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An imbalanced system of angiogenesis-osteoblasts-osteoclasts is regarded as the main factor in bone remodeling dysfunction diseases or osseointegration loss. Osteoclast precursors are the key cells that accelerate bone-specific angiogenesis and maintain normal osteoblast and osteoclast function. Graphene oxide is an effective scaffold surface modification agent with broad application prospects in bone tissue engineering. However, the effect of graphene oxide on the interaction between osteoclasts and angiogenesis has not yet been elucidated. In this study, a rat calvarial defect model was established and treated with an electrochemically derived nanographene oxide (ENGO) hydrogel. Higher angiogenesis and platelet-derived growth factor (PDGF) B in preosteoclasts were observed in the ENGO group compared with that in the control group. Moreover, in vitro experiments demonstrate the efficacy of ENGO in substantially reducing the expression of the receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast-associated markers and inhibiting bone resorption activity. Additionally, ENGO enhances the secretion of the osteoclast-derived coupling factor PDGF-BB and promotes angiogenesis. Our investigation revealed the crucial role of isocitrate dehydrogenase 1 (IDH1) in the ENGO-mediated regulation of osteoclast differentiation and PDGF-BB secretion. The decreased expression of IDH1 reduces the level of histone lysine demethylase 7A (KDM7A) and subsequently increases the H3K9me2 level in the cathepsin K promoter region. In summary, we found that ENGO promotes angiogenesis by inhibiting the maturity of RANKL-induced osteoclasts and enhancing PDGF-BB secretion. These results indicate that ENGO holds promise for the application in fostering osteoclast-endothelial cell crosstalk, providing an effective strategy for treating bone resorption and osteoclast-related bone loss diseases.
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Diferenciación Celular , Grafito , Neovascularización Fisiológica , Osteoclastos , Animales , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Diferenciación Celular/efectos de los fármacos , Ratas , Grafito/química , Grafito/farmacología , Neovascularización Fisiológica/efectos de los fármacos , Ratas Sprague-Dawley , Ratones , Masculino , Becaplermina/farmacología , Células Cultivadas , Isocitrato Deshidrogenasa/metabolismo , AngiogénesisRESUMEN
The etiology of gastrointestinal (GI) diseases is intricate and multifactorial, encompassing complex interactions between genetic predisposition and gut microbiota. The cell fate change, immune function regulation, and microenvironment composition in diseased tissues are governed by microorganisms and mutated genes either independently or through synergistic interactions. A comprehensive understanding of GI disease etiology is imperative for developing precise prevention and treatment strategies. However, the existing models used for studying the microenvironment in GI diseases-whether cancer cell lines or mouse models-exhibit significant limitations, which leads to the prosperity of organoids models. This review first describes the development history of organoids models, followed by a detailed demonstration of organoids application from bench to clinic. As for bench utilization, we present a layer-by-layer elucidation of organoid simulation on host-microbial interactions, as well as the application in molecular mechanism analysis. As for clinical adhibition, we provide a generalized interpretation of organoid application in GI disease simulation from inflammatory disorders to malignancy diseases, as well as in GI disease treatment including drug screening, immunotherapy, and microbial-targeting and screening treatment. This review draws a comprehensive and systematical depiction of organoids models, providing a novel insight into the utilization of organoids models from bench to clinic.
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Obesity is a risk factor for colorectal cancer (CRC), and the involvement of gut microbiota in the pathogenesis of obesity and CRC is widely recognized. However, the landscape of fecal microbiome and metabolome distinguishing patients with obesity-related CRC from obesity remains unknown. Here, we utilize metagenomic sequencing and metabolomics from 522 patients with CRC and healthy controls to identify the characteristics of obese CRC. Our integrated analysis reveals that obesity-related CRC is characterized by elevated Peptostreptococcus stomatis, dysregulated fatty acids and phospholipids, and altered Kyoto Encyclopedia of Genes and Genomes pathways involving glycerophospholipid metabolism and lipopolysaccharide synthesis. Correlation analysis unveils microbial interactions in obesity, where the probiotic Faecalibacterium prausnitzii and the tumor-promoting species P. stomatis may engage in cross-feeding, thereby promoting tumorigenesis. In vitro experiments affirm enhanced growth under cross-feeding conditions. The mutualistic microbe-microbe interaction may contribute to the association between obesity and elevated CRC risk. Additionally, diagnostic models incorporating BMI-specific microbial biomarkers display promise for precise CRC screening.
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Neoplasias Colorrectales , Microbiota , Humanos , Metaboloma , Obesidad/metabolismo , Neoplasias Colorrectales/microbiología , Interacciones MicrobianasRESUMEN
Chronic hepatitis B virus (HBV) infection is a major global public health risk that threatens human life and health, although the number of vaccinated people has increased. The clinical outcome of HBV infection depends on the complex interplay between viral replication and the host immune response. Innate immunity plays an important role in the early stages of the disease but retains no long-term immune memory. However, HBV evades detection by the host innate immune system through stealth. Therefore, adaptive immunity involving T and B cells is crucial for controlling and clearing HBV infections that lead to liver inflammation and damage. The persistence of HBV leads to immune tolerance owing to immune cell dysfunction, T cell exhaustion, and an increase in suppressor cells and cytokines. Although significant progress has been made in HBV treatment in recent years, the balance between immune tolerance, immune activation, inflammation, and fibrosis in chronic hepatitis B remains unknown, making a functional cure difficult to achieve. Therefore, this review focuses on the important cells involved in the innate and adaptive immunity of chronic hepatitis B that target the host immune system and identifies treatment strategies.
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Virus de la Hepatitis B , Hepatitis B Crónica , Inmunidad Innata , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/inmunología , Hepatitis B Crónica/terapia , Células Dendríticas/inmunología , Células Asesinas Naturales/inmunología , Monocitos/inmunología , Macrófagos/inmunología , Linfocitos T Reguladores/inmunología , Linfocitos B Reguladores/inmunología , HumanosRESUMEN
OBJECTIVE: To explore the potential osteogenic induction mechanism of enamel matrix derivatives (EMDs) on bone marrow mesenchymal stem cell (BMSC) sheets with different titanium surface morphologies. METHODS: The BMSCs were inoculated on the surfaces of titanium alloys with different morphologies: anodic oxidation (AO), sand-blasted, large grit and acid-etched, and no treatment (control). The proliferation and osteogenic differentiation of BMSCs on the different surface morphologies were observed with the same concentration of EMDs. To further understand the osteogenic mechanism of EMDs on BMSC sheets with different morphologies, a real-time RT-PCR and a western blot were used to detect the overall levels of osteogenic genes and osteogenic proteins. Finally, to verify the osteogenic effect of BMSC sheets stimulated by EMDs in vivo, BMSC sheets with different morphologies were implanted into the subcutaneous tissue of the back of nude mice, and the bone formation was detected by HE staining. RESULTS: The EMDs and surface morphology in the AO group synergically increased the expression levels of osteogenic active factors (RUNX2, OSX and OCN) and enhanced the osteogenic differentiation effect of BMSCs. The in vivo experiments showed that the BMSC sheets in the AO group were rich in osteogenic active factors, and promoted the formation of ectopic bone tissue after implantation into the subcutaneous tissue of the back of nude mice. CONCLUSION: EMDs and AO morphology synergically enhance the secretion of bone osteogenic active factors of BMSCs and promote the formation of heterotopic bone.
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BACKGROUND: Endoplasmic reticulum stress (ER stress) is activated in chronic sinusitis with nasal polyps (CRSwNP) and leads to increased expression of C/EBP homologous protein (CHOP). However, the role of CHOP in the pathogenesis of CRSwNP remains unclear. METHODS: CHOP expression was detected by immunohistochemistry staining in nasal mucosa of control and CRSwNP patients. Co-localization of CHOP and cleaved caspase3, p-MLKL, and CD68 was detected by immunofluorescence staining in CRSwNP patients. TNFα, IFNγ, IL1ß, LPS, and tunicamycin were added to primary dispersed nasal polyp cells (DNPCs) to explore their roles in cell death. Western blot, CCK8 assay, and flow cytometry were employed to detect cell death. RESULTS: CHOP was specifically activated in CRSwNP compared to controls. It was mainly macrophages that highly expressed CHOP, some of which underwent apoptosis and the other underwent necroptosis. IL1ß induced increased CHOP and apoptosis, and a slight p-MLKL. In addition, ER stress could also promote p-MLKL expression. Whereas classical TNFα plus IFNγ and LPS did not induce increased necroptosis in DNPCs. CONCLUSION: IL1ß induced the apoptotic pathway and minor necroptosis. And ER stress also plays a role in the occurrence of necroptosis in CRSwNP.
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Necroptosis , Sinusitis , Factor de Transcripción CHOP , Humanos , Lipopolisacáridos , Macrófagos/metabolismo , Macrófagos/patología , Sinusitis/metabolismo , Sinusitis/patología , Factor de Necrosis Tumoral alfa , Factor de Transcripción CHOP/metabolismoRESUMEN
Japanese encephalitis (JE) is a naturally occurring localized disease caused by the Japanese encephalitis virus, which is spread by the Culex tritaeniorhynchus. China has a high rate of JE. Shanxi, located in North China, has a high prevalence of adult JE. Adult JE has more severe complications, mortality, and a higher disease burden, making it a public health issue. This retrospective study examined the dynamic epidemic changes, high-risk areas of JE, and clinical characteristics and prognostic factors of adult JE in Shanxi Province. The findings revealed that July to September was the primary epidemic season of JE and that JE cases were mainly in individuals over the age of 40. The incidence of JE from 2005 to 2022 demonstrated a positive spatial correlation with significant clustering characteristics, with high-incidence clusters in the south and southeast. Multivariate logistic regression analysis revealed that higher cerebrospinal fluid pressure, higher white blood cell counts, higher neutrophil percentage, deep coma, and lower albumin were independent factors for poor prognosis of adult JE. The developed risk prediction model holds great promise in early prognosis assessment of patients, providing a basis for clinical decision-making and early clinical intervention.
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Encefalitis Japonesa , Adulto , Humanos , Encefalitis Japonesa/diagnóstico , Encefalitis Japonesa/epidemiología , Pronóstico , Estudios Retrospectivos , China/epidemiologíaRESUMEN
Fall armyworm Spodoptera frugiperda, a native insect pest in tropical and subtropical America, has rapidly spread to most parts of China and become a major pest of corn and other crops since invading in early January 2019. As an emergency and important control measure, chemical control of S. frugiperda has the advantages of quick effect and low cost. However, long-term and large-scale use of pesticides might pollute the environment and increase pest resistance. By improving the control effect and reducing the dosage of chemical pesticides, graphene oxide (GO) is used synergistically with insecticides to increase control efficacy to achieve low-cost and sustainable management of insect pests as a new type of synergist. In this study, graphene oxide was compounded with insecticides to form nanocomposites. To clarify pest physiological responses, the laboratory toxicity of graphene oxide-insecticide nanocomposites was measured on the larvae of S. frugiperda. The results demonstrated that GO could enhance the activity of four selected pesticides: chlorantraniliprole (Chl), beta cypermethrin (Bet), methoxyhydrazide (Met) and spinetoram (Spi). Compared with pesticides alone, the toxicity of Chl-GO, Bet-GO, Met-GO and Spi-GO mixtures to the third instar larvae of S. frugiperda increased by 1.56, 1.54, 2.53 and 1.74 times, respectively. The easy preparation and higher bioactivity of GO-pesticide nanocomposites indicated their promising application potential in pest control.
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To study the effect of miR-153-3p on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in a high glucose environment and its potential mechanism. The results showed that high glucose inhibited the osteogenic differentiation of BMSCs, and the expression of miR-153-3p increased during osteogenic differentiation. Further experiments found that in BMSCs induced by high glucose, overexpression of miR-153-3p inhibited the osteogenic differentiation of BMSCs, and the expressions of osteogenesis-related genes bone sialoprotein, Collagen I and alkaline phosphatase were down-regulated, while silencing of miR-153-3p alleviated the inhibition effect. The dual-luciferase reporter gene assay confirmed that the 3'-untranslated region (3'-UTR) of runt related transcription factor 2 (RUNX2) had a targeted binding site with miR-153-3p and a negative regulatory effect. Molecular studies further confirmed that miR-153-3p inhibited the osteogenic differentiation of BMSCs by targeting the 3'-UTR of RUNX2. In conclusion, our study found that as one key regulator of high glucose affecting the osteogenic differentiation of BMSCs, miR-153-3p may play a negative regulatory role by inhibiting the expression of RUNX2.
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Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex (Tc). The chi60 and chi70 chitinase genes are located on the gene cluster encoding Tc toxins. To clarify the insecticidal activity of chitinases and their relationship with Tc toxins, the insecticidal activity of the chitinases was assessed on Helicoverpa armigera. Then, the chi60 and chi70 genes of X. nematophila HB310 were knocked out by the pJQ200SK suicide plasmid knockout system. The insecticidal activity of Tc toxin from the wild-type strain (WT) and mutant strains was carried out. The results demonstrate that Chi60 and Chi70 had an obvious growth inhibition effect against the second instar larvae of H. armigera with growth-inhibiting rates of 81.99% and 90.51%, respectively. Chi70 had a synergistic effect with the insecticidal toxicity of Tc toxins, but Chi60 had no synergistic effect with Tc toxins. After feeding Chi60 and Chi70, the peritrophic membrane of H. armigera became inelastic, was easily broken and leaked blue dextran. The Δchi60, Δchi70 and Δchi60-chi70 mutant strains were successfully screened. The toxicity of Tc toxins from the WT, Δchi60, Δchi70 and Δchi60-chi70 was 196.11 µg/mL, 757.25 µg/mL, 885.74 µg/mL and 20,049.83 µg/mL, respectively. The insecticidal activity of Tc toxins from Δchi60 and Δchi70 was 3.861 and 4.517 times lower than that of Tc toxins from the WT, respectively, while the insecticidal activity of Tc toxins from the Δchi60-chi70 mutant strain almost disappeared. These results indicate that the presence of chi60 and chi70 is indispensable for the toxicity of Tc toxins.
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Quitinasas , Insecticidas , Mariposas Nocturnas , Toxinas Biológicas , Xenorhabdus , Animales , Proteínas Bacterianas/farmacología , Proteínas Bacterianas/toxicidad , Quitinasas/genética , Quitinasas/metabolismo , Humanos , Insecticidas/metabolismo , Insecticidas/farmacología , Larva , Toxinas Biológicas/metabolismo , Xenorhabdus/genéticaRESUMEN
Digestive system diseases arise primarily through the interplay of genetic and environmental influences; there is an urgent need in elucidating the pathogenic mechanisms of these diseases and deploy personalized treatments. Traditional and long-established model systems rarely reproduce either tissue complexity or human physiology faithfully; these shortcomings underscore the need for better models. Organoids represent a promising research model, helping us gain a more profound understanding of the digestive organs; this model can also be used to provide patients with precise and individualized treatment and to build rapid in vitro test models for drug screening or gene/cell therapy, linking basic research with clinical treatment. Over the past few decades, the use of organoids has led to an advanced understanding of the composition of each digestive organ and has facilitated disease modeling, chemotherapy dose prediction, CRISPR-Cas9 genetic intervention, high-throughput drug screening, and identification of SARS-CoV-2 targets, pathogenic infection. However, the existing organoids of the digestive system mainly include the epithelial system. In order to reveal the pathogenic mechanism of digestive diseases, it is necessary to establish a completer and more physiological organoid model. Combining organoids and advanced techniques to test individualized treatments of different formulations is a promising approach that requires further exploration. This review highlights the advancements in the field of organoid technology from the perspectives of disease modeling and personalized therapy.
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COVID-19 , Enfermedades del Sistema Digestivo , Enfermedades del Sistema Digestivo/tratamiento farmacológico , Enfermedades del Sistema Digestivo/genética , Humanos , Organoides , Medicina de Precisión/métodos , SARS-CoV-2/genéticaRESUMEN
INTRODUCTION: The influence of enamel matrix derivative (EMD) on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was explored in high glucose (HG) microenvironment with interaction of Wnt/ß-catenin pathway. MATERIALS AND METHODS: Extraction of BMSCs from Sprague-Dawley rats, culture, and identification were manifested. The cells were treated with different concentration of EMD in HG to figure out the most available concentration for proliferation and osteogenic differentiation. Then, observation of cell growth curve and cell cycle changes, and detection of Osterix, runt-related transcription factor 2 (Runx2), COL-I, early osteogenic indexes, Calcium salt deposition, and ß-catenin protein in Wnt/ß-catenin pathway were assured. After adding Wnt/ß-catenin pathway inhibitor (XAV-939) in the cells with osteogenesis induction, detection of binding of ß-catenin to Osterix was clarified. RESULTS: Via identification BMSCs cultured in vitro was qualified. Different concentrations of EMD could accelerate cell proliferation in HG and osteogenesis induction, and 75 µg/mL EMD had the best effect. The HG augmented BMSCs proliferation and the propidium iodide index of flow cytometry cycle was elevated in HG, which were strengthened via the EMD. After BMSCs' osteogenesis induction, Osterix, Runx2, CoL-1, early osteogenic indexes, and calcium salt deposition were reduced, but elevated via EMD. ß-Catenin was the lowest in the HG, but elevated after EMD. After addition of XAV-939, reduction of ß-catenin and the downstream (Osterix and Runx2) were manifested. Detection of binding protein bands was in ß-catenin and Osterix of the HG after EMD treatment. CONCLUSION: EMD may facilitate the osteogenic differentiation of BMSCs via activating the Wnt/ß-catenin pathway in HG.
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Células Madre Mesenquimatosas , Osteogénesis , Vía de Señalización Wnt , Animales , Células de la Médula Ósea/metabolismo , Calcio/metabolismo , Diferenciación Celular , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Glucosa/farmacología , Ratas , Ratas Sprague-Dawley , beta Catenina/metabolismoRESUMEN
The main aim of this research work was to develop and evaluate a drug delivery system with compression coating technology to control drug release at a constant rate. The compression coated tablets (CCTs) consist of the hydrophilic matrix core and the hydrophobic waxy coating. The presence of hydrophobic waxy coating could provide sufficient time for hydration of the core to prevent initial burst release. The mechanism research revealed that erosion was the main way of drug release and the releasing area was constant during the entire release process because the core tablet was located in the cup-shaped coating after one side cover was dropped at the lag time. This made the release behavior exhibit zero-order kinetics (R2>0.99). The coating rupture strength and the core swelling force at the lag time influenced erosion rate thus affecting release rate. Different solubility of drugs (propranolol hydrochloride, melatonin, and nifedipine) was selected as model drugs and the properties of the prepared CCTs in terms of formulations and in vitro release were evaluated. The release rate was independent of solubility, medium pH, and osmotic pressure. This zero-order controlled system could be applied to both controlled drug delivery and chrono pharmaceutical drug delivery.
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Celulosa , Tecnología , Preparaciones de Acción Retardada , Solubilidad , ComprimidosRESUMEN
Insect glutathione-S-transferases (GSTs) play essential roles in metabolizing endogenous and exogenous compounds. GSTs that are uniquely expressed in antennae are assumed to function as scavengers of pheromones and host volatiles in the odorant detection system. Based on this assumption, antennae-specific GSTs have been identified and functionally characterized in increasing number of insect species. In the present study, 17 putative GSTs were identified from the antennal transcriptomic dataset of the Indian meal moth, Plodia interpunctella, a severe stored-grain pest worldwide. Among the GSTs, only PiGSTd1 is antennae-specific according to both Fragments Per Kilobase Million (FPKM) and quantitative real-time PCR (qRT-PCR) analysis. Sequence analysis revealed that PiGSTd1 has a similar identity as many delta GSTs from other moths. Enzyme kinetic assays using 1-chloro-2,4-dinitrobenzene (CDNB) as substrates showed that the recombinant PiGSTd1 gave a K m of 0.2292 ± 0.01805 mM and a V max of 14.02 ± 0.2545 µmol·mg-1·min-1 under the optimal catalytic conditions (35°C and pH = 7.5). Further analysis revealed that the recombinant PiGSTd1 could efficiently degrade the sex pheromone component Z9-12:Ac (75.63 ± 5.52%), as well as aldehyde volatiles, including hexanal (89.10 ± 2.21%), heptanal (63.19 ± 5.36%), (E)-2-octenal (73.58 ± 3.92%), (E)-2-nonenal (75.81 ± 1.90%), and (E)-2-decenal (61.13 ± 5.24%). Taken together, our findings suggest that PiGSTd1 may play essential roles in degrading and inactivating a variety of odorants, especially sex pheromones and host volatiles of P. interpunctella.
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This study was set out to determine the function of LAMC2 in laryngeal cancer (LC). Initially, we identified the expression of LAMC2 in LC cells and tissues using TCGA datasets, GEO datasets (GSE143224), qRT-PCR, and western blot. Besides, we analyzed the correlations between LAMC2 and clinicopathologic features in LC patients. The CCK-8 assays were performed to detect cell viability and the half-maximal inhibitory concentration of cetuximab (IC50) in LC cells. We explored the correlations between LAMC2 and EGFR and further explored the regulation mechanism of cetuximab in LC. This study identified a high expression of LAMC2 in LC cells and tissues. The expression levels of LAMC2 were associated with TNM classification, lymph node (LN) metastasis, differentiation, and overall survival (OS). LAMC2 significantly promoted cell proliferation and cell viability. Besides, cetuximab significantly inhibited LAMC2 expression levels. LAMC2 significantly reversed the effect of cetuximab suppressing cell proliferation in LC cells. In conclusion, LAMC2 may act as a novel anti-cancer target in LC.
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Cetuximab , Laminina/genética , Neoplasias Laríngeas , Línea Celular Tumoral , Proliferación Celular , Cetuximab/farmacología , Humanos , Neoplasias Laríngeas/tratamiento farmacológico , Neoplasias Laríngeas/genéticaRESUMEN
BACKGROUND: Systemic inflammation via host-tumor interactions is currently recognized as the seventh cancer hallmark. The purpose of this study was to detect whether pretreatment peripheral indexes were associated with aggressive behavior and prognosis of laryngeal carcinoma patients. METHODS: The pretreatment peripheral indexes such as albumin and systematic immune-inflammation index (SII) in 338 patients with laryngeal carcinoma were retrospectively recorded, the relationships between them and clinicopathological features and prognosis were analyzed. RESULTS: A high SII value was significantly positively associated with age (P = 0.01), N stage (P = 0.022) and tumor differentiation (P = 0.001). A low albumin value was significantly negatively associated with age (P = 0.01), tumor location (P = 0.001) and T stage (P = 0.015), N stage (P = 0.001) and tumor differentiation (P = 0.001). Univariate and multivariate survival analysis showed that a high SII (HR: 2.415, 95% CI 1.400-4.184; P = 0.002), a low blood albumin content (HR: 3.194, 95% CI 2.030-5.025; P = 0.001) independently predicted poor overall survival (OS). However, neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and platelet distribution width (RDW) were not independent prognostic factors. CONCLUSION: Pretreatment peripheral indexes SII and albumin could function as inexpensive indicators of aggressive behavior and be feasible and promising predictive biomarkers for prognosis in laryngeal carcinoma patients. Quantification of pretreatment SII and albumin may help physicians to design more effective management and follow-up strategies in laryngeal carcinoma patients.
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Carcinoma , Neutrófilos , Albúminas , Humanos , Inflamación , Pronóstico , Estudios RetrospectivosRESUMEN
Serotonin receptor 6 (5-HT6R), a typical G protein-coupled receptor (GPCR) mainly expressed in the neurogenic area with constitutive activity, is of particular interest as a promising target for emotional impairment. Here, we found that 5-HT6R was highly expressed in human NSCs and activation of the receptor promoted self-renewal of human NSCs, and thus induced the expansion and folding of human cerebral organoids; dysfunction of receptor or inhibition of its constitutive activity resulted in the premature differentiation of NSCs, which ultimately depleted the NSC pool. The following mechanistic study revealed that EPAC-CREB signaling was involved in 5-HT6R regulation. Furthermore, we showed that mice with genetic deletion of 5-HT6R or knockin A268R mutant presented depression-like behaviors and impaired hippocampal neurogenesis for progressive decrease of the NSC pool. Thus, this study indicates that the modulation of 5-HT6R and its constitutive activity may provide a therapeutic alternative to alleviate depression.
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Encéfalo/metabolismo , Depresión/patología , Organoides/metabolismo , Receptores de Serotonina/metabolismo , Animales , Encéfalo/citología , Proteína de Unión a CREB/metabolismo , Diferenciación Celular , Autorrenovación de las Células/efectos de los fármacos , Etilaminas/farmacología , Edición Génica , Humanos , Indoles/farmacología , Ratones , Ratones Transgénicos , Mutagénesis Sitio-Dirigida , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neurogénesis , Organoides/citología , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Receptores de Serotonina/química , Receptores de Serotonina/genética , Transducción de Señal , Proteínas de Unión al GTP rab1/metabolismoRESUMEN
PirAB toxin was initially found in the Photorhabdus luminescens TT01 strain and is a demonstrated binary toxin with high insecticidal activity. In this paper, we co-expressed the pirAB gene of Xenorhabdus nematophila HB310 in a prokaryotic expression system, and we found that the PirAB protein showed high hemocoel insecticidal activity against Galleria mellonella, Helicoverpa armigera and Spodoptera exigua. LD50 values were 1.562, 2.003 and 2.17 µg/larvae for G. mellonella, H. armigera, and S. exigua, respectively (p > 0.05). Additionally, PirAB-interaction proteins were identified from G. mellonella by 6 × His Protein Pulldown combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Of which, arylphorin of G. mellonella showed the highest matching rate. A protein domain conservative structure analysis indicated that arylphorin has three domains including Hemocyanin-N, Hemocyanin-M, and Hemocyanin-C. Among these protein domains, Hemocyanin-C has immune and recognition functions. Further, Hemocyanin-C domain of arylphorin was identified to interact with PirA but not PirB by Yeast two-hybrid system. These findings reveal, for the first time, new host protein interacting with PirAB. The identification of interaction protein may serve as the foundation for further study on the function and insecticidal mechanism of this binary toxin from Xenorhabdus.
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Toxinas Bacterianas/farmacología , Proteínas de Insectos/metabolismo , Insecticidas/farmacología , Mariposas Nocturnas/efectos de los fármacos , Xenorhabdus/metabolismo , Animales , Toxinas Bacterianas/genética , Sitios de Unión , Cromatografía Liquida , Clonación Molecular , Proteínas de Insectos/química , Mariposas Nocturnas/clasificación , Mariposas Nocturnas/metabolismo , Unión Proteica , Dominios Proteicos , Espectrometría de Masas en Tándem , Técnicas del Sistema de Dos Híbridos , Xenorhabdus/genéticaRESUMEN
The intricate balance of neural stem cell (NSC) amplification and neurogenesis is central to nervous system development. Dopamine D1 receptor (DRD1) is a typical G protein-coupled receptor (GPCR) mainly expressed in neurogenic area, with high constitutive activity. The receptor appears in the embryonic period before the formation of mature synaptic contacts, which indicates that dopamine receptor and its constitutive activity play crucial roles in the embryonic brain development. Here, we found that DRD1 was enriched in human NSCs. Inhibition of the receptor activity by its inverse agonists promoted human NSCs proliferation and impeded its differentiation. These results were also mimicked by genetic knockdown of DRD1, which also blocked the effects of inverse agonists, suggesting a receptor-dependent manner. More interestingly, knock-in A229T mutant with reduced DRD1 constitutive activity by CRISPR-Cas9 genome editing technology resulted into increased endogenous human NSCs proliferation. These results were well reproduced in human cerebral organoids, and inhibition of the DRD1 constitutive activity by its inverse agonists induced the expansion and folding of human cerebral organoids. The anatomic analysis uncovered that decreasing the constitutive activity of DRD1 by its inverse agonists promoted the NSCs proliferation and maintenance that led to hindered cortical neurogenesis. Further mechanistic studies revealed that the PKC-CBP pathway was involved in the regulation by DRD1. Thus, our findings indicate that the constitutive activity of DRD1 and possibly other GPCRs plays an important role in the development of human nervous system.
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Encéfalo/metabolismo , Organoides/metabolismo , Receptores de Dopamina D1/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Encéfalo/citología , Diferenciación Celular/fisiología , Humanos , Células Madre Pluripotentes Inducidas/citología , Células Madre Pluripotentes Inducidas/metabolismo , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neurogénesis , Organoides/citologíaRESUMEN
RATIONALE: Cellular angiofibroma is a rare benign mesenchymal tumor which mostly occurs in the superficial soft tissues of the genital region. Occurrence in the head and neck region is extremely rare. To our knowledge, this is the first case of cellular angiofibroma arising in the hypopharynx. PATIENT'S CONCERNS: A 54-year-old male complained of a mass moving through his throat for 1 month. A tumor was found in the right lateral wall of the pharyngeal at the level of the epiglottis by laryngoscope. Magnetic resonance imaging confirmed the presence of a mass in the right lateral pharyngeal area. A benign tumor was suspected based on the clinical symptoms and imaging findings. DIAGNOSIS: A supporting laryngoscope was performed under general anesthesia and the lesion was resected. Immunohistochemical analysis revealed cellular angiofibroma. INTERVENTIONS: The patient underwent surgical excision of the lesion. OUTCOMES: Thus far, no recurrence has been observed 6 months after excision. LESSONS: Cellular angiofibroma located in the lateral pharyngeal is rare; however, immunohistochemical staining is helpful for its diagnosis. Treatment is relatively simple and requires local excision and follow-up.