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Molecular imprinting-based surface-enhanced Raman scattering (MI-SERS) sensors have shown remarkable potential from an academic standpoint. However, their practical applications, especially in the detection of large-size protein (≥10 nm), face challenges due to the lack of versatile sensing strategies and nonspecific fouling of matrix species. Herein, we propose a Raman reporter inspector mechanism (RRIM) implemented on a protein-imprinted polydopamine (PDA) layer coated on the SERS active substrate. In the RRIM, after large-size protein recognition, the permeability of the PDA imprinted cavities undergoes changes that are scrutinized by Raman reporter molecules. Target proteins can specifically bind and fully occupy the imprinted cavities, whereas matrix species cannot. Then, Raman reporter molecules with suitable size are introduced to serve as both inspectors of the recognition status and inducers of the SERS signal, which can only penetrate through the vacant and nonspecifically filled cavities. Consequently, changes in the SERS signal exclusively originate from the specific binding of target proteins, while the nonspecific recognition of matrix species is curbed. The RRIM enables reproducible quantitation of the large-size cyanobacteria-specific protein model (≥10 nm), phycocyanin, at the level down to 2.6 × 10-3 µg L-1. Finally, the practical applicability of the RRIM is confirmed by accurately analyzing crude urban waterway samples over 21 min without any pretreatment.
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Milk is a naturally complex medium that is suitable for the growth of most lactic acid bacteria. Unfortunately, Lactiplantibacillus plantarum ST-III grow poorly in milk without supplementation. To solve this problem, we use fresh pineapple and mung beans juice to develop an edible proliferator for L. plantarum ST-III. Our comparative analysis of metabolomics changes before and after fermentation reveals that amino acids and dipeptides are the most consumed compounds, with other substances including nucleotides and vitamins, implying the mechanism of proliferation. Combining the KEGG metabolic pathway analysis, substances that may promote the growth of L. plantarum ST-III in milk were screened. To explore which component of the proliferator is required for L. plantarum ST-III cultivate, we supplemented with several combinations of molecules aforementioned in milk. The simulation addition experiment results of L. plantarum ST-III in milk show that if any additions are missing, the concentration of viable bacteria is lower. Only when it contains all additives can the highest concentration of viable bacteria be obtained. Compared with the control, the fold change of the viable bacteria is about 32. Thus, it proves that milk primarily lacked available amino acids, dipeptides, uracil, xanthine, nicotinamide, and manganese for the growth of L. plantarum ST-III.
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OBJECTIVE: The purpose of this study was to compare the biomechanical and clinical results of two surgical methods for the treatment of vertical femoral neck fractures: Femoral neck system (FNS) and traditional three cannulated cancellous screws (CCS). METHODS: First, we developed three different vertical femoral neck fracture models for the finite element analysis, with angles of 55°, 65°, and 75°, respectively. Two experimental groups were set up: the FNS group and the CCS group. Each fracture group was tested under axial loads of 2100 N to measure the femur's displacement, Von Mises stress (VMS), and its internal fixation components. Secondly, we retrospectively included the cases of vertical femoral neck fractures with FNS and CCS in our hospital from May 2019 to May 2021. In this study, we compared the duration of intraoperative fluoroscopy, operative time, hospital stay, fracture healing time, Hemoglobin loss, Harris score of hip joint function, and postoperative complications among patients undergoing hip joint replacement. RESULTS: In terms of finite element analysis, FNS has better anti-displacement stability than CCS at 55°and 65°, while FNS is greater than CCS in Von Mises stress. Clinically, we followed up on 87 patients for an average of 12 months. FNS was superior to traditional CCS in fracture healing time, operation time, fluoroscopy duration, fracture healing time, and Harris hip function score. CONCLUSION: FNS is superior to traditional CCS in biomechanical and clinical aspects of treating vertical femoral neck fractures. There is potential for FNS to become a new treatment option for vertical femoral neck fractures.
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Fracturas del Cuello Femoral , Cuello Femoral , Humanos , Análisis de Elementos Finitos , Estudios Retrospectivos , FluoroscopíaRESUMEN
OBJECTIVE: Analysis of the risk factors affecting hip function and complications after femoral neck system (FNS) surgery for femoral neck fractures is of great significance for improving the procedure's efficacy. METHODS: The data of patients with femoral neck fractures who underwent FNS surgery in our hospital between October 2019 and October 2020 were retrospectively analyzed. Age, gender, time from injury to operation, fracture classification, operation time, fracture reduction, and postoperative weight-bearing time information were set as potential factors that may affect the results. Hip Harris scores were performed at 12 months postoperatively, and postoperative complication data (e.g., femoral head necrosis, nonunion, and femoral neck shortness) were collected. The risk factors affecting hip function and complications after FNS surgery were predicted using linear and logistic regression analyses. RESULTS: A total of 69 cases of femoral neck fracture were included, with an average age of 56.09 ± 11.50 years. The linear analysis demonstrated that the age and fracture type of the patients were the risk factors affecting the Harris score of the hip joint after FNS surgery. Older patients with displaced femoral neck fractures had an inferior postoperative hip function. In addition, fracture type, reduction of the femoral neck, and postoperative weight-bearing significantly impacted postoperative complications. Displaced fractures, negative fixation, and premature weight-bearing (< 6 weeks) were risk factors for postoperative complications. The Harris score of patients with a shortened femoral neck in the included cases was not significantly different from that of patients without shortening (P = 0.25). CONCLUSIONS: Advanced age and fracture type are important evaluation indicators of the Harris score after FNS internal fixation of femoral neck fractures in young patients. Fracture type, fracture reduction, and postoperative weight-bearing time are risk factors for complications after FNS.
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Fracturas del Cuello Femoral , Cuello Femoral , Humanos , Adulto , Persona de Mediana Edad , Anciano , Estudios Retrospectivos , Fracturas del Cuello Femoral/cirugía , Factores de Riesgo , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/etiología , Articulación de la Cadera/diagnóstico por imagen , Articulación de la Cadera/cirugíaRESUMEN
Objective: This study aimed to evaluate the clinical efficacy of an enhanced minimally invasive NICE joint technique combined with dual adjustable loop steel plate internal fixation for treating acute acromioclavicular joint dislocation. Methods: A retrospective analysis was conducted on 63 surgical patients treated with acute acromioclavicular joint dislocation from May 2017 to March 2022. Among them, 33 cases were treated with the clavicle hook plate, and 30 cases were treated with the minimally invasive loop plate. We compared hospitalization duration, incision length, surgical duration, intraoperative bleeding, visual analogue pain scale scores, shoulder joint Constant scores at 6 months before and after surgery, and the incidence of complications between the two groups. Results: The comparison between the two groups, including hospitalization duration, incision length, surgical duration, intraoperative bleeding volume, and shoulder joint Constant score at 6 months post-surgery, revealed statistically significant differences where the loop plate group had better results. One case (1/33) experienced postoperative complications in the hook plate group, including screw loosening and plate failure. Additionally, there were 8 cases (8/33) of subacromial osteolysis, 10 cases (10/33) of acromial impact, and 5 cases (5/33) of residual shoulder pain. Conversely, only 1 case (1/30) in the loop plate group had residual shoulder pain. Conclusions: The surgical technique involving the reconstruction of the coracoclavicular ligament using an enhanced minimally invasive NICE junction combined with double adjustable loop steel plate placement in the clavicular small bone canal is characterized by simplicity, safety, minimal invasiveness, excellent functional recovery, fewer complications, and superior clinical efficacy compared to clavicular hook steel plates.
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Articulación Acromioclavicular , Luxaciones Articulares , Luxación del Hombro , Humanos , Luxaciones Articulares/cirugía , Estudios Retrospectivos , Articulación Acromioclavicular/cirugía , Dolor de Hombro , Luxación del Hombro/cirugía , Resultado del Tratamiento , AceroRESUMEN
Microneedles (MNs) are currently one of the most promising tools for skin interstitial fluid (ISF)-based biosensing, while it is still a challenge to expand the detectable biomarkers in ISF due to limited MNs types and detection techniques. Herein, highly sensitive internal-standard surface-enhanced Raman scattering microneedles (IS-SERS-MNs) were developed, which enabled the reliable detection of bacterial metabolites in ISF as new detectable biomarkers for infection diagnosis. The developed IS-SERS-MNs can not only directly detect pyocyanin (a representative bacterial metabolite) present in mouse dermal ISF but also indirectly detect pyocyanin in the hypodermis via its diffusion into the dermis, revealing a new possible pathway for the source of biomarkers in dermal ISF. Moreover, the SERS signal of pyocyanin was also clearly detected at real mouse wounds, indicating that the developed IS-SERS-MNs have great potential in minimally invasive and painless diagnosis of bacterial infection via a new ISF route. This work not only develops IS-SERS-MNs as a powerful tool for expanding the application of SERS-based MNs but also provides a new chance for ISF-related infection diagnosis.
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Líquido Extracelular , Espectrometría Raman , Ratones , Animales , Líquido Extracelular/metabolismo , Agujas , Piocianina , Piel/metabolismo , Biomarcadores/metabolismoRESUMEN
BACKGROUND: Clinical angiography and vascular microperfusion confirmed that the femoral head retains blood supply after a collum femur fracture. However, no animal model accurately mimics this clinical situation. This study was performed to establish a rat model with retained viability of the femoral head and partial vasculature deprivation-induced traumatic caput femoris necrosis by surgery. METHODS: Thirty rats were randomly divided into three groups (n = 10 per group): normal group, sham-operated group (Control), and ischemic osteonecrosis group. The femoral head of the normal group of rats underwent a gross anatomy study and microangiography to identify femoral head blood supply. Microsurgical techniques were used to cauterize the anterior-superior retinacular vessels to induce osteonecrosis. Hematoxylin and Eosin (H&E) staining were used for femoral head histologic assessment. Morphologic assessments of the deformity in and trabecular bone parameters of the femoral head epiphysis were performed using micro-CT. RESULTS: The blood supply of the femoral head in rats primarily came from the anterior-superior, inferior, and posterior retinacular arteries. However, anterior-superior retinacular vasculature deprivation alone was sufficient in inducing femoral head osteonecrosis. H&E showed bone cell loss in nuclear staining, disorganized marrow, and trabecular structure. The bone volume (BV) decreased by 13% and 22% in the ischemic group after 5 and 10 weeks, respectively. The mean trabecular thickness (Tb.Th) decreased from 0.09 to 0.06 mm after 10 weeks. The trabecular spacing (Tb.Sp) increased from 0.03 to 0.05 mm after 5 weeks, and the epiphyseal height-to-diameter (H/D) ratio decreased. CONCLUSIONS: We developed an original and highly selective rat model that embodied femoral head traumatic osteonecrosis induced by surgical anterior-superior retinacular vasculature deprivation.
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Necrosis de la Cabeza Femoral , Traumatismos de los Tejidos Blandos , Animales , Epífisis/cirugía , Fémur/patología , Cabeza Femoral/irrigación sanguínea , Cabeza Femoral/diagnóstico por imagen , Cabeza Femoral/cirugía , Necrosis de la Cabeza Femoral/diagnóstico por imagen , Necrosis de la Cabeza Femoral/etiología , Necrosis de la Cabeza Femoral/cirugía , Humanos , Ratas , Traumatismos de los Tejidos Blandos/patología , Microtomografía por Rayos XRESUMEN
Understanding the in vivo behavior of photothermal nanomedicines (PTNMs) is important for drug development and evaluation. However, it is still very challenging. Herein, two key parameters, i.e., the depth of PTNMs under biological tissue and the drug release ratio of PTNMs in vivo, can be revealed by a near-infrared (NIR) light-responsive surface-enhanced Raman scattering (SERS) strategy. The fabricated PTNMs were composed of waxberry-like gold nanoparticles, model drug curcumin, and an elaborately selected NIR light-responsive Raman reporter (3,3'-diethylthiatricarbocyanine iodide, DTTC). The response mechanism of DTTC to NIR light was investigated as photodegradation. NIR light irradiation heated the gold nanoparticles, triggered the release of a model drug, and simultaneously decreased the SERS intensity of the PTNMs. In vitro experiment results revealed that the SERS intensity decrease could well reflect the depth of PTNMs with a correlation coefficient of more than 0.99. On this basis, after in situ SERS detection, the depth of PTNMs in a tumor could be revealed with satisfactory accuracy. Moreover, the decrease in the SERS intensity of PTNMs showed a highly similar trend to the increase in the drug release, suggesting that it could be used for real-time monitoring of drug release of PTNMs. This study not only opens a new avenue for the release study of many inactive fluorescent and Raman drugs of PTNMs but also provides an effective way for reporting the depth, which greatly promotes the application of PTNMs in vivo.
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Oro , Nanopartículas del Metal , Liberación de Fármacos , Nanomedicina , Espectrometría RamanRESUMEN
For periprosthetic joint infection (PJI) patients, an early and rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) in joint synovial fluid is of great significance for receiving timely treatment and avoiding side effects. In clinical practice, the methods for detecting MRSA include the culture-based method and the PCR-based mecA gene detection method with fluorescent readout. However, the culture-based method requires up to 3-7 days for incubation and elaborative screening. The PCR-based molecular diagnosis, due to its high sensitivity, improves the detection time but sacrifices cost and gives false-positive results. Herein, a ligation chain reaction (LCR)-based electrochemical biosensor was developed to detect the mecA of MRSA with the advantages of rapidity, accuracy and low cost. In this system, an integrated dsDNA labeled with thiol and biotin at both terminals is generated only in the presence of the target DNA after LCR, followed by immobilization of the integrated dsDNAs on the bovine serum albumin (BSA)-coated gold electrode, and then the streptavidin horseradish peroxidase (SA-HRPs) is specifically bound to the biotin labels via biotin-streptavidin interaction, generating the catalytic amperometric readout. Impressively, the developed method achieved the detection of rare mecA in the joint synovial fluid of PJI patients (417-666 copies as quantified by qPCR). The proposed electrochemistry-based method is highly convenient for the point-of-care testing and was comparable with PCR in sensitivity, but superior in selectivity (single-base differentiation) and cost (nanomolar DNA probe consumption and simple device), demonstrating its huge potential in clinical applications for MRSA diagnosis.
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Técnicas Biosensibles , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Proteínas Bacterianas/genética , ADN , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Proteínas de Unión a las Penicilinas/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Infecciones Estafilocócicas/diagnóstico , Líquido SinovialRESUMEN
Osteosarcoma (OS), the most common malignant bone tumor with high metastatic potential, frequently affects children and adolescents. Epidermal growth factor receptor (EGFR)-targeted tyrosine kinase inhibitors exhibit encouraging anti-tumor activity for patients with solid tumors, whereas their effects on OS remain controversial. In the present study, we aimed to elucidate the anti-tumor activity of gefitinib for OS, as well as to explore the underlying mechanisms. Gefitinib inhibits cell viability, tumor growth, cell migration, and invasion and promotes cell apoptosis and G1 cycle arrest in OS at a relatively high concentration via suppressing the PI3K/Akt and ERK pathways. However, gefitinib treatment results in the feedback activation of signal transducer and activator of transcription 3 (STAT3) induced by interleukin 6 (IL-6) secretion. Combined treatment with gefitinib and stattic, an inhibitor for STAT3 phosphorylation, engenders more evident inhibitory effects on cell proliferation, migration, and invasion and promotive effects on cell apoptosis and G1 phase arrest in OS, compared with the single exposure to gefitinib or stattic. Western blot analysis demonstrates that stattic treatment in gefitinib-treated OS abrogates the IL-6-induced STAT3 activation and subsequently further restrains the activities of EGFR, Akt, and ERK pathways in tumor cells. This study confirms that the EGFR inhibitor of gefitinib has moderate anti-tumor effects on OS through IL-6 secretion-mediated STAT3 activation. Additional administration of stattic in EGFR-targeted therapies may contribute to improve the efficacy for OS.
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Antineoplásicos/farmacología , Óxidos S-Cíclicos/farmacología , Receptores ErbB/antagonistas & inhibidores , Interleucina-6/metabolismo , Osteosarcoma/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/farmacología , Factor de Transcripción STAT3/antagonistas & inhibidores , Animales , Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Óxidos S-Cíclicos/uso terapéutico , Femenino , Gefitinib/farmacología , Gefitinib/uso terapéutico , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Osteosarcoma/patología , Inhibidores de Proteínas Quinasas/uso terapéutico , Factor de Transcripción STAT3/metabolismo , Ensayo de Tumor de Célula Madre , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
This article introduces the safety risks of the novel light-based home-use hair removal device, and analyzes the differences in regulation among China, the United States and the European Union. In China, household intense pulsed light hair removal devices will also be supervised in accordance with medical device regulations. Therefore, the safety standards adopted in the absence of specific regulations are no longer applicable to the new regulatory requirements. It is imperative to adopt the new standards available to home photoepilators, so as to ensure the safety and effectiveness of the approved devices.
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Remoción del Cabello , China , Unión Europea , Estándares de Referencia , Estados UnidosRESUMEN
Surface-enhanced Raman resonant scattering (SERRS) tags encoded with near-infrared (NIR) Raman reporters showed great potential for in vivo detection owing to their ultrasensitivity. However, in vivo signal stability of such tags is a remaining problem due to the lack of suitable silica coating method because the weakly adsorbed NIR reporters tend to detach from traditional gold nanosubstrates in the ethanol-rich and high pH conditions, which are commonly used for silica coating. Herein, we propose a silica coating method for NIR SERRS tags by using waxberry-like gold nanoparticles (NPs) as substrates. The lipid bilayer of the NPs played a crucial role in the coating, which can encapsulate the NIR Raman reporter via hydrophobic interactions and prevent the interference from a harsh medium. Thus, the silica-coated tags well preserved ultrasensitivity of bare tags and simultaneously gained satisfactory signal stability in vivo. Moreover, the coating method is compatible for the encapsulation of a variety of thiol group-free NIR reporters (as exemplified by DTTC, Cy7, IR792, and DIR), relying on which a tag-pair with distinguishable peaks can be screened (labeling with DTTC and Cy7, respectively). In vivo duplexing detection revealed that the tag-pair-labeled liposome was cleared faster in the liver than polydopamine NPs within one mouse. The developed method paves an easy way for gaining high-quality SERRS tags and will promote their in vivo multiplex analysis and diagnostics applications.
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Materiales Biomiméticos/química , Colorantes/química , Rayos Infrarrojos , Magnoliopsida , Dióxido de Silicio/química , Espectrometría Raman/métodos , Animales , Oro/química , Concentración de Iones de Hidrógeno , Nanopartículas del Metal/química , Ratones , Compuestos de Sulfhidrilo/química , Propiedades de SuperficieRESUMEN
Proliferative vitreoretinopathy (PVR) is the most common cause of surgical failure in the rhegmatogenous retinal detachment (RD) treatment. Retinal pigment epithelial (RPE) cell proliferation, migration, and the synthesis of extracellular matrix (ECM) are intrinsic to the formation of a PVR membrane. High level of interleukin-6 (IL-6) has been found in the vitreous of PVR patients, while the role of IL-6 in RPE cells remaining further characterized. In the present study, we evaluated the potential regulatory effects of IL-6 on cell migration, ECM components, and transforming growth factor ß2 (TGF-ß2) expression in RPE cells. Furthermore, cell counting kit-8 (CCK8) assay was used to investigate cell proliferation activity. We found that IL-6 promoted fibronectin (Fn) and type I collagen (COL-1), TGF-ß2 expression in RPE cells, also stimulate RPE cell migration effectively. Moreover, the induction of IL-6 activated the Janus kinase/signal transducers and activators of transcription (JAK/STAT3) and the nuclear factor kappa-B (NF-κB) signaling pathways significantly. Simultaneously, both JAK/STAT3 and NF-κB pathways inhibitors, WP1066 and BAY11-7082, alleviated IL-6-induced biological effects, respectively. However, it was noted that IL-6 had little effect on α-smooth muscle actin (α-SMA) expression. Collectively, our results reveal that IL-6 promotes RPE cell migration and ECM synthesis via activating JAK/STAT3 and NF-κB signaling pathways, which may play a crucial role in PVR formation.
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Matriz Extracelular/metabolismo , Interleucina-6/metabolismo , Epitelio Pigmentado de la Retina/metabolismo , Movimiento Celular , Células Cultivadas , Humanos , Epitelio Pigmentado de la Retina/citologíaRESUMEN
Osteogenic differentiation (OD) of bone marrow mesenchymal stem cells (BMSCs) is critically important for mitigation of osteoporosis. Glucocorticoids (GCs) are extensively used for treating chronic inflammation, although long-term exposure to GCs is capable of triggering osteoporosis. microRNAs (miRNAs) have been reported to play a critical role in bone diseases. In the present study, we treated BMSCs with dexamethasone (DEX) during OD to stimulate GC-mediated osteoporosis. Microarray and quantitative polymerase chain reaction (Q-PCR) assays demonstrated that miR-199a was upregulated during OD of BMSCs, while DEX treatment caused a significant reduction in miR-199a. Alkaline phosphatase (ALP) activity, Alizarin red (AR) staining, and Q-PCR were applied to assess the role of miRNA-199a overexpression in DEX-triggered OD inhibition. miR-199a was able to rescue OD and ALP activity, which were inhibited by DEX. Additionally, we observed that ALP, BMP2, COL1A1, and Runx2 were increased after transfection of miRNA-199a mimics. Furthermore, we confirmed that miRNA-199a facilitates OD of BMSCs through direct inhibition of Klotho protein and messenger RNA expression affecting the downstream fibroblast growth factor receptor 1/extracellular-signal-regulated kinase and Janus kinase 1/signal transducer and activator of transcription 1 pathways. This study indicates that miR-199a plays a critical role in preventing GC-mediated osteoblast differentiation and may function as a promising miRNA biomarker for osteoporosis.
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Glucuronidasa/metabolismo , MicroARNs/genética , Osteogénesis/genética , Animales , Células de la Médula Ósea/metabolismo , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Dexametasona/efectos adversos , Dexametasona/farmacología , Femenino , Glucocorticoides/metabolismo , Glucocorticoides/farmacología , Proteínas Klotho , Masculino , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , MicroARNs/metabolismo , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Osteoporosis/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVES: Despite the important role of Galectin-9 (Gal-9) in inflammation and angiogenesis, only a few studies on Galectin-9 expression have been performed in rheumatoid arthritis (RA) patients. The present study aimed to identify the concentration of Galectin-9 in plasma, its expression in peripheral blood T cell subsets in RA patients, and the association between Galectin-9 and vascular endothelial growth factor (VEGF) in RA. METHODS: One hundred and five active RA patients and 52 age- and sex-matched healthy controls (HCs) were enrolled in the study. Gal-9, vascular endothelial growth factor (VEGF), and tumour necrosis factor (TNF)-α level in plasma were determined by ELISA. The intracellular positive expression rates and medium fluorescence intensity (MFI) of Gal-9 and VEGF in T cell subsets, were examined by flow cytometry. RESULTS: Plasma Gal-9, TNF-α, and VEGF levels were higher in the RA group than in the HCs group. The plasma Gal-9 level positively correlated with Gal-9 expression in the total lymphocyte and CD3+ T cell, CRP, SDAI, and CDAI of RA patients. The Gal-9 expressions in the cytoplasm of CD4+ T, CD8+ T, Treg, and DNT cells positively correlated with plasma TNF-α levels in RA patients. The Gal-9 expressions in CD4+ T and CD8+ T subsets also positively correlated with plasma VEGF levels. The plasma VEGF levels and VEGF expressions in CD4+ T, CD8+ T, and Treg subsets positively correlated with SDAI and CRP in RA patients. CONCLUSIONS: Gal-9 can be a potential biomarker for RA disease activity. Gal-9 is probably associated with angiogenesis processing in RA.
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Artritis Reumatoide , Factor A de Crecimiento Endotelial Vascular , Galectinas , Humanos , Subgrupos de Linfocitos T , Factor de Necrosis Tumoral alfaRESUMEN
Surface coating determined the sensitivity and stability of surface-enhanced Raman scattering (SERS) tags in bioanalysis. The reported various coatings suffered from the drawbacks of a lack of rigidity, stability, or synthesis versatility. Herein, we demonstrated robust polystyrene (PS) coated SERS tags that could be prepared by an easy and universal approach. Taking advantages of biocompatible, transparent, compact properties of PS shell, the coated tags showed satisfactory sensitivity, biocompatibility, and superior structural stability in cell and in vivo imaging applications. More importantly, the PS coating strategy allowed for the encapsulation of SERS tags encoded with not only thiolated but also nonthiolated Raman reporters without loss of sensitivity, as exemplified in the synthesis of 9 different resonant dye-encoded tags. Moreover, the coating of SERS tags with various kinds of substrates was achieved via the same standard protocol. Comparing with widespread silica coated tags, the PS coated ones were more stable in harsh conditions and had an easily expanded ultrasensitive (resonant) tags library with much lower cost (no need of expensive sulfhydryl/isothiocyano reporters with limited types), illustrating great promise as standard analytical tools of commercialized value for bioanalysis, medical diagnostics, and environmental science studies.
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Poliestirenos/química , Carbono/análisis , Tamaño de la Partícula , Péptidos/análisis , Proteínas/análisis , Dióxido de Silicio/análisis , Espectrometría Raman , Propiedades de Superficie , Titanio/análisisRESUMEN
BACKGROUND: Tripartite motif-containing protein 11 (TRIM11), a member of RING family of E3 ubiquitin ligases, is identified as an oncogene in certain human tumors. However, the detailed biological function of TRIM11 in chordoma is still unclear. The purpose of present research is to explore the role of TRIM11 in human chordoma cells. METHODS: TRIM11 was induced silencing and overexpression in human chordoma cells using RNA interference (RNAi) and lentiviral vector. qRT-PCR and western blot were used to determine gene expression in chordomas cells. Meanwhile, cell counting kit-8 (CCK-8) assay was used to examine the cell proliferation rate. Flow cytometry analysis was performed to quantify the cell apoptosis rate. RESULTS: We identified that TRIM11 was upregulated in chordomas tissues. Moreover, TRIM11 presented pro-proliferation and anti-apoptosis function in chordoma cells. Further, LY294002, a specific AKT inhibitor, was utilized to examine the connection between TRIM11 and AKT in human chordoma cells. Importantly, our findings elucidated that TRIM11 promoted the growth of chordoma cells and involved in AKT signaling. Much more importantly, knockdown of TRIM11 significantly upregulated the translation of PH domain leucine-rich repeats protein phosphatase 1 (PHLPP1), whereas did not affect its transcription. Results that obtained from co-immunoprecipitation (Co-IP) and ubiquitination assay demonstrated TRIM11 interacted with PHLPP1 and promoted its ubiquitination in chordoma cells. Moreover, overexpression of PHLPP1 inhibited the phosphorylation of AKT in human chordomas cells. These results suggested that TRIM11 mediated the post-translation modification of PHLPP1 and was a novel component in PHLPP1/AKT signaling pathway in human chordoma cells. CONCLUSIONS: Taken together, the present research not only enhanced the understanding of TRIM11 but also indicated its potential target and signaling pathway in human chordoma cells.Trial registration retrospectively registered.
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Hydrogen peroxide (H2O2), as a major component of reactive oxygen species (ROS), plays an important role in normal physiological processes. A H2O2 burst also occurs in the ischemia/reperfusion (I/R) process and causes a series of physiological and pathological injuries. Therefore, it is important to determine concentration fluctuations of H2O2. Here we develop a ratiometric fluorescent probe, Cy-ArB, which shows high selectivity and sensitivity toward H2O2. The fluorescence response of the probe is triggered by the reaction of borate esters with H2O2, and this process releases a near-infrared heptamethine cyanine fluorophore which has the ability of mitochondrial tracing. Hence, the probe can be used for real-time monitoring of H2O2 fluctuations in the mitochondrial respiration chain. Finally, we explore the fluctuations of H2O2 in cells and in vivo during the I/R process using the probe Cy-ArB. The results of our experiments prove that our probe is a potential candidate for clinical surgery pre-evaluation.
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Boratos/química , Colorantes Fluorescentes/química , Peróxido de Hidrógeno/análisis , Indoles/química , Daño por Reperfusión/fisiopatología , Animales , Apoptosis/fisiología , Boratos/síntesis química , Boratos/efectos de la radiación , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Células Hep G2 , Humanos , Peróxido de Hidrógeno/metabolismo , Indoles/síntesis química , Indoles/efectos de la radiación , Límite de Detección , Hígado/fisiopatología , Ratones Endogámicos BALB C , Microscopía Confocal/métodos , Microscopía Fluorescente/métodos , Mitocondrias/metabolismoRESUMEN
Maintaining the redox balance of biological systems is a key point to maintain a healthy physiological environment. Excessive iron ions (Fe3+) can cause apoptosis, tissue damage and death. Fortunately, ascorbic acid (AA) as a reducing agent has been evaluated for the reduction of Fe3+. Moreover, AA plays an important role in relieving hypoxia-induced oxidative stress. Therefore, the real-time imaging of the Fe3+ and AA fluctuations is important for understanding their biofunctions in cells and in vivo. In this work, we developed a fluorescent nanoprobe carbon dot-desferrioxamine B (CD-DB) by the conjugate connection of CDs and desferrioxamine B (a complexing agent for Fe3+) for the associated detection of Fe3+ and AA. CD-DB exhibited excellent sensitivity and selectivity for the detection of Fe3+ and AA. The nanoprobe CDs-DB@Fe obtained by the reaction of CD-DB and Fe3+ was suitable for tracing the dynamic changes of AA in cells and in vivo. Therefore, CDs-DB@Fe was used for monitoring the fluctuation of AA in hypoxic cell models, hypoxic zebrafish models and liver ischemia mice models. These results exhibited the decrease in AA under hypoxic conditions because AA was consumed to neutralize free radicals and relieve hypoxia-induced oxidative stress damage. The ideal biocompatibility and low toxicity make our nanoprobe a potential candidate for the research of the physiological effects of AA in vivo.
Asunto(s)
Ácido Ascórbico/análisis , Colorantes Fluorescentes/química , Hipoxia/metabolismo , Hierro/análisis , Puntos Cuánticos/química , Animales , Carbono/química , Hipoxia de la Célula , Deferoxamina/química , Colorantes Fluorescentes/síntesis química , Células Hep G2 , Humanos , Isquemia/metabolismo , Límite de Detección , Hígado/irrigación sanguínea , Hígado/metabolismo , Ratones Endogámicos BALB C , Microscopía Confocal/métodos , Microscopía Fluorescente/métodos , Estrés Oxidativo , Pez CebraRESUMEN
Cardiac troponin I (cTnI) was considered as the "gold standard" for acute myocardial infarction (AMI) diagnosis owing to its superior cardiac specificity for cardiac damage and showing little or no changes in patients with a skeletal muscle disease or trauma. Herein, a new signal amplification surface-enhanced Raman scattering (SERS) platform was developed for recognition and detection of cTnI by using gold nanoparticles (AuNPs), graphene oxide (GO) and magnetic beads (MB). Here, antibody/Raman reporter labeled AuNP-functionalized GO were employed as both SERS nanotags and signal amplification carriers. Monoclonal antibody modified MB were applied as the capture probe and separation agents. In the presence of cTnI, sandwich type immunocomplexes, "capture probe/target/SERS nanotags", were formed through antibody-antigen-antibody interactions. Due to the strong SERS enhancement ability of the designed GO/AuNP complexes and a high binding chance between cTnI and the GO/AuNP complexes, the proposed SERS-based immunoassay could selectively detect cTnI with a high sensitivity (detection limit of 5 pg mL-1) and a good linearity was obtained in a range of 0.01-1000 ng mL-1. In addition, this method was also successfully applied for detecting cTnI in serum substitute media with a similar linear range. Furthermore, this strategy can be constructed with different kinds of antibodies and Raman reporters, and thus can be easily used for simultaneous detection of multiple biomarkers. Therefore, this proposed SERS-based signal amplification technique shows strong potential for the clinical diagnosis of AMI disease.