ACS Applied Materials & Interfaces
·
2016
·
IF 7.145
Shiwei Zhou,Yingying Wang,Jun-Jie Zhu
B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) are often used to monitor the apoptosis of tumor cells and evaluate cancer drug effect. In this work, a novel sandwich-type dual-signal-marked electrochemical biosensor was fabricated for simultaneous detection of Bcl-2 and Bax proteins. Reduced graphene oxide (RGO) layers were used as substrate to immobilize Bcl-2 and Bax antibodies for further capturing target antigens. CdSeTe@CdS quantum dots (QDs) and Ag nanoclusters (NCs) with antibody modification and mesoporous silica amplification were used as signal probes, which were proportional to the amount of Bcl-2 and Bax antigens. Mesoporous SiO2 can provide a larger surface area, more effectively charged by ethylene imine polymer or poly(diallyldimethylammonium chloride) to adsorb more probes. The Bcl-2 and Bax proteins were determined indirectly by the detection of oxidation peak currents of Cd and Ag using anodic stripping voltammetry, showing a good linear relationship in the protein concentration range from 1 ng/mL to 250 ng/mL. The detection limit of trace protein level was ∼0.5 fmol. The biosensor was further introduced to investigate Bcl-2 and Bax expressions from nilotinib-treated chronic myeloid leukemia K562 cells. With the increase of drug dosage and incubation time, the up-regulation for Bax and down-regulation for Bcl-2 were observed, which indicated that the apoptosis level of K562 cells could be regulated by Bcl-2 family. The ratio of Bax/Bcl-2 was further calculated for evaluation of its drug effect and apoptosis level. The limited cell amount for detection reached less than 1 × 103 cells, much lower than traditional methods. Furthermore, completely independent detection step and stable acid solutions containing Ag+ and Cd2+ for long-time storage contribute to reducing the error from the sample differences and avoiding the potential errors from the photodegradation of fluorescent probes, enzymolysis of DNA, or inactivation of enzyme during an excess experimental period.
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Molecular Catalysis
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2022
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IF 5.089
Ying Zhang,Zhi-Wen Duan,Han-Yu Liu,Feng Qian,Pu Wang
Microbial whole-cell catalysis has been a popular method for chiral alcohols preparation. In this study, Cyberlindnera saturnus ZJPH1807 was successfully employed as a biocatalyst to reduce 2-chloro-1-(2,4-dichlorophenyl)ethanone (CPE) to ( R )-2-chloro-1-(2,4-dichlorophenyl)ethanol (( R )-CPEO), a key chiral intermediate of antifungal drug miconazole. In order to further improve the bioprocess yield, natural deep eutectic solvent and cyclodextrin were introduced into the reaction system as additives. It was found that carnitine/trehalose (C/Tre, 1:1 mol/mol) could increase the permeability of cell membrane appropriately and improve the mass transfer rate. And the addition of methyl-β-cyclodextrin (MCD) could improve the solubility of CPE in buffer solution, simultaneously promoting cofactor recycling. Furthermore, MCD and C/Tre exerted a synergistic effect on the acceleration of CPE bioreduction, producing 91.0% yield in 1.5 h at 35.8 mM CPE in a C/Tre-MCD-containing system, with a 12.4-fold increase in productivity compared to neat buffer solution only. When the substrate loading was increased to 67.1 mM, the yield could also achieve 84.2%. The established bioreduction process was also feasible at 100 mL preparation scale with a 88.5% ( R )-CPEO yield. The integration of C/Tre and MCD can significantly improve catalytic efficiency for hydrophobic substrates and be applied in biocatalysis.
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MICROCHEMICAL JOURNAL
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2020
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IF 3.594
Bokai Ma,Fujian Xu,Mian He,Yuqing Lin,Guanghui Hu,Mei Zhang,Xinying Zhao,Weili Liu
Rhodamine B hydrazide (RBH) was prepared as a new derivatizing reagent for residual formaldehyde in a drug such as N-butyl-2-cyanoacrylate (NBCA). RBH reacted with formaldehyde rapidly at room temperature, and the resulting derivative can be detected sensitively at 238 nm. After optimizing the derivatization and chromatographic separation conditions, RBH as a pre-column derivatizing reagent has been successfully used for the analysis of formaldehyde in real NBCA samples by high-performance liquid chromatography (HPLC). The detection limit of the method is 0.004 μg/mL for formaldehyde, and the recovery is 94.0%-96.6%. Most notably, compared with the previous HPLC methods, the developed method has the advantages of mild derivatization temperature, short derivatization time and high sensitivity.
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BIOORGANIC CHEMISTRY
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2021
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IF 5.275
Xinjian Yin,Litong Chen,Siwen Yuan,Lan Liu,Zhizeng Gao
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC 50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC 50 (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z’ factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.
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Catalysts
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2019
·Vol. 9
·期 6
·526
·
IF 3.444
Ran Cang,Li-Qun Shen,Guang Yang,Zhi-Dong Zhang,He Huang,Zhi-Gang Zhang
Value-added utilization of biomass-derived 5-hydroxymethylfurfural (HMF) to produce useful derivatives is of great interest. In this work, extremely radiation resistantDeinococcus wulumuqiensisR12 was explored for the first time as a new robust biocatalyst for selective oxidation of HMF to 5-hydroxymethylfuroic acid (HMFCA). Its resting cells exhibited excellent catalytic performance in a broad range of pH and temperature values, and extremely high tolerance to HMF and the HMFCA product. An excellent yield of HMFCA (up to 90%) was achieved when the substrate concentration was set to 300 mM under the optimized reaction conditions. In addition, 511 mM of product was obtained within 20 h by employing a fed-batch strategy, affording a productivity of 44 g/L per day. Of significant synthetic interest was the finding that theD. wulumuqiensisR12 cells were able to catalyze the selective oxidation of other structurally diverse aldehydes to their corresponding acids with good yield and high selectivity, indicating broad substrate scope and potential widespread applications in biotechnology and organic chemistry.Keywords:biocatalysis;extremophile;5-hydroxymethylfurfural;5-hydroxymethylfuroic acid;platform chemicals;whole cells
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Oxidative Medicine and Cellular Longevity
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2019
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IF 4.868
Yang Zhao,Yan Cai,Li-Yan Cui,Wen Tang,Bo Liu,Jia-Jia Zheng,Wen-Zhe Si,Xian Wang,Ming-Jiang Xu
Aims. Vascular calcification (VC) is a primary risk factor for cardiovascular mortality in chronic renal failure (CRF) patients; thus, effective therapeutic targets are urgently needed to be explored. Here, we identified the role of intestinal bacterial translocation in CRF-related VC. Methods and Results. Antibiotic supplementation by oral gavage significantly suppressed intestinal bacterial translocation, CRF-related VC, and aortic osteogenic gene and Toll-like receptor (TLR) gene expression in CRF rats. Furthermore, TLR4 and TLR9 activation in vascular smooth muscle cells (VSMCs) aggravated inorganic phosphate- (Pi-) induced calcification. TLR9 inhibition, but not TLR4 inhibition, by both a pharmacological inhibitor and genetic methods could significantly reduce CRF rats’ serum or CRF-induced VC. Interestingly, bone morphogenic protein-2 (BMP-2) levels were increased in the aorta and sera from CRF rats. Increased BMP-2 levels were also observed in VSMCs treated with TLR9 agonist, which was blocked by NF-κB inhibition. Both siRNA knockdown of BMP-2 and NF-κB inhibitor significantly blocked TLR9 agonist-induced VSMC calcification. Conclusions. Gut bacterial translocation inhibited by oral antibiotic significantly reduces CRF-related VC through inhibition of TLR9/NF-κB/BMP-2 signaling.
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APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2022
·Vol. 194
·期 11
·5274-5291
·
IF 3.094
Lei Jiping,Tang Kexin,Zhang Ting,Li Yan,Gao Zhen,Jia Honghua
2-O-α-D-Glucosyl glycerol (2-αGG) can be used as a multipurpose anti-aging, cell-stimulating, and skin moisturizing agent in the cosmetic industry. Sucrose phosphorylase (SPase) has been widely used in the production of 2-αGG. In this paper, the gene encoding sucrose phosphorylase from Bifidobacterium longum (BlSP) was inserted into pRSF-Duet-1 to construct the recombinant plasmid pRSF-BlSP and was functionally expressed in E. coli BL21(DE3) to be used as a biocatalyst for the synthesis of 2-αGG firstly. The mutations of BlSP were carried out based on alanine scanning, and a positive mutant G293A with a 50% increase in activity for 2-αGG production was identified. Mutant G293A has less K m and bigger k cat / K m towards glycerol than the parental BlSP. Subsequently, the production of 177.6 g/L 2-αGG was attained from 1 M sucrose and 1.2 M glycerol catalyzed by 17 mg/mL G293A mutant. This study indicated that BlSP has good potential in the production of 2-αGG.
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RENEWABLE ENERGY
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2021
·
IF 8.001
Hongyou Wan,Fuzhen Wang,Yuhan Chen,Zisheng Zhao,Guangyi Zhang,Ming Dou,Binghan Xue
In this study, nano-Fe 3 O 4 supporting on granular activated carbon (NFGAC) were prepared, characterized and used as an individual additive to promote the anaerobic digestion of azo dye Reactive Red 2 (RR2) wastewater. Results showed that organics removal and methane production were effectively promoted with the addition of NFGAC. Compared with the control reactor, the RR2 removal efficiency increased by 117.9% and the methane production increased by 167.2% in the 0.18-NFGAC added reactor when sole RR2 used as the substrates. Further analysis indicated that during the RR2 anaerobic digestion process the electron transfer system (ETS) activity was effectively improved with the addition of NFGAC. One of the possible mechanism was that the loading nano-Fe 3 O 4 on the surface of GAC could increase the number of functional groups in NFGAC, which were beneficial to enhancing the electron accepting capacities (EAC) and electron donating capacities (EDC). Microbial community analysis showed that the bacteria ( Halomonas , Pseudomonas and Bacillus ) relevant to RR2 removal were enriched in NFGAC added reactors, which played an important role during RR2 removal process. In addition, the addition of NFGAC could promote potential direct interspecies electron transfer (DIET) between Pseudomonas and Methanosarcina.
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CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Junxin Jia,Yaqi Jiao,Jiaying Gu,Chunnan Zhou,Qin Zhou,Hui Li,Sa Wang,Hongtao Shi,Li Li
Phenanthrene (PHE) is a typical polycyclic aromatic hydrocarbon (PAH) pollutant characterized by biological toxicity and environmental persistence. To enhance its degradation efficiency, a composite system combining birnessite with Novosphingobium sp. HDJX-2 was established to investigate the synergistic mechanism between minerals and functional bacteria. Gas chromatography-mass spectrometry (GC–MS), coupled with transcriptomic and metabolomic analyses, was employed to elucidate the molecular regulation and metabolic pathway reconstruction involved in PHE degradation in the composite system. The results demonstrated that PHE was metabolized by the strain through a series of reactions, including aromatic ring dioxygenation, ring cleavage, and decarboxylation, leading to the formation of acetyl-CoA via the phthalic acid pathway and ultimately achieving complete mineralization through the tricarboxylic acid (TCA) cycle. This degradation process was accompanied by extensive transcriptional regulation and metabolic pathway enrichment, with 967 differentially expressed genes and 125 significantly altered metabolites identified. Further analysis revealed the upregulation of genes associated with biofilm formation ( rcsA , 2.31-fold upregulation) and energy metabolism ( Rbs , 2.47-fold upregulation), along with enhanced pathways, such as arachidonic acid metabolism (1.85-fold upregulation) and riboflavin metabolism (1.13-fold upregulation). These changes contributed to improved membrane transport, oxidative stress response, and energy production, thereby significantly promoting PHE degradation. Notably, pathways related to amino acid metabolism and carbon fixation were significantly enriched, further enhancing the system's degradation capacity. This study established an efficient and stable mineral–microbial synergistic system and revealed its multilayered regulatory mechanisms in PAH bioremediation, offering theoretical insights and practical strategies for ecological restoration.
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Materials Today Chemistry
·
2025
·
IF 6.7
Ruoyu Di,Ruiyue Hang,Huanming Chen,Liwei Yang,Yuyu Zhao,Xiaohong Yao,Ruiqiang Hang
Infections and loosening remain two significant challenges in the clinical application of titanium (Ti) implants. Zinc oxide (ZnO) nanostructured coatings possess remarkable antibacterial properties and osteogenesis potential, but their instability-induced physiological toxicity needs to be improved. In this work, ZnO/zinc phosphate (Zn 3 (PO 4 ) 2 ) composite micro/nano-scaled flower-like structural coatings were constructed on Ti implant surfaces by magnetron sputtering combined with two-step hydrothermal treatments to address these concerns. The results showed that the composite micro/nano-structures became larger with the increase of the concentration of the second hydrothermal precursor (dipotassium hydrogen phosphate), while the coatings became more hydrophilic. In addition, the composite coatings exhibited excellent antibacterial properties, mediated macrophage polarization towards anti-inflammatory and pro-healing M2 phenotype, and promoted osteogenic differentiation of bone marrow mesenchymal stem cells. This surface modification strategy, which utilizes the partial conversion of ZnO to Zn 3 (PO 4 ) 2 to balance the antibacterial and cell-modulating properties of Ti implants, holds great promise for clinical applications.
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EXPERIMENTAL NEUROLOGY
·
2025
·
IF 4.2
Jun Fu,Chengjie Ding,Wenqing Li,Jiali Lei,He Wang,Lingfei Meng,Xiaoya Lin,Weiyuan Chen,Hongchang Gao
Alzheimer's Disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline, with hallmark pathological features including Aβ deposition and tau protein hyperphosphorylation. These pathological alterations are particularly prominent in the Prefrontal Cortex (PFC) and exert profound effects on cognitive function. The present study aimed to reveal key proteins in the PFC and provide insight into their potential mechanisms of action in AD. Accordingly, proteomic analysis was conducted to profile differential proteins in 5 × FAD mice, followed by in vivo and in vitro validation of core proteins through AAV-mediated interference of target gene expression. Furthermore, transcriptomics and molecular biology techniques were employed to confirm the underlying mechanisms of action of core proteins in AD. The results demonstrated that ELAVL1 was significantly upregulated in the PFC of AD mice. Behavioral experiments revealed that inhibition of ELAVL1 expression markedly improved long-term memory in AD mice. Immunostaining and Western blot analyses confirmed that suppressing ELAVL1 expression alleviated Aβ1–42 deposition and apoptosis. In vitro experiments indicated that knocking down ELAVL1 significantly reduced the apoptotic response of SH-SY5Y cells induced by Aβ1–42 and APP/Swe cells. Transcriptomic analysis further revealed that ELAVL1 regulates Aβ1–42-induced apoptosis via activation of the Bcl-2/Bax pathway. Notably, immunofluorescence co-localization staining and pull-down assays confirmed that ELAVL1 directly interacts with APP. Collectively, these findings provide valuable insights into the molecular mechanisms underlying AD and suggest ELAVL1 as a promising therapeutic target for AD.
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TALANTA
·
2025
·
IF 6.1
Lingyan Kang,Tao Xu,Zhimeng Fan,Yiqi Li,Ye Tian
Achieving multiplexed, equipment-free, and sensitivity-enhanced antigen detection within a true one-step lateral flow format remains a technical challenge. Lateral flow immunoassays (LFIAs) are widely used for point-of-care diagnostics, yet their sensitivity—particularly in weak-positive specimens—often limits reliable pathogen identification. Herein, we enhance the analytical performance of LFIA by introducing a structurally integrated self-enhancement strategy that maintains the simplicity of one-step operation without requiring additional user steps. In the proposed SET-LFIA, two enhancement pads loaded with Au 3+ and NH 2 OH·HCl are positioned beneath the detection layer and isolated by a double-sided adhesive barrier, enabling sequential reagent release along the capillary flow path and inducing localized in situ gold growth during the post-binding phase. This design strengthens the visual signal while preserving the immunoreaction process. Under optimized conditions, the SET-LFIA achieved detection limits of 1.0 ng/mL for IAV, 1.0 ng/mL for IBV, and 0.2 ng/mL for SARS-CoV-2, corresponding to 5–25-fold sensitivity gains over conventional LFIA. Tests using spiked samples demonstrated markedly improved discrimination of weak-positive specimens and reduced false-negative outcomes. In addition, stability evaluation showed >85 % signal retention after 21 days at 45 °C, and clinical samples further confirmed robust multiplex detection across all three targets. With its enhanced sensitivity, one-step workflow, and compatibility with industrial lamination, the SET-LFIA represents a practical and scalable solution for rapid on-site respiratory pathogen screening.
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