Puerarin attenuates endothelial insulin resistance through inhibition of inflammatory response in an IKKβ/IRS-1-dependent manner

Puerarin is an isoflavonoid isolated from the root of the plant Pueraria lobata and has been used as a prescribed drug in China for the treatment of cardiovascular diseases in the clinical practice. Puerarin possesses potential therapeutic activities for metabolic and cardiovascular disorders. However, little is yet known about its bioprotection against endothelial dysfunction insulin resistance involved. In this study, we established insulin resistance by palmitate stimulation in the endothelium and investigated the action of puerarin on the modulation of insulin sensitivity under the insulin resistant condition. Palmitate stimulation impaired insulin-mediated vasodilation in the rat aorta and puerarin treatment effectively restored the impaired vasodilation in a concentration-dependent manner (1, 10 and 50 μM). Palmitate stimulation evoked inflammatory response in endothelial cells. Puerarin inhibited IKKβ/NF-κB activation and decreased TNF-α and IL-6 production with the downregulation of relative gene overexpression. Palmitate stimulation impaired the insulin PI3K signaling pathway and reduced insulin-mediated NO production in endothelial cells. Puerarin attenuated PA-induced phosphorylation of insulin receptor substrate-1 (IRS-1) at S307 and effectively ameliorated insulin-mediated tyrosine phosphorylation of IRS-1. The beneficial modification of serine/tyrosine phosphorylation of IRS-1 restored downstream Akt/eNOS activation, and thereby increased insulin-mediated NO production. These results suggest that puerarin inhibits inflammation and attenuates endothelial insulin resistance in an IKKβ/IRS-1-dependent manner.     

Suppression of Gαs synthesis by simvastatin treatment of vascular endothelial cells

These studies explore the effects of statins on cyclic AMP-modulated signaling pathways in vascular endothelial cells. We previously observed (Kou, R., Sartoretto, J., and Michel, T. (2009) J. Biol. Chem. 284, 14734-14743) that simvastatin treatment of endothelial cells leads to a marked decrease in PKA-modulated phosphorylation of the protein VASP. Here we show that long-term treatment of mice with simvastatin attenuates the vasorelaxation response to the β-adrenergic agonist isoproterenol, without affecting endothelin-induced vasoconstriction or carbachol-induced vasorelaxation. We found that statin treatment of endothelial cells dose-dependently inhibits PKA activation as assessed by analyses of serine 157 VASP phosphorylation as well as Epac-mediated Rap1 activation. These effects of simvastatin are completely reversed by mevalonate and by geranylgeranyl pyrophosphate, implicating geranylgeranylation as a critical determinant of the stain response. We used biochemical approaches as well as fluorescence resonance energy transfer (FRET) methods with a cAMP biosensor to show that simvastatin treatment of endothelial cells markedly inhibits cAMP accumulation in response to epinephrine. Importantly, simvastatin treatment significantly decreases Gα(s) abundance, without affecting other Gα subunits. Simvastatin treatment does not influence Gα(s) protein stability, and paradoxically increases the abundance of Gα(s) mRNA. Finally, we found that simvastatin treatment inhibits Gα(s) translation mediated by Akt/mTOR/eIF4/4EBP. Taken together, these findings establish a novel mechanism by which simvastatin modulates β-adrenergic signaling in vascular wall, and may have implications for cardiovascular therapeutics.     

植被、海拔、人为干扰对大中型野生动物分布的影响——以九寨沟自然保护区为例

利用九寨沟自然保护区内8条动物监测样线在2003—2010年的生态监测数据,分析了植被、海拔、人为干扰对大中型野生动物分布的影响,结果显示:1)本监测共记录到保护区内大中型野生动物26种,其中兽类18种,鸟类8种,属于我国Ⅰ级、Ⅱ级保护动物的分别有5种和11种。2)针阔混交林和针叶林是大中型野生动物种类最丰富的两种植被类型(分别有26种和17种),阔叶林和灌丛次之(分别为10种和12种),草地最少(2种)。3)海拔2400 m以下的地方,大中型野生动物种类稀少,为10种,2400—2599 m海拔段物种数增加至20种,之后随海拔上升物种丰富度下降。4)人为干扰显著影响大中型野生动物的空间分布:历史上的人类活动将原生森林植被转化为次生的落叶阔叶林、灌丛和草地,使树正、亚纳和尖盘等地下坡位(相对海拔0—199 m)的大中型野生动物的物种丰富度降低(分别为4、2、2种)。包括旅游活动和交通在内的人为干扰,可能导致附近50m海拔范围内保护动物的缺失,并使下坡位的某些物种向高海拔处移动。生境破碎化则使野生动物的种类组成发生改变,使原有森林内部优势种,如羚牛(Budorcas taxicolor),逐渐被适应能力强的物种,如豹猫(Prionailurus bengalensis)、雉鸡(Phasianuscolchicus)和野猪(Sus scrofa)等所取代,并将长期影响野生动物种群的存活。当前九寨沟自然保护区大中型野生动物的分布是植被、海拔与人为干扰综合作用的结果。     

Construction and characterization of a high-affinity humanized SM5-1 monoclonal antibody

SM5-1 is a mouse monoclonal antibody which has a high specificity for melanoma, hepatocellular carcinoma, and breast cancer, making it a promising candidate for cancer targeting therapy. We have therefore attempted to construct a humanized antibody of SM5-1 to minimize its immunogenicity for potential clinical use. Using a molecular model of SM5-1 built by computer-assisted homology modeling, framework region (FR) residues of potential importance to the antigen binding were identified. Then, a humanized version of SM5-1 was generated by transferring these mouse key FR residues onto a human framework that was selected based on homology to the mouse framework, together with the mouse complementarity-determining region (CDR) residues. This humanized antibody retained only six murine residues outside of the CDRs but was shown to possess affinity and specificity comparable to that of the parental antibody, suggesting that it might have the potential to be developed for future clinical use.     

Eradication of hepatoma and colon cancer in mice with Flt3L gene therapy in combination with 5-FU

We developed a recombinant defective adenovirus with an insert of gene encoding extracellular domain of mouse Flt3L (Ad-mFlt3L) under control of cytomegalovirus promoter to investigate the biological efficacy of Flt3L in combination with chemotherapeutical drug, 5-FU, in eliciting an effective anti-cancer immunity in mouse hepatoma and colon cancer model systems. The constructed Ad-mFlt3L efficiently infected hepatoma and colon cancer cells both in vitro and in vivo, leading to a high production of mFlt3L proteins in association with accumulation of DCs, NK cells and lymphocytes in local tumor tissues. Administration of Ad-mFlt3L can protect bone marrow injury caused by 5-Fu and stimulates proliferation and maturation of lymphocytes, APCs and NKs. Intratumoral injection of Ad-mFlt3L followed by an intraperitoneal administration of 5-Fu significantly inhibited tumor growth and cured established tumors. Adenovirus mediated Flt3L gene therapy synergies with chemotherapeutic drug, 5-Fu, in elicitation of long-lasting antitumor immunity. The tumor specific immunity can be adoptively transferred into naïve animals successfully by transfusion of CD3⁺CD8⁺ T cells from the treated mice. The data suggests that adenovirus mediated Flt3L gene therapy in combination with 5-Fu chemotherapy may open a new avenue for development of anti-cancer chemogenetherapy.     

Inhibitory Activity of Honeysuckle Extracts against Influenza A Virus In Vitro and In Vivo

Virologica Sinica - Honeysuckle has been used in the treatment of influenza virus infection for thousands of years in China. However, its main active components and the functional mechanisms remain...     

生物制品学教学改革思考

随着现代生物技术的快速发展,培养高素质生物技术型人才受到越来越广泛的关注。生物制品学课程是生物学专业应掌握的一门基础学科,但由于生物制品学是一门新兴学科,其教学内容和教学方法均处于探索阶段。文中针对生物制品学课程目前的教学现状及存在的问题,从教学内容、教学方法、考核方式等方面提出教学改革,为进一步提高生物制品学课程的教学效果、提高学生的学习兴趣打下坚实的基础。     

Establishment of an efficient seed fluorescence reporter‐assisted CRISPR/Cas9 gene editing in maize

Genome editing by clustered regularly interspaced short palindromic sequences (CRISPR)/CRISPR‐associated protein 9 (Cas9) has revolutionized functional gene analysis and genetic improvement. While reporter‐assisted CRISPR/Cas systems can greatly facilitate the selection of genome‐edited plants produced via stable transformation, this approach has not been well established in seed crops. Here, we established the seed fluorescence reporter (SFR)‐assisted CRISPR/Cas9 systems in maize (Zea mays L.), using the red fluorescent DsRED protein expressed in the endosperm (En‐SFR/Cas9), embryos (Em‐SFR/Cas9), or both tissues (Em/En‐SFR/Cas9). All three SFRs showed distinct fluorescent patterns in the seed endosperm and embryo that allowed the selection of seeds carrying the transgene of having segregated the transgene out. We describe several case studies of the implementation of En‐SFR/Cas9, Em‐SFR/Cas9, and Em/En‐ SFR/Cas9 to identify plants not harboring the genome‐editing cassette but carrying the desired mutations at target genes in single genes or in small‐scale mutant libraries, and report on the successful generation of single‐target mutants and/or mutant libraries with En‐SFR/Cas9, Em‐SFR/Cas9, and Em/En‐SFR/Cas9. SFR‐assisted genome editing may have particular value for application scenarios with a low transformation frequency and may be extended to other important monocot seed crops.     

Bryoerythrophyllum neimonggolicum X.-L.Bai & C.Feng (Pottiaceae), a new species from Inner Mongolia,China

Abstract

A new Inner Mongolian endemic species, Bryoerythrophyllum neimonggolicum X.-L.Bai & C.Feng is described and illustrated from China. It is characterized by lingulate to broad-lanceolate leaves with obtuse or rounded apex, leaf margins broadly recurved to revolute to near the apex, bulging laminal cells, basal cells with thickened walls and the absence of lax and enlarged basal cells. This species is compared with closely related species and its ecology is discussed.