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1.
Virus Res ; 338: 199234, 2023 12.
Article in English | MEDLINE | ID: mdl-37802295

ABSTRACT

Dengue virus (DENV) is one of the most prevalent arthropod-borne diseases. It may cause dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), while no effective vaccines and drugs are available. Our study demonstrated that conessine exhibits broad antiviral activity against several enveloped viruses, including DENV, vesicular stomatitis virus, and herpes simplex virus. In addition, conessine has no direct destructive effect on the integrity or infectivity of virions. Both pre-treatment and post-treatment with conessine significantly reduce DENV replication. Pre-treatment with conessine disrupts the endocytosis of enveloped viruses, while post-treatment disturbs DENV RNA replication or translation at an early stage. Through screening differentially expressed genes by transcriptome sequencing, we found that conessine may affect cholesterol biosynthesis, metabolism or homeostasis. Finally, we confirmed that conessine inhibits virus replication through up-regulating cholesterol levels. Our work suggests that conessine could be developed as a prophylactic and therapeutic treatment for infectious diseases caused by enveloped viruses.


Subject(s)
Dengue Virus , Dengue , Humans , Dengue Virus/physiology , Cholesterol/pharmacology , Virus Replication , Antiviral Agents/pharmacology , Antiviral Agents/therapeutic use
2.
Front Endocrinol (Lausanne) ; 14: 1170526, 2023.
Article in English | MEDLINE | ID: mdl-37664836

ABSTRACT

Background: Gemcitabine (GEM) is a second-line anticancer drug of choice for some colorectal cancer (CRC) patients, and GEM inability to be commonly available in the clinic due to the lack of clarity of the exact action targets. Methods: The half maximal inhibitory concentration (IC50) of GEM treatment for 42 CRC cell lines were accessed from the Genomics of Drug sensitivity in Cancer (GDSC) database. High-throughput sequencing data of CRC patients were captured in The Cancer Genome Atlas (TCGA) and Weighted correlation network analysis (WGCNA) was conducted. Pearson correlations were derived for GEM potency-related genes. Differential analysis was conducted in the TCGA cohort to obtain CRC development-related genes (CDRGs), and univariate COX model analysis was performed on CDRGs overlapping with GEM potency-related genes to obtain CDRGs affecting CRC prognosis. Hub genes affecting GEM potency were identified by Spearman correlation. Results: CALB2 and GPX3 were identified as potential targets for GEM treatment of CRC via prognostic analysis, which we also observed to be elevated with elevated clinical stage in CRC patients. The enhanced expression of CALB2 and GPX3 genes identified in the pathway analysis might inhibit the body metabolism as well as activate immune and inflammation related pathways. In addition, we found that CALB2 and GPX3 could also be considered as prognostic biomarkers in pan-cancer. Finally, we found that CALB2 and GPX3 were remarkably associated with the drug sensitivity of MG-132, Dasatinib, Shikonin, Midostaurin, MS-275, and Z-LNle-CHO, which were expected to be the drugs of choice for GEM combination. Conclusion: CALB2 and GPX3 represent prognostic biomarkers for CRC and they might be potential action targets for GEM. Our study offered innovative ideas for GEM administration strategies.


Subject(s)
Colorectal Neoplasms , Gemcitabine , Humans , Cell Line , Dasatinib , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/genetics , Biomarkers
3.
Clinics (Sao Paulo) ; 78: 100213, 2023.
Article in English | MEDLINE | ID: mdl-37269788

ABSTRACT

OBJECTIVES: Pain is associated with many circumstances, including inflammatory reactions, which arise from modification of the features of signaling pathways. α2-adrenergic receptor antagonists are widely utilized in narcosis. Here, the authors focused on the narcotic effect of A-80426 (A8) on Complete Freund's Adjuvant (CFA) injections-triggered chronic inflammation pain in WT and TRPV1-/- mice and explored whether its antinociceptive impact was modulated via Transient Receptor Potential Vanilloid 1 (TRPV1). METHOD: CFA with or without A8 was co-administered to the mice, which were categorized randomly into four groups: CFA, A8, control, and vehicle. Pain behaviors underwent evaluation through mechanical withdrawal threshold, abdominal withdrawal reflex, and thermal withdrawal latency of WT animals. RESULTS: Quantitative polymerase chain reaction revealed that inflammation-promoting cytokines (IL-1ß, IL-6, and TNF-α) were upregulated in Dorsal Root Ganglion (DRG) and Spinal Cord Dorsal Horn (SCDH) tissues of WT animals. A8 administration reduced the pain behaviors and production of pro-inflammatory cytokines; however, this effect was significantly reduced in TRPV1-/- mice. Further analysis showed that CFA treatment reduced the TRPV1 expression in WT mice and A8 administration increased its expression and activity. The co-administration of SB-705498, a TRPV1 blocker, did not influence the pain behaviors and inflammation cytokines in CFA WT mice; however, SB-705498 the effect of A8 in WT mice. In addition, the TRPV1 block decreased the NFκB and PI3K activation in the Dorsal Root Ganglia (DRG) and Spinal Cord Dorsal Horn (SCDH) tissues of WT mice. CONCLUSIONS: Together, A8 exerted a narcotic impact on CFA-supplemented mice via the TRPV1-modulated NFκB and PI3K pathway.


Subject(s)
Antineoplastic Agents , Phosphatidylinositol 3-Kinases , Mice , Animals , Freund's Adjuvant/adverse effects , Phosphatidylinositol 3-Kinases/metabolism , TRPV Cation Channels/adverse effects , TRPV Cation Channels/metabolism , Pain/drug therapy , Cytokines , NF-kappa B/metabolism , Antineoplastic Agents/adverse effects , Inflammation
4.
Front Bioeng Biotechnol ; 11: 1207048, 2023.
Article in English | MEDLINE | ID: mdl-37362207

ABSTRACT

Introduction: Drosophila melanogaster is a model organism for studying developmental biology and human neural disorders. Nanobodies are the variable domains of the heavy chains of camelid heavy-chain antibodies (VHHs) with high affinity to their antigens and have applications in basic research, similar to traditional antibodies. In addition, nanobodies acting as functionalized antibodies or protein binders have become an additional valuable approach in Drosophila. This study aimed to develop a VHH library against Drosophila proteins and confirm its availability by retrieving some Drosophila protein-specific nanobodies from the library. Methods: An alpaca was first immunized with Drosophila embryo lysate and then its lymphocytes were isolated. Total RNA was extracted and cDNA was synthesized. The vhh sequences were amplified by two round PCR, which were then ligated to a phage display vector pADL-10b. The ligation products were transduced into SS320 competent cells to generate a VHH library. From this library, nanobodies against CG7544, Myc, and CyclinE was enriched and screened by phage display technology and ELISA. DNA sequences of identified nanobodies were cloned into pADL-10b-Flag-His for expression and purification in Escherichia coli SS320. Binding ability of purified nanobodies with corresponding antigens were determined by ELISA and surface plasmon resonance in vitro. Results: In this study, an immune VHH library against Drosophila embryo proteins was generated with a capacity of 3 × 107. From this library, eight nanobodies against three Drosophila proteins, Myc, CyclinE, and CG7544, were identified and the DNA sequences of these nanobodies were obtained. These nanobodies were successfully expressed and purified from Escherichia coli SS320, and were demonstrated to bind corresponding antigens with high affinity in vitro. Moreover, the equilibrium constant between the highest enriched nanobodies and corresponding antigens were calculated. Conclusion: In summary, we report the availability of an immune VHH library and a highly efficient panning strategy for nanobodies against proteins in Drosophila.

5.
Clinics ; 78: 100213, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1447980

ABSTRACT

Abstract Objectives Pain is associated with many circumstances, including inflammatory reactions, which arise from modification of the features of signaling pathways. α2-adrenergic receptor antagonists are widely utilized in narcosis. Here, the authors focused on the narcotic effect of A-80426 (A8) on Complete Freund's Adjuvant (CFA) injections-triggered chronic inflammation pain in WT and TRPV1-/- mice and explored whether its antinociceptive impact was modulated via Transient Receptor Potential Vanilloid 1 (TRPV1). Method CFA with or without A8 was co-administered to the mice, which were categorized randomly into four groups: CFA, A8, control, and vehicle. Pain behaviors underwent evaluation through mechanical withdrawal threshold, abdominal withdrawal reflex, and thermal withdrawal latency of WT animals. Results Quantitative polymerase chain reaction revealed that inflammation-promoting cytokines (IL-1β, IL-6, and TNF-α) were upregulated in Dorsal Root Ganglion (DRG) and Spinal Cord Dorsal Horn (SCDH) tissues of WT animals. A8 administration reduced the pain behaviors and production of pro-inflammatory cytokines; however, this effect was significantly reduced in TRPV1-/- mice. Further analysis showed that CFA treatment reduced the TRPV1 expression in WT mice and A8 administration increased its expression and activity. The co-administration of SB-705498, a TRPV1 blocker, did not influence the pain behaviors and inflammation cytokines in CFA WT mice; however, SB-705498 the effect of A8 in WT mice. In addition, the TRPV1 block decreased the NFκB and PI3K activation in the Dorsal Root Ganglia (DRG) and Spinal Cord Dorsal Horn (SCDH) tissues of WT mice. Conclusions Together, A8 exerted a narcotic impact on CFA-supplemented mice via the TRPV1-modulated NFκB and PI3K pathway.

6.
J Pharm Anal ; 12(4): 627-636, 2022 Aug.
Article in English | MEDLINE | ID: mdl-36105160

ABSTRACT

Alzheimer's disease (AD) represents the main form of dementia; however, valid diagnosis and treatment measures are lacking. The discovery of valuable biomarkers through omics technologies can help solve this problem. For this reason, metabolomic analysis using ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) was carried out on plasma, hippocampus, and cortex samples of an AD rat model. Based on the metabolomic data, we report a multi-factor combined biomarker screening strategy to rapidly and accurately identify potential biomarkers. Compared with the usual procedure, our strategy can identify fewer biomarkers with higher diagnostic specificity and sensitivity. In addition to diagnosis, the potential biomarkers identified using our strategy were also beneficial for drug evaluation. Multi-factor combined biomarker screening strategy was used to identify differential metabolites from a rat model of amyloid beta peptide 1-40 (Aß1-40) plus ibotenic acid-induced AD (compared with the controls) for the first time; lysophosphatidylcholine (LysoPC) and intermediates of sphingolipid metabolism were screened as potential biomarkers. Subsequently, the effects of donepezil and pine nut were successfully reflected by regulating the levels of the abovementioned biomarkers and metabolic profile distribution in partial least squares-discriminant analysis (PLS-DA). This novel biomarker screening strategy can be used to analyze other metabolomic data to simultaneously enable disease diagnosis and drug evaluation.

7.
Aging (Albany NY) ; 14(16): 6507-6519, 2022 08 16.
Article in English | MEDLINE | ID: mdl-35972910

ABSTRACT

Some microRNAs (miRNAs) play important roles in lung ischemia-reperfusion injury (LIRI) injury. Here, this study aimed to examine whether miR-141 was related to lung ischemia-reperfusion injury (IRI) via regulating autophagy and the epidermal growth factor receptor (EGFR), and to explore the underlying signal transduction pathways. To this end, we constructed the LIRI cell model and mouse models, separately. According to RT-qPCR and Western blotting (WB) analysis results, miR-141 up-regulation together with ß-catenin and EGFR down-regulation within mouse pulmonary microvascular endothelial cells (PMVECs) or lung tissues was related to lung IRI. Besides, we conducted dual-luciferase reporter assay, which suggested the binding of EGFR to miR-141. In addition, we carried out TUNEL staining, HE staining, and flow cytometric analysis to assess the apoptosis of PMVECs and the injury to mouse lung tissues. Furthermore, we performed light-chain immunofluorescence assay to examine autophagosomes within PMVECs. According to our results, miR-141 suppressed ß-catenin level through reducing EGFR level. Besides, the miR-141/EGFR/ß-catenin axis enhanced autophagy to aggravate LIRI. To sum up, miR-141 suppresses EGFR expression to inhibit ß-catenin level, which subsequently aggravates autophagy and complicates LIRI. The above results offer the candidate therapeutic target for the treatment of lung IRI.


Subject(s)
MicroRNAs , Reperfusion Injury , Animals , Apoptosis/genetics , Autophagy/genetics , Endothelial Cells/metabolism , ErbB Receptors/genetics , Lung/metabolism , Mice , MicroRNAs/metabolism , Reperfusion Injury/genetics , Reperfusion Injury/metabolism , beta Catenin/genetics , beta Catenin/metabolism
8.
Int J Nanomedicine ; 17: 1483-1494, 2022.
Article in English | MEDLINE | ID: mdl-35378882

ABSTRACT

Ventilator-associated pneumonia (VAP) is a common and serious nosocomial infection in mechanically ventilated patients, increasing mortality, prolonging the patient length of stay, and increasing costs. In recent years, extensive studies on ventilator-associated pneumonia have shown that tracheal intubation plays an essential role in the pathogenesis of VAP, with the primary mechanism being the rapid colonization of the tracheal intubation surface by microbiota. Antibiotics do not combat microbial airway colonization, and antimicrobial coating materials offer new ideas to solve this problem. This paper reviews the current research progress on the role of endotracheal tube (ET) biofilms in the pathogenesis of VAP and antimicrobial coating materials.


Subject(s)
Microbiota , Pneumonia, Ventilator-Associated , Anti-Bacterial Agents/pharmacology , Humans , Intubation, Intratracheal/adverse effects , Pneumonia, Ventilator-Associated/etiology , Pneumonia, Ventilator-Associated/prevention & control , Respiration, Artificial/adverse effects
9.
J Cardiovasc Transl Res ; 15(3): 560-570, 2022 06.
Article in English | MEDLINE | ID: mdl-34599486

ABSTRACT

Cardiac fibrosis is a difficult clinical puzzle without effective therapy. Exosomes play an important role in alleviating cardiac fibrosis via angiogenesis. This research aimed to assess the effect of bovine milk on cardiac fibrosis. The proangiogenic effect of bovine milk exosomes was analyzed both in isoproterenol (ISO)-induced cardiac fibrosis rats in vivo and in human umbilical vein endothelial cells (HUVECs) after oxygen and glucose deprivation (OGD) in vitro. Results indicated that bovine milk exosomes alleviated the extracellular matrix (ECM) deposition and enhanced the cardiac function in cardiac fibrosis rat. The proangiogenic growth factors were significantly enhanced in rats accepted bovine milk exosomes. Meanwhile, bovine milk exosomes ameliorated the motility, migration, and tube-forming ability of HUVECs after OGD in vitro. Bovine milk exosomes alleviate cardiac fibrosis and enhance cardiac function in cardiac fibrosis rats via enhancing angiogenesis. Bovine milk exosomes may represent a potential strategy for the treatment of cardiac fibrosis.


Subject(s)
Exosomes , Myocardium , Neovascularization, Physiologic , Animals , Cattle , Exosomes/metabolism , Fibrosis , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Milk/metabolism , Myocardium/pathology , Rats
10.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-955475

ABSTRACT

Alzheimer's disease(AD)represents the main form of dementia;however,valid diagnosis and treatment measures are lacking.The discovery of valuable biomarkers through omics technologies can help solve this problem.For this reason,metabolomic analysis using ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS)was carried out on plasma,hippocampus,and cortex samples of an AD rat model.Based on the metabolomic data,we report a multi-factor combined biomarker screening strategy to rapidly and accurately identify potential bio-markers.Compared with the usual procedure,our strategy can identify fewer biomarkers with higher diagnostic specificity and sensitivity.In addition to diagnosis,the potential biomarkers identified using our strategy were also beneficial for drug evaluation.Multi-factor combined biomarker screening strategy was used to identify differential metabolites from a rat model of amyloid beta peptide 1-40(Aβ1-40)plus ibotenic acid-induced AD(compared with the controls)for the first time;lysophosphati-dylcholine(LysoPC)and intermediates of sphingolipid metabolism were screened as potential bio-markers.Subsequently,the effects of donepezil and pine nut were successfully reflected by regulating the levels of the abovementioned biomarkers and metabolic profile distribution in partial least squares-discriminant analysis(PLS-DA).This novel biomarker screening strategy can be used to analyze other metabolomic data to simultaneously enable disease diagnosis and drug evaluation.

11.
J Pharm Anal ; 11(5): 588-595, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34765271

ABSTRACT

From a regulatory perspective, drug quality consistency evaluation must concern different processes used for the same drug. In this study, an assessment strategy based on quality by design (QbD) was developed for population pharmaceutical quality evaluation. A descriptive analysis method based on QbD concept was first established to characterize the process by critical evaluation attributes (CEAs). Then quantitative analysis method based on an improved statistical process control (SPC) method was established to investigate the process indicators (PIs) in the process population, such as mean distribution, batch-to-batch difference and abnormal quality probability. After that rules for risk assessment were established based on the SPC limitations and parameters. Both the SPC parameters of the CEAs and the risk of PIs were visualized according to the interaction test results to obtain a better understanding of the population pharmaceutical quality. Finally, an assessment strategy was built and applied to generic drug consistency assessment, process risk assessment and quality trend tracking. The strategy demonstrated in this study could help reveal quality consistency from the perspective of process control and process risk, and further show the recent development status of domestic pharmaceutical production processes. In addition, a process risk assessment and population quality trend tracking provide data-based information for approval. Not only can this information serve as a further basis for decision-making by the regulatory authority regarding early warnings, but it can also reduce some avoidable adverse reactions. With continuous addition of data, dynamic population pharmaceutical quality is meaningful for emergencies and decision-making regarding drug regulation.

12.
J Int Med Res ; 49(3): 300060521999768, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33752447

ABSTRACT

OBJECTIVE: To assess safety and efficacy of a novel intubation laryngeal mask airway (ILMA) during the recovery period following supratentorial tumour surgery. METHODS: Patients who underwent supratentorial tumour surgery at our centre from January 2012 to December 2016 were eligible for this prospective randomised, parallel group study. We developed a novel ILMA using closely fitting laryngeal masks (No. 4/5) with 7.0/7.5 mm endotracheal tubes (ETT) plus screw fixators and anti-pollution sleeves. RESULTS: In total, 100 patients were intubated with the novel ILMA and 100 the ETT. There were no differences between groups in haemodynamic variables, oxygen saturation, exhaled CO2, or bispectral index all recorded during the 72-hour recovery period. However, there were significantly fewer incidences of coughing, less fluid drainage and lower haemoglobin levels in surgical fluid in the ILMA group compared with the ETT group. CONCLUSION: Our novel ILMA device was associated with reduced coughing, fluid drainage and blood in surgical drain during the recovery period following supratentorial tumour surgery.


Subject(s)
Laryngeal Masks , Supratentorial Neoplasms , Cough , Humans , Intubation, Intratracheal , Prospective Studies , Supratentorial Neoplasms/surgery
13.
Anal Chem ; 93(3): 1480-1488, 2021 01 26.
Article in English | MEDLINE | ID: mdl-33356172

ABSTRACT

A novel, effective, and label-free electrochemical sensor was constructed for investigating the interactions between cancer cells and molecules, based on targeted cancer cells immobilized on a bilayer architecture of N-doped graphene-Pt nanoparticles-chitosan (NGR-Pt-CS) and polyaniline (PANI). The interactions between folic acid (FA, positive control) and dimethyl sulfoxide (DMSO, negative control) and the choice of targeted cells, HepG2 and A549 cells, were investigated by measuring the current change of the sensor to [Fe(CN)6]3-/4- before and after interactions, and the binding constants were calculated to be 1.37 × 105 and 1.92 × 105 M-1 by sensing kinetics. Furthermore, 18 main components from Aidi injection (ADI) were studied to screen compounds that have interactions with different targeted cancer cells including HepG2 and A549 cells. The potential target groups of the interactions between screened active compounds and targeted cancer cells were analyzed through computer-aided molecular docking. In this sensing system, molecules did not require electrochemical activity, and different targeted cancer cells could be immobilized on the modified electrode surface, truly reflecting the categories and numbers of targets. Additionally, the proposed sensor specifically circumvented the current paradigm in most cells-based electrochemical sensors for screening drugs, in which the changes in cell behavior induced by drugs are monitored. This study provided a novel, simple, and generally applicable method for exploring the interaction of molecules with cancer cells and screening multitarget drugs.


Subject(s)
Antineoplastic Agents/chemistry , Biosensing Techniques , Dimethyl Sulfoxide/chemistry , Electrochemical Techniques , Folic Acid/chemistry , Aniline Compounds/chemistry , Chitosan/chemistry , Drug Evaluation, Preclinical , Graphite/chemistry , Humans , Metal Nanoparticles/chemistry , Molecular Docking Simulation , Particle Size , Platinum/chemistry , Surface Properties , Tumor Cells, Cultured
14.
Talanta ; 222: 121425, 2021 Jan 15.
Article in English | MEDLINE | ID: mdl-33167195

ABSTRACT

Folate receptors (FRs) are a class of valuable therapeutic target which is highly expressed on a variety of cancers. The accurate detection of the expression of FRs in different cells is conducive to improve the accuracy of FR targeted tumor therapy. Herein, a method based on nonimmobilized cell capillary electrophoresis (NICCE) combined with a mathematic model to quantify FRs on each single tumor cell was developed. At first, we studied the interactions between FA and A549, HT-29, HepG2, and U87MG cells by NICCE respectively, and calculated the kinetic parameters (Ka, k', ka, and kd). Next, we established a mathematic model to accurately determine the number of moles of FRs on per A549, HT-29, HepG2, and U87MG cell for the first time, that were (10.44 ± 0.53) × 10-19 mol, (34.32 ± 1.33) × 10-19 mol, (337.14 ± 10.11) × 10-19 mol, and (37.31 ± 2.13) × 10-19 mol. Then, these re-sults were proved to be consistent with the results of enzyme-linked immunosorbent assay (ELISA). Therefore, this method is simple, rapid, sensitive, and without protein separation or purification, which is expected to achieve clinical detection of cell membrane receptor expression level of cell membrane receptors on a single cell, which may be greatly beneficial to further clinical diagnosis and therapy.


Subject(s)
Neoplasms , Receptors, Cell Surface , Electrophoresis, Capillary , Folate Receptors, GPI-Anchored , Folic Acid , Humans , Models, Theoretical
15.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-908778

ABSTRACT

From a regulatory perspective,drug quality consistency evaluation must concern different processes used for the same drug.In this study,an assessment strategy based on quality by design(QbD)was developed for population pharmaceutical quality evaluation.A descriptive analysis method based on QbD concept was first established to characterize the process by critical evaluation attributes(CEAs).Then quantitative analysis method based on an improved statistical process control(SPC)method was established to investigate the process indicators(PIs)in the process population,such as mean distri-bution,batch-to-batch difference and abnormal quality probability.After that rules for risk assessment were established based on the SPC limitations and parameters.Both the SPC parameters of the CEAs and the risk of PIs were visualized according to the interaction test results to obtain a better understanding of the population pharmaceutical quality.Finally,an assessment strategy was built and applied to generic drug consistency assessment,process risk assessment and quality trend tracking.The strategy demon-strated in this study could help reveal quality consistency from the perspective of process control and process risk,and further show the recent development status of domestic pharmaceutical production processes.In addition,a process risk assessment and population quality trend tracking provide data-based information for approval.Not only can this information serve as a further basis for decision-making by the regulatory authority regarding early warnings,but it can also reduce some avoidable adverse reactions.With continuous addition of data,dynamic population pharmaceutical quality is meaningful for emergencies and decision-making regarding drug regulation.

16.
Talanta ; 207: 120259, 2020 Jan 15.
Article in English | MEDLINE | ID: mdl-31594574

ABSTRACT

We report a new method: biomimetic cell-cell adhesion capillary electrophoresis (BCCACE) to screen drugs targeting interactions between cell membrane receptors and ligands under an environment close to physiological conditions, in which the cell membrane receptors/ligands can maintain their natural conformations and bioactivity without being isolated and purified. Firstly, we screened twenty-one lactose derivatives by cell-immobilized capillary electrophoresis and obtained Gu-4 with the best activity (K = 3.58 ±â€¯0.22 × 104) targeting macrophage antigen-1 (Mac-1). Then, BCCACE was performed as follows: HEK 293 cells overexpressed with receptor (intercellular adhesion molecules-1, ICAM-1) were cultured and immobilized on the inner wall of capillaries as stationary phase, which simulated the endothelial cells lining on the inner surface of blood vessels. HEK 293 cells overexpressed with ligand Mac-1 as samples were used to simulate the neutrophils cells in blood vessels. And Gu-4 added into the running buffer solution as the antagonist was used to simulate the drug in blood. The results showed that Gu-4 (40 µM) could selectively inhibit cell-cell adhesion by targeting the interaction between Mac-1 and ICAM-1. Finally, the pharmaceutical efficacy assays of Gu-4 at cellular and animal levels were carried out using the concentration of 40 µM and the dose of 20 mg kg-1 respectively, which showed the anti-cancer metastasis activity of Gu-4 and the validity of the method. This method simulated a complete three-dimensional vascular model, which can easily obtain the suitable blood concentration of drugs. This system simulated the interaction between leukocytes and vascular endothelial cells in the bloodstream antagonized by drugs, and obtained the effective concentration of the antagonist. It can be used as an accuracy and efficient drug screening method and will be expected to become a new method to screen drugs targeting cell-cell adhesion.


Subject(s)
Biomimetics/methods , Cell Adhesion/drug effects , Drug Evaluation, Preclinical/methods , Electrophoresis, Capillary/methods , Glutamine/analogs & derivatives , Lactose/analogs & derivatives , Membrane Proteins/metabolism , Dose-Response Relationship, Drug , Glutamine/pharmacology , HEK293 Cells , Humans , Lactose/pharmacology , Ligands , Protein Binding/drug effects , Wound Healing/drug effects
17.
Anal Chim Acta ; 1063: 47-56, 2019 Jul 31.
Article in English | MEDLINE | ID: mdl-30967185

ABSTRACT

We reported a novel strategy for rapidly and accurately screening biomarkers based on ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) metabolomics data. First, the preliminary variables were obtained by screening the original variables using method validation. Second, the variables were selected from the preliminary variables and formed the variable sets by testing different thresholds of single factor (variable importance in projection (VIP), fold change (FC), the area under the receiver operating characteristic curve (AUROC), and -ln(p-value)). Then the partial least squares-discriminant analysis (PLS-DA) models were performed. The best threshold of each factor, and the corresponding variable set were found by comparing the models' R2X, R2Y, and Q2. Third, the second-step-obtained variable sets were further screened by multi-factors. The best combination of the multi-factors, and the corresponding variable set were found by comparing R2X, R2Y, and Q2. The expected biomarkers were thus obtained. The proposed strategy was successfully applied to screen biomarkers in urine, plasma, hippocampus, and cortex samples of Alzheimer's disease (AD) model, and significantly decreased the time of screening and identifying biomarkers, improved the R2X, R2Y, and Q2, therefore enhanced the interpreting, grouping, and predicting abilities of the PLS-DA model compared with generally-applied procedure. This work can provide a valuable clue to scientists who search for potential biomarkers. It is expected that the developed strategy can be written as a program and applied to screen biomarkers rapidly, efficiently and accurately.


Subject(s)
Alzheimer Disease/metabolism , Cerebral Cortex/metabolism , Chromatography, High Pressure Liquid , Hippocampus/metabolism , Mass Spectrometry , Metabolomics , Alzheimer Disease/diagnosis , Animals , Biomarkers/analysis , Biomarkers/metabolism , Cerebral Cortex/chemistry , Hippocampus/chemistry , Least-Squares Analysis , Male , Rats , Rats, Sprague-Dawley
18.
Anal Chim Acta ; 1045: 152-161, 2019 Jan 03.
Article in English | MEDLINE | ID: mdl-30454570

ABSTRACT

As there are more target categories on tumor cells/tissues than on receptor-overexpressing cells, and tumor tissues can better simulate TME, we established a new method of screening multi-target antitumor drugs by nonimmobilized tumor cells/tissues capillary electrophoresis under approximately tumor physiological environment. In this method, the natural structure and active conformation of the target proteins on tumor cells/tissues can be well maintained without separation and purification. Therefore, we successfully used this method to study the interactions between the Aidi injection (ADI)/its main components and tumor cells/tissues by optimizing a series of experimental conditions, discovered seven components with binding activity to A549 cells, five of them with specific interaction to tumor tissues, and calculated the binding kinetic parameters (K, ka, kd, and k'). Then, antitumor activity assays in vitro and in vivo were carried out to discover a new drug combination with higher targeting, better pharmaceutical efficacy, and lower toxic side effects. Finally, molecular docking studies were performed to investigate the potential target groups of the interactions between the effective drug combination and A549 cells/tissues. In summary, the method was verified to be valid and feasible, and can be easily transferred to a capillary array electrophoresis for high-throughput drug screening.


Subject(s)
Adenocarcinoma of Lung/pathology , Antineoplastic Agents/analysis , Electrophoresis, Capillary , Lung Neoplasms/pathology , Adenocarcinoma of Lung/drug therapy , Antineoplastic Agents/therapeutic use , Cell Line, Tumor , Drug Evaluation, Preclinical , Humans , Lung Neoplasms/drug therapy
19.
Analyst ; 143(20): 4981-4989, 2018 Oct 08.
Article in English | MEDLINE | ID: mdl-30225497

ABSTRACT

In this research a method called immobilized cell capillary electrophoresis (ICCE) was established under approximately physiological conditions for rapid screening of anti-tumor metastasis drugs targeting integrin macrophage antigen-1 (MAC-1). In this method, separation and purification of the target receptors on cell membranes was unnecessary, thus, maintaining their natural conformation and bioactivity. MAC-1-, CD11b-, or CD18-overexpressing HEK293 cells (human embryonic kidney) were cultured and immobilized on the inner wall of capillaries as stationary phase, and their interactions with lactosyl derivative Gu-4 (positive control)/dimethylsulfoxide (DMSO; negative control) were studied using ICCE. Using this method, 29 phenylethanoid glycosides from Cistanches Herba were screened, and the binding kinetic parameters (K, ka, kd, and k') of active compounds were calculated, and the specific subunits of MAC-1 were determined. Then, molecular docking studies were performed to discover the direct interaction sites between active compounds and MAC-1, and the order of Glide-calculated Emodel value obtained from the molecular docking study is consistent with that of the binding constants obtained using ICCE. Finally, pharmaceutical efficacy assays in vitro and in vivo were carried out to show that the anti-tumor metastasis activity of the active compound had better pharmaceutical efficacy and lower toxic side effects. The method was verified to be valid and practical for further use, and it is expected that it will be transferred to capillary array electrophoresis for use in high-throughput drug screening.


Subject(s)
Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Electrophoresis, Capillary/methods , Glycosides/pharmacology , Macrophage-1 Antigen/metabolism , Neoplasm Metastasis/prevention & control , A549 Cells , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/metabolism , Cells, Immobilized/metabolism , Cistanche/chemistry , Drug Screening Assays, Antitumor/methods , Glycosides/chemistry , Glycosides/metabolism , HEK293 Cells , Humans , Kinetics , Macrophage-1 Antigen/chemistry , Mice, Inbred C57BL , Molecular Docking Simulation , Protein Binding
20.
Biosens Bioelectron ; 116: 81-88, 2018 Sep 30.
Article in English | MEDLINE | ID: mdl-29860090

ABSTRACT

Herein, the ternary composites, ultrasmall metal nanoparticles encapsulated in the anionic metal-organic frameworks/electrochemically reduced graphene oxide (MNPs@Y-1, 4-NDC-MOF/ERGO, M = Ag, Cu) are constructed by a cationic exchange strategy and an electrochemical reduction process for the electrochemical determination of H2O2. Both AgNPs@Y-1, 4-NDC-MOF/ERGO and CuNPs@Y-1, 4-NDC-MOF/ERGO display excellent electrocatalytic activity toward H2O2, but the former is superior to the latter. Such a difference in electrocatalytic activity can be explained by the characterization measurements, and the results manifest MNPs@Y-1, 4-NDC-MOF/ERGO (M = Ag, Cu) electrocatalysts have subequal MNPs sizes and electrochemical surface areas, but different electron transfer rate constants. The AgNPs@Y-1, 4-NDC-MOF/ERGO sensor shows a linear detection range from 4 to 11,000 µM with the detection limit of 0.18 µM. Moreover, MNPs@Y-1, 4-NDC-MOF/ERGO (M = Ag, Cu) exhibit excellent anti-interference performance and can be used for the detection of H2O2 released from living cells. The proposed sensor takes full advantage of the catalytic property of MNPs, the size selectivity of Y-1, 4-NDC-MOF, and the fast electron transport effect of ERGO. Thus, the MNPs@Y-1, 4-NDC-MOF/ERGO/GCE (M = Ag, Cu) can be utilized to detect oxidase activities by monitoring H2O2 produced in the presence of substrate and oxidase, and it is expected that composites with the molecular sieving effect and catalytic activity can be widely applied for catalysis, biomedicine, and biosensing fields.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Hydrogen Peroxide/analysis , Metal-Organic Frameworks/chemistry , A549 Cells , Anions/chemistry , Catalysis , Copper/chemistry , Electrodes , Graphite/chemistry , Humans , Hydrogen Peroxide/chemistry , Limit of Detection , Metal Nanoparticles/chemistry , Metal-Organic Frameworks/chemical synthesis , Oxides/chemistry , Particle Size , Silver/chemistry , Yttrium/chemistry
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