上海地区胃癌病人运铁蛋白(Tf)、组特异性成分(Gc)、α_1-抗胰蛋白酶(α_1-AT)亚型分布的研究

用薄层聚丙烯酰胺凝胶等电聚焦电泳分析了上海地区202例汉族胃癌病人及202例正常对照组的运铁聋白(Tf)和α_1-抗胰蛋白酶(α_1-AT)亚型的分布,发现胃癌组Tfc_1c_1纯合子频率(0.3713)和Tfc_1基因频率(0.5718)显著低于对照组(分别为0.5149和0.6782),均为p<0.01,胃癌组Tfc_2c_2纯合子频率(0.2228)和Tfc_2基因频率(0.4019)显著高于对照组(分别为0.1436,p<0.05和0.2970,p<0.01),胃癌组和对照组的α_1-抗胰蛋白酶亚型分布无显著差异。用薄层聚丙烯酰胺等电聚焦结合免疫固定分析了上海地区200例汉族胃癌病人和200例正常对照组的组特异性成分(Gc)亚型分布,发现胃癌组Go1F表型频率(0.22)和Gc1F基因频率(0.4375)均显著高于正常对照组(分别为0.14和0.3600,均为p<0.05)。     

Detection of covalent enzyme-substrate complexes of nitrilase by ion-spray mass spectroscopy

Nitrilase from Rhodococcus ATCC 39484 was found to consist of two species of Mr 40,258 +/- 2 and 40,388 +/- 2 Da. When the enzyme was incubated with nitrile substrates and the reaction quenched with acid, higher Mr species were observed. The mass differences were consistent with addition of a substrate molecule to each species. These results represent the first reported demonstration that this, or any other nitrilase forms a covalent intermediate with its substrates. The observation that the intermediate, suggested to be either a thioimidate or an acylenzyme, can be trapped by acidification indicates that the rate of breakdown of the intermediate is rate-limiting.     

Vibrational fluctuations of hydrogen bonds in a DNA double helix with nonuniform base pairs.

The Green's function technique is applied to a study of breathing modes in a DNA double helix which contains a region of different base pairs from the rest of the double helix. The calculation is performed on a G-C helix in the B conformation with four consecutive base pairs replaced by A-T. The average stretch in hydrogen bonds is found amplified around the A-T base pair region compared with that of poly(dG)-poly(dC). This is likely related to the A-T regions lower stability against hydrogen bond melting. The A-T region may be considered to be the initiation site for melting in such a helix.     

A method for separating bound versus unbound label during radioiodination

An inexpensive, highly effective, and safe method for the removal of bound versus unbound label during radiolabeling of proteins is described. The technique employs the use of membrane ultrafiltration technology and returns in one step a highly reproducible product of quality superior to that attained by gel chromatography. Advantages of this technique are a reduction in the quantity of liquid and solid radioactive waste and a significant limitation of potentially harmful manipulation and exposure times.     

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.     

Sound localization behavior of the green treefrog (Hyla cinerea) and the barking treefrog (H. gratiosa)

Summary Female treefrogs (Hyla cinerea andH. gratiosa) can accurately localize a sound source (playback of male mating calls) if both ears are intact. When the sensitivity of one eardrum is attenuated, by coating it with a thin layer of silicone grease, females no longer can locate the sound source. This study demonstrates that female anurans rely on interaural cues for localization of a calling male. The neural basis for an anuran's sound localization ability presumably involves binaural convergence on single cells in the central auditory nervous system.This work was supported by research grants from the National Science Foundation and the U.S. Public Health Service. The assistance of Anne J.M. Moffat in measuring the directional characteristics of the loudspeaker is gratefully appreciated.     

用EDTA法研究镰刀菌胞内、胞外Pb2+的分布特征

【目的】建立一种方便、快捷、相对准确、能够定量地测定镰刀菌细胞内、外Pb2+分布的技术手段。【方法】用EDTA溶液浸泡镰刀菌细胞,使其胞外(表面)Pb2+被螯合、洗脱并测定,之后将被浸泡、清洗过的细胞消解、测铅。【结果】EDTA可以将镰刀菌表面的Pb2+螯合,且在99 min内不损伤镰刀菌细胞;以EDTA为反应介质和滴定剂,XO为指示剂,测定镰刀菌胞内、胞外Pb2+分布是可行的。依据此实验方法,测定了镰刀菌在Pb2+浓度为500 mg/L的培养基中的生长曲线、培养基中Pb2+浓度和细胞内、外Pb2+的含量。【结论】镰刀菌固定Pb2+的过程是先将Pb2+吸附在菌体胞外,之后转运至细胞内部,菌体胞外Pb2+的容纳量是有限的,每克菌体胞外Pb2+饱和吸附量约1.37 mg,通过计算可得,每克菌体用于吸附Pb2+的胞外活性位点约3.97×1018个。     

资源型城市土地利用转型及其对生境质量的影响——以乌海市为例

资源型城市前期发展导致了生境丧失或退化,实现高质量转型需要深入理解城市转型与生态环境质量之间的关系,土地利用转型特征及其对生境质量的影响规律研究为此提供依据与支撑。以资源型城市--乌海市为研究区,通过地学信息图谱和InVEST模型探究2005-2018年乌海市土地利用转型特征、生境质量时空变化及土地利用转型对生境质量的影响。结果表明：（1）2005-2018年乌海市土地利用变化趋势发生改变,土地利用转型明显,土地利用变化图谱单元数量逐渐增加78.14%,分布范围逐渐广泛。主要表现为草地与建设用地、采矿用地之间的相互转化,第一阶段（2005-2015年）草地大面积减少,建设用地和采矿用地大面积增加,第二阶段（2015-2018年）趋势相反。（2）乌海市生境质量变化呈现先强退化后弱提升趋势。2005-2015年乌海市18.75%的区域生境质量退化,提升面积较小;2015-2018年生境质量提升面积略大于退化面积。（3）2005-2015年草地向采矿用地、建设用地转化是区域生境质量降低的主要原因,2015-2018年区域生境质量提升的主导因素是采矿用地和建设用地向草地转化。研究结果揭示了资源型城市土地利用转型过程中生境质量的响应规律,可为资源型城市土地利用转型决策提供参考。     

IGF2BP2 serves as a core m6A regulator in head and neck squamous cell carcinoma

Methylation of N6 adenosine (m6A) plays a crucial role in the development and progression of cancers. Its modification is regulated by three types of m6A-related regulators (methyltransferases (writers), demethylases (erasers), and RNA-binding proteins (readers)). Till now, the functions and roles of these regulators in head and neck squamous cell carcinoma (HNSC) remain largely unexplored. Therefore, we utilized the open HNSC dataset in The Cancer Genome Atlas (TCGA), four different cell lines, and our HNSC patient samples (n=40) to explore the clinical significance of 19 m6A regulators, and selected the most significant prognosis-related regulator. Authentic analyses based on online websites were also used in the study (Oncomine, UALCAN, Kaplan–Meier plotter, Human Protein Atlas (HPA), cBioPortal, LinkedOmics, String, etc.). From the results, general overexpression of m6A regulators was observed in pan-cancer, especially in HNSC. IGF2BP2 was recognized as the hub m6A regulator, which was an independent, unfavorable prognostic factor in HNSC. Its mRNA and protein expression in HNSC were significantly up-regulated. Gene mutation types of IGF2BP2 in HNSC (32%) were mainly mRNA High or Amplification, which represented the high expression of IGF2BP2. And these mutations were associated with a poor prognosis. In functional analysis, IGF2BP2 was negatively correlated to tumor immune infiltration in HNSC. Finally, HMGA2 might interact with the IGF2BP2 in HNSC. In conclusion, IGF2BP2 serves as a core m6A regulator among all regulators in HNSC, which has a high expression and predicts the poor prognosis of HNSC patients independently. IGF2BP2 might bring a new direction for HNSC treatment in the future.     

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.