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学者姓名：陈晓超
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Abstract ：
The associative phase separation of charged biomacromolecules plays a key role in many biophysical events that take place in crowded intracellular environments. Such natural polyelectrolyte complexation and phase separation often occur at nonstoichiometric charge ratios with the incorporation of bioactive proteins, which is not studied as extensively as those complexations at stoichiometric ratios. In this work, we investigated how the addition of a crowding agent (polyethylene glycol, PEG) affected the complexation between chitosan (CS) and hyaluronic acid (HA), especially at nonstoichiometric ratios, and the encapsulation of enzyme (catalase, CAT) by the colloidal complexes. The crowded environment promoted colloidal phase separation at low charge ratios, forming complexes with increased colloidal and dissolution stability, which resulted in a smaller size and polydispersity (PDI). The binding isotherms revealed that the addition of PEG greatly enhanced the ion-pairing strength (with increased ion-pairing equilibrium constant K-a from 4.92 x 10(4) without PEG to 1.08 x 10(6) with 200 g/L PEG) and switched the coacervation from endothermic to exothermic, which explained the promoted complexation and phase separation. At the stoichiometric charge ratio, the enhanced CS-HA interaction in crowded media generated a more solid-like coacervate phase with a denser network, slower chain relaxation, and higher modulus. Moreover, both crowding and complex encapsulation enhanced the activity and catalytic efficiency of CAT, represented by a 2-fold increase in catalytic efficiency (K-cat/K-m) under 100 g/L PEG crowding and CS-HA complex encapsulation. This is likely due to the lower polarity in the microenvironment surrounding the enzyme molecules. By a systematic investigation of both nonstoichiometric and stoichiometric charge ratios under macromolecular crowding, this work provided new insights into the complexation between natural polyelectrolytes in a scenario closer to an intracellular environment.
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| GB/T 7714 | Ou, Xiatong , Tang, Ziyao , Ye, Yanqi et al. Macromolecular Crowding Effect on Chitosan-Hyaluronic Acid Complexation and the Activity of Encapsulated Catalase [J]. | BIOMACROMOLECULES , 2024 , 25 (6) : 3840-3849 . |
| MLA | Ou, Xiatong et al. "Macromolecular Crowding Effect on Chitosan-Hyaluronic Acid Complexation and the Activity of Encapsulated Catalase" . | BIOMACROMOLECULES 25 . 6 (2024) : 3840-3849 . |
| APA | Ou, Xiatong , Tang, Ziyao , Ye, Yanqi , Chen, Xiaochao , Huang, Yan . Macromolecular Crowding Effect on Chitosan-Hyaluronic Acid Complexation and the Activity of Encapsulated Catalase . | BIOMACROMOLECULES , 2024 , 25 (6) , 3840-3849 . |
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目的：观察三七总皂苷(PNS)结合细胞穿透肽Tat(47-57)对骨折的愈合作用，并探讨其可能机制。方法：研究选取成年雄性SD大鼠建立骨折实验动物模型，实验动物随机分为四组：空白对照组、外用三七总皂苷组(PNS外用组)、外用三七总皂苷和细胞穿透肽联合组(Tat-PNS外用组)、口服三七总皂苷组(PNS口服组)。给药14d后，检测骨折病灶局部的抗氧化SOD酶活性，同时取骨折病灶局部进行病理切片分析，对比和分析各给药组对实验大鼠骨折愈合过程的影响程度。结果：与空白对照组对比，PNS外用组对大鼠骨折愈合无明显的疗效，可能与PNS无法有效透皮吸收，并在病灶局部富集有效浓度有关。Tat-PNS外用组能够...
Keyword ：
SOD SOD 三七总皂苷 三七总皂苷 抗氧化 抗氧化 组织切片 组织切片 细胞穿透肽 细胞穿透肽 骨形态发生蛋白-2 骨形态发生蛋白-2 骨折 骨折
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| GB/T 7714 | 蒋擎 , 陈晓超 , 田康勇 et al. 三七总皂苷结合细胞穿透肽Tat经皮给药促进骨折愈合的研究 [J]. | 医学理论与实践 , 2022 , 35 (10) : 1621-1623,1627 . |
| MLA | 蒋擎 et al. "三七总皂苷结合细胞穿透肽Tat经皮给药促进骨折愈合的研究" . | 医学理论与实践 35 . 10 (2022) : 1621-1623,1627 . |
| APA | 蒋擎 , 陈晓超 , 田康勇 , 谢超春 , 刘腾鸿 , 林燕云 et al. 三七总皂苷结合细胞穿透肽Tat经皮给药促进骨折愈合的研究 . | 医学理论与实践 , 2022 , 35 (10) , 1621-1623,1627 . |
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目的:去除重组毕赤酵母表达人Cu,Zn-SOD蛋白溶液中的内毒素,为后续活体动物试验和临床应用提供基础数据。方法:采用Triton X-114萃取与活性炭吸附两种方法单独使用或联合使用的纯化方案,比较不同纯化方案对去除重组蛋白溶液中内毒素以及目标蛋白回收率的效果。结果:通过单因素条件优化,确定重组蛋白溶液最佳操作pH为5.0,活性炭最适添加量为5%以及Triton X-114最适添加量为1%(体积分数)。最后确定活性炭吸附和Triton X-114萃取法联合使用的三步法操作工艺,可高效去除重组蛋白溶液中内毒素。终产品内毒素含量降至26.8EU/10 000 U SOD,去除率达到99.8%,目...
Keyword ：
Triton X-114 Triton X-114 内毒素 内毒素 毕赤酵母 毕赤酵母 活性炭 活性炭 蛋白溶液 蛋白溶液 超氧化物歧化酶 超氧化物歧化酶 重组蛋白 重组蛋白
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| GB/T 7714 | 陈晓超 , 王友 , 崔文明 et al. 重组SOD发酵液去除内毒素的效果研究 [J]. | 中国食品学报 , 2019 , 19 (09) : 172-178 . |
| MLA | 陈晓超 et al. "重组SOD发酵液去除内毒素的效果研究" . | 中国食品学报 19 . 09 (2019) : 172-178 . |
| APA | 陈晓超 , 王友 , 崔文明 , 李伟松 , 刘树滔 , 饶平凡 . 重组SOD发酵液去除内毒素的效果研究 . | 中国食品学报 , 2019 , 19 (09) , 172-178 . |
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Abstract ：
目的:去除重组毕赤酵母表达人Cu,Zn-SOD蛋白溶液中的内毒素,为后续活体动物试验和临床应用提供基础数据.方法:采用Triton X-114萃取与活性炭吸附两种方法单独使用或联合使用的纯化方案,比较不同纯化方案对去除重组蛋白溶液中内毒素以及目标蛋白回收率的效果.结果:通过单因素条件优化,确定重组蛋白溶液最佳操作pH为5.0,活性炭最适添加量为5％以及Triton X-114最适添加量为1％(体积分数).最后确定活性炭吸附和Triton X-114萃取法联合使用的三步法操作工艺,可高效去除重组蛋白溶液中内毒素.终产品内毒素含量降至26.8EU/10000 U SOD,去除率达到99.8％,目标蛋白回收率为66％,Triton X-114残余量为5.7μg/mL.结论:纯化后所得重组SOD蛋白溶液符合中国药典规定的注射用针剂中内毒素限量与Triton残余量的质量要求,可用于潜在的保健食品开发.
Keyword ：
Triton X-114 Triton X-114 内毒素 内毒素 毕赤酵母 毕赤酵母 活性炭 活性炭 蛋白溶液 蛋白溶液 超氧化物歧化酶 超氧化物歧化酶 重组蛋白 重组蛋白
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| GB/T 7714 | 陈晓超 , 王友 , 崔文明 et al. 重组SOD发酵液去除内毒素的效果研究 [J]. | 中国食品学报 , 2019 , 19 (9) : 172-178 . |
| MLA | 陈晓超 et al. "重组SOD发酵液去除内毒素的效果研究" . | 中国食品学报 19 . 9 (2019) : 172-178 . |
| APA | 陈晓超 , 王友 , 崔文明 , 李伟松 , 刘树滔 , 饶平凡 . 重组SOD发酵液去除内毒素的效果研究 . | 中国食品学报 , 2019 , 19 (9) , 172-178 . |
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The aim of this study was to investigate whether the cellular uptake of cargo proteins can be enhanced by fusing a Tat peptide in the center of proteins; glutathione-S-transferase (GST)-Tat-green fluorescent protein (GFP) and GST-GFP-Tat proteins were first constructed and expressed. The cellular internalization of both proteins was then evaluated and compared in HeLa cells using fluorescent microscopy and flow cytometry, as well as the transdermal delivery in human skin using confocal microscopy. Results from in vitro cell experiments showed that GST-Tat-GFP protein efficiently internalized into HeLa cells when a Tat peptide was fused in the center of proteins, whereas its efficiency is lower than that of GST-GFP-Tat protein with a Tat peptide terminal fused. Ex vivo transdermal delivery data also demonstrated that the lower efficiency of GST-Tat-GFP penetrating through human stratum corneum layer when compared with GST-GFP-Tat. Furthermore, both GST-GFP-Tat and GST-Tat-GFP presented a various degree of a mixture of cytoplasmic diffuse staining and punctate surface staining, and the pattern of distribution varied considerably in HeLa cell experiments depending on the concentration of protein used. Therefore, an improved mechanism for Tat-conjugated proteins was proposed, in which Tat-conjugated proteins were first associated with cell membrane, then accumulated on the cell surface, and finally internalized into cells by pore formation mechanism. (c) 2018 American Pharmacists Association (R). Published by Elsevier Inc. All rights reserved.
Keyword ：
cell-penetrating peptide cell-penetrating peptide cellular internalization cellular internalization confocal microscopy confocal microscopy fluorescence microscopy fluorescence microscopy green fluorescent protein green fluorescent protein Tat peptide Tat peptide transdermal delivery transdermal delivery
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| GB/T 7714 | Chen, Xiaochao , Chen, Jing , Fu, Rong et al. Can the Cellular Internalization of Cargo Proteins Be Enhanced by Fusing a Tat Peptide in the Center of Proteins? A Fluorescence Study [J]. | JOURNAL OF PHARMACEUTICAL SCIENCES , 2018 , 107 (3) : 879-886 . |
| MLA | Chen, Xiaochao et al. "Can the Cellular Internalization of Cargo Proteins Be Enhanced by Fusing a Tat Peptide in the Center of Proteins? A Fluorescence Study" . | JOURNAL OF PHARMACEUTICAL SCIENCES 107 . 3 (2018) : 879-886 . |
| APA | Chen, Xiaochao , Chen, Jing , Fu, Rong , Rao, Pingfan , Weller, Richard , Bradshaw, Jeremy et al. Can the Cellular Internalization of Cargo Proteins Be Enhanced by Fusing a Tat Peptide in the Center of Proteins? A Fluorescence Study . | JOURNAL OF PHARMACEUTICAL SCIENCES , 2018 , 107 (3) , 879-886 . |
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胶原蛋白的抗氧化活性尚未见报道,而其水解产物胶原蛋白肽的抗氧化活性报道较多。为研究造成该差异性的原因,分别采用T-AOC法、盐酸羟胺法、DPPH法和ABTS法测定胶原蛋白模拟肽、胶原蛋白肽混合物及其组成氨基酸脯氨酸和甘氨酸的抗氧化活性,并用圆二色光谱法测定CMP和CP的二级结构。试验结果表明:(1)T-AOC法中,CMP,CP和各种氨基酸都不具有抗氧化活性;(2)在盐酸羟胺法中,只有CP具有抗氧化活性;(3)在DPPH法和ABTS法中,肽和氨基酸都具有一定的抗氧化活性,且抗氧化顺序为CP>Hypro>CMP≥Pro>Gly;(4)圆二色光谱法结果表明,CMP具有胶原蛋白形成稳定的3股螺旋结构所...
Keyword ：
3股螺旋结构 3股螺旋结构 抗氧化 抗氧化 胶原蛋白 胶原蛋白 胶原蛋白肽 胶原蛋白肽
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| GB/T 7714 | 林春通 , 柯欣欣 , 郭建辉 et al. 胶原蛋白与胶原蛋白肽的抗氧化关系初探 [J]. | 中国食品学报 , 2017 , 17 (10) : 44-50 . |
| MLA | 林春通 et al. "胶原蛋白与胶原蛋白肽的抗氧化关系初探" . | 中国食品学报 17 . 10 (2017) : 44-50 . |
| APA | 林春通 , 柯欣欣 , 郭建辉 , 巫启明 , 刘树滔 , 郭静科 et al. 胶原蛋白与胶原蛋白肽的抗氧化关系初探 . | 中国食品学报 , 2017 , 17 (10) , 44-50 . |
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The attractive potential of natural superoxide dismutase (SOD) in the fields of medicine and functional food is limited by its short half-life in circulation and poor permeability across the cell membrane. The nanoparticle form of SOD might overcome these limitations. However, most preparative methods have disadvantages, such as complicated operation, a variety of reagentssome of them even highly toxicand low encapsulation efficiency or low release rate. The aim of this study is to present a simple and green approach for the preparation of SOD nanoparticles (NPs) by means of co-incubation of Cu/Zn SOD with glucose. This method was designed to prepare nanoscale aggregates based on the possible inhibitory effect of Maillard reaction on heating-induced aggregation during the co-incubation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results indicated that the Maillard reaction occurred during the co-incubation process. It was found that enzymatically active NPs of Cu/Zn SOD were simultaneously generated during the reaction, with an average particle size of 175.86 +/- 0.71 nm, and a Zeta potential of -17.27 +/- 0.59 mV, as established by the measurement of enzymatic activity, observations using field emission scanning electron microscope, and analysis of dynamic light scattering, respectively. The preparative conditions for the SOD NPs were optimized by response surface design to increase SOD activity 20.43 fold. These SOD NPs showed storage stability for 25 days and better cell uptake efficacy than natural SOD. Therefore, these NPs of SOD are expected to be a potential drug candidate or functional food factor. To our knowledge, this is the first report on the preparation of nanoparticles possessing the bioactivity of the graft component protein, using the simple and green approach of co-incubation with glucose, which occurs frequently in the food industry during thermal processing.
Keyword ：
cell uptake cell uptake Maillard reaction Maillard reaction nanoparticle nanoparticle superoxide dismutase superoxide dismutase
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| GB/T 7714 | Cai, Liping , Lin, Chuntong , Yang, Nannan et al. Preparation and Characterization of Nanoparticles Made from Co-Incubation of SOD and Glucose [J]. | NANOMATERIALS , 2017 , 7 (12) . |
| MLA | Cai, Liping et al. "Preparation and Characterization of Nanoparticles Made from Co-Incubation of SOD and Glucose" . | NANOMATERIALS 7 . 12 (2017) . |
| APA | Cai, Liping , Lin, Chuntong , Yang, Nannan , Huang, Zhijie , Miao, Song , Chen, Xiaochao et al. Preparation and Characterization of Nanoparticles Made from Co-Incubation of SOD and Glucose . | NANOMATERIALS , 2017 , 7 (12) . |
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The aim of this study is to investigate the interactions between TAT peptides and a neutral DOPC bilayer by using neutron lamellar diffraction. The distribution of TAT peptides and the perturbation of water distribution across the DOPC bilayer were revealed. When compared to our previous study on an anionic DOPC/DOPS bilayer (X. Chen et al., Biochim Biophys Acta. 2013. 1828 (8), 1982-1988), a much deeper insertion of TAT peptides was found in the hydrophobic core of DOPC bilayer at a depth of 6.0 A from the center of the bilayer, a position close to the double bond of fatty acyl chain. We conclude that the electrostatic attractions between the positively charged TAT peptides and the negatively charged headgroups of phospholipid are not essential for the direct translocation. Furthermore, the interactions of TAT peptides with the DOPC bilayer were found to vary in a concentration -dependent manner. A limited number of peptides first associate with the phosphate moieties on the lipid headgroups by using the guanidinium ions pairing. Then the energetically favorable water defect structures are adopted to maintain the arginine residues hydrated by drawing water molecules and lipid headgroups into the bilayer core. Such bilayer deformations consequently lead to the deep intercalation of TAT peptides into the bilayer core. Once a threshold concentration of TAT peptide in the bilayer is reached, a significant rearrangement of bilayer will happen and steady-state water pores will form. (C) 2017 Elsevier B.V. All rights reserved.
Keyword ：
cell penetrating peptide cell penetrating peptide neutron diffraction neutron diffraction phospholipid phospholipid TAT peptide TAT peptide
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| GB/T 7714 | Chen, Xiaochao , Liu, Shutao , Deme, Bruno et al. Efficient internalization of TAT peptide in zwitterionic DOPC phospholipid membrane revealed by neutron diffraction [J]. | BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES , 2017 , 1859 (5) : 910-916 . |
| MLA | Chen, Xiaochao et al. "Efficient internalization of TAT peptide in zwitterionic DOPC phospholipid membrane revealed by neutron diffraction" . | BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES 1859 . 5 (2017) : 910-916 . |
| APA | Chen, Xiaochao , Liu, Shutao , Deme, Bruno , Cristiglio, Viviana , Marquardt, Drew , Weller, Richard et al. Efficient internalization of TAT peptide in zwitterionic DOPC phospholipid membrane revealed by neutron diffraction . | BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES , 2017 , 1859 (5) , 910-916 . |
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胶原蛋白的抗氧化活性尚未见报道,而其水解产物胶原蛋白肽的抗氧化活性报道较多.为研究造成该差异性的原因,分别采用T-AOC法、盐酸羟胺法、DPPH法和ABTS法测定胶原蛋白模拟肽、胶原蛋白肽混合物及其组成氨基酸脯氨酸和甘氨酸的抗氧化活性,并用圆二色光谱法测定CMP和CP的二级结构.试验结果表明:(1)T-AOC法中,CMP,CP和各种氨基酸都不具有抗氧化活性;(2)在盐酸羟胺法中,只有CP具有抗氧化活性;(3)在DPPH法和ABTS法中,肽和氨基酸都具有一定的抗氧化活性,且抗氧化顺序为CP＞Hypro＞CMP≥Pro＞Gly;(4)圆二色光谱法结果表明,CMP具有胶原蛋白形成稳定的3股螺旋结构所必须的α-螺旋,而CP的α-螺旋结构不明显.这些结果提示:(1)胶原蛋白本身稳定的3股螺旋结构可能是其抗氧化活性不明显的原因,而胶原蛋白肽的抗氧化活性的发挥有赖于肽键对氨基酸残基的连接作用以及氨基酸侧链对自由基的易接近性;(2)不同抗氧化测定方法对胶原蛋白肽相关分子的表征效果不同,其中DPPH法和ABTS法的灵敏度相对较高.
Keyword ：
3股螺旋结构 3股螺旋结构 抗氧化 抗氧化 胶原蛋白 胶原蛋白 胶原蛋白肽 胶原蛋白肽
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| GB/T 7714 | 林春通 , 柯欣欣 , 郭建辉 et al. 胶原蛋白与胶原蛋白肽的抗氧化关系初探 [J]. | 中国食品学报 , 2017 , 17 (10) : 44-50 . |
| MLA | 林春通 et al. "胶原蛋白与胶原蛋白肽的抗氧化关系初探" . | 中国食品学报 17 . 10 (2017) : 44-50 . |
| APA | 林春通 , 柯欣欣 , 郭建辉 , 巫启明 , 刘树滔 , 郭静科 et al. 胶原蛋白与胶原蛋白肽的抗氧化关系初探 . | 中国食品学报 , 2017 , 17 (10) , 44-50 . |
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The purpose of this study was to evaluate whether topical application of superoxide dismutase with cell penetrating peptide (HIV-TAT) could protect against skin damage induced by UVB irradiation in humans. The permeability through stratum corneum of large proteins linked to TAT peptide was firstly confirmed by confocal microscopy and tape stripping. Ten healthy volunteers with either Fitzpatrick skin type II or III were recruited in this clinical study. TAT-SOD (300 units/cm(2)) and vehicle cream were applied on two symmetric areas of both inner upper arms 1 h prior to UVB irradiation. After one hour of pretreatment, subjects received 10 incremental doses of UVB on pretreated areas. 24 h later, erythema, blood flow and apoptotic cells were measured. Pretreatment with TAT-SOD 1 h prior to UVB radiation promoted a mean minimal erythema dose (MED) increase of 36.6 +/- 18.4% (p = 0.013 < 0.05. n = 10) compared to vehicle control. The median blood flow values of all subjects following 2 and 3-MED of UVB were 107.8 +/- 51.0 units and 239.5 +/- 88.0 units respectively, which account for 26% and 25% decrease with respect to vehicle groups. These data suggest that TAT-SOD significantly suppresses UVB induced erythema formation and blood flow rise. Furthermore, pretreatment with TAT-SOD 1 h prior to 2-MED of UVB irradiation reduced the apoptotic sunburn cell formation by 47.6 +/- 8.6% (p < 0.0001) in all subjects. Evaluating results generated from all measurements, we conclude that topical application of TAT-SOD significantly attenuates UVB-induced skin damage in man. These biological effects of TAT-SOD are probably mediated via its free radical scavenging properties, clearly differentiating it from other physical sunscreen agents. (C) 2016 Elsevier B.V. All rights reserved.
Keyword ：
Blood flow Blood flow Erythema Erythema HIV-TAT peptide HIV-TAT peptide Sunburn cell Sunburn cell Superoxide dismutase Superoxide dismutase Tape stripping Tape stripping Transdermal delivery Transdermal delivery Ultraviolet Ultraviolet
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| GB/T 7714 | Chen, Xiaochao , Liu, Shutao , Rao, Pingfan et al. Topical application of superoxide dismutase mediated by HIV-TAT peptide attenuates UVB-induced damages in human skin [J]. | EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS , 2016 , 107 : 286-294 . |
| MLA | Chen, Xiaochao et al. "Topical application of superoxide dismutase mediated by HIV-TAT peptide attenuates UVB-induced damages in human skin" . | EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS 107 (2016) : 286-294 . |
| APA | Chen, Xiaochao , Liu, Shutao , Rao, Pingfan , Bradshaw, Jeremy , Weller, Richard . Topical application of superoxide dismutase mediated by HIV-TAT peptide attenuates UVB-induced damages in human skin . | EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS , 2016 , 107 , 286-294 . |
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