基于深度学习的秀丽隐杆线虫显微图像分割方法

利用计算机实现自动、准确的秀丽隐杆线虫(C.elegans)的各项形态学参数分析,至关重要的是从显微图像上分割出线虫体态,但由于显微镜下的图像噪声较多,线虫边缘像素与周围环境相似,而且线虫的体态具有鞭毛和其他附着物需要分离,多方面因素导致设计一个鲁棒性的C.elegans分割算法仍然面临着挑战。针对这些问题,提出了一种基于深度学习的线虫分割方法,通过训练掩模区域卷积神经网络(Mask R-CNN)学习线虫形态特征实现自动分割。首先,通过改进多级特征池化将高级语义特征与低级边缘特征融合,结合大幅度软最大损失(LMSL)损失算法改进损失计算;然后,改进非极大值抑制;最后,引入全连接融合分支等方法对分割结果进行进一步优化。实验结果表明,相比原始的Mask R-CNN,该方法平均精确率(AP)提升了4.3个百分点,平均交并比(mIOU)提升了4个百分点。表明所提出的深度学习分割方法能够有效提高分割准确率,在显微图像中更加精确地分割出线虫体。     

A Neural Network Model of Caenorhabditis Elegans: The Circuit of Touch Sensitivity

The paper presents a neural network model of the touch sensitivity circuit of the nematode Caenorhabditis elegans. We describe a serie of simulations in which neural networks are trained, using a genetic algorithm, to reproduce the habituation of the nematode's touch sensitive behavior. A lesion study of the network allows to make a direct comparison between the fine functioning of the model and the data collected in real organisms. The model accords well with the known neurobiological data and it suggests some hypotheses about the functioning of the neural circuit and of single neurons.     

Short communication: Development of a direct in vivo screening model to identify potential probiotic bacteria using Caenorhabditis elegans

Caenorhabditis elegans is an accepted model host to study host-bacteria interactions in the gut, in addition to being a simple model with which to study conserved aspects of biological signaling pathways in intestinal environments, because these nematode worms have similar intestinal cells to those of humans. Here, we used C. elegans to develop a new in vivo screening system for potential probiotic lactic acid bacteria (LAB). Initially, critical colonization ability of LAB strains isolated from Korean infant feces was screened in the worm intestinal tract over a period of 5 d. Furthermore, we investigated host health-promoting activities, including longevity-extending effects and immune-enhancing activities against foodborne pathogen infection. We identified 4 LAB strains that were highly persistent in the nematode gut and that significantly prolonged the longevity of C. elegans and improved the survival of C. elegans in response to infection by Staphylococcus aureus. The 4 LAB strains we identified showed resistance to acid and bile conditions, assimilated cholesterol, and were able to attach to a mucus layer. The 4 LAB isolates were identified as Lactobacillus plantarum using 16S rRNA sequencing analysis. Taken together, we developed a direct in vivo screening system using C. elegans to study host health-promoting LAB. Our system is simple, rapid, cost-effective, and reliable, and we anticipate that this system will result in the discovery of many more potential probiotic bacteria for dairy foods.     

绿春地区断肠草生物总碱提取工艺优化

优选微波法提取断肠草生物总碱的工艺,以断肠草生物总碱的提取率为指标,考察不同提取条件下的提取率。先通过单因素实验对断肠草生物总碱的提取工艺中溶剂种类、提取系统中固液比、浸提时间、浸提温度4个因素进行优选,进而考察多因素协同作用对生物总碱提取率的影响设计了L9（33）的正交实验。结果表明,采用氯仿作为提取溶剂、微波功率500W、固液比1∶8（g/L）、浸提时间240 s（80 s×3）、浸提温度55℃时生物总碱得率最高为1.63%,因此,以氯仿为提取溶剂,经微波辅助技术处理后生物总碱提取量显著得到了提高。     

Glucosamine Extends the Lifespan of Caenorhabditis elegans via Autophagy Induction

Glucosamine (GlcN) is commonly used as a dietary supplement to promote cartilage health in humans. We previously reported that GlcN could induce autophagy in cultured mammalian cells. Autophagy is known to be involved in the prevention of various diseases and aging. Here, we showed that GlcN extended the lifespan of the nematode Caenorhabditis elegans by inducing autophagy. Autophagy induction by GlcN was demonstrated by western blotting for LGG-1 (an ortholog of mammalian LC3) and by detecting autophagosomal dots in seam cells by fluorescence microscopy. Lifespan assays revealed that GlcN-induced lifespan extension was achieved with at least 5 mM GlcN. A maximum lifespan extension of approximately 30 % was achieved with 20 mM GlcN (p<0.0001). GlcN-induced lifespan extension was not dependent on the longevity genes daf-16 and sir-2.1 but dependent on the autophagy-essential gene atg-18. Therefore, we suggest that oral administration of GlcN could help delay the aging process via autophagy induction.     

A Preliminary Study of the Application of a Model Animal-Caenorhabidity elegans＇ Exposure to a Low-Energy Ion Irradiation System

Because of the lack of suitable animal models adapted to high vacuum stress in the low-energy ion implantation system, the bio-effects ion irradiation with an energy less than 50 keV on multi-cellular animal individuals have never been investigated so far. The nematode Caenorhabditis elegans has proved to be an excellent animal model used for the study of a broad spectrum of biological issues. The purpose of this work was to investigate the viability of this animal under ion irradiation. We studied the protection effects of glycerol and trehalose on the enhancement of nematodes＇ ability to bear the vacuum stress. The results showed that the survival of the nematodes was enhanced remarkably under long and slow desiccation, even without glycerol and trehalose. 159 glycerol showed a better anti-vacuum stress effect on the nematodes than trehalose did under short-time desiccation. Low-temperature pre-treatment or post-treatment of the samples had no obvious effect on the survival scored after argon ion irradiation. Moreover, little effect was induced by 15% glycerol- and vacuum-exposure on germ cell apoptosis, compared to the untreated control sample. It issuggested that such treatment would provide relatively low background for genotoxic evaluations with ion irradiation.     

铁皮石斛多糖调控线虫寿命及抗氧化作用的研究

摘 要: 目的 基于秀丽隐杆线虫自然衰老模型, 探讨铁皮石斛多糖(Dendrobium candidum polysaccharides, DOP)抗衰老作用及其机制。方法 将秀丽隐杆线虫野生型N2随机分为对照组和铁皮石斛多糖组(低、中、高), 观察铁皮石斛多糖对线虫N2寿命、运动能力和咽泵能力的影响; 通过检测氧化应激、抗氧化酶试剂盒以及活性氧评估铁皮石斛多糖对N2线虫抗氧化能力的影响。基于skn-1基因缺陷型突变株(EU1)线虫及带有荧光标记的转基因线虫CL2166分析铁皮石斛多糖延长健康寿限的可能作用途径; 采用实时荧光定量聚合酶链反应技术测定铁皮石斛多糖作用下线虫抗氧化、胰岛素信号途径相关基因转录水平。结果 铁皮石斛多糖中剂量组(250 μg/mL)可显著延长线虫N2的寿命, 平均延寿率达12.73%, 可显著提高运动、咽泵、抗氧化应激能力; 显著提高过氧化氢酶(catalase, CAT)、超氧化物歧化酶(superoxide disumutase, SOD)活性和谷胱甘肽(glutathione, GSH)含量, 降低活性氧(reactive oxygen species, ROS)水平和丙二醛(malondialdehyde, MDA)含量; 铁皮石斛多糖干预处理对skn-1基因缺陷线虫的延寿效果缺失; 但能够增强CL2166线虫的荧光强度; 同时上调sod-5、sod-3、ctl-1、ctl-2、daf-16和skn-1基因的表达水平, 下调daf-2基因的表达水平。结论 铁皮石斛多糖可能通过激活Nrf2/SKN-1和DAF-16途径拮抗氧化应激从而延缓线虫衰老。     

铁皮石斛叶多糖对秀丽隐杆线虫体内抗衰老作用

目的：基于模式生物秀丽隐杆线虫（以下简称线虫）探究铁皮石斛叶多糖的抗衰老能力。方法：采用含不同质量浓度（0.5、1.0、2.0 mg/mL）铁皮石斛叶多糖的大肠埃希氏菌OP50菌液每天饲喂线虫,以饲喂大肠埃希氏菌OP50菌液组作为空白对照,观察铁皮石斛叶多糖对线虫寿命、生殖能力、运动能力和应激能力的影响,同时测定线虫体内活性氧水平及抗氧化酶的酶活力,分析铁皮石斛叶多糖对线虫体内抗氧化能力的影响。结果：与空白对照组线虫相比,在本实验考察的质量浓度范围内,铁皮石斛叶多糖均可延长线虫的寿命,提高其运动、应激能力,同时不会损害线虫的生殖能力,并且能有效提高过氧化氢酶和超氧化物歧化酶的活力,显著降低线虫体内丙二醛含量（P＜0.05）。结论：铁皮石斛叶多糖可能通过提高线虫抗氧化酶活力及提高线虫抗应激能力,降低线虫体内活性氧水平并减缓细胞氧化损伤,进而发挥延长线虫寿命的功效。