DYES AND PIGMENTS
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2019
·
IF 4.018
Muhammad Sohail Haroone,Yuehua Hu,Ping Zhang,Ruili Ma,Xiaorang Zhang,Qari Muhammad Kaleem,Muhammad Bashir Khan,Jun Lu
Nucleic acids , the hereditary material of living beings, perform a vital role in major life activities including growth, heredity, and variation. The genetic information is coded in specific nitrogenous bases , which are the key players in genetic transformation and identification of hereditary diseases. Telomeres are specialized DNA structures present at the end of eukaryotic chromosomes, consisting of repetitive non-transcribed sequences (TTAGGG) and some binding proteins . They protect the chromosomal ends from sticking to each other, and play a key role in the maintenance of genomic integrity. Currently, a number of methods are reported for nucleic acids and telomeres detection but they are not easy going. Therefore, development of sensitive and fast DNA-biosensors to measure the nucleic acids and telomeres sequences is of great importance. This paper describes a systematic study on the preparation of simple ultrathin films (UTFs) composite of fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI), having binding properties in the minor groove of AT-rich DNA segments, blending with single-stranded DNA (ssDNA) and incorporation into layered double hydroxides (LDH) nanosheets using layer-by-layer self-assembly method. The resulting UTFs exhibits excellent sensitivity, selectivity and reversibility for long complementary ssDNA sequence on the basis of DNA hybridization and fluorescence enhancement strategy. The dynamic range was 3–20 μg/mL with a detection limit of 20 μg/mL. This new (ssDNA @DAPI/LDHs) n UTFs, could be an efficient hybrid composite with potential applications in the field of bioluminescent sensoring materials.
研究领域:
生物化学
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分子生物学
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Yu Ling,Jiao Zhou,Xiao Fang Zhang,Xiao Hu Wang,Nian Bing Li,Hong Qun Luo
As one of critical base excision repair enzymes, uracil-DNA glycosylase (UDG) can specifically repair uracil-induced DNA damage to maintain the genome integrity of organisms. Moreover, abnormal expression of UDG is related to various cancers and other serious diseases. Therefore, it is essential to accurately and sensitively monitor UDG activity. In this work, a ratiometric fluorescence method using poly-(thymine) DNA template copper nanoclusters (Cu NCs) and 4′,6-diamidino-2-phenylindole (DAPI) as the output signals is developed for simple, selective, and sensitive detection of UDG. A double-stranded DNA (dsDNA) consisting of a uracil-containing single chain and a trigger sequence is designed as a substrate dsDNA. Meanwhile, another dsDNA comprising 30 pairs of A-T bases ((AT) 30 dsDNA) is designed to be partially complementary to the trigger chain. The (AT) 30 dsDNA itself has no blunt or recessed 3′ end and cannot be cut by exonuclease III (Exo III). With the Exo III-assisted amplification, adding UDG results in the success synthesis of red-emissive Cu NCs, while the dsDNAs in the system are hydrolyzed and the fluorescence signal of DAPI is weak. In contrast, without addition of UDG, the fluorescence intensity of Cu NCs is low, while the DAPI emits significantly enhanced fluorescence signal. The proposed method presents a detection limit of 5.0 × 10 −5 U mL −1 and can be applied to the HeLa cell lysate sample with satisfactory results. Additionally, the kinetic parameter is studied and the UDG inhibition effect is evaluated. In summary, this work provides a potential platform for DNA repair enzyme-related analysis and clinical diagnosis.
研究领域:
生物化学
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分子生物学
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癌症研究
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分析化学
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高影响因子
Micromachines
·
2017
·Vol. 8
·期 2
·36
·
IF 1.833
Jing Sun,Jiayu Zhang,Haibo Yang,Gongzhuo Wang,Yanzhao Li,Xuxin Zhang,Qidan Chen,Ming-Fei Lang
Single-cell cell cycle analysis is an emerging technique that requires detailed exploration of the image analysis process. In this study, we established a microfluidic single-cell cell cycle analysis method that can analyze cells in small numbers and in situ on a microfluidic chip. In addition, factors that influenced the analysis were carefully investigated. U87 or HeLa cells were seeded and attached to microfluidic channels before measurement. Cell nucleic DNA was imaged by 4′-6-diamidino-2-phenylindole (DAPI) staining under a fluorescent microscope and subsequently fluorescent intensities of the cell nuclei DNA were converted to depict histograms for cell cycle phases. DAPI concentration, microscopic magnification, exposure time and cell number were examined for optimal cell cycle analysis conditions. The results showed that as few as a few hundred cells could be measured by DAPI staining in the range of 0.4–0.6 μg/mL to depict histograms with typical cell cycle phase distribution. Microscopic magnification during image acquisition, however, could distort the phase distribution. Exposure time did not significantly affect the cell cycle analysis. Furthermore, cell cycle inhibitor rapamycin treatment changed the cell cycle phase distribution as expected. In conclusion, a method for microfluidic single-cell cell cycle analysis of spread cells in situ was developed. Factors such as dye concentration and microscopic magnification had more influence on cell cycle phase distribution. Further studies will focus on detail differentiation of cell cycle phases and the application of such a method for biological meanings.Keywords:microfluidics;cell cycle;spread cell;DNA content
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高影响因子
ANALYTICAL LETTERS
·
2024
·
IF 1.6
Tianhan Zhang,Chengyi Xie,Yong Wu,Wenqing Gao,Chenlu Wang,Keqi Tang,Jiancheng Yu
A compact in-trap single photon ionization (SPI) system, termed discontinuous atmospheric pressure interface (DAPI) ion trap mass spectrometry (ITMS), for the rapid determination of trace volatile organic compounds (VOCs). DAPI enables the maintenance of high analyte pressure in the SPI source while preserving the vacuum. Utilizing vacuum ultraviolet (VUV) lamp as the ion source, we ionize the analyte within an enclosed miniature 3D ion trap for efficient analysis. Experimental findings demonstrate that the system achieves a mass resolution of less than 0.5 atomic mass units for full width at half maxima (FWHM), providing unit resolution. For the response for benzene, the system achieves almost instantaneous response, with rise and fall times of only 2 s, which is a significant advantage compared to the membrane inlet (MI) approach necessary for on-site detection. Additionally, tandem mass spectrometry (MS/MS) with unit m/z isolation window was achieved and resolved dimethyl disulfide and phenol with similar molecular weights. Our results demonstrate that the system provides excellent performance and we anticipate its application in rapidly determining VOCs.
研究领域:
分析化学
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环境科学
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DOI
高影响因子
Cancer Cell International
·
2021
·Vol. 21
·期 1
·1-12
·
IF 5.722
Jiang Dongxiao,Ding Shufei,Mao Zhujun,You Liyan,Ruan Yeping
Background Colon cancer is a malignant gastrointestinal tumour with high incidence, mortality and metastasis rates worldwide. Aloe-emodin is a monomer compound derived from hydroxyanthraquinone. Aloe-emodin produces a wide range of antitumour effects and is produced by rhubarb, aloe and other herbs. However, the mechanism by which aloe-emodin influences colon cancer is still unclear. We hope these findings will lead to the development of a new therapeutic strategy for the treatment of colon cancer in the clinic.Methods We identified the overlapping targets of aloe-emodin and colon cancer and performed protein–protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. In addition, we selected apoptosis pathways for experimental verification with cell viability, cell proliferation, caspase-3 activity, DAPI staining, cell cycle and western blotting analyses to evaluate the apoptotic effect of aloe-emodin on colon cancer cells.Results The MTT assay and cell colony formation assay showed that aloe-emodin inhibited cell proliferation. DAPI staining confirmed that aloe-emodin induced apoptosis. Aloe-emodin upregulated the protein level of Bax and decreased the expression of Bcl-2, which activates caspase-3 and caspase-9. Furthermore, the protein expression level of cytochrome C increased in a time-dependent manner in the cytoplasm but decreased in a time-dependent manner in the mitochondria.Conclusion These results indicate that aloe-emodin may induce the apoptosis of human colon cancer cells through mitochondria-related pathways.
研究领域:
肿瘤学
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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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DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
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2020
·
IF 2.877
Ao Li,Peng Zuo,Bang-Ce Ye
Salmonella, a typical foodborne pathogen, always seriously threatens the health and even life of both humans and animals. However, highly sensitive and fast quantitative methods for its detection are remaining to be challenged. Herein, we presented an efficient method with dual signal amplification strategy by combining immune hybridization chain reaction (HCR) with surface enhanced Raman scattering (SERS) to high sensitively detect Salmonella typhimurium in food. After sample preparation, S. typhimurium were specifically captured by immunomagnetic beads (IMBs), then aptamers and hairpin-probes were added to trigger HCR to form nicked dsDNA, finally 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) was incubated with HCR products and then the whole system was mixed with AgNP colloid to detect the SERS intensity at 1610 cm −1 . As a result, a good linear relationship was achieved between SERS intensities and corresponding concentrations of S. typhimurium ranging from 10 to 10 5 CFU/mL, with a limit of detection (LOD) of 6 CFU/mL in 3.5 h. The proposed method has been successfully applied to capture and detect the S. typhimurium in spiked milk samples, and the results were consistent with those of the traditional plate counting method. The method, with combination of HCR and SERS, achieves double amplification of the detection signal and significantly improves the detection sensitivity of S. typhimurium . And it also shows good application potential for the highly sensitive detection of other contaminants in food.
研究领域:
食品科学
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生物传感器
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病原体检测
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分析化学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY
·
2017
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IF 3.505
Jun Tian,Zhaoqun Lu,Yanzhen Wang,Man Zhang,Xueyan Wang,Xudong Tang,Xue Peng,Hong Zeng
The antifungal activity of Nerol (NEL) against Candida albicans , a pathogenic fungus, has a minimum inhibitory concentration (MIC) of 4.4 mM that causes noteworthy candidacidal activity through an apoptosis-like mechanism. Calcium (Ca 2+ ) levels and reactive oxygen species (ROS) production, which are the major causes of apoptosis, were determined in C. albicans cells treated with NEL and were found to increase, which related to mitochondrial dysfunction and disruption. A series of characteristic changes of apoptosis caused by NEL, including mitochondrial membrane depolarization, cytochrome c (cyt c) release, and metacaspase activation were examined using a flow cytometer and Western blot . The results showed that an increase in intracellular Ca 2+ and ROS led to dramatically decreased mitochondrial membrane potential (MMP); cyt c was also released from the mitochondria to the cytosol. Other early apoptotic features were also observed with the metacaspase activation. Finally, the morphological changes of the cells were observed, including phosphatidylserine (PS) externalization, nuclear condensation, and DNA fragmentation through Annexin V-FITC and PI double staining, TUNEL assay , and DAPI staining. The results supported the hypothesis that NEL was involved in the apoptosis of C. albicans cells not only at the early stages, but also at the late stages. In summary, NEL can trigger mitochondrial dysfunction and disruption via elevation of Ca 2+ and ROS leading to apoptosis in C. albicans . This research on the mechanism of cell death triggered by NEL against C. albicans has important significance for providing a novel treatment of C. albicans infections.
研究领域:
微生物学
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药理学
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细胞生物学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Jingbo Sun,Hongchen Li,Min Ouyang,Jing Cheng,Dong Xu,Xiaofeng Tan,Qinlu Lin
Cytoarchitectural staining is of great importance in disease diagnosis and cell biology research. This study developed user-friendly multifunctional red-emissive carbon dots (R-CDs) for rapid cell nucleus staining via targeting nuclear proteins. R-CDs, simply prepared by electrochemical treatment of 1,2,4-benzenetriamine, exhibit strong emission at 635 nm when excited at 507 nm. The R-CDs can rapidly stain the nucleus of human SH-SY5Y, HepG2, and HUH-7 cells with a high signal-to-noise ratio owing to fluorescence enhancement after entering the nucleus. Compared to conventional cytosolic dyes such as Hoechst and DAPI, R-CDs are cheaper, more highly dispersed in water, and more stable (requiring no stringent storage conditions). The R-CDs show stable optical properties with insignificant photobleaching over 7 days and salt resistance up to 2 M of NaCl. More importantly, R-CDs, possessing a positive charge, allow rapid staining of live cells (3 min) and dead cells (10 s) in saline. According to kinetic variation, R-CDs can distinguish live cells from dead cells. Staining exhibits high efficiency in onion epidermal cells, Aspergillus niger, Caenorhabditis elegans, and human spermatozoa. The mechanism for efficient staining is based on their fast accumulation in the nucleus due to their small size and positive charge and strong interaction with nuclear proteins at amino acid residues of histidine and arginine, resulting in fluorescence enhancement by dozens of times. The developed R-CDs do not bind to DNA and would not cause genetic damage and will find various safe applications in biological and medical fields.
研究领域:
细胞生物学
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疾病诊断
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材料科学
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光学性质
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生物医学应用
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DOI
PubMed
高影响因子
BMC Cardiovascular Disorders
·
2024
·Vol. 24
·期 1
·1-13
·
IF 2
Wang Xin,Sun Limin
MI (myocardial infarction) often triggers severe heart failure and is one of the leading causes of death worldwide. Receptor expression-enhancing protein 5 (REEP5), a member of REEPs, acts as regulators of endoplasmic reticulum (ER) affecting cardiac functions. Based on GSE114695 profile data, REEP5 was decreased in the left ventricle of MI mice. However, its role and potential mechanism in MI remain to be investigated. In the present study, the mouse MI model was established by ligation of the left anterior descending artery. REEP5 expression was downregulated in the infarct penumbra area of MI mice. Next, its role during MI was explored by gain-of-function. Interestingly, REEP5 overexpression improved left ventricular function of mice with MI, accompanied with reduced infarct size. In cardiomyocytes, REEP5 overexpression inhibited ER stress, accompanied with repressive phosphorylation of PERK and IRE1α, and the decreased nuclear translocation of ATF6. Subsequently, REEP5 overexpression downregulated the levels of Chop and cleaved caspase-12, further alleviating ER stress-induced apoptosis, which was consistent with the in vivo results. Moreover, REEP5 was found to bind to C-type lectin member 5 A (CLEC5A), a protein that triggers cardiac dysfunction. CLEC5A, whose expression was elevated in hypoxia-induced cell models, led to cardiomyocyte apoptosis. Noteworthily, REEP5 overexpression markedly abolished the effects of CLEC5A on ER stress-induced apoptosis. Taken together, REEP5 mediated the function of CLEC5A to relieve MI via inhibiting ER stress-induced apoptosis in vivo and in vitro. REEP5 may be a promising target for treating MI.
研究领域:
心脏病学
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分子生物学
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细胞生物学
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病理学
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DOI
PubMed
高影响因子
Heliyon
·
2024
·
IF 3.4
Chongchong Liu,Jiangyan Sun,Xiaoming Shen,Shefang Li,Sha Luo,Na Chen,Yunke Zhang
Epilepsy is a chronic neurological disorder. Drug-resistant epilepsy (DRE) accounts for about one-third of epilepsy patients worldwide. Peimine, a main active component of Fritillaria, has been reported to show anti-inflammatory effects. However, its potential therapeutic role in DRE is not yet fully understood. In this work, a DRE rat model was established by injecting 1 μg kainic acid (KA), followed by a 250 mg/kg administration of valproic acid (VPA) from day 4–31. Rats were treated with different doses of peimine (2.5 mg/kg, 5 mg/kg and 10 mg/kg) daily from day 32–62. In vitro, BV-2 microglia were exposed to different doses of peimine (7.5 μg/ml, 15 μg/ml, and 30 μg/ml) in presence of LPS. The aim of this study was to investigate the potential therapeutic effects of peimine on DRE. The results showed that peimine efficiently suppressed the KA-induced epileptic behaviors of rats in a dose-dependent manner, as recorded by electroencephalography. Furthermore, peimine ameliorated hippocampal neuron injury in DRE rats, and promoted an M1-to-M2 microglial phenotype shift in a dose-dependent manner. Mechanistically, peimine inhibited the TLR4/NF-κB/HIF-1α signaling pathway both in vivo and in vitro. Additionally, peimine suppressed the apoptosis of primary neurons induced by LPS-treated microglia. In conclusion, peimine augments the microglial polarization towards an M2 phenotype by inhibiting the TLR4/NF-κB/HIF-1α signaling pathway, thereby attenuating DRE.
研究领域:
神经病学
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药理学
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免疫学
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细胞生物学
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DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR HISTOLOGY
·
2025
·Vol. 56
·期 2
·1-11
·
IF 2.2
Zhang Xiuping,Wang Shimin,Liang Lixia,Hu Fen,Zhang Xueluo,Cui Xiangrong,Zhang Zhiping,Wu Xueqing
Recurrent spontaneous abortion (RSA) brings tremendous difficulties to clinical treatment and prognosis. Pre-B-cell leukemia homeobox 1 (PBX1), as a functional transcription factor, involves in the regulation of cell apoptosis and proliferation. However, the underlying mechanism of PBX1 in RSA treatment has not been explored. We established a lipopolysaccharide (LPS)-induced abortion model and detected PBX1 expression with real-time PCR, western blot and immunohistochemistry. The PBX1-overexpressed adenovirus (AV-oePBX1) was infected into trophoblast cells treated with LPS to define the function of PBX1 on cell apoptosis, inflammation and NF-κB pathway. A luciferase reporter assay was conducted to validate the transcription regulation of PBX1 on transmembrane and ubiquitin like domain containing 1 (TMUB1). Compared to women with normal abortion, PBX1 was downregulated in the placental villous tissues of RSA patients. The placental tissues of LPS-treated mice also manifested notably reduction of PBX1 at mRNA and protein levels. PBX1 overexpression alleviated inflammation and apoptosis of trophoblast cells. Substantially, PBX1 was negatively correlated with TMUB1, which was highly expressed in RSA patients and LPS-treated mice. Moreover, PBX1 bound to TMUB1 promoter and inhibited its transcription. Interestingly, exogenous TMUB1 abolished the effects of PBX1 on apoptosis, inflammation, and NF-κB signal pathway. In total, PBX1 attenuated cell apoptosis and inflammation, and suppressed NF‐κB signal pathway induced by LPS through downregulating TMUB1 transcription. Therefore, PBX1 may be developed as a novel target for clinical treatment of RSA.
研究领域:
妇产科学
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分子生物学
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转化医学
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DOI
PubMed
高影响因子
Nanoscale
·
2024
·Vol. 16
·期 8
·4095-4104
·
IF 5.8
Xinyi Cai,Tiantian Xu,Rui Ding,Dou Zhang,Guiquan Chen,Wenchang Zhao,Jiajie Hou,Hong Pan,Qian Zhang,Ting Yin
In recent years, tumor catalytic therapy based on nanozymes has attracted widespread attention. However, its application is limited by the tumor hypoxic microenvironment (TME). In this study, we developed oxygen-supplying magnetic bead nanozymes that integrate hemoglobin and encapsulate the photosensitizer curcumin, demonstrating reactive oxygen species (ROS)-induced synergistic breast cancer therapy. Fe3O4 magnetic bead-mediated catalytic dynamic therapy (CDT) generates hydroxyl radicals (˙OH) through the Fenton reaction in the tumor microenvironment. The Hb-encapsulated Fe3O4 magnetic beads can be co-loaded with the photosensitizer/chemotherapeutic agent curcumin (cur), resulting in Fe3O4-Hb@cur. Under hypoxic conditions, oxygen molecules are released from Fe3O4-Hb@cur to overcome the TME hypoxia, resulting in comprehensive effects favoring anti-tumor responses. Upon near-infrared (NIR) irradiation, Fe3O4-Hb@cur activates the surrounding molecular oxygen to generate a certain amount of singlet oxygen (1O2), which is utilized for photodynamic therapy (PDT) in cancer treatment. Meanwhile, we validated that the O2 carried by Hb significantly enhances the intracellular ROS level, intensifying the catalytic therapy mediated by Fe3O4 magnetic beads and inflicting lethal damage to cancer cells, effectively inhibiting tumor growth. Therefore, significant in vivo synergistic therapeutic effects can be achieved through catalytic–photodynamic combination therapy.
研究领域:
生物医学工程
·
肿瘤学
·
纳米医学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）