Possible dynamic anchor points in a benzoxazinone derivative–human oxytocin receptor system — a molecular docking and dynamics calculation

In this study, we performed a molecular docking and dynamics simulation for a benzoxazinone–human oxytocin receptor system to determine the possible hydrophobic and electrostatic interaction points in the dynamic complex. After the homology modeling, the ligand was docked into the putative active using AutoDock 3.05. After the application of energetic and structural filters, the complexes obtained were further refined with a simulated annealing protocol (AMBER8) to remove steric clashes. Three complexes were selected for subjection to the molecular dynamics simulation (5 ns), and the results on the occurrence of average anchor points showed a stable complex between the benzoxazinone derivative and the receptor. The complex could be used as a good starting point for further analysis with site-directed mutagenesis, or further computational research. Figure The location of the ligands (complex B – blue; complex E – red; and complex F – green) in the transmembrane regions (TM1 – red; TM2 – blue; TM3 – yellow; TM4 – purple; TM5 – orange; TM6 – cyan; TM7 – pink) of the hOTR. For clarity, the EC and IC loops are not shown Electronic Supplementary Material Supplementary material is available at     

The activity of the RGA5 sensor NLR from rice requires binding of its integrated HMA domain to effectors but not HMA domain self-interaction

The rice nucleotide-binding (NB) and leucine-rich repeat (LRR) domain immune receptors (NLRs) RGA4 and RGA5 form a helper NLR/sensor NLR (hNLR/sNLR) pair that specifically recognizes the effectors AVR-Pia and AVR1-CO39 from the blast fungus Magnaporthe oryzae. While RGA4 contains only canonical NLR domains, RGA5 has an additional unconventional heavy metal-associated (HMA) domain integrated after its LRR domain. This RGA5_HMA domain binds the effectors and is crucial for their recognition. Investigation of the three-dimensional structure of the AVR1-CO39/RGA5_HMA complex by X-ray crystallography identified a candidate surface for effector binding in the HMA domain and showed that the HMA domain self-interacts in the absence of effector through the same surface. Here, we investigated the relevance of this HMA homodimerization for RGA5 function and the role of the RGA5_HMA effector-binding and self-interaction surface in effector recognition. By analysing structure-informed point mutations in the RGA5_HMA-binding surface in protein interaction studies and in Nicotiana benthamiana cell death assays, we found that HMA self-interaction does not contribute to RGA5 function. However, the effector-binding surface of RGA5_HMA identified by X-ray crystallography is crucial for both in vitro and in vivo effector binding as well as effector recognition. These results support the current hypothesis that noncanonical integrated domains of NLRs act primarily as effector traps and deepen our understanding of the sNLRs' function within NLR pairs.     

革兰氏阴性菌TonB依赖性受体的功能与结构研究进展*

为维持生长所需,革兰氏阴性菌需要从外界摄取多种营养物质。分子量小于600 Da的分子可以通过自由扩散的方式通过革兰氏阴性菌的外膜,而大分子物质则需要特殊的转运系统才能将其转运至革兰氏阴性菌的胞内。革兰氏阴性菌对大分子营养物质的识别和转运主要由TonB依赖性受体负责完成。所有革兰氏阴性菌中均有TonB依赖性受体的存在,然而不同种类的革兰氏阴性菌拥有TonB依赖性受体的数量不同且功能各异。最近研究表明,TonB依赖性受体不仅参与了铁、血红素、锰、锌、镍、维生素、碳水化合物等多种营养物质的摄取,而且参与了蛋白酶的分泌。为对TonB依赖性受体提供更为深入和系统的理解,详细介绍了目前已知的TonB依赖性受体的功能及结构,以期为更进一步探知TonB依赖性受体未知功能提供可参考依据。     

工程化外泌体介导巨噬细胞清除肿瘤外泌体*

目的:通过膜表面修饰改造技术构建工程化外泌体(engineered exosomes,enExos),并以此介导巨噬细胞特异性清除膜表面富含表皮生长因子受体(epidermal growth factor receptor,EGFR)的肿瘤外泌体。方法:利用表面展示技术获得膜表面展示趋化因子(chemokine 8,CXCL8)的外泌体,同时在其磷脂双分子层上修饰EGFR核酸适配体制备工程化外泌体;纳米颗粒跟踪和纳米粒度电位分析enExos的尺寸、电位;CCK-8试剂盒检测细胞活力;透射电子显微镜观察enExos与高表达EGFR的肿瘤外泌体的特异性结合;荧光成像技术及流式细胞术分析探究enExos靶向趋化巨噬细胞吞噬高表达EGFR的肿瘤外泌体。结果:成功构建膜表面展示EGFR与CXCL8的工程化外泌体,enExos可以特异性识别并捕获高表达EGFR的肿瘤外泌体,同时利用其趋化因子CXCL8特异性靶向巨噬细胞膜表面趋化因子受体CXCR1/CXCR2,刺激巨噬细胞对肿瘤外泌体的捕获及清除。结论:工程化外泌体促进了特定肿瘤外泌体的清除,为后续深入研究工程化外泌体抑制癌症转移的作用奠定基础,并期望为癌症转移治疗提供新的研究方向。     

High-fat diet during sexual maturation induces hyperplastic differentiation of rat prostate and higher expression of AR45 isoform and ERα

We examined the consequences of high-fat diet (HFD) on prostate histophysiology in two periods along sexual maturation of rats and the impact on the gland in adulthood. After weaning, male Wistar rats were fed a balanced diet (4 % fat-C3, C6, C9) or a HFD (20 % fat- HF3, HF6, HF9) for 3, 6 or 9 weeks. Fat deposit weights, blood glucose and levels of serum testosterone and estrogen were measured. Prostate was evaluated for histology, proliferative and apoptotic cell index, and for the expression of androgen (AR), estrogen receptors type α (ERα) and aromatase. HFD did not affect estrogen levels and elevated serum testosterone only in HF9. HFD reduced prostate weight in HF6 and increased it in adulthood (HF9) but relative prostate weight was unchanged among groups. Cell proliferation, height and density were higher in epithelium of all HFD-groups, compared to controls, featuring the epithelial hyperplasia. Epithelial apoptosis was lower in HF9. HF3 and HF9 exhibited higher expressions of ERα, indicating that HFD triggers a new activation of ERα expression in the acinar epithelium. The content of prostatic aromatase was also elevated in HF9. Increased numbers of AR-positive cells were observed in all HFD groups, and western blotting analysis showed an increase in the truncated form of 45 kDa (AR45) and a reduction in the expression of 110 kDa-AR for HF3 and HF9. In conclusion, excessive dietary fats during sexual maturation of rats led to developmental programming of the prostate, inducing a hyperplastic status with perturbations in AR isoforms expression and reactivation of ERα in adulthood, whose implications for posterior prostatic health could be detrimental.     

纳布啡鞘内注射对糖尿病神经痛模型大鼠行为能力及背根神经节TRPV1表达的影响

摘要 目的：探讨纳布啡鞘内注射对糖尿病神经痛（diabetic neuropathic pain, DNP）模型大鼠行为能力及背根神经节瞬时受体电位V1（transient receptor potential V1,TRPV1）表达的影响。方法：将糖尿病神经痛模型大鼠（n=48）随机平方为三组-模型组、纳布啡1组与纳布啡2组,每组16只。纳布啡1组与纳布啡2组分别给予纳布啡鞘内注射0.5 μg/10 μL与1.0 μg/10 μL,模型组给予注射等剂量的0.9 %氯化钠溶液,每天1次。分别于治疗第7 d、第14 d,采用血糖仪测定与记录空腹血糖（fasting blood glucose,FBG）水平并进行机械痛阈检测;治疗第14 d、第28 d,采用酶联免疫法检测血清IL-6与TNF-α含量,采用免疫印迹法检测背根神经节TRPV1蛋白的相对表达。结果：模型组、纳布啡1组与纳布啡2组治疗第7 d、第14 d的空腹血糖水平都高于20.00 mmol/L,组内与组间对比差异不具有统计学意义（P>0.05）。纳布啡1组与纳布啡2组治疗第7 d、第14 d的机械痛阈高于模型组（P<0.05）,也高于治疗前（P<0.05）,纳布啡2组与纳布啡1组差异具有统计学意义（P<0.05）。纳布啡1组与纳布啡2组治疗第14 d、第28 d的血清白细胞介素（Interleukin,IL）-6与肿瘤坏死因子（Tumor necrosis factor,TNF）-α含量明显低于模型组（P<0.05）,纳布啡2组明显低于纳布啡1组（P<0.05）。纳布啡1组与纳布啡2组治疗第14 d、第28 d的背根神经节TRPV1相对表达水平明显低于模型组（P<0.05）,纳布啡2组明显低于纳布啡1组（P<0.05）。结论：纳布啡鞘内注射在糖尿病神经痛模型大鼠的应用能改善行为能力,抑制背根神经节TRPV1的表达,还可抑制血清IL-6与TNF-α的释放,从而发挥镇痛治疗效用。     

三角梅苞叶色彩参数和色素含量分析及蓝色转基因受体的筛选北大核心CSCD

三角梅作为重要的观赏植物颜色繁多,但缺乏稀有的蓝色。为筛选适合的蓝色转基因受体,明确不同品种三角梅苞片吸收利用DHM(二氢杨梅素)合成甜菜色素途径竞争产物(类黄酮色素)的潜在能力,该研究对红色、白色、黄色和紫色4大花色6个品种的三角梅苞片进行离体诱导培养,测定诱导培养后苞片色彩参数及色素含量变化,并进行相关性分析。结果显示:(1)三角梅苞片红绿色相值(a*)是决定苞片呈色的主要色彩参数,其色彩主要由甜菜色素和黄酮类色素决定,并以甜菜红素的影响最大。(2)除白色品种三角梅苞片中黄酮类色素含量大于甜菜色素含量外,其余品种苞片发育中甜菜色素含量均呈上升趋势,黄酮类色素呈下降趋势。(3)甜菜色苷含量与苞片a*值呈显著正相关关系,同时与苞片黄蓝色相值(b*)呈显著负相关关系;总黄酮含量与苞片b*值呈显著正相关关系,与苞片a*值呈极显著负相关关系。(4)经DHM体外诱导培养后,苞片总黄酮含量及占比在4个品种三角梅(‘新加坡大白’、‘宝老橙’、‘中国丽人’、‘黄蝶’)中明显升高,但各品种苞片总甜菜色素含量及占比均下降,并以‘新加坡大白’苞片中总黄酮含量上升幅度最大(65.77%),含量占比变化(增加26.91%)也为6个品种中最大。(5)灰色关联度综合分析显示,白色品种‘新加坡大白’与灰色关联度分析拟定的参考品种关联度最高(0.7444),表明三角梅品种中‘新加坡大白’可考虑作为蓝色转基因三角梅的受体品种。     

植物M/MLD类受体蛋白激酶及其生物学功能的研究进展北大核心CSCD

植物受体蛋白激酶通过与胞外信号结合感知和接收外部信号传递,在植物各个生理过程及生物代谢中发挥着重大的作用。其中M/MLD类受体蛋白激酶是一类植物特有的具有Malectin-like结构域的受体蛋白激酶。研究表明,M/MLD-RLKs亚家族参与植物发育过程及生物/非生物胁迫调控。该研究对近年来国内外有关植物M/MLD-RLKs的发现、结构特点以及生物学功能等方面的研究进展进行综述,并重点阐述其在调控植物根系、叶片、花发育及响应多种胁迫过程中的作用,为深入研究M/MLD-RLKs在植物生长发育过程中的生理功能提供参考。     

猪环状RNA IGF1R促进脂肪细胞分化

环状RNA(circular RNA, circRNA)作为竞争性内源RNA(competitive endogenous RNA, ceRNA)在细胞分化调控中发挥着重要作用。本研究旨在对猪环状RNA IGF1R(circular RNA insulin-like growth factor 1 receptor, circIGF1R)进行鉴定及分析,探明其表达规律,构建猪circIGF1R相关的ceRNA调控网络,并探究其异位表达对小鼠间充质干细胞(C3H10T1/2)成脂分化的调控作用。通过正反向引物PCR、Sanger测序、RNase R酶消化检测和qRT-PCR验证circIGF1R是胰岛素样生长因子1受体(insulin-like growth factor 1 receptor, IGF1R)第二外显子形成的circRNA,它在猪各组织中均有表达,且其表达量在脂肪组织中随日龄增加呈上升趋势;使用miRDB、TargetScan和miRWalk在线软件预测circIGF1R靶基因,运用RNAhybrid软件进行结合位点预测,使用DAVID生物信息功能分析软件对候选靶基因进行GO和KEGG富集分析,运用Cytoscape软件构建ceRNA网络,基于基因表达相关性和预测的靶标关系,绘制了GO和KEGG富集分析及构建了ceRNA网络;双荧光素酶报告基因分析证明circIGF1R及FABP4可与ssc (Sus scrofa chromosome) -miR-133a-5p结合;成功构建circIGF1R过表达载体,在间充质干细胞C3H10T1/2中异位表达,过表达circIGF1R后关键成脂调控因子CEBPα、CEBPβ、FABP4和PPARγ极显著升高(P＜0.01),脂滴数量显著增加。本研究结果证明,circIGF1R在猪脂肪组织中存在,并且可能通过ceRNA机制正调控C3H10T1/2细胞成脂分化,为进一步研究circIGF1R调控猪前体肌内脂肪细胞成脂分化奠定理论基础。     

PM2.5通过激活NLRP3/Caspse-1通路诱导大鼠子宫炎症反应

颗粒物(PM)对呼吸系统、心血管系统、神经系统和免疫系统均有损害,但目前关于吸入颗粒物对生殖损伤的研究较少。本研究旨在探讨细颗粒物(PM_2.5)短期暴露对大鼠子宫炎症损伤及其作用机制。PM_2.5暴露30 d后,高剂量组大鼠的子宫脏器系数、内膜上皮细胞厚度和腺上皮高度均明显高于对照组(P<0.05),抑制剂MCC950则能明显降低PM_2.5对子宫的影响。子宫组织免疫荧光双染色结果显示,PM_2.5暴露组子宫内CD45白细胞和CD11b巨噬细胞均明显增加(P<0.05)。Elisa法检测子宫组织和血清中白介素1β(IL-1β)和转化生长因子-β1(TGF-β1),暴露组子宫组织和血清中IL-1β和TGF-β1含量明显升高(P<0.05)。Western印迹法检测结果显示,PM_2.5上调核苷酸结合低聚体结构域样受体3 (NLRP3)、凋亡相关斑点样蛋白质(ASC)、pro-IL-1β、pro-Caspase-1和半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)的蛋白质表达量(P<0.05)。与高剂量组相比,NLRP3抑制剂MCC950能明显降低NLRP3/Caspase-1通路中关键蛋白质表达水平(P<0.05)。综上,PM_2.5通过激活NLRP3/ Caspase-1信号,诱导大鼠子宫炎症反应,为PM_2.5生殖毒性预防和治疗提供理论基础。