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.     

Pyrophosphate contamination in crystalline phosphocreatine: a source of error in kinetic studies using coupled enzymes.

Commercial crystalline phosphocreatine generally contains a contaminant, probably pyrophosphate, that can inhibit nucleoside triphosphatases and may therefore cause errors in steady-state kinetic studies of creatine kinase-coupled r reactions. Ion-exchange chromatography reveals the presence of pyrophosphate in some batches of phosphocreatine sufficient to account for the observed inhibition. Chromatographically purified phosphocreatine shows no inhibitory effect.     

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。     

DAZL regulates proliferation of human primordial germ cells by direct binding to precursor miRNAs and enhances DICER processing activity

Understanding the molecular and cellular mechanisms of human primordial germ cells (hPGCs) is essential in studying infertility and germ cell tumorigenesis. Many RNA-binding proteins (RBPs) and non-coding RNAs are specifically expressed and functional during hPGC developments. However, the roles and regulatory mechanisms of these RBPs and non-coding RNAs, such as microRNAs (miRNAs), in hPGCs remain elusive. In this study, we reported a new regulatory function of DAZL, a germ cell-specific RBP, in miRNA biogenesis and cell proliferation. First, DAZL co-localized with miRNA let-7a in human PGCs and up-regulated the levels of >100 mature miRNAs, including eight out of nine let-7 family, miR21, miR22, miR125, miR10 and miR199. Purified DAZL directly bound to the loops of precursor miRNAs with sequence specificity of GUU. The binding of DAZL to the precursor miRNA increased the maturation of miRNA by enhancing the cleavage activity of DICER. Furthermore, cell proliferation assay and cell cycle analysis confirmed that DAZL inhibited the proliferation of in vitro PGCs by promoting the maturation of these miRNAs. Evidently, the mature miRNAs up-regulated by DAZL silenced cell proliferation regulators including TRIM71. Moreover, DAZL inhibited germline tumor cell proliferation and teratoma formation. These results demonstrate that DAZL regulates hPGC proliferation by enhancing miRNA processing.     

Effects of highly selective sensory/motor nerve injury on bone metabolism and bone remodeling in rats

Objectives:This work aimed to investigate the mechanism of selective sensory/motor nerve injury in affecting bone metabolism and remodeling.Methods:The selective sensory/motor nerve injury rat model was constructed through posterior rhizotomy (PRG), anterior rhizotomy (ARG), or anterior combined with posterior rhizotomy (APRG) at the L_4-6 sensory/motor nerves on the right side of rats. Sham-operated (SOG) rats served as control. At 8 weeks after surgery, the sciatic nerves, spinal cord segments L₅ and tibial tissues were collected for analysis.Results:the integrity of trabecular bone was damaged, the number of trabecular bone was decreased and the number of osteoclasts were increased in ARG group. ARG activated NF-κβ and PPAR-γ pathways, and inhibited Wnt/β-catenin pathway. ARG group exhibited high turnover bone metabolism. In PRG group, the trabecular bone morphology became thinner, and the number of osteoclasts was increased. NF-κβ pathway was activated and OPG/RANKL ratio was decreased in PRG group. The activated osteoclasts, reduced osteoblasts activity and lower turnover bone metabolism were observed in PRG group. Additionally, the bone metabolism in APRG group was similar to ARG group.Conclusion:The posterior rhizotomy and anterior rhizotomy induced the different degree of osteoporosis in rats, which may attribute to regulate Wnt/β-catenin, NF-κβ and PPAR-γ signalling pathways.