Diazoxide Pretreatment Prevents Aβ1–42 Induced Oxidative Stress in Cholinergic Neurons Via Alleviating NOX2 Expression

The aggregation and accumulation of amyloid-β (Aβ) plays a significant role in the pathogenesis of Alzheimer’s disease. Aβ is known to increase free radical production in neuronal cells, leading to oxidative stress and cell death. Diazoxide (DZ), a highly selective drug capable of opening mitochondrial ATP-sensitive potassium channels, has neuroprotective effects against neuronal cell death. However, the mechanism through which DZ protects cholinergic neurons against Aβ-induced oxidative injury is still unclear. The present study was designed to investigate the effects of DZ pretreatment against Aβ_1–42 induced oxidative damage and cytotoxicity. Through measures of DZ effects on Aβ_1–42 induced cellular damage, reactive oxygen species (ROS) and MDA generation and expressions of gp91phox and p47phox in cholinergic neurons, new insights into the neuroprotective mechanisms can be derived. Aβ_1–42 significantly decreased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide levels and increased ROS and MDA production; all effects were attenuated by pretreatment with DZ or diphenyleneiodonium chloride (a NOX2 inhibitor). Pretreatment with DZ also attenuated the upregulation of NOX2 subunits (gp91phox and p47phox) induced by Aβ_1–42. Since NOX2 is one of the main sources of free radicals, these results suggest that DZ can counteract Aβ_1–42 induced oxidative stress and associated cell death by reducing the level of ROS and MDA, in part, by alleviating NOX2 expression.     

保护性耕作对农田碳、氮效应的影响研究进展

作物产量的高低主要取决于土壤肥力,如何保持并提高土壤肥力是确保我国粮食安全和农业可持续发展的重要任务,也是众多学者关注的焦点。土壤有机碳和氮素是评价土壤质量的重要指标,其动态平衡直接影响土壤肥力和作物产量。随着全球气候变化及环境污染问题的愈加突出,农田土壤固碳及提高氮效率成为各界科学家研究的热点。目前,保护性耕作已成为发展可持续农业的重要技术之一,对土壤固碳及氮素的利用具有很大的影响。深入了解保护性耕作对土壤有机碳固持与氮素利用效率提高的影响机制,对于正确评价土壤肥力有着重要意义。但由于气候、土壤及种植制度等条件不一致,关于保护性耕作对农田碳、氮效应结论不一。阐述了国际上保护性耕作对农田系统土壤有机碳含量变化及其分解排放(如CO₂和CH₄)、氮素变化及其矿化损失(如NH₃挥发、N₂O排放与氮淋失)和碳氮素相互关系(如C/N层化率)影响的研究进展,并分析了其影响因素和相关机理。尽管国内保护性耕作的研究已进行30 多年,但在土壤有机碳与氮素方面与国外相比依然有较大的差距。保护性耕作对土壤固碳与氮素利用的影响机制,碳素和氮素在土壤-植株-大气系统中的转移变化,及结合农事管理等综合评价其生态效应的研究很少。在此基础上,提出未来我国保护性耕作在土壤有机碳固定和氮素利用方面的重点研究方向:(1)在定位试验基础上进一步探讨保护性耕作对土壤有机碳及氮素利用的影响机制;(2)深入研究土壤有机碳和氮素的相互关系及其对土壤肥力的影响;(3)结合环境保护与土壤可持续管理对保护性耕作农田土壤固碳及氮素高效利用的系统评价研究;(4)加强保护性耕作对农田碳、氮效应的宏观研究,合理评价保护性耕措施下对农田碳、氮综合效应。     

Structure of the ribosome associating GTPase HflX

The HflX‐family is a widely distributed but poorly characterized family of translation factor‐related guanosine triphosphatases (GTPases) that interact with the large ribosomal subunit. This study describes the crystal structure of HflX from Sulfolobus solfataricus solved to 2.0‐Å resolution in apo‐ and GDP‐bound forms. The enzyme displays a two‐domain architecture with a novel “HflX domain” at the N‐terminus, and a classical G‐domain at the C‐terminus. The HflX domain is composed of a four‐stranded parallel β‐sheet flanked by two α‐helices on either side, and an anti‐parallel coiled coil of two long α‐helices that lead to the G‐domain. The cleft between the two domains accommodates the nucleotide binding site as well as the switch II region, which mediates interactions between the two domains. Conformational changes of the switch regions are therefore anticipated to reposition the HflX‐domain upon GTP‐binding. Slow GTPase activity has been confirmed, with an HflX domain deletion mutant exhibiting a 24‐fold enhanced turnover rate, suggesting a regulatory role for the HflX domain. The conserved positively charged surface patches of the HflX‐domain may mediate interaction with the large ribosomal subunit. The present study provides a structural basis to uncover the functional role of this GTPases family whose function is largely unknown. Proteins 2010. © 2009 Wiley‐Liss, Inc.     

人Leptin基因在大肠杆菌中的高效表达、纯化及其生物学活性研究

将人Leptin表达质粒pBV220-OB转化E.coliJM109,经热诱导获得了目的蛋白的表达。经SDS-PAGE鉴定分析,表达产物以包涵体形式存在,目的蛋白表达量占菌体总蛋白的40%以上。通过包涵体分离,Sephacryl S200HR凝胶和DEAE52离子交换层析及Hypersil C18柱反相色谱纯化,获得纯度在95%以上,内毒素含量小于10EU/mg的高纯度的重组人Leptin。Western-blot鉴定表明,纯化表达产物能和抗Leptin抗体特异性结合;蛋白质N端15个氨基酸序列分析结果和预期的序列一致。纯化产物经复性处理,其分子中Cys96和Cys146形成二硫键。体内活性检测显示,纯化和复性的rh-Leptin明显抑制BALB/c小鼠的进食和体重增长,提示其具有明显的生物学活性。     

Quality,bioactivity study,and preclinical acute toxicity,safety pharmacology evaluation of PEGylated recombinant human endostatin (M2ES)

Endostar, a potent endogenous antiangiogenic factor, is wildly used in clinics. However, it was easily degraded by enzymes and rapidly cleared by the kidneys. To overcome these shortcomings, PEGylated recombinant human endostatin was developed. In this study, the purity of M₂ES was evaluated by silver stain and reversed‐phase high‐performance liquid chromatography. Ultraviolet spectrum was used to examine the structural of M₂ES and endostar. The bioactivity and antitumor efficacy of M₂ES were evaluated using an in vitro endothelial cell migration model and athymic nude mouse xenograft model of a heterogeneous lung adenocarcinoma, respectively. A preclinical study was performed to evaluate the acute toxicity and safety pharmacology in rhesus monkeys. The purity of M₂ES was more than 98%; PEG modification has no effect on endostatin structure. Compared with the control group, M₂ES dramatically retards endothelial cell migration and tumor growth. After intravenous (IV) infusions of M₂ES at a dose level of three and 75 mg/kg in rhesus monkeys, there was no observable serious adverse event in both acute toxicity and safety pharmacology study. On the basis of the quality and bioactivity study data of M₂ES and the absence of serious side effect in rhesus monkeys, M₂ES was authorized to initiate a phase I clinical trial.     

“缺体回交法”选育普通小麦—山羊草异代换系的研究

利用从兰单体自交分离得到的5个自花结实的4D缺体小麦(映72180、块天选15等)作母本与11个山羊草(Ae.speltoides, Ae.sharonensis等)杂交,再以4D缺体为轮回亲本对杂种进行回交,借助于幼胚培养技术,获得了缺天选15×拟斯卑尔脱山羊草二体异代换系,缺72180×沙融山羊草单体异代换系。代换系生长发育良好,育性基本正常,表明山羊草的4S染色体能够补偿小麦缺失的4D染色体的功能。证明利用“缺体回交法”选育普通小麦—山羊草异代换系是有效的和可行的。     

西南喀斯特木本植物区系成分的纬度变异格局

本研究选取3个极具代表性的喀斯特原生植被区域,即北热带的广西弄岗自然保护区、南亚热带的广西龙虎山自然保护区和中亚热带的贵州茂兰自然保护区为研究对象,通过野外实地调查及查阅文献,整理出3个区域的木本植物名录,比较分析木本植物种类组成和区系成分的纬度变异格局。结果显示,弄岗、龙虎山和茂兰3个不同气候区域内的木本植物分别有104科425属936种、99科363属636种和100科352属995种。木本植物区系均以热带亚热带成分为主,其优势科热带性质随纬度升高逐步降低。热带区系成分与温带区系成分的比值(R/T)反映出,弄岗和龙虎山自然保护区的木本植物区系属的热带性质明显,而茂兰自然保护区植物属的热带性质较弱,温带性质属的比重随纬度升高而增加。科、属、种相似性系数表明,弄岗和龙虎山间的木本植物亲缘关系较密切,弄岗和茂兰间次之,而龙虎山和茂兰间亲缘关系较远。     

新疆伊犁哈萨克族的体质特征研究

作者对新疆伊犁地区的哈萨克族大、中专学生进行了活体观察和测量,观察30项（551名,男262,女289）测量92项（404人,男226,女178）,所得数据进行了统计学处理和分析,计算了头面部和体部的31项指数,并进行了指数分型处理。     

普通小麦：华山新麦草异代换系的选育及细胞遗传学研究

利用缺体小麦－华山新麦草七倍体杂种（２ｎ＝４９,ＡＡＢＢＤＤＮ）杂交,Ｆ１再瑟相应的缺体小麦回交２次,在ＢＣ２Ｆ１镜检选出２ｎ＝４１的植株,同时套代自交,选育出５Ａ和３Ｄ两种异代换系。单体找换植侏自交产生二体代换植侏的频率为２３．１６％。５Ａ代换植侏在减数分裂中期Ⅰ２１Ⅱ的出现频率平均为８４．５２％,３Ｄ代换植侏２１Ⅱ的出现频率平均为６２．６１％。异代换系均生长旺盛,结实正常,说明异染色体能较好地     

Analysis of gamma c-family cytokine target genes. Identification of dual-specificity phosphatase 5 (DUSP5) as a regulator of mitogen-activated protein kinase activity in interleukin-2 signaling

Interleukin (IL)-2, IL-4, IL-7, IL-9, IL-15, and IL-21 form a family of cytokines based on their sharing the common cytokine receptor gamma chain, gamma(c), which is mutated in X-linked severe combined immunodeficiency (SCID). As a step toward further elucidating the mechanism of action of these cytokines in T-cell biology, we compared the gene expression profiles of IL-2, IL-4, IL-7, and IL-15 in T cells using cDNA microarrays. IL-2, IL-7, and IL-15 each induced a highly similar set of genes, whereas IL-4 induced distinct genes correlating with differential STAT protein activation by this cytokine. One gene induced by IL-2, IL-7, and IL-15 but not IL-4 was dual-specificity phosphatase 5 (DUSP5). In IL-2-dependent CTLL-2 cells, we show that IL-2-induced ERK-1/2 activity was inhibited by wild type DUSP5 but markedly increased by an inactive form of DUSP5, suggesting a negative feedback role for DUSP5 in IL-2 signaling. Our findings provide insights into the shared versus distinctive actions by different members of the gamma(c) family of cytokines. Moreover, we have identified a DUSP5-dependent negative regulatory pathway for MAPK activity in T cells.