Novel Staphylococcal Glycosyltransferases SdgA and SdgB Mediate Immunogenicity and Protection of Virulence-Associated Cell Wall Proteins

Infection of host tissues by Staphylococcus aureus and S. epidermidis requires an unusual family of staphylococcal adhesive proteins that contain long stretches of serine-aspartate dipeptide-repeats (SDR). The prototype member of this family is clumping factor A (ClfA), a key virulence factor that mediates adhesion to host tissues by binding to extracellular matrix proteins such as fibrinogen. However, the biological siginificance of the SDR-domain and its implication for pathogenesis remain poorly understood. Here, we identified two novel bacterial glycosyltransferases, SdgA and SdgB, which modify all SDR-proteins in these two bacterial species. Genetic and biochemical data demonstrated that these two glycosyltransferases directly bind and covalently link N-acetylglucosamine (GlcNAc) moieties to the SDR-domain in a step-wise manner, with SdgB appending the sugar residues proximal to the target Ser-Asp repeats, followed by additional modification by SdgA. GlcNAc-modification of SDR-proteins by SdgB creates an immunodominant epitope for highly opsonic human antibodies, which represent up to 1% of total human IgG. Deletion of these glycosyltransferases renders SDR-proteins vulnerable to proteolysis by human neutrophil-derived cathepsin G. Thus, SdgA and SdgB glycosylate staphylococcal SDR-proteins, which protects them against host proteolytic activity, and yet generates major eptopes for the human anti-staphylococcal antibody response, which may represent an ongoing competition between host and pathogen.     

Functional analysis of a miRNA‐like small RNA derived from Bombyx mori cytoplasmic polyhedrosis virus

Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is a major pathogen of the economic insect silkworm, Bombyx mori. Virus‐encoded microRNAs (miRNAs) have been proven to play important roles in host–pathogen interactions. In this study we identified a BmCPV‐derived miRNA‐like 21 nt small RNA, BmCPV‐miR‐1, from the small RNA deep sequencing of BmCPV‐infected silkworm larvae by stem‐loop quantitative real‐time PCR (qPCR) and investigated its functions with qPCR and lentiviral expression systems. Bombyx mori inhibitor of apoptosis protein (BmIAP) gene was predicted by both target prediction software miRanda and Targetscan to be one of its target genes with a binding site for BmCPV‐miR‐1 at the 5′ untranslated region. It was found that the expression of BmCPV‐miR‐1 and its target gene BmIAP were both up‐regulated in BmCPV‐infected larvae. At the same time, it was confirmed that BmCPV‐miR‐1 could up‐regulate the expression of BmIAP gene in HEK293T cells with lentiviral expression systems and in BmN cells by transfecting mimics. Furthermore, BmCPV‐miR‐1 mimics could up‐regulate the expression level of BmIAP gene in midgut and fat body in the silkworm. In the midgut of BmCPV‐infected larvae, BmCPV‐miR‐1 mimics could be further up‐regulated and inhibitors could lower the virus‐mediated expression of BmIAP gene. With the viral genomic RNA segments S1 and S10 as indicators, BmCPV‐miR‐1 mimics could up‐regulate and inhibitors down‐regulate their replication in the infected silkworm. These results implied that BmCPV‐miR‐1 could inhibit cell apoptosis in the infected silkworm through up‐regulating BmIAP expression, providing the virus with a better cell circumstance for its replication.     

无血清培养高密度乳猪肝细胞的形态和功能变化

本文研究了无血清培养高密度猪肝细胞的形态和功能变化。将分离的肝细胞以高密度(1×10~7/ml)培养在含激素、多种生长因子和营养成分的无血清培养基中,动态观察培养7天中肝细胞形态、活率、蛋白质合成功能、G-6-Pase活性、安定转化功能及LDH含量;同时以无血清培养低密度(5×10~5/ml)肝细胞作为对照组。研究结果表明:高密度培养的 肝细胞各项功能较低密度培养的肝细胞为低;高密度培养的肝细胞的形态、蛋白质合成功能在培养7天中保持稳定;活率随着培养时间的延长而下降,但均高于90％;安定转化功能在培养第2、3天最强;G-6-Pase活性在培养1天后明显下降,然后维持在较低水平;LDH含量在第1、2、3天较高。     

香溪河水系大型底栖动物功能摄食类群生态学

2004年7月至2007年6月,通过大型底栖无脊椎动物的量化监测,对三峡水库湖北库区最大河流香溪河大型底栖无脊椎动物功能摄食类群生态学进行研究.结果表明:香溪河大型底栖动物以收集者占绝对优势,其次为刮食者,捕食者、滤食者、撕食者相对丰度较小;各功能摄食类群分布明显受时空资源位的限制;香溪河物质循环能力、两岸物质的输入量和粗有机颗粒/ 细有机颗粒在九冲河明显高于其它河流,在冬季高于其它季节;物质输送能力以高岚河最高,在时间方面以冬季明显高于其它季节.逐步回归分析表明,流速、电导、浊度、总氮、二氧化硅对香溪河河流生态系统物质循环具有指示作用;二氧化硅和化学需氧量对物质输送能力具有指示作用;水温、水深、二氧化硅、总磷可作为沿岸物质输入量的指标;水温、二氧化硅可作为评估河道中粗、细有机颗粒比例的重要指标.     

Recent advances in CRISPR/Cas9 mediated genome editing in <Emphasis Type="Italic">Bacillus subtilis</Emphasis>

Genome editing using engineered nucleases has rapidly transformed from a niche technology to a mainstream method used in various host cells. Its widespread adoption has been largely developed by the emergence of the clustered regularly interspaced short palindromic repeats (CRISPR) system, which uses an easily customizable specificity RNA-guided DNA endonuclease, such as Cas9. Recently, CRISPR/Cas9 mediated genome engineering has been widely applied to model organisms, including Bacillus subtilis, enabling facile, rapid high-fidelity modification of endogenous native genes. Here, we reviewed the recent progress in B. subtilis gene editing using CRISPR/Cas9 based tools, and highlighted state-of-the-art strategies for design of CRISPR/Cas9 system. Finally, future perspectives on the use of CRISPR/Cas9 genome engineering for sequence-specific genome editing in B. subtilis are provided.     

Erlotinib inhibits colon cancer metastasis through inactivation of TrkB-dependent ERK signaling pathway

The distal metastasis is the main cause of death in patients with colon cancer. Tyrosine receptor kinase B (TrkB) and ERK signals may be the potential targets for the treatment of colon cancer metastasis. This study aims to investigate whether erlotinib inhibits distant metastasis of colon cancer by regulating TrkB and ERK signaling pathway. Human colon adenocarcinoma cell lines (SW480 and Caco-2) pretreated with exogenous C-X-C motif chemokine ligand 8 (CXCL8) were used to assess the suppressive effect of erlotinib on tumor metastasis, including anoikis, epithelial-mesenchymal transformation (EMT), migration, and invasion. Through TrkB overexpression, Akt suppression, and ERK suppression, the roles of TrkB, Akt, and ERK in erlotinib-induced metastasis inhibition of colon cancer cells were explored. The results showed that erlotinib alleviated CXCL8-induced metastasis of the colon cancer cells. Overexpression of TrkB in colon cancer cells eliminated the effect of erlotinib on anoikis, inhibition of EMT, migration, and invasion, and downregulation of p-ERK and p-Akt. Furthermore, the inhibition of ERK activation instead of Akt activation was found to participate in erlotinib-mediated metastasis resistance, including anoikis, inhibition of EMT, migration, and invasion. In conclusion, erlotinib inhibits colon cancer cell anoikis resistance, EMT, migration, and invasion by inactivating TrkB-dependent ERK signaling pathway.     

Metabolomic study of the bone trabecula of osteonecrosis femoral head patients based on UPLC–MS/MS

The severity and/or progression of osteonecrosis of the femoral head (ONFH) are commonly assessed by radiography, nuclear magnetic resonance image which aren’t invariably correlated to severity of disease and may be disturbed by other factors. Consequently, exploring the novel biochemical signatures of ONFH may be beneficial for diagnosing and understanding this disease. In this work, a bone trabecula metabolomics was undertaken to determine the expression pattern of low molecular mass metabolites in patients of femoral head necrosis based on the ultra-performance liquid chromatography/time-of-flight tandem mass spectrometry (UPLC/TOF MS/MS). Histological study showed that necrotic bone was characterized by necrosis, fibrosis and lacuna, but adjacent “normal” bone was pathologically normal. Principal component analysis in combination with orthogonal partial least-squares discrimination analysis was used to find out changed metabolites. MS/MS was used to speculate the corresponding molecule. Both osteonecrotic bone trabecula (ONBT) and adjacent “normal” bone trabecula (ANBT) showed higher levels of amino acids, such as proline, arginine, glutamine, dipeptides and lower levels of antioxidants. Most disrupted lipids, such as fatty acid esters, glycerophospholipids, sphingolipids, were found in osteonecrotic zone. The area under the receiver operating characteristic curve of combinational biomarkers (d-arginine, l-proline, l-carnitine, inosine) in ONBT and ANBT was 0.996 and 0.950, respectively. Our findings might provide a significant insight to understand the metabolic mechanism and diagnosis biomarkers of ONFH in the future.     

中分辨率遥感像元尺度生物土壤结皮覆盖与植被及土壤间的交互关系

生物土壤结皮（Biological soil crust,BSC）作为干旱、半干旱地区健康生态系统的重要组成部分,具有固碳、固氮、降低土壤水分蒸发、减少土壤径流等多种生态功能,这些功能与BSC的发展阶段及覆盖度密切相关,使其在维系荒漠自然生态系统的稳定性中扮演重要角色。因此,结合遥感数据尺度定量研究影响BSC分布的环境因素是评估沙区生态系统稳定性并以此为依据进行沙化土地治理的重要基础性研究。影响BSC覆盖度的环境因子较为复杂,现有研究方法存在两方面局限,其一是多为小尺度定性分析,其二是多侧重于孤立地分析BSC与环境因子间的单向关系。利用结构方程模型（Structural Equation Modeling,SEM）中的非递归路径分析（Nonrecursive Path Analysis,NPA）对遥感30 m分辨率像元尺度BSC覆盖度、植被覆盖度、土壤pH值、盐度、有机质与粒度进行路径分析,旨在使用综合性方法从整体上阐明大尺度BSC分布与植被、土壤间的交互关系。结果表明：（1）在毛乌素沙地,BSC覆盖度受各环境因子综合影响,无法用单一变量说明。BSC覆盖度与植被覆盖度、土壤有机质、平均粒径和细颗粒占比呈极显著正相关（P<0.01）,与粗颗粒占比呈极显著负相关（P<0.01）。（2） BSC覆盖度与植被覆盖度通过有机质相互影响,BSC覆盖度对植被覆盖度有较大的正向直接影响,路径系数（Path Coefficient,PC）=0.43（P>0.05）,植被覆盖度对BSC覆盖度有交大的负向影响（PC=－0.22;P>0.05）。（3）土壤平均粒径和细颗粒占比均正向影响BSC的覆盖度,其中,平均粒径对BSC覆盖度的总体影响较大（PC=0.67;总效果值=0.590）,细颗粒占比对BSC覆盖度的间接影响较大（间接效果值=0.052）。（4）盐度对BSC覆盖度呈显著负向直接影响（PC=－0.41;P<0.05;总效果值=－0.398）,pH值对BSC覆盖度有极小的正向影响（总效果值=0.072）。上述研究结果可为遥感探测BSC、制定有效的荒漠生态系统保护与修复政策措施提供科学依据。     

城市湿地公园的生态补水模式及其净化效果与生态效益

以常州市蔷薇湿地公园为例,提出一种“垂直流＋水平流”新型生态补水复合模式,探讨了该模式的环境效应和经济效益.结果表明：该补水模式不但能净化受污染的源水(城市河水),而且可有效提升城市湿地公园的生态景观功能.城市湿地公园内的大部分景观水体可达到全身接触性娱乐类景观功能要求,运行成本仅为常规补水工艺的5%～25%,具有较好的环境效应和经济效益.