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141.
试验旨在研究伪狂犬病病毒(PRV)在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞(3D4/21)p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。 相似文献
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Mingli LIN Mingming LIU Francesco CARUSO Massimiliano ROSSO Xiaoming TANG Lijun DONG Wenzhi LIN Anna BORRONI Alessandro BOCCONCELLI Liang DAI Songhai LI 《Integrative zoology》2021,16(4):440-450
A pioneering boat-based survey was conducted in 2019, to gather baseline information regarding the presence, composition, relative abundance, and spatial distribution of deep-diving and off-shore cetaceans in the northern South China Sea (SCS). A total of 27 sightings comprising at least 8 cetacean species were recorded during the 13-day survey, including 5 deep-diving species (i.e. Risso's dolphin [Grampus griseus], short-finned pilot whale [Globicephala macrorhynchus], sperm whale [Physeter macrocephalus], Cuvier's beaked whale [Ziphius cavirostris], and an unidentified beaked whale [either the ginkgo-toothed beaked whale, Mesoplodon ginkgodens, or Deraniyagala's beaked whale, Mesoplodon hotaula]), as well as 3 off-shore dolphins (i.e. pantropical spotted dolphin [Stenella attenuate], striped dolphin [Stenella coeruleoalba], and Fraser's dolphin [Lagenodelphis hosei]). With the exception of pantropical spotted dolphins, all other species were sighted and recorded at sea in the northern SCS for the first time. The pantropical spotted dolphin was the most frequently sighted species, comprising 30% of the total sightings. Deep-diving cetaceans were mainly sighted in the northern Xisha Archipelago, whereas off-shore dolphins were distributed across the survey area. The pantropical spotted dolphin was observed in aggregations of more than 100 individuals and nearly all encountered species included calves; these findings suggested that the survey area functions as an important feeding and calving ground for various cetacean species. This pioneering survey provides fundamental information regarding cetacean fauna in the northern SCS and highlights the need to strengthen research and conservation efforts concerning these species. 相似文献
144.
Yanfen Ma Ying Feng Liwen Song Muyang Li Hongyu Dai Hua Bao Guijie Zhang Lei Zhao Chunhua Zhang Jing Yi Yusheng Liang 《动物营养(英文)》2021,7(1):206
Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation. 相似文献
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采用湿法工艺生产玉米变性淀粉时,湿淀粉通常采用气流干燥,根据用户需要,设计了一套负压气流干燥设备来满足玉米变性淀粉的干燥。文章介绍设备主要结构、工作原理,并对干燥必需的能耗、所需的风量和热量、干燥管的长度、引风机功率等参数进行计算。设备经过实际生产使用,能够满足玉米变性淀粉的干燥要求。 相似文献
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通过对青海省高寒牧区常见的9种多年生牧草单播2年后耕层0~15cm土壤理化(pH、容重(BD)、有机碳(SOC)、全氮(TN)、无机碳(C)及微生物学性质(微生物生物量碳(Cmic)、氮(Nmic)和群落代谢功能)等指标的测定分析,结果表明,研究区域只有在种植披碱草2年后土壤有机碳含量有所增加,说明与其他草种相比,种植披碱草利于有机质的积累;试验在每年施肥1次的情况下,土壤氮含量仍然偏低,说明此区氮素被过度利用,处于缺乏水平,因此每年增施氮肥数量、频率以及时间上应加强管理。通过对不同牧草种植区土壤各因子的聚类分析,发现贫花鹅观草、无芒雀麦、紫野麦草和扁穗冰草之间相似度较高,表明其对土壤养分及微生物群落功能的影响较为接近,故在大面积种植的时候可根据牧草地上生物量/质量的高低进行选择性播种。从土壤质量方向考虑,种植杂花苜蓿、红豆草和西北羊茅不利于土地的改良。 相似文献