Biomedical Engineering Online
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2019
·Vol. 18
·期 1
·1-16
·
IF 2.013
Fang Ao,Li Dichen,Hao Zhiyan,Wang Ling,Pan Binglei,Gao Lin,Qu Xiaoli,He Jiankang
Background Human brain models and pharmacological models of brain diseases are in high demand for drug screening because animal models have been found to be less than ideal for fully representing the human brain and are likely to fail during drug screening and testing; therefore, the construction of brain-like tissues is necessary. Due to the complexity of cortical tissue, the in vitro construction of brain-like tissue models has been restricted to mostly two-dimensional (2D) models and, on a limited scale, three-dimensional (3D) models.Methods In this study, 3D tissue blocks encapsulating neurons and astrocytes were constructed and cultured in vitro to mimic the cortex of the brain and to investigate the effects of astrocytes on the growth of neurons in a 3D culture.Results The results indicated that such methodology can provide a 3D culture environment suitable for neurons and astrocytes to live and function. When both cells were evenly mixed and cultured in a 3D manner, the astrocytes, which showed better outgrowth and a higher proliferation rate, benefited more than the neurons. On the other hand, the neurons benefited, showing longer axons and a denser network of dendrites, when they were accompanied by astrocytes at a certain distance.Conclusion In conclusion, astrocytes stimulated the outgrowth of neurons in a 3D culture environment in vitro. Regardless, the spatial relationship between both types of cells should be controlled. Thus, culturing cells in a 3D manner is necessary to investigate the correlations between them. This study provides a foundation for biofabricating 3D neurons’ cultures to allow for a deeper insight into the relationship between astrocytes or other glial cells and neurons in a 3D culture that is similar to the natural environment of the brain.
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Nature Communications
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2019
·Vol. 10
·期 1
·1-9
·
IF 11.878
Zhu Yujiao,Huang Ziyu,Chen Qingming,Wu Qian,Huang Xiaowen,So Pui-Kin,Shao Liyang,Yao Zhongping,Jia Yanwei,Li Zhaohui,Yu Weixing,Yang Yi,Jian Aoqun,Sang Shengbo,Zhang Wendong,Zhang Xuming
Food production in green crops is severely limited by low activity and poor specificity of D-ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) in natural photosynthesis (NPS). This work presents a scientific solution to overcome this problem by immobilizing RuBisCO into a microfluidic reactor, which demonstrates a continuous production of glucose precursor at 13.8 μmol g −1 RuBisCO min −1 from CO 2 and ribulose-1,5-bisphosphate. Experiments show that the RuBisCO immobilization significantly enhances enzyme stabilities (7.2 folds in storage stability, 6.7 folds in thermal stability), and also improves the reusability (90.4% activity retained after 5 cycles of reuse and 78.5% after 10 cycles). This work mimics the NPS pathway with scalable microreactors for continuous synthesis of glucose precursor using very small amount of RuBisCO. Although still far from industrial production, this work demonstrates artificial synthesis of basic food materials by replicating the light-independent reactions of NPS, which may hold the key to food crisis relief and future space colonization.
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Biotechnology for Biofuels
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2013
·Vol. 6
·期 1
·1-10
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IF 5.552
Yang Jianming,Nie Qingjuan,Ren Meng,Feng Hongru,Jiang Xinglin,Zheng Yanning,Liu Min,Zhang Haibo,Xian Mo
Background α-Pinene is an important natural product that is widely used in flavorings, fragrances, medicines, fine chemicals and high-density renewable fuels. Currently, α-Pinene used in industry is mainly produced either by tapping trees (gum turpentine) or as a byproduct of paper pulping (crude sulfate turpentine, CST). However, the extraction of it from trees is tedious and inefficient and requires substantial expenditure of natural resources. Therefore, it is necessary to seek sustainable technologies for α-pinene production.Results To construct the microbial synthetic pathway of α-pinene in E. coli, we co-expressed native geranyl diphosphate synthase (IspA) from E. coli and α-pinene synthase (Pt30) from Pinus taeda, and then to increase the geranyl diphosphate (GPP) content in the cells, a suitable geranyl diphosphate synthase (GPPS2) was selected from two different origins. Furthermore, to enhance α-pinene production, a novel biosynthetic pathway of α-pinene was assembled in E. coli BL21(DE3) with the heterologous hybrid mevalonate (MVA) pathway, GPPS2 and α-pinene synthase (Pt30). The final genetic strain, YJM28, harboring the above novel biosynthetic pathway of α-pinene, accumulated α-pinene up to 5.44 mg/L and 0.97 g/L under flask and fed-batch fermentation conditions, respectively. The conversion efficiency of glucose to α-pinene (gram to gram) in the metabolically engineered strain reached 2.61%.Conclusion sIn this paper, by using metabolic engineering techniques, the more efficient biosynthetic pathway of α-pinene was successfully assembled in E. coli BL21(DE3) with the heterologous hybrid MVA pathway, GPPS2 and α-pinene synthase (Pt30). In addition, this is the first report on α-pinene fed-batch fermentation, and our results represent improvements over previous reports.
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AMB Express
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2022
·Vol. 12
·期 1
·1-17
·
IF 4.126
Liu Shasha,She Pengfei,Li Zehao,Li Yimin,Li Linhui,Yang Yifan,Zhou Linying,Wu Yong
High incidences of urinary tract infection (UTI) of aminoglycosides-resistant E.coli causes a severe burden for public health. A new therapeutic strategy to ease this crisis is to repurpose non-antibacterial compounds to increase aminoglycosides sensibility against multidrug resistant E.coli pathogens. Based on high throughput screening technology, we profile the antimicrobial activity of tavaborole, a first antifungal benzoxaborole drug for onychomycosis treatment, and investigate the synergistic interaction between tavaborole and aminoglycosides, especially tobramycin and amikacin. Most importantly, by resistance accumulation assay, we found that, tavaborole not only slowed resistance occurrence of aminoglycosides, but also reduced invasiveness of E.coli in combination with tobramycin. Mechanistic studies preliminary explored that tavaborole and aminoglycosides lead to mistranslation, but would be still necessary to investigate more details for further research. In addition, tavaborole exhibited low systematic toxicity in vitro and in vivo, and enhanced aminoglycoside bactericidal activity in mice peritonitis model. Collectively, these results suggest the potential of tavaborole as a novel aminoglycosides adjuvant to tackle the clinically relevant drug resistant E. coli and encourages us to discover more benzoxaborole analogues for circumvention of recalcitrant infections.
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Reaction Kinetics Mechanisms and Catalysis
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2026
·
IF 2.1
Liu Kun,Shu Anjie,Zhao Liang,Qiu Bo,Wang Jiefeng
Herein, a synergistic photothermal-photocatalytic strategy is developed by designing a contactless photocatalytic reactor that concurrently synchronizes and enhances mass transfer while expanding the scope of usable water sources. Specifically, heat radiation-induced evaporation is exploited to accelerate the transport of both reactants and products, thereby relaxing the purity requirements for the water feedstock and sustaining efficient CO 2 reduction rates even with seawater and wastewater. Importantly, the contactless gas-phase configuration is expected to reduce the mass-transfer mismatch between CO 2 and H 2 O by bringing both reactants into the gas phase, where their diffusion coefficients are of the same order of magnitude. Under full-spectrum irradiation in a reactor with a 1 cm 2 irradiation area, the CO 2 -to-formic acid production rates using pure water, simulated seawater, and simulated wastewater reach 680.7, 645.3, and 663.0 μmol g −1 h −1 . This work not only demonstrates the valorization of wastewater resources but also opens a new avenue for integrated photocatalysis and photothermal conversion.
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MICROCHIMICA ACTA
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2018
·Vol. 185
·期 9
·1-9
·
IF 5.705
Gou Dan,Xie Guoming,Li Yuxia,Zhang Xin,Chen Hui
The authors describe an electrochemical immunoassay for ultrasensitive determination of the Mycobacterium tuberculosis (MTb) secretory protein MPT64 which is an antigen for early diagnosis of infection with MTb . Protein G was used to immobilize antibodies against MPT64 on a gold electrode. Graphene oxide with its large surface area was used as a carrier to anchor magnetite (Fe 3 O 4 ) and platinum (Pt) nanoparticles. The nanocomposite of type GO@Fe 3 O 4 @Pt was used as a signal reporter with excellent catalytic activity and recyclability. The nanocomposite exhibits peroxidase-like activity for hydrogen peroxide, best at −0.25 V (vs. Ag/AgCl). It works over a wide range of pH values (2–10) and temperatures (25–65 °C). Further signal amplification strategy was accomplished by rolling circle amplification. This voltammetric assay has a linear response to the logarithm of MPT64 concentration in the range from 5.0 fg·mL −1 to 1.0 ng·mL −1 and a detection limit of 0.34 fg·mL −1 (at a signal to noise ratio of 3, for n = 10). The assay can be completed within 4 h. It was successfully applied to the determination of MPT64 in spiked serum samples. Conceivably, the assay has a large potential in providing laboratory evidence for rapid diagnosis of MTb infection.
An electrochemical biosensor was developed for rapid detection of Mycobacterium tuberculosis secretory protein MPT64. The detection signal was synergistically amplified by GO@Fe 3 O 4 @Pt and rolling circle amplification. Phi29: phi29 DNA polymerase; BSA: bovine serum albumin; GO: graphene oxide.
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MICROCHIMICA ACTA
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2020
·Vol. 187
·期 8
·1-8
·
IF 6.232
Chang Shuai,Chen Bin Bin,Lv Jian,Fodjo Essy Kouadio,Qian Ruo Can,Li Da Wei
Lysosomes with a single-layered membrane structure are mainly involved in the scavenging of foreign substances and play an important role in maintaining normal physiological functions of living cells. In this work, near-neutrally charged fluorescent carbon dots (CDs) were prepared with lipophilicity through a facile one-pot hydrothermal carbonization of chloranil and triethylenetetramine at 160 °C for 3 h. The as-obtained CDs are proved to have good photostability, low cost, and excellent biocompatibility. Importantly, the as-prepared CDs with high quantum yield of 30.8% show excitation-dependent emission with great stability, and thus, they can be well used for the long-term target imaging of lysosomes in living cells without further modification. Meanwhile, the CDs can quickly enter into the lysosomes within 30 min, and the green fluorescence (FL) of CDs reaches the plateau when incubated for 60 min. By comparing the fluorescent intensity, the information about distribution and amount of lysosomes in different cells can be obtained. The proposed CD-based strategy demonstrates great promise for label-free target imaging of lysosomes in living cells.
The near-neutral carbon dots (CDs) with lipophilicity are used as label-free fluorescent nanoprobes for the long-term imaging of lysosomes in living cells.
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JOURNAL OF NANOBIOTECHNOLOGY
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2022
·Vol. 20
·期 1
·1-19
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IF 9.429
Peng Xin,Xie Zhuomi,Wang Xiuhua,Zhao Yuxiang,Yang Chuyun,Zhang Zhongyi,Li Mingjie,Zheng Jianping,Wang Yuhui
Background Luminescent nanomaterials (LNMs), especially newly-exploited fluorescent carbon-dots (CDs), have demonstrated promising candidates for sunlight harvesting and enhanced photosynthesis efficiency of crops. However, most of the studies focus on the design and synthesis of LNMs and primary metabolism in biomass acceleration, secondary metabolism that closely associated with the quality ingredients of plants is rarely mentioned. Results UV-absorptive and water-soluble NIR-CDs were harvested via a facile microwave-assisted carbonization method. The effect and regulatory mechanism of NIR-CDs on the secondary metabolism and bioactive ingredients accumulation in Tetrastigma hemsleyanum were explored. A total of 191 differential secondary metabolites and 6874 differentially expressed genes were identified when the NIR-CDs were adopted for enhancing growth of T. hemsleyanum . The phenolic acids were generally improved, but the flavonoids were more likely to decrease. The pivotal differentially expressed genes were involved in biosynthesis of secondary metabolites, flavonoid biosynthesis, porphyrin and chlorophyll metabolism, etc. The gene-metabolite association was constructed and 44 hub genes highly related to quality ingredients accumulation and growth were identified, among which and the top 5 genes of the PPI network might be the key regulators. Conclusion This research provided key regulatory genes and differentially accumulating quality ingredients under NIR-CDs-treatment, which could provide a theoretical basis for expanding the applications of nanomaterial in industrial crop agriculture. Graphical Abstract
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MICROCHIMICA ACTA
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2022
·Vol. 189
·期 10
·1-13
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IF 6.408
Li Xinhao,Li Shengnan,Lv Qiuyan,Wang Chaoxian,Liang Jintao,Zhou Zhide,Li Guiyin
A Golgi protein 73 (GP73) colorimetric biosensor based on the reduced graphene oxide-carboxymethyl chitosan-hemin/platinum@palladium nanoparticles (RGO-CMCS-Hemin/Pt@Pd NPs) with peroxidase-like activity was constructed. The RGO-CMCS-Hemin/Pt@Pd NPs with high peroxidase-like activity were successfully synthesized under mild conditions. Then, the aminylated GP73 aptamer (Apt) was bound to the RGO-CMCS-Hemin/Pt@Pd NPs to form the recognition probe. Another unmodified GP73 aptamer (AptI) was served as the capture probe. In the presence of target GP73, the capture probe and the recognition probe specifically bind to GP73 and form a RGO-CMCS-Hemin/Pt@Pd NP-Apt/GP73/AptI sandwich-type structure, which can oxidase the colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxTMB in the presence of H 2 O 2 . GP73 detection was achieved by measuring the peak UV absorption at 652 nm. Under the optimum conditions, the GP73 concentration was linearly related to the absorbance intensity in the range 10.0–110.0 ng/mL, and the limit of detection (LOD) was 4.7 ng/mL. The proposed colorimetric biosensor was successfully applied to detect GP73 in spiked human serum samples with recoveries of 98.2–107.0% and RSDs of 1.90–5.44%, demonstrating the excellent potential for highly sensitive GP73 detection in clinical detection. Graphical abstract A colorimetric biosensor for visual determination of GP73 based on RGO-CMCS-Hemin/Pt@Pd NPs nanozyme with peroxidase-like activity was designed. The GP73 biosensor responses linearly from 10.0–110.0 ng/mL with LOD of 4.7 ng/mL, and shows acceptable specificity and good recovery.
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MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 4
·1-8
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IF 5.833
Wu Daohong,He Yuhan,Tong Liujuan,Wang Jianxiu,Liu Lin,Yi Xinyao,Hu Shengqiang
An electrochemical sensor capable of quantitative determination of caspase-3 activities was developed. A thiolated peptide whose sequence contained a caspase-3 cleaved site and a cell penetration sequence was preimmobilized onto an electrode. The quantification of caspase-3 was accomplished after cell penetration and the subsequent adsorption of silver nanoparticles (AgNPs). The oxidation current of AgNPs was found to be inversely proportional to the concentration of caspase-3 between 0.02 and 0.2 U/mL. A detection limit of 0.02 U/mL for caspase-3 was achieved due to the large number of positively charged AgNPs adsorbed onto the negatively charged cells. The proof of concept was demonstrated by monitoring the cleavage of surface-confined peptide substrates by caspase-3 in cell lysates. The current sensor could be extended to detect cells by replacing the surface-confined peptide with aptamers that recognize cells. Thus, the use of a cell as a matrix for AgNPs shows excellent potential for constructing electrochemical sensors and provides a useful alternative for sensor development in the future. Graphical abstract
Cells modified with silver nanoparticles were utilized as the electrochemical readout of an electrochemical assay.
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Nature Protocols
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2021
·Vol. 16
·期 9
·4201-4226
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IF 13.491
Gao Pengcheng,Wang Dagui,Che Cheng,Ma Qun,Wu Xiaoqing,Chen Yajie,Xu Hongquan,Li Xinchun,Lin Yu,Ding Defang,Lou Xiaoding,Xia Fan
Solid-state nanochannels (SSNs) provide a promising approach for biosensing due to the confinement of molecules inside, their great mechanical strength and diversified surface chemical properties; however, until now, their sensitivity and specificity have not satisfied the practical requirements of sensing applications, especially in complex matrices, i.e., media of diverse constitutions. Here, we report a protocol to achieve explicit regional and functional division of functional elements at the outer surface (FE OS ) and inner wall (FE IW ) of SSNs, which offers a nanochannel-based sensing platform with enhanced specificity and sensitivity. The protocol starts with the fabrication and characterization of the distribution of FE OS and FE IW . Then, the evaluation of the contributions of FE OS and FE IW to ionic gating is described; the FE IW mainly regulate ionic gating, and the FE OS can produce a synergistic effect. Finally, hydrophobic or highly charged FE OS are applied to ward off interference molecules, non-target molecules that may affect the ionic signal of nanochannels, which decreases false signals and helps to achieve the highly specific ionic output in complex matrices. Compared with other methods currently available, this method will contribute to the fundamental understanding of substance transport in SSNs and provide high specificity and sensitivity in SSN-based analyses. The procedure takes 3–6 d to complete.
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CELLULOSE
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2022
·Vol. 29
·期 4
·2479-2495
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IF 6.123
Tang Ruihua,Li Min,Yan Xueyan,Xie Mingyue,Liu Li Na,Li Zedong,Xu Feng
Cheap, rapid, simple and equipment-free nucleic acid extraction (NAE) is highly preferred for implementing nucleic acid detection at point-of-care (POC). Paper-based NAE materials have been extensively utilized due to their low cost, abundance, portability, biocompatibility and ease of chemical modification. However, it is challenging for users to choose the proper one from existing paper-based NAE materials for specific POC applications, which is determined by their physical and chemical properties. Additionally, building the relationship between the physical and chemical properties and the NAE efficiency of paper-based materials is instructive for development of new paper-based NAE materials. In this study, we first systematically compared the physical and chemical properties of six widely used paper-based NAE materials (namely Whatman filter paper #1, FTA card, FTA elute card, Fusion 5, silica membrane and polyethersulfone (PES) membrane), and then evaluated their NAE efficiency. The obtained results indicated that pore uniformity, wet strength, porosity and functional groups are key parameters to affect the efficiency of NAE. The NAE performance of FTA card is the best with high concentration and purity. Finally, we envision that more cost-effective paper-based NAE materials will be developed for POCT application in the future. Graphic abstract
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危险品化学品经营许可证（带存储）