Stereoselective metabolism of fenoxaprop‐ethyl and its chiral metabolite fenoxaprop in rabbits

The stereoselective metabolism of the enantiomers of fenoxaprop‐ethyl (FE) and its primary chiral metabolite fenoxaprop (FA) in rabbits in vivo and in vitro was studied based on a validated chiral high‐performance liquid chromatography method. The information of in vivo metabolism was obtained by intravenous administration of racemic FE, racemic FA, and optically pure (−)‐(S)‐FE and (+)‐(R)‐FE separately. The results showed that FE degraded very fast to the metabolite FA, which was then metabolized in a stereoselective way in vivo: (−)‐(S)‐FA degraded faster in plasma, heart, lung, liver, kidney, and bile than its antipode. Moreover, a conversion of (−)‐(S)‐FA to (+)‐(R)‐FA in plasma was found after injection of optically pure (−)‐(S)‐ and (+)‐(R)‐FE separately. Either enantiomers were not detected in brain, spleen, muscle, and fat. Plasma concentration–time curves were best described by an open three‐compartment model, and the toxicokinetic parameters of the two enantiomers were significantly different. Different metabolism behaviors were observed in the degradations of FE and FA in the plasma and liver microsomes in vitro, which were helpful for understanding the stereoselective mechanism. This work suggested the stereoselective behaviors of chiral pollutants, and their chiral metabolites in environment should be taken into account for an accurate risk assessment. Chirality, 2011. © 2011 Wiley‐Liss, Inc.     

Effects of daintain/AIF-1 on β cell dysfunction in INS-1 cells

We investigated the effects of daintain/AIF-1, a novel inflammatory cytokine, on INS-1β cells. Cells incubated with daintain/AIF-1 showed decreased cell viability and glucose-stimulated insulin secretion, as well as upregulated apoptosis and NO production. These deleterious effects of daintain/AIF-1 indicate that daintain/AIF-1 plays important roles in the dysfunction of pancreatic β cells in type-1 diabetes.     

一株有色金属矿山1-亚硝基-2-萘酚降解菌的筛选、鉴定和降解特性

1-亚硝基-2-萘酚是一类有色选矿药剂代表性的新型浮选药剂,在采选冶行业为了提高低品位矿物资源的利用率,被大量投入使用,该药剂的高稳定性进一步增大了矿山环境中重金属与有机选冶药剂复合污染的治理难度。微生物修复作为稠环芳烃(polycyclic aromatic hydrocarbons,PAHs)类污染物重要技术手段之一,具有安全、经济、高效和无二次污染等特点。【目的】本研究从我国广西河池市周边的典型有色金属尾矿环境中分离出1株高效降解1-亚硝基-2-萘酚的菌株,并分析其降解特性及其潜在的代谢途径,从而探究矿山复合污染生态系统中稠环芳烃类污染物的微生物修复技术的应用前景。【方法】从有色金属尾矿样本中筛选到以1-亚硝基-2-萘酚为唯一碳源的菌株,经16S rRNA基因序列鉴定,结合气相色谱-质谱联合(gas chromatography-mass spectrometer,GC-MS)检测分析菌株对1-亚硝基-2-萘酚的降解特性及中间代谢产物,并推测可能的代谢途径。【结果】筛选获得1株高效降解1-亚硝基-2-萘酚的Pseudomonas putida CUGB-JL11菌株,经鉴定为革兰氏阴性杆的恶臭假单胞菌。该菌株最适温度为30 ℃左右,最适pH值范围为6-8条件下,菌株5 d内对40 mg/L 1-亚硝基-2-萘酚的降解率高达81%。降解中间代谢产物主要为苯环类物质：苯甲羟肟酸甲酯和苯丙胺,但其母体及大部分中间产物都降解为小分子物质或者被完全降解。【结论】恶臭假单胞菌P. putida CUGB-JL11具有良好的1-亚硝基-2-萘酚降解能力和较强的环境适应性,有进一步被开发为微生物菌剂以用于稠环芳烃类污染修复的巨大潜力,为有色金属矿山生态系统中重金属和有机选冶药剂复合污染的微生物修复研究提供了理论依据和可利用的微生物资源。     

长江中下游平原区生态保护红线的划定——以江苏省为例

科学划定生态保护红线（Ecological Protection Red Line,EPRL）并严格保护是中国政府建设生态文明的重要战略举措,对国土空间可持续发展具有重要意义。通过解析生态保护红线制度的内涵、目标与划定标准提出生态保护红线的4项基础性问题。针对长江中下游平原面临的生物多样性减少、生境斑块破碎化以及农业与生态空间冲突等问题,考虑生态保护红线的区域差异性,以江苏省为研究区,结合区域特征利用生境维持、气候调节、水源涵养、粮食供给、土壤保持5项生态系统服务功能,基于生态安全格局研究框架探讨了相应的生态保护红线划定方法。研究结果表明：①江苏省生态源地主要由区域内河流、湖泊、滩涂与林地构成,各项生态系统服务功能空间格局差异化显著,限于农田覆盖范围广泛与水系网络结构发达,源地斑块破碎化现象严重。②江苏生态保护红线主要由湿地、林草地、农用地等地类构成,红线核心区以湿地为主,廊道与节点则以农用地为主。生态保护红线可根据源地景观规模分为3级,其中I级EPRL占比达72.87%,可基本代表核心红线图斑。③生态廊道与生态节点的设置有助于提高生境图斑的连通性,江苏省生态廊道共47条,包括33条关键生态廊道与14条潜在生态廊道,总长度分别为1296.27km、1726.33km。研究结果可为长江中下游平原的生态保护红线划定创新思路,并希冀对生态保护红线制度约束下的国土空间管控与管制制度改革具有借鉴价值。     

肝细胞癌患者自噬相关基因的预后作用

构建由自噬相关基因组成的预后模型,预测肝细胞癌(HCC)患者的生存预后情况,为其个性化诊疗和临床研究提供依据。利用TCGA数据库中HCC的测序信息与人类自噬数据库联合,筛选差异表达的自噬相关基因,对其进行GO富集与KEGG通路分析;通过单因素与多因素Cox分析筛选与患者生存预后明显相关的风险基因,构建预后风险评分模型;根据模型计算患者风险值并验证模型,利用GEPIA2.0网页工具与HPA数据库对风险基因在HCC中的表达情况以及与生存预后的关系进行验证。结果发现,HCC肿瘤组织相较正常组织共筛选到61个差异表达的自噬相关基因(表达上调57个,下调4个),GO富集与KEGG通路分析显示均与自噬有关;单因素Cox分析共筛选到12个与患者生存预后相关的基因,多因素Cox分析后共有4个基因被纳入预后风险评分模型,分别是SQSTM1、HDAC1、RHEB和ATIC,计算公式为:风险值(risk score)=SQSTM1表达量×0.185+HDAC1表达量×0.382+RHEB表达量×0.423+ATIC表达量×0.438;K-M生存曲线显示高风险组生存率低于低风险组,风险曲线提示4个基因与不良预后密切相关,ROC曲线证明模型具有预测意义;GEPIA2.0网页工具以及HPA数据库表明高表达4个基因均导致患者生存率降低。所构建预后风险评分模型可有效预测HCC患者生存预后情况,并提供个性化诊疗策略。     

薏苡黑穗病菌的宏基因组测序及分类解析

旨在寻找薏苡黑穗病害的关键致病菌群,指导薏苡的黑穗病害防治.本研究利用ITS高通量测序技术对薏苡的黑穗病瘿的真菌进行检测,解析薏苡黑穗病瘿组织的菌群组成和群落多样性.结果表明,薏苡黑穗病瘿中存在多种真菌菌群,总共聚类得到2 699条物种OTU,所有样品共同拥有94条,总共注释得到9门、19纲、37目、68科和85属.在属分类水平发现2个与黑穗病相关的近缘真菌属Sporisorium和Ustilago,其中Sporisorium为优势菌群.3份样品Shannon多样性指数物种大小为XY-2＞XY-3＞XY-1,且XY-1和XY-3的菌群组成更为相似.本研究结果对于薏苡黑穗病菌的分离鉴定和绿色防控具有重要指导意义.     

镰刀菌Q7-31T内切葡聚糖酶Egn20的分离纯化鉴定及酶学特性

摘要:【目的】从镰刀菌Q7-31T燕麦秸秆诱导发酵的粗酶液中分离、纯化并鉴定内切葡聚糖酶,研究其酶学特性。为丰富和完善镰刀菌的酶系信息提供理论支持。【方法】以燕麦秸秆为碳源诱导发酵培养菌株,采用Sephacry S-100凝胶柱层析和DEAE琼脂糖弱阴离子交换柱层析对粗酶液进行分离纯化得到内切葡聚糖酶Egn20,随后对其进行了酶学性质分析和串联质谱鉴定。【结果】分离纯化得到内切葡聚糖酶Egn20,其分子量为55.37 kDa,等电点为7.44;酶学特性结果表明:Egn20对羧甲基纤维素的最适反应温度为40 ℃,最适pH为6.0,该酶在45 ℃和弱酸性环境下较稳定,Fe2+对其有激活作用,Na+、Ca2+、Mg2+、Zn2+和K+抑制该酶活性,Hg2+使该酶失活;酶学特性和串联质谱鉴定的结果表明Egn20属于GH7家族。【结论】从镰刀菌Q7-31T粗酶液中分离纯化得到内切葡聚糖酶Egn20,并对其进行了酶学性质的研究和串联质谱鉴定,结果表明Egn20为GH7家族内切葡聚糖酶。本研究为丰富和完善镰刀菌的酶系信息提供了理论和数据支持。     

Interleukin-35 Inhibits Endothelial Cell Activation by Suppressing MAPK-AP-1 Pathway

Vascular response is an essential pathological mechanism underlying various inflammatory diseases. This study determines whether IL-35, a novel responsive anti-inflammatory cytokine, inhibits vascular response in acute inflammation. Using a mouse model of LPS-induced acute inflammation and plasma samples from sepsis patients, we found that IL-35 was induced in the plasma of mice after LPS injection as well as in the plasma of sepsis patients. In addition, IL-35 decreased LPS-induced proinflammatory cytokines and chemokines in the plasma of mice. Furthermore, IL-35 inhibited leukocyte adhesion to the endothelium in the vessels of lung and cremaster muscle and decreased the numbers of inflammatory cells in bronchoalveolar lavage fluid. Mechanistically, IL-35 inhibited the LPS-induced up-regulation of endothelial cell (EC) adhesion molecule VCAM-1 through IL-35 receptors gp130 and IL-12Rβ2 via inhibition of the MAPK-activator protein-1 (AP-1) signaling pathway. We also found that IL-27, which shares the EBI3 subunit with IL-35, promoted LPS-induced VCAM-1 in human aortic ECs and that EBI3-deficient mice had similar vascular response to LPS when compared with that of WT mice. These results demonstrated for the first time that inflammation-induced IL-35 inhibits LPS-induced EC activation by suppressing MAPK-AP1-mediated VCAM-1 expression and attenuates LPS-induced secretion of proinflammatory cytokines/chemokines. Our results provide insight into the control of vascular inflammation by IL-35 and suggest that IL-35 is an attractive novel therapeutic reagent for sepsis and cardiovascular diseases.     

Interaction mapping between Saccharomyces cerevisiae Smc5 and SUMO E3 ligase Mms21

The multisubunit Smc5-Smc6 holocomplex (Smc5/6) plays a critical role in chromosome stability maintenance, DNA replication, homologous recombination, and double-stranded DNA damage repair. Smc5 and Smc6 form the core of the holocomplex, along with six non-SMC elements, for which most functions are not yet understood. Mms21 (Nse2), the relatively well-studied subunit in Smc5/6, contains a SP-like-RING finger motif on the C-terminus and was identified as a SUMO E3 ligase. Deletion of Mms21 is lethal; however, while deficient in DNA damage repair, SUMO ligase mutants remain viable. These functions of Mms21 in Smc5/6 are hard to address without understanding the interaction between Smc5 and Mms21. Previously, we systematically examined the architecture of Saccharomyces cerevisiae Smc5/6 and, using yeast two-hybrid methods, found that Mms21 interacts with the coiled-coil of Smc5. Later, crystallographic studies revealed the molecular arrangement of Mms21 with Smc5/6. For this study, we use a combination of limited proteolysis, mass spectrometry, and N-terminal sequencing to precisely define the interaction region of Smc5 with Mms21. In addition, using isothermal titration calorimetry, we find that Mms21 interacts with Smc5 in a 1:1 ratio with a K(d) of 0.68 μM. This combination of methods would be useful in examining the structure of any large multiprotein complex.