Three‐dimensional ring‐oven washing technique for a paper‐based immunodevice

Washing is a standard step for enzyme‐linked immunosorbent assays (ELISA) performed on a paper‐based chip, in which nonspecific‐binding antibodies and antigens should be removed completely from the paper surface. In this study, a novel three‐dimensional (3D) washing strategy using a heating ring‐oven was carried out on a paper‐based chip. Compared with a plane washing mode by a ring‐oven, this 3D washing strategy obtained a lower background, as gravity played an important role in the washing step. The paper‐based chip was placed on a 3D plastic holder and the waste area was connected to a heating ring. Use of a heating waste area meant that the nonspecific‐binding protein was continuously carried to the waste area through gravity and capillary action. The angle between the plastic holder and the ring plane was carefully selected. The effect of washing on different parts of the detection area was investigated by upconversion fluorescence and chemiluminescence (CL). This novel 3D washing strategy was performed for carcinoembryonic antigen detection through CL and a lower detection limit of 2 pg ml^?1 was obtained. This approach provides an effective washing strategy to remove nonspecific‐binding antibody from a paper‐based immunodevice.     

The enantiomer pair of 24S- and 24R-hydroxycholesterol differentially alter activity of large-conductance Ca2+-dependent K+ (slo1 BK) channel

24S-hydroxycholesterol (HC) is most abundant oxysterols in the brain, passes through blood brain barrier, and is therefore regarded as an intermediary for brain cholesterol elimination. We reported that large-conductance Ca²⁺- and voltage-activated K⁺ (slo1 BK) channels are suppressed by this oxysterol, which is presumably intercalated into cell membrane to access the outer surface of the channel. Such an outer approach would make it difficult to interact with the inner, ion-conducting part of the channel. The present findings showed that 24R-HC, the racemic counterpart of 24S-HC, also suppressed slo1 BK channel but in a different voltage-dependent manner. There was a difference between the effects of the two enantiomers on activation kinetics but not on deactivation kinetics. It is suggested that the chirality contributes to the efficacy of channel blockers that act from outer lipophilic parts of channels, as with those which act on the inner, ion-permeable surface.     

土壤水分对黄土区苹果园土壤-植物-大气连续体(SPAC)中水势梯度的影响

开展土壤-植物-大气连续体(SPAC)中水势变化的联动分析对于揭示植物水分状况对环境变化的响应机制有重要意义。在黄土高原苹果园对果树生长季大气水势(Ψ_air)、木质部水势(Ψ_stem)、土壤水势(Ψ_soil)进行连续监测与分析。结果表明: 苹果树在生长季的日平均Ψ_stem为-0.57 MPa,在-0.24～-2.0 MPa间变化。苹果树SPAC连续体中水势梯度(Ψ_soilΨ_stemΨ_air)平均为1∶9.8∶1155,其中植-土界面的水势梯度(Ψ_stemΨ_soil)与土壤体积含水率(VWC)间呈极显著的线性正相关关系,Ψ_stem与Ψ_soil间呈良好的线性关系,且相关性强于Ψ_stem与Ψ_air间的相关性。当Ψ_soil<-0.08 MPa(VWC=17%,约为田间持水量的0.56倍)时,Ψ_stem对Ψ_soil变化响应的敏感性明显降低,气-植界面的水势梯度(Ψ_airΨ_stem)与Ψ_soil间由无明显关系转变为紧密的线性相关关系(R²=0.93)。Ψ_air对Ψ_stem的驱动存在阈值效应,在Ψ_air降到-69 MPa前,Ψ_stem日变化幅度随Ψ_air增加而增加,之后呈下降趋势。土壤含水率降低引起苹果树水势、SPAC各界面上的水势梯度明显下降,且在土壤含水率降至约17%时存在阈值效应。研究结果为理解树木水分状况对土壤、大气干旱的响应机制提供了重要依据。     

亚热带-温带气候过渡区落叶阔叶林物种-生境关联分析

物种-生境关联分析有利于更好地理解物种共存理论和群落构建机制。根据秦岭落叶阔叶林25 hm²固定监测样地的调查数据,将树种分为幼苗、幼树和成树3个生活阶段,利用Torus-translation检验方法分析物种与不同生境类型之间的关联性。结果表明: 生境对各物种的影响不同。与高坡显著关联的物种数最多,其中95.7%为负关联;与低坡呈负相关的物种占89.5%;与山脊呈显著负关联的物种占90.9%;物种与高谷生境多存在显著正关联,呈负相关的只有1种,占0.03%。物种在幼苗、幼树和成树阶段与生境分别存在80、44和23个关联,表明幼苗阶段对生境的依赖程度更大。幼苗阶段的物种中有38个(占总物种数的90.5%)至少与一类生境存在显著的关联性;幼树阶段有25个(占总物种数的58.1%)至少与一类生境存在显著关联;成树阶段只有17个(占总物种数的39.5%)至少与一类生境存在显著关联。同一生境对不同生活史阶段物种的影响存在差异,到生活史阶段后期,生境的影响逐渐减弱。由于特定的环境需求,多数物种在不同生活史阶段表现出不同的生境偏好。     

鄱阳湖流域极端降水时空分布和非平稳性特征

全球变暖背景下的极端天气气候事件显著增加。本研究基于PreWhitening Mann-Kendall(PWMK)、极点对称模态分解法和广义可加模型,利用鄱阳湖流域1959—2019年16个国家级气象站点的逐日降水数据,从极端降水的强度、频率和持续性3个维度系统检测和分析流域极端降水的时空分布和非平稳性特征。结果表明: 研究期间,鄱阳湖流域极端降水强度和频率呈显著增加趋势,持续性呈下降趋势,极端降水整体表现出强度大、频率高、持续时间短的特点;极端降水存在明显的汛期和非汛期时间分异规律,汛期极端降水集中在流域北部和中部,而非汛期多集中于中部,子流域中信江流域降水量增加趋势最显著,达到2.10 mm·a^-1;汛期极端降水的持续时间越长,强度和范围越小,非汛期极端降水则相反;鄱阳湖流域的极端降水强度和频率以平稳性特征为主,持续性表现出非平稳性特征。随着鄱阳湖流域极端降水量的不断增加,其可能引发的灾害风险将进一步增大。     

Leptin gene-targeted editing in ob/ob mouse adipose tissue based on the CRISPR/Cas9 system

Gene therapy has become the most effective treatment for monogenic diseases. Congenital LEPTIN deficiency is a rare autosomal recessive monogenic obesity syndrome caused by mutations in the Leptin gene. Ob/ob mouse is a monogenic obesity model, which carries a homozygous point mutation of C to T in Exon 2 of the Leptin gene. Here, we attempted to edit the mutated Leptin gene in ob/ob mice preadipocytes and inguinal adipose tissues using CRISPR/Cas9 to correct the C to T mutation and restore the production of LEPTIN protein by adipocytes. The edited preadipocytes exhibit a correction of 5.5% of Leptin alleles and produce normal LEPTIN protein when differentiated into mature adipocytes. The ob/ob mice display correction of 1.67% of Leptin alleles, which is sufficient to restore the production and physiological functions of LEPTIN protein, such as suppressing appetite and alleviating insulin resistance. Our study suggests CRISPR/Cas9-mediated in situ genome editing as a feasible therapeutic strategy for human monogenic diseases, and paves the way for further research on efficient delivery system in potential future clinical application.