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Metabolomics and water migration analysis provides valuable insights into nutrient generation in Tartary buckwheat (Fagopyrum tataricum) seed germination 代谢组学和水分迁移分析为苦麦种子发芽过程中的营养生成提供了有价值的见解
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-09-19 DOI: 10.1080/09540105.2022.2117797
Bai Yongliang, Xin Meiguo, Lin Roumin, He Weijun, He Shuyan, Z. Rong, Guo Yiping
ABSTRACT Seed germination of Tartary buckwheat is important for its biological generation of various nutrients. To investigate the nutrient differences during seed germination, the metabolic profiling and moisture status of Tartary buckwheat seed germination were quantitatively measured by ultra-high performance liquid chromatography mass spectrometry (UHPLC-MS) and NMR, respectively. The results showed water was predominantly in free status during the three stages of seed germination divided. Statistical analysis (PCA, OPLS-DA, KEGG) of 53 metabolites at different germination stages revealed the key metabolites of linoleic, flavonoid and phenylalanine biosynthesis during the seed germination. Furthermore, flavonoids biosynthesis was identified as the pathway with the largest difference significance at the second and third stages, while linoleic and phenylalanine metabolisms were screened out as the major pathways with most impact factors in the first and second and the third stage, respectively. The analysis provided valuable insights into the nutrient generation during Tartary buckwheat seeds germination.
荞麦种子的发芽对其生物生成各种营养物质具有重要意义。为了研究种子发芽过程中的营养差异,分别用超高效液相色谱-质谱(UHPLC-MS)和核磁共振(NMR)对苦麦种子发芽过程的代谢谱和水分状况进行了定量测定。结果表明,在种子萌发的三个阶段中,水分主要处于游离状态。对53种不同发芽阶段代谢产物的统计分析(PCA、OPLS-DA、KEGG)揭示了种子发芽过程中亚油酸、类黄酮和苯丙氨酸生物合成的关键代谢产物。此外,类黄酮生物合成在第二阶段和第三阶段被确定为差异显著性最大的途径,而亚油酸和苯丙氨酸代谢分别在第一阶段和第二阶段及第三阶段被筛选为影响因素最多的主要途径。该分析为苦荞麦种子发芽过程中的营养生成提供了有价值的见解。
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引用次数: 0
Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for assessing okadaic acid and its derivatives in shellfish 快速酶联免疫吸附法和胶体金免疫测定法测定贝类中冈田酸及其衍生物
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-31 DOI: 10.1080/09540105.2022.2115466
Ruirui Wang, Hong-bo Qi, Miao Liang, Guoxiang Liao, Fan Yang
ABSTRACT Okadaic acid (OA) and the analogs of dinophysistoxin (DTX) are important members of diarrhetic shellfish poisoning (DSP) toxins. In this study, five OA-specific mAbs (monoclonal antibodies) were developed from five stable cells of hybridoma. The anti-OA mAb-2D7 showed high sensitivity to OA, the IC50 of the antibody was 0.24 ng/mL, and its cross-reactivity was 91.6% with DTX-1 and 110.5% with DTX-2., In dcELISA, the IC50 was set at 0.182 ng/mL, and the detection limit was set at 0.023 ng/mL by using the anti-OA mAb-2D7. The level of OA recovered from spiked mussel samples was 2–50 ng/g, ranging from 97.6 ± 7.2% to 106.4 ± 9.8%. In contrast, the immunostrip assay with a limit of 5 ng/mL, conducted for detecting OA, was completed in 10 min. The mAb-based dcELISA and immunostrip assay developed were precise and sensitive enough to quickly assess OA, DTX-1, and DTX-2 in shellfish specimens.
冈田酸(OA)及其类似物是腹泻性贝类中毒(DSP)毒素的重要组成部分。在本研究中,从五种稳定的杂交瘤细胞中开发了五种OA特异性单克隆抗体。抗OA mAb-2D7对OA具有较高的敏感性,其IC50为0.24ng/mL,与DTX-1和DTX-2的交叉反应性分别为91.6%和110.5%。从加标贻贝样品中回收的OA水平为2–50纳克/克,范围为97.6 ± 7.2%至106.4 ± 9.8%。相比之下,免疫跳闸试验的限值为5 用于检测OA的ng/mL在10分钟内完成。开发的基于mAb的dcELISA和免疫跳闸测定足够精确和灵敏,可以快速评估贝类标本中的OA、DTX-1和DTX-2。
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引用次数: 4
Curcumin inhibits invasion and metastasis of human hepatoma cells through Bclaf1-mediated Wnt/β-catenin signalling 姜黄素通过bclaf1介导的Wnt/β-catenin信号传导抑制人肝癌细胞的侵袭和转移
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-28 DOI: 10.1080/09540105.2022.2113864
Zhongwei Zhao, Jielin Su, Jiaqi Zhao, Jiaxin Chen, Xinmu Cui, Manqing Sun, Xuewu Zhang
ABSTRACT Curcumin, a kind of natural compound extracted from the rhizome of Zingiberaceae such as turmeric, has many pharmacological effects such as anti-cancer effects. This study investigated the effect of curcumin on the invasion and metastasis of hepatocellular carcinoma (HCC) cell lines HepG2 and SK-Hep-1 through the Wnt/β-catenin signalling pathway and the regulatory mechanism of Bcl-2-associated transcription factor 1 (Bclaf1). Curcumin significantly inhibited the migration and invasion of HepG2 and SK-Hep-1 cells and inhibited the Wnt/β-catenin signalling pathway and reduced Bclaf1 expression in human hepatoma cells. In nude mice, intraperitoneal injection of curcumin significantly inhibited the growth of subcutaneously transplanted tumours and reduced lung metastasis of the tumour cells, downregulated the expression of Bclaf1, and inhibited the Wnt/β-catenin pathway. This study suggests that curcumin is a novel candidate drug to prevent cancer metastasis and that Bclaf1 is a new gene target related to the proliferation, invasion, and metastasis of hepatocellular carcinoma.
摘要姜黄素是从姜科姜根中提取的一种天然化合物,具有抗癌等药理作用。本研究探讨了姜黄素通过Wnt/β-catenin信号通路对肝癌细胞系HepG2和SK-Hep-1侵袭转移的影响,以及Bcl-2相关转录因子1(Bclaf1)的调控机制。姜黄素显著抑制HepG2和SK-Hep-1细胞的迁移和侵袭,抑制Wnt/β-catenin信号通路,降低Bclaf1在人肝癌细胞中的表达。在裸鼠中,腹腔注射姜黄素显著抑制皮下移植肿瘤的生长,减少肿瘤细胞的肺转移,下调Bclaf1的表达,并抑制Wnt/β-catenin通路。本研究表明,姜黄素是一种新的预防癌症转移的候选药物,Bclaf1是一种与肝细胞癌增殖、侵袭和转移相关的新基因靶点。
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引用次数: 1
Identification and characterization of MicroRNAs in pig liver after the LPS challenge using RNA-seq 利用RNA-seq技术鉴定和表征LPS攻毒后猪肝中的microrna
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-22 DOI: 10.1080/09540105.2022.2109602
Yang Yang, Qingqing Lv, Xingfa Huang, Jiajun Fan, Pei Li, Huijuan Zhu, P. Kang, Yulan Liu
ABSTRACT This study was conducted to test the expression profiles of miRNAs in the liver in piglets after the LPS challenge using sequencing technology. Twelve barrows were assigned to two groups, including the saline group and the LPS group. Six small RNA (sRNA) libraries were constructed, and novel miRNAs were identified using mirevo and mirdeep2 software and validated by quantitative real-time PCR (qRT-PCR). The results showed the mRNA expression of IL-1β, IL-6 and TNF-α in the liver increased after the LPS challenge (P < .05). A total of 29 differentially expressed miRNAs were identified in the liver after the LPS challenge. And 11 miRNAs were validated by qRT-PCR. In addition, the results of statistics of pathway enrichment showed that these miRNAs could be related to lipid metabolism response. This study provides the first miRNA expression profiles of LPS-mediated changes in the liver, which might provide potential insights into miRNAs involved in regulating lipid metabolism in pigs challenged by LPS.
本研究采用测序技术检测LPS攻毒后仔猪肝脏中mirna的表达谱。将12只雄鼠分为生理盐水组和LPS组。构建6个小RNA (sRNA)文库,利用mirevo和mirdeep2软件鉴定新的mirna,并采用实时荧光定量PCR (qRT-PCR)进行验证。结果显示,LPS刺激后肝脏中IL-1β、IL-6和TNF-α mRNA表达升高(P < 0.05)。LPS刺激后,肝脏中共鉴定出29种差异表达的mirna。通过qRT-PCR对11个mirna进行了验证。此外,途径富集的统计结果表明,这些mirna可能与脂质代谢反应有关。本研究首次提供了LPS介导的肝脏变化的miRNA表达谱,这可能为LPS刺激下参与调节猪脂质代谢的miRNA提供潜在的见解。
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引用次数: 1
Solanum nigrum induces macrophage activation through TLR4-mediated activation of JNK and macrophage autophagy through TLR4-mediated activation of p38 and JNK 龙葵通过TLR4介导的JNK活化诱导巨噬细胞活化,并通过TLR4中介导的p38和JNK活化介导巨噬细胞自噬
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-16 DOI: 10.1080/09540105.2022.2113507
Julia Yu, Min Yeong Choi, So-Jung Park, Na Gyeong Geum, Jae Won Lee, G. Park, H. Eo, J. Jeong
ABSTRACT In this study, we investigated the effect of Solanum nigrum aerial parts (SNAP) on macrophage activation and macrophage autophagy in RAW264.7 cells. SNAP increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. TLR4 inhibition blocked SNAP-mediated production of immunostimulatory factors. In addition, the JNK inhibition reduced the SNAP-mediated production of immunostimulatory factors, and the SNAP-mediated JNK activation was blocked by the TLR4 inhibition. SNAP activated macrophage autophagy. TLR4 inhibition blocked SNAP-mediated macrophage autophagy and inhibition of p38 and JNK attenuated SNAP-mediated macrophage autophagy. These findings indicate that SNAP may induce TLR4/JNK-mediated macrophage activation and TLR4/p38 and JNK-mediated macrophage autophagy.
摘要在本研究中,我们研究了龙葵地上部分(SNAP)对RAW264.7细胞巨噬细胞活化和巨噬细胞自噬的影响。SNAP增加了RAW264.7细胞中免疫刺激因子的产生和吞噬作用。TLR4抑制阻断SNAP介导的免疫刺激因子的产生。此外,JNK抑制降低了SNAP介导的免疫刺激因子的产生,并且SNAP介介导的JNK激活被TLR4抑制所阻断。SNAP激活巨噬细胞自噬。TLR4的抑制阻断了SNAP介导的巨噬细胞自噬,p38和JNK的抑制减弱了SNAP中介导的巨噬细胞自噬。这些发现表明SNAP可能诱导TLR4/JNK介导的巨噬细胞活化以及TLR4/p38和JNK介导的细胞自噬。
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引用次数: 0
Immune-enhancing effects of Hibiscus syriacus roots in RAW264.7 macrcophages 木槿根对RAW264.7巨噬细胞的免疫增强作用
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-16 DOI: 10.1080/09540105.2022.2107621
H. Eo, Yunmi Park, H. Kwon, G. Park
ABSTRACT In this study, we evaluated whether extracts of the roots of Hibiscus syriacus (HSR) exert immune activation activities and elucidated its potential mechanism in macrophages. The HSR dose-dependently increased the production of immunomodulators such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor (TNF-α) activated phagocytosis in macrophages. Inhibition of toll-like receptors 4 (TLR4) reduced the production of immunomodulators induced by HSR. Inhibition of mitogen-activated protein kinase (MAPKs), nuclear factor-κB (NF-κB) and phosphoinositide-3 kinase (PI3K) signaling attenuated the production of immunomodulators induced by HSR. Based on the results of this study, HSR was thought to activate macrophages through the production of immunomodulators and phagocytosis activation through TLR4-dependent MAPKs, NF-κB and PI3K signaling pathways. Therefore, it is thought that the HSR has the potential to be used as agents for enhancing immunity.
摘要本研究研究了木槿根提取物(HSR)在巨噬细胞中是否具有免疫激活活性,并阐明了其潜在机制。高铁剂量依赖性地增加免疫调节剂如一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)、环氧化酶-2 (COX-2)、白细胞介素-1β (IL-1β)和肿瘤坏死因子(TNF-α)在巨噬细胞中激活的吞噬作用。抑制toll样受体4 (TLR4)可减少HSR诱导的免疫调节剂的产生。抑制丝裂原活化蛋白激酶(MAPKs)、核因子-κB (NF-κB)和磷酸肌醇激酶(PI3K)信号可减弱HSR诱导的免疫调节剂的产生。基于本研究的结果,我们认为HSR通过tlr4依赖性的MAPKs、NF-κB和PI3K信号通路,通过产生免疫调节剂和吞噬激活来激活巨噬细胞。因此,人们认为高铁具有作为增强免疫力的药剂的潜力。
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引用次数: 0
Design and synthesis of geosmin derivatives using organic synthesis strategies and application in antibody production 利用有机合成策略设计、合成geosmin衍生物及其在抗体生产中的应用
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-11 DOI: 10.1080/09540105.2022.2107620
Penghui Zhou, Ying Liu, Ting Zhou, Hanwen Liu, Jianguo Li, Anping Deng
ABSTRACT To develop a sensitive and specific ELISA for geosmin (GSM), the research of this study is focused on the design and synthesis of several GSM derivatives, which maintain the whole GSM structure as much as possible and contain a spacer arm with an active group at the end. To form the GSM backbone, tandem organic reaction strategies were used to replace the traditional Robinson cyclization reaction. Five GSM derivatives were synthesized and the formed GSM derivative-protein conjugates were used as the immunogens for the production of polyclonal antibodies against GSM, while four GSM derivatives were synthesized and the GSM derivative-protein conjugates were used as coating antigens for establishing ELISA. The relationship between the structures of GSM derivatives and the antibody properties was explored. Under optimal conditions, the LOD of the ELISA for GSM was found to be 0.16 ng mL−1, and the antiserum was able to specifically recognize the GSM backbone.
摘要为了开发一种灵敏、特异的geosmin(GSM)ELISA,本研究的重点是设计和合成几种GSM衍生物,这些衍生物尽可能保持整个GSM结构,并包含末端带有活性基团的间隔臂。为了形成GSM主链,使用串联有机反应策略来取代传统的Robinson环化反应。合成了5种GSM衍生物,并将形成的GSM衍生物蛋白偶联物用作制备抗GSM多克隆抗体的免疫原,同时合成了4种GSM衍生物并将GSM衍生物蛋白缀合物用作建立ELISA的包被抗原。探讨了GSM衍生物的结构与抗体性质之间的关系。在最佳条件下,ELISA检测GSM的LOD为0.16 ng mL−1,并且该抗血清能够特异性识别GSM主链。
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引用次数: 0
Taraxasterol repairs UVB-induced skin barrier injury through MAPK/NF-κB signaling pathways 蒲公英醇通过MAPK/NF-κB信号通路修复uvb诱导的皮肤屏障损伤
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-04 DOI: 10.1080/09540105.2022.2107619
Yuzhi Zhang, Hao Fu, Yongtao Zhang, Dongdong Wang, Dan Zhao, Jiachan Zhang, Meng Li, Changtao Wang
ABSTRACT Taraxasterol (TAL) is a pentacyclic triterpenoid compound, which has anti-inflammatory effect. Cytotoxicity assay was used to determine the optimal concentration of TAL and positive control dipotassium glycyrrhizinate (DG), and the optimal dose of UVB. Experimental data indicate that TAL has scavenging activity against UVB radiation-induced intracellular reactive oxygen species (ROS) compared to UVB controls. The contents of skin barrier-related factors in the groups were detected by Enzyme-linked immunosorbent assay (ELISA), then ELISA and quantitative polymerase chain reaction (qPCR) were used to detect changes in the inflammatory factors, apoptosis factors, and gene levels in the groups. Therefore, TAL stabilised the levels of inflammation, apoptosis, and skin barrier-related factors by regulating Mitogen-activated protein kinases/nuclear factor-k-gene binding (MAPK/NF-κB) signalling pathways, such as jun-amino-terminal kinase (JNK), p38, extracellular signal-regulated kinase (ERK), and NF-κB. These results suggest that TAL repairs UVB-induced skin barrier damage by scavenging reactive oxygen species and regulating the MAPK/NF-κB signalling pathway.
塔拉沙甾醇(TAL)是一种具有抗炎作用的五环三萜类化合物。采用细胞毒性试验确定TAL和阳性对照甘草酸二钾(DG)的最佳浓度以及UVB的最佳剂量。实验数据表明,与UVB对照相比,TAL对UVB辐射诱导的细胞内活性氧(ROS)具有清除活性。采用酶联免疫吸附试验(ELISA)检测各组皮肤屏障相关因子的含量,然后采用ELISA和定量聚合酶链式反应(qPCR)检测各组炎症因子、细胞凋亡因子和基因水平的变化。因此,TAL通过调节丝裂原活化蛋白激酶/核因子-激酶结合(MAPK/NF-κB)信号通路,如jun氨基末端激酶(JNK)、p38、细胞外信号调节激酶(ERK)和NF-κ。这些结果表明,TAL通过清除活性氧和调节MAPK/NF-κB信号通路来修复UVB诱导的皮肤屏障损伤。
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引用次数: 1
Sensitisation to molecular components of mites in atopic dermatitis patients 特应性皮炎患者对螨虫分子成分的致敏
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-08-04 DOI: 10.1080/09540105.2022.2100323
J. Čelakovská, E. Čermáková, R. Vankova, P. Boudková, C. Andrys, J. Krejsek
ABSTRACT Background and aim: To analyse the sensitisation profile to molecular components of mites allergens with the use of ALEX2 Allergy Explorer test in Atopic dermatitis (AD) patients. Method: The complete dermatological and allergological examination including the examination of the sensitisation to molecular components with ALEX2 Allergy Explorer testing was performed. For the statistical analysis, we used Fisher's Exact. Results and conclusion: The sensitisation to mites increases with AD severity. The central role in atopic march may play the molecular component Der f 2 and Der p 23. The results of our study confirm the important role of Der p 5 and Der p 7 in subgroup of patients suffering from bronchial asthma. The specific immunotherapy should focus on the components of NPC2 family and Der p 23 in AD patients. Our results may be helpful in planning hyposensitisation, treatment and immunotherapy.
摘要背景和目的:应用ALEX2过敏探索者试验分析特应性皮炎(AD)患者对螨类过敏原分子成分的致敏情况。方法:进行完整的皮肤科和变态反应学检查,包括用ALEX2 Allergy Explorer测试对分子成分的敏感性。对于统计分析,我们使用了Fisher精确。结果和结论:对螨虫的敏感性随着AD的严重程度而增加。特应性进行曲的核心作用可能是分子组分Der f2和Der p23。我们的研究结果证实了Der p 5和Der p 7在支气管哮喘患者亚组中的重要作用。特异性免疫治疗应关注AD患者的NPC2家族和Der p 23的成分。我们的研究结果可能有助于规划减敏、治疗和免疫治疗。
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引用次数: 0
A highly salt-tolerant monoclonal antibody-based enzyme-linked immunosorbent assay for the rapid detection of phenylethanolamine A in urine 基于高耐盐单克隆抗体的酶联免疫吸附法快速检测尿液中苯乙醇胺A
IF 3 3区 农林科学 Q3 CHEMISTRY, APPLIED Pub Date : 2022-07-29 DOI: 10.1080/09540105.2022.2084043
Minggang Liu, Yuchen Bai, Leina Dou, Yihui Kong, Zhanhui Wang, K. Wen, Jianzhong Shen
ABSTRACT Phenylethanolamine A (PEAA), as a typical β2-adrenoceptor agonist (β-AA), was widely illegally used in feed to improve the lean meat ratio. The detection performance of immunoassay, which is one of the effective methods to monitor PEAA residues in animal urine, has been hindered by high concentration of salt. Herein, we produced one highly salt-tolerant monoclonal antibody (mAb) 3E2 by hybridoma technology. Homologous modelling and molecular docking were used to analyse the antibody salt tolerance mechanism. Results show that the tight hydrophobic binding cavity is the key to high tolerance. Then the mAb 3E2 was used for specific detecting PEAA by enzyme-linked immunosorbent assay (ELISA) with the IC50 value of 0.36 ng mL−1and the LOD value of 0.065 µg L−1. Benefiting from the high salt tolerance of antibodies, swine urine and sheep urine spiked with PEAA can be detected directly by ELISA, and the acceptable recovery rates of 80.1–108.8% were obtained.
摘要苯乙醇胺A (PEAA)作为一种典型的β2-肾上腺素能受体激动剂(β-AA),被广泛非法用于饲料中以提高瘦肉比。免疫分析法是动物尿液中PEAA残留的有效检测方法之一,但高浓度盐的存在影响了其检测效果。本文利用杂杂瘤技术制备了一种高耐盐单克隆抗体(mAb) 3E2。采用同源建模和分子对接分析了抗体耐盐机理。结果表明,紧密的疏水结合腔是高耐受性的关键。采用酶联免疫吸附法(ELISA)特异性检测PEAA, IC50值为0.36 ng mL−1,LOD值为0.065µg L−1。利用PEAA抗体耐盐性强的特点,猪尿和羊尿中PEAA的可接受回收率为80.1 ~ 108.8%。
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引用次数: 3
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Food and Agricultural Immunology
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