SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2024
·
IF 4.3
Jiangyue Ning,Xingyan Bao,Haotian Chen,Zelong Yan,Li Ding,Chang Shu
Thrombin plays a critical role in hemostasis and hemolysis, and is a significant biomarker for blood-related diseases. Detection and inhibitors screening of thrombin are essential in medical research. In this study, we developed a fluorescent sensor based on the interaction between quantum dots (QDs) and fibrinogen (Fib) for thrombin detection and its inhibitors screening. Upon the presence of thrombin, the fibrinogen of soluble QDs-Fib were converted into insoluble fibrin precipitate, causing a change of fluorescence intensity in the supernatant. Under optimized conditions, our method exhibited an excellent linearity (R 2 ≥0.99) over the range of 2∼100 U/L with a limit of detection (LOD) as low as 0.29 U/L. Moreover, we employed this method to screen for thrombin inhibitors using dabigatran as an exemplary direct thrombin inhibitor (DTI), even at concentrations as low as 1 nM. Finally, the established method was successfully used to screen thrombin inhibitors in 23 different extracts from Eupolyphaga sinensis walker . The method provided not only a sensitive, specific and high throughput assay for the detection of thrombin activity in biological samples, but also a reliable strategy for the screening of thrombin inhibitors in complex matrices.
研究领域:
生物医学工程
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分析化学
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药学
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医学研究
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
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2022
·
IF 5.999
Nan Wang,Shuying Zhao,Xiaoyong Tian,Shanyi Guang,Hongyao Xu
Traditional dressings, such as bandages, gauze, and cotton pads stick to new granulation tissue, thereby aggravating wound injury or causing secondary injury during replacement. Microspheres that are biodegradable and adaptable to various wound shapes are a good alternative to traditional dressings. In this work, a novel microsphere was prepared by reverse microemulsion method using sodium alginate and silk peptide (SP) as the aqueous phase. After cross-linking by Ca 2+ , calcium alginate (CA) and SP composite microspheres called CA/SP were prepared. By adjusting the SP content, the swelling rate of microspheres reached 1050 % and the pore diameter reached 19.59 nm. In addition, the introducing SP provided a stable loading site for thrombin (Th). This platform (called CA/SP@Th)—integrating rapid blood enrichment, calcium release , and Th catalysis—can ensure rapid hemostasis in a variety of bleeding models. Additionally, the SP present in this modality also promoted fibroblast proliferation, this increased the wound closure rate in a total cortex injury mouse model (more than 97 % within 15 days). Therefore, CA/SP@Th can be used as a multifunctional dressing for rapid hemostasis and wound healing.
研究领域:
生物材料
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医学工程
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创伤修复
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DOI
PubMed
高影响因子
ELECTROCHIMICA ACTA
·
2013
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IF 3.777
Chong Sun,Xiaobo Wang,Xujie Yang,Lei Xing,Bo Zhao,Xiaodi Yang,Chun Mao
In this paper, we reported a novel label-free electrochemical aptasensors for thrombin detection in whole blood using self-assembled multilayers with carboxymethyl-PEG-carboxymethyl (CM-PEG-CM) and thrombin-binding aptamer (TBA). In the sensing strategy, CM-PEG-CM and TBA were assembled on the electrode surface via covalent binding. In the presence of target, the TBA on the outermost layer of the self-assembled multilayer would catch the target on the electrode interface, which makes a barrier for electrons and inhibits the electro-transfer, resulting in the decreased DPV signals. Using this strategy, a wide detection range (1 pM–160 nM) for target thrombin was obtained, with a low detection limit of 1.56 × 10 −14 M. The control experiments were also carried out by using bull serum albumin (BSA) and lysozyme in the absence of thrombin. The results showed that the aptasensors had good specificity, stability and reproducibility to thrombin. Moreover, the aptasensors could be used for detection of thrombin in whole blood which could provide a promising platform for fabrication of aptamer based biosensors in clinical application.
研究领域:
生物医学工程
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分析化学
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分子诊断
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产品关联
DOI
高影响因子
Scientific Reports
·
2024
·Vol. 14
·期 1
·1-14
·
IF 3.8
Ye Jian,Shi Chenyang,Lan Jian,Chen Qingqing,Si Qin,Xu Panpan,Zhang Xijiang,Zheng Cheng
Rapid and safe hemostasis is crucial for the survival of bleeding patients in prehospital care. It is urgent to develop high performance hemostatic material to control the massive hemorrhage in the military field and accidental trauma. In this work, an efficient protein hemostat of thrombin was immobilized onto commercial gauze, which was mediated by self-polymerization and anchoring of tannic acid (TA). Through TA treatment, the efficient immobilization of thrombin was achieved, preserving both the biological activity of thrombin and the physical properties of the dressing, including absorbency, breathability, and mechanical performance. Moreover, in the presence of TA coating and thrombin, Gau@TA/Thr could obviously shortened clotting time and enriched blood components such as plasma proteins, platelets, and red blood cells, thereby exhibiting an enhanced in vitro coagulation effect. In SD rat liver volume defect and artery transection hemorrhage models, Gau@TA/Thr still had outstanding hemostatic performance. Besides, the Gau@TA/Thr gauze had inherent antibacterial property and demonstrated excellent biocompatibility. All results suggested that Gau@TA/Thr would be a potential candidate for treating uncontrollable hemorrhage in prehospital care.
研究领域:
生物医学工程
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创伤护理
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材料科学
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产品关联
DOI
PubMed
高影响因子
Chinese Medicine
·
2020
·Vol. 15
·期 1
·1-14
·
IF 2.96
Yang Yi-Yao,Wu Zhao-Yu,Xia Fang-Bo,Zhang Hao,Wang Xu,Gao Jian-Li,Yang Feng-Qing,Wan Jian-Bo
Background The dry root and rhizome of Ligusticum chuanxiong Hort., or Chuanxiong, has been used as a blood-activating and stasis-removing traditional Chinese medicine for 1000 years. Our previous studies have shown the inhibitory activity on platelet and thrombin (THR) of Chuanxiong. THR and factor Xa (FXa) play significant roles in the coagulation cascade and their inhibitors are of valuable in the treatment of thromboembolic diseases. The aim of the present study is to screen THR and FXa inhibitors from Chuanxiong.Methods Four extracts [ethyl acetate (EA), butanol (BA) and remained extract (RE) from 75% ethanol extract, and water extract (WE)] of Chuanxiong were prepared, and their THR/FXa inhibitory activities were assessed in vitro. Following silica-gel column chromatography (SC), the active EA extract and BA extract was further partitioned, respectively. Their active fractions (EA-SC1 to EA-SC5; BA-SC1 to BA-SC5) were obtained and analyzed by LC–MS. After modeling by the principal component analysis (PCA) and orthogonal partial least squares discriminate analysis (OPLS-DA), the specific marker compounds were predicted and identified. Their enzyme inhibitory was assessed in vitro and interactions with THR/FXa were investigated by molecular docking analysis.Results Chuanxiong EA extract showed strong activity against THR and BA extract was more effective in inhibiting FXa activity, and their fractions exhibited obvious difference in enzyme inhibitory activity. Furthermore, marker compounds a–h were predicted by PCA and OPLS-DA, and their chemical structures were identified. Among them, senkyunolide A, Z-ligustilide, ferulic acid and senkyunolide I (IC50 was determined as 0.77 mM) with potential THR inhibitory activity, as well as isochlorogenic acid A with FXa inhibitory activity were screened out. It was found that the four components could interact with the active site of THR, and the binding energy was lower than − 5 kcal/mol. Isochlorogenic acid A were bound to the active site of FXa, and the binding energy was − 9.39 kcal/mol. The IC50 was determined as 0.56 mM.Conclusion sTHR/FXa inhibitory components in different extracts of Chuanxiong were successfully characterized by the method of enzyme inhibition activity assays with ultra performance liquid chromatography-quadrupole time of flight mass spectrometry-based multivariate statistical analysis.
研究领域:
中药学
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生物化学
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药理学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Bioscience-Landmark
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2024
·Vol. 29
·期 9
·325
·
IF 3.3
Ziwei Guo,Sixu Bao,Zehui Shi,Xuejiao Li,Peijin Li,Bin Zhong,Ming Zhang,Qiyong Wu
Background: Platelets have the hemostatic function, and their aberrant activation is associated with occlusive thrombus formation. Plasma exosomes are rich in platelets containing ubiquitin-specific peptidase 15 (USP15). Herein, we aim to explore the effect of USP15 on thrombosis, as well as expounding whether USP15 acts as an upstream target of FK506 binding protein 5 (FKBP5) to regulate occlusive thrombus formation. Methods: Washed human platelets were treated with thrombin for measurement of USP15 and FKBP5 expressions. USP15 loss/gain-of-function variant in HEK293 cells was performed by cell transfection, and the interaction between USP15 and FKBP5 was examined using immunoprecipitation and ubiquitination assays. Mice with USP15-knockout platelets (Plt USP15-/-) were modeled, and subjected to calculation of bleeding time, artery thrombosis imaging and clot retraction assay. FKBP5 expression and the inhibitor of nuclear factor kappa B kinase subunit epsilon (IKBKE)/phosphatidylinositol 3-kinase (PI3K)/Rap1 pathway in wild-type and Plt USP15-/- mice-derived platelets were detected using Western blot. The activation of αIIbβ3 in washed platelets was analyzed using flow cytometry. Results: USP15 and FKBP5 expressions were upregulated in platelets after thrombin treatment. Following transfection of USP15 knockdown and USP15 overexpression plasmids into HEK293 cells, FKBP5 protein expression was downregulated by USP15 knockdown while being upregulated by USP15 overexpression. USP15 bound to FKBP5 and protected FKBP5 against ubiquitination. Knockdown of platelet USP15 prolonged bleeding time, inhibited arterial thrombosis and delayed clot retraction in mice. Knockdown of platelet USP15 also decreased protein expressions of FKBP5, IKBKE and Rap1, p-PI3K/PI3K ratio, and activation of αIIbβ3 in mice. Conclusion: USP15 knockdown in platelets affects thrombosis in mice by promoting the instability of FKBP5 to repress the activation of IKBKE/PI3K/Rap1 pathway-mediated αIIbβ3.
研究领域:
血液学
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分子生物学
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血栓
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细胞信号传导
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产品关联
DOI
PubMed
高影响因子
iScience
·
2021
·
IF 5.458
Dao Lin,Yuhe Kan,Liang Yan,Yongqi Ke,Yang Zhang,Hang Luo,Xinjing Tang,Xiangjun Li,Yujian He,Li Wu
Summary The effective utility of physiologically active molecules is crucial in numerous biological processes. However, the regulation of enzyme functions through active substances remains challenging at present. Here, glutathione (GSH), produced in cells, was used to modulate the catalytic activity of thrombin without external stimulus. It was found that high concentrations of GSH was more conducive to initiate the cleavage of compound AzoDiTAB in the range of concentration used to mimic the difference between cancer and normal cells, which has practical implications for targeting cancel cells since GSH is overexpressed in cancer cells. Importantly, GSH treatment caused the deformation of G4 structure by cleaving AzoDiTAB and thus triggered the transition of thrombin from being free to be inhibited in complex biological systems. This work would open up a new route for the specific manipulation of enzyme-catalyzed systems in cancer cells.
研究领域:
生物化学
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分子生物学
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癌症研究
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2015
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IF 2.858
Jing-jing Rong,Ming Liang,Feng-qi Xuan,Jing-yang Sun,Li-jun Zhao,Hui-zhen Zhen,Xiao-xiang Tian,Dan Liu,Quan-yu Zhang,Cheng-fei Peng,Tian-ming Yao,Fei Li,Xiao-zeng Wang,Ya-ling Han,Wei-ting Yu
To date, transcatheter arterial embolization (TAE) has become a standard treatment to control intracavitary bleeding as an alternative to surgery. Due to excellent biocompatibility and no residual in vivo , biodegradable materials are preferred in TAE. However, gelfoam is the only commercially available biodegradable embolic material used to treat blunt trauma of solid abdominal viscera until now, and controversial on its stability and reliability never stopped in the past five decades. In this study, a new biodegradable macromolecule material (thrombin-loaded alginate-calcium microspheres, TACMs) was prepared using electrostatic droplet techniques and a special method was developed for hemostatic embolization. Thrombin was successfully loaded into microspheres with high encapsulation efficiency and drug loading capacity. A burst release of TACMs was observed at early stage and sustained release later on, with the activity of thrombin preserved well. The strength of TACMs mixed thrombus, which was used as embolic agent, increased in a dose-dependent manner after TACMs were added. In addition, the TACMs were verified to be of no cytotoxicity and systemic toxicity, and biodegradable in vivo . Finally, the results of preliminary applications revealed that the TACMs could serve as an effective and promising embolic material for blunt trauma and hemorrhage of solid abdominal viscera.
研究领域:
医学
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生物材料学
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产品关联
DOI
PubMed
高影响因子
SENSORS
·
2016
·Vol. 16
·期 1
·135
·
IF 2.033
Jiexia Chen,Yifan Liu,Guang-Chao Zhao
A novel photoelectrochemical biosensor for step-by-step assay of tyrosinase and thrombin was fabricated based on the specific interactions between the designed peptide and the target enzymes. A peptide chain with a special sequence which contains a positively charged lysine-labeled terminal, tyrosine at the other end and a cleavage site recognized by thrombin between them was designed. The designed peptide can be fixed on surface of the CdTe quantum dots (QDs)-modified indium-tin oxide (ITO) electrode through electrostatic attraction to construct the photoelectrochemical biosensor. The tyrosinase target can catalyze the oxidization of tyrosine by oxygen into ortho-benzoquinone residues, which results in a decrease in the sensor photocurrent. Subsequently, the cleavage site could be recognized and cut off by another thrombin target, restoring the sensor photocurrent. The decrease or increase of photocurrent in the sensor enables us to assay tyrosinase and thrombin. Thus, the detection of tyrosinase and thrombin can be achieved in the linear range from 2.6 to 32 μg/mL and from 4.5 to 100 μg/mL with detection limits of 1.5 μg/mL and 1.9 μg/mL, respectively. Most importantly, this strategy shall allow us to detect different classes of enzymes simultaneously by designing various enzyme-specific peptide substrates.Keywords:tyrosinase;thrombin;peptide;photoelectrochemical biosensor
研究领域:
生物化学
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医学检测
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纳米技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Xiujun Cao,Yuan Luo,Xueli Liu,Chunqing Shang,Jun Lu,Guoxin Song,Chunhui Deng
Hirudin, that is naturally occurring in leeches ( Hirudo medicinalis ) and known as the most potent natural inhibitor of thrombin, exerts double-edged effects in clinic application. It can be used as a therapeutic ingredient for cardiovascular disease, while it can be regarded as a toxic polypeptide with bleeding risk. Effective detection of hirudin in biological samples contributes greatly to reasonable therapy. In this study, we proposed a smart adsorbent based on affinity magnetic microspheres , where thrombin was immobilized for capturing hirudin in the animal serum. Aptamer was introduced as a ligand for linking the magnetic agarose microspheres and thrombin, thereby avoiding loss of biological activity of the enzyme to hirudin. Taken recombinant hirudin variant 2-Lys 47 (rHV2) as a model, we established a rapid and bio-specific extraction method coupled with liquid chromatography and quadrupole-time-of-flight mass spectrometry (LC-QTOF/MS) for determination of hirudin in the serum. Owing to this strategy, a low limit of detection (LOD) of rHV2 (0.5 nM), a good linearity with correlation coefficient of 0.9975, an acceptable precision with relative standard deviation (RSD) below 3.6% (n = 6) and acceptable recoveries ranging from 85.7% to 90.2% were achieved. Moreover, the functionalized magnetic composite could be reused for at least nine cycles. Our work combined the merits of affinity separation and advanced instrument analysis for hirudin, providing a new vision to precise determination of hirudin in medical and pharmaceutical fields.
研究领域:
生物化学
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药物分析
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医学检测
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Huijuan Zhang,Kexuan Cao,Chaoqun Wang,Yingmei Tian,Lin Hou
Drug thrombolysis is the main means of clinical treatment for thrombotic-related diseases. However, serious bleeding complications caused by low drug delivery efficiency and secondary vascular embolism usually lead to unsatisfactory therapeutic effects. Herein, we constructed a thrombin and near-infrared (NIR) dual-responsive drug-loaded intelligent system to solve the above problems. The CREKA-modified polydopamine (PDA) nanoparticles (NPs) were crosslinked by thrombin-responsive peptide (Pep), to obtain Pep-PPC nanocarriers with special mesoporous nanostructures. Subsequently, NO donor BNN6 and thrombolytic drug UK were loaded on the surface and into the mesoporous of Pep-PPC, to get the multifunctional Pep-NO@PPC/UK. In vitro and in vivo results proved that Pep-NO@PPC/UK could effectively recognize and aggregate at the thrombus site via CREKA-mediated active targeting. Then local abundant thrombin cleaved the nanoplatform to release UK. Meanwhile, PDA converted NIR light into heat and synchronously triggered NO release. This transformed the DDS into a “heat-gas” dual propulsion nanomotor in situ, driving deep penetration of UK in thrombus tissue. Under the combined action of UK and PTT, arterial thrombosis rapidly dissolved with relatively low bleeding risk. More importantly, NO generated in situ could prevent re-embolism of blood vessels at the thrombolytic site by inhibiting activated platelet aggregation, ultimately improving vascular reperfusion rate through a combination of attack and defense mode.
研究领域:
药学
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纳米技术
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生物医学工程
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治疗学
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产品关联
DOI
高影响因子
NPG Asia Materials
·
2022
·Vol. 14
·期 1
·1-12
·
IF 10.761
Shen Peiyan,Li Mengru,Li Ran,Han Bo,Ma Haojie,Hou Xueyan,Zhang Yuqi,Wang Ji-Jiang
Smart photonic hydrogels based on two-dimensional photonic crystals (2DPC) provide a promising sensing platform for constructing novel chemical and biological sensors due to their facile optical signal readout and highly sensitive responsivity toward target analytes. Aptamers, as recognition elements with high selectivity and affinity, are extensively used to construct a variety of sensors. Herein, we developed two partially base complementary aptamer-functionalized 2DPC hydrogels as aptasensors for the detection of thrombin (TB) in human serum. The photonic hydrogel aptasensors swelled upon exposure to TB solution, leading to an increase in the particle spacing of the 2DPCs. The particle spacing changes were acquired by simply measuring the diameters of the Debye ring diffracted by the 2DPCs without the requirement of sophisticated instruments. The aptasensor swelling resulted from the decrease in the hydrogel cross-linking density induced by the specific binding between one of the aptamers and TB and the increase in hydrogel mixing free energy induced by the introduction of TB. The particle spacing increase of the optimized aptasensor was linear over the TB concentration range of 1–500 nM, and the limit of detection was 0.64 nM. The constructed 2DPC hydrogel aptasensor was used to detect TB in human serum and achieved recoveries of 95.74–104.21% and a relative standard deviation of 2.52–6.58%, showing the practicability and accuracy of the sensor. The aptamer-actuated 2DPC hydrogel biosensor provides a new strategy for designing other target molecule-sensitive aptasensors, showing great potential for development into home kits.
研究领域:
生物传感
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材料科学
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医学检测
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DOI
高影响因子
危险品化学品经营许可证（带存储）