A rapid real-time recombinase polymerase amplification assay for diagnosis of acute hepatopancreatic necrosis disease in shrimp

Acute hepatopancreatic necrosis disease(AHPND)is an emerging disease in the shrimp farming industry with a high mortality rate that causes serious economic loss...     

核因子E2相关因子2在运动改善非酒精性脂肪性肝病中的作用

核因子E2相关因子2 (nuclear factor erythroid 2-related factor 2,Nrf2)信号通路在维持心血管疾病、神经系统退行性疾病以及慢性代谢性疾病中的细胞稳态方面起关键作用。研究表明,以氧化应激、炎症和线粒体功能失调为特征的慢性疾病可通过增加Nrf2表达来恢复机体氧化还原状态,治疗或预防疾病。非酒精性脂肪性肝病（nonalcoholic fatty liver disease,NAFLD）是一种由除酒精以外的其他多种因素导致的以肝脏脂肪变性为特征的慢性代谢性肝脏疾病,其患病率近年来在全球范围内逐渐增加。运动是防治NAFLD的有效手段,可通过运动方式、运动强度、运动环境和运动疲劳等因素影响Nrf2信号通路。本文通过阐述Nrf2信号通路的激活、其调控抗氧化的相关机制以及运动对Nrf2信号通路的影响,以NAFLD的发病机制为基础,探讨运动、Nrf2和NAFLD之间的关系,综述Nrf2在运动改善NAFLD中的作用及相关机制。为运动改善NAFLD的分子机制研究提供理论参考依据。     

猪圆环病毒2型流行株的分离及其感染性克隆的构建

【目的】通过分离一株猪圆环病毒2型(PCV2)流行毒株,并构建其感染性克隆,为研究PCV2基因功能提供操作平台。【方法】通过PCR方法,从疑似患断奶仔猪多系统衰竭综合症(PMWS)的仔猪淋巴结中鉴定为猪圆环病毒(Porcine circovirus,PCV)2型阳性。把阳性病料接种PK-15细胞传代培养,在培养物中扩增出PCV2的全基因序列。对扩增出的全序列进行序列测定,并与GenBank中公布的5株广东PCV2分离株(GD-pz、GD-gj、GD-jm、GD-ss和GD-sz)进行同源性分析。通过EcoRⅠ和SalⅠ将PCV2全基因组序列克隆进pUC18载体中,获得含PCV2 GD-zq株全基因组单拷贝的重组质粒pPCV2-GD-zq,再通过SalⅠ和HindⅢ把另一个全长拷贝克隆进pPCV2-GD-zq质粒中,使PCV2 GD-zq株基因组DNA以头尾相接的双重复方式克隆进pUC18载体中,获得重组质粒pPCV2-2GD-zq。将pPCV2-2GD-zq DNA纯化和定量后转染PK-15细胞,拯救PCV2 GD-zq病毒。【结果】从PMWS感染的猪淋巴结中分离到了一株PCV2,命名为GD-zq株;序列分析结果显示,GD-zq株全基因组为1 767 bp,与GenBank中公布的5株广东PCV2分离株ORF1核苷酸一致性为97.1%-99.7%,编码氨基酸一致性为98.7%-100%;ORF2核苷酸一致性为93.2%-99.6%,编码氨基酸一致性为92.3%-99.1%;全基因一致性为96.0%-99.6%。pPCV2-2GD-zq质粒转染PK-15细胞后,其通过间接免疫荧光实验(IFA)能从转染细胞及其传代细胞中,检测到拯救出的病毒。【结论】分离了一株PCV2广东株GD-zq,成功构建了PCV2 GD-zq株的感染性克隆。     

城市景观生态风险评估框架与实践——以北京天坛地区为例

当前,城市景观生态风险研究缺少科学合理、方便实用的评估框架。作者基于景观生态风险评估基本范式,明确了城市景观生态服务价值的测算方法,分析了引起生态损害的自然因素和人类活动因素,形成了城市景观生态风险评估的技术框架和参数体系;继而以北京天坛地区为研究区,开展了典型城市景观生态风险的定量评估。结果表明：（1）天坛地区景观生态价值总量约为2.41亿元。区域的历史文化价值最高,教育和美学景观价值紧随其后。（2）城市景观生态受损概率呈现"北高南低"的空间分布格局。生态受损概率的高值区面积占整个区域总面积的22.2%,主要分布在珠市口、磁器口和崇文门附近区域。（3）城市景观生态风险呈现"北低南高"的空间分布格局。高风险区主要分布在天坛公园内的文物建筑周边。本研究提供了一个可参考的城市景观生态风险评估应用框架,对生态风险评估中的不确定性进行了讨论,研究针对天坛案例区的具体结论有助于城市管理者避免潜在的风险。     

Regulation of progesterone during follicular development by FSH and LH in sheep

Progesterone (P4) can participate in the development of female mammalian antral follicles through nuclear receptor (PGR). In this experiment, the differences of P4 synthesis and PGR expression in different developmental stages of sheep antral follicles (large > 5mm, medium 2-5mm, small < 2mm) were detected by enzyme-linked immunosorbent assay, immunohistochemistry, qRT-PCR and Western blotting. Secondly, sheep follicular granulosa cells were cultured in vitro. The effects of different concentrations of FSH and LH on P4 synthesis and PGR expression were studied. The results showed that acute steroid regulatory protein (StAR), cholesterol side chain lyase (P450scc) and 3β Hydroxysteroid dehydrogenase (3β-HSD) and PGR were expressed in antral follicles, and with the development of antral follicles in sheep, StAR, P450scc and the expression of 3β-HSD and PGR increased significantly. In vitro experiments showed that FSH and LH alone or together treatment could regulate P4 secretion and PGR expression in sheep follicular granulosa cells to varying degrees, hint P4 and PGR by FSH and LH, and LH was the main factor. Our results supplement the effects of FSH and LH on the regulation of P4 synthesis during follicular development, which provides new data for further study of steroid synthesis and function in follicular development.     

南方型杨树人工林土壤呼吸及其组分分析

采用开沟隔离法,利用LI-8100型土壤呼吸测定系统,对15年生的南方型杨树(Populus deltoides)人工林土壤呼吸进行了研究,并试图区分根系呼吸和土壤微生物呼吸。结果表明,开沟隔离处理后的10个月内,由于土壤中被截断根系具有自养呼吸和分解作用,土壤呼吸中的根系呼吸与微生物呼吸尚难以区分。尽管如此,研究表明15年生杨树人工林的土壤总呼吸通量为9.74 tC.hm-.2a-1,其中,枯枝落叶等土壤表层凋落物分解所释放的碳通量是2.63 tC.hm-.2a-1,占总量的27.0%;林木根系呼吸与土壤微生物呼吸通量的和为7.11 tC.hm-.2a-1,占总量的73.0%。土壤各组分呼吸速率与10 cm深处的土壤温度之间存在着显著的指数函数关系。不同直径的杨树根系被截断后的活力变化有所不同,根系越粗,存活时间越长。     

辽河口滨海湿地CH4排放特征及其影响因素

采用静态箱-气相色谱法,于2016年6—11月连续观测辽河口芦苇湿地、翅碱蓬湿地和裸滩湿地的CH_4排放速率,同时测定温度、氧化还原电位(Eh)、pH值和电导率(EC)等相关环境因子的动态变化。结果表明,3种类型湿地的CH_4排放具有明显的季节变化特征,均呈先上升后下降趋势。芦苇湿地、翅碱蓬湿地(涨潮前)和裸滩湿地(涨潮前)CH_4排放通量变化范围分别为0.447—10.40、0.045—0.509 mg m~(-2) h~(-1)和0.016—0.593 mg m~(-2) h~(-1),观测期内排放通量均值相应为(3.699±3.679)、(0.165±0.156) mg m~(-2) h~(-1)和(0.198±0.191) mg m~(-2) h~(-1),不同类型湿地之间差异显著(P0.01),芦苇湿地裸滩湿地(涨潮前)翅碱蓬湿地(涨潮前)。涨潮过程中,翅碱蓬湿地和裸滩湿地的排放速率分别变化在0.009—0.353 mg m~(-2) h~(-1)和0.018—0.335 mg m~(-2) h~(-1),观测期间其排放速率均值分别为(0.119±0.132) mg m~(-2) h~(-1)和(0.131±0.103) mg m~(-2) h~(-1),明显低于涨潮前(P0.01)。不同湿地类型间CH_4排放通量与电导率(EC)呈显著负相关(P0.01)。研究结果表明,潮汐和电导率均为影响辽河口不同类型湿地中CH_4排放的关键因子。     

Glycosylated modification of MUC1 maybe a new target to promote drug sensitivity and efficacy for breast cancer chemotherapy

Breast cancer, the most common cancer in women, usually exhibits intrinsic insensitivity to drugs, even without drug resistance. MUC1 is a highly glycosylated transmembrane protein, overexpressed in breast cancer, contributing to tumorigenesis and worse prognosis. However, the molecular mechanism between MUC1 and drug sensitivity still remains unclear. Here, natural flavonoid apigenin was used as objective due to the antitumor activity and wide availability. MUC1 knockout (KO) markedly sensitized breast cancer cells to apigenin cytotoxicity in vitro and in vivo. Both genetical and pharmacological inhibition significantly enhanced the chemosensitivity to apigenin and clinical drugs whereas MUC1 overexpression conversely aggravated such drug resistance. Constitutively re-expressing wild type MUC1 in KO cells restored the drug resistance; however, the transmembrane domain deletant could not rescue the phenotype. Notably, further investigation discovered that membrane-dependent drug resistance relied on the extracellular glycosylated modification since removing O-glycosylation via inhibitor, enzyme digestion, or GCNT3 (MUC1 related O-glycosyltransferase) knockout markedly reinvigorated the chemosensitivity in WT cells, but had no effect on KO cells. Conversely, inserting O-glycosylated sites to MUC1-N increased the drug tolerance whereas the O-glycosylated deletant (Ser/Thr to Ala) maintained high susceptibility to drugs. Importantly, the intracellular concentration of apigenin measured by UPLC and fluorescence distribution firmly revealed the increased drug permeation in MUC1 KO and BAG-pretreated cells. Multiple clinical chemotherapeutics with small molecular were tested and obtained the similar conclusion. Our findings uncover a critical role of the extracellular O-glycosylation of MUC1-N in weakening drug sensitivity through acting as a barrier, highlighting a new perspective that targeting MUC1 O-glycosylation has great potential to promote drug sensitivity and efficacy.Subject terms: Breast cancer     

Electron acceptors for anaerobic oxidation of methane drive microbial community structure and diversity in mud volcanoes

Mud volcanoes (MVs) emit globally significant quantities of methane into the atmosphere, however, methane cycling in such environments is not yet fully understood, as the roles of microbes and their associated biogeochemical processes have been largely overlooked. Here, we used data from high‐throughput sequencing of microbial 16S rRNA gene amplicons from six MVs in the Junggar Basin in northwest China to quantify patterns of diversity and characterize the community structure of archaea and bacteria. We found anaerobic methanotrophs and diverse sulfate‐ and iron‐reducing microbes in all of the samples, and the diversity of both archaeal and bacterial communities was strongly linked to the concentrations of sulfate, iron and nitrate, which could act as electron acceptors in anaerobic oxidation of methane (AOM). The impacts of sulfate/iron/nitrate on AOM in the MVs were verified by microcosm experiments. Further, two representative MVs were selected to explore the microbial interactions based on phylogenetic molecular ecological networks. The sites showed distinct network structures, key species and microbial interactions, with more complex and numerous linkages between methane‐cycling microbes and their partners being observed in the iron/sulfate‐rich MV. These findings suggest that electron acceptors are important factors driving the structure of microbial communities in these methane‐rich environments.