JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2016
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IF 2.687
Bin Chen,Qing Ye,Kangping Zhou,Yefu Wang
A novel type of aptamer-functionalized immunoadsorbent was prepared and characterized to remove HCV particles, a promising option of extracorporeal immunoadsorption (ECI) therapy against HCV. Herein, we fabricated a HCV-specific immunoadsorbent where single-stranded DNA aptamers reported and studied previously, modified with amino group at the 5′ terminus, was immobilized covalently onto surfaces of carboxylated-derivative sepharose 4FF beads through N -hydroxysuccinimide (NHS) linkage. Then the adsorbents was evaluated and characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Subsequently, we also confirmed that proposed immunoadsorbents exhibited a favorable biocompatibility as well as specificity. In addition, time-dependent effects of the eradication capacity of aptamer functionalized sepharose beads against HCV was investigated. With the optimized time point, the decontamination performance of HCV particles was assessed by real-time quantitative PCR (qPCR) followed by nucleic acid-based hybridization (NAH), which shows sorbents with an aptamer density of 2 nmol ligand/ml resin could remove approximately 80% (i.e. 8.9 × 10 6 HCV particles/ml resin) of the HCV genotype 2a cultivated in vitro and 75% (vary with the intial concentration of HCV from about 7.5 × 10 4 –4.4 × 10 5 HCV particles/ml resin) of the HCV samples from human plasma samples. All these results indicated that the novel aptamer-based adsorbents could effectively remove HCV particles and likely serve as a novel therapy option or at least supplementary for the treatment regimen of HCV.
研究领域:
病毒学
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分子生物学
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治疗学
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产品关联
DOI
PubMed
高影响因子
AQUACULTURE
·
2022
·
IF 5.135
Jia Liu,Hui Du,Tao Liu,Cheng Chen,Ying Yan,Tianqiang Liu,Lei Liu,Erlong Wang
Hepatopancreas as the main organ detoxifying xenobiotics with higher detoxifying and antioxidant enzymes activities , could best reflected the status of antioxidant defence against virus infection in the organisms. Catalase ( CAT ) and glutathione S-transferase ( GST ) were two important enzymes associated with hepatopancreas antioxidation and detoxification. To obtain the accurate expression levels of CAT and GST gene which reflect the status of hepatopancreas antioxidation and detoxification, two or more reference genes expressed stably under different conditions should be selected for RT-qPCR analysis. In this study, eight candidate reference genes were investigated using RefFinder program to screen the most stable reference gene during virus (white spot syndrome virus, WSSV) infection and drug (matrine) treatments in Procambarus clarkii , the relative expression levels and the activities of CAT and GST in hepatopancreas were detected and compared to verify the validity of selected reference genes. The results showed that taken different tissues together, EIF and EF1 α were the most stable and unstable reference genes respectively in P. clarkii no matter under different WSSV infection periods (0, 24, 48, 72 h) or different matrine treatment methods according to RefFinder analysis. The optimal number of reference genes in hepatopancreas was two using geNorm analysis, i.e. EIF and TBP , which were top two most stable reference genes in hepatopancreas. Moreover, based on the comparison between the relative expression levels and the activities of CAT and GST, EIF and TBP as reference genes were proved to be necessary and practicable to evaluate hepatopancreas antioxidation during WSSV infection and matrine treatments. Besides, the optimal number of reference genes in other P. clarkii tissues was also two genes, i.e. EIF and TUB in intestine, EIF and EF1α in muscle, EIF and GAPDH in gills, EIF and TBP in brain , TBP and UB in gonads . Our study was helpful to acquire the accurate data of antioxidation and detoxification-related and immune-related gene expression during virus infection and drug treatments in crayfish, especially in hepatopancreas tissues
研究领域:
水产养殖
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分子生物学
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免疫学
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兽医学
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产品关联
DOI
高影响因子
LAB ON A CHIP
·
2021
·Vol. 21
·期 18
·3429-3437
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IF 6.799
Tengbao Xie,Ping Wang,Lei Wu,Bangyong Sun,Qiang Zhao,Gang Li
In this work, we present a simple and equipment-free system for discretizing samples into tens of thousands of discrete volumes in tens of seconds. Unlike conventional sample discretization systems that require bulky syringe pumps, pressure controllers, or vacuum equipment, our system requires only a sheet of water-soluble film, a hand-operated syringe, and a microfluidic device containing a high-density microchamber array. In this system, the water-soluble film seals the device inlet to form a closed channel–chamber system, while the syringe is used to create a vacuum in the closed system. Benefitting from the high negative pressure created by syringe-vacuum and the dissolution-triggered gating mechanism of the sealing water-soluble film, the aqueous sample loaded into the device inlet can be rapidly partitioned into tens of thousands of isolated chambers without the need for any expensive pumping systems. We demonstrated the utility of this system by exploiting it for digital PCR. We believe that this simple discretization system will find broad applications, such as in digital bioassays, single-cell analysis, and point-of-care diagnostics.
研究领域:
微流控技术
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生物技术
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即时诊断
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数字生物分析
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单细胞分析
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产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2023
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IF 7.3
Zhang Lishan,Sun Lina,Srinivasan Ramanathan,Lin Meizhen,Gong Lanqing,Lin Xiangmin
Bacterial AraC is a transcription factor family that initiates transcription by recruiting RNA polymerase to the promoter and directly regulating various bacterial phenotypes. It also directly regulates various bacterial phenotypes. However, how this transcription factor regulates bacterial virulence and affects host immunity is still largely unknown. In this study, deleting the orf02889 (AraC-like transcription factor) gene in virulent Aeromonas hydrophila LP-2 affected several important phenotypes, such as increasing biofilm formation and siderophore production abilities. Moreover, Δorf02889 also significantly decreased the virulence of A. hydrophila and has promising attenuated vaccine potential. To better understand the effects of orf02889 on biological functions, a data independent acquisition (DIA)-based quantitative proteomics method was performed to compare the differentially expressed proteins between Δorf02889 and the wild-type strain in extracellular fractions. The following bioinformatics analysis suggested that ORF02889 may regulate various metabolic pathways, such as quorum sensing and ATP binding cassette (ABC) transporter metabolism. Moreover, 10 selected genes from the top 10 decreasing abundances in proteomics data were deleted, and their virulence to zebrafish was evaluated, respectively. The results showed that ΔcorC, Δorf00906, and Δorf04042 significantly reduced bacterial virulence. Finally, the following chromatin immunoprecipitation and polymerase chain reaction (ChIP-PCR) assay validated that the promoter of corC was directly regulated by ORF02889. Overall, these results provide insight into the biological function of ORF02889 and demonstrate its inherent regulatory mechanism for the virulence of A. hydrophila.
研究领域:
微生物学
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蛋白质组学
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分子生物学
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传染病学
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产品关联
DOI
PubMed
高影响因子
POSTHARVEST BIOLOGY AND TECHNOLOGY
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2023
·
IF 7
Fan Yang,Shuqi Liu,Zhonghuan Tian,Yujie Du,Deyao Zhang,Chao-an Long
Edetate salts are a kind of food additive regarded as candidates for chemical fungicides because of their lower toxicity and higher safety for the environment and human health. The antifungal activities of edetate salts have been determined in some phytopathogens except Penicillium digitatum , which causes the most destructive fungal disease during the citrus postharvest period. In the present study, we investigated the molecular mechanisms of calcium disodium edetate (EDTA-Na 2 Ca) against P. digitatum in deepth. The results suggested that EDTA-Na 2 Ca inhibited the P. digitatum P44 strain in vitro at a minimum inhibitory/fungicidal concentration (MIC/MFC) of 50 mmol L −1 . Meanwhile, EDTA-Na 2 Ca also inhibited P. digitatum in vivo by enhancing the host defense system. The P44 strain treated with EDTA-Na2Ca displayed a delayed conidial germination rate, expanded hyphal tips, thickened septa and cell walls, and destroyed mitochondria. Transcriptome sequencing and qRT-PCR analyses indicated that EDTA-Na 2 Ca suppressed the P44 strain primarily by disturbing the carbohydrate metabolism pathway, especially the synthesis of pyruvate in glycolysis. In addition, the adverse effects, including those on mycelial growth, pyruvate synthesis and superoxide dismutase activity, of EDTA-Na 2 Ca could be reversed by exogenous Mn 2+ , implying that Mn 2+ chelation is an essential mode of action. In summary, the current work verifies that EDTA-Na 2 Ca is a potential antifungal drug that cures citrus green mold probably by chelating Mn 2+ and targeting pyruvate synthesis in P. digitatum .
研究领域:
植物病理学
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分子生物学
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食品科学
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环境科学
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DOI
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2021
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IF 4.36
Yuling Tong,Ruilin Wang,Xia Liu,Miao Tian,Yanling Wang,Yanfei Cui,Wenjun Zou,Yanling Zhao
Ethnopharmacological relevance Zuojin Pill (ZJP) is a classic prescription composed of Coptis chinensis and Tetradium ruticarpum (A.Juss.) T.G.Hartley, which is often used in the treatment of digestive system diseases . Aim of this study The purpose of this study was to explore the therapeutic effect and potential mechanism of ZJP on chronic atrophic gastritis (CAG) induced by MNNG . Materials and methods The GES-1 and rat model of CAG was established by MNNG. Detection of cell viability, morphological changes and proliferation of GES-1 by CCK-8 and high content screening (HCS) assay. G-17, IL-8 and TNF-α in rat serum were detected by ELISA kit. The expression of related mRNA and protein on TGF-β1/PI3K/Akt signal axis were detected by RT-PCR and Western blot . Results The results showed that ZJP could significantly improve the GES-1 damage induced by MNNG and improve the gastric histomorphology of CAG rats. The intervention of ZJP could significantly reduce the content of G-17 and inflammatory factors IL-8, TNF- α, IL-6 and IL-1β, inhibit the expression of TGF-β1, PI3K and their downstream signals p-Akt, p-mTOR, P70S6K, and promote the expression level of PTEN, LC3-II and Beclin-1. Conclusion ZJP has a good therapeutic effect on CAG induced by MNNG, which may be closely related to the inhibition of TGF-β1/PI3K/Akt signal pathway.
研究领域:
药理学
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民族药理学
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胃肠病学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
PHYTOMEDICINE
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2021
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IF 5.34
Ying-yuan Zhao,Xin-peng Yao,Ting-ying Jiao,Jia-ning Tian,Yue Gao,Shi-cheng Fan,Pan-pan Chen,Yi-ming Jiang,Yan-ying Zhou,Yi-xin Chen,Xiao Yang,Min Huang,Hui-chang Bi
Background Schisandrol B (SolB) is one of the bioactive components from a traditional Chinese medicine Schisandra chinensis or Schisandra sphenanthera . It has been demonstrated that SolB exerts hepatoprotective effects against drug-induced liver injury and promotes liver regeneration . It was found that SolB can induce hepatomegaly but the involved mechanisms remain unknown. Purpose This study aimed to explore the mechanisms involved in SolB-induced hepatomegaly. Methods Male C57BL/6 mice were injected intraperitoneally with SolB (100 mg/kg) for 5 days. Serum and liver samples were collected for biochemical and histological analyses. The mechanisms of SolB were investigated by qRT-PCR and western blot analyses, luciferase reporter gene assays and immunofluorescence. Results SolB significantly increased hepatocyte size and proliferation, and then promoted liver enlargement without liver injury and inflammation. SolB transactivated human PXR , activated PXR in mice and upregulated hepatic expression of its downstream proteins, such as CYP3A11 , CYP2B10 and UGT1A1. SolB also significantly enhanced nuclear translocation of PXR and YAP in human cell lines. YAP signal pathway was activated by SolB in mice. Conclusion These findings demonstrated that SolB can significantly induce liver enlargement, which is associated with the activation of PXR and YAP pathways.
研究领域:
药理学
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分子生物学
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肝病学
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产品关联
DOI
PubMed
高影响因子
ACTA BIOCHIMICA ET BIOPHYSICA SINICA
·
2020
·Vol. 52
·期 8
·842-852
·
IF 2.836
Zhang Liang,Wang Xin,Liu Xin,Lv Mingyue,Shen Erdong,Zhu Guolian,Sun Zhe
Esophageal cancer (EC) is one of the most common aggressive malignant diseases worldwide. miR-28-5p plays important regulatory roles in many cancers including human EC. However, the molecular mechanism and potential role of miR-28-5p in EC remain uncertain. In this study, qRT-PCR and western blot analysis revealed that miR-28-5p expression was up-regulated and metastasis suppressor-1 (MTSS1) was down-regulated in EC tissues relative to matched para-cancer tissues. Cell counting kit-8 (CCK-8) assay demonstrated that miR-28-5p mimics increased cell viability, and miR-28-5p inhibitor decreased it. Flow cytometry (FCM) assay indicated that miR-28-5p mimics promoted cell cycle entry, while miR-28-5p inhibitor reduced it and induced cell apoptosis. Moreover, miR-28-5p mimics up-regulated the expressions of cyclin A, cyclin dependent kinase 2 (CDK2), cyclin D1, and cyclin E but down-regulated the expressions of cleaved caspase-3 and cleaved caspase-9, which was abolished by miR-28-5p inhibitor. Furthermore, luciferase reporter assay verified that miR-28-5p directly targeted MTSS1 3′UTR and down-regulated its expression. MTSS1 overexpression in TE-1 cells inhibited cell proliferation and promoted apoptosis induced by miR-28-5p mimics, whereas silencing of MTSS1 reversed cell progression induced by miR-28-5p inhibitor. We also demonstrated that miR-28-5p could promote esophageal tumor formation in vivo. Hematoxylin–eosin staining, immunohistochemistry, and TUNEL assays confirmed that miR-28-5p antagomir inhibited cell growth and accelerated apoptosis. Our results suggest that miR-28-5p may induce cell proliferation and suppress apoptosis to promote EC tumor formation via decreasing MTSS1 expression. Thus, miR-28-5p may be a potential target for human EC therapy.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2020
·
IF 4.872
Jiarong Qiu,Haiyan Yang,Yuting Shao,Longxiang Li,Shengwei Sun,Luyao Wang,Yuzhi Tan,Zhihong Xin
Our previous work has reported that EstJ6 was a phthalate-degrading hydrolase. In the study, a random mutant library was constructed by two rounds of error-prone PCR, three mutants (ET1.1, ET2.1, and ET2.2) with enhanced hydrolytic activity against dibutyl phthalate (DBP) were obtained. The best mutant ET2.2, accumulated three amino acid substitutions (Thr91Met, Ala67Val, and Val249Ile) and exhibited 2.8-fold increase enzyme activity and 2.3-fold higher expression level. Meanwhile, compared with EstJ6, ET2.2 showed over 50% improvement in thermostability (at 50 °C for 1 h) and 1.2-fold increase in 50% methanol tolerance. Kinetic parameters analysis revealed that the Km value for ET2.2 decreased by 60% and the kcat/Km value increased by 166%. The molecular docking indicated that the shortening of hydrogen bond between Ser146-OH and DBP-CO, which may led to an increase in enzyme activity and catalytic efficiency, the enhancement of hydrophobicity of hydrophobic pocket was related to the improvement of organic solvents tolerance, and three hydrophobic amino acid substitutions Thr91Met, Ala67Val, and Val249Ile facilitated to improve the thermal stability and organic solvents tolerance. These results confirmed that random mutagenesis was an effective tool for improving enzyme properties and lay a foundation for practical applications of phthalate-degrading hydrolase in biotechnology and industrial fields.
研究领域:
生物化学
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酶工程
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生物技术
·
环境科学
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DOI
PubMed
高影响因子
Biomed Research International
·
2019
·
IF 2.197
Dongyu Xie,Ping Zhou,Lin Liu,Wenjing Jiang,Haina Xie,Liang Zhang,Donghao Xie
Suitable content of iron is essential for human body, but iron overload is associated with many kinds of diseases including chronic liver damage. Recently, researchers find that iron overload promotes hepatocyte autophagy and apoptosis. However, the mechanism of iron overload in liver damage remains unclear. In this study, Lo2 cells were selected as the research object, iron dextran was a model drug, and astragaloside IV was a therapeutic drug to explore the role of iron overload. MTT assay and Annexin/PI double staining were used to measure cell viability and apoptosis. Ultrastructure was observed by transmission electron microscopy. The expression levels of apoptosis and autophagy-related proteins were determined by real-time PCR and Western Blot. The results showed that iron dextran could significantly inhibit Lo2 cell viability and increase the apoptosis rate, while astragaloside IV could reverse the inhibition of Lo2 cell viability and decrease the apoptosis rate. Transmission electron microscopy showed a significant increase in the number of autophagosomes after administration of iron dextran, and the application of astragaloside IV reduced the production of autophagosomes. LC3II/I was significantly upregulated in the model group but decreased in the astragaloside IV treatment group, and P62 showed the opposite trend. Iron dextran significantly upregulated the expression of Bax and downregulated Bcl2, while astragaloside IV reversed this trend. Finally, the inhibition of hepcidin caused by iron dextran was counteracted by astragaloside IV. In conclusion, the experimental results show that the iron overload model mainly induces excessive autophagy and apoptosis of hepatocytes, thus causing damage to hepatocytes, but astragaloside IV plays a certain therapeutic role in reversing this damage.
研究领域:
药理学
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肝病学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Journal of Materials Chemistry C
·
2018
·Vol. 6
·期 36
·9703-9712
·
IF 5.976
Qi You,Yang Chen
Gold nanoclusters (AuNCs) as a new class of luminescent materials have attracted a great deal of interest owing to their molecule-like properties and wide potential applications. However, limited successes have been achieved in producing intensely luminescent AuNCs. This paper reports a simple and convenient preparation method of strongly luminescent, highly heat-stable AuNCs by using functional ligands and a hydrothermal reaction. Adenosine cyclic monophosphate (cAMP) and tryptophan (Trp) are used as a template molecule and energy antenna ligand of AuNCs, respectively. By means of a common hydrothermal reaction, the resulting tryptophan-stabilized AuNCs (Trp–cAMP–AuNCs) generated surprising luminescence with a quantum yield of up to 59.6%; their luminescence is highly heat-stable, and they can be applied as a luminescent label in PCR (polymerase chain reaction) thermal cycles of DNA amplification. This unusually strong luminescence is attributed to the double effects of energy transfer from the ligand and hydrothermally-induced emission enhancement. The indole groups strongly interacting with gold atoms play an important role for the thermal-stability of the luminescence. The as-prepared AuNCs are also highly water-soluble and storage-stable. The present strategy offers a new way for and insight into the fabrication of strongly luminescent metal nanoclusters.
研究领域:
纳米技术
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材料科学
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化学
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生物技术
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产品关联
DOI
高影响因子
ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY
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2015
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IF 2.084
Xian-Hua Chen,Li Ma,Yi-Xiang Hu,Dan-Xian Wang,Li Fang,Xue-Lai Li,Jin-Chuan Zhao,Hai-Rong Yu,Hua-Zhong Ying,Chen-Huan Yu
Tris (2-ethylhexyl) trimellitate (TOTM) is commonly used as an alternative plasticizer for medical devices. But very little information was available on its biological effects. In this study, we investigated toxicity effects of TOTM on hepatic differential gene expression analyzed by using high-throughput sequencing analysis for over-represented functions and phenotypically anchored to complementary histopathologic, and biochemical data in the liver of mice. Among 1668 candidate genes, 694 genes were up-regulated and 974 genes were down-regulated after TOTM exposure. Using Gene Ontology analysis, TOTM affected three processes: the cell cycle, metabolic process and oxidative activity. Furthermore, 11 key genes involved in the above processes were validated by real time PCR. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that these genes were involved in the cell cycle pathway, lipid metabolism and oxidative process. It revealed the transcriptome gene expression response to TOTM exposure in mouse, and these data could contribute to provide a clearer understanding of the molecular mechanisms of TOTM-induced hepatotoxicity in human.
研究领域:
毒理学
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基因组学
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）