INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2024
·Vol. 25
·期 15
·8479
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IF 4.9
Yuning Peng,Huanrong Tang,Hao Xiao,Wenyuan Chen,Jingdong Song,Jing Zheng,Hongrong Liu
T5 is a siphophage that has been extensively studied by structural and biochemical methods. However, the complete in situ structures of T5 before and after DNA ejection remain unknown. In this study, we used cryo-electron microscopy (cryo-EM) to determine the structures of mature T5 (a laboratory-adapted, fiberless T5 mutant) and urea-treated empty T5 (lacking the tip complex) at near-atomic resolutions. Atomic models of the head, connector complex, tail tube, and tail tip were built for mature T5, and atomic models of the connector complex, comprising the portal protein pb7, adaptor protein p144, and tail terminator protein p142, were built for urea-treated empty T5. Our findings revealed that the aforementioned proteins did not undergo global conformational changes before and after DNA ejection, indicating that these structural features were conserved among most myophages and siphophages. The present study elucidates the underlying mechanisms of siphophage infection and DNA ejection.
研究领域:
病毒学
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结构生物学
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微生物学
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高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Huilei Dong,Hongfei Chen,Juqian Jiang,Hui Zhang,Chenxin Cai,Qingming Shen
Sensitive and specific detection of tumor exosomes is of great significance for early cancer diagnosis. In this paper, we report an aptamer strategy for exosome detection based on aptamer recognition-induced multi-DNA release and cyclic enzymatic amplification. First, we use aptamer–magnetic bead bioconjugates to capture tumor exosomes derived from LNCaP cells, leading to the release of three kinds of messenger DNAs (mDNAs). After magnetic separation, the released mDNAs hybridized with the probe DNAs immobilized on a gold electrode. Electroactive Ru(NH3)63+ was used as the signal reporter because of its electrostatic attraction to DNA. Subsequent Exo III cyclic digestion caused the electrochemical signal to “turn off”. Because the electrochemical signal reflects the concentration of Ru(NH3)63+ and the concentration of Ru(NH3)63+ is correlated with the mDNA concentration, which is correlated with the exosome concentration, the tumor exosomes can be detected by examining the decrease in the peak current of Ru(NH3)63+. In this paper, the signal was amplified by the numerous mDNAs released from the magnetic bead and the Exo III-assisted mDNA recycling. Under the optimal conditions, a detection limit down to 70 particles/μL was achieved, which is lower than the LODs of most currently available methods. Furthermore, this assay can be used to detect tumor exosomes in complex biological samples, demonstrating potential application in real sample diagnosis.
研究领域:
生物医学工程
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癌症诊断
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分子生物学
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BIOSENSORS & BIOELECTRONICS
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2019
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IF 9.518
Ruo-Nan Zhao,Li-Ping Jia,Zhe Feng,Rong-Na Ma,Wei Zhang,Lei Shang,Qing-Wang Xue,Huai-Sheng Wang
8-Hydroxy-2′-deoxyguanosine (8-OH-dG) is a principal stable marker of DNA oxidative damage . Sensitive and specific detection of 8-OH-dG is of great importance for early disease diagnosis. In this paper, we developed an electrochemiluminescence aptasensor for 8-OH-dG detection based on target induced multi-DNA release and nicking enzyme signaling amplification strategy. First, three kinds of short DNAs were aligned on the aptamers immobilized on the magnetic beads. In the presence of 8-OH-dG, the aptamer recognized and specifically bound with 8-OH-dG, leading to the release of three kinds of short DNAs and three-fold signal amplification . Then the released short DNAs hybridized with ferrocence (Fc) labeled hairpin DNA (Fc-HP) immobilized on the gold electrode to form a double strand DNA. Subsequently, nicking endonuclease (Nt. Alw I) recognized the asymmetric sequence in the dsDNA and cleaved the substrate strand (Fc-HP) into two parts, one fragments containing Fc would leave the surface of electrode. Based on the quenching effect of Fc on the electrochemiluminescence (ECL) of Ru(bpy) 3 2+ /TPA, a signal-on ECL aptasensor was developed. At the same time, three kinds of short DNAs were released again and reused to initiate the repeated cycles of hybridization-cleavage. Under double signal amplification, this aptasensor achieved a low detection of 25 fM and a wide linear range from 100 fM to 10 nM for 8-OH-dG. Besides, the amount of 8-OH-dG in urine samples derived from different people were determined with satisfactory results.
研究领域:
生物化学
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分析化学
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医学诊断
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高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2025
·
IF 5.4
Wushi Chen,Yunqin Shen,Hongyun Jia,Shanshan Hou,Lingjie Li,Lixia Huang,Jiyi Cai,Yuhan Xing,Yanqiu Chen,Yan Yang
Ethnopharmacological relevance Humulus lupulus L. , an herb traditionally employed in medicinal practices across North America, Egypt, India, and China, demonstrates broad therapeutic properties, including anti-inflammatory, antioxidant, hepatoprotective, and diuretic effects. Xanthohumol (XN), a prenylated flavonoid derived from Humulus lupulus L., retains these anti-inflammatory and antioxidant activities. However, its potential efficacy in allergic asthma remains largely unexplored. Aim of the study This study aimed to investigate the protective properties of XN against allergic asthma and to elucidate the underlying mechanisms, particularly the role of Absent in Melanoma 2 (AIM2), an innate immune sensor, in allergic asthma and its modulation by XN. Materials and methods The OVA-induced murine model of allergic asthma with XN administration in different dosages was constructed to evaluate the regulatory effects of XN on allergic asthma, particularly during the period of acute attack. Pathological alterations, T-cell polarization, inflammatory cytokine profiles, and oxidative stress markers were systematically assessed. Bioinformatic analysis, clinical case-control, and molecular docking combined with cellular thermal shift assay (CETSA) were applied to identify the specific target of XN to execute its effects. Human bronchial epithelial (16HBE) cell models with various stimulations and treatments were used in this study to verify the mechanistic exploration. Results In allergic asthmatic mice, the results demonstrated that XN dose-dependently attenuated airway inflammation and immune balance, characterized by reduced inflammatory cell infiltration, cytokine production, IgE secretion, and Th2 immune response. AIM2 was recognized as a critical binding target of xanthohumol through database analysis, molecular docking, and CETSA. Clinically, elevated serum AIM2 levels in asthma patients correlated positively with eosinophil percentage, IgE levels, and systemic immune inflammation index (SII). Mechanistically, both in vivo and in vitro studies revealed that XN mitigated oxidative stress and mitochondrial dysfunction, thereby reducing double-stranded DNA release, a key activator of the AIM2 inflammasome. Moreover, XN dose-dependently suppressed AIM2 inflammasome activation, an effect further validated in Poly (dA:dT)-stimulated and AIM2-silenced 16HBE cells. Conclusion This study provides the first evidence implicating XN can improve OVA-induced allergic asthma by suppressing AIM2 inflammasome activation, highlighting XN as a promising therapeutic candidate for allergic asthma.
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INTERNATIONAL JOURNAL OF PHARMACEUTICS
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2018
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IF 3.862
Qianhao Xu,Tianyuan Zhang,Qiyue Wang,Xinchi Jiang,Ai Li,Ying Li,Ting Huang,Fangyuan Li,Ying Hu,Daishun Ling,Jianqing Gao
Recent advances in nanomaterials have made iron oxide nanoparticles (IONPs) as an innovative approach for the delivery of genes. However, the effectiveness of IONPs-assisted gene delivery is currently suffering from their poor uniformity, which not only exhibits detrimental effect on the magnetic property, but also leads to the poor reproducibility to maintain the optimal gene delivery. To this end, the present study developed extremely uniform 15 nm-sized IONPs with a good monodispersity in water phase. These ultra-uniform IONPs exhibit notable potentials for an efficient gene delivery to human mesenchymal stem cells (hMSCs) with the assistance of magnetic force, which partly owes to their abilities to facilitate the cellular uptake, as well as to induce the rapid DNA release and the following nuclear transport. Besides, no significant detrimental effects of these IONPs are observed either on the proliferation or the multi-lineage differentiation potential of hMSCs. The current study highlights the potential advantages of designing extremely uniform magnetic nanomaterials for an efficient and safe delivery of genes to stem cells.
研究领域:
纳米技术
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生物材料
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干细胞生物学
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基因治疗
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Journal of Materials Chemistry B
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2013
·Vol. 1
·期 46
·6418-6426
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IF 0
Wenyu Li,Peng Zhang,Kun Zheng,Qiaoling Hu,Youxiang Wang
Extracellular stability to protect DNA against nucleases and stimulus-triggered intracellular DNA release are key factors in designing non-viral gene vectors. In this study, the diselenide-linked polycation mPEG–SeSe–PEI was developed as a new type of PEG-detachable gene vector for redox-responsive gene delivery. The corresponding stable analog mPEG–PEI and the disulfide-linked polycation mPEG–SS–PEI were synthesized as controls. The results showed that all the PEGylated polycations could condense DNA into tightly packed spherical nanoparticles about 80 nm in size, which showed excellent stability under physiological conditions. The results of zeta-potential measurements, stability tests and DNA release ability assay indicated that at a GSH concentration of 0.3 mM, the diselenide bonds were more easily cleaved than disulfide bonds, which facilitated dePEGylation and DNA release. Meanwhile, it was interestingly found that mPEG–SeSe–PEI/DNA polyplexes showed higher gene expression than mPEG–SS–PEI/DNA polyplexes in both HEK293T and HepG2 cells. Confocal laser scanning microscope (CLSM) images revealed that mPEG–SeSe–PEI/DNA polyplexes showed more efficient endosomal escape ability than mPEG–SS–PEI/DNA polyplexes. Based on these results, the diselenide bonds as a novel strategy are more suitable to address the challenging problem of extracellular stability and intracellular DNA release.
研究领域:
基因递送
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聚合物化学
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分子生物学
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生物医学工程
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PubMed
BIOMACROMOLECULES
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2019
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IF 5.667
Yuxin Zhang,Jie Zhou,Shengnan Ma,Yiyan He,Jun Yang,Zhongwei Gu
Smart polymers as ideal gene carriers have drawn increasing attentions due to the effective DNA release once triggered by intrinsic stimuli, as well as reduced cytotoxicity. Herein, a stimulus-responsive, positively charged and water-soluble polymer (OEI-TKx) was facilely engineered by cross-linking low molecular weight oligoethylenimine (OEI) via thioketal (TK) linkages that would cleave selectively in reactive oxygen species (ROS)-rich environments induced by hypoxia. Agarose gel electrophoresis assay demonstrated that the threshold N/P ratios for complete retardation of negatively charged DNA migration were above 5 for OEI-TKx. The reduction in DNA-condensing capability and the changes in particle size, size distribution and particle morphology all illustrated that OEI-TKx possessed excellent ROS responsiveness. OEI-TKx/DNA polyplexes showed lower toxicity and higher gene transfection efficiency compared with PEI/DNA polyplexes. The optimum formulation, OEI-TKx/DNA polyplexes (N/P = 40), showed a little better performance than PEI/DNA polyplexes in cellular uptake profile. Furthermore, OEI-TKx/DNA polyplexes could escape from endosomes to the cytosol as efficiently as PEI/DNA polyplexes. Confocal images confirmed that OEI-TKx/DNA polyplexes could more effectively release DNA than PEI/DNA polyplexes, mainly owing to the valid cleavage of thioketal linkages induced by characteristic rich-ROS in Hela cells. These results suggested that OEI-TKx could represent an on-demand stimulus-responsive gene delivery platform.
研究领域:
生物医学工程
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药学
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纳米技术
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分子生物学
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癌症治疗
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高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2024
·
IF 18.5
Xueqin Zhu,Xiaoxi Wang,Zimai Liu,Bing Jiang,Zonghong He,Sijia Liu,Yahong Wu,Zixian Wu,Tiantian Zhang,Meiyi Liu,Kai Li,Xiaoshuang Niu,Yanfeng Gao
Although numerous peroxidase (POD)-like nanozymes have been designed for catalytic therapy of cancer, development of nanozymes with higher therapeutic efficacy and less adverse effects are challengeable. More importantly, the underlying antitumor mechanism remains largely unknown which hinders their application. Here, the nitrogen-doped carbon nanozyme (N-PCNS) as a model is utilized and demonstrated that its capacity to specifically activate the STING pathway in tumor cells through reactive oxgen species (ROS)-mediated mitochondrial DNA (mtDNA) release, which provides the initial signals for STING-dependent innate immune response. Further, a peptide-nanozyme conjugate (PNEC, OPBP1-N-PCNS), comprising a PD-L1 blocking/targeting dual-functional peptide and a cationic-coated N-PCNS is constructed, which are conjugated through a matrix metalloproteinase responsive peptide linker. The cleaved derivate nanozyme with positive charge exhibits recyclable capacity and superior ability in capturing and enhancing the transcellular transport of tumor-derived mtDNA toward DCs, thereby amplifying the STING signaling-mediated anti-tumor immune response. This study proposed a unique mechanism and design strategy for POD-like nanozyme in cancer immunotherapy.
研究领域:
癌症免疫治疗
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纳米技术
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免疫学
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分子生物学
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高影响因子
CARBOHYDRATE POLYMERS
·
2024
·
IF 10.7
Jing Feng,Yiting Liu,Xiaoran Zheng,Min Gao,Li Wang,Lígia R. Rodrigues,Yuting Wen,Hangcheng Pan,Gege Li,Longjiang Zhang,Bing Wan,Yunlei Zhang
The efficacy of cancer therapies is significantly compromised by the immunosuppressive tumor milieu. Herein, we introduce a previously unidentified therapeutic strategy that harnesses the synergistic potential of chitosan-coated bacterial vesicles and a targeted chemotherapeutic agent to activate dendritic cells, thereby reshaping the immunosuppressive milieu for enhanced cancer therapy. Our study focuses on the protein-mediated modification of bacterium-derived minicells with chitosan molecules, facilitating the precise delivery of Doxorubicin to tumor sites guided by folate-mediated homing cues. These engineered minicells demonstrate remarkable specificity in targeting lung carcinomas, triggering immunogenic cell death and releasing tumor antigens and damage-associated molecular patterns, including calreticulin and high mobility group box 1. Additionally, the chitosan coating, coupled with bacterial DNA from the minicells, initiates the generation of reactive oxygen species and mitochondrial DNA release. These orchestrated events culminate in dendritic cell maturation via activation of the stimulator of interferon genes signaling pathway, resulting in the recruitment of CD4 + and CD8 + cytotoxic T cells and the secretion of interferon-β, interferon-γ, and interleukin-12. Consequently, this integrated approach disrupts the immunosuppressive tumor microenvironment, impeding tumor progression. By leveraging bacterial vesicles as potent dendritic cell activators, our strategy presents a promising paradigm for synergistic cancer treatment, seamlessly integrating chemotherapy and immunotherapy.
研究领域:
癌症治疗
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免疫疗法
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生物医学工程
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药理学
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分子生物学
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细胞生物学
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高影响因子
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