ANALYTICAL CHEMISTRY
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2024
·
IF 6.7
Liming Liu,Ning Ma,Lu Wang,Yu Zhang,Yi-Zhen Wan,Tianze Wang,Weiping Qian
Fibrinolytic activity assay is particularly important for the detection, diagnosis, and treatment of cardiovascular disease and the development of fibrinolytic drugs. A novel efficacious strategy for real-time and label-free dynamic detection of fibrinolytic activity based on ordered porous layer interferometry (OPLI) was developed. Fibrin or a mixture of fibrin and plasminogen (Plg) was loaded into the highly ordered silica colloidal crystal (SCC) film scaffold to construct a fibrinolytic response interference layer to measure fibrinolytic activity with different mechanisms of action. Fibrinolytic enzyme-triggered fibrinolysis led to the migration of interference fringes in the interferogram, which could be represented by optical thickness changes (ΔOT) tracked in real time by the OPLI system. The morphology and optical property of the fibrinolytic response interference layer were characterized, and the Plg content in the fibrinolytic response interference layer and experimental parameters of the system were optimized. The method showed adequate sensitivity for the fibrinolytic activity of lumbrokinase and streptokinase, with wide linear ranges of 12–6000 and 10–2000 U/mL, respectively. Compared with the traditional fibrin plate method, it has a lower detection limit and higher linearity. The whole kinetic process of fibrinolysis by these two fibrinolytic drug models was recorded in real time, and the Michaelis constant and apparent kinetic parameters were calculated. Importantly, some other blood proteins were less interfering with this system, and it showed reliability in fibrin activity detection in real whole blood samples. This study established a better and more targeted research method of in vitro fibrinolysis and provided dynamic monitoring data for the analysis of fibrinolytic activity of whole blood.
研究领域:
生物医学工程
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心血管疾病
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药物开发
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干涉测量
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纤溶
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Shengjie Liu,He Yang,Xingyu Heng,Lihua Yao,Wei Sun,Qing Zheng,Zhaoqiang Wu,Hong Chen
To effectively solve the problem of significant loss of transplanted cells caused by thrombosis during cell transplantation, this study simulates the human fibrinolytic system and combines metabolic oligosaccharide engineering with strain-promoted azide–alkyne cycloaddition (SPAAC) click chemistry to construct a cell surface with fibrinolytic activity. First, a copolymer (POL) of oligoethylene glycol methacrylate (OEGMA) and 6-amino-2-(2-methylamido)hexanoic acid (Lys) was synthesized by reversible addition–fragmentation chain transfer (RAFT) copolymerization, and the dibenzocyclooctyne (DBCO) functional group was introduced into the side chain of the copolymer through an active ester reaction, resulting in a functionalized copolymer DBCO-PEG4-POL with ε-lysine ligands. Then, azide functional groups were introduced onto the surface of HeLa model cells through metabolic oligosaccharide engineering, and DBCO-PEG4-POL was further specifically modified onto the surface of HeLa cells via the SPAAC “click” reaction. In vitro investigations revealed that compared with unmodified HeLa cells, modified cells not only resist the adsorption of nonspecific proteins such as fibrinogen and human serum albumin but also selectively bind to plasminogen in plasma while maintaining good cell viability and proliferative activity. More importantly, upon the activation of adsorbed plasminogen into plasmin, the modified cells exhibited remarkable fibrinolytic activity and were capable of promptly dissolving the primary thrombus formed on their surfaces. This research not only provides a novel approach for constructing transplantable cells with fibrinolytic activity but also offers a new perspective for effectively addressing the significant loss of transplanted cells caused by thrombosis.
研究领域:
生物化学
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细胞生物学
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材料科学
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药学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
Foods
·
2024
·Vol. 14
·期 1
·3
·
IF 4.7
Nan Li,Zhigang Yang,Zhiyu Yuan,Zizhu Zhen,Xinna Xie,Danqing Zhu,Gang Lu,Feng Zhao,Bo Qu,Bingli Qi,Yujun Jiang,Qianyu Zhao,Chaoxin Man
This study investigated the variations and alterations in the concentrations of plasmin system components in raw and UHT (ultra-high-temperature) milk under cold stress (WCT ≤ −25 °C), heat stress (THI ≥ 80), and normal (THI < 70 and WCT ≥ −10 °C) circumstances. The findings indicated elevated amounts of plasmin system components in cold-stressed raw milk. While storing UHT milk at 25 °C, the concentrations and activity of plasmin in the milk exhibited an initial increase followed by a decrease, peaking around the 30th day. The maximum plasmin level and activity in cold-stressed milk were 607.86 μg/L and 15.99 U/L, respectively, with the beginning of gelation occurring around day 60. The higher activity of plasmin in cold-stressed milk led to the poorer stability and sensory assessment of the milk. However, heat-stressed milk is not such a problem for UHT milk as cold-stressed milk. The findings indicate shortcomings in the quality of cold-stressed milk and its adverse effects on the stability of UHT milk, underscoring the necessity of preventing cold stress in the herd and refraining from utilizing cold-stressed milk as a raw material for UHT production.
研究领域:
食品科学
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生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Wenli Zhang,Maoyuan Sun,Yun Liu,Yu Zhang,Lian Xu,Ying Luo,Qianying Du,Jie Xu,Jia Liu,Jun Zhou,Haitao Ran,Zhigang Wang,Junrui Wang,Dajing Guo
Thrombotic diseases cause significant risks of disability and death worldwide. However, conventional thrombolytic drugs have short half-lives, poor targeting capabilities, limited therapeutic effects， and narrow therapeutic windows. Moreover, most basic studies focus only on thrombolytic efficiency and ignore the inflammatory factors that promote thrombosis progression, such as reactive oxygen species (ROS)-mediated oxidative stress. Therefore, to achieve comprehensive and effective thrombolytic management, both targeted thrombolytic therapy and the elimination of inflammatory factors must be achieved. In this study, we developed a platelet membrane (PM)-functionalized hollow mesoporous Prussian blue nanomedicine (HMPB-rtPA@PM), which was innovatively coassembled from two FDA-approved drugs (Prussian blue (PB) and alteplase (rtPA)) and showed marked advantages in both thrombi targeting and photothermal/pharmacological synergistic fibrinolysis. Moreover, HMPB-rtPA@PM exhibited the potential to inhibit platelet aggregation and regulate the inflammatory microenvironment by scavenging ROS. As expected, this nanomedicine not only achieved more efficient thrombolysis in both thrombus models but also delayed thrombosis progression through therapeutic visualization in the black tail model. Such rational therapeutic principle provides a broad platform for the comprehensive management of thrombotic diseases.
研究领域:
药学
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纳米医学
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血栓研究
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药物递送系统
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产品关联
DOI
高影响因子
ACS Applied Bio Materials
·
2024
·
IF 4.6
Xiaolan Mai,Yu Hu,Zhenlin Wu,Xin Guo,Mingming Dong,Lingyun Jia,Jun Ren
Recent clinical studies have highlighted the presence of microclots in the form of amyloid fibrinogen particles (AFPs) in plasma samples from Long COVID patients. However, the clinical significance of these abnormal, nonfibrillar self-assembly aggregates of human fibrinogen remains debated due to the limited understanding of their structural and biological characteristics. In this study, we present a method for generating mimetic microclots in vitro. Using this approach, the self-assembly process, structural organization of AFPs, and their interactions with human plasma components were elucidated. The amyloid transition of fibrinogen occurs under acidic conditions within a pH range of 2.3–3.2. Well-dispersed amyloid oligomers of fibrinogen, ranging in size from 1 to 5 μm, can be prepared at pH 2.8 after 1 h of incubation. We tracked the dynamic self-assembly process at the single-molecule level using high-speed atomic force microscopy (HS-AFM). The arrangement of amyloid oligomers manifests as well-ordered, stacked nanodomains with striped patterns, growing perpendicular to the primary axis of the fibrinogen monomer. Upon transfer to physiological solution conditions or human plasma, these amyloid oligomers further aggregate into nonfibrillar structures at the micrometer scale, resembling the microclots observed in the bloodstream of Long COVID patients. Notably, these AFPs exhibit characteristics consistent with microclots, including positive staining in thioflavin T (ThT) assays and resistance to fibrinolysis. Proteomic analysis suggests that AFPs interact with various components of human plasma and have an enhanced binding affinity with complement C3 compared to native fibrinogen. This study enables the in vitro preparation of mimetic microclots exhibiting amyloid features. It is anticipated to facilitate further researches on the mechanisms, detection, and treatment of diseases associated with fibrinogen amyloidogenesis.
研究领域:
生物化学
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医学研究
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产品关联
DOI
PubMed
高影响因子
Polymer Chemistry
·
2013
·Vol. 4
·期 22
·5597-5602
·
IF 5.231
Zengchao Tang,Xiaoli Liu,Yafei Luan,Wei Liu,Zhaoqiang Wu,Dan Li,Hong Chen
A new strategy to regulate the attachment of plasminogen and its activator, t-PA, to a polyurethane surface is reported. The method is based on the one-step graft copolymerization of a lysine-containing methacrylic monomer and hydroxyethyl methacrylate (HEMA). The copolymer-modified PU surfaces showed high capacity for plasminogen and t-PA binding. The release of pre-adsorbed t-PA from the lysine-containing PU surfaces in contact with plasma was also investigated. It was shown that the interactions of plasminogen and t-PA on this type of surface can be controlled simply by changing the monomer feed ratio, and thus the relative quantities of HEMA and lysine in the grafts. This approach may be useful for the development of fibrinolytic, blood-contacting materials and, more generally, for controlling protein-surface interactions for biocompatibility.
研究领域:
生物材料学
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医学工程
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DOI
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2023
·
IF 3.1
Chunying Guo,Ting Wang,Dongmei Zhang,Xiaojing Ge,Jinan Li
Alzheimer's disease (AD) is the most common neurodegenerative disorder in the world. The aggregation of both amyloid beta (Aβ) peptides extracellularly and Tau proteins intracellularly plays key roles in the pathological consequences of AD, which lead to cholinergic neurodegeneration and eventually death. Currently, there are no effective methods to stop the progression of AD. Using ex vivo , in vivo and clinical approaches, we investigated the functional effects of plasminogen on the widely used FAD, Aβ42 oligomer or Tau intracranial injection-induced AD mouse model and explored its therapeutic effects on patients with AD. The results show that intravenously injected plasminogen rapidly crosses the blood‒brain barrier (BBB); increases plasmin activity in the brain; colocalizes with and effectively promotes the clearance of Aβ42 peptide and Tau protein deposits ex vivo and in vivo ; increases the choline acetyltransferase (ChAT) level and decreases the acetylcholinesterase (AChE) activity; and improves the memory functions. Clinically, when GMP-level plasminogen was administered to 6 AD patients for 1–2 weeks, their average scores on the Minimum Mental State Examination (MMSE), which is a standard scoring system used to measure the memory loss and cognitive deficits, were extremely significantly improved by 4.2 ± 2.23 points, e.g., an average increase from 15.5 ± 8.22 before treatment to 19.7 ± 7.09 after treatment. The preclinical study and pilot clinical study suggest that plasminogen is effective in treating AD and may be a promising drug candidate.
研究领域:
神经科学
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药理学
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临床试验
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产品关联
DOI
PubMed
高影响因子
Colloid and Interface Science Communications
·
2025
·
IF 4.7
Jun Sun,Sulei Zhang,Yichen Wang,Denghai Sheng,Shengjie Liu,Yu Rao,Aiqing Li,Yuchun Pan,John L. Brash,Xiaoli Liu,Hong Chen
Thrombus formation often leads to the failure of intravascular implants. Natural endothelium provides multifaceted antithrombotic functions through nitric oxide/ prostacyclin secretion to inhibit platelet activation, glycosaminoglycan mediated anticoagulation, and tissue-type plasminogen activator driven fibrinolysis. Therefore, surfaces mimicking these multiple endothelial functions are expected to have enhanced antithrombotic properties. In this study, polyvinyl chloride surface was rendered porous through solvent/nonsolvent-induced phase separation and loaded with a metal-organic framework, CuBTTri to catalyze nitric oxide release from a precursor. Furthermore, using layer-by-layer self-assembly, multiple bilayers of a poly(lysine- co -oligo(ethylene glycol) methyl ether methacrylate) copolymer (fibrinolysis-promoting), and sodium heparin (endothelial cell growth-promoting), were deposited on the un-etched side of the polyvinyl chloride. This modified surface was shown to be capable of releasing nitric oxide, destroying nascent thrombus, inhibiting smooth muscle cell growth, and promoting endothelial cell adhesion. This study represents a novel approach to developing multifunctional blood-contacting surfaces that mimic multiple properties of the endothelium.
研究领域:
生物医学工程
·
材料科学
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产品关联
DOI
高影响因子
Food Chemistry-X
·
2024
·
IF 6.5
Wentao Qian,Xiaobing Wang,Hongliang Li,Yinhua Zhu,Pengjie Wang,Xiaolu Geng,Jinhui Yang,Huiyuan Guo,Menghui Wang,Chong Chen
Introducing Holstein cows on Qinghai-Tibetan Plateau is a potential solution to enhance local milk production. However, the relationship between milk quality and altitude in China remains unknown. Therefore, the components and plasmin (PL) system of raw milk from different altitudes (sea level, 1600, 2700, and 3800 m) were investigated. The daily milk production of Holstein cows and PL activity decreased as the altitude increased. However, the components content of raw milk, plasminogen (PLG)/PL ratio, activities of PLG and plasmin activator (PA) increased with altitude. The pasteurization resulted a significant decrease in PA activity of all milk and a significant increase in PL activity in milk collected at higher altitudes (2700 and 3800 m), suggesting the pasteurization was unsuitable for preserving milk at higher altitudes. This study offered references for the production and storage of milk after introducing Holstein cows on Qinghai-Tibetan Plateau.
研究领域:
农业
·
乳品科学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2017
·Vol. 142
·期 24
·4773-4781
·
IF 3.885
Haijiao Zheng,Naizhong Song,Xiqian Li,Qiong Jia
Plasminogen (Plg) is a kind of glycoprotein which plays an important role in cell migration. The determination of Plg content can directly reflect the abnormal manifestation of fibrinolytic system dysfunctions. In the present work, lysine (Lys)-based adsorbents were prepared for the specific capture of Plg through the covalent binding of Lys with a polymer monolithic substrate. Lys was modified with β-cyclodextrin (β-CD) via a click reaction and anchored to the substrate with biotin by host–guest interaction. The biotin-Lys-CD based monolithic material was employed for the specific capture of Plg. Combining with mass spectrometry (MS) determinations, the method exhibited a low detection limit of 1.0 fmol with relative standard deviations below 10.0% for Plg. Considering the advantages of simplicity, sensitivity, and high specificity, the developed approach was successfully applied to the determination of Plg in human plasma samples and opened a gallery for testing Plg as a biomarker for the diagnosis of fibrinolytic system dysfunctions.
研究领域:
生物化学
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
Marine Drugs
·
2024
·Vol. 22
·期 2
·68
·
IF 4.9
Tingting Jiang,Bing Zhang,Haixing Zhang,Mingjun Wei,Yue Su,Tuo Song,Shijia Ye,Yuping Zhu,Wenhui Wu
Marine organisms are a rich source of enzymes that exhibit excellent biological activity and a wide range of applications. However, there has been limited research on the proteases found in marine mudflat organisms. Based on this background, the marine fibrinolytic enzyme FELP, which was isolated and purified from clamworm (Perinereis aibuhitensis), has exhibited excellent fibrinolytic activity. We demonstrated the FELP with a purification of 10.61-fold by precipitation with ammonium sulfate, ion-exchange chromatography, and gel-filtration chromatography. SDS-PAGE, fibrin plate method, and LC–MS/MS indicated that the molecular weight of FELP is 28.9 kDa and identified FELP as a fibrinolytic enzyme-like protease. FELP displayed the maximum fibrinolytic activity at pH 9 (407 ± 16 mm2) and 50 °C (724 ± 27 mm2) and had excellent stability at pH 7–11 (50%) or 30–60 °C (60%), respectively. The three-dimensional structure of some amino acid residues of FELP was predicted with the SWISS-MODEL. The fibrinolytic and fibrinogenolytic assays showed that the enzyme possessed direct fibrinolytic activity and indirect fibrinolysis via the activation of plasminogen; it could preferentially degrade Aα-chains of fibrinogen, followed by Bβ- and γ-chains. Overall, the fibrinolytic enzyme was successfully purified fromPerinereis aibuhitensis,a marine Annelida (phylum), with favorable stability that has strong fibrinolysis activity in vitro. Therefore, FELP appears to be a potent fibrinolytic enzyme with an application that deserves further investigation.Keywords:Perinereis aibuhitensis;plasmin-like protease;amino acid residue sequences;fibrinolysis in vitro
研究领域:
海洋生物学
·
酶学
·
生物化学
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产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2015
·
IF 4.457
Xiujuan Shi,Wenjun Zhan,Gaojian Chen,Qian Yu,Qi Liu,Hui Du,Limin Cao,Xiaoli Liu,Lin Yuan,Hong Chen
The protein binding capability of biomaterial surfaces can significantly affect subsequent biological responses, and appropriate ligand presentation is often required to guarantee the best functions. Herein, a new facile method for regulating this capability by varying the localized and average ligand density is presented. Binding between lysine and plasminogen relevant to a fibrinolysis system was chosen as a model. We integrated different lysine-modified β-cyclodextrin (β-CD) derivatives onto bioinert copolymer brushes via host–guest interactions. The localized and average lysine density can be conveniently modulated by changing the lysine valency on β-CD scaffolds and by diluting lysine-persubstituted β-CD with pure β-CD, respectively. Both the plasminogen adsorption and the plasminogen binding affinity were enhanced by lysine-persubstituted β-CD compared with those of lysine-monosubstituted β-CD, which is possibly due to the higher localized lysine density and the multivalent binding of plasminogen on lysine-persubstituted β-CD surfaces. With a change in the ratio of lysine-persubstituted β-CD to β-CD, the average lysine density can be tuned, leading to the linear regulation of the adsorption of plasminogen on surfaces.
研究领域:
生物材料学
·
分子生物学
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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）