增大;渍水胁迫解除后有一定的恢复,渍水10 d处理后恢复能力较渍水20 d处理强.可溶性碳、氮物质及关键酶对渍水反应不同,可溶性糖含量以及叶片谷氨酰胺合成酶和蔗糖合成酶的活性上升,而可溶性蛋白含量下降.渍水对苗期大豆植株生物量、叶面积和MDA含量的影响比花期小.苗期和花期渍水时间越短,大豆受到的伤害越小,其恢复能力也越强.渍水时间在10 d内,大豆植株能够通过自身的调节逐渐恢复.     

Expression and secretion of human bone morphogenetic protein-7 in pichia pastoris

The synonymous codons are used in a highly nonrandom manner in hosts of widely divergent species, which is termed ‘codon usage bias’. Several reports suggest that codon usage bias sometimes obstructs attempts to express high levels of exogenous genes. In this study, an attempt was made to express mature peptide of human bone morphogenetic protein-7 with optimized codons in P. pastoris expression system. Three low-usage ARG codons (CGG or CGA) in hBmp7 mature domain have been successfully transformed into P. pastoris-preferred ARG codons (AGA) with overlap extension PCR-based multiple-site-directed mutagenesis for a high level expression of hBMP7 mature peptide. The results of this study showed that the production level (25.45 mg/L) of a codon-optimized strain increased 4.6-fold in comparison with that (5.5 mg/L) of noncodon-optimized strain. A strain harboring multicopy of codon-optimized hbmp7 expression cassette showed an even higher expression level, which was about 2-fold compared with that of the single-copy one. These recombinant hBMP7 mature peptides were produced as 18-kD monomer proteins and were easily purified from culture supernatants using ion-exchange chromatography. Functional assay demonstrated that rhBMP7 could induce ectopic cartilage formation, although its inductive ability was much less active than that of CHO cell-derived hBMP7.     

定位于5q31-5q32的DFNA52的20个候选基因的突变筛查

为了克隆定位于5号染色体微卫星标记D5S2056和D5S638之间约8.8 cM的区间内的非综合征性常染色体显性遗传性耳聋 DFNA52 (OMIM: 607683)的致病基因, 文章根据基因在耳蜗组织的表达情况, 筛选出20个候选基因, 设计合成了扩增20个基因外显子及外显子与内含子交界的引物, 用DNA直接测序法进行序列变异分析。结果显示, 在基因外显子及侧翼区共发现了45个单核苷酸多态, 其中42个变异在多态数据库已报道, 其余3个为新发现的单核苷酸多态, 序列变异与疾病表型无共分离现象, 排除了这些基因外显子突变导致遗传性耳聋的可能性。     

我国重点氟污染行业排放清单与土壤氟浓度估算

工业生产排放和土壤氟高背景值导致我国部分地区氟污染严重,给生态安全和人类健康造成严重威胁.本文基于我国氟排放重点行业的生产产能,估算重点行业生产过程中氟排放量,构建我国重点行业氟排放清单,基于数据集成和融合,分析我国土壤氟背景值、氟地球化学分布和土壤氟浓度分布,并对氟污染典型区域氟污染成因及控制进行系统分析.分析发现,我国氟排放的重点行业有钢铁、磷肥和电解铝.磷肥施用的氟排放量最大,但施用面积大,浓度贡献小;电解铝行业的氟排放强度大;钢铁行业的氟排放总量大,但排放强度较小.我国大部分地区土壤氟背景值不高,环境容量大.但部分地区氟污染严重,主要是由工业氟排放和土壤氟高背景值造成,这些地区应采取相应的防控措施.     

渗透压对硝化作用的影响

在气升式内循环硝化反应器中研究了渗透压对硝化作用的影响。保持进水氨氮浓度420mg·L-1,将进水渗透压逐渐从4.3×105Pa提高到18.8×105Pa,硝化反应器的氨氮转化率稳定在93%～100%。将进水渗透压进一步提高到19.2×105Pa,氨氮转化率降至69.2%。渗透压对硝化作用的影响具有突发性,临界值在18.8×105～19.2×105Pa之间。受高渗透压胁迫时,活性污泥中硝化细菌的形态趋向单一,个体变小,内膜数量减少,并产生许多不明成分的颗粒状内含物。解除渗透压胁迫后,细胞结构恢复。添加钾离子能够缓解高渗压对硝化作用的影响。高渗透压胁迫以及解除渗透压胁迫可增强污泥硝化活性,比污泥氨氮转化率(污泥以SS计)分别从0.083kg·kg-1·d-1升至0.509kg·kg-1·d-1和2.569kg·kg-1·d-1,同比提高5.1倍和30.0倍。     

干酪乳杆菌arat基因的敲除及其对苯乳酸合成代谢的影响

苯乳酸（PLA）作为一种新型的广谱抑菌物质,在食品行业具有巨大的发展潜力,作为乳酸菌的天然代谢产物之一,对其代谢机理的研究具有非常重要的意义。通过CRISPR/Cas9基因编辑系统对一株干酪乳杆菌（Lactobacillus casei）1.8727的芳香族氨基转移酶基因arat进行敲除,得到一株arat缺失菌株（L. casei）1.8727Δarat。通过HPLC方法分析其代谢产物,发现发酵72 h时,PLA产量比出发菌株提高约66.7%,而苯丙氨酸（Phe）比出发菌株提高约57.8%。首次在L. casei 1.8727中成功应用基因编辑手段敲除了arat基因,研究表明该菌株具有自身合成代谢Phe的能力,arat作为PLA代谢途径的关键酶基因,其缺失并未使PLA产量下调,而是促进了PLA与Phe的合成代谢,证明干酪乳杆菌中可能存在其他的代偿途径,arat基因缺失所造成的代谢流的改变最终造成PLA与Phe产量的提高,同时PLA的合成代谢可能涉及多个基因的参与,是一个复杂的代谢网络。研究结果对于深入研究PLA的合成代谢机制具有重要意义,为后续PLA合成代谢途径提供了新的思路及方法。