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Objective: This study aims to assess the early clinical outcomes of bipolar hemiarthroplasty for treating femoral neck fractures in elderly patients aged 75 and above using the Orthopädische Chirurgie München (OCM) approach. Methods: A retrospective analysis was conducted on a cohort of 95 elderly patients who underwent bipolar hemiarthroplasty for Garden Type III and IV femoral neck fractures between January 2020 and December 2022. The participants were categorized into two groups according to the surgical approach used: the OCM approach and the posterior-lateral approach (PLA). The average follow-up duration was 11.20 ± 2.80 months for the OCM group and 11.12 ± 2.95 months for the PLA group, with both groups ranging from 6 to 18 months. Clinical outcomes assessed included surgical duration, incision length, postoperative hospital stay, time to ambulation, hemoglobin levels, serum creatine kinase (CK) levels, C-reactive protein (CRP) levels, pain (assessed using the Visual Analogue Scale, VAS), and functional recovery (evaluated through Harris hip scores). Additionally, complications such as intraoperative and postoperative fractures, deep vein thrombosis, wound infection, nerve injury, postoperative dislocation, leg length discrepancy, and Trendelenburg gait were monitored. Results: There was no significant difference in the surgical duration between the OCM and PLA groups. However, the OCM group exhibited shorter incision lengths, reduced postoperative hospital stays, and earlier ambulation times compared to the PLA group. Significantly lower intraoperative blood loss, smaller decreases in hemoglobin levels on postoperative days 1 and 3, lesser hidden blood loss, and decreased levels of CK and CRP were observed in the OCM group. Pain levels, measured by VAS scores, were lower, and Harris hip scores, indicating functional recovery, were higher at 2 and 6 weeks postoperatively in the OCM group than in the PLA group. The incidence of complications, such as periprosthetic fractures, intramuscular venous thrombosis, hip dislocations, Trendelenburg gait, and leg length discrepancies, showed no significant differences between the groups. Conclusion: The OCM approach for bipolar hemiarthroplasty in patients aged 75 and above with femoral neck fractures offers significant early clinical benefits over the traditional PLA, including faster recovery, reduced postoperative pain, and enhanced early functional recovery.
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Background: The value of semiquantitative resting myocardial perfusion imaging (MPI) in coronary artery disease (CAD) is limited. At present, quantitative MPI can be performed by a new cadmium zinc tellurium single-photon emission computed tomography (CZT-SPECT) scan. The quantitative index of resting myocardial blood flow (MBF) has received little attention, and its manifestations and clinical value in the presence of unstable coronary blood flow have not been clarified. Purpose: In patients with ST-segment elevation myocardial infarction (STEMI), whether resting MBF can provide additional value of blood flow than semi-quantitative resting MPI is not sure. We also explored the influencing factors of resting MBF. Methods: This was a retrospective clinical study. We included 75 patients with STEMI in the subacute phase who underwent resting MPI and dynamic scans after reperfusion therapy. General patient information, STEMI-related data, MPI, gated MPI (G-MPI), and resting MBF data were collected and recorded. According to the clinically provided culprit vessels, the resting MBF was divided into ischemic MBF and non-ischemic MBF. The paired Wilcoxon signed-rank test was used for resting MBF. The receiver operating characteristic (ROC) curves were used to determine the optimal threshold for ischemia, and multiple linear regression analysis was used to analyze the influencing factors of resting MBF. Results: There was a statistically significant difference between the ischemic MBF and non-ischemic MBF [0.59 (0.47-0.72) vs. 0.76 (0.64-0.93), p < 0.0001]. The ROC curve analysis revealed that resting MBF could identify ischemia to a certain extent, with a cutoff value of 0.5975, area under the curve (AUC) = 0.666, sensitivity = 55.8%, and specificity = 68.7%. Male sex and summed rest score (SRS) were influencing factors for resting MBF. Conclusion: To a certain extent, resting MBF can suggest residual ischemia after reperfusion therapy in patients with STEMI. There was a negative correlation between male sex, SRS, and ischemic MBF. A lower resting MBF may be associated with more severe myocardial ischemia.
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Spinal cord injury (SCI) constitutes a significant clinical challenge, and there is extensive research focused on identifying molecular activities that can facilitate the repair of spinal cord injuries. Mammalian sterile 20-like kinase 2 (MST2), a core component of the Hippo signaling pathway, plays a key role in apoptosis and cell growth. However, its role in neurite outgrowth after spinal cord injury remains unknown. Through comprehensive in vitro and in vivo experiments, we demonstrated that MST2, predominantly expressed in neurons, actively participated in the natural development of the CNS. Post-SCI, MST2 expression significantly increased, indicating its activation and potential role in the early stages of neural recovery. Detailed analyses showed that MST2 knockdown impaired neurite outgrowth and motor function recovery, whereas MST2 overexpression led to the opposite effects, underscoring MST2's neuroprotective role in enhancing neural repair. Further, we elucidated the mechanism underlying MST2's action, revealing its interaction with AKT and positive regulation of AKT activity, a well-established promoter of neurite outgrowth. Notably, MST2's promotion of neurite outgrowth and motor functional recovery was diminished by AKT inhibitors, highlighting the dependency of MST2's neuroprotective effects on AKT signaling. In conclusion, our findings affirmed MST2's pivotal role in fostering neuronal neurite outgrowth and facilitating functional recovery after SCI, mediated through its positive modulation of AKT activity. In conclusion, our findings confirmed MST2's crucial role in neural protection, promoting neurite outgrowth and functional recovery after SCI through positive AKT activity modulation. These results position MST2 as a potential therapeutic target for SCI, offering new insights into strategies for enhancing neuroregeneration and functional restoration.
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Objective: To explore the clinical outcomes of a 3D printing-assisted posterolateral approach for the treatment of ankle fractures involving the posterior malleolus. Methods: A total of 51 patients with ankle fractures involving the posterior malleolus admitted to our hospital from January 2018 to December 2019 were selected. The patients were divided into 3D printing group (28 cases) and control group (23 cases). 3D printing was performed for ankle fractures, followed by printing of a solid model and simulation of the operation on the 3D model. The operation was then performed according to the preoperative plan, including open reduction and internal fixation via the posterolateral approach with the patient in the prone position. Routine x-ray and CT examinations of the ankle joint were performed, and ankle function was evaluated using the American Foot and Ankle Surgery Association (AOFAS) ankle-hindfoot score. Results: All patients underwent x-ray and CT examinations. All fractures healed clinically, without loss of reduction or failure of internal fixation. Good clinical effects were achieved in both groups of patients. The operation time, intraoperative blood loss and intraoperative fluoroscopy frequency in the 3D printing group were significantly less than those in the control group (p < 0.05). There was no significant difference between the two groups in the anatomical reduction rate of fractures or the incidence of surgical complications (p > 0.05). Conclusion: The 3D printing-assisted posterolateral approach is effective in the treatment of ankle fractures involving the posterior malleolus. The approach can be well planned before the operation, is simple to perform, yields good fracture reduction and fixation, and has good prospects for clinical application.
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During spinal cord injury (SCI), the homeostasis of the cellular microenvironment in the injured area is seriously disrupted, which makes it extremely difficult for injured neurons with regenerative ability to repair, emphasizing the importance of restoring the cellular microenvironment at the injury site. Neurons interact closely with other nerve cells in the central nervous system (CNS) and regulate these cells. However, the specific mechanisms by which neurons modulate the cellular microenvironment remain unclear. Exosomes were isolated from the primary neurons, and their effects on astrocytes, microglia, oligodendrocyte progenitor cells (OPCs), neurons, and neural stem cells were investigated by quantifying the expression of related proteins and mRNA. A mouse SCI model was established, and neuron-derived exosomes were injected into the mice by the caudal vein to observe the recovery of motor function in mice and the changes in the nerve cells in the lesion area. Neuron-derived exosomes could reverse the activation of microglia and astrocytes and promote the maturation of OPCs in vivo and in vitro. In addition, neuron-derived exosomes promoted neurite outgrowth of neurons and the differentiation of neural stem cells into neurons. Moreover, our experiments showed that neuron-derived exosomes enhanced motor function recovery and nerve regeneration in mice with SCI. Our findings highlight that neuron-derived exosomes could promote the repair of the injured spinal cord by regulating the cellular microenvironment of neurons and could be a promising treatment for spinal cord injury.
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Exosomas , Traumatismos de la Médula Espinal , Ratones , Animales , Exosomas/metabolismo , Neuronas/metabolismo , Traumatismos de la Médula Espinal/patología , Médula Espinal/patología , Microambiente CelularRESUMEN
BACKGROUND: With improved resolution and sensitivity, the cadmium zinc telluride (CZT) detector measures myocardial blood flow (MBF) and myocardial flow reserve (MFR) via single photon emission computed tomography (SPECT). Recently, many studies have used vasodilator stress to obtain quantitative indexes. However, dobutamine used as a pharmaceutical stress has been rarely used to quantify myocardial perfusion using CZT-SPECT. Our study retrospectively analyzed the blood flow performance of 99mTc-Sestamibi (99mTc -MIBI) CZT-SPECT comparing dobutamine to adenosine. PURPOSE: The study aims to explore whether dobutamine stress can be used for the myocardial perfusion quantitative analysis via CZT-SPECT as well as compare dobutamine MBF and MFR to adenosine. METHODS: It was a retrospective study. A total of 68 patients with suspected or known coronary artery disease (CAD) were consecutively enrolled in this study. Thirty-four patients underwent dobutamine stress 99mTc-MIBI CZT-SPECT. Another thirty-four patients underwent adenosine stress 99mTc-MIBI CZT-SPECT. Patient characteristics, myocardial perfusion imaging (MPI) results, gated-myocardial perfusion imaging (G-MPI) results and quantitative analysis of MBF and MFR were collected. RESULTS: In dobutamine stress group, stress MBF was significantly higher than rest MBF (median [interquartile range], 1.63 [1.46-1.94] vs. 0.89 [0.73-1.06], P < 0.001). In adenosine stress group, similar results were observed (median [interquartile range], 2.01 [1.34-2.20] vs. 0.88 [0.75-1.01], P < 0.001). When comparing the dobutamine and adenosine stress group, global MFR showed significant differences (median [interquartile range], the dobutamine group: 1.88 [1.67-2.38] vs. the adenosine group: 2.19 [1.87-2.64], P = 0.037). CONCLUSION: MBF and MFR can be measured using dobutamine 99mTc -MIBI CZT-SPECT. In small sample single-center study, there was a difference in MFR produced by adenosine and dobutamine within the suspected or the known CAD population.
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Enfermedad de la Arteria Coronaria , Imagen de Perfusión Miocárdica , Humanos , Dobutamina , Estudios Retrospectivos , Cadmio , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Tomografía Computarizada de Emisión de Fotón Único/métodos , Tecnecio Tc 99m Sestamibi , Adenosina , Zinc , Imagen de Perfusión Miocárdica/métodosRESUMEN
Edible bird's nest (EBN) is a popular and expensive food material. The limited supply and great demand result in the use of adulterants. The authenticity concern is raised due to the lack of appropriate quality markers. Herein, this study aims to provide a specific oligosaccharide marker for rapid EBN authentication. Comparing the benzocaine (ABEE)-labeled saccharide profiles of multiple batches of EBN and adulterants indicates seven unique EBN oligosaccharides. The most abundant one, named BNM001, was selected as a marker and characterized to be Neu5Ac (2-3) Gal by MS and NMR spectra. This new oligosaccharide marker enables a rapid authentication of EBN within 10 min. ABEE labelling of this marker further upgraded the accuracy and sensitivity of the LC-qTOF-MS quantitative analysis. The relative marker content was associated with the quality of EBN products. These results suggest a specific and efficient quality marker for rapid authentication of EBN and related products.
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Aves , Oligosacáridos , Animales , Carbohidratos , Alimentos , Espectrometría de MasasRESUMEN
Gut barrier makes a huge research gap between in vivo and in vitro studies of orally bioactive polysaccharides: whether/how they contact the related cells in vivo. A hyperbranched heteroglycan RAP from Radix Astragali, exerting antitumor and immunomodulatory effects in vitro and in vivo, is right an example. Here, we determined first that RAP's antitumor activity is immune-dependent. Being undegraded and non-absorbing, RAP quickly entered Peyer's patches (PPs) in 1 h where it directly targeted follicle dendritic cells and initiated antitumor immune responses. RAP was further delivered to mesenteric lymph node, bone marrow, and tumor. By contrast, the control Dendrobium officinale polysaccharide did not enter PPs. These findings revealed a blood/microbiota-independent and selective lymphatic route for orally administrated RAP to directly contact immune cells and trigger antitumor immune responses. This route bridges the research gap between the in vitro and in vivo studies and might apply to many other bioactive polysaccharides.
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Medicamentos Herbarios Chinos , Ganglios Linfáticos Agregados , Astragalus propinquus , Inmunidad , Polisacáridos/metabolismo , Polisacáridos/farmacologíaRESUMEN
OBJECTIVE: To explore the effect of resveratrol in premature senescence and reveal its anti-premature senescence mechanisms through network pharmacology. METHODS: In this study, the H2O2-induced bone marrow mesenchymal stem cells (BMMSCs) premature senescence model is applied. Cell counting kit-8 assay, ß-galactosidase staining and flow cytometry are conducted to detect the proliferation, senescence and apoptosis of BMMSCs. Bioinformatics analyses are used to screen and validate molecular targets of resveratrol acting on premature senescence. Dual-luciferase reporter assay is conducted to verify the interaction between v-rel avian reticuloendotheliosis viral oncogene homolog A (RELA) and sirtuin 1 (SIRT1). RT-qPCR and western blot are adopted to detect mRNA and protein levels of RELA, SIRT1, senescence-related genes and apoptosis-related genes. RESULTS: First, we proved that resveratrol alleviated the H2O2-induced senescence of BMMSCs. Then, bioinformatics analysis revealed that RELA was the downstream target of resveratrol and SIRT1 was the downstream target of RELA, respectively, involved in premature aging. RELA/SIRT1 may be the potential target of resveratrol for premature senescence. Notably, rescue experiments indicated that resveratrol inhibited premature senescence partially through targeting regulation RELA/SIRT1. CONCLUSION: In our study, we confirm the functional role of the resveratrol-RELA- SIRT1 axis in the progression of premature senescence, which provides a latent target for premature senescence treatment.
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Senescencia Celular/efectos de los fármacos , Resveratrol/farmacología , Sirtuina 1/biosíntesis , Factor de Transcripción ReIA/biosíntesis , Apoptosis/efectos de los fármacos , Células Cultivadas , Senescencia Celular/genética , Humanos , Peróxido de Hidrógeno , Células Madre Mesenquimatosas/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
Cordyceps, one of the most expensive natural health supplements, is popularly used to modulate immune function. However, little is known regarding the underlying mechanism of its immunomodulatory activity. We newly reported a Cordyceps quality marker CCP (Mw 433.778 kDa) which was characterized as a 1,4-α glucan by chemical and spectral analysis and is able to induce significant immune responses of macrophages. Herein, we further investigated the molecular mechanism of CCP's immunomodulatory effects. The results indicate that CCP modulates the TLR4/MyD88/p38 signaling pathway of macrophages, where TLR4 plays a crucial role as verified on TLR4-deficient (TLR4-/-) bone marrow-derived macrophages (BMDMs) and TLR4-/- mice. These findings provide a precise understanding of the molecular mechanism of Cordyceps' immunomodulatory benefits.
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Cordyceps/química , Polisacáridos Fúngicos/farmacología , Factores Inmunológicos/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Biomarcadores , Inmunidad/efectos de los fármacos , Interleucina-6/metabolismo , Macrófagos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mariposas Nocturnas , Factor 88 de Diferenciación Mieloide/metabolismo , Células RAW 264.7 , Receptor Toll-Like 2/deficiencia , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/deficiencia , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Radix Astragali polysaccharide RAP has been reported to play a crucial role in hematopoiesis without a clear mechanism. In this study, RAP's effects to enhance the recovery of cyclophosphamide (Cy)-suppressed bone marrow and blood cells is confirmed in vivo first. Confocal micrographs demonstrated the interesting direct binding of FITC-RAP to hematopoietic stem cells (HSC) in bone marrow. RAP protects both mice and human HSC in terms of cell morphology, proliferation, and apoptosis. RNA-sequencing and shRNA approaches revealed FOS to be a key regulator in RAP's protection. These evidences provide an unreported mechanism that RAP directly protects hematopoietic stem cells from chemotherapy-induced myelosuppression by increasing FOS expression.
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Ciclofosfamida/efectos adversos , Medicamentos Herbarios Chinos/química , Hematopoyesis , Células Madre Hematopoyéticas/citología , Polisacáridos/administración & dosificación , Proteínas Proto-Oncogénicas c-fos/genética , Animales , Astragalus propinquus , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Femenino , Células Madre Hematopoyéticas/química , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Ratones , Polisacáridos/farmacología , Análisis de Secuencia de ARN , Regulación hacia ArribaRESUMEN
PURPOSE: To determine the diagnostic performance of virtual noncalcium (VNCa) dual-energy computed tomography (DECT) in the detection of bone marrow edema (BME) in participants with osteonecrosis of the femoral head (ONFH). METHODS: In this prospective study, 24 consecutive participants (15 men, 9 women; mean age, 44 years, range, 21-72 years) diagnosed with ONFH who underwent DECT and magnetic resonance imaging (MRI) between September 2019 and January 2020 were involved. Two independent readers visually evaluated color-coded VNCa images using a binary classification (0 = normal bone marrow, 1 = BME). MRI served as the reference standard for the presence of BME. Interobserver agreement for the visual evaluation of VNCa DECT images was calculated with κ statistics. We determined computed tomography (CT) numbers on VNCa images and weighted-average CT sets using region-of-interest-based quantitative analysis. The t-test was used to compare the differences of CT values between BME areas and normal bone marrow areas. Receiver operating characteristic (ROC) curve was used to select an optimal CT values of VNCa images for detecting BME. A p value of <0.05 was considered as statistically significant. RESULTS: The sensitivity, specificity, and accuracy of Reader 1 and Reader 2, respectively, in the identification of BME at DECT were 95 % and 89 % (18 and 17 of 19), 96 % and 96 % (25 and 25 of 26), and 93 % (43 and 42 of 45). Interobserver agreement was excellent (κ = 0.86). The VNCa CT numbers of the BME area and the normal bone marrow area were -28.6 (-17.9--39.4) HU and -97.9 (-91.3--104.4) HU, respectively, with statistical significance (t = -10.6, p < 0.001). The weighted-average CT numbers of the BME area and the normal bone marrow area were 152.4(122.2-182.7) HU and 121.1(103.6-183.6) HU, respectively, with no statistical significance (t = -2.0, p > 0.05). The area under the receiver operating characteristic curve was 0.99 in differentiation of the BME from normal bone marrow. A cut-off value of -57.2 HU yielded overall sensitivity, specificity, and accuracy, respectively, of 95 % (18 of 19), 100 % (26 of 26), and 98 % (44 of 45) detection of BME in participants with ONFH. CONCLUSION: Visual and quantitative analyses of VNCa images shows excellent diagnostic performance for assessing BME in participants with ONFH.
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Médula Ósea , Osteonecrosis , Adulto , Anciano , Médula Ósea/diagnóstico por imagen , Edema/diagnóstico por imagen , Femenino , Cabeza Femoral , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Polysaccharides have broad bioactivities and are major components of water decoction of herb formulae. However, the quality control of polysaccharides remains a challenge. Oligosaccharide-fragment approach has been considered in elucidating chemical structures of polysaccharides, but never been used for quantitation. Using reference chemicals and a real sample Danggui Buxue Tang (DBT) in this study, an oligosaccharide-marker approach was established to quantify specific polysaccharides. Firstly, linear relationships between parent polysaccharides and hydrolysis-produced daughter oligosaccharides were verified using reference polysaccharides. Then in case of DBT, two fluorescence-labeled oligosaccharides with high specificity to individual parent polysaccharides were selected as markers. They were easily isolated and identified. Their potential in quantification of parent polysaccharides were satisfactorily validated in terms of linearity (r≥0.99), repeatability (RSDâ¯≤â¯8.4 %), and spike recovery (≥80 %). This method could be a promising approach for quality assessment of polysaccharides in herbal formulae.
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Química Farmacéutica/métodos , Medicamentos Herbarios Chinos/análisis , Oligosacáridos/análisis , Control de Calidad , Química Farmacéutica/normas , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , Medicamentos Herbarios Chinos/química , Estándares de Referencia , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masa por Ionización de Electrospray/normas , Espectrometría de Masas en Tándem/métodos , Espectrometría de Masas en Tándem/normasRESUMEN
Polysaccharides from functional foods have been proved to have diverse bioactivities, but little is known about what exactly happens to these polysaccharides after oral administration and even less about the underlying mechanism of action. Taking the marker polysaccharide (DOP) of Dendrobium officinale as an example, this study aims to demonstrate the dynamic distribution and degradation of orally dosed DOP in mice and in vitro using near-infrared fluorescence imaging and a kind of chromatographic analysis. The results indicate that (1) neither DOP nor fluorescence-labeled DOP (FDOP) was absorbed, (2) both DOP and FDOP were undigested and were quickly degraded to short-chain fatty acids in the large intestine, (3) DOP modulated gut microbiota, which could be associated with DOP's suppression of 4T1 tumor growth in mice. All of these findings suggest that some (maybe not all) bioactive polysaccharides share a common destiny: indigestible and nonabsorbing, ends in modulating bioactivities-associated gut microbiota.
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Dendrobium/metabolismo , Microbioma Gastrointestinal , Extractos Vegetales/metabolismo , Polisacáridos/metabolismo , Animales , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Dendrobium/química , Femenino , Alimentos Funcionales/análisis , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/química , Polisacáridos/químicaRESUMEN
Glucuronidation is a major process of drug metabolism and elimination that generally governs drug efficacy and toxicity. Publications have demonstrated that efflux transporters control intracellular glucuronidation metabolism. However, it is still unclear whether and how efflux transporters interact with UDP-glucuronosyltransferases (UGTs) in subcellular organelles. In this study, kaempferol, a model fluorescent flavonoid, was used to investigate the interplay of glucuronidation with transport at the subcellular level. Human recombinant UGTs and microsomes were utilized to characterize the in vitro glucuronidation kinetics of kaempferol. The inhibition of UGTs and efflux transporters on the subcellular disposition of kaempferol were determined visually and quantitatively in Caco-2/TC7 cells. The knockout of transporters on the subcellular accumulation of kaempferol in liver and intestine were evaluated visually. ROS and Nrf2 were assayed to evaluate the pharmacological activities of kaempferol. The results showed that UGT1A9 is the primary enzyme responsible for kaempferol glucuronidation. Visual and quantitative data showed that the UGT1A9 inhibitor carvacrol caused a significant rise in subcellular aglycone and reduction in subcellular glucuronides of kaempferol. The inhibition and knockout of transporters, such as P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), and multidrug resistance-associated proteins (MRPs), exhibited a marked increase in subcellular kaempferol and decrease in its subcellular glucuronides. Correspondingly, inhibition of UGT1A9 and transporters led to increased kaempferol and, consequently, a significantly enhanced ROS scavenging efficiency and nuclear translocation of Nrf2. In conclusion, the interplay of efflux transporters (P-gp, BCRP, and MRPs) and UGTs govern the subcellular exposure and corresponding pharmacological activity of kaempferol.
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Glucurónidos/metabolismo , Glucuronosiltransferasa/metabolismo , Quempferoles/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Animales , Células CACO-2 , Cimenos , Glucuronosiltransferasa/antagonistas & inhibidores , Humanos , Intestinos/citología , Proteínas de Transporte de Membrana/genética , Ratones Noqueados , Microsomas/metabolismo , Monoterpenos/farmacología , Factor 2 Relacionado con NF-E2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , UDP Glucuronosiltransferasa 1A9RESUMEN
Lung adenocarcinoma (LAC), predominant subclassfication of lung cancer, leads high incidence and mortality annually worldwide. During the premalignant transition from lung adenomas to LAC, cellular senescence is regard as a critical physiological barrier against tumor progression. Nevertheless, the role of senescence in tumorigenesis is controversial and few senescence inducers are extensively determined. In this study, we used two classical cell lines A549 and H1299 and two NSCLC xenograft models on Balb/c-nude mice to reveal the pro-senescence effects of shikonin and the corresponding underlying mechanism in LAC. Shikonin, a pure compound isolated from the herbal medicine Lithospermum erythrorhizon, remarkably stimulated cellular senescence including increased SAHF formation, enlarged cellular morphology, and induced SA-ß-Gal positive staining. Further mechanism study revealed that the pro-senescence effect of shikonin was dependent on the increased intercellular ROS generation, which subsequently triggered DNA damage-p53/p21waf axis without activating oncogenes such as Ras and MEK-1. Meanwhile, Kdm2b, an H3K36me2-specific demethylase effectively suppressed ROS generation, was also notably suppressed by shikonin treatment. Moreover, shikonin at 10 mg/kg significantly inhibited tumor weights by 55.84% and 50.98% in A549 and H1299 xenograft model, respectively (P < 0.05) through activating cellular senescence. Our study suggested that shikonin, a ROS-dependent senescence inducer, could serve as a promising agent for further lung cancer treatment.
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Lung cancer is a leading cause of cancer-related deaths worldwide. NOTCH3 signaling is mainly expressed in non-small cell lung carcinoma (NSCLC), and has been proposed as a therapeutic target of NSCLC. While, few agents for preventing or treating NSCLC via targeting NOTCH3 signaling are used in modern clinical practice. Evodiamine (EVO), an alkaloid derived from Euodiae Fructus, possesses low toxicity and has long been shown to exert anti-lung cancer activity. However, the underlying anti-lung cancer mechanisms of EVO are not yet fully understood. In this study, we explored the involvement of NOTCH3 signaling in the anti-lung cancer effects of EVO. Urethane-induced lung cancer mouse model and two NSCLC cell models, A549 and H1299, were used to evaluate the in vivo and in vitro anti-lung cancer action of EVO. A DNA methyltransferase inhibitor was employed to investigate the role of NOTCH3 signaling in the anti-lung cancer effects of EVO. Results showed that EVO potently reduced tumor size and tumor numbers in mice, and inhibited NOTCH3 in the tumors. EVO also dramatically reduced cell viability, induced G2/M cell cycle arrest, inhibited cell migration and reduced stemness in cultured NSCLC cells. Mechanistic studies showed that EVO potently inhibited NOTCH3 signaling by activation of DNMTs-induced NOTCH3 methylation. Importantly, inhibition of NOTCH3 methylation in NSCLC cells diminished EVO's anti-NSCLC effects. Collectively, EVO, a novel NOTCH3 methylation stimulator, exerted potent anti-lung cancer effects partially by inhibiting NOTCH3 signaling. These findings provide new insight into the EVO's anti-NSCLC action, and suggest a potential role of EVO in lung cancer prevention and treatment.
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ETHNOPHARMACOLOGICAL RELEVANCE: Camptosorus sibiricus Rupr (CSR) is a widely used herbal medicine with antivasculitis, antitrauma, and antitumor effects. However, the effect of CSR aqueous extract on B[a]P-initiated tumorigenesis and the underlying mechanism remain unclear. Moreover, the compounds in CSR aqueous extract need to be identified and structurally characterized. AIM OF THE STUDY: We aim to investigate the chemopreventive effect of CSR and the underlying molecular mechanism. MATERIALS AND METHODS: A B[a]P-stimulated normal cell model (BEAS.2B) and lung adenocarcinoma animal model were established on A/J mice. In B[a]P-treated BEAS.2B cells, the protective effects of CSR aqueous extract on B[a]P-induced DNA damage and ROS production were evaluated through flow cytometry, Western blot, real-time quantitative PCR, single-cell gel electrophoresis, and immunofluorescence. Moreover, a model of B[a]P-initiated lung adenocarcinoma was established on A/J mice to determine the chemopreventive effect of CSR in vivo. The underlying mechanism was analyzed via immunohistochemistry and microscopy. Furthermore, the new compounds in CSR aqueous extract were isolated and structurally characterized using IR, HR-ESI-MS, and 1D and 2D NMR spectroscopy. RESULTS: CSR effectively suppressed ROS production by re-activating Nrf2-mediated reductases HO-1 and NQO-1. Simultaneously, CSR attenuated the DNA damage of BEAS.2B cells in the presence of B[a]P. Moreover, CSR at 1.5 and 3â¯g/kg significantly suppressed tumorigenesis with tumor inhibition ratios of 36.65% and 65.80%, respectively. The tumor volume, tumor size, and multiplicity of B[a]P-induced lung adenocarcinoma were effectively decreased by CSR in vivo. After extracting and identifying the compounds in CSR aqueous extract, three new triterpene saponins were isolated and characterized structurally. CONCLUSIONS: CSR aqueous extract prevents lung tumorigenesis by exerting dual effects against ROS and DNA damage, suggesting that CSR is a novel and effective agent for B[a]P-induced carcinogenesis. Moreover, by isolating and structurally characterizing three new triterpene saponins, our study further standardized the quality of CSR aqueous extract, which could widen CSR clinical applications.
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Adenocarcinoma/prevención & control , Anticarcinógenos/farmacología , Helechos/química , Neoplasias Pulmonares/prevención & control , Extractos Vegetales/farmacología , Adenocarcinoma del Pulmón , Animales , Anticarcinógenos/aislamiento & purificación , Benzo(a)pireno/toxicidad , Western Blotting , Daño del ADN/efectos de los fármacos , Citometría de Flujo , Humanos , Ratones , Extractos Vegetales/química , Especies Reactivas de Oxígeno/metabolismo , Saponinas/aislamiento & purificación , Triterpenos/química , Triterpenos/aislamiento & purificaciónRESUMEN
DACT2, a tumor suppressor gene in various tumors, is frequently down-regulated via hypermethylation. We found DACT2 gene expressions were dramatically silenced (Pâ¯=â¯0.002, nâ¯=â¯8) in our clinical colorectal cancer (CRC) tissues, and TCGA data revealed DACT2 hypermethylation correlated to CRC poor prognosis (Pâ¯=â¯0.0129, HRâ¯=â¯0.2153, nâ¯=â¯248). Thus, by screening twelve nutritional compounds, we aimed to find out an effective DACT2 epigenetic stimulator to determine whether DACT2 epigenetic restoration could reverse CRC tumorigenesis. We found that kaempferol significantly increased DACT2 expressions up to 3.47-fold in three CRC cells (HCT116, HT29, and YB5). Furthermore, kaempferol remarkably decreased DACT2 methylation (range: 19.58%-67.00%, Pâ¯<â¯0.01), while increased unmethylated DACT2 by 13.72-fold (Pâ¯<â¯0.01) via directly binding to DNA methyltransferases DNMT1. By epigenetic reactivating DACT2 transcription, kaempferol notably inhibited nuclear ß-catenin expression to inactivate Wnt/ß-catenin pathway, which consequently restricted CRC cells proliferation and migration. Moreover, in AOM/DSS-induced CRC tumorigenesis, kaempferol-demethylated DACT2 effectively decreased tumor load (range: 50.00%-73.52%, Pâ¯<â¯0.05). By determining the chemopreventive and chemotherapeutic efficacy of a novel DACT2 demethylating stimulator, we demonstrated that DACT2 epigenetic restoration could successfully slow down and reverse CRC tumorigenesis.
Asunto(s)
Proteínas Portadoras/genética , Neoplasias Colorrectales/genética , Proteínas de Neoplasias/genética , Proteínas Adaptadoras Transductoras de Señales , Animales , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/prevención & control , Epigénesis Genética , Humanos , Quempferoles/farmacología , Masculino , Ratones Endogámicos C57BLRESUMEN
Connexin 43 (Cx43), a vital gap junction protein in tumor microenvironment (TME), is a novel molecular target for melanoma chemotherapeutics due to its tumor suppressive function. Dioscin, an herbal steroidal saponin, exerts anti-tumor effects while the underlying mechanism is unclear. Using WB, FACS, and immunofluorescence methodologies, we found dioscin significantly activated the transcription and translation of Cx43 via the retinoid acid signaling pathway and simultaneously enhanced the transporting function of Cx43. Through stimulating Cx43, dioscin remarkably suppressed the migratory and invasive capacities of B16 cells, and notably decreased pluripotency markers of cancer stem cells and epithelial-to-mesenchymal transition in B16 cells and animal tumor tissues. Conversely, dioscin improved the secretion of pro-inflammatory cytokines (IL-6, TNFα, and IL-1ß), and the phagocytic capacity of tumor-associated macrophages by increasing M2-to-M1 phenotype transition. More strikingly, even in Cx43 functional deficient B16 and RAW264.7 cells, dioscin still dramatically reversed the aggravated tumor malignancy and reduced macrophage phagocytic activity. Two classical metastasis animal models were utilized in vivo and results showed that dioscin showed significant anti-metastatic effects, which is closely related to the expression of Cx43 either in in situ tumor or metastatic lung nodes. In conclusion, dioscin targets Cx43 to suppress the tumor cell malignancy and activate macrophage sensitivity, thereby targeting melanoma microenvironment.