Evidence that NF-κB and MAPK Signaling Promotes NLRP Inflammasome Activation in Neurons Following Ischemic Stroke

Multi-protein complexes, termed “inflammasomes,” are known to contribute to neuronal cell death and brain injury following ischemic stroke. Ischemic stroke increases the expression and activation of nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) Pyrin domain containing 1 and 3 (NLRP1 and NLRP3) inflammasome proteins and both interleukin (IL)-1β and IL-18 in neurons. In this study, we provide evidence that activation of either the NF-κB and MAPK signaling pathways was partly responsible for inducing the expression and activation of NLRP1 and NLRP3 inflammasome proteins and that these effects can be attenuated using pharmacological inhibitors of these two pathways in neurons and brain tissue under in vitro and in vivo ischemic conditions, respectively. Moreover, these findings provided supporting evidence that treatment with intravenous immunoglobulin (IVIg) preparation can reduce activation of the NF-κB and MAPK signaling pathways resulting in decreased expression and activation of NLRP1 and NLRP3 inflammasomes, as well as increasing expression of anti-apoptotic proteins, Bcl-2 and Bcl-xL, in primary cortical neurons and/or cerebral tissue under in vitro and in vivo ischemic conditions. In summary, these results provide compelling evidence that both the NF-κB and MAPK signaling pathways play a pivotal role in regulating the expression and activation of NLRP1 and NLRP3 inflammasomes in primary cortical neurons and brain tissue under ischemic conditions. In addition, treatment with IVIg preparation decreased the activation of the NF-κB and MAPK signaling pathways, and thus attenuated the expression and activation of NLRP1 and NLRP3 inflammasomes in primary cortical neurons under ischemic conditions. Hence, these findings suggest that therapeutic interventions that target inflammasome activation in neurons may provide new opportunities in the future treatment of ischemic stroke.     

干旱胁迫和正常灌溉条件下玉米产量性状的QTL分析

产量及其产量因子是衡量玉米耐旱能力的重要性状。本研究利用Lo1067×Y i72的F2∶3家系进行产量性状的数量性状位点(QTL)的分析。结果表明,在正常水分条件和开花期干旱胁迫条件下,分别有14个QTL与产量性状穗重、粒重、轴重、百粒重、穗数、穗粒数有关。此外,还检测到7个与抗旱指数(TI)相关的QTL。各QTL所解释的表型变异在1%~78%;这些QTL以部分显性和超显性为主。不同胁迫条件下检测到的QTL不一致,说明存在显著的QTL与环境互作。     

Theoretical Insights into Catalytic Mechanism of Protein Arginine Methyltransferase 1

Protein arginine methyltransferase 1 (PRMT1), the major arginine asymmetric dimethylation enzyme in mammals, is emerging as a potential drug target for cancer and cardiovascular disease. Understanding the catalytic mechanism of PRMT1 will facilitate inhibitor design. However, detailed mechanisms of the methyl transfer process and substrate deprotonation of PRMT1 remain unclear. In this study, we present a theoretical study on PRMT1 catalyzed arginine dimethylation by employing molecular dynamics (MD) simulation and quantum mechanics/molecular mechanics (QM/MM) calculation. Ternary complex models, composed of PRMT1, peptide substrate, and S-adenosyl-methionine (AdoMet) as cofactor, were constructed and verified by 30-ns MD simulation. The snapshots selected from the MD trajectory were applied for the QM/MM calculation. The typical S_N2-favored transition states of the first and second methyl transfers were identified from the potential energy profile. Deprotonation of substrate arginine occurs immediately after methyl transfer, and the carboxylate group of E144 acts as proton acceptor. Furthermore, natural bond orbital analysis and electrostatic potential calculation showed that E144 facilitates the charge redistribution during the reaction and reduces the energy barrier. In this study, we propose the detailed mechanism of PRMT1-catalyzed asymmetric dimethylation, which increases insight on the small-molecule effectors design, and enables further investigations into the physiological function of this family.     

土族生物资源利用相关的传统知识多样性

生物资源利用相关的传统知识是对生物资源进行识别和利用的传统知识系统。随着现代生物技术的发展, 这类传统知识显示出其在科学、经济、文化乃至粮食安全战略方面的价值。本研究根据《生物多样性相关传统知识分类、调查与编目技术规定(试行)》, 对我国青海省土族聚集区的土族生物资源利用相关的传统知识进行了系统调查与编目, 并借鉴生物多样性测度方法, 创建了传统知识多样性指数计算方法, 对土族生物资源利用相关传统知识多样性进行分析。结果如下: (1)编目现存的土族生物资源利用相关的传统知识词条共424条; (2)土族传统知识的α多样性指数D_TK = 0.67, 表明其传统知识多样性较高; 土族传统知识的β_wtk多样性指数在不同的县域差异较大, 表明其在县域之间存在差异, 在空间上分布不连续、不均匀。本研究利用生物多样性指数验证了传统知识的定量研究方法, 说明传统知识多样性指数不仅可用于定量研究表征区域传统知识的多样性, 揭示不同空间区域内的分布特征, 还可为未来构建传统知识的定量研究体系提供重要的参考依据。     

幼龄小白鼠全脑5SrRNA的全核苷酸序列测定

<正> 5S rRNA是一种稳定的独立小分子,在原核和真核细胞中都牢固地结合在核糖体的大亚基上。它在蛋白质生物合成过程中具有重要作用。微生物、大鼠肝细胞及某些植物细胞5S rRNA的一级结构都已经测定,脑细胞5SrRNA的一级结构尚未见报道。因此,我们对幼龄小鼠全脑5S rRNA的序列进行了分析。我们用辜祥荣等人的方法分离和纯化5S rRNA,5S rRNA3′-末端标记参照Peattie的方法(2)。3′-末端标记5S rRNA的序列分析用化学降解-凝胶直读法及特异RNa8e降解直     

DNA vaccination against foot-and-mouth disease via electroporation: study of molecular approaches for enhancing VP1 antigenicity

BACKGROUND: Foot-and-mouth disease virus (FMDV) affects susceptible livestock animals and causes disastrous economic impact. Immunization with plasmid expressing VP1 that contains the major antigenic epitope(s) of FMDV as cytoplasmic protein (cVP1) failed to elicit full protection against FMDV challenge. MATERIALS AND METHODS: In this study, mice were immunized via electroporation with four cDNA expression vectors that were constructed to express VP1 of FMDV, as cytoplasmic (cVP1), secreted (sVP1), membrane-anchored (mVP1) or capsid precursor protein (P1), respectively, to evaluate whether expression of VP1 in specific subcellular compartment(s) would result in better immune responses. RESULTS: Electroporation enhanced immune responses to vectors expressing cVP1 or P1 and expedited the immune responses to vectors expressing sVP1 or mVP1. Immunization of mice via electroporation with mVP1 cDNA was better than sVP1 or cVP1 cDNA in eliciting neutralizing antibodies and viral clearance protection. Vaccination with P1 cDNA, nonetheless, yielded the best immune responses and protection among all four cDNAs that we tested. CONCLUSIONS: These results suggest that the antigenicity of a VP1 DNA vaccine can be significantly enhanced by altering the cellular localization of the VP1 antigen. Electroporation is a useful tool for enhancing the immune responses of vectors expressing VP1 or P1. By mimicking FMDV more closely than that of transgenic VP1 and eliciting immune responses favorably toward Th2, transgenic P1 may induce more neutralizing antibodies and better protection against FMDV challenge.     

对TMV不同抗性番茄品种的叶绿体DNA限制性内切酶酶谱分析

选用对TMV有抗性和敏感的番茄品种、制备其ct-DNA, 用限制性内切酶BumHI、EcoRI和PstI完全酶解, 三种酶切图谱与前人报道一致, 由酶切片段计算番茄ct-DNA。分子量约为156.9kb。比较抗性和敏感品种的ct-DNA图谱, 发现三种酶切图谱均存在差异, 但由差异片段计算分子量之和又很除近。我们推测这是由于检基顺序变异或小段DNA顺序插入或缺失所造成, 由此证明, 叶绿体基因组与核中的TMV抗性基因, 共同决定着植物体对TMV的抗性。     

Calcium regulation of growth and differentiation of mouse epidermal cells in culture

Modification of the ionic calcium concentration in the culture medium markedly alters the pattern of proliferation and differentiation in cultured mouse epidermal cells. When medium calcium is lowered to 0.05--0.1 mM, keratinocytes proliferate rapidly with a high growth fraction and do not stratify, but continue to synthesize keratin. The cells grow as a monolayer for several months and can be subcultured and cloned in low Ca++ medium. Ultrastructural examination of cells cultured under low Ca++ conditions reveals widened intercellular spaces, abundant microvilli and perinuclear organization of tonofilaments and cellular organelles. Desmosomes are absent. Epidermal cells growing as a monolayer in low Ca++ can be induced to terminally differentiate by adding calcium to the level normally found in the culture medium (1.2 mM). Cell-to-cell contact occurs rapidly and desmosomes form within 2 hr. The cells stratify by 1--2 days and terminally differentiate with cell sloughing by 3--4 days. After Ca++ addition, DNA synthesis decreases with a lag of 5--10 hr and is totally inhibited within 34 hr. In contrast, RNA and protein synthesis continue at 40--50% of the low Ca++ level at day 3, a time when many cells are detaching from the culture dish. Keratin synthesis is unaffected by the Ca++ switch.