棒状类细菌电击转化中多种条件对转化效率的影响

以质粒PXZl0145电击转化不同棒状类细菌菌株,研究了影响电击转化效率的诸个因素,在含4%甘氨酸的培养基中生长至对数前期的菌体最适用于电击转化,当以同源DNA进行电击转化时,1．μgDNA中转化效率最高可达到8×1O⁶转化子,但用异源DNA时,转化效率要比前者低10²～10³倍。     

Evidence forCis- andTrans-acting element coevolution of the 2-μm circle genome inSaccharomyces cerevisiae

Summary We compared the DNA sequence of the yeas 2-μm plasmidcis-actingSTB andtrans-actingREP1 partition loci of laboratory haploid and industrial amphiploid strains. Several industrial strains had a uniqueSTB sequence (type 1) sharing only 70% homology with laboratorySTB (type 2). Type 1 plasmids had a REP1 protein with 6–10% amino acid substitutions when compared to REP1 of type 2 plasmids. All 2-μm variants that shared a similarSTB consensus sequence exhibited a high degree ofREP1 nucleotide and amino acid sequence conservation. These observations suggest molecular coevolution oftrans-acting elements with cognate target DNA structure. Based on DNA sequencing and Southern hybridization analyses, we classified 2-μm variants into two main evolutionary lineages that differ atSTB as well asREP1 loci. The role of molecular coevolution in yeast intra- and interspecies plasmid evolution was discussed.     

Spatial and temporal patterns of intracellular calcium in colonic smooth muscle

Summary Intracellular calcium [Ca²⁺] _i measurements in cell suspension of gastrointestinal myocytes have suggested a single [Ca²⁺] _i transient followed by a steady-state increase as the characteristic [Ca²⁺] _i response of these cells. In the present study, we used digital video imaging techniques in freshly dispersed myocytes from the rabbit colon, to characterize the spatiotemporal pattern of the [Ca²⁺] _i signal in single cells. The distribution of [Ca²⁺] _i in resting and stimulated cells was nonhomogeneous, with gradients of high [Ca²⁺] _i present in the subplasmalemmal space and in one cell pole. [Ca²⁺] _i gradients within these regions were not constant but showed temporal changes in the form of [Ca²⁺] _i oscillations and spatial changes in the form of [Ca²⁺] _i waves. [Ca²⁺] _i oscillations in unstimulated cells (n = 60) were independent of extracellular [Ca²⁺] and had a mean frequency of 12.6 +1.1 oscillations per min. The baseline [Ca²⁺], was 171 ± 13 nm and the mean oscillation amplitude was 194 ± 12 nm. Generation of [Ca²⁺] _i waves was also independent of influx of extracellular Ca²⁺. [Ca²⁺] _i waves originated in one cell pole and were visualized as propagation mostly along the subplasmalemmal space or occasionally throughout the cytoplasm. The mean velocity was 23 +3 m per sec (n = 6). Increases of [Ca²⁺] _i induced by different agonists were encoded into changes of baseline [Ca²⁺] _i and the amplitude of oscillations, but not into their frequency. The observed spatiotemporal pattern of [Ca²⁺] _i regulation may be the underlying mechanism for slow wave generation and propagation in this tissue. These findings are consistent with a [Ca²⁺] _i regulation whereby cell regulators modulate the spatiotemporal pattern of intracellularly generated [Ca²⁺] _i oscillations.The authors thank Debbie Anderson for excellent technical assistance with the electron microscopy and Dr. M. Regoli for providing the NK-1 agonist [Sar⁹,Met(O₂)¹¹]-SP. This work was supported by National Institutes of Health Grants DK 40919 and DK 40675 and Veterans Administration Grant SMI.     

Distribution and behaviour of Acetes sibogae Hansen (Decapoda, Crustacea) in an estuary in relation to tidal and diel environmental changes

Quantitative samples of Acetes sibogae were collected at 2 hintervals for 48 h at three sites across the axis of a tidalestuary to examine their distribution within the water bodyover tidal and diel cycles, and to assess the role of behaviourin maintaining population distribution in estuarine/coastalwaters in relation to selected environmental factors. Watertemperature, salinity, tidal height and light intensity wereconcurrently measured. Distribution of the shrimp across theestuary was uniform and consistent between daylight or darkperiods, and among flood or ebb tides. Changes of A.sibogaeabundance were related to light and tidal cycles at each sitewith higher catches in dark periods and during flood tides.Acetes sibogae also exhibited both nocturnal and tidal verticalmovements in the water body, with greater numbers being onlyfound near-surface rather than near-bottom during flood tidesand at night. No significant differences in the distributionof size groups were found between any sampled levels of anysite. Acetes sibogae was highly aggregated in the water body.It is suggested that aggregating behaviour and tidal and nocturnalvertical movements act to facilitate population maintenancein estuarine/coastal waters.     

草原矿区土壤环境损害基线评估判定的应用研究——以内蒙古草原矿区为例

草原生态环境损害评估鉴定中,土壤基线是重要的评价标准。如何确定土壤基线,寻找适宜的基线判定方法,进而判定草原土壤的受损害程度,已成为草原生态系统损害评估鉴定、生态系统保护及修复工作中的关键。基于科学、务实、准确的原则,对内蒙古2个草原矿区土壤的总有机质（TOM）、N、P、K含量进行采样分析,采用历史数据法、参考点位法和统计点位法来判定土壤基线。结果表明,胜利矿区土壤养分均值与参考点位法基线值差异较小,采矿对胜利矿区土壤养分含量未见直接的影响。宝日希勒矿区土壤中TOM含量均值明显低于3种方法得出的基线值,N、K均值与参考点位法基线值相似,P均值位于3种基线值之间。可见采矿对宝日希勒矿区土壤中TOM的含量造成了直接影响,对N、P、K含量未见直接的影响。最适宜该地区的方法取决于样本质量和评估尺度,建议以参考点位法为主,历史数据法和统计点位法作为验证工具,3种方法共同使用。     

ALKBH5/MAP3K8 axis regulates PD-L1+ macrophage infiltration and promotes hepatocellular carcinoma progression

Hepatocellular carcinoma is one of the most common malignant tumors.M6A is a novel epigenetic modification that have been emerged as vital regulators for the progression of HCC. However, the regulatory role, clinical significance and the details of the modification, such as the impact on the local tumor environment, remain largely unclear. Our study showed that ALKBH5 was highly expressed in HCC and high ALKBH5 expression predicted a worse prognosis of HCC patients. Prediction of ALKBH5 function by tissue samples and single cell sequencing Gene Set Variation Analysis. Primary CD3 + T lymphocytes and bone marrow-derived macrophages were used to evaluate the effect of ALKBH5 on immune microenvironment. The results indicated that ALKBH5 promote HCC cell proliferation, metastasis and PD-L1+macrophage recruitment. Mechanistically the results showed that ALKBH5 regulates MAP3K8 expression in a m6A dependent manner which mediates the proliferation and metastasis of HCC cells. ALKBH5 also promotes the activation of JNK and ERK pathways through upregulating MAP3K8, thus regulating the expression of IL-8 and promoting macrophage recruitment. Taken together, these data show that ALKBH5 promotes HCC growth, metastasis and macrophage recruitment through ALKBH5/MAP3K8 axis and it may serve as a potential diagnostic marker and target for treatment of HCC patients.     

中国虫草一新记录种

【目的】对一个寄生鳞翅目幼虫的虫草标本Dxhir140901进行分类鉴定。【方法】采用形态学比较和基于ITS1-5.8S-ITS2rDNA的系统发育与进化网络分析进行鉴定。【结果】形态学观察：标本的分离菌株形态显示其为典型的被毛孢属真菌,具有两型产孢结构：A型产孢细胞柱状,(1.8?6.3) μm×1.8 μm;B型产孢细胞锥形,基部柱状,向上逐渐变细无明显颈部,基部宽3?3.8 μm,长21?63 μm,颈部宽1.8?2.0 μm,菌丝末端可直接形成产孢细胞;孢子橘瓣形或卵形,(8.1?10.8) μm×(2.7?5.4) μm,具粘液,黏液层厚1.8?2.7 μm。系统发育分析结果显示该菌株与巨针线形虫草Ophiocordyceps macroacicularis聚为一支,支持率为98%,进化网络分析也支持上述结果。【结论】通过与O. macroacicularis的形态比较和分子系统学分析结果,Dxhir140901及其分离株Gzuifr-hir140901为巨针线形虫草Ophiocordyceps macroacicularis S. Ban, T. Sakane & Nakagiri的无性阶段,该种为中国新记录种。     

基于生态系统服务的山水林田湖草生态保护修复研究——以南太行地区鹤山区为例

开展山水林田湖草生态保护修复是生态文明建设的重要内容,将生态系统服务评价成果与山水林田湖草生态保护修复工程相结合,以国家第三批山水林田湖草生态保护修复试点-河南省南太行地区鹤山区为例,探究生态系统服务评价成果在区域山水林田湖草生态保护修复中的应用。结果表明,近年来鹤山区生态系统服务价值整体呈下降趋势且生态环境破坏严重。其中,2014-2017年鹤山区各土地类型二级服务ESV减少了303.95万元。研究期内,水资源供给、气体调节功能、气候调节、净化环境、水文调节、土壤保持、生物多样性和美学景观等8项生态服务价值都出现不同程度的下降;在鹤山区生态保护修复工作中需要重点关注生态系统的水文调节、净化环境、土壤保持和生物多样性等服务功能,相应的生态保护修复工程应集中在河道生态修复与湿地保护、矿山生态环境修复与土地整治、森林及生物多样性保护修复;鹤山区山水林田湖草生态保护修复工程能够提升区域生态服务价值。其中,工程实施后的生态系统服务价值总量预计能达到36407.95万元,比2017年增加7741.96万元,增长率为27%且各项生态服务价值均有提升。研究结果对鹤山区山水林田湖草生态保护修复工程的实施有一定的指导性作用。     

长白山温带森林生态系统关键要素耦合及演替过程动态模拟

认识生态系统内和生态系统间耦合机制,揭示复合生态系统功能规律,对促进我国山水林田湖草沙项目一体化修复和保护实践具有重要的意义。针对目前修复和保护工程中出现的缺乏系统性、连续性等问题,以拥有丰富生态资源的长白山温带森林生态系统为研究区域,对其关键要素"水土气生"进行耦合建模。通过分析模型的运行机理,探究重要子模块之间的相互作用以及子模块内部生态关键要素的耦合机制,并以长白山温带落叶阔叶林的组成树种和环境因素为对象构建模型参数,通过运行林窗模型1000次,得出长白山温带森林的动态演替过程。结果表明:在森林生态系统的演替过程中,"水土气生"体现为模型中有效积温、干旱天数(低于土壤凋萎点的天数)、土壤可利用氮以及可利用光,这些关键要素之间相互影响,综合决定着每棵树木的更新、生长、死亡过程。模拟结果显示在长白山温带针阔混交林的动态演替过程中0-70 a,70-170 a,170-280 a,280-400 a四个阶段分别有不同的树种组成特征,与真实演替过程比较发现模拟林具有明显的阶段性。白桦、山杨为演替先锋树种,0-70 a期间生物量共占比为55%,70 a后生物量减少最后消失;紫椴、蒙古栎、水曲柳等为过渡树种,这些树种进一步改变了生长环境。红松在170 a前生物量占比仅为3%左右,随演替的发展生物量持续增加,170-280 a期间生物量占比15%,280 a之后红松生物量占总林分的50%。该结果模拟森林动态过程符合演替规律,充分说明多关键要素"水土气生"耦合机制的合理性,对于促进生态系统尺度上多生态要素耦合的相关研究提供了科学理论基础以及方法技术。