JOURNAL OF APPLIED MICROBIOLOGY
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2022
·Vol. 133
·期 5
·3161-3175
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IF 4.059
Pan Deng,Wang Xing‐Yun,Zhou Jin‐Wei,Yang Liang,Khan Abbas,Wei Dong‐Qing,Li Jun‐Jian,Jia Ai‐Qun
AimsIn the current study the anti‐virulence and anti‐biofilm activities of the cinnamic acid derivative, 3‐methoxycinnamic acid, was investigated against Agrobacterium tumefaciens.Methods and ResultsBased on the disc diffusion test and β‐galactosidase activity assay, 3‐methoxycinnamic acid was shown to interfere with the quorum sensing (QS) system of A. tumefaciens. Crystal violet staining assay, phenol‐sulfuric acid method, Bradford protein assay and confocal laser scanning microscopy (CLSM) revealed that the biofilm formation of A. tumefaciens was inhibited after the treatment of 3‐methoxycinnamic acid. Employing high‐performance liquid chromatography (HPLC) analysis of culture supernatant revealed that the production of 3‐oxo‐octanoylhomoserine lactone (3‐oxo‐C8‐HSL) decreased concentration‐dependently after treatment with 3‐methoxycinnamic acid. Swimming and chemotaxis assays also indicated that 3‐methoxycinnamic acid had a good effect on reducing the motility and chemotaxis of A. tumefaciens. In addition, the RT‐qPCR, molecular docking and simulations further demonstrated that 3‐methoxycinnamic acid could competitively inhibit the binding of 3‐oxo‐C8‐HSL to TraR and down‐regulate virulence‐related genes.Conclusions3‐Methoxycinnamic acid is proved to have good anti‐virulence and anti‐biofilm activities against A. tumefaciens.Significance and Impact of the StudyThis is the first study that investigates the anti‐virulence and anti‐biofilm activities of 3‐methoxycinnamic acid against A. tumefaciens. With its potential QS‐related virulence and biofilm inhibitory activities, 3‐methoxycinnamic acid is expected to be developed as a potent pesticide or adjuvant for the prevention and treatment of crown gall caused by A. tumefaciens.
研究领域:
微生物学
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植物病理学
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药理学
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DOI
PubMed
高影响因子
Analytical Methods
·
2023
·Vol. 15
·期 26
·3240-3250
·
IF 3.1
Ruihua Tang,Xueyan Yan,Min Li,Aoqi Du,Hui Yang,Huancai Yin,Mingyue Xie
Nucleic acid detection technologies have been widely utilized for various diseases. Conventional laboratory tests are less suitable for use in resource-limited settings as they are time-consuming, high-cost, complex, and heavily dependent on benchtop equipment. Rapid nucleic acid detection methods that consist of rapid nucleic acid extraction steps could overcome these challenges. A paper-based platform has been utilized to develop various rapid nucleic acid extraction methods owing to its cost-effectiveness, portability, and easy-modification. However, the existing paper-based nucleic acid extraction technologies mainly focus on improving the adsorption capacity of nucleic acids without reducing the non-specific adsorption capacity of proteins. In this study, paper-based nucleic acid extraction technology with wash-free, elution-free, and low protein adsorption was developed. The fabrication of paper involves the mixing of polyethylene glycol (PEG)-modified cotton fiber, chitosan (COS)-modified cotton fiber, and cotton fiber to form PEG-modified cotton fiber/chitosan-modified cotton fiber/cotton fiber (PEG-CF/COS-CF/CF) paper by the wet molding method. The result showed that PEG-CF/COS-CF/CF paper has a desirable pore size (23.9 ± 4.03 μm), good mechanical strength (dry: 9.37 Mpa and wet: 0.28 Mpa), and hydrophilicity (contact angle: 42.6° ± 0.36°). NH3+ groups of COS and OH− groups of PEG were observed on its surface and the adsorption efficiency of nucleic acid in TE buffer was 42.48% ± 0.30%. The limit of detection of pure DNA with this PEG-CF/COS-CF/CF paper by qPCR was as low as 25 ng. Additionally, this platform could successfully extract nucleic acid from 30 μL of a saliva sample, highlighting its potential use for clinical sample testing. The proposed paper-based nucleic acid extraction platform shows tremendous potential for disease diagnosis in resource-limited settings.
研究领域:
生物技术
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分子生物学
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诊断技术
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材料科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2023
·
IF 13.6
Shunlan Li,Dongdong Chen,Ziqing Liu,Shipin Tao,Tianyi Zhang,Yiwen Chen,Lingzhi Bao,Jie Ma,Yuee Huang,Shengmin Xu,Lijun Wu,Shaopeng Chen
Antibiotic abuse is the main reason for the drug resistance of pathogenic bacteria, posing a potential health risk. Antibiotic surveillance is critical for preventing antibiotic contamination. This study aimed to develop a sensitive and broad-spectrum whole-cell biosensor for tetracycline antibiotics (TCs) detection. Wild-type TCs-responsive biosensor was constructed by introducing a tetracycline operon into a sfGFP reporter plasmid. Using error-prone PCR, mutation libraries containing approximately 10 7 variants of the tetracycline repressor (TetR) gene were generated. The tigecycline-senstive mutants were isolated using high-throughput flow cytometric sorting. After 2 rounds of directed evolution, a mutant epS2–22 of TerR was isolated and assembled as a TCs biosensor. The epS2–22 biosensor was more sensitive and broad-spectrum than the wild-type biosensors. The detection limits of the epS2–22 biosensor for seven TCs were 4- to 62-fold lower than the wild-type biosensor (no response to tigecycline). Meanwhile, the epS2–22 biosensor had a shorter detection time and a stronger signal output than the wild type. In addition, the evolved epS2–22 biosensor showed excellent performance in detecting low traces of TCs in environmental water. These results suggest that directed evolution is a powerful tool for developing high-performance whole-cell biosensors, and the evolved epS2–22 biosensors have the potential for wider applications in real-world TCs detection.
研究领域:
生物技术
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微生物学
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环境科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 22
·11967
·
IF 4.9
Yuzhen Tian,Xizi Wang,Huoqing Huang,Xin Deng,Baihong Zhang,Yixuan Meng,Libo Wu,Hang Chen,Yun Zhong,Wenli Chen
Citrus Huanglongbing (HLB) is the most destructive citrus disease worldwide. The etiological agent responsible for this disease is “CandidatusLiberibacter asiaticus” (CLas), a phloem-restricted bacterium transmitted by psyllid vectors. To date, effective practical strategies for curing Citrus HLB remain elusive. Additionally, no susceptibility genes associated with HLB have been identified in Citrus species, thereby complicating the application of gene-editing techniques such as CRISPR-Cas9 to enhance resistance to HLB. The co-chaperoneDnaJplays a crucial role in protein folding and the regulation of various physiological activities, and it is also associated with multiple pathological processes.DnaJhas been extensively studied in many species, including Arabidopsis, rice, and wheat. However, there is limited information available regarding the DnaJ gene family in citrus. In this study, we conducted a comprehensive genome-wide analysis of the DnaJ family genes in various Citrus species. The Citrus genome was identified to contain 86DnaJgenes, which were unevenly distributed across nine chromosomes. Phylogenetic analysis indicated that these genes could be classified into six distinct groups. Furthermore, transcriptomic analysis revealed that nineDnaJgenes exhibited significantly higher induction in HLB-infected samples relative to non-HLB-infected Citrus.Cis-acting elements within the promoters ofDnaJgenes were also examined, revealing the presence of hormone and defense/stress responsiveness elements (TC-rich) distributed on theClDJC24gene. The results were validated using quantitative real-time PCR (qRT-PCR). Additionally, the silencing ofClDJC24suggested that this gene negatively regulates disease resistance in Citrus. Our study provided useful clues for further functional characterization and constructed a theoretical foundation for disease-resistant breeding in Citrus.
研究领域:
植物病理学
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分子生物学
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基因组学
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植物遗传学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2025
·
IF 7.7
Xiaolong Guo,Yan Zhao,Xuemei Xu,Zhibin Wu,Li Xiao,Xiaoyang Ma,Ziqiu Lei,Yilin Zhao,Chengli Zhang,Lixing Song,Minggang Yin
Rapid and accurate detection of Porphyromonas gingivalis ( P. gingivalis ) is crucial for the timely diagnosis and management of periodontal diseases as well as for elucidating its role in systemic conditions. However, conventional diagnostic methods are complex, time-consuming, and unsuitable for point-of-care testing, necessitating the development of a more efficient and accessible detection system. This study presents a novel dual-probe fluorescence platform featuring cadmium telluride quantum dots (CdTe QDs) and a specially designed probe (PAN) as fluorescent molecules, which does not require a washing step. The PAN and CdTe QD probes can efficiently differentiate between Cu 2 + and Cu + ions through ionic hydrolysis and cation exchange reactions, respectively, resulting in the sensitive and rapid detection of arginine-specific gingipain, a key biomarker for P. gingivalis . Furthermore, the color change of CdTe QDs upon contact with saliva samples facilitates the visual identification of P. gingivalis . This dual-probe platform demonstrated a higher signal-to-noise ratio and greater reliability compared to the single-probe methods. Moreover, this platform features a low detection limit of 5 and 20 colony-forming units (CFUs)/mL for the PAN and CdTe QDs signals, respectively, with a visual detection threshold as low as 200 CFU/mL. Thus, this platform provides quantitative and qualitative results that are 100 % in concordance with quantitative PCR results from clinical saliva samples of patients with periodontitis and healthy controls. This simple, rapid, and cost-effective diagnostic tool can be potentially used for point-of-care applications, enabling early detection and improved monitoring of oral health and related diseases.
研究领域:
生物医学工程
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微生物学
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牙科医学
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生物传感器
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纳米技术
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诊断检测
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产品关联
DOI
高影响因子
ACS Sensors
·
2025
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IF 9.1
Lijuan Huang,Bing Du,Xiaohu Cui,Meiqi Liu,Zhoufei Li,Xiaoyun Cui,Ziying Xu,Zihui Yu,Shuo Zhao,Yanling Feng,Hanqing Zhao,Jianhai Long,Jing Yuan
Klebsiella pneumoniae (Kpn) is a common opportunistic pathogen, with hypervirulent strains (hvKpn) posing significant risks even to healthy individuals. Traditional clinical diagnostics often use real-time quantitative polymerase chain reaction (PCR) with TaqMan probes, which are costly and unstable due to their reliance on fluorophore and quencher molecules. This study introduces a novel probe using silicon-doped carbon quantum dots (Si-QDs) and single-layer graphene oxides (GOs), forming the Si-QD-probe-GO. This new probe offers excellent selectivity, sensitivity, stability, low photobleaching, and cost-effectiveness. A sensor array targeting khe, peg-344, and iucA genes is developed to detect Kpn and differentiate between classical Kpn (cKpn) and hvKpn. Using a partial least-squares discriminant analysis (PLS-DA) model, the system achieves 100% accuracy in distinguishing hvKpn from cKpn. The study also employs droplet digital PCR (ddPCR) with microfluidic technology to enhance sensitivity and quantification, achieving a detection limit of 5 CFU/mL. In a mouse pneumonia model, the PLS-DA model accurately identifies hvKpn in lung samples. The ddPCR quantification using Si-QD-probe-GO shows strong correlation with traditional culture methods. This research presents a promising ddPCR system for precise detection and virulence identification of Kpn, offering a new direction for clinical diagnostics.
研究领域:
微生物学
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分子诊断
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纳米技术
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生物医学工程
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传染病学
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产品关联
DOI
PubMed
高影响因子
FOOD AND CHEMICAL TOXICOLOGY
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2021
·
IF 6.025
Yue Leng,Li Ren,Shu Niu,Tiehua Zhang,Jie Zhang
The endocrine disruptive capability of plasticizers to activate nuclear receptors has attracted great interest. This study is aimed to assess the potential glucocorticoid effects of metabolites of plasticizers. The effects of metabolites of plasticizers on the transcriptional activity of glucocorticoid receptor (GR) were investigated using reporter gene assays. All of them failed to exhibit agonistic/antagonistic effects on GR. However, a combination of dexamethasone and monobutyl phthalate (MBP) could synergistically activate GR. MBP combined with dexamethasone also enhanced GR nuclear translocation by Western blot , while mifepristone restrained GR cytoplasmic-to-nuclear translocation. MBP co-treated with dexamethasone resulted in synergistic induction of PEPCK and MKP-1 gene expression by real-time PCR and PEPCK protein level by Western blot. Furthermore, the carboxyl and ester groups of MBP have influences on the charge distribution of MBP, leading to change of electrostatic interactions between MBP and GR by calculations on electronic properties. Both hydrophobic and hydrogen bonding interactions play a crucial role in the stabilization between MBP and GR conducted by molecular docking and dynamics simulation. This work confirms that GR could remain stable upon binding to MBP. In conclusion, dexamethasone and MBP could synergistically activate GR, resulting in synergetic enhancement of subsequent GR-mediated endocrine disrupting effect.
研究领域:
毒理学
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内分泌学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
Theranostics
·
2017
·
IF 8.766
Feifei Xu,Weixian Zhou,Jianxiang Cao,Qingqing Xu,Dechen Jiang,Yun Chen
The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarker. There is much evidence indicating that a panel of miRNAs, termed “miRNA fingerprints”, is more specific and informative than an individual miRNA as biomarker. Thus, multiplex assays for simultaneous quantification of multiple miRNAs could be more potent in clinical practice. However, current available assays normally require pre-enrichment, amplification and labeling steps, and most of them are semi-quantitative or lack of multiplexing capability. In this study, we developed a quasi-targeted proteomics assay for multiplexed miRNA quantification by a combination of DNA-peptide probes and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Specifically, the signal of target miRNAs (i.e., miR-21, miR-let7a, miR-200c, miR-125a and miR-15b) was converted into the mass response of reporter peptides by hybridization of miRNAs with DNA-peptide probes and subsequent tryptic digestion to release the peptides. After a careful optimization of conditions related to binding, conjugation, hybridization and multiple reaction monitoring (MRM) detection, the assay was validated for each miRNA and the limit of quantification (LOQ) for all the miRNAs can achieve 1 pM. Moreover, crosstalk between DNA-peptide probes in multiplex assay was sophisticatedly evaluated. Using this quasi-targeted proteomics assay, the level of target miRNAs was determined in 3 human breast cell lines and 36 matched pairs of breast tissue samples. Finally, simplex assay and qRT-PCR were also performed for a comparison. This approach grafts the strategy of targeted proteomics into miRNA quantification and may offer a new way for multiplexed miRNA profiling.
研究领域:
癌症生物学
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蛋白质组学
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分子生物学
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生物化学
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临床诊断
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产品关联
DOI
PubMed
高影响因子
AQUATIC TOXICOLOGY
·
2016
·
IF 3.557
Jieyi Li,Jinfeng Liu,Yuhuan Zhang,Xuedong Wang,Weijun Li,Hongqin Zhang,Huili Wang
The toxicity of β-diketone antibiotics (DKAs) to larval and adult zebrafish ( Danio rerio ) was investigated by miRNA sequencing and bioinformatics analyses. In control and DKA-exposed groups, 215 differentially expressed miRNAs were screened, and 4076 differential target genes were predicted. Among 51 co-differentially expressed genes, 45 were annotated in KOG functional classification, and 34 in KEGG pathway analysis. The homology analysis of 20 miRNAs with human hsa-miRNAs demonstrated 17 high homologous sequences. The expression levels of 12 miRNAs by qRT-PCR were consistent with those by sRNA-seq. A regulatory network for 4 positive miRNA genes (dre-miR-10, ⿿96, ⿿92 and ⿿184) was plotted, and the high-degree of connectivity between miRNA-gene pairs suggests that these miRNAs play critical roles during zebrafish development. The consistent expression of dre-miR-184 and dre-miR-96 was proved in 120-hpf zebrafish brain, gill, otoliths and lateral line neuromast by qRT-PCR, miRNA-seq, W-ISH and ISH. DKA-exposure led to vacuolation of interstitial cells, reduced number of neurons, glial cell proliferation and formation of glial scar, and the obvious abnormality of cell structure might result from abnormal expression of differentially expressed miRNAs. In general, chronic DKA-exposure resulted in comprehensively toxic effects on larval and adult zebrafish tissues, especially for nervous system.
研究领域:
水生毒理学
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分子生物学
·
生物信息学
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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF ORAL BIOLOGY
·
2020
·
IF 1.931
Ying Bian,Juan Xiang
Background Osteogenic differentiation of human periodontal ligament cells (hPDLCs) is crucial for regenerate periodontal tissues. In this study, we investigated the function of salvianolic acid B (Sal B) in osteogenesis of hPDLCs. Methods HPDLCs were isolated from healthy third molar roots. HPDLCs at passage 3 were identified by morphological observation and immunohistochemistry of vimentin . The viability of hPDLCs incubated with Sal B at concentrations of 0μM, 0.1μM, 0.5μM, 1μM and 5μM were measured by CCK-8 assay. To evaluate the effect of Sal B on osteogenic differentiation of hPDLCs, the alkaline phosphatase (ALP) activity, osteogenic differentiation markers, and mineralized nodules were determined by ALP kit, qRT-PCR and alizarin red S staining, respectively. To confirm the function of Sal B in hPDLCs involved in Wnt/β-catenin signaling pathway , hPDLCs were incubated with Sal B or co-incubated with Sal B and DKK-1 (a inhibitor of Wnt/β-catenin). The levels of Wnt/β-catenin signaling pathway and osteogenic differentiation-associated indicators were then determined. Results HPDLCs showed a typical fibroblast-like and spindle-shaped, with vimentin-positive. The viability of hPDLCs had no obvious change with stimulation of Sal B at various doses. Sal B promoted the increase of ALP activity, osteogenic differentiation markers levels, mineralized nodules and activation of Wnt/β-catenin signaling pathway, and DKK-1 could block those effects of Sal B on hPDLCs. Conclusion Sal B promoted osteogenesis of hPDLCs through Wnt/β-catenin signaling pathway, which providing a potential drug for periodontitis treatment .
研究领域:
牙科学
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药理学
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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 HAZARDOUS MATERIALS
·
2022
·
IF 14.224
Xinyi Hu,Yulong Fu,Hongyu Shi,Weijian Xu,Chaofeng Shen,Baolan Hu,Liping Ma,Liping Lou
Various antibiotic resistant bacteria (ARB) can thrive in soil and resist such environmental pressures as antibiotics through cooperative resistance, thereby promoting ARB retention and antibiotic resistance genes transmission. However, there has been finite knowledge in regard to the mechanisms and potential ecological risks of cooperative resistance in soil microbiome. In this study, soil minerals and organic matters were designed to treat a mixture of two Escherichia coli strains with different antibiotic resistance ( E. coli DH5α/pUC19 and E. coli XL2-Blue) to determine how soil components affected cooperative resistance, and Luria-Bertani plates containing two antibiotics were used to observe dual-drug resistant bacteria (DRB) developed via cooperative resistance. Results showed quartz, humic acid , and biochar promoted E. coli XL2-Blue with high fitness costs, whereas kaolin, montmorillonite , and soot inhibited both strains. Using fluorescence microscope and PCR, it was speculated DRB could resist the antibiotic pressure via E. coli XL2-Blue coating E. coli DH5α/pUC19. E. coli DH5α/pUC19 dominated cooperative resistance. Correlation analysis and scanning electron microscope images indicated soil components influenced cooperative resistance. Biochar promoted cooperative resistance by increasing intracellular reactive oxygen species , thereby reducing the dominant strain concentration required for DRB development. Kaolin inhibited cooperative resistance the most, followed by soot and montmorillonite.
研究领域:
微生物学
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环境科学
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土壤学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2020
·
IF 6.551
Chenhui Duan,Yanjun Fang,Jingran Sun,Zhenxin Li,Qiangqiang Wang,Jialei Bai,Hui Peng,Jun Liang,Zhixian Gao
The health risks of exposure to plasticizers have received widespread attention, however, little is known about the effects of fast food packaging plasticizers on steroid hormone synthesis. In the present study, the types and migration of plasticizers in some commonly used fast-food packaging materials were detected by GC–MS, and the interference effects of these plasticizers and their metabolites on steroid hormone synthesis in the human body were evaluated by the H295R steroidogenesis assay. The GC–MS results showed that the main plasticizer compounds that migrated from fast food packaging into food were di(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) and bis(2-ethylhexyl) adipate (DEHA). Exposure to these chemicals (100–1000 μM) can significantly reduce the viability of H295R cells in a dose-response manner, and these plasticizers and their metabolites that migrated into oily foods at high temperatures (0.25–25 μM) could significantly increase the E2 level and reduce the T level in H295R cells. According to the qRT-PCR data, 0.25 to 25 μM mono(2-ethylhexyl) phthalate (MEHP) significantly upregulated the expression levels of 17β-HSD1 and CYP19A1, and downregulated those of CYP17A1, CYP11A1 and StAR. The Western blot results were consistent with those of qRT-PCR. In summary, these results indicated that even exposure to low concentrations (≤1 mg/l or 2.5 μM) of these chemicals and their metabolites can cause significant endocrine-disrupting effects.
研究领域:
环境健康
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毒理学
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内分泌学
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）