RSC Advances
·
2025
·Vol. 15
·期 54
·46266-46276
·
IF 4.6
Yiyan Song,Dawei Yu,Zhijie Sheng,Huafeng Song,Junchi Xu,Jianping Zhang,Hui Chen,Ping Xu
Tuberculosis, the world's leading cause of death from a single infectious agent in 2023, is exacerbated by the rising prevalence of multidrug-resistant (MDR) Mycobacterium tuberculosis, underscoring an urgent need for novel therapeutic agents. In this study, we developed a green in situ synthesis strategy for silver nanoparticles (Ag NPs) utilizing polydopamine (PDA) as a biomimetic reducing and coating agent on mesoporous silica (SBA-15). The material was further co-coated with the antimicrobial polymer epsilon-poly-L-lysine (EPL) and functionalized with mannose via a Schiff-base reaction. The resulting composite, denoted as Ag@SBA-15/Man-PDA-EPL, was comprehensively characterized by physicochemical techniques and exhibited low cytotoxicity. Mycobacterium tuberculosis (H37Rv and MDR) was used for the evaluation of the prepared nanocomposites in which the components (Ag@SBA-15/PDA and EPL) exhibited synergistic effect towards MDR. The mannose-grafted composite (Ag@SBA-15/Man-PDA-EPL) exhibited significantly enhanced anti-tubercular efficacy compared to its non-functionalized counterpart (Ag@SBA-15/PDA-EPL), an improvement attributed to the antibacterial capability conferred by the Schiff-base formation. Transmission electron microscopy (TEM) imaging confirmed extensive cellular damage in mycobacteria treated with Ag@SBA-15/Man-PDA-EPL. This study validates a rational combination strategy integrating silver nanoparticles, EPL, and ligand-specific targeting via a Schiff-base moiety into a unified nanoplatform, offering a promising approach to combat drug-resistant tuberculosis and mitigate nosocomial infections.
研究领域:
纳米医学
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抗菌治疗
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材料科学
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传染病学
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药物递送
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MICROBIOLOGY
·
2024
·Vol. 62
·期 1
·49-62
·
IF 3.3
Xu Tao,Wang Chutong,Li Minying,Wei Jing,He Zixuan,Qian Zhongqing,Wang Xiaojing,Wang Hongtao
Tuberculosis (TB), a bacterial infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis ), is a significant global public health problem. Mycobacterium tuberculosis expresses a unique family of PE_PGRS proteins that have been implicated in pathogenesis. Despite numerous studies, the functions of most PE_PGRS proteins in the pathogenesis of mycobacterium infections remain unclear. PE_PGRS45 (Rv2615c) is only found in pathogenic mycobacteria. In this study, we successfully constructed a recombinant Mycobacterium smegmatis ( M. smegmatis ) strain which heterologously expresses the PE_PGRS45 protein. We found that overexpression of this cell wall-associated protein enhanced bacterial viability under stress in vitro and cell survival in macrophages. MS_PE_PGRS45 decreased the secretion of pro-inflammatory cytokines such as IL-1β, IL-6, IL-12p40, and TNF-α. We also found that MS_PE_PGRS45 increased the expression of the anti-inflammatory cytokine IL-10 and altered macrophage-mediated immune responses. Furthermore, PE_PGRS45 enhanced the survival rate of M. smegmatis in macrophages by inhibiting cell apoptosis. Collectively, our findings show that PE_PGRS45 is a virulent factor actively involved in the interaction with the host macrophage.
研究领域:
微生物学
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免疫学
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传染病学
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DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2016
·
IF 2.243
Xiaoxia Shi,Shanshan Sha,Likun Liu,Xin Li,Yufang Ma
Mycobacterium tuberculosis dTDP- d -glucose 4,6-dehydratase (RmlB) is the second enzyme for the biosynthesis of dTDP- l -rhamnose, which is a sugar donor to the synthesis of the cell wall linker, d - N -acetylglucosamine- l -rhamnose. RmlB is essential to mycobacterial growth and is not found in humans; therefore, it is a potential target for developing new anti-tuberculosis drugs. So far, there has been no suitable method for high-throughput screening of RmlB inhibitors. Here, the recombinant M. tuberculosis RmlB was purified and an absorbance-based microtiter plate assay was developed for RmlB activity. It could be used for high-throughput screening of RmlB inhibitors. The kinetic properties of M. tuberculosis RmlB, including optimal pH, optimal temperature, the effect of metal ions , and the kinetic parameters , were determined with this assay. The inhibitory effects of dTTP and dTDP on M. tuberculosis RmlB were also studied with the assay.
研究领域:
微生物学
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药物开发
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生物化学
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DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2020
·
IF 7.632
Xia Xiao,Jing-Kai He,Yi-Xin Guan,Shan-Jing Yao
Androst-4-ene-3,17-dione (AD) is an important intermediate of steroid drugs, and the microbial side-chain cleavage of sterols to produce AD by Mycobacterium sp. has attracted much attention. Unfortunately, a low yield of this reaction due to the extremely poor solubility of sterols in aqueous medium is a critical problem. Ionic liquid (IL) as a promising alternative to organic solvents is deemed to increase the solubility of hydrophobic substrates; however, the tolerance of microorganisms to ILs is limited and biocompatible ILs are desperately needed. In this work, cholinium amino acids ([Ch][AA]) ILs were first introduced as cosolvents for the whole-cell bioconversion of phytosterols to produce AD. The physicochemical properties of these novel ILs were investigated on the basis of their structures, and the solubility of phytosterols and AD in [Ch][AA] ILs aqueous solution was determined. The results indicated that moderate cell membrane permeability was obtained by [Ch][AA] ILs, and [Ch][Asp], [Ch][Glu], and [Ch][Gln] showed proper biocompatibility to Mycobacterium sp. MB 3683 cells among 18 kinds of [Ch][AA] ILs. The highest production of AD was achieved in 1% (v/v) [Ch][Asp], which was about twice as high as that in the buffer. Therefore, [Ch][AA] ILs are potential cosolvents in steroid biotransformation to promote medium engineering for a sustainable environment.
研究领域:
生物化学
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环境科学
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微生物学
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DOI
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2019
·
IF 2.705
Fake Li,Jie Luo,Huan Xu,Yang Wang,Wenbin Jiang,Kai Chang,Shaoli Deng,Ming Chen
Early secreted antigenic target 6-kDa protein (ESAT6) is an essential virulence factor of Mycobacterium tuberculosis (MTb). However, ESAT6 helped fighting MTb infection according to vaccine studies. It's unclear whether ESAT6 confers protection via enhancing the innate immunity of macrophages, which are the first-line defense against MTb. We profiled the global transcriptional changes and characterized the innate immunity of THP-1 macrophages treated with ESAT6. We found ESAT6 promoted the phagocytosis ability, enhanced reactive oxygen species (ROS) generation and accelerated glucose metabolism in macrophages. Meanwhile, ESAT6 induced a distinctive phenotype of macrophages with a concurrence of pro-inflammatory and anti-inflammatory cytokines and chemokines . ESAT6 increased the expression of HIF1α mRNA and protein. Interfering HIF1α with siRNA defected the capacity of phagocytosis and ROS generation as well as glucose metabolism. Thus, ESAT6 enhanced the protective innate immunity of macrophages partially via HIF1α. This study provided clues for developing therapies against tuberculosis by targeting ESAT6.
研究领域:
免疫学
·
微生物学
·
传染病学
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DOI
PubMed
高影响因子
Vaccines
·
2024
·Vol. 12
·期 6
·622
·
IF 5.2
Shufeng Weng,Qingchun Li,Tianran Zhang,Taiyue Lin,Yumo He,Guang Yang,Honghai Wang,Ying Xu
Tuberculosis (TB) is a major global health threat despite its virtual elimination in developed countries. Issues such as drug accessibility, emergence of multidrug-resistant strains, and limitations of the current BCG vaccine highlight the urgent need for more effective TB control measures. This study constructed BCG strains overexpressing Rv1002c and found that the rBCG-Rv1002c strain secreted more glycosylated proteins, significantly enhancing macrophage activation and immune protection againstMycobacterium tuberculosis(M. tb). These results indicate that Rv1002c overexpression promotes elevated levels of O-glycosylation in BCG bacteriophages, enhancing their phagocytic and antigenic presentation functions. Moreover, rBCG-Rv1002c significantly upregulated immune regulatory molecules on the macrophage surface, activated the NF-κB pathway, and facilitated the release of large amounts of NO and H2O2, thereby enhancing bacterial control. In mice, rBCG-Rv1002c immunization induced greater innate and adaptive immune responses, including increased production of multifunctional and long-term memory T cells. Furthermore, rBCG-Rv1002c-immunized mice exhibited reduced lung bacterial load and histological damage uponM. tbinfection. This result shows that it has the potential to be an excellent candidate for a preventive vaccine against TB.
研究领域:
免疫学
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微生物学
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疫苗学
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传染病
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2026
·
IF 2.5
Yingxiao Han,Songwei Ma,Rui Ma,Jingjing Li,Lei Qiu,Jia wen Ma,Yanyan Ma,Mengyin Tang,Zhijuan Cao,Ziqi Li,Shulin zhang,Guorong Ma
Mycobacterium tuberculosis culture filtrate proteins (MTB-CFPs) are critical for tuberculosis diagnosis and vaccine development. To explore their diagnostic potential, MTB was statically cultured in Sauton's medium for 15 days; MTB-CFPs were collected, fractionated using six biomimetic affinity chromatography (BiAC) resins, and analyzed by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Four MTB-specific regions of difference (RDs) and the PstS1 protein were recombinantly expressed and purified, with their serological efficacy assessed by ELISA using sera from 157 pulmonary tuberculosis (pTB) patients and 71 healthy controls (HCs). LC-MS/MS identified 622 MTB proteins, 307 of which were validated with ≥2 peptides and P < 0.05. BiAC fractionation significantly increased the average peptide number (2.09-fold), peptide coverage (1.43-fold), and the exponentially modified Protein Abundance Index (emPAI) (1.70-fold) in 281 common identifications. Receiver Operating Characteristic (ROC) analysis revealed rEC (a fusion expression product of the Rv3875 and Rv3874 genes in the RD1 region, linked by a 15-nucleotide connector) as the optimal diagnostic marker Area Under the Curve (AUC = 0.8030), with sharply distinct IgG levels between pTB patients and HCs ( P < 0.0001). Notably, rEC combined with GmdA (encoded by the Rv1511 gene located in RD11) enhanced sensitivity while preserving high specificity, especially for Acid-Fast Bacilli (AFB) negative pTB cases demonstrating substantial clinical translation potential. Additionally, extended culture promoted MTB autolysis and cytoplasmic protein release, providing valuable insights into TB pathogenesis and biomarker discovery. This integrated approach of BiAC fractionation, proteomics, and serological validation advances TB precision diagnostics.
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DOI
PubMed
高影响因子
Frontiers in Chemistry
·
2025
·
IF 4.2
Ting Yuan,Jianbo Yuan,Jian Huang,Nana Li,Huan Cai,Yan Yang,Juan Li,Rui Chen,Xun Min
Introduction: The rapid and accurate identification of Mycobacterium tuberculosis (MTB) is essential for effective tuberculosis (TB) control. However, conventional diagnostic methods for MTB suffer from limitations such as low sensitivity, poor specificity, high cost, reliance on specialized instruments, and complex, time-consuming procedures. To address these challenges, there is an urgent need for a simple, rapid, and highly sensitive detection method that can be deployed in point-of-care settings.Methods: We developed a one-pot biosensing system combining recombinase polymerase amplification (RPA) and CRISPR/Cas12a-driven G4-hemin self-assembly for the colorimetric detection of MTB. Glycerol was employed as a phase-separation barrier to prevent interference between RPA amplification and CRISPR/Cas12a trans-cleavage. A single-stranded DNA (ssDNA) probe, designed to self-assemble with ssDNA-hemin into G4-hemin nanozymes upon CRISPR/Cas12a-mediated cleavage, served as the reaction substrate. The ssDNA-hemin further enhanced the catalytic activity of the generated G4-hemin DNAzyme. The entire assay was completed in a single step within 60 min without requiring complex instrumentation.Results and Discussion: Under optimized conditions, the biosensing system achieved ultrasensitive naked-eye detection of MTB with a limit of detection (LOD) of 10 copies/μL, comparable to traditional four-step fluorescent assays. Clinical validation using 104 patient samples demonstrated high concordance with standard diagnostic methods. This approach combines the advantages of recombinase polymerase amplification (RPA), CRISPR/Cas12a specificity, and G4-hemin DNAzyme-based colorimetric signal amplification, enabling simple, equipment-free visual detection. Given its speed, sensitivity, and ease of use, this biosensing system holds significant promise for point-of-care MTB nucleic acid testing in resource-limited settings.
研究领域:
生物传感器
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分子诊断
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CRISPR技术
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核酸扩增
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DNA酶技术
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结核病检测
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2024
·
IF 2.7
Qiong Liu,Mei Yang,He Zhang,Wenjie Ma,Xin Fu,Huiqing Li,Sainan Gao
Tuberculosis caused by Mycobacterium bovis poses a global infectious threat to humans and animals. Therefore, there is an urgent need to develop a sensitive, precise, and easy-to-readout strategy. Here, a novel tandem combination of CRISPR/Cas12a system with dual HCR (denoted as CRISPR/Cas12a-D-HCR) was constructed for detecting Mycobacterium bovis. Based on the efficient trans-cleavage activity of the active CRISPR/Cas12a system, tandem-dsDNA with PAM sites was established using two flexible hairpins, providing multiple binding sites with CRISPR/Cas12a for further amplification. Furthermore, the activation of Cas12a initiated the second hybridization chain reaction (HCR), which integrated complete G-quadruplex sequences to assemble the hemin/G-quadruplex DNAzyme. With the addition of H2O2 and ABTS, a colorimetric signal readout strategy was achieved. Consequently, the CRISPR/Cas12a-D-HCR achieved a satisfactory detection linear range from 20 aM to 50 fM, and the limit of detection was as low as 2.75 aM with single mismatched recognition capability, demonstrating good discrimination from different bacterial species. Notably, the practical application performance via the standard addition method was verified, with the recovery ranging from 96.0% to 105.2% and the relative standard deviations (RSD) ranging from 0.95% to 6.45%. The proposed CRISPR/Cas12a-D-HCR sensing system served as a promising application for accurate detection in food safety and agricultural fields.
研究领域:
分子生物学
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病原微生物学
·
食品安全检测
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产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2015
·
IF 3.989
Deng Wanyan
PE/PPE family proteins, named after their conserved PE (Pro-Glu) and PPE (Pro-Pro-Glu) domains of N-terminal, are most intriguing aspects of pathologic mycobacterial genome. The roles of most members of this family remain unknown, although selected genes of this family are related to the virulence of Mycobacterium tuberculosis. In order to decipher the role of Rv1169c, the Mycobacterium smegmatis strain heterologous expressed this ORF was constructed and identified that Rv1169c was a cell wall associated protein with a novel function in modifying the cell wall fatty acids. The growth of Rv1169c expressing strain was affected under surface stress, acidic condition and antibiotics treatment. M. smegmatis expressing Rv1169c induced necrotic cell death of macrophage after infection and significantly decreased interlukin-6 production compared to controls. In general, these results underscore a proposing role of Rv1169c in virulence of M. tuberculosis, as it's role in the susceptibility of anti-mycobacteria factors caused by modified cell wall fatty acid, and the induced necrotic cell death by Rv1169c is crucial for M. tuberculosis virulence during infection.
研究领域:
微生物学
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免疫学
·
分子生物学
·
传染病学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2018
·
IF 3.118
Shuangquan Yan,Junfeng Zhen,Yue Li,Chenhui Zhang,Andrea Stojkoska,Nzungize Lambert,Qiming Li,Ping Li,Jianping Xie
The success of Mycobacterium tuberculosis as a pathogen largely contributes to its ability to infect, modify and persist within the host cells. M. tuberculosis Rv0177 is a gene of the mce1 operon (Mammalian cell entry), encoding a conserved hypothetical protein, essential for M. tuberculosis survival and up-regulated within murine macrophages. To explore its function, Rv0177 was heterologously expressed in M. smegmatis . The recombinant protein was located in the cell wall. M. smegmatis recombinant strain expressing Rv0177 altered sliding motility, its cell wall architecture and the permeability. Moreover, M. smegmatis expressing Rv0177 could up-regulate MCP-1 and downregulate the IL-6 expression in RAW264.7 macrophages in comparison to the control. MS_Rv0177 increased the expression of MCP-1 inducible protein (MCPIP) and a C/EBP homologous protein (CHOP) owing to MCP-1. In addition, the JNK signaling pathway was engaged in the interplay between MS_Rv0177 and macrophages. The macrophage caspase-3 activation and cell apoptosis were induced by the recombinant. This provided novel functional cues for the MCE-associated Rv0177.
研究领域:
微生物学
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免疫学
·
分子生物学
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DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Yishi Li,Dengyong Peng,Shuliang Guo,Bijun Yang,Jing Zhou,Jiaxu Zhou,Qifan Zhang,Lijuan Bai
Rapid and sensitive tuberculosis (TB) diagnoses remain big challenges to current detection tools. In this work, a sensitive electrochemical aptasensor was constructed for the determination of Mycobacterium tuberculosis antigen MPT64 using a new redox nanoprobe. We found that anthraquinone derivative, anthraquinone-2-carboxylic acid (AQCA), a redox mediator, could be confined in ordered mesoporous carbon material of CMK-3. Due to the large loading amount of AQCA , as well as the confined space and electron transfer promotion effect of CMK-3, the obtained AQCA/CMK-3 nanohybrid with mass ratio of 2:1 showed excellent electroactivity and was employed as a new redox nanoprobe for signal amplification for the first time. Additionally, urchin-like Ce-MOFs were used to load a large amount of deposited gold nanocrystals (dep-Au), leading to dense immobilization of capture probe. The proposed electrochemical aptasensor for MPT64 detection showed a good linear relationship in the range from 100 fg/mL to 10 ng/mL with a low detection limit of 67.6 fg/mL. Besides, the aptasensor was utilized to detect MTP64 in human serum samples for TB diagnosis and presented satisfactory results.
研究领域:
生物医学工程
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电化学
·
纳米技术
·
诊断学
·
分析化学
✓ 已验证
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）