BIOMACROMOLECULES
·
2019
·
IF 5.667
Bingchao Duan,Siwei Zou,Ying Sun,Xiaojuan Xu
A nanoplatform carrying doxorubicin (Dox) for cancer therapy and a dye for imaging was developed based on a natural triple helix β-glucan (t-LNT) and polydeoxyadenylic acid (poly(dA)). The t-LNT-Dox conjugates were prepared through Schiff-base reaction between the aldehyde group in the oxidized t-LNT and the amino group of Dox, the single chains (s-LNT-Dox) of which interacted with the poly(dA)-dye to form a composite s-LNT-Dox/poly(dA)-dye through hydrogen bonding between s-LNT and poly(dA). t-LNT-Dox was confirmed to acid-responsively release Dox in vitro, showing enhanced cytotoxicity against HeLa cancer cells with time. It was confirmed that Dox and the dye could be simultaneously delivered into HeLa cells or the tumors with a prolonged duration time. Furthermore, LNT-Dox conjugates effectively inhibited tumor growth and decreased adverse effects of the free Dox in vivo. Hence, this work develops a new strategy to fabricate the nanoplatform for therapy and imaging using a natural polysaccharide.
研究领域:
生物材料学
·
肿瘤学
·
药剂学
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DOI
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Chumeng Wang,Xinyue Yin,Lu Zhang,Nengsheng Ye,Yuhong Xiang
Tobramycin (TOB) is a widely used aminoglycoside antibiotic for treating human and animal diseases. However, its overuse poses a threat to human health, necessitating the development of a rapid and simple detection method. In this study, polyadenine-aptamer-stabilized gold nanoclusters (A x -Apt-AuNCs) were synthesized, to investigate the impact of different polyadenine lengths on their properties. A 20 -Apt-AuNCs demonstrated efficient catalytic activity in the oxidation of o -phenylenediamine to 2,3-diaminophenazine (DAP) in the presence of hydrogen peroxide, resulting in yellow fluorescence emission. Upon binding specifically to the TOB aptamer on A 20 -Apt-AuNCs, TOB enhanced both their peroxidase-like activity and the fluorescence intensity of DAP. Based on this mechanism, a fluorescence-enhanced aptasensor was developed for TOB detection. The aptasensor exhibited a linear detection range of 10.0 nM to 1.0 μM, with a detection limit of 2.63 nM. Furthermore, its application in real sample analysis produced satisfactory results.
研究领域:
分析化学
·
生物医学工程
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
Nature Protocols
·
2022
·Vol. 17
·期 9
·1980-2007
·
IF 17.021
Liu Yusheng,Zhang Yiwei,Wang Jiaqiang,Lu Falong
Poly(A) tails are added to the 3′ ends of most mRNAs in a non-templated manner and play essential roles in post-transcriptional regulation, including mRNA export, stability and translation. Measuring poly(A) tails is critical for understanding their regulatory roles in almost every aspect of biological and medical studies. Previous methods for analyzing poly(A) tails require large amounts of input RNA (microgram-level total RNA), which limits their application. We recently developed a poly(A) inclusive full-length RNA isoform-sequencing method (PAIso-seq) at single-oocyte-level sensitivity (a single mammalian oocyte contains ~0.5 ng of total RNA) based on PacBio sequencing that enabled accurate measurement of the poly(A) tail length and non-A residues within the body of poly(A) tails along with the full-length cDNA, providing the opportunity to study precious in vivo samples with very limited input material. Here, we describe a detailed protocol for PAIso-seq library preparation from single mouse oocytes or bulk oocyte samples. In addition, we provide a complete bioinformatic pipeline to perform the analysis from the raw data to downstream analysis. The minimum time required is ~14.5 h for PAIso-seq double-stranded cDNA preparation, 2 d for PacBio sequencing in HiFi mode and 8 h for the initial data analysis.
研究领域:
分子生物学
·
基因组学
·
转录组学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Jiaona Xu,Chunying Wei
Two general and reliable fluorescence sensors were proposed in this work utilizing aptamer DNA-templated silver nanoclusters (Ag NCs). Both DNA-AgNCs could be used for label-free detecting of ATP with the limits of detection of 0.44 and 0.65 mM. One of them was further applied to monitor the activity of adenosine deaminase (ADA). In our effort to elucidate the light-up mechanism, we studied a total of six Ag NCs prepared by different DNA sequences , and found that they showed different sensitivity to ATP. Both BT3T3- and BT3T3(R)-templated Ag NCs were chose to make particular studies by UV–vis, TEM, fluorescence, and TCSPC methods. The results showed that when DNA-Ag NCs was kept for 1.5 h and presented a strong fluorescence, the addition of ATP failed to cause a large change of fluorescence intensity ; on the contrary, after Ag NCs was kept for 24 h and emitted a weak fluorescence, adding ATP was able to result in the large fluorescence enhanced of 43 and 33 times for BT3T3- and BT3T3(R)-templated Ag NCs, respectively. The possible mechanism was also suggested that ATP binding to aptamer segment of template induced the change of the DNA secondary structure , which made the aggregated Ag nanoparticles disperse into Ag NCs with an average diameter of about 2 nm that were responsible for the large fluorescence increase. Moreover, ATP could protect the fluorescence intensity of BT3T3(R)-templated Ag NCs from quenching for at least 9 h.
研究领域:
生物化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2022
·
IF 8.025
Feifei Wang,Gang Yu,Qing Yang,Xiao Yi,Linglin Fu,Yanbo Wang
A homogeneous polysaccharide (GAP), with a molecular weight of 51.8 kDa, was isolated from edible red seaweed Gelidium amansii . Composition analysis suggested GAP contained 5.31% sulfate and 17.33% 3,6-anhydro-galactose and was mainly composed of galactose . Furthermore, GAP, as a biopolymer matrix, was used to form the composite films with the small biological molecules cytidine-5′-monophosphate (CMP), adenosine-5′-monophosphate (AMP), and cyclic adenosine monophosphate (cAMP). Scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectrum, and X-ray diffraction (XRD) results showed that CMP, AMP, and cAMP interacted with the film substrates and might made films more complex. Notably, the addition of CMP, AMP, and cAMP promoted the light, water vapor, and oxygen barrier ability, surface wettability, mechanical strength, and antimicrobial activity against Gram-negative and –positive bacteria. Finally, GAP-based films composited with cAMP (cAMPF) exhibited the best characteristics were applied to fish packaging and preservation at 4 °C and extended the fish shelf life. All these data suggested the potential value of cAMPF as a functional edible polysaccharide film applied in food industries.
研究领域:
食品科学
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生物化学
·
海洋生物学
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产品关联
DOI
PubMed
高影响因子
CELL DEATH AND DIFFERENTIATION
·
2024
·
IF 13.7
Shi Dingfang,Bai Yunhao,Long Ruiling,Xia Jing,Xu Wenxuan,Qin Dongshen,Yang Xuejun,Ding Ming,Hou Xiao-Yu
Lysosomes regulate cellular metabolism to maintain cell survival, but the mechanisms whereby they determine neuronal cell fate after acute metabolic stress are unknown. Neuron-enriched lysosomal membrane protein LAMP2A is involved in selective chaperone-mediated autophagy and exosome loading. This study demonstrates that abnormalities in the neuronal LAMP2A-lysosomal pathway cause neurological deficits following ischemic stroke and that this is an early inducer of the PANoptosis-like molecular pathway and neuroinflammation, simultaneously inducing upregulation of FADD, RIPK3, and MLKL after ischemia. Quantitative proteomic and pharmacological analysis showed that after acute metabolic stress, the neuronal LAMP2A pathway induced acute synaptic degeneration and PANoptosis-like responses involving downregulation of protein kinase A (PKA) signaling. LAMP2A directed post-stroke lysosomal degradation of adenylyl cyclases (ADCY), including ADCY1 and ADCY3 in cortical neurons. Post-stroke treatment with cAMP mimetic or ADCY activator salvaged cortical neurons from PANoptosis-like responses and neuroinflammation, suggesting that the neuronal ADCY–cAMP–PKA axis is an upstream arrester of the pathophysiological process following an ischemic stroke. This study demonstrates that the neuronal LAMP2A-lysosmal pathway drives intricate acute neurodegenerative and neuroinflammatory responses after brain metabolic stress by downregulating the ADCY–PKA signaling cascade, and highlights the therapeutic potential of PKA signal inducers for improving stroke outcomes.
研究领域:
神经科学
·
细胞生物学
·
分子生物学
·
药理学
·
神经病学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2022
·Vol. 189
·期 4
·1-9
·
IF 6.408
Ye Tai,Zhu Dongdong,Hao Liling,Yuan Min,Cao Hui,Wu Xiuxiu,Yin Fengqin,Xu Fei
Kanamycin fluorescence aptasensors were created using a series of di-block oligonucleotide modified gold nanoparticles with various lengths of poly-adenine. In the presence of kanamycin, the double strand structure of the aptamer-reporter strand complex is disrupted, and the dye-labelled reporter strand detaches from the surface of gold nanoparticles, resulting in fluorescence recovery (Ex/Em = 485/520 nm). By adjusting the number of consecutive adenines, the programable aptamer density can be implemented on the gold nanoparticle surface, and the conformation of nucleic acid changed from lying-down to up-right. The apparent binding constant, binding kinetics, and limit of detection of the prepared aptasensors were carefully examined to explore the influence of surface density. Under the optimum condition, the aptasensor had a tenfold lower limit of detection than the thiolated aptamer modified one, as low as 23.6 nM, when a di-block oligonucleotide with twenty consecutive adenines tailed. In addition, satisfactory recoveries ranging from 96.33 to 99.47% were achieved in spiked milk samples with relative standard deviation of 1.2–6.9% ( n = 3). This surface density regulation strategy holds great promise in other aptamer-based interfacial recognition and sensing. Graphical abstract
Schematic presentation of di-block oligonucleotide modified gold nanoparticle with different surface densities and its kanamycin sensing application.
研究领域:
生物化学
·
纳米生物学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 6
·1-8
·
IF 5.833
Wang Hai-Bo,Mao An-Li,Tao Bei-Bei,Zhang Hong-Ding,Xiao Zhong-Liang,Liu Yan-Ming
An interesting phenomenon is described that the fluorescence signal of poly(adenine) (A) DNA-templated gold nanoclusters (AuNCs) is greatly improved in the presence of l -histidine by means of l -histidine-DNA interaction. The modified nanoclusters display strong fluorescence emission with excitation/emission maxima at 290/475 nm. The fluorescence quantum yield (QY) is improved from 1.9 to 6.5%. Fluorescence enhancement is mainly ascribed to the l -histidine-DNA interaction leading to conformational changes of the poly(A) DNA template, which offer a better microenvironment to protect AuNCs. The assay enables l -histidine to be determined with good sensitivity and a linear response that covers the 1 to 50 nM l -histidine concentration range with a 0.3 nM limit of detection. The proposed method has been applied to the determination of imidazole-containing drugs in pharmaceutical samples. Graphical abstract
A turn-on fluorescent method has been designed for the sensitive detection of l -histidine as well as imidazole-containing drugs on the basis of the l -histidine-DNA interaction.
研究领域:
纳米技术
·
分析化学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2018
·
IF 3.997
Qianqian Zhu,Wei Zhuang,Huanqing Niu,Lei Ge,Byron Villacorta Hernandez,Jinglan Wu,Kai Wang,Dong Liu,Yong Chen,Chenjie Zhu,Hanjie Ying
The development of an orientation immobilization technique via affinity between polyhistidine tags and metal ions aims at maintaining biocatalytic activity of the enzymes. In this work, to tackle the issue of the immobilization of adenylate cyclase (AC), a simple and effective approach of synthesizing iminodiacetic acid (IDA)-Ni 2+ particles was applied for simultaneously purifying and immobilizing his-tagged AC. We chose agarose particles as carriers, and then decorated them with IDA, leading to the formation of a coordination combination of Ni 2+ . The porous carriers with a large pore size of 50 nm and a specific surface area of 45.8 m 2 /g exhibited favorable enzymatic activity and loading capacity. The optimal pH of the immobilized enzyme increased from 8.0 to 9.0 and the optimal temperature increased from 30 °C to 35 °C, compared to the free AC. Moreover, the immobilized AC retained a residual activity of approximately 80% after storing it at 25 °C for 48 h, whereas only 40% of the activity was left in the free AC at the same conditions. Maximum yield of cyclic adenosine-3′, 5′- monophosphate (cAMP) reached up to the summit of the reaction. The immobilized AC by affinity adsorption will provide a promising route for the industrial production of cAMP.
研究领域:
生物化学
·
酶工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2016
·Vol. 39
·期 8
·1533-1541
·
IF 2.741
Xuan Guo,Tingting Ye,Luying Liu,Xiaogang Hu
An aptamer with adenosine triphosphate as a ligand was immobilized onto the surface of a porous-polymer-coated fiber to obtain an aptamer-functionalized porous-polymer-coated solid-phase microextraction fiber. The fiber was observed with a crosslinked and porous morphological surface structure. It shows specific selectivity to adenosine triphosphate with a selectivity coefficient of 22.1 compared to the scrambled oligonucleotide functionalized fiber, and the selectivity factors over other analogues and reference compounds were from 6.1 to 77.5. When the fiber-based solid-phase microextraction was coupled with liquid chromatography and tandem mass spectrometry, detection limits of 2.7, 29, and 34 μg/L were achieved for adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate, respectively. The spiking recoveries of 77.6–91.9% were achieved for trace adenosine phosphates in human serum sample. Furthermore, the fibers showed high stability and good reusability and could be used over 50 times for the real serum sample pretreatment without remarkable performance reduction.
研究领域:
分析化学
·
生物分析
✓ 已验证
DOI
PubMed
高影响因子
CHEMBIOCHEM
·
2024
·
IF 2.6
Mengyuan Yin,Xiaohong Hu,Yu Chen,Hanxue Liang,Yuxin Shen,Weiwei Guo
Smart shape-memory DNA hydrogels, which can respond to various types of external stimuli and undergo macroscopic shape deformations, have shown great potential in various applications. By constructing free-standing films, the deformation and response properties of these hydrogels can be further enhanced, and visualized deformation can be achieved. However, DNA hydrogels that can exhibit rapid and high-degree shape deformations, such as the inverse shape deformations, are still lacking. Herein, free-standing oligoadenine strand-functionalized polyacrylamide hydrogel films were developed that can exhibit rapid and high-degree of inverse shape deformation upon cyclic pH changes. The oligoadenine strands exhibit a pH-stimulated reversible conformational transition between a flexible single-stranded state and parallel duplex A-motif structures, resulting in their role change in the film from negatively charged side chains to “head-to-head” crosslinking structures, driving a high degree of inverse shape deformation with a relative bending angle change of 223.7%, which is more than 5 times that of a film driven by pH-responsive i-motif structures, facilitating the development of bilayer hydrogel film actuators with potential in flexible sensors and robots.
研究领域:
生物材料学
·
高分子化学
·
智能材料
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产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2019
·Vol. 24
·期 4
·663
·
IF 3.06
Kai Song,Yejing Wang,Yu Li,Chaoxiang Ding,Rui Cai,Gang Tao,Ping Zhao,Qingyou Xia,Huawei He
Enzymatic activity assays are essential and critical for the study of enzyme kinetics. Adenylate kinase (Adk) plays a fundamental role in cellular energy and nucleotide homeostasis. To date, assays based on different principles have been used for the determination of Adk activity. Here, we show a spectrophotometric analysis technique to determine Adk activity with bromothymol blue as a pH indicator. We analyzed the effects of substrates and the pH indicator on the assay using orthogonal design and then established the most optimal assay for Adk activity. Subsequently, we evaluated the thermostability of Adk and the inhibitory effect of KCl on Adk activity with this assay. Our results show that this assay is simple, rapid, and precise. It shows great potential as an alternative to the conventional Adk activity assay. Our results also suggest that orthogonal design is an effective approach, which is very suitable for the optimization of complex enzyme reaction conditions.Keywords:enzymatic activity assay;adenylate kinase;spectrophotometry;orthogonal experiment;bromothymol blue
研究领域:
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）