含半胱氨酸的天冬氨酸蛋白水解酶(caspase-1)对猪伪狂犬病毒复制的影响

猪伪狂犬病是伪狂犬病毒(Pseudorabies virus,PRV)感染引起的一种烈性接触性传染病,其感染宿主会触发机体先天免疫应答,引起I型干扰素(Type I interferon,IFN-1)和炎性细胞因子等细胞因子的产生,为研究可诱导产生炎性细胞因子的含半胱氨酸的天冬氨酸蛋白水解酶(Cysteinyl aspartate specific proteinase 1,caspase-1)的基因敲除对PRV复制的影响,本试验利用近年来发展迅速的一项规律性短重复回文序列簇/Cas9核酸酶(Clustered regulatory interspaced short palindromic repeat/CRISPR associated system 9,CRISPR/Cas9)基因定点修饰技术构建猪肾上皮细胞(Porcine kidney epithelial cells,PK15)caspase-1基因稳定敲除细胞系,并通过T7核酸酶检测敲除效率;细胞毒性(Cell counting kit-8,CCK-8)试剂盒检测PK15敲除caspase-1增殖影响;采用流式细胞术检测PRV-GFP感染PK15以及PK15-caspase-1^-/-的增殖差异;实时荧光定量PCR(Real-time quantitative PCR,RT-PCR)检测PRV-gB、TK及白细胞介素1β(Interleukin-1β,IL-1β)、IFN-β、干扰素刺激基因(Interferon-stimulated genes 20,ISG20)mRNA的表达;Western Blot检测PRV-gB蛋白表达;滴度测定检测子代病毒滴度。结果表明,2对特异性单链引导RNA(Single guide RNA,sgRNA)均能对caspase-1进行基因编辑,但经T7核酸酶酶切进行基因编辑效率分析结果表明sgRNA2的基因编辑效率较高;CCK-8试剂盒检测细胞活力结果表明caspase-1基因敲除对PK15以及PK15-caspase-1^-/-细胞活力无影响(P>0.05);流式细胞仪检测结果表明PRV-GFP在PK15-caspase-1^-/-中的增殖显著低于PK15细胞(P<0.05);定量RT-PCR结果表明PRV-gB、TK基因在PK15-caspase-1^-/-的mRNA表达显著低于PK15细胞(gB:P<0.05,TK:P<0.05),而IFN-β、ISG20基因在PK15-caspase-1^-/-的mRNA表达显著高于PK15细胞(gB:P<0.05,TK:P<0.05);Western Blot结果表明,PRV的gB蛋白在PK15-caspase-1^-/-的表达显著低于PK15细胞(P<0.05);滴度测定结果表明,敲除caspase-1能够抑制PRV子代病毒的增殖。以上结果均表明caspase-1基因敲除可抑制PRV在PK15细胞中复制。     

MiR319-targeted OsTCP21 and OsGAmyb regulate tillering and grain yield in rice

Multiple genes and microRNAs (miRNAs) improve grain yield by promoting tillering. MiR319s are known to regulate several aspects of plant development; however, whether miR319s are essential for tillering regulation remains unclear. Here, we report that miR319 is highly expressed in the basal part of rice plant at different development stages. The miR319 knockdown line Short Tandem Target Mimic 319 (STTM319) showed higher tiller bud length in seedlings under low nitrogen (N) condition and higher tiller bud number under high N condition compared with the miR319a-overexpression line. Through targets prediction, we identified OsTCP21 and OsGAmyb as downstream targets of miR319. Moreover, OsTCP21 and OsGAmyb overexpression lines and STTM319 had increased tiller bud length and biomass, whereas both were decreased in OsTCP21 and OsGAmyb knockout lines and OE319a. These data suggest that miR319 regulates rice tiller bud development and tillering through targeting OsTCP21 and OsGAmyb. Notably, the tiller number and grain yield increased in STTM319 and overexpression lines of OsTCP21 and OsGAmyb but decreased in OE319a and knockout lines of OsTCP21 and OsGAmyb. Taken together, our findings indicate that miR319s negatively affect tiller number and grain yield by targeting OsTCP21 and OsGAmyb, revealing a novel function for miR319 in rice.     

Advancement and properties of circular RNAs in prostate cancer: An emerging and compelling frontier for discovering

Prostate cancer (PC) is the most common carcinoma among men worldwide which results in 26% of leading causes of cancer-related death. However, the ideal and effective molecular marker remains elusive. CircRNA, initially observed in plant-infected viruses and Sendai virus in 1979, is generated from pre-mRNA back-splicing and comes in to play by adequate expression. The differential expression in prostate tissues compared with the control reveals the promising capacity in modulating processes including carcinogenesis and metastasis. However, the biological mechanisms of regulatory network in PC needs to systemically concluded. In this review, we enlightened the comprehensive studies on the definite mechanisms of circRNAs affecting tumor progression and metastasis. What''s more, we validated the potential clinical application of circRNAs serving as diagnostic and prognostic biomarker. The discussion and analysis in circRNAs will broaden our knowledge of the pathogenesis of PC and further optimize the current therapies against different condition.     

贵州花溪大学城破碎化林地鸟类多样性与嵌套分布格局

为揭示城镇化进程中生境破碎化对鸟类多样性及分布格局的影响, 本研究于2017-2019年每年的4-8月使用样线法对贵州花溪大学城26块破碎化林地(面积介于0.3-290.4 ha)中的鸟类群落进行了10次调查。共记录到鸟类78种, 隶属于11目37科。其中, 东洋界物种数占56.4%, 古北界物种数占32.1%, 广布种占11.5%; 有中国特有种1种。剔除高空飞行、非森林鸟类及偶然出现物种后, 不同斑块中的鸟类物种数介于12-55之间, 平均每个斑块有23.2 ± 10.5种。线性回归分析显示, 鸟类物种丰富度与林地斑块的面积有显著相关性, 斑块面积越小, 鸟类物种丰富度越低; 斑块隔离度对物种丰富度没有显著影响。基于物种多度分布矩阵的WNODF (weighted nestedness metric based on overlap and decreasing fill)嵌套分析显示, 不同斑块中鸟类群落呈现出反嵌套结构。小斑块中鸟类物种丰富度较低可能与植物丰富度较低、食物资源稀缺和繁育条件不足有关, 但短距离的隔离对鸟类迁入或扩散影响有限。环境过滤效应、种间竞争或优先效应可能导致不同斑块间存在较大的物种组成差异, 从而导致反嵌套格局。因此, 本研究建议在城市规划建设中应注重维持栖息地的完整性, 对不同面积大小的破碎化斑块都应加以保护。     

隐匿性胫骨平台骨折MRI、CT检查的影像学表现及其诊断价值对比的回顾性研究

目的:研究对比隐匿性胫骨平台骨折(TPOF)磁共振成像(MRI)、电子计算机断层扫描(CT)检查的影像学表现及其诊断价值。方法:回顾性分析我院自2016年1月至2019年12月拟诊断为TPOF且X线检查表现为阴性的89例患者的临床资料,分别对所有受试者进行MRI、CT检查,且以手术检查为金标准,比较上述两种影像学检查手段诊断TPOF的效能。此外,比较MRI、CT检查诊断TPOF的表观扩散系数以及节段各向异性值以及对TPOF类型的检出率。结果:MRI检查诊断TPOF的灵敏度、特异度及准确度分别为98.61%、94.12%、97.75%,均高于CT检查的79.17%、64.71%、76.40%(均P<0.05)。MRI检查诊断TPOF的表观扩散系数高于CT检查,而节段各向异性值低于CT检查(均P<0.05)。MRI检查对骨皮质骨折的检出率低于CT检查,而对骨小梁骨折的检出率高于CT检查(均P<0.05)。结论:MRI检查诊断TPOF的价值高于CT检查,且在骨小梁骨折的检出率方面优于CT检查,但CT检查应用于骨皮质骨折的诊断价值更高。临床工作中可能通过联合MRI以及CT检查,继而达到提高TPOF检出率的目的。     

盐土生古菌属（Haloterrigena）分类研究进展

盐土生古菌属(Haloterrigena)是一类生长在含有1.7～5.5 mol/L NaCl、17～60℃温度范围内的极端嗜盐古菌。自该属1999年被建立以来,目前已有10个种被有效发表。但是在基于16S rRNA或rpoB'基因的系统进化树上,盐土生古菌属物种常与碱线菌属(Natrinema)和嗜盐碱红菌属(Natronorubrum)的成员交叉聚类在一起;而且盐土生古菌属内各成员间的特征也不尽相同,如多数盐土生古菌属菌株生长pH范围6.0～9.0,最适pH 7.0～7.5,然而大庆盐土生古菌(Haloterrigena daqingensis)为该属的唯一嗜碱成员,生长pH在8.0～10.5,最适为pH 10.0;在化学分类特征方面,该属的多数菌株含有糖脂类型为双硫化二糖基二醚(disulfated diglycosyl diether,S2-DGD-1),然而伊斯帕尼亚盐土生古菌(Haloterrigena hispanica)菌株FP1含有S-DGD硫酸化二糖基二醚(sulfated diglycosyl diether)。鉴于该属存在的这些特殊特征,以及与碱线菌属和嗜盐碱红菌属在系统进化上的特殊关系,结合前期发表的该属的新种和相关文献资料,系统综述了盐土生古菌属的建立、分类学特征、生态分布、基因组信息等方面的研究进展,以期为盐土生古菌属已知种分类地位的确定、该属其他新分离菌株的分类鉴定、功能基因资源的挖掘与开发提供借鉴。     

Biosynthesis,characterization and antimicrobial activity of silver nanoparticles by a halotolerant Bacillus endophyticus SCU-L

Abstract

We have conducted a thorough study on extracellular biosynthesis of silver nanoparticles (AgNPs) by a halotolerant bacterium Bacillus endophyticus SCU-L, which was identified by 16S rRNA gene sequencing analysis. This strain was selected during an ongoing research programme aimed at finding a novel biological method for green nanosynthetic routes using the extremophiles in unexplored hypersaline habitats. The biosynthesized AgNPs were characterized and analyzed with UV–vis spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, atomic force microscopy and X-ray diffraction. Further, the AgNPs were found to be spherical in shape with an average particle size of about 5.1?nm, and it was stable in aqueous solution for three months period of storage at room temperature under dark condition. Also, the synthesized AgNPs significantly presented antimicrobial activity against Candida albicans, Escherichia coli, Salmonella typhi and Staphylococcus aureus. The above results suggested that the present work may provide a valuable reference and theoretical basis for further exploration on microbial biosynthesis of AgNPs by halotolerant bacteria.     

piRNA-Triggered MIWI Ubiquitination and Removal by APC/C in Late Spermatogenesis

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Highlights? MIWI is a substrate of APC/C, and piRNA loading is essential for MIWI ubiquitination ? piRNA loading promotes MIWI binding to the APC/C substrate-binding subunit ? MIWI and piRNAs are coordinately eliminated in late spermatids ? Inhibition of MIWI destruction in late spermatids prevents sperm maturation     

Phylogenetic position of Jaoa,a green algal genus endemic to China

Jaoa prasina, a freshwater green alga endemic to China, was collected from a stream in Hubei province, China. Unialgal cultivation, morphological observation, and phylogenetic analyses of small subunit ribosomal DNA and RuBisCO large subunit sequences were performed. When cultured on agar medium, the alga was irregularly filamentous, similar to marine species of Acrochaete. Aplanospores were observed on solid medium. A vesicular‐like thallus without rhizoids developed in liquid medium, similar to specimen development in natural habitats. Molecular phylogenetic analyses revealed that Jaoa was closely related to the marine genera Acrochaete Pringsheim and Ulvella Crouan & Crouan. The results suggested the genus Jaoa is a member of the family Ulvellaceae (Ulvophyceae), which contains mostly marine algae. The family name Jaoaceae should be abandoned. We speculate that Jaoa may have evolved from a marine Ulvellaceae ancestor.