The conformation of single-stranded nucleic acids tDNA versus tRNA

Conformational analyses using the single-strand-specific nuclease from mung bean and restriction endonucleases have been performed on a series of DNA fragments related to the sequence of the yeast initiator tRNA(Met). Mung bean nuclease cleaves DNA fragments exclusively in some, but not all, single-stranded regions as predicted by RNA secondary structural rules. Comparison of cleavage patterns of yeast initiator tRNA(Met), tDNA(Met) (a DNA oligomer having the sequence of tRNA(Met] and the anti-tDNA(Met) (the complement of tDNA(Met] suggests that the conformation of the three molecules is very similar. Furthermore, both tDNA and anti-tDNA are cleaved by HhaI and CfoI restriction endonucleases at two GCG/C sites which would be in double-stranded regions (the acceptor and dihydrouridine stem), if the two molecules adopt the tRNA cloverleaf structure. On the other hand, minor cleavage products show that the core region, i.e. the extra loop area, is slightly more exposed in tDNA and in anti-tDNA than in tRNA. Therefore, we submit that the global conformation of nucleic acids is primarily dictated by the interaction of purine and pyrimidine bases with atoms and functional groups common to both RNA and DNA. In this view the 2'-hydroxyl group, in tRNA at least, is an auxiliary structural feature whose role is limited to fostering local interactions, which increase the stability of a given conformation.     

Cloning and sequencing of a human pancreatic tumor mucin cDNA

A monospecific polyclonal antiserum against deglycosylated human pancreatic tumor mucin was used to select human pancreatic mucin cDNA clones from a lambda gt11 cDNA expression library developed from a human pancreatic tumor cell line. The full-length 4.4-kilobase mucin cDNA sequence included a 72-base pair 5'-untranslated region and a 307-base pair 3'-untranslated region. The predicted amino acid sequence for this cDNA revealed a protein of 122,071 daltons containing 1,255 amino acid residues of which greater than 60% were serine, threonine, proline, alanine, and glycine. Approximately two-thirds of the protein sequence consisted of identical 20-amino acid tandem repeats which were flanked by degenerate tandem repeats and nontandem repeat sequences on both the amino-terminal and carboxyl-terminal ends. The amino acid sequence also contained five putative N-linked glycosylation sites, a putative signal sequence and transmembrane domain, and numerous serine and threonine residues (potential O-linked glycosylation sites) outside and within the tandem repeat position. The cDNA and deduced amino acid sequence of the pancreatic mucin sequence was over 99% homologous with a mucin cDNA sequence derived from breast tumor mucin, even though the native forms of these molecules are quite distinct in size and degree of glycosylation.     

CD36 aggravates podocyte injury by activating NLRP3 inflammasome and inhibiting autophagy in lupus nephritis

A major cause of proteinuria in lupus nephritis (LN) is podocyte injury, and determining potential therapeutic targets to prevent podocyte injury is important from a clinical perspective in the treatment of LN. CD36 is involved in podocyte injury in several glomerulopathies and was reported to be a vital candidate gene in LN. Here, we determined the role of CD36 in the podocyte injury of LN and the underlying mechanisms. We observed that CD36 and NLRP3 (NLR family pyrin domain containing 3) were upregulated in the podocytes of lupus nephritis patients and MRL/lpr mice with renal impairment. In vitro, CD36, NLRP3 inflammasome, and autophagy were elevated accompanied with increased podocyte injury stimulated by IgG extracted from lupus nephritis patients compared that from healthy donors. Knocking out CD36 with the CRISPR/cas9 system decreased the NLRP3 inflammasome levels, increased the autophagy levels and alleviated podocyte injury. By enhancing autophagy, NLRP3 inflammasome was decreased and podocyte injury was alleviated. These results demonstrated that, in lupus nephritis, CD36 promoted podocyte injury by activating NLRP3 inflammasome and inhibiting autophagy by enhancing which could decrease NLRP3 inflammasome and alleviate podocyte injury.Subject terms: Mechanisms of disease, Inflammasome, Lupus nephritis, Autophagy     

迁地麋鹿对野鸭湖植被的保护性生物控制

利用食草动物来管理自然保护地的植被平衡具有很大的应用潜力,一方面可提升动物的生态价值,另一方面通过控制取食规模,改变植被的生物多样性,达到对自然保护地生态平衡管理的目的。基于此于2021年6月5日引入4头麋鹿(2雄2雌),对野鸭湖自然保护区的“芦苇优势群落”采取保护性的生物控制研究,从项目的实施来看：1)单纯收割不能控制芦苇的生长扩张;对芦苇区系植物多样性的影响有限,未改变芦苇区系结构;2)麋鹿引入该区域后,通过取食、游泳、躺卧和踩踏等活动有效控制了芦苇和香蒲的过度扩张;1年后芦苇和香蒲面积下降了21.96%,为三棱水葱、水蓼等提供了生长空间,逐渐形成了仍以芦苇和香蒲为主且更多样的湿地环境;3)增加滩涂和开阔水面等景观,使多样性指数进一步提升,未改变周边区系湿地生态结构;4)麋鹿迁入可降低野鸭湖“脆弱物种”芦苇区系的丰富度,由引入前的(r=3.67)下降到引入后的(r=1.97);麋鹿迁入提升了野鸭湖植被区系物种多样性,芦苇区系的多样性指数由引进前的(r=0.90)上升到引进后的(r=2.11);麋鹿引入的第一年结果显示,整个引入区域的植被多样性指数由r=0.51上升到r=0.91。麋...     

高原湖泊流域国土空间生态修复优先区诊断及修复研究

高原湖泊流域是高原地区人类活动的重要载体,兼具高生态价值和高脆弱性的特点。随着高原湖泊流域城市化和工业化发展加速,湖泊面积萎缩,污染加剧,流域生态环境受损严重,引发了一系列生态环境问题,如水土流失、水污染、湿地退化、生境质量下降等。亟需开展生态修复以平衡经济发展与生态环境保护之间关系,而基于整体保护与系统治理思维诊断并修复生态修复优先区,是科学有序推进国土空间生态保护与修复的重要抓手。基于此,研究以高原湖泊流域典型代表滇池流域为例,利用人类足迹和景观生态风险模型定量评估生态系统所受负向干扰,以最小累积阻力模型和电路理论构建流域生态网络;提取生态网络受负向干扰较高的关键区域为生态修复优先区并提出针对性修复措施。研究表明：(1)滇池流域人类干扰和生态风险整体较高,人类干扰整体呈核心—边缘递减的圈层式分布,中高生态风险占据了绝大部分区域。人类交通网络大幅扩展了人类干扰和生态风险的强度和深度;(2)区域生态网络呈典型湖泊生态网络特点,38条生态廊道呈放射状或环状分布,连通湖区、山区两大生态空间内共23块生态源地,保障区域生态安全;(3)研究共提取生态源地修复优先区73.83km²     

黄土高原次生林演替对团聚体有机碳含量及化学稳定性的影响

次生林演替过程中土壤团聚体有机碳的积累机制和化学稳定性研究较少。为探明次生林演替对土壤团聚体有机碳含量及其化学组成稳定性的影响,选取黄土高原次生白桦林(演替初期),山杨辽东栎混交林(演替中期)和辽东栎林(演替后期)为研究对象,分析演替过程中不同粒径土壤团聚体有机碳含量变化特征。采用傅里叶红外光谱技术(FTRI)测定活性(AC)和非活性(IC)有机碳化学组成,以(IC/AC)作为有机碳化学组成稳定性指标,并分析其影响因素。结果表明:次生林演替过程中土壤团聚体有机碳含量表现出逐渐增加的趋势且各群落间差异显著(P<0.05),以演替后期的中等粒径团聚体为最高(37.63 g/kg)。土壤团聚体AC中多糖体有机碳含量最高(55.87%),而IC中芳香族有机碳含量最高(94.45%),演替过程中IC与AC总体变化趋势均呈现先降后增。IC/AC随着演替的进行呈先降低后升高的趋势,其中演替后期微团聚体有机碳化学组成稳定性最强达到了3.95。微团聚体含量(WM)与土壤全氮、全磷、全钾一起,显著促进了团聚体有机碳化学组成稳定性(P<0.05)。综上,次生林演替有利于促进土壤团聚体有机碳的积累以及有机碳化学稳定,其中微团聚体起到了关键性作用。     

粪产碱杆菌的分离鉴定及其生物转化作用

【背景】硫化氢(H2S)作为畜牧生产过程中释放的一种有毒有害气体,严重危害畜禽和人类的健康,因此降解硫化氢特别是生物氧化法转化硫化氢已成为当前研究热点。【目的】筛选高效硫氧化菌株并研究其生物转化作用。【方法】以长春市某养鸡场采集的新鲜粪便为材料,分离鉴定硫氧化菌株。采用单因素分析法优化其生长条件,研究生物转化效率,检测soxY、soxZ基因m RNA表达水平。【结果】获得一株高效硫氧化菌株JF9,经鉴定为粪产碱杆菌。最佳生长条件:底物浓度0.5 g/L,温度35°C,初始pH 7.0,在此条件下Na2S去除率达94%以上。菌株JF9存在soxY和soxZ基因,其转录水平在硫源诱导前后差异显著(P0.05)。【结论】分离得到的粪产碱杆菌具有良好的硫化物转化能力,脱硫过程中硫氧化基因高效表达。     

A toolbox for class I HDACs reveals isoform specific roles in gene regulation and protein acetylation

The class I histone deacetylases are essential regulators of cell fate decisions in health and disease. While pan- and class-specific HDAC inhibitors are available, these drugs do not allow a comprehensive understanding of individual HDAC function, or the therapeutic potential of isoform-specific targeting. To systematically compare the impact of individual catalytic functions of HDAC1, HDAC2 and HDAC3, we generated human HAP1 cell lines expressing catalytically inactive HDAC enzymes. Using this genetic toolbox we compare the effect of individual HDAC inhibition with the effects of class I specific inhibitors on cell viability, protein acetylation and gene expression. Individual inactivation of HDAC1 or HDAC2 has only mild effects on cell viability, while HDAC3 inactivation or loss results in DNA damage and apoptosis. Inactivation of HDAC1/HDAC2 led to increased acetylation of components of the COREST co-repressor complex, reduced deacetylase activity associated with this complex and derepression of neuronal genes. HDAC3 controls the acetylation of nuclear hormone receptor associated proteins and the expression of nuclear hormone receptor regulated genes. Acetylation of specific histone acetyltransferases and HDACs is sensitive to inactivation of HDAC1/HDAC2. Over a wide range of assays, we determined that in particular HDAC1 or HDAC2 catalytic inactivation mimics class I specific HDAC inhibitors. Importantly, we further demonstrate that catalytic inactivation of HDAC1 or HDAC2 sensitizes cells to specific cancer drugs. In summary, our systematic study revealed isoform-specific roles of HDAC1/2/3 catalytic functions. We suggest that targeted genetic inactivation of particular isoforms effectively mimics pharmacological HDAC inhibition allowing the identification of relevant HDACs as targets for therapeutic intervention.