High-speed broadband monitoring of cell viscoelasticity in real time shows myosin-dependent oscillations

Study of the dynamic evolutions of cell viscoelasticity is important as during cell activities such as cell metastasis and invasion, the rheological behaviors of the cells also change dynamically, reflecting the biophysical and biochemical connections between the outer cortex and the intracellular structures. Although the time variations of the static modulus of cells have been investigated, few studies have been reported on the dynamic variations of the frequency-dependent viscoelasticity of cells. Measuring and monitoring such dynamic evolutions of cells at nanoscale can be challenging as the measurement needs to meet two objectives inherently contradictory to each other—the measurement must be broadband (to cover a large frequency spectrum) but also rapid (to capture the time-elapsed changes). In this study, we exploited a recently developed control-based nanomechanical protocol of atomic force microscope to monitor in real time the dynamic evolutions of the viscoelasticity of live human prostate cancer cells (PC-3 cells) and study its dependence on myosin activities. We found that the viscoelasticity of PC-3 cells, followed the power law, and oscillated at a period of about 200 s. Both the amplitude and the frequency of the oscillation strongly depended on the intracellular calcium and blebbistatin-sensitive motor proteins.     

Characterization of the interactions of a bifunctional inhibitor with alpha-thrombin by molecular modelling and peptide synthesis.

A potent thrombin inhibitor, [D-Phe45, Arg47] hirudin 45-65, that contains an active site-directed sequence D-Phe-Pro-Arg-Pro, an exosite specific fragment hirudin 55-65 (H55-65) and a linker portion hirudin 49-54, was designed based on the hirudin sequence [DiMaio et al. (1990) J. Biol. Chem., 265, 21698-21798]. A three-dimensional model of the complex between the B-chain of human thrombin and the inhibitor [D-Phe45, Arg47] hirudin 45-65 was constructed using molecular modelling starting from the X-ray C alpha coordinates of the thrombin-hirudin complex and the NMR-derived structure of the thrombin-bound hirudin 55-65. The contribution of the H49-54 fragment to the thrombin-inhibitor interaction was deduced by examining a series of analogs containing single glycine substitution and analogs with reduced number of residues within the linker. The results were consistent with the molecular modelling observations i.e. the H49-54 fragment serves the role of a spacer in the binding interaction and could be replaced by four glycine residues. The studies on the interaction of the exosite-directed portion of the inhibitor with thrombin using a series of synthetic H55-65 analogs demonstrated that residues AspH55 to ProH60 play a major role in binding to human thrombin where the side chains of PheH56, IleH59 and GluH57 showed critical contributions. Molecular modelling suggested that these side chains may contribute to inter- and intramolecular hydrophobic and electrostatic interactions, respectively.     

Circ_0002984 induces proliferation,migration and inflammation response of VSMCs induced by ox-LDL through miR‑326-3p/VAMP3 axis in atherosclerosis

Atherosclerosis can result in multiple cardiovascular diseases. Circular RNAs (CircRNAs) have been reported as significant non-coding RNAs in atherosclerosis progression. Dysfunction of vascular smooth muscle cells (VSMCs) is involved in atherosclerosis. However, up to now, the effect of circ_0002984 in atherosclerosis is still unknown. Currently, we aimed to investigate the function of circ_0002984 in VSMCs incubated by oxidized low-density lipoprotein (ox-LDL). Firstly, our findings indicated that the expression levels of circ_0002984 were significantly up-regulated in the serum of atherosclerosis patients and ox-LDL-incubated VSMCs. Loss of circ_0002984 suppressed VSMC viability, cell cycle distribution and migration capacity. Then, we carried out ELISA assay to determine TNF-α and IL-6 levels. The data implied that lack of circ_0002984 obviously repressed ox-LDL–stimulated VSMC inflammation. Meanwhile, miR-326-3p, which was predicted as a target of circ_0002984, was obviously down-regulated in VSMCs treated by ox-LDL. Additionally, after overexpression circ_0002984 in VSMCs, a decrease in miR-326-3p was observed. Subsequently, miR-326-3p was demonstrated to target vesicle-associated membrane protein 3 (VAMP3). Therefore, we hypothesized that circ_0002984 could modulate expression of VAMP3 through sponging miR-326-3p. Furthermore, we confirmed that up-regulation of miR-326-3p rescued the circ_0002984 overexpressing-mediated effects on VMSC viability, migration and inflammation. Additionally, miR-326-3p inhibitor-mediated functions on VSMCs were reversed by knockdown of VAMP3. In conclusion, circ_0002984 mediated cell proliferation, migration and inflammation through modulating miR-326-3p and VAMP3 in VSMCs, which suggested that circ_0002984 might hold great promise as a therapeutic strategy for atherosclerosis.     

Spatial analysis of hepatocellular carcinoma and socioeconomic status in China from a Population-based Cancer Registry

Background: Little is known about geographic variations in liver cancer at high incident regions. We aimed to identify spatial variation of hepatocellular carcinoma (HCC) at a high-risk area in China and determine its association with socioeconomic status (SES). Methods: Based on 2299 liver cancer cases diagnosed in Haimen from 2003 to 2006, we calculated age–sex standardized incidence ratios (SIRs) and used two spatial scan statistics to determine the geographic variations in HCC. Bayesian hierarchical model was used to explore the association between HCC incidence and SES. Results: Age and sex SIRs for HCC varied from 0.54 to 1.97 for 24 townships. The eastern region of Haimen was identified to have a significantly increased risk of HCC. Fitting of a Bayesian hierarchical model linking per-capita fiscal revenue with SIRs of HCC indicated that the area with a lower revenue had a significantly higher incidence of HCC [β_log(revenue) = ?0.179, posterior 95% Bayesian credible interval (CI) = (?0.326, ?0.04)]. Conclusions: This study demonstrated substantial geographic variation in the incidence of HCC within a high-risk region, which was associated with SES. HCC control and intervention should focus on disadvantaged areas to reduce the HCC disparities.     

Molecular endpoints of Ca2+/calmodulin- and voltage-dependent inactivation of Cav1.3 channels

Ca²⁺/calmodulin- and voltage-dependent inactivation (CDI and VDI) comprise vital prototypes of Ca²⁺ channel modulation, rich with biological consequences. Although the events initiating CDI and VDI are known, their downstream mechanisms have eluded consensus. Competing proposals include hinged-lid occlusion of channels, selectivity filter collapse, and allosteric inhibition of the activation gate. Here, novel theory predicts that perturbations of channel activation should alter inactivation in distinctive ways, depending on which hypothesis holds true. Thus, we systematically mutate the activation gate, formed by all S6 segments within Ca_V1.3. These channels feature robust baseline CDI, and the resulting mutant library exhibits significant diversity of activation, CDI, and VDI. For CDI, a clear and previously unreported pattern emerges: activation-enhancing mutations proportionately weaken inactivation. This outcome substantiates an allosteric CDI mechanism. For VDI, the data implicate a “hinged lid–shield” mechanism, similar to a hinged-lid process, with a previously unrecognized feature. Namely, we detect a “shield” in Ca_V1.3 channels that is specialized to repel lid closure. These findings reveal long-sought downstream mechanisms of inactivation and may furnish a framework for the understanding of Ca²⁺ channelopathies involving S6 mutations.     

元谋干热河谷植被的类型研究 Ⅱ.群丛以下单位

本文用法瑞植物社会学学派的方法,并经主分量分析验证,对元谋干热河谷的植被进行了重点在群丛及其以下单位的系统分类研究。结果得到9个群丛以及各群丛之下的6个亚群丛、3个群丛变型和两个群丛相的一套系统单位。     

利用Ecopath模型评价鲢鳙放养对千岛湖生态系统的影响

“保水渔业”是中国控制"水华"暴发等生态灾变的措施之一，其结果存在较大的不确定性。在浙江省的新安江水库（千岛湖），"保水渔业"的实施带来了水质改善和渔业增产的双重效果。但在生态系统自组织层面，这种人工干预手段引起的生态系统结构化效应的研究，尚未真正展开。基于2008-2010年千岛湖的生态和渔业资源调查数据，应用EwE （V6.6）构建了2010年千岛湖生态系统的Ecopath模型，并将其与2004和2016年的模型进行对照，分析了千岛湖生态系统在鲢、鳙鱼放养下的变化。千岛湖生态系统在3个年份均为4个整合营养级，营养能流分布成典型的金字塔型，且营养流总量中流向碎屑的占比很大，营养级Ⅰ、Ⅱ的能量被利用得不够充分；除鲢、鳙鱼外大部分鱼类的生物量逐渐下降，浮游植物和碎屑的生物量增多；系统的初级生产力和规模得到了一定提升，但总体的能量转换效率有所降低。在一些和系统成熟度、复原力和稳定性密切相关的参数方面，总初级生产量/总呼吸量增加，Finn氏循环指数和Finn氏平均路径长度逐渐降低，3个年份的连接指数（CI）分别为0.223、0.219、0.263，系统杂食指数（SOI）分别为0.087、0.102和0.131。研究分析表明，长期的鲢、鳙鱼放养使千岛湖的食物网结构发生了较大的变动，生态系统的营养交互关系不够复杂，成熟度和稳定性有所下降。千岛湖作为一个由水库发展而来的淡水水体，食物网关系本身就较为简单。因此，未来需要避免过多的人为干扰，并基于生态学原理更系统地进行修复工作。