ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Danping Chen,Yang Wang,Jun Chen,Yanfei Zhang,Zong Dai,Xiaoyong Zou
Active DNA demethylation process critically relies on the intrinsic properties of ten-eleven translocation proteins (Tets), particularly the flanking sequence preference. Challenged by the fact that the proximate bases to the 5-methylcytosine (5mC) are multitudinous and their influence on the Tets/DNA interplay is minute, the current methodologies are very limited in terms of cost, sensitivity and efficiency. Herein, we propose a polymerization retardation isothermal amplification (PRIA) strategy that enables sensitive and fast study of the flanking sequence preference of Tet. By arranging DNA polymerase to repetitively pass DNA strands through an isothermal replication-scission amplification reaction, the tiny difference in the Tet/DNA interplay can be consecutively accumulated and amplified. Low amount sample (80 ng) even multiple samples can be simultaneously analyzed within 10 h on an easily accessible laboratory real-time quantitative PCR instrument. For a proof-of-concept study, the binding preference ( P B ) of Tet2 for XmCGX, (X = C, G, T , A) was analyzed by PRIA and computational analysis, showing an order of AmCGT > TmCGA ≈ GmCGC > CmCGG. Furthermore, the binding and oxidation preference in Tet/DNA interplay process was individually considered. By comparative evaluation of the total flanking sequence preference ( P T ) and the P B , for the first time, we revealed that Tet2 has a lower oxidation preference ( P O ) to proximal flanking bases and the main contributor to P T of Tet2 is P B . The PRIA strategy, due to its reliable, cost-effective, high efficiency and low sample input, would hopefully advance epigenetic researches and other relative studies.
研究领域:
分子生物学
·
表观遗传学
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF FISH BIOLOGY
·
2016
·Vol. 88
·期 5
·1693-1707
·
IF 1.246
W. T. Zhang,K. Liu,J. S. Xiang,L. Y. Zhang,W. J. Liu,Z. D. Dong,Y. Z. Li,H. L. Li,S. L. Chen,N. Wang
To elucidate the effect of thyroid hormone receptor α A ( thraa ) on metamorphosis, the full length cDNA of half-smooth tongue sole Cynoglossus semilaevis was cloned. The relative gene transcript level of thraa at different development stages was quantified using real-time PCR. Transcription of thraa increased and declined rapidly during metamorphosis. Hyperthyroidism was induced in juveniles and larvae with exposure to T3 and T4, and hypothyroidism with thiourea (TU), 2-mercapto-1-methylimidazole (MMI). thraa mRNA was higher in fish treated for 6 days with MMI than in untreated controls, although inhibited larvae did not complete metamorphosis. The addition of exogenous T4 reversed this effect in the MMI-treated group, but not in the TU-treated group. In situ hybridization revealed progressive tail end of body growth and change during developmental stages, with corresponding changes in thraa expression. This process may be induced by thyroid hormones with thraa as a major mediator. The morphological changes of tip of the tail may be associated with the development of lateral swimming.
研究领域:
分子生物学
·
内分泌学
·
发育生物学
·
水产养殖
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Jiasheng Yuan,Jiayin Shen,Mingyu Chen,Zhichao Lou,Shuye Zhang,Zhigang Song,Weiwei Li,Xin Zhou
Direct visual enumeration of viruses under dark-field microscope (DFM) using plasmon resonance probes (PRPs) is fast and convenient; however, it is greatly limited in the assay of real samples because of its inability to accurately identify false positives owing to non-specific adsorption. In this study, we propose an artificial intelligence (AI)-assisted DFM enumeration strategy for the accurate assay of Enterovirus A71 (an ultra-small human virus) using two PRPs; a 40 nm silver nanoparticle probe (SNP) that appears bright blue under DFM, and a 120 nm gold nanorod probe (GNP) that appears red under DFM. The capture chip was prepared by immobilizing the SNPs with antibodies on the glass to capture the target virus and to form dichromatic sandwich structures with the GNPs, followed by imaging under a dark field (DF). Subsequently, the DF images of the capture chip were subjected to a two-step screening: first, using image processing , and thereafter using the AI algorithm screening to eliminate false positive results and background noise. The results revealed that the data from the AI-assisted dual PRPs assay were highly consistent with those of quantitative PCR (qPCR), and that the sensitivity with a minimum detectable concentration of 3 copies/μL was 5 times higher than that of qPCR. The entire analysis was completed within 45 min. Therefore, our AI-assisted virus enumeration strategy with two DF PRPs holds great potential for ultra-sensitive and accurate quantification of viruses in real samples.
研究领域:
病毒学
·
纳米技术
·
人工智能
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Evidence-based Complementary and Alternative Medicine
·
2022
·
IF 2.65
Qiang Shi,Jian-Gang Ju
Circular RNA (circRNA) is considered an important regulator of cancer. Circ_0001998 is a newly discovered circRNA and its role in lung adenocarcinoma (LUAD) remains obscure and requires further study. The expression levels of circ_0001998 and miR-145 in LUAD were predicted by bioinformatics analysis and then verified by qRT-PCR in the LUAD cell lines. CCK-8, clone formation, EdU assay, and flow cytometry were applied to determine the effects of silencing circ_0001998 on the viability, proliferation, and apoptosis of LUAD cells. The target relationship between circ_0001998 and miR-145 was predicted by bioinformatics analysis and verified by a luciferase activity experiment. The effect of circ_0001998/miR-145 axis on the viability, proliferation, and apoptosis of LUAD cells was verified by the rescue experiment. Circ_0001998 was upregulated in LUAD, and silencing circ_0001998 suppressed viability, proliferation, and invasion of LUAD cells. The target gene of circ_0001998, miR-145, was downregulated in LUAD, and the low expression of miR-145 indicated a poor prognosis. The effect of silencing circ_0001998 on the biological function of LUAD cells was reversed by the miR-145 inhibitor. Circ_0001998 regulates the proliferation, invasion, and apoptosis of LUAD via sponging miR-145.
研究领域:
肿瘤学
·
分子生物学
·
生物信息学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF THE INDIAN CHEMICAL SOCIETY
·
2022
·
IF 0.243
Xiao-Na Liang,Li-Ying Xu,Ming-Li Zhai,Jie Zhang
In the current research, through employing the dual ligand method, a novel coordination polymer has been produced in success via the reaction between H 2 glu and In(NO 3 ) 3 ·6H 2 O in the existence of the nitrogen-donor chelating 2,2′-bpy ligand in CH 3 CN In(III) and DMF mixed solvent, and its chemical formula is In(glu)(Hglu)(2,2′-bpy) ( 1 , 2,2′-bpy is 2,2′-bipyridine and H 2 glu is glutaric acid). Its application values on the acute stroke were assessed and the corresponding mechanism was investigated simultaneously. Firstly, the levels of inflammatory response in the astrocytes were evaluated by ELISA by measuring the content of inflammatory cytokines released into cerebrospinal fluid. Additionally, the miRNA199a relative expression levels in astrocytes were measured via exploiting real time RT-PCR. Molecular docking simulation demonstrated that synthesized In ion complex exhibited excellent biological activities, multiple binding interactions were formed by the carboxyl groups on the In ion complex.
研究领域:
药理学
·
神经科学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
FOOD CONTROL
·
2022
·
IF 6.652
Baoqing Zhou,Qinghua Ye,Moutong Chen,Chufang Wang,Xinran Xiang,Ying Li,Jumei Zhang,Youxiong Zhang,Juan Wang,Shi Wu,Qihui Gu,Yu Ding,Qingping Wu
Rapid and accurate screening of foodborne pathogens in contaminated food is with great significance for food safety. AuNPs-based colorimetric assay is an attractive strategy in the field of food safety inspection, but is limited by tedious probe preparation, low color rendering resolution, and narrow color gamut. Herein, we proposed an e nhanced c olorimetric p latform based on the C RISPR/Cas12a system and label-free D NA-AuNP p robe (CDP-ECP) for high-throughput detection of Staphylococcus aureus ST398. In this strategy, a novel and specific gene targeting S. aureus ST398 was identified and efficient AuNP-bioprobe was successfully prepared. Through a PCR/RAA amplification, the trans -ssDNA cleavage ability induced by target DNA product triggered an aggregation performance for the prepared AuNP-bioprobe pairs, which could realize target colorimetric detection within 50 min. Under optimal conditions, the colorimetric assay could detect as low as 6.4 × 10 2 cfu/mL of S. aureus ST398 in pure culture. Specific gene mining and target-induced trans -cleavage character gave this method a superior specificity and anti-interference. More importantly, this colorimetric method can be extended to a high-throughput platform for rapid detection of spiked and natural sample by using a common microplate reader. Since it is simple to operate, low cost, high sensitivity and specificity, this new platform is fully qualified for clinical diagnosis, environmental monitoring onsite testing requirements.
研究领域:
食品安全
·
分子生物学
·
生物技术
·
病原体检测
✓ 已验证
产品关联
DOI
高影响因子
Nanoscale
·
2023
·
IF 6.7
Haipeng Cui,Wenwei Pan,Tiechuan Li,Xiaotian Shen,Ye Chang,Wei Pang,Xuexin Duan
Virus infections remain one of the principal causes of morbidity and mortality worldwide. The current gold standard approach for diagnosing pathogens requires access to reverse transcription–polymerase chain reaction (RT‒PCR) technology. However, separation and enrichment of the targets from complex and diluted samples remains as a major challenge. In this work, we proposed a micromotor-based sample preparation concept for the efficient separation and concentration of target viral particles before PCR. The micromotors are functionalized with antibodies with a 3D polymer linker and are capable of self-propulsion by the catalytic generation of oxygen bubbles for selective and positive virus enrichment. This strategy significantly improves the enrichment efficiency and recovery rate of virus (up to 80% at 104 tu mL-1 in a 1 mL volume within just 6 min) without external mixing equipment. The method allows the Ct value in regular PCR tests appearing 6-7 cycles earlier and a detection limit of 1 tu mL-1 for target virus from swap samples. A point-of-need test kit is designed based on the micromotors which can be readily applied to pretreat a large volume of samples.
研究领域:
生物技术
·
病毒学
·
分析化学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
AMERICAN JOURNAL OF POTATO RESEARCH
·
2020
·Vol. 97
·期 4
·367-375
·
IF 0.858
Daurov Dias,Daurova Ainash,Karimov Aibek,Tolegenova Dana,Volkov Dmitriy,Raimbek Daniyar,Zhambakin Kabyl,Shamekova Malika
Potato ( Solanum tuberosum L.) is the fourth most common food crop worldwide and one of the main food products in Kazakhstan. One of the factors leading to crop loss in potato is viral disease. There are more than 30 viruses known to cause diseases in potato. Therefore, using virus-free planting material is an important aspect of growing potato. It is relatively difficult to eliminate viruses from plants, despite the application of isolated apical meristems. In this study, we used the most effective methods to eliminate viral infections, such as electrotherapy, chemotherapy (40 and 50 mg/l), cryotherapy, and thermotherapy. Complete elimination of certain viral infections, such as PVM, PVS, PVX, and PVY, was achieved after chemotherapy with 40 mg/l of ribavirin. A multiplex RT-PCR method was used for efficient monitoring of viral infections, which is more precise than the enzyme-linked immunosorbent assay (DAS-ELISA). The obtained virus-free planting material will subsequently be used for industrial virus-free potato seed production in Kazakhstan.
研究领域:
农业
·
植物病理学
·
病毒学
·
生物技术
✓ 已验证
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Yuanhuan Yang,Feng Gao,Ying Liang,Lichao Guo,Yang Pan,Peng Cao,Yue Zhang
The identification of Chinese medicinal herbs occupies a crucial part in the development of the food and drug market. Although molecular identification based on real-time PCR offers good versatility and uniform digital standards compared with traditional methods, such as morphology, the dependence on large-scale equipment hinders spot detection and marketable applications. In this study, we developed a DNA nanoclaw for colorimetric detection and visible on-site identification of Chinese medicines. When specific miRNA is present, the DNAzyme is activated and cleaves the substrate strand, triggering the catalytic hairpin assembly (CHA) reaction and forming branched DNA junctions on AuNP-I. This can then capture AuNP-II through hybridization and facilitate their aggregation, resulting in a noticeable color change that is observable to the naked eye. By harnessing the dual amplification of DNAzyme and CHA, this highly sensitive nanoprobe successfully achieved specific identification of Chinese medicines. This offers a new perspective for on-site testing in the herbal market.
研究领域:
生物技术
·
纳米技术
·
药学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Kehan Zhu,Zhen Yao,Tianyi Gu,Xianhao Jiang,Jianqin Zhou,Duxin Li
Swollen culms of Zizania latifolia possess a peculiar flavor, nutritional value and health-promoting effects. This study investigated the wound healing ability of polysaccharides isolated from Zizania latifolia (ZLP). Through in vitro assays, ZLP demonstrated negligible cytotoxicity and promoted the migration and phagocytosis of Staphylococcus aureus by macrophages where ZLP-2 had a significantly greater effect. Moreover, ZLP-2 has a higher purity and molecular weight, an integral and large lamellar morphology, and a larger particle size in solution, suggesting its ability to form films. Thus, ZLP-2 was fabricated as a gel formulation for topical application. The in vivo results showed that the ZLP-2 gel had significantly greater effects than did the autonomous healing process according to quantitative analysis of the wound closure area. The ZLP-2 gel decreased the levels of TNF-α and IL-6 in wounded tissue. Furthermore, the quantification of the molecular indexes via qRT-PCR revealed that the levels of tnf-α, tgf-β, mcp-1, ifn-γ, inos, and vegf in the treated group increased during the inflammatory and proliferation stages but decreased after wound closure. These results suggest that ZLP-2 has great potential in pharmaceutical applications for wound care.
研究领域:
药理学
·
伤口愈合
·
多糖研究
·
炎症
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LAB ON A CHIP
·
2021
·Vol. 21
·期 9
·1759-1770
·
IF 6.799
Wenwen Chen,Rongkai Cao,Wentao Su,Xu Zhang,Yuhai Xu,Peng Wang,Zhongqiao Gan,Yingying Xie,Hongjing Li,Jianhua Qin
Tumor-derived exosomes have been recognized as promising biomarkers for early-stage cancer diagnosis, tumor prognosis monitoring and individual medical treatment. However, it is a huge challenge to separate exosomes from trace biological samples in clinics for disease diagnosis. Herein, we propose a simple, quick, and label-free method for isolating circulating exosomes from serum of patients. The strategy synergistically integrates chitosan electrostatic-adsorption, micro-patterned substrates, and microfluidic shuttle flow control to enable the capture/release of circulating exosomes in a simple manner. Using this microchip, we can isolate exosomes from trace samples (10 μl) with relative purity over 90% and high RNA recovery ratio over 84% within 15 minutes, which is impossible for traditional ultracentrifugation methods. We then validate the application of the microchip using 24 serum samples from clinical breast cancer and breast fibroma patients. The isolated exosomes are subjected to miRNA sequencing and RT-PCR, followed by pathway prediction analysis. The results showed that exosomes were relevant to the invasion and metastasis of breast cancer cells and hsa-miR-18a-3p might have the potential to become a new biomarker for distinguishing breast cancer from breast fibroma (AUC = 0.83, P value = 0.019). This established method is simple, quick and easy to operate with integration. And it may pave a new way for clinical research on exosomes and tumor relevant diagnosis.
研究领域:
生物医学工程
·
癌症生物学
·
分子生物学
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Aquaculture Reports
·
2021
·
IF 3.216
Dongyong Fu,Yadong Hu,Peng Chu,Tao Wang,Mingxu Chu,Yonghai Shi,Shaowu Yin,Yongxiang Zhu,Yaohui Wang,Zhenglong Guo
Copper nanoparticles (Cu-NPs) are water pollutants, which has significant toxic environmental effects. However, the effects of Cu-NPs on Takifugu fasciatus are still relatively unknown. Compared with the control (no add CuSO 4 and Cu-NPs). This study aims to describe the pathological changes in the gill and liver of pufferfish exposed to CuSO 4 (Cu 2+ for short) and Cu-NPs dissolved at a concentration of either 20 or 100 μg/L, in a semi-static waterborne exposure regime for 28 days. All Cu treatments resulted in organ injuries, and the pathologies observed following exposure to Cu-NPs were broadly of the same type as those observed following exposure to Cu 2+ . In the gill, these included lamellar fusion, clubbed tips, and mucocytes swollen, and in the liver included oedema, aggregation of blood cells, and necrosis. Furthermore, enzyme activity assays revealed that Ca 2+ -ATP activity significantly increased under the influence of Cu, whilst Na + -K + -ATPase activity was significantly inhibited. In this study, the CRT in T. fasciatus (referred to as Tf CRT) was identified and full-length cDNA sequences were characterized. Subsequently, RT-PCR and western blot analysis were carried out to assess the expressions of Tf CRT in the gill and liver at both the mRNA and protein levels. Under both the Cu-NPs and Cu 2+ treatments, both the mRNA and protein expressions of Tf CRT were considerably higher in the gill and liver compared to that of the control treatment. Furthermore, there were no significant differences between Cu-NPs and Cu 2+ treatment (at the same treatment concentration) in the expressions of Tf CRT at the mRNA or protein levels. Overall, both Cu 2+ and Cu-NPs had negative effects on T. fasciatus , and Cu-NPs had the toxic effects similar to dissolved Cu. Our results provide the ecological safety data service of Cu-NPs in water, and provide a reference for the health T. fasciatus aquaculture.
研究领域:
环境毒理学
·
水生毒理学
·
分子生物学
·
鱼类生理学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）