NEW JOURNAL OF CHEMISTRY
·
2023
·Vol. 47
·期 28
·13228-13234
·
IF 3.3
Yuanyi Wu,Shihan Liu,Yumei Xiao,Qian Li,Qin Xiang,Xianqin Song,Shanglin Wu,Kaiyong Wang,Yi Yang
Accurate detection of DNA is critical for biomedicine and food safety. Layered double hydroxides (LDHs) are a class of two-dimensional layered materials that have received extensive attention in the fields of sensors, catalysis, and materials science. The application of LDHs in DNA sensing in turn represents a fascinating topic. In this study, we found that both 6-carboxyfluorescein (FAM)-modified single-stranded DNA (ssDNA) probes and probe/target double-stranded DNA (dsDNA) duplexes could be adsorbed by LDHs, which resulted in high fluorescence quenching (>95%) for the first time. Based on the extraordinarily high quenching efficiency of LDHs, we developed a fluorescent sensing platform for simple and highly efficient detection of DNA, using Ni/Al-LDHs as fluorescence quenchers. For the selective degradation of dsDNA, exonuclease III was integrated into the sensor. In the sensing platform, the fluorescence of the probes was rapidly quenched by Ni/Al-LDHs. In the presence of the target DNA, dsDNA was formed, which was subsequently selectively degraded by exonuclease III into short fragments, resulting in the maintenance of the fluorescence of FAM labels. The linear range was 10–400 nM. The detection limit was 1.38 nM. This sensor accomplished the detection of target DNA within 50 min. It provides a convenient, rapid and versatile sensing platform for DNA biomarkers.
研究领域:
生物医学工程
·
材料科学
·
分析化学
·
生物传感器
✓ 已验证
DOI
高影响因子
Advanced Science
·
2025
·
IF 14.1
Wei Duan,Rendy Hosea,Lingxian Wang,Cao Ruan,Fuqiang Zhao,Jingyi Liu,Hezhao Zhao,Makoto Miyagishi,Shourong Wu,Vivi Kasim
Immune checkpoint inhibitor (ICI) therapy is a promising anti-tumor therapeutic strategy; however, its efficacy in solid tumors is limited. Chromosome missegregation is common in various solid tumors; however, its role in tumor progression remains poorly understood, and its correlation with ICI is yet to be explored. Here, it is found that increased chromosome missegregation promotes tumor immune microenvironment, and eventually immunotherapeutic efficacy, by triggering pyroptosis. yin yang 2 (YY2) is identified as a mitotic checkpoint regulator, which promotes chromosome missegregation by upregulating BUB1B transcription. Increased chromosome missegregation promoted the formation of micronuclei and release of double-stranded DNA (dsDNA) into the cytosol, triggering an AIM2-mediated cytosolic dsDNA response. The subsequent pyroptosis strengthened the tumor immune microenvironment, thereby enhancing immunoinfiltration and cytotoxicity of CD8 + T cells, while preventing their exhaustion. Finally, through in vitro and in vivo experiments, it is demonstrated that combining YY2 overexpression-induced chromosome missegregation/cytosolic dsDNA response and PD-1 inhibitor significantly enhanced the efficacy of ICI immunotherapy in microsatellite instable and microsatellite stable colorectal cancer cells. Together, these findings provide new insights on the role of chromosome missegregation in triggering cytosolic dsDNA response-mediated pyroptosis and modulating the tumor immune microenvironment, suggesting a novel strategy for improving ICI therapeutic efficacy in colorectal cancer.
研究领域:
肿瘤学
·
免疫学
·
分子生物学
·
癌症治疗
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Viruses-Basel
·
2024
·Vol. 16
·期 5
·768
·
IF 3.8
Sixu Liu,Jingqi Li,Qingtian Cheng,Kangyi Duan,Zhan Wang,Shuang Yan,Shuaishuai Tian,Hairui Wang,Shaobin Wu,Xinkui Lei,Yu Yang,Ningning Ma
Influenza vaccines, which are recommended by the World Health Organization (WHO), are the most effective preventive measure against influenza virus infection. Madin–Darby canine kidney (MDCK) cell culture is an emerging technology used to produce influenza vaccines. One challenge when purifying influenza vaccines using this cell culture system is to efficiently remove impurities, especially host cell double-stranded DNA (dsDNA) and host cell proteins (HCPs), for safety assurance. In this study, we optimized ion-exchange chromatography methods to harvest influenza viruses from an MDCK cell culture broth, the first step in influenza vaccine purification. Bind/elute was chosen as the mode of operation for simplicity. The anion-exchange Q chromatography method was able to efficiently remove dsDNA and HCPs, but the recovery rate for influenza viruses was low. However, the cation-exchange SP process was able to simultaneously achieve high dsDNA and HCP removal and high influenza virus recovery. For the SP process to work, the clarified cell culture broth needed to be diluted to reduce its ionic strength, and the optimal dilution rate was determined to be 1:2 with purified water. The SP process yielded a virus recovery rate exceeding 90%, as measured using a hemagglutination units (HAUs) assay, with removal efficiencies over 97% for HCPs and over 99% for dsDNA. Furthermore, the general applicability of the SP chromatography method was demonstrated with seven strains of influenza viruses recommended for seasonal influenza vaccine production, including H1N1, H3N2, B (Victoria), and B (Yamagata) strains, indicating that the SP process could be utilized as a platform process. The SP process developed in this study showed four advantages: (1) simple operation, (2) a high recovery rate for influenza viruses, (3) a high removal rate for major impurities, and (4) general applicability.
研究领域:
生物技术
·
病毒学
·
药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CONTROL
·
2022
·
IF 6.652
Yujiao Hou,Nan Long,Boyu Jia,Xiaofang Liao,Minghan Yang,Lizhu Fu,Lidong Zhou,Ping Sheng,Weijun Kong
The prevalent ochratoxin A (OTA) contamination in food has increased the urgency to develop sensitive and accurate detection methods. This study reported a conveniently fabricated label-free electrochemical (EC) aptasensor for ultrasensitive detection of OTA based on the competitive binding of OTA with its specific aptamer without any signal amplification strategy. A gold electrode (AuE) was modified with a compact layer of double-stranded DNA (dsDNA), obtained via OTA aptamer and complementary DNA (cDNA) hybridization, to act as a barrier for redox probe [Fe(CN) 6 ] 3-/4- to AuE access. This led to a high initial electron-transfer resistance (R ct ), recorded via electrochemical impedance spectroscopy (EIS). Since the aptamer displayed a higher affinity to OTA than cDNA, the added OTA triggered aptamer dissociation from AuE to form the OTA-aptamer complex, obviously decreasing the R ct . The target-induced variation in the resistance response strongly depended on the OTA concentration. Optimum conditions improved the linear relationship between R ct and the logarithm of OTA concentration in a range of 0.05–10 ng/mL with a limit of detection (LOD) of 0.05 ng/mL for OTA. Recoveries of 74.8–105% with relative standard deviations (RSDs) of 8.86–13.09% were obtained in the spiked malt samples. The Real-world detection of OTA in the actual malt samples confirmed the reliability of the convenient, cost-effective, and highly specific impedimetric aptasensor, exhibiting significant promise for extended application in food control to identify more hazardous contaminants by altering the corresponding aptamers.
研究领域:
食品科学
·
分析化学
·
电化学
·
毒理学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）