Structural Insights into the Epimerization of β-1,4-Linked Oligosaccharides Catalyzed by Cellobiose 2-Epimerase,the Sole Enzyme Epimerizing Non-anomeric Hydroxyl Groups of Unmodified Sugars

Cellobiose 2-epimerase (CE) reversibly converts d-glucose residues into d-mannose residues at the reducing end of unmodified β1,4-linked oligosaccharides, including β-1,4-mannobiose, cellobiose, and lactose. CE is responsible for conversion of β1,4-mannobiose to 4-O-β-d-mannosyl-d-glucose in mannan metabolism. However, the detailed catalytic mechanism of CE is unclear due to the lack of structural data in complex with ligands. We determined the crystal structures of halothermophile Rhodothermus marinus CE (RmCE) in complex with substrates/products or intermediate analogs, and its apo form. The structures in complex with the substrates/products indicated that the residues in the β5-β6 loop as well as those in the inner six helices form the catalytic site. Trp-322 and Trp-385 interact with reducing and non-reducing end parts of these ligands, respectively, by stacking interactions. The architecture of the catalytic site also provided insights into the mechanism of reversible epimerization. His-259 abstracts the H2 proton of the d-mannose residue at the reducing end, and consistently forms the cis-enediol intermediate by facilitated depolarization of the 2-OH group mediated by hydrogen bonding interaction with His-200. His-390 subsequently donates the proton to the C2 atom of the intermediate to form a d-glucose residue. The reverse reaction is mediated by these three histidines with the inverse roles of acid/base catalysts. The conformation of cellobiitol demonstrated that the deprotonation/reprotonation step is coupled with rotation of the C2-C3 bond of the open form of the ligand. Moreover, it is postulated that His-390 is closely related to ring opening/closure by transferring a proton between the O5 and O1 atoms of the ligand.     

草原群落退化演替过程中微斑块土壤碳氮的空间异质动态

微斑块变化是草原退化过程中的活跃成分。分析了呼伦贝尔克氏针茅草原逆行演替过程中微斑块土壤全碳、全氮和碱解氮含量的空间异质性,提出了"养分聚集效应"的概念。研究结果表明:随着群落退化演替的加剧,土壤全碳、全氮和碱解氮的含量均表现为演替前期演替后期演替中期(P0.05)。从土壤全碳、全氮和碱解氮的变异系数和变异函数综合分析来看,10 cm×10 cm微尺度上,草原退化演替过程中土壤全碳、全氮和碱解氮的空间异质性具有明显的不一致性;全碳的空间异质性表现为演替中期演替前期演替后期,全氮表现为演替后期演替前期演替中期,碱解氮表现为演替中期演替后期演替前期。草原退化过程中土壤养分在微斑块上的富积和迁移表现出尺度依赖性和变异性。     

我国三种蝼蛄的雄性生殖器鉴别

本文记述 3种蝼蛄 (单刺蝼蛄GryllotalpaunispinaSaussure,东方蝼蛄G .orientalisBurmeister及河南蝼蛄G .henanaCaietNiu)的雄生殖器     

Adaptive T cell immunotherapy in cancer

Impaired tumor-specific effector T cells contribute to tumor progression and unfavorable clinical outcomes. As a compensatory T cell-dependent cancer immunoediting strategy, adoptive T cell therapy(ACT) has achieved encouraging therapeutic results,and this strategy is now on the center stage of cancer treatment and research. ACT involves the ex vivo stimulation and expansion of tumor-infiltrating lymphocytes(TILs) with inherent tumor reactivity or T cells that have been genetically modified to express the cognate chimeric antigen receptor or T cell receptor(CAR/TCR), followed by the passive transfer of these cells into a lymphodepleted host. Primed T cells must provide highly efficient and long-lasting immune defense against transformed cells during ACT. Anin-depth understanding of the basic mechanisms of these living drugs can help us improve upon current strategies and design better next-generation T cell-based immunotherapies. From this perspective, we provide an overview of current developments in different ACT strategies, with a focus on frontier clinical trials that offer a proof of principle. Meanwhile, insights into the determinants of ACT are discussed, which will lead to more rational, potent and widespread applications in the future.     

EL转染试剂转染食管鳞癌细胞的条件优化

该文探讨了关于EL转染试剂转染Hsa-miR-6743质粒至食管鳞癌细胞转染效果的影响因素.以食管鳞癌细胞株Eca-109、TE-1和Eca-9706为研究对象,GFP标记的Hsa-miR-6743为报告基因,通过倒置荧光显微镜检测荧光信号优化转染试剂和质粒比值.结果表明,食管鳞癌细胞的种类影响EL转染试剂的转染效果,...     

Crumbs proteins stabilize the cone mosaics of photoreceptors and improve vision in zebrafish

Although the unique organization of vertebrate cone mosaics was first described long ago,both their underlying molecular basis and physiological significance are largely unknown.Here,we demonstrate that Crumbs proteins,the key regulators of epithelial apical polarity,establish the planar cellular polarity of photoreceptors in zebrafish.Via heterophilic Crb2a-Crb2b interactions,the apicobasal polarity protein Crb2b restricts the asymmetric planar distribution of Crb2a in photoreceptors.The planar polarized Crumbs proteins thus balance intercellular adhesions and tension between photoreceptors,thereby stabilizing the geometric organization of cone mosaics.Notably,loss of Crb2b in zebrafish induces a nearsightedness-like phenotype in zebrafish accompanied by an elongated eye axis and impairs zebrafish visual perception for predation.These data reveal a detailed mechanism for cone mosaic homeostasis via previously undiscovered apical-planar polarity coordination and propose a pathogenic mechanism for nearsightedness.     

LncRNA RP11-89 facilitates tumorigenesis and ferroptosis resistance through PROM2-activated iron export by sponging miR-129-5p in bladder cancer

Long non-coding RNAs (lncRNAs) act as important regulators of tumorigenesis and development in bladder cancer. However, the underlying molecular mechanisms remain elusive. We previously identified a novel lncRNA signature related to immunity and progression in bladder cancer. Here we further explored the function of RP11-89, a lncRNA discovered in the previous signature. Loss- and gain-of function experiments were performed using CCK-8 assay, flow cytometry, Transwell assays, scratch tests and subcutaneous nude mouse models. High-throughput RNA sequencing was conducted to identify dysregulated genes in bladder cancer cells with RP11-89 knockdown or overexpression. Regulation of RP11-89 on miR-129-5p and PROM2 was explored through luciferase reporter assay, RIP assay and RNA pull-down assay. RP11-89 promoted cell proliferation, migration and tumorigenesis and inhibited cell cycle arrest via the miR-129-5p/PROM2 axis. We found that RP11-89 “sponges” miR-129-5p and upregulates PROM2. Elevated PROM2 in cells was associated with attenuated ferroptosis through iron export, formation of multivesicular bodies and less mitochondrial abnormalities. We demonstrated that RP11-89 is a novel tumorigenic regulator that inhibits ferroptosis via PROM2-activated iron export. RP11-89 may serve as a potential biomarker for targeted therapy in bladder cancer.Subject terms: Tumour biomarkers, Tumour biomarkers     

江西早稻高温热害等级动态判识及时空变化特征

构建考虑高温天气过程(包括发生时间、持续日数)的水稻高温热害指标,可以实现对水稻高温热害等级的动态判识,对精准监测、预警与评估水稻高温热害意义重大。以江西早稻为对象,利用气象资料、早稻高温热害灾情史料和生育期资料,反演历史早稻高温热害,采用K-S分布拟合检验和置信区间方法,构建基于高温天气过程的早稻高温热害动态指标,并采用预留的独立的早稻高温热害样本进行检验验证。在此基础上,计算江西各站点早稻高温热害指数(M),并分析水稻热害。结果表明: 高温天气过程起始时间、持续日数是影响早稻高温热害发生程度的关键因子,其中,起始时间的影响大于持续日数。3～5 d早稻轻、中、重度高温热害的起始时间阈值分别为抽穗后第10～12、5～9、2～4天;6～8 d早稻轻、中、重度高温热害的起始时间阈值为抽穗后第11～18、8～10和1～7天;>8 d早稻轻、中、重度高温热害的起始时间阈值为抽穗后第12～18、8～11和0～7天。指标验证完全一致的吻合率为73.7%,完全一致及相差1级的吻合率为89.5%。1981—2015年,M的线性倾向率为0.04·a^-1,1999年左右发生由低到高突变;M高值区域主要位于江西中部和东北部,M>0.18;江西中部、东北部和南部地区M值呈显著增加趋势,线性倾向率均大于0.04·a^-1。总体来说,本文构建的指标实现了基于高温天气过程的早稻高温热害动态判识,江西中部和东北部是早稻高温热害的高风险区域。