WATER RESEARCH
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2021
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IF 11.236
Hu Li,Ruiying Song,Yangyang Wang,Rongwei Zhong,Tiecheng Wang,Hanzhong Jia,Lingyan Zhu
Spread of antibiotic-resistant genes (ARGs) is a global public safety issue and inhibition their transfer is imperative. In this study, a novel strategy using environmental free radical exposure was developed to inhibit conjugative transfer of ARGs (RP4 plasmid) in aqueous solutions. Long-time free radical (·OH, 1 O 2 , and O 2 · − ) exposure significantly suppressed the conjugative transfer frequency of ARGs between Escherichia coli ( E. coli ) strains, and ·OH was more likely to attack ARG, thereby inhibiting the conjugate transfer frequency, compared to 1 O 2 and O 2 · − . Compared with the control, the conjugative transfer frequency significantly decreased from 4.08 × 10 −5 to 1.2 × 10 −8 after 10 min free radical exposure, confirming that the transfer and proliferation of ARGs were well inhibited. Correspondingly, the number of transconjugant significantly decreased by 61.7% after 10 min free radical exposure. Significant reductions in reactive oxygen species levels (ROS content and enzyme levels) and DNA damage-induced responses in the donor strains were observed after 10 min free radical exposure. Concurrently, intercellular contact was also weakened via inhibiting the synthesis of polysaccharides in extracellular polymeric substances. Moreover, the expressions of plasmid transfer genes were down-regulated after 10 min exposure due to the shortage of adenosine-triphosphate supply. This study firstly disclosed the underneath mechanisms for depressing ARGs transfer and dissemination via environmental free radical exposure.
研究领域:
微生物学
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环境科学
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分子生物学
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生物技术
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DOI
PubMed
高影响因子
Infection and Drug Resistance
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2022
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IF 4.177
Yingying Du,Shikui Mu,Yan Liu,Yinghua Yuan,Yunlou Zhu,Lijie Ma,Qixing Wang,Zhengfang Zhu,Yuhao Liu,Sheng Wang
Purpose This study retrospectively analyzed the genome characteristics of blaKPC-2 in multidrug-resistant Klebsiella pneumoniae collected from the ICU of a teaching hospital in Shanghai, China.Methods From February 2018 to December 2019, 36 strains of multidrug-resistant Klebsiella pneumoniae were collected from the bronchoalveolar lavage fluid of critically ill patients. The genome of all isolates was obtained through the Illumina sequence, and single nucleotide polymorphisms of the blaKPC-2 gene were analyzed to explore blaKPC-2’s evolutionary characteristics. Different strains’ genetic relationships and homology were studied by constructing an evolutionary tree on a single copy orthologue. Pacbio combined Illumina sequence was conducted to evaluate the structure and potential mobility of drug-resistant plasmids of the strain KP-s26.Results The distribution of resistance and virulence genes had little difference, but most strains had significant differences in the plasmid-encoded region. Most strains (31/36) carried the carbapenemase gene blaKPC-2, with no single nucleotide polymorphism in different strains. Extended-spectrum β-lactamase resistance genes, such as blaCTX-M and blaSHV, were found in the isolates, but no metallo-β-lactamases were detected. All strains with blaKPC-2 coexisted with chromosomal-associated fosfomycin resistance genes fosA6, and the coexistence of blaKPC-2 and blaCTX variants (blaCTX-M-15, blaCTX-M-65, and blaCTX-M-27) was also detected in 29/31 strains. The isolate KP-s26 carried five circular plasmids. pA and pB were conjugate plasmids, as they carried drug resistance genes and contained a complete IV secretion system.Conclusion The blaKPC-2 carbapenemase gene is relatively conservative in the process of evolution; drug-resistant plasmids containing conjugated transfer elements contribute to the spreading of drug resistance. The coexistence of blaKPC-2 with fosA6 or blaCTX-M variants was associated with increased fosfomycin resistance and broad-spectrum β-lactam resistance, respectively.Clinical Trials Registration Clinical Trials.gov Identifier: NCT03950544
研究领域:
微生物学
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基因组学
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传染病学
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DOI
PubMed
高影响因子
PHARMACOLOGICAL RESEARCH
·
2021
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IF 7.658
Yuqian Jia,Bingqing Yang,Jingru Shi,Dan Fang,Zhiqiang Wang,Yuan Liu
The widespread dissemination of antibiotic resistance genes (ARGs) is a serious problem and constitutes a threat for public health. Plasmid-mediated conjugative transfer of ARGs is recognized as one of the most important pathways accounting for this global crisis. Inhibiting the conjugative transfer of resistant gene-bearing plasmids provides a feasible strategy to prevent the spread of antibiotic resistance. Here we found that melatonin , a neurohormone secreted from pineal gland, substantially inhibited the horizontal transfer of RP4–7 plasmid in a dose-dependent manner. Furthermore, melatonin could also suppress the conjugal frequency of different types of clinical plasmids that carrying colistin resistance gene mcr-1 rather than bla NDM or tet (X) genes. Next, we investigated the mechanisms underlying the inhibitory effect of melatonin on conjugation. As a result, we showed that the addition of melatonin markedly reduced bacterial membrane permeability and inhibited the oxidative stress. In line with these observations, the conjugative transfer-related genes were regulated accordingly. Most importantly, we uncovered that melatonin disrupted bacterial proton motive force (PMF), which is an essential bacterial energy metabolism substance and is important for conjugative process. Collectively, these results provide implications that some non-antibiotics such as melatonin are effective inhibitors of transmission of ARGs and raise a promising strategy to confront the increasing resistant infections.
研究领域:
微生物学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
BIOCONJUGATE CHEMISTRY
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2024
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IF 4
Jun Liu,Yanhua Li,Kehui Zhou,Shijia Zhang,Yue Wang,Xiumei Wang,Xiabin Lan,Qixian Chen,Yan Zhao
Electrostatic self-assembly between negatively charged nucleic acids and cationic materials is the basis for the formulation of the delivery systems. Nevertheless, structural disintegration occurs because their colloidal stabilities are frequently insufficient in a hostile biological environment. To overcome the sequential biological barriers encountered during transcellular gene delivery, we attempted to use in situ polymerization onto plasmid DNA (pDNA) with a variety of functional monomers, including N-(3-aminopropyl)methacrylate, (aminopropyl)methacrylamide hydrochloride, 1-vinylimidazole, and 2-methacryloyloxyethylphosphorylcholine and N,N′-bis(acryloyl) cystamine. The covalently linked monomers could polymerize into a network structure on top of pDNA, providing excellent structural stability. Additionally, the significant proton buffering capacity of 1-vinylimidazole is expected to aid in the release of pDNA payloads from acidic and digestive endolysosomes. In addition, the redox-mediated cleavage of the disulfide bond in N,N′-bis(acryloyl)cystamine allows for the selective cleavage of the covalently linked network in the cytosolic microenvironment. This is due to the high intracellular level of glutathione, which promotes the liberation of pDNA payloads in the cell interiors. The proposed polymerization strategies resulted in well-defined nanoscale pDNA delivery systems. Excellent colloidal stabilities were observed, even when incubated in the presence of high concentrations of heparin (10 mg/mL). In contrast, the release of pDNA was confirmed upon incubation in the presence of glutathione, mimicking the intracellular microenvironment. Cell transfection experiments verified their efficient cellular uptake and gene expression activities in the hard-transfected MCF-7 cells. Hence, the polymerization strategy used in the fabrication of covalently linked nonviral gene delivery systems shows promise in creating high-performance gene delivery systems with diverse functions. This could open new avenues in cellular microenvironment engineering.
研究领域:
生物材料学
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基因治疗
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药物递送系统
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHYSICAL CHEMISTRY B
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2014
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IF 3.377
Long-Hai Wang,Sheng-Gang Ding,Jun-Jie Yan,Ye-Zi You
Encapsulation of negatively charged plasmid DNA into a small-sized nanocapsule without using any condensing agent is very challenging up to now. Here we report a versatile method for encapsulating large-sized plasmid DNAs into small-sized bioreducible nanocapsules in which shearing force and surfactant can fold large-sized plasmid DNAs into small-sized emulsion droplets containing bioreducible branched polymers. Subsequently, temperature triggers the bioreducible branched polymers to aggregate and cross-link at the water/oil interface of the emulsion nanodroplet, forming a bioreducible shell around the nanodroplet. Thus, a small-sized nanocapsule (∼110 nm) containing large-sized plasmid DNA (∼1900 nm long) forms by removal of the surfactant.
研究领域:
生物材料学
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基因治疗
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药剂学
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DOI
高影响因子
Journal of Nanomaterials
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2017
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IF 1.871
Juan Zhang,Huanhuan Weng,Xiangwan Miao,Quanming Li,Siqi Wang,Huifen Xie,Tao Liu,Minqiang Xie
A novel folate (FA) receptor-targeted superparamagnetic Fe3O4 nanoparticles (SPIONs) codelivering cisplatin (CDDP) and tissue factor pathway inhibitor-2 (TFPI-2) plasmid DNA (pDNA) was constructed. The core shell nanocomposites ([email protected]) were composed of superparamagnetic Fe3O4 core that binds CDDP and TFPI-2 shell that combines with folate-polyethylene glycol-polyethyleneimine (FA-PEG-PEI) via electrostatic interaction. The shell containing FA-PEG-PEI and TFPI-2 plasmid was synthesized through amidation reaction and electrostatic adsorption and the core containing SPION-CDDP was modified by aldehyde sodium alginate. Proton nuclear magnetic resonance and Fourier transform infrared spectra on FA-PEG-PEI polymers showed characteristic peaks of various metabolites in corresponding frequency. Transmission electron microscopy image of [email protected] nanoparticles demonstrated a near-monodisperse spherical morphology, while dynamic light scattering studies indicated an intensity-average diameter of 149.5 nm. Zeta potential was 14.89 ± 1.83 mv and the final concentration of loaded CDDP was 100 ug/ml. Gel electrophoresis data showed that the nanocomposite would protect TFPI-2 pDNA from being digested by DNases. Compared with CNE-2 cells, the good targetability and better gene transfection efficiency (57.9%) were detected by Prussian blue iron stain and fluorescence analysis in HNE-1 cells in vitro. The results suggested the potential application of [email protected] as a multifunctional anticancer nanomedicine on targeting therapy for FR positive NPC.
研究领域:
纳米技术
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癌症治疗
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药学
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生物医学工程
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产品关联
DOI
高影响因子
WATER RESEARCH
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2025
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IF 12.4
Lingling Rong,Baoyin Zhang,Hongmu Qiu,Hailin Zhang,Jiayin Yu,Qing Yuan,Ligui Wu,Hong Chen,Yuanmin Mo,Xiaoming Zou,Xubiao Luo
The generational effects of antibiotics are well-documented within the field of pharmacology, but their specific influence on the enhancement of plasmid-mediated conjugative transfer remains insufficiently understood, although plasmid-mediated conjugative transfer represents a crucial process for the horizontal dissemination of antibiotic resistance genes (ARGs). We pioneeringly investigate the effects of eight tetracyclines (TCs) across three generations on the conjugative transfer of RP4 plasmid, utilizing Escherichia coli strains isolated from typical wastewater. Results indicate that TCs enhance plasmid conjugative transfer, exhibiting significant generational effects. Average hormetic zone for conjugative transfer (ΔA R ) induced by 1st-generation TCs is 4.46 to 5.95 times greater than that induced by 2nd- and 3rd-generation TCs, respectively. Furthermore, this intriguing phenomenon is unrelated to TCs’ degradation characteristics (P > 0.05), but is closely associated with bacterial biochemical responses (P < 0.05), including reactive oxygen species (ROS), cell membrane permeability, and ATP synthesis. Moreover, mechanistic results reveal that key structural parameters of TCs significantly impact their interactions with the essential protein UspD, which regulates biochemical processes and relevant genes on RP4 plasmid in donor bacteria, thereby substantially enhancing the plasmid conjugative transfer frequency and exhibiting significant generational effects (P < 0.001). Our findings shed light on the generational effects of TCs on plasmid conjugative transfer in wastewater treatment systems, offering theoretical supports for ARGs dissemination control.
研究领域:
环境科学
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微生物学
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药理学
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分子生物学
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水处理
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抗生素抗性
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2025
·Vol. 23
·期 1
·1-25
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IF 12.6
Zhang Xiaoyu,Guo Qianping,Fang Jiawei,Cheng Qi,Zhu Zhuang,Yu Qifan,Wang Huan,Hong Youzhi,Liu Chengyuan,Yang Huilin,Zhu Caihong,Li Bin,Ni Li
Nucleus pulposus cells (NPCs) undergo metabolic disorders and matrix pathological remodeling under the influence of various adverse factors during intervertebral disc degeneration (IVDD), whereas post-translational modifications (PTMs) can confer cells with the capacity to respond quickly and adapt to complex environmental changes. Here, SIRT1 protein, a key regulator within PTMs framework, was applied against the hostile degenerative microenvironment. Then, it was sequentially assembled with SOX9-expressing plasmid, an essential transcription factor to promote extracellular matrix (ECM) biosynthesis, onto a phenylboronic acid-functionalized G5-dendrimer to construct a multifunctional nanoplatform for IVDD therapy. In vitro, the nanoplatforms showed antioxidant capacity, and the ability to restore mitochondrial homeostasis and normal ECM metabolism, as well as to maintain cellular phenotypes. RNA sequencing suggested that inhibition of the Nod-like receptor signaling might be the mechanism behind their therapeutic effects. The nanoplatforms were then wrapped in a designed dynamic hydrogel, not only prolonging the retention time of the loaded cargoes, but also well maintaining the disc structure, height, and water content in vivo. Overall, this study presents a convenient assembled strategy to inhibit the multiple adverse factors, and hold promise for the IVDD treatment. Graphical abstract
研究领域:
医学
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生物材料学
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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
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2023
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IF 13.6
Yan-Zi Wang,Xin-Li An,Xiao-Ting Fan,Qiang Pu,Hu Li,Wen-Zhen Liu,Zhuo Chen,Jian-Qiang Su
Inhibition of plasmid transfer, including transformation and conjugation, is essential to prevent the spread of plasmid-encoded antimicrobial resistance. Photosensitizers have been successfully used in the treatment of serious infectious diseases, however, the effects of photosensitizers on the plasmid transfer are still elusive. In this study, we determined the transformation and conjugation efficiency of plasmid pUC19 and pRP4, respectively, when exposed to a photosensitizer (Visible Light-activated Rose Bengal, VLRB). The results showed that the activation of VLRB resulted in up to a 580-fold decrease in the transformation frequency of pUC19 and a 10-fold decrease in the conjugation frequency of pRP4 compared with the non-VLRB control. The inhibition of pUC19 transformation by VLRB exhibited a dose-dependent manner and was attributed to the changes in the plasmid conformation. The inhibition of pRP4 conjugation was associated with the generation of extracellular free radicals , induced oxidative stress , suppression of the mating pair formation gene ( trbBp ) and DNA transfer and replication gene ( trfAp ), and enhanced expression of the global regulatory genes ( korA , korB , and trbA ). These findings highlight the potential of visible light-activated photosensitizer for mitigating the dissemination of plasmid-encoded antibiotic resistance genes.
研究领域:
微生物学
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分子生物学
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感染病学
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产品关联
DOI
PubMed
高影响因子
Transboundary and Emerging Diseases
·
2024
·Vol. 2024
·期 1
·7063673
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IF 3.5
Yuqian Jia,Zhiwan Zheng,Bingqing Yang,Haijie Zhang,Zhiqiang Wang,Yuan Liu
The dissemination of antimicrobial resistance (AMR) severely degrades the performance of antibiotics and constantly paralyzes the global health system. In particular, plasmid-mediated transfer of antibiotic resistance genes (ARGs) across bacteria is recognized as the primary driver. Therefore, antiplasmid transfer approaches are urgently warranted to resolve this intractable problem. Herein, we demonstrated the potential of azidothymidine (AZT), an FDA-approved anti-HIV drug, as a broad-spectrum horizontal transfer inhibitor to effectively prevent the transmission of multiple ARGs, including mcr-1 , bla NDM−5 , and tet (X4), both in vitro and in vivo . It was also noteworthy that the inhibitory effect of AZT was proved to be valid within and across bacterial genera under different mating conditions. Mechanistic studies revealed that AZT dissipated bacterial proton motive force, which was indispensable for ATP synthesis and flagellar motility. In addition, AZT downregulated bacterial secretion systems involving general and type IV secretion systems (T4SS). Furthermore, the thymidine kinase, which is associated with DNA synthesis, turned out to be the potential target of AZT. Collectively, our work demonstrates the broad inhibitory effect of AZT in preventing ARGs transmission, opening new horizons for controlling AMR.
研究领域:
微生物学
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药理学
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传染病学
✓ 已验证
DOI
高影响因子
Frontiers in Microbiology
·
2025
·
IF 4.5
Zhuocheng Yao,Yuhan Yang,Yanchun Gong,Shiyi Shi,Yunying Ge,Weiliang Zeng,Deyi Zhao,Jianming Cao,Tieli Zhou,Mo Shen
Disinfectants are commonly utilized by humans to combat microorganisms. However, residual disinfectants may promote environmental antimicrobial resistance by facilitating horizontal gene transfer (HGT) of antibiotic resistance genes. Bronopol is a routinely used disinfectant that persists in the environment, and previous studies have concentrated on its ecotoxicity rather than its implications on the propagation of resistance genes. This study aimed to establish an in vitro conjugation model to investigate whether bronopol promotes the transfer of antibiotic resistance genes (ARGs) via plasmid conjugation. Using Escherichia coli DH5α and DC8855 as donors harboring RP4-7 and blaNDM-4-positive IncFII(K) plasmids, respectively, and J53 as the recipient strain, we found that sub-inhibitory concentrations of bronopol (2 μg/L and 20 μg/L) significantly increased the conjugative transfer frequency (CTF) of both plasmids. Mechanistic analysis revealed that bronopol enhanced bacterial membrane permeability, as demonstrated by propidium iodide (PI) staining, 1-N-phenylnaphthylamine (NPN) fluorescent probes, transmission electron microscopy (TEM), and upregulation of the outer membrane protein gene ompC. Additionally, bronopol treatment upregulated RP4 plasmid-encoded genes involved in DNA transfer/replication (trfAp) and the global regulator of HGT (kilA/kilB). These findings highlight a previously unrecognized role of bronopol in facilitating the dissemination of antibiotic resistance genes, particularly those of clinical significance.
研究领域:
微生物学
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环境科学
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遗传学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
·
2015
·
IF 3.082
Dongyang Xu,Qing Nie,Wenyan Wang,Lei Shi,He Yan
In this study, we characterized a bcrABC cassette and its genetic environment harbored by a plasmid in Listeria monocytogenes ( L . monocytogenes ) 11GZL18, a strain isolated from raw meat in 2011. The bcrABC cassette and its genetic environment were characterized, with a total of 33,727 nt nucleotide sequence obtained. The nucleotide sequences of the bcrABC cassette in strain 11GZL18 exhibited 100% identity to that on plasmid pLM80, which is harbored by L . monocytogenes strain H7550and H7858, and the neighboring 21,678 nt nucleotide sequence of bcrABC cassette showed 99% identity with plasmid pLM80. The plasmid curing experiment demonstrated the role of the plasmid in conferring benzalkonium chloride (BC) and cadmium (Cd) tolerance in this strain. The bcrABC cassette and cadAC genes from the L . monocytogenes 11GZL18 were harbored by plasmid, functional and transmissible, and led to the acquired tolerance in Gram-negative Escherichia coli ( E . coli ) DH5α by chemical and natural transformation. Besides, the efflux pump activity that is conferring tolerance to BC and Cd was observed in strain 11GZL18, while not in a plasmid-cured strain 11GZL18-C, confirming that efflux pumps play a role in plasmid-mediated tolerance to BC and Cd in L . monocytogenes 11GZL18. In this study we characterized the genetic organization of a novel BC and Cd tolerance determinants-harboring plasmid in a L . monocytogenes strain isolated from raw meat of animal origin, and demonstrated the potential horizontal transferability of this bcrABC cassette-harboring plasmid to E . coli . The findings will further improve our understanding of the adaptations of this organism to disinfectants such as BC and may contribute to elucidating possible dissemination of BC tolerance in foodborne L . monocytogenes .
研究领域:
微生物学
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食品安全
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分子生物学
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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）