RESUMEN
OBJECTIVE: Our research aimed to investigate the therapeutic effects of Tubastatin-A, a glucocorticoid receptor (GR) mitochondrial translocation inhibitor, and mitoquinone (MitoQ), an antioxidant, on attenuating dexamethasone (DEX)-induced macrophage apoptosis. METHODS: We treated RAW264.7 macrophages with different combinations of DEX and either Tubastatin-A or MitoQ. Parameters such as mitochondrial GR translocation, mitochondrial reactive oxygen species levels, mitochondrial membrane potential, mitochondrial permeability transition pore opening, cytochrome C efflux to the cytosol, and apoptosis were subsequently evaluated in the different treatment groups via qRT-PCR, western blotting, and immunofluorescence assays. RESULTS: DEX intervention increased the translocation of GRs into the mitochondria, while reducing the expression of the mitochondrial gene MT-CO1 and the activity of mitochondrial respiratory chain complex IV in macrophages. In addition, DEX administration increased mtROS levels, mitochondrial permeability transition pore opening, and mitochondrial cytochrome C release in macrophages, which promoted their apoptosis. We found that Tubastatin-A inhibited mitochondrial GR translocation and reversed the DEX-induced increase in GR levels within the mitochondria. Furthermore, Tubastatin-A mitigated various mitochondrial changes induced by DEX, including reducing the efflux of mitochondrial cytochrome C and inhibiting macrophage apoptosis. Similarly, MitoQ exerted its effects on macrophage apoptosis by reducing mtROS levels through the mitochondrial pathway. CONCLUSIONS: The DEX-mediated translocation of GR into mitochondria disrupts the mitochondrial function of macrophages, which induces their apoptosis. By inhibiting mitochondrial translocation of GR and reducing mtROS levels, Tubastatin-A and MitoQ can effectively attenuate macrophage apoptosis, which has clinical implications for reducing the notable side effects associated with glucocorticoid use.
Asunto(s)
Apoptosis , Dexametasona , Glucocorticoides , Macrófagos , Mitocondrias , Receptores de Glucocorticoides , Receptores de Glucocorticoides/metabolismo , Animales , Ratones , Apoptosis/efectos de los fármacos , Células RAW 264.7 , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Dexametasona/farmacología , Glucocorticoides/farmacología , Transporte de Proteínas/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Ubiquinona/farmacología , Ubiquinona/análogos & derivados , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Compuestos OrganofosforadosRESUMEN
The PI3K/AKT/mTOR signaling pathway is one of the most common abnormal activation pathways in tumor cells, and has associated with multiple functions such as tumor cell growth, proliferation, migration, invasion, and tumor angiogenesis. Here, a series of 3-amino-1H-indazole derivatives were synthesized, and their antiproliferative activities against HT-29, MCF-7, A-549, HepG2 and HGC-27 cells were evaluated. Among them, W24 exhibited the broad-spectrum antiproliferative activity against four cancer cells with IC50 values of 0.43-3.88 µM. Mechanism studies revealed that W24 inhibited proliferation by affecting the DNA synthesis, induced G2/M cell cycle arrest and apoptosis by regulating Cyclin B1, BAD and Bcl-xL, meanwhile induced the change of intracellular ROS and mitochondrial membrane potential in HGC-27 cells. Moreover, W24 inhibited the migration and invasion of HGC-27 cells by decreasing EMT pathway related proteins and reducing the mRNA expression levels of Snail, Slug and HIF-1α. Furthermore, W24 displayed low tissue toxicity profile and good pharmacokinetic properties in vivo. Therefore, 3-amino-1H-indazole derivatives might serve as a new scaffold for the development of PI3K/AKT/mTOR inhibitor and anti-gastric cancer reagent.
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Indazoles , Neoplasias , Humanos , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Neoplasias/tratamiento farmacológico , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo , Indazoles/química , Indazoles/farmacologíaRESUMEN
A novel actinomycete strain, designated H11425T, was isolated from a sediment sample collected from Baihua Lake, Guizhou Province, PR China, and a polyphasic approach was employed to determine its taxonomic position. 16S rRNA gene sequence comparisons showed that strain H11425T is most closely related to Pseudonocardia sulfidoxydans JCM 10411T (97.9%) and Pseudonocardia kunmingensis JCM 32122T (97.8%). Both of phylogenetic analysis based on 16S rRNA gene sequence and phylogenomic analysis based on whole-genome sequence showed that strain H11425T formed a separate clade within the genus Pseudonocardia. The draft genome had a length of 8,059,576 bp with a G + C content of 74.5%. The average nucleotide identity, average amino acid identity, and digital DNA-DNA hybridization values between strain H11425T and its closely related Pseudonocardia species were 76.8-79.0%, 64.8-69.9% and 21.7-23.3%, respectively, which were significantly lower than the widely accepted species-defined threshold. Strain H11425T contained meso-diaminopimelic acid, arabinose, galactose, glucose and ribose in its whole-cell hydrolysates. Mycolic acids were absent. The menaquinone was identifed as MK-8(H4). The phospholipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylcholine, an unknown phospholipid and four unidentified aminophospholipids. The major fatty acids were iso-C16:0, iso-C14:0, iso H-C16:1 and iso-C16:0 2OH. On the basis of the taxonomic evidence, strain H11425T represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia lacus sp. nov. is proposed. The type strain is H11425T (= JCM 34851T = CICC 25118T).
Asunto(s)
Actinobacteria , Actinomycetales , Actinobacteria/genética , Pseudonocardia , Fosfatidiletanolaminas , Lagos , Filogenia , ARN Ribosómico 16S/genética , Fosfolípidos , ADNRESUMEN
OBJECTIVE: To conduct a systematic review and meta-analysis of randomized controlled trials (RCTs) on the effects of orthokeratology for slowing myopia progression in children. METHODS: We performed a specific search on PubMed, Embase, Cochrane Library, Clinical Trials, CNKI, SinoMed, and Wanfang Data for RCTs conducted up to October 1, 2022. We pooled the weighted mean difference (WMD) between the orthokeratology and control groups for axial length (AL) elongation and the odds ratio (OR) for rates of adverse events and dropout. RESULTS: Seven RCTs involving 655 eyes were included. There were significant differences in the effects of orthokeratology versus control in slowing AL elongation with WMD of -0.11 mm (95% confidence interval (CI), -0.13 to -0.08; P <0.01) at 6 months, -0.16 mm (95% CI, -0.18 to -0.13; P <0.01) at 12 months, -0.23 mm (95% CI, -0.29 to -0.18; P <0.01) at 18 months, and -0.28 mm (95% CI, -0.38 to -0.19; P <0.01) at 24 months, respectively. Myopia control rate declined, with 64%, 53%, 50%, and 47% recorded for 6, 12, 18, and 24 months, respectively. There was no statistical significance for adverse events between orthokeratology and control groups (OR=2.63, 95% CI, 0.72-9.61; P =0.11). CONCLUSION: Orthokeratology can effectively slow myopia progression in children, and the efficacy of myopia control decreases with time.
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Miopía , Procedimientos de Ortoqueratología , Niño , Humanos , Ensayos Clínicos Controlados Aleatorios como Asunto , Miopía/terapia , Longitud Axial del Ojo , Oportunidad Relativa , Refracción OcularRESUMEN
A new series of N-(3,4,5-trimethoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-amine derivatives as tubulin polymerization inhibitors were synthesized, and evaluated for the anti-proliferative activities. A structure-activity relationship study revealed that the free amino moiety of 1H-pyrazolo[3,4-b]pyridin-3-amine played an essential role in the anti-proliferative activities. Especially, compound 15c displayed the strongest anti-proliferation against MCF-7 cells with IC50 value of 0.067 ± 0.003 µM, and high selectivity over the normal human embryonic lung WI-38 cells with IC50 value of 23.41 ± 1.53 µM. Further mechanistic studies revealed that 15c showed strong anti-tubulin polymerization activity, changed the morphology of tubulin, and arrested the cell cycle at the G2/M transition in MCF-7 cells. Molecular docking analysis suggested that 15c well occupied the colchicine-binding pocket of tubulin. Additionally, 15c demonstrated anti-angiogenic activities with blocking the migration, invasion and tube formation, disrupting the newly formed tube, and regulating both MMP-9 and TIMP-1 in HUVEC cells. In summary, our results highlight that compound 15c is a potential antitumor compound that are worthy of further development.
Asunto(s)
Aminas/farmacología , Antineoplásicos/farmacología , Pirazoles/farmacología , Piridinas/farmacología , Moduladores de Tubulina/farmacología , Tubulina (Proteína)/metabolismo , Aminas/síntesis química , Aminas/química , Animales , Antineoplásicos/síntesis química , Antineoplásicos/química , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Polimerizacion/efectos de los fármacos , Pirazoles/síntesis química , Pirazoles/química , Piridinas/síntesis química , Piridinas/química , Relación Estructura-Actividad , Porcinos , Moduladores de Tubulina/síntesis química , Moduladores de Tubulina/químicaRESUMEN
Drought stress, which is increasing with climate change, is a serious threat to agricultural sustainability worldwide. Seed germination is an essential growth phase that ensures the successful establishment and productivity of soybean, which can lose substantial productivity in soils with water deficits. However, only limited genetic information is available about how germinating soybean seeds may exert drought tolerance. In this study, we examined the germinating seed drought-tolerance phenotypes and genotypes of a panel of 259 released Chinese soybean cultivars panel. Based on 4616 Single-Nucleotide Polymorphisms (SNPs), we conducted a mixed-linear model GWAS that identified a total of 15 SNPs associated with at least one drought-tolerance index. Notably, three of these SNPs were commonly associated with two drought-tolerance indices. Two of these SNPs are positioned upstream of genes, and 11 of them are located in or near regions where QTLs have been previously mapped by linkage analysis, five of which are drought-related. The SNPs detected in this study can both drive hypothesis-driven research to deepen our understanding of genetic basis of soybean drought tolerance at the germination stage and provide useful genetic resources that can facilitate the selection of drought stress traits via genomic-assisted selection.
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Sequías , Regulación del Desarrollo de la Expresión Génica , Estudio de Asociación del Genoma Completo , Germinación , Glycine max/genética , Polimorfismo de Nucleótido Simple , Semillas/genética , Mapeo Cromosómico , Regulación de la Expresión Génica de las Plantas , Genotipo , Desequilibrio de Ligamiento , Fenotipo , Proteínas de Plantas/genética , Sitios de Carácter Cuantitativo , Semillas/crecimiento & desarrollo , Glycine max/crecimiento & desarrollo , Estrés FisiológicoRESUMEN
Serine/threonine protein kinases Aurora A, B, and C play essential roles in cell mitosis and cytokinesis, and a number of Aurora kinase inhibitors have been evaluated in the clinic. Herein we report the synthesis and their antiproliferation of 3,5-disubstituted-2-aminopyrazines as kinases inhibitors. Amongst, 4-((3-amino-6- (3,5-dimethylisoxazol-4-yl)pyrazin-2-yl)oxy)-N-(3-chlorophenyl) benzamide (12Aj) exhibited the strongest antiproliferative activities against U38, HeLa, HepG2 and LoVo cells with IC50 values were 11.5 ± 3.2, 1.34 ± 0.23, 7.30 ± 1.56 and 1.64 ± 0.48 µM, as well as inhibited Aurora A and B with the IC50 values were 90 and 152 nM, respectively. Molecular docking studies indicated that 12Aj appeared to form stable hydrogen bonds with either Aurora A or Aurora B. Furthermore, 12Aj arrested HeLa cell cycle in G2/M phase by regulating protein levels of cyclinB1 and cdc2. In addition, the bioinformatics prediction further revealed that 12Aj possessed good drug likeness using SwissADME. These results suggested that 12Aj was worthy of future development of potent anticancer agents as pan-Aurora kinases.
Asunto(s)
Antineoplásicos/farmacología , Aurora Quinasa A/antagonistas & inhibidores , Aurora Quinasa B/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Pirazinas/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Aurora Quinasa A/metabolismo , Aurora Quinasa B/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Pirazinas/síntesis química , Pirazinas/química , Relación Estructura-ActividadRESUMEN
A series of conjugates of podophyllotoxin and coumarin were prepared using the click reaction, and their cytotoxicities against A549, HepG2, HeLa, and LoVo cells were evaluated. Among them, compound 14e exhibited the strongest cytotoxicities against these cancer cells with IC50 values of 4.9-17.5⯵M. Furthermore, 14e disrupted microtubules and induced cell cycle arrest at G1 phase by regulating P21 and Cyclin D1 in LoVo cells. In addition, 14e bond CT DNA and selectively inhibited Topo IIß over Topo IIα. Molecular docking model showed that 14e appeared to form stable hydrogen bonds with several DNA bases and residue Gln778. Taken together, these conjugates have the potential to be developed as anti-tumor drugs.
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Puntos de Control del Ciclo Celular/efectos de los fármacos , Cumarinas/uso terapéutico , ADN/metabolismo , Podofilotoxina/química , Cumarinas/farmacología , ADN-Topoisomerasas de Tipo II/metabolismo , Humanos , Relación Estructura-ActividadRESUMEN
The Aurora kinases are a family of serine/threonine kinases that interact with components of the mitotic apparatus and serve as potential therapeutic targets in oncology. Herein, we reported a series of 2,4-bisanilinopyrimidines bearing 2,2,6,6-tetramethylpiperidine-N-oxyl with selective inhibition of Aurora A in either enzymatic assays or cellular phenotypic assays, and displaying more potent anti-proliferation compared with that of VX-680. The most potent compound 10a forms better interaction with Aurora A than Aurora B in molecular docking. Mechanistic studies revealed that 10a disrupt the spindle formation, block the cell cycle progression in the G2/M phase and induce apoptosis in HeLa cell. These results suggested that the produced series of compounds are potential anticancer agents for further development as selective Aurora A inhibitors.
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Compuestos de Anilina/farmacología , Óxidos N-Cíclicos/farmacología , Piperidinas/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Pirimidinas/farmacología , Compuestos de Anilina/síntesis química , Compuestos de Anilina/química , Apoptosis/efectos de los fármacos , Aurora Quinasa A/química , Sitios de Unión , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Óxidos N-Cíclicos/síntesis química , Óxidos N-Cíclicos/química , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Humanos , Simulación del Acoplamiento Molecular , Piperidinas/síntesis química , Piperidinas/química , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Pirimidinas/síntesis química , Pirimidinas/química , Huso Acromático/efectos de los fármacosRESUMEN
PURPOSE: To investigate the prevalence of macular abnormalities in Chinese patients affected by uveitis. METHODS: A retrospective study was performed by reviewing the medical records and high-definition spectral domain optical coherence tomography (SD-OCT) scans in a cohort of 413 uveitis eyes to assess the presence of macular abnormalities. RESULTS: Macular abnormalities were observed in 242 (58.6%) of the 413 examined eyes with uveitis. The observed macular abnormalities include diffuse macular edema, cystoid macular edema (CME), and serous retinal detachment. In addition, vitreoretinal abnormalities in the patients with uveitis were evaluated through SD-OCT. The evaluated vitreoretinal abnormalities include epiretinal membrane, tractional CME, macular adhesions, macular hole, and choroidal neovascularization. The most frequent abnormality in the uveitis patients was CME, observed in 105 eyes (25.4%), and followed by epiretinal membrane, observed in 52 eyes (12.6%). Moreover, the percentage of uveitis patients with vitreoretinal abnormalities (23.1%) was significantly high. CONCLUSIONS: Macular abnormalities were more frequently observed in uveitis compared with those in the general population. Multiple patterns of macular abnormalities indicate lesions of multilayer of cells. Therefore, when following up uveitis patients, it is highly recommended to screen with SD-OCT to evaluate the macular alterations of those patients. Careful observation of the macular abnormalities using SD-OCT may help to identify patients who are at risk for visual loss secondary to foveal changes and provide preventive treatment.
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Pueblo Asiatico , Mácula Lútea/patología , Enfermedades de la Retina/epidemiología , Uveítis/epidemiología , Adolescente , Adulto , Anciano , China/epidemiología , Femenino , Angiografía con Fluoresceína , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Enfermedades de la Retina/diagnóstico , Estudios Retrospectivos , Tomografía de Coherencia Óptica/métodos , Uveítis/diagnóstico , Agudeza Visual/fisiologíaRESUMEN
PURPOSE: To conduct a meta-analysis on the effects of orthokeratology in slowing myopia progression. METHODS: A literature search was performed in PubMed, Embase, and the Cochrane Library. Methodological quality of the literature was evaluated according to the Jadad score. The statistical analysis was carried out using RevMan 5.2.6 software. RESULTS: The present meta-analysis included seven studies (two randomized controlled trials and five nonrandomized controlled trials) with 435 subjects (orthokeratology group, 218; control group, 217) aged 6 to 16 years. The follow-up time was 2 years for the seven studies. The weighted mean difference was -0.26 mm (95% confidence interval, -0.31 to -0.21; p < 0.001) for axial length elongation based on data from seven studies and -0.18 mm (95% confidence interval, -0.33 to -0.03; p = 0.02) for vitreous chamber depth elongation based on data from two studies. CONCLUSIONS: Our results suggest that orthokeratology may slow myopia progression in children. Further large-scale studies are needed to substantiate the current result and to investigate the long-term effects of orthokeratology in myopia control.
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Miopía/prevención & control , Procedimientos de Ortoqueratología , Adolescente , Niño , Ensayos Clínicos Controlados como Asunto , HumanosRESUMEN
PURPOSE: Angiogenesis is an important mediator in tumor progression. Vascular endothelial growth factor (VEGF) is one of the major cytokines that can influence angiogenesis. However, the potential mechanism of tumor growth inhibition through anti-VEGF agents is still unclear. This study was performed to examine whether ranibizumab could inhibit malignant melanoma growth in vitro and to determine the safety of ranibizumab on human adult retinal pigment epithelium cell line (ARPE-19 cells). METHODS: Malignant melanoma cells obtained from a clinic were cultured in vitro. VEGF concentrations secreted by malignant melanoma cells and the ARPE-19 cells were examined by enzyme-linked immunosorbent assay (ELISA). The two kinds of cells were both treated with VEGF and its antagonist, ranibizumab. The dynamic changes of the two types of cells were monitored by real-time cell electronic sensing (RT-CES) assay. The effect of ranibizumab on both types of cells was verified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT) assay. The expression of VEGF receptor 1 (VEGFR1) RNA in uveal melanoma was further investigated through the PCR technique. RESULTS: The levels of VEGF secreted by malignant melanoma cells were much higher than those of ARPE-19 cells, and were markedly decreased in the action of 0.1 mg/ml ranibizumab. However, there was no obvious reduction of VEGF in the presence of ranibizumab for ARPE-19 (p>0.05). Meanwhile, RT-CES showed that the viability of malignant melanoma cells increased greatly in the presence of VEGF. When VEGF was 20 ng/ml, viability of the malignant melanoma cells increased by 40% compared with the negative control. There was no evident effect on proliferation of ARPE-19 (p>0.05). Furthermore, the growth of malignant melanoma cells was obviously inhibited after ranibizumab intervention. When ranibizumab was administered at 0.25 mg/ml, the survival rate of the malignant melanoma cells decreased to 57.5%. Nevertheless, low-dose exposure to ranibizumab had only a slight effect on the growth of ARPE-19, and PCR result demonstrated that VEGFR1 plays a role in this tumor tissue rather than VEGFR2. CONCLUSIONS: Ranibizumab can selectively inhibit malignant melanoma cell proliferation by decreasing the expression of VEGF; the possible mechanism of the inhibitory effect may involve VEGFR1 antagonism.
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Anticuerpos Monoclonales Humanizados/uso terapéutico , Melanoma/metabolismo , Melanoma/patología , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismo , Adulto , Anticuerpos Monoclonales Humanizados/farmacología , Muerte Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/patología , Sistemas de Computación , Humanos , Inmunohistoquímica , Melanoma/tratamiento farmacológico , Ranibizumab , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/patología , Neoplasias de la Úvea/tratamiento farmacológico , Neoplasias de la Úvea/patología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: The identification of the biomarkers of patients with acute anterior uveitis (AAU) may allow for a less invasive and more accurate diagnosis, as well as serving as a predictor in AAU progression and treatment response. The aim of this study was to identify the potential biomarkers and the metabolic pathways from plasma in patients with AAU. METHODS: Both plasma metabolic biomarkers and metabolic pathways in the AAU patients versus healthy volunteers were investigated using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and a metabonomics approach. The principal component analysis (PCA) was used to separate AAU patients from healthy volunteers as well as to identify the different biomarkers between the two groups. Metabolic compounds were matched to the KEGG, METLIN, and HMDB databases, and metabolic pathways associated with AAU were identified. RESULTS: The PCA for UPLC-MS data shows that the metabolites in AAU patients were significantly different from those of healthy volunteers. Of the 4,396 total features detected by UPLC-MS, 102 features were significantly different between AAU patients and healthy volunteers according to the variable importance plot (VIP) values (greater than two) of partial least squares discriminate analysis (PLS-DA). Thirty-three metabolic compounds were identified and were considered as potential biomarkers. Meanwhile, ten metabolic pathways were found that were related to the AAU according to the identified biomarkers. CONCLUSIONS: These data suggest that metabolomics study can identify potential metabolites that differ between AAU patients and healthy volunteers. Based on the PCA, PLS-DA, several potential metabolic biomarkers and pathways in AAU patients were found and identified. In addition, the UPLC-MS technique combined with metabonomics could be a suitable systematic biology tool in research in clinical problems in ophthalmology, and can provide further insight into the pathophysiology of AAU.
Asunto(s)
Biomarcadores/sangre , Metabolómica , Uveítis Anterior/sangre , Enfermedad Aguda , Adulto , Cromatografía Líquida de Alta Presión , Femenino , Voluntarios Sanos , Humanos , Masculino , Espectrometría de Masas , Metaboloma , Análisis de Componente PrincipalRESUMEN
PURPOSE: Spectral-domain optical coherence tomography (SD-OCT) is a noninvasive technique that can provide high-resolution images of macular morphology. The purpose of this study was to examine the pathological mechanism of uveitis and compare the changes in the macula of uveitis patients and the histopathological features of experimentally induced uveitis in mice. METHODS: Macular OCT was performed on 78 eyes of 51 patients of the Eye Hospital of Shandong University of Traditional Chinese Medicine, China, with apparent uveitis changes. C57BL/6 mice were injected with interphotoreceptor retinoid-binding protein (IRBP)-specific T cells from naïve mice immunized with complete Freund adjuvant IRBP(1-20) to induce uveitis. The disease was monitored by indirect fundoscopy. The eyes of the mice with experimental autoimmune uveitis (EAU) were enucleated 18 days after injection and classified according to pathological characteristics. RESULTS: The characteristics of uveitis were classified into six categories. Macular edema was detected in 48 eyes (61.5%); macular epiretinal membrane in 22 eyes (28.2%); choroidal neovascularization and macular lamellar holes in 4 eyes (5.1%), respectively; macular atrophy in 10 eyes (12.8%); and serous neuroepithelium detachment in 22 eyes (28.2%). As in human patients, pathological examinations of mouse EAU showed inflammation, folds, and atrophy of the outer part of the neuroretina, choroidal neovascularization with hemorrhagic retinal detachment, serous neuroepithelium detachment, and epiretinal membrane formation. CONCLUSIONS: Macular OCT of uveitis patients can display different morphological characteristics. Mouse EAU can simulate human uveitis. The comparative analysis of macular OCT in human uveitis and transfer EAU histopathology changes could provide important information on the pathogenesis of human uveitis.
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Modelos Animales de Enfermedad , Mácula Lútea/patología , Tomografía de Coherencia Óptica/métodos , Uveítis/diagnóstico , Traslado Adoptivo , Adulto , Animales , Neovascularización Coroidal/diagnóstico , Membrana Epirretinal/diagnóstico , Proteínas del Ojo , Femenino , Angiografía con Fluoresceína , Humanos , Edema Macular/diagnóstico , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Oftalmoscopía , Desprendimiento de Retina/diagnóstico , Perforaciones de la Retina/diagnóstico , Proteínas de Unión al Retinol , Linfocitos T/inmunología , Uveítis/inmunologíaRESUMEN
Drought is a natural disaster that profoundly impacts on global agricultural production, significantly reduces crop yields, and thereby poses a severe threat to worldwide food security. Addressing the challenge of effectively improving crop drought resistance (DR) to mitigate yield loss under drought conditions is a global issue. An optimal root system architecture (RSA) plays a pivotal role in enhancing crops' capacity to efficiently uptake water and nutrients, which consequently strengthens their resilience against environmental stresses. In this review, we discuss the compositions and roles of crop RSA and summarize the most recent developments in augmenting drought tolerance in crops by manipulating RSA-related genes. Based on the current research, we propose the potential optimal RSA configuration that could be helpful in enhancing crop DR. Lastly, we discussed the existing challenges and future directions for breeding crops with enhanced DR capabilities through genetic improvements targeting RSA.
RESUMEN
OBJECTIVE: To assess the effect of luteinized unruptured follicles (LUF) on frozen-thawed embryo transfer cycles performed in natural cycles (FET-NC). METHODS: Retrospective cohort study, held in a university hospital with 3415 cycles for frozen-thawed embryo transfer, performed between June 2019 and September 2022. Using propensity score matching, 242 patients with a diagnosis of LUF (LUF group) were matched with 484 ovulated patients (ovulation group). Stratified by the type of embryo transferred, the LUF group included 168 blastocyst transfer patients (blastocyst group) and 74 cleavage-stage embryo transfer patients (cleavage-embryo group). The ovulation group included 324 patients with blastocyst transfer (blastocyst group) and 160 patients with transferred cleavage-stage embryos. Clinical pregnancy rate was retrospectively analyzed between the LUF and ovulation groups, as well as between each subgroup. RESULTS: After using propensity score matching, the general characteristics of the LUF and ovulation groups were similar. The implantation and clinical pregnancy rates in the LUF group were not significantly different from those in the ovulation group (44.98 % vs. 45.29 %, p = 0.93; 53.72 % vs. 52.48 %, p = 0.75). The implantation and pregnancy rates of transferred cleavage-stage embryos in the LUF group were also not significantly different from those in the ovulation group (32.39 % vs. 36.40 %, p = 0.42; 47.30 % vs. 47.50 %, p = 0.98). The implantation and pregnancy rates of transferred blastocysts in the LUF group were also not significantly different from those in the ovulation group (53.11 % vs. 52.03 %, p = 0.82; 56.55 % vs. 54.94 %, p = 0.73). There was also no significant difference in the miscarriage rate between the groups. CONCLUSION: In the natural cycle, LUF does not affect the clinical pregnancy outcomes of FET. If adequate luteal support is given, the clinical pregnancy outcomes were similar between the LUF group and ovulation group.
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Criopreservación , Transferencia de Embrión , Embarazo , Femenino , Humanos , Estudios Retrospectivos , Índice de Embarazo , Compuestos OrgánicosRESUMEN
ETHNOPHARMACOLOGY RELEVANCE: Zanthoxylum bungeanum Maxim. (Z. bungeanum), a member of the Rutaceae family, has a rich history of traditional use in Asia for treating arthritis and toothache conditions. As characteristic chemical components, numerous kinds of alkaloids have been extracted from plants and their diverse biological activities have been reported. However, research on the isoquinoline alkaloid, a specific type of alkaloids, in Z. bungeanum was scarce. AIM OF THE STUDY: The study aimed to isolate a novel isoquinoline alkaloid from Z. bungeanum and explore its pharmacological activity in vitro and analgesic activity in vivo. MATERIALS AND METHODS: Isoquinoline alkaloid isolation and identification from Z. bungeanum were conducted using chromatographic and spectroscopic methods. The whole-cell patch-clamp technique was applied to assess its impact on neuronal excitability, and endogenous voltage-gated potassium (Kv) and sodium (Nav) currents in acutely isolated mouse small-diameter dorsal root ganglion (DRG) neurons. Its inhibitory impacts on channels were further validated with HEK293 cells stably expressing Nav1.7 and Nav1.8, and Chinese hamster ovary (CHO) cells transiently expressing Kv2.1. The formalin inflammatory pain model was utilized to evaluate the potential analgesic activity in vivo. RESULTS: A novel isoquinoline alkaloid named HJ-69 (N-13-(3-methoxyprop-1-yl)rutaecarpine) was isolated and identified from Z. bungeanum for the first time. HJ-69 significantly suppressed the firing frequency and amplitudes of action potentials in DRG neurons. Consistently, it state-dependently inhibited endogenous Nav currents of DRG neurons, with half maximal inhibitory concentration (IC50) values of 13.06 ± 2.06 µM and 30.19 ± 2.07 µM for the inactivated and resting states, respectively. HJ-69 significantly suppressed potassium currents in DRG neurons, which notably inhibited the delayed rectifier potassium (IK) currents (IC50 = 6.95 ± 1.29 µM) and slightly affected the transient outward potassium (IA) currents (IC50 = 523.50 ± 39.16 µM). Furtherly, HJ-69 exhibited similar potencies on heterologously expressed Nav1.7, Nav1.8, and Kv2.1 channels, which correspondingly represent the main components in neurons. Notably, intraperitoneal administration of 30 mg/kg and 100 mg/kg HJ-69 significantly alleviated pain behaviors in the mouse inflammatory pain model induced by formalin. CONCLUSION: The study concluded that HJ-69 is a novel and active isoquinoline alkaloid, and the inhibition of Nav and Kv channels contributes to its analgesic activity. HJ-69 may be a promising prototype for future analgesic drug discovery based on the isoquinoline alkaloid.
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Analgésicos , Ganglios Espinales , Dolor , Zanthoxylum , Animales , Zanthoxylum/química , Humanos , Células HEK293 , Analgésicos/farmacología , Analgésicos/química , Analgésicos/aislamiento & purificación , Analgésicos/uso terapéutico , Ganglios Espinales/efectos de los fármacos , Ganglios Espinales/metabolismo , Ratones , Masculino , Dolor/tratamiento farmacológico , Isoquinolinas/farmacología , Isoquinolinas/aislamiento & purificación , Isoquinolinas/química , Alcaloides/farmacología , Alcaloides/aislamiento & purificación , Alcaloides/química , Alcaloides/uso terapéutico , Bloqueadores de los Canales de Potasio/farmacología , Canal de Sodio Activado por Voltaje NAV1.7/metabolismo , Inflamación/tratamiento farmacológico , Bloqueadores del Canal de Sodio Activado por Voltaje/farmacología , Bloqueadores del Canal de Sodio Activado por Voltaje/aislamiento & purificación , Canales de Potasio con Entrada de Voltaje/metabolismo , Canales de Potasio con Entrada de Voltaje/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Antiinflamatorios/farmacología , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/química , Ratones Endogámicos C57BL , CricetulusRESUMEN
Prolyl hydroxylase domain 2 (PHD2) is an important enzyme in the human body that perceives changes in oxygen concentration and regulates response in hypoxic environments. Evaluation of PHD2 inhibitory activity of natural products is crucial for drug development of hypoxia related diseases. At present, the detection of low concentration of α-ketoglutaric acid (the substrate of PHD2 enzymatic reaction) requires derivatization reactions or sample pretreatment, which undoubtedly increases the workload of PHD2 inhibitory activity evaluation. In this paper, a direct detection approach of α-ketoglutaric acid was established by using the online stacking strategy of capillary electrophoresis to evaluate the PHD2 inhibitory activity of natural products. Under optimized conditions, detection of a single sample can be achieved within 2 min. By calculation, the intraday precision RSD of the apparent electrophoretic mobility and peak areas of α-ketoglutaric acid are 0.92 % and 0.79 %, respectively, and the interday RSD were 1.27 % and 0.96 % respectively. The recoveries of the present approach were 97.9-105.2 %, and the LOQ and LOD were 2.0 µM and 5.0 µM, respectively. Furthermore, this approach was applied for the evaluation of inhibitory activity of PHD2 for 13 natural products, and PHD2 inhibitory activity of salvianolic acid A was firstly reported. The present work not only realizes evaluation of PHD2 inhibitory activity through direct detection of α-ketoglutaric acid, but also provides technical support for the discovery of potential drug molecules in hypoxia related diseases.
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Productos Biológicos , Electroforesis Capilar , Prolina Dioxigenasas del Factor Inducible por Hipoxia , Ácidos Cetoglutáricos , Humanos , Productos Biológicos/farmacología , Electroforesis Capilar/métodos , Prolina Dioxigenasas del Factor Inducible por Hipoxia/antagonistas & inhibidores , Ácidos Cetoglutáricos/análisisRESUMEN
Microglia are resident macrophages within the central nervous system, serving as the first responders to neuroinflammation. Glucocorticoids (GCs) may cause damage to brain tissue, but the specific mechanism remains unclear. This study was divided into two parts: a glucocorticoid receptor (GR) mitochondrial translocation intervention experiment and a mitochondrial oxidative stress inhibition experiment. BV-2 microglia were stimulated with dexamethasone (DEX) and treated with either tubastatin-A or mitoquinone (MitoQ) for 24 h. Our results showed that DEX increased the translocation of GRs to mitochondria, and this effect was accompanied by decreases in the expression of mitochondrially encoded cytochrome c oxidase 1 (MT-CO1) and mitochondrially encoded cytochrome c oxidase 3 (MT-CO3) and increases in the expression of NOD-like receptor thermal protein domain-associated protein 3 (NLRP3), caspase-1, and Gasdermin D (GSDMD). The level of mitochondrial respiratory chain complex IV (MRCC IV) and adenosine triphosphate (ATP) was decreased. An elevation in the level of mitochondrial oxidative stress and the opening of the mitochondrial permeability transition pore (mPTP) was also observed. Mechanistically, tubastatin-A significantly suppressed the mitochondrial translocation of GRs, improved the expression of mitochondrial genes, promoted the restoration of mitochondrial function, and inhibited pyroptosis. MitoQ significantly prevented mitochondrial oxidative stress, improved mitochondrial function, and reduced apoptosis and pyroptosis. Both tubastatin-A and MitoQ suppressed DEX-induced pyroptosis. This study substantiates that the increase in the mitochondrial translocation of GRs mediated by GCs exacerbates oxidative stress and pyroptosis in microglia, which indicates that the regulation of mitochondrial pathways by GCs is pathogenic to microglia.
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Glucocorticoides , Piroptosis , Glucocorticoides/farmacología , Glucocorticoides/metabolismo , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Microglía/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Estrés Oxidativo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismoRESUMEN
BACKGROUND: Ocular trauma is a major cause of vision loss, especially in the young patients, and is the leading cause of unilateral blind in China. OBJECTIVE: The aims of this report are to analyze ciliary and choroidal lesion characteristics and outcomes of a group of patients with ruptured globe injuries and discuss finding a more effective treatment protocol. Here we report our experience treating ruptured globe injuries. METHODS: Seventy-five patients (75 eyes) with a diagnosis of ruptured globe injuries were selected from 264 patients with open globe injuries at the Shierming Eye Hospital of Shandong Province between January 2009 and December 2011. General information and clinical characteristics such as ciliary and choroidal lesion features were reviewed. RESULTS: Of the 75 patients, 85.3% were men, and the average age of the patients was 37.2 years (range, 6-63 years). The right eye was injured in 52.0%; enucleation was performed in 9 patients. There was no light perception, in the final corrected visual acuity in another 3 patients. The ratio of better visual acuity (better than 0.1) increased from 0 preoperatively to 16.0% postoperatively. Among the 75 patients with ruptured globe injuries, 13 had ciliary injury and 47 (62.7%) had choroidal injuries. Both ciliary and choroidal injuries were detected in 15 patients. Retinal tissue incarceration during sclera suturing was usually the vital point leading to unfavorable results. CONCLUSIONS: Ruptured globe injury usually results in severe visual acuity damage. Active treatment could help to restore visual acuity in patients to some degree. Some effective treatment protocols for ruptured globe injuries could be followed. Some unsuitable procedures in primary treatment should be avoided to achieve a better prognosis.