鲟鱼发酵过程中微生物的演替变化分析

为了解鲟鱼发酵中细菌群落多样性及动态演替规律,为发酵鲟鱼的品质提供理论数据,利用Illumina HiSeq测序平台,采集鲟鱼发酵过程中的7个阶段的样品,对样品中的细菌16S rDNA V4~V5区进行测序和系统发育分析。结果表明,样品获得序列数平均为46 463条,各个发酵阶段的操作分类单元数目分别为新鲜鱼样1 064,腌制鱼样952;发酵5 d 454,发酵10 d 442,发酵20 d 327,发酵25 d 372和发酵35 d 356。7组样品中总体细菌组成较为复杂,含有38个门、196个科,在新鲜鱼样中,厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)占主要优势;在腌制阶段,蓝藻门(Cyanobacteria)占主要优势;从发酵开始至结束,厚壁菌门(Firmicutes)占绝对优势。在科水平上,新鲜鱼样中的主要优势菌科为肠杆菌科(Enterobacteriaceae),腌制样品中的主要优势菌科为肠球菌科(Enterococcaceae);从发酵开始至结束,主要的优势菌科为明串珠菌科(Leuconostocaceae),其次是乳杆菌科(Lactobacillaceae)和链球菌科(Streptococceae)。通过HiSeq测序能够更全面解析鲟鱼发酵过程中的细菌多样性;从测序平台提供的微生物多样性信息中发现,各样品间的菌属丰度存在一定的差异性,说明菌群组成与发酵环境密切相关,这为传统腌鱼发酵提供了理论依据和数据支撑。     

Phragmites stimulation and natural attenuation of indigenous microbial community accelerated bioremediation of oil contaminated soil

Illumina high-throughput sequencing method was used to reflect fluctuation of indigenous microbial community during phragmites stimulation process in this study. Results showed Proteobacteria, Fimicutes, and Actinobacteria were most influential in process, Firmicutes and Deinococcus-Thermus were positively affected by nutrient amendment. Firmicutes can be seen as indicator of degradation rate. Dietzia, Salinimicrobium, and Halomonas were the most dominant genus in both Phragmites and Blank setups. These results indicated phragmites addition improved the efficiency of biodegradation and significantly regulated the microbial community structure and functions during remediation process.     

Identification of Potent Caspase-8 Inhibitors from a Library of Fluorescent Natural Products Screened by an AIEgen-Based Light-Up Probe

Fluorescent natural products are a rich source of drugs and chemical probes, but their innate fluorescence can interfere with fluorescence-based screening assays. Caspase-8 is a key player in apoptosis, its inhibition having been found to be beneficial for treatment of inflammatory and neurodegenerative diseases. Small-molecular inhibitors of caspase-8 remain sparsely reported, however. In this study, we firstly developed a light-up probe based on an AIEgen and capable of targeting caspase-8. This fluorescent dye has a Stokes shift of 200 nm, which could allow the innate fluorescence signals of natural products to be avoided. On screening a library of 86 fluorescent natural products, we found for the first time that gossypol showed potent inhibition of caspase-8 in vitro and in situ. This unique light-up probe, coupled with colored natural products, could represent an efficient approach to hit discovery for druggable targets.     

常温常压等离子体诱变选育高产L-异亮氨酸谷氨酸棒杆菌

以谷氨酸棒杆菌（Corynebacterium glutamicum）23798为原始菌株,对其进行常温常压等离子体（ARTP）诱变,以磺胺胍抗性和氨基酸与茚三酮特异显色为筛选标记,以期得到高产L-异亮氨酸的诱变谷氨酸棒杆菌,并对其遗传稳定性进行研究。结果表明,原始菌株23798经过ARTP诱变处理180 s后,经0.4 mg/mL磺胺胍抗性筛选、多孔板高通量筛选、发酵培养复筛,选育出一株高产L-异亮氨酸诱变谷氨酸棒杆菌（Corynebacterium glutamicum）B1。该菌株在摇瓶中发酵培养48 h,L-异亮氨酸产量达18.5 g/L,比原始菌株提高62.03%,且遗传性状稳定。     

酱香型白酒第四轮次酒酿造过程中真菌多样性分析

该研究应用Illumina MiSeq高通量测序技术解析酱香型白酒第四轮次酒酿造过程中真菌多样性,并阐明其优势真菌群落结构及其随酿造工艺动态变化。结果表明,在酒曲中主要优势菌有曲霉属（Aspergillus）、嗜热子囊菌属（Thermoascus）;堆积发酵酒醅中主要优势菌有拟青霉属（Paecilomyces）、念珠菌属（Candida）;窖内发酵酒醅中主要优势菌有嗜热子囊菌属（Thermoascus）、嗜热真菌属（Thermomyces）;窖泥中主要优势菌有假霉样真菌属（Pseudallescheria）、镰刀菌属（Fusarium）。同一酒厂和不同酒厂新老车间酒曲、堆积发酵、窖内发酵、窖泥之间真菌组成相似度较高,但其优势菌群丰度差异显著。发酵车间使用年限及窖龄影响着酿酒微生物多样性;使用时间长的车间和窖池其环境微生物种群结构更稳定,优势菌群更突出。     

镇巴腊肉生产过程中微生物组成及多样性变化

基于高通量测序技术,对镇巴腊肉加工过程不同阶段的微生物种群组成进行鉴定和分析,同时对细菌表型及相关功能进行预测。结果表明：镇巴腊肉生产加工过程中,真菌丰度大于细菌丰度,并随着发酵时间及加工工艺变化,微生物群落存在明显的演替;镇巴腊肉中丰度最高的细菌为厚壁菌门（Firmcutes）和变形菌门（Proteobacteria）,优势细菌属随着加工步骤推进由原料肉样品中的大肠-志贺氏菌属演替为腌制时期和熏制时期的葡萄球菌属（Staphylococcus）、嗜冷杆菌属（Psychrobacter）和乳杆菌属（Latilactobacillus）;丰度最高的真菌门为子囊菌门（Ascomycota）和担子菌门（Basidiomycota）,优势真菌属由镰刀菌属（Fusarium）和被孢霉属演替为镰刀菌属（Mortierella）和德巴利氏酵母属（Debaryomyces）;基于菌落演替变化及细菌表型和功能预测,发现镇巴腊肉在加工过程中有害微生物不断减少,利于发酵的微生物逐渐增加,促进镇巴腊肉风味物质形成。     

基于色谱-质谱联用技术的食品中农药残留高通量非靶向检测技术研究进展

农药残留导致的食品安全事件威胁人们的健康和生命。现阶段食品中的农药残留具有品类多、残留量低且施用未知等特点,检测难度较大。因此,开发食品中农药残留高通量非靶向检测技术具有重要意义。色谱-质谱联用技术因具有高分离、高鉴别、高通量、非靶向等优点,在农药残留检测领域得到了广泛应用。本文对近10年国内外基于色谱-质谱联用技术的食品中农药残留高通量非靶向检测相关报道进行归纳和总结,简要介绍食品中多农药残留检测的现状,重点从多维色谱及高分辨质谱技术的开发、吸附材料及样品前处理技术的改进、化学计量学方法的优化等方面阐述色谱-质谱联用技术在食品中农药残留检测中的应用进展,并对其存在的问题提出相关建议,以期为色谱-质谱联用技术在食品中多农药残留检测领域的相关研究应用提供新的参考依据。     

浙江地区中温大曲贮存过程细菌群落多样性及功能预测分析

利用16S rRNA高通量测序和理化分析技术分别解析浙江地区中温大曲贮存过程中细菌群落多样性和理化特征演替,并结合生物信息学方法预测菌群功能。结果表明,大曲贮存90 d后,pH和含水量显著降低（P<0.05）,酸度显著升高（P<0.05）,且贮存前后细菌群落多样性和结构组成差异显著（P<0.05）。乳酸杆菌属（Lactobacillus）和魏斯式菌属（Weissella）是大曲贮存期间的绝对优势属,其相对丰度范围分别为47.56%～63.44%和25.16%～42.77%。魏斯式菌属相对丰度与含水量和pH呈正相关,而酸度与乳酸杆菌属、醋酸杆菌属（Acetobacter）和克罗彭斯特菌属（Kroppenstedtia）相对丰度呈正相关。此外,功能预测结果表明中温大曲贮存期间的基因功能主要与代谢、遗传信息处理和环境信息处理等功能相关。     

高通量测序分析新疆不同产区葡萄酒对大鼠肠道菌群多样性的影响

将雄性SD大鼠随机分为5组（A1～A5）,分别灌胃新疆焉耆盆地、伊犁河谷、天山北麓、吐哈盆地葡萄酒、蒸馏水8周后,收集粪便,采用高通量测序技术分析大鼠肠道菌群变化,探讨不同产区葡萄酒对大鼠肠道菌群的影响。结果表明,各组大鼠肠道菌群具有一定的差异,主要共有优势菌门为厚壁菌门（Firmicutes）和拟杆菌门（Bacteroidetes）,优势菌属为乳杆菌属（Lactobacillus）,A1～A4组中主要优势菌门和菌属的相对丰度之和均比A5组显著升高（P<0.05）,其中A2组升高幅度最大,其有害菌门变形菌门（Proteobacteria）、脱硫菌门（Desulfobacterota）相对丰度显著降低（P<0.05）;有益菌属罗姆布茨菌属（Romboutsia）、艾克蔓菌属（Akkermansia）及乳杆菌属（Lactobacillus）相对丰度显著增加（P<0.05）。结合多级物种差异判别分析,A2组肠乳杆菌（Lactobacillus intestinalis）的富集最具差异性（P<0.05）。说明适量摄入伊犁河谷干红葡萄酒一定程度上能改善肠道菌群。     

A Microfluidic 3D Endothelium-on-a-Chip Model to Study Transendothelial Migration of T Cells in Health and Disease

The recruitment of T cells is a crucial component in the inflammatory cascade of the body. The process involves the transport of T cells through the vascular system and their stable arrest to vessel walls at the site of inflammation, followed by extravasation and subsequent infiltration into tissue. Here, we describe an assay to study 3D T cell dynamics under flow in real time using a high-throughput, artificial membrane-free microfluidic platform that allows unimpeded extravasation of T cells. We show that primary human T cells adhere to endothelial vessel walls upon perfusion of microvessels and can be stimulated to undergo transendothelial migration (TEM) by TNFα-mediated vascular inflammation and the presence of CXCL12 gradients or ECM-embedded melanoma cells. Notably, migratory behavior was found to differ depending on T cell activation states. The assay is unique in its comprehensiveness for modelling T cell trafficking, arrest, extravasation and migration, all in one system, combined with its throughput, quality of imaging and ease of use. We envision routine use of this assay to study immunological processes and expect it to spur research in the fields of immunological disorders, immuno-oncology and the development of novel immunotherapeutics.