副溶血性弧菌CalR调控vopB2的分子机制

【背景】副溶血性弧菌是一种非常重要的食源性致病菌,CalR蛋白是一种全局转录调节因子。III型分泌系统2 (Type 3 secretion systems 2 T3SS2)是副溶血性弧菌主要的毒力因子,vopB2是T3SS2中的一个关键效应蛋白。【目的】研究副溶血弧菌CalR对vopB2的转录调控机制。【方法】利用引物延伸实验鉴定vopB2及vtrA的转录起始位点,并根据产物的丰度判断CalR对靶基因的调控关系;采用实时定量RT-PCR研究靶基因mRNA在WT和ΔcalR中转录丰度,验证CalR对靶基因的转录调控关系,进一步利用LacZ实验通过比较β-半乳糖苷酶活性的差异来判定CalR对靶基因的调控关系;利用凝胶阻滞实验分析His-CalR对靶基因启动子区是否具有直接的结合作用。【结果】vopB2有两个转录起始位点A(-130和-28)且其活性受CalR的直接抑制;引物延伸和LacZ结果表明CalR对vtrA的转录并无调控作用。【结论】CalR直接抑制vopB2的转录,该抑制作用与vtrA无关联。     

Diallyl trisulfide induces apoptosis and inhibits proliferation of A549 cells in vitro and in vivo

Lung cancer is the leading cause of cancer-related mortality all over the world. In recent years, pulmonary adenocarcinoma has surpassed squamous cell carcinoma in frequency and is the predominant form of lung cancer in many countries. Epidemiological investigations have shown an inverse relationship between garlic (Allium sativum) consumption and death rate from many cancers. Diallyl trisulfide (DATS) is one of the garlic-derived compounds (also known as: organosulfer compounds, OSC). DATS can induce apoptosis and inhibit the growth of many cancer cell lines. Our study demonstrated that the apoptotic incidents induced by DATS were a mitochondria-dependent caspase cascade through a significant decrease of the anti-apoptotic Bcl-2 that resulted in up-regulation of the ratio of Bax/Bcl-2 and the activity of caspase-3, -8, and -9. Eventually, DATS induced the apoptosis and inhibited the proliferation in a concentration- and time-dependent manner. Furthermore, by establishing an animal model of female BALB/c nude mice with A549 xenografts, we found that oral gavage of DATS significantly retarded growth of A549 xenografts in nude mice without causing weight loss or any other side effects compared with the control group. All the evidence both in vitro and in vivo suggested that DATS could be an ideal anti-cancer drug.     

双绕蛋白质的分类与识别

蛋白质折叠识别是蛋白质结构研究的重要内容。双绕是α/β蛋白质中结构典型的常见折叠类型。选取22个家族中序列一致性小于25%的79个典型双绕蛋白质作为训练集,以RMSD为指标进行系统聚类,并对各类建立基于结构比对的概形隐马尔科夫模型(profile-HMM)。将Astral1.65中序列一致性小于95%的9 505个样本作为检验集,整体识别敏感性为93.9%,特异性为82.1%,MCC值为0.876。结果表明:对于成员较多,无法建立统一模型的折叠类型,分类建模可以实现较高准确率的识别。     

热氧老化对麦秸秆/橡胶/PE仿藤条性能的影响

秸塑复合材料(SPC)是一种使用秸秆纤维替代木材纤维的新型木塑复合材料。以麦秸秆和低密度聚乙烯为原料,利用天然橡胶增韧的特性开发出了麦秸秆/橡胶生物质仿藤条。在100℃条件下加速热氧老化60天,观察分析其力学性能和微观结构的变化规律。结果表明,初始条件下,一方面麦秸秆纤维的加入降低了仿藤条的力学性能;另一方面橡胶的加入增强了仿藤条的韧性,起到了弥补作用。老化过程中,材料表面出现裂纹,生物质和PE界面的结合官能团丧失,界面结合能力降低,力学性能下降。结合动力学模型,0~15天为快速降解阶段,材料断裂伸长率降低较快,橡胶的加入降低了老化速率,老化系数降低了70%。含有橡胶的仿藤条在15~60天的老化过程中保持较低的老化速率,起到了抗老化作用。     

GS15 forms a SNARE complex with syntaxin 5, GS28, and Ykt6 and is implicated in traffic in the early cisternae of the Golgi apparatus

The subcellular localization, interacting partners, and function of GS15, a Golgi SNARE, remain to be established. In our present study, it is revealed that unlike proteins (Bet1 and the KDEL receptor) cycling between the Golgi and the intermediate compartment (IC, inclusive of the ER exit sites), GS15 is not redistributed into the IC upon incubation at 15 degrees C or when cells are treated with brefeldin A. Immuno-electron microscopy (immuno-EM) reveals that GS15 is mainly found in the medial-cisternae of the Golgi apparatus and adjacent tubulo-vesicular elements. Coimmunoprecipitation experiments suggest that GS15 exists in a distinct SNARE complex that contains SNAREs (syntaxin5, GS28, and Ykt6) that are implicated in both ER-to-Golgi and intra-Golgi transport but not with SNAREs involved exclusively in ER-to-Golgi traffic. Furthermore, components of COPI coat can be selectively coimmunoprecipitated with GS15 from Golgi extracts. Overexpression of mutant forms of GS15 affects the normal distribution of cis- and medial-Golgi proteins (GS28, syntaxin 5, and Golgi mannosidase II), whereas proteins of the trans-Golgi and TGN (Vti1-rp2/Vti1a and syntaxin 6) and Golgi matrix/scaffold (GM130 and p115) are less affected. When the level of GS15 is reduced by duplex 21-nt small interfering RNA (siRNA)-mediated knockdown approach, diverse markers of the Golgi apparatus are redistributed into small dotty and diffuse labeling, suggesting an essential role of GS15 in the Golgi apparatus.     

吗啡对大鼠海马神经元突触传递的作用及机制探讨

目的 :从离子通道角度研究吗啡对中枢神经系统兴奋性及抑制性突触传递的作用并探讨其机制。方法 :　原代培养新生Wistar大鼠的海马神经元。采用膜片钳技术研究吗啡对其兴奋性及抑制性突触后电流及谷氨酸诱发电流的影响。结果 :①吗啡可明显增强海马神经元兴奋性突触传递 ,加吗啡后自发兴奋性突触后电流 (sEPSC)的发放频率增加了 ( 2 0 7.8± 2 0 .9) %。此作用可被阿片受体阻断剂纳洛酮阻断 (P <0 .0 1) ;②吗啡对微小兴奋性突触后电流 (mEPSC)的发放频率及谷氨酸诱发电流的幅度没有明显影响 (P >0 .0 5 ) ;③吗啡可明显抑制神经元自发抑制性突触后电流 (sIPSC) ,纳洛酮可拮抗吗啡作用 (n =13 ,P <0 .0 1)。结论 :实验结果提示吗啡对海马神经元的兴奋作用不是由于吗啡直接作用于兴奋性氨基酸—谷氨酸突触传递过程 ,而是可能由于抑制了抑制性中间神经元 ,间接产生的兴奋作用。     

Generation of <Emphasis Type="Italic">Vibrio anguillarum</Emphasis> Ghost by Coexpression of PhiX 174 Lysis <Emphasis Type="Italic">E</Emphasis> gene and Staphylococcal Nuclease <Emphasis Type="Italic">A</Emphasis> Gene

Vibrio anguillarum ghosts (VAG) were generated, for the first time, using a conjugation vector containing a ghost bacteria inducing cassette, pRK-λP_R-cI-Elysis, in which the expression of PhiX174 lysis gene E was controlled by the P _R /cI regulatory system of lambda phage. By scanning electron microscopy, holes ranging 80–200 nm in diameter were observed in the VAG. To avoid the presence of bacterial genomic DNA and an antibiotic resistance gene in the final VAG product, we constructed a new dual vector, pRK-λP_R-cI-E-SNA, containing the E-mediated lysis cassette and the staphylococcal nuclease A (SNA)-mediated DNA degradation cassette, and generated safety-enhanced VAG for use as a fish vaccine.     

Spred2 Modulates the Erythroid Differentiation Induced by Imatinib in Chronic Myeloid Leukemia Cells

Differentiation induction is currently considered as an alternative strategy for treating chronic myelogenous leukemia (CML). Our previous work has demonstrated that Sprouty-related EVH1 domainprotein2 (Spred2) was involved in imatinib mediated cytotoxicity in CML cells. However, its roles in growth and lineage differentiation of CML cells remain unknown. In this study, we found that CML CD34⁺ cells expressed lower level of Spred2 compared with normal hematopoietic progenitor cells, and adenovirus mediated restoration of Spred2 promoted the erythroid differentiation of CML cells. Imatinib could induce Spred2 expression and enhance erythroid differentiation in K562 cells. However, the imatinib induced erythroid differentiation could be blocked by Spred2 silence using lentiviral vector PLKO.1-shSpred2. Spred2 interference activated phosphorylated-ERK (p-ERK) and inhibited erythroid differentiation, while ERK inhibitor, PD98059, could restore the erythroid differentiation, suggesting Spred2 regulated the erythroid differentiation partly through ERK signaling. Furthermore, Spred2 interference partly restored p-ERK level leading to inhibition of erythroid differentiation in imatinib treated K562 cells. In conclusion, Spred2 was involved in erythroid differentiation of CML cells and participated in imatinib induced erythroid differentiation partly through ERK signaling.     

微生物育种物理诱变技术ARTP的应用进展

微生物物理诱变育种技术广泛应用于改善微生物菌种特性、提高微生物产品产量与质量,并且在生物燃料和生物修复方面也具有重要的应用价值。本文重点介绍常压室温等离子体(ARTP)物理诱变技术在微生物诱变育种方面的应用,其对具有重要工农业应用价值的产芽孢菌种具有明显的诱变优越性。进一步分析了微生物物理诱变育种技术未来的发展趋势和重要应用前景,为微生物物理诱变育种工作提供借鉴。