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8 results on '"Chen, Ganghui"'

7. Label-Free Colorimetric Method for Detection of Vibrio parahaemolyticusby Trimming the G-Quadruplex DNAzyme with CRISPR/Cas12a

Author

Chen, Xueyun, Wang, Liu, He, Fang, Chen, Ganghui, Bai, Linlin, He, Kaiyu, Zhang, Fang, and Xu, Xiahong

Abstract

Vibrio parahaemolyticus(V. parahaemolyticus), which may cause gastrointestinal disorders in humans, is a pathogen commonly found in seafood. There are many methods for detecting V. parahaemolyticus, yet they have some shortcomings, such as high cost, labor-intensiveness, and complicated operation, which are impractical for resource-limited settings. Herein, we present a sequence-specific, label-free, and colorimetric method for visual detection of V. parahaemolyticus. This method utilizes CRISPR/Cas12a to specifically recognize the loop-mediated isothermal amplification (LAMP) products for further trans-cleaving the G-quadruplex DNAzyme and depriving its peroxidase-mimicking activity. In this way, the results can be directly observed with the naked eyes viathe color development of 2,2′-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS2–), which displays colorless for positive samples while green for target-free samples. We term such Cas12a–crRNA preventing ABTS2–from developing color by trimming the G-quadruplex DNAzyme as Cascade. The proposed method can detect 9.8 CFU (per reaction) of pure cultured V. parahaemolyticus, and the sensitivity is comparable to real-time LAMP. It has been applied for practical use and showed the capability to detect 6.1 × 102CFU/mL V. parahaemolyticusin shrimp samples. Based on this, the newly established Cascade method can be employed as a universal biosensing strategy for pathogenic bacterial testing in the field.

Published
2021
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8. Metal-DNA Nanocomplexes Enhance Chemo-dynamic Therapy by Inhibiting Autophagy-Mediated Resistance.

Author

Gu C, Liu X, Luo L, Chen J, Zhou X, Chen G, Huang X, Yu L, Chen Q, Yang Y, and Yang Y

Subjects
Animals, Mice, Cell Line, Tumor, Metals, Hydroxyl Radical metabolism, Autophagy, Hydrogen Peroxide metabolism, Tumor Microenvironment, DNA, Catalytic, Neoplasms pathology, Nanoparticles
Abstract

Chemo-dynamic therapy (CDT) based on the Fenton or Fenton-like reaction has emerged as a promising approach for cancer treatment. However, autophagy-mediated self-protection mechanisms of cancer cells pose a significant challenge to the efficacy of CDT. Herein, we developed metal-DNA nanocomplexes (DACs-Mn) to enhance CDT via DNAzyme inhibition of autophagy. Specifically, Mn-based catalyst in DACs-Mn was used to generate highly hydroxyl radicals (⋅OH) that kill cancer cells, while the ATG5 DNAzyme incorporated into DACs-Mn inhibited the expression of autophagy-associated proteins, thereby improving the efficacy of CDT. By disrupting the self-protective pathway of cells under severe oxidative stress, this novel approach of DACs-Mn was found to synergistically enhance CDT in both in vitro and in vivo models, effectively amplifying tumor-specific oxidative damage. Notably, the Metal-DNA nanocomplexes can also induce immunogenic cell death (ICD), thereby inhibiting tumor metastasis. Specifically, in a bilateral tumor model in mice, the combined approach of CDT and autophagy inhibition followed by immune checkpoint blockade therapy shown significant potential as a novel and effective treatment modality for primary and metastatic tumors., (© 2023 Wiley-VCH GmbH.)

Published
2023
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