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Heterotopic ossification (HO) occurs as a common complication after injury, while its risk factor and mechanism remain unclear, which restricts the development of pharmacological treatment. Clinical research suggests that diabetes mellitus (DM) patients are prone to developing HO in the tendon, but solid evidence and mechanical research are still needed. Here, we combined the clinical samples and the DM mice model to identify that disordered glycolipid metabolism aggravates the senescence of tendon-derived stem cells (TSCs) and promotes osteogenic differentiation. Then, combining the RNA-seq results of the aging tendon, we detected the abnormally activated autocrine CXCL13-CXCR5 axis in TSCs cultured in a high fat, high glucose (HFHG) environment and also in the aged tendon. Genetic inhibition of CXCL13 successfully alleviated HO formation in DM mice, providing a potential therapeutic target for suppressing HO formation in DM patients after trauma or surgery.
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Quimiocina CXCL13 , Glicolipídeos , Ossificação Heterotópica , Osteogênese , Receptores CXCR5 , Animais , Ossificação Heterotópica/metabolismo , Ossificação Heterotópica/patologia , Ossificação Heterotópica/genética , Camundongos , Humanos , Quimiocina CXCL13/metabolismo , Quimiocina CXCL13/genética , Glicolipídeos/metabolismo , Receptores CXCR5/metabolismo , Receptores CXCR5/genética , Células-Tronco/metabolismo , Tendões/metabolismo , Tendões/patologia , Masculino , Camundongos Endogâmicos C57BL , Diferenciação Celular , Senescência Celular , Transdução de Sinais , Células Cultivadas , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologiaRESUMO
Confocal fluorescence imaging of fine structures of the cell membrane is important for understanding their biofunctions but is often neglected due to the lack of an effective method. Herein, we develop new amphiphilic rhodamine fluorescent probe RMGs in combination with basal imaging for this purpose. The probes show high signal-to-noise ratio and brightness and low internalization rate, making them suitable for imaging the fine substructures of the cell membrane. Using the representative probe RMG3, we not only observed the cell pseudopodia and intercellular nanotubes but also monitored the formation of migrasomes in real time. More importantly, in-depth imaging studies on more cell lines revealed for the first time that hepatocellular carcinoma cells secreted much more adherent extracellular vesicles than other cell lines, which might serve as a potential indicator of liver cells. We believe that RMGs may be useful for investigating the fine structures of the cell membrane.
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Membrana Celular , Corantes Fluorescentes , Rodaminas , Corantes Fluorescentes/química , Rodaminas/química , Humanos , Membrana Celular/química , Imagem Óptica , Microscopia Confocal/métodos , Tensoativos/químicaRESUMO
Klebsiella aerogenes (K. aerogenes, KA) is a gram-negative opportunistic pathogen from the Klebsiella species and the Enterobacteriaceae family. However, the impact of K. aerogenes on colorectal cancer (CRC) remains uncertain. A colitis-associated tumorigenesis animal model was established by administering azoxymethane (AOM) and dextran sulfate sodium (DSS) to C57BL/6J mice. The concentration of K. aerogenes gavage in mice was 109 cfu. The study measured the following parameters: tumor formation (number and size), intestinal permeability (MUC2, ZO-1, and Occludin), colonic inflammation (TNF-α, IL-1ß, IL-6, and IL-10), proliferation and the fluctuation of the intestinal flora. Under the AOM/DSS-treated setting, K. aerogenes colonization worsened colitis by exacerbating intestinal inflammatory reaction and destroying the mucosal barrier. The intervention markedly augmented the quantity and dimensions of neoplasm in the AOM/DSS mice, stimulated cellular growth, and impeded cellular programmed cell death. In addition, K. aerogenes exacerbated the imbalance of the intestinal microbiota by elevating the abundance of Pseudomonas, Erysipelatoclostridium, Turicibacter, Rikenella, and Muribaculum and leading to a reduction in the abundance of Odoribacter, Alloprevotella, Roseburia, and Lachnospiraceae_NK4A136_group. The presence of K. aerogenes in AOM/DSS-treated mice promoted tumorigenesis, worsened intestinal inflammation, disrupted the intestinal barrier, and caused disturbance to the gut microbiota.
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Colite , Enterobacter aerogenes , Animais , Camundongos , Azoximetano/toxicidade , Azoximetano/metabolismo , Camundongos Endogâmicos C57BL , Colite/patologia , Colo/patologia , Inflamação/patologia , Carcinogênese/patologia , Transformação Celular Neoplásica/patologia , Bacteroidetes , Sulfato de Dextrana/toxicidade , Modelos Animais de DoençasRESUMO
Burn injury is a serious traumatic injury that leads to severe physical and psychosocial impairment. Wound healing after burn injury is a substantial challenge in medical community. This study investigated the biological effects of the demethylase fat mass and obesity-associated protein (FTO) on burn injury. FTO protein level in burn skin tissues of patients was measured with Western blot assay. Keratinocytes (HaCaT cells) were given heat stimulation to induce an in vitro burn injury model, and then transfected with overexpression plasmids of FTO (pcDNA-FTO) or small interfering RNA against FTO (si-FTO). Cell proliferation, migration, and angiogenesis in keratinocytes were evaluated with CCK-8, Transwell, and tube formation assays, respectively. Tissue factor pathway inhibitor-2 (TFPI-2) m6A methylation level was detected with MeRIPqPCR assay. Then rescue experiments were conducted to explore the effects of FTO/TFPI-2 axis on keratinocyte functions. Lentivirus carrying FTO overexpression plasmids was injected into a burn rat model to detect its effects on wound healing and depressive-like behaviors in burn rats. FTO was downregulated in burn skin and heat-stimulated keratinocytes. FTO prominently augmented proliferation, migration and angiogenesis in heat-stimulated keratinocytes, while FTO knockdown showed the opposite results. FTO inhibited TFPI-2 expression by FTO-mediated m6A methylation modification. TFPI-2 overexpression abrogated FTO mediated enhancement of proliferation, migration and angiogenesis in keratinocytes. Additionally, FTO overexpression accelerated wound healing and improved depressive-like behaviors in burn rat model. FTO prominently augmented proliferation, migration and angiogenesis in heat-stimulated keratinocytes though inhibiting TFPI-2, and then improved wound healing and depressive-like behaviors.
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Angiogênese , Queimaduras , Glicoproteínas , Animais , Humanos , Ratos , Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética , Queimaduras/genética , Proliferação de Células , Desmetilação , Depressão/genética , Queratinócitos , CicatrizaçãoRESUMO
Short-chain fatty acids (SCFAs) are the primary energy source of colonic epithelial cells, but oral SCFAs are digested, absorbed, or degraded before reaching the colon. The acylated starch with SCFAs can be fermented and release specific SCFAs under the action of colonic intestinal microbiota. This review first introduces the preparation method, reaction mechanism, and substitution factors. Second, the structure, physical and chemical properties, in vitro function, and mechanism of acylated starch were expounded. Finally, the application of acylated starch in foods is introduced, and its safety is evaluated, providing a basis for the further development of acylated starch-based foods. The acylated starch obtained by different acylation types and preparation methods is different in particle, molecular, and crystal structures, leading to changes in the function and physicochemical properties. Meanwhile, acylated starch has the functional potential of targeted delivery of SCFAs to the colon, which can increase SCFAs in feces and intestine, selectively regulate the intestinal microbiota, and produce a prebiotic effect conducive to host health. The safety of acetylated starch has been supported by relevant studies, which have been widely used in various food fields and have great potential in the food industry.
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PURPOSE: Nearly all patients with hip fractures undergo surgical treatment. The use of different anesthesia techniques during surgery may influence the clinical outcomes. The optimal anesthetic technique for patients undergoing hip fracture surgery is still controversial. We performed this updated systematic review and meta-analysis to compare clinical outcomes of patients undergoing hip fracture surgery with different anesthesia techniques. SOURCE: Articles published from 2000 to May 2023 were included from MEDLINE, Embase, Web of Science, and the Cochrane Library. We included randomized controlled trials and observational studies comparing general anesthesia (GA) with regional anesthesia (RA) for the outcomes of 30-day mortality, 90-day mortality, in-hospital mortality, perioperative complications, length of hospital stay, and length of surgery in patients undergoing hip fracture surgery. Subgroup analyses were performed for the outcomes based on study design (randomized controlled trials or observational studies). We used a random-effects model for all analyses. PRINCIPAL FINDINGS: In this meta-analysis, we included 12 randomized controlled trials. There was no difference in postoperative 30-day mortality between the two groups (odds ratio [OR], 0.88; 95% confidence interval [CI], 0.44 to 1.74; I2 = 0%). The incidence of intraoperative hypotension was lower in patients who received RA vs GA (OR, 0.52; 95% CI, 0.38 to 0.72; I2 = 0%). No significant differences were observed in 90-day mortality, in-hospital mortality, postoperative delirium, pneumonia, myocardial infarction, venous thromboembolism, length of surgery, and length of hospital stay. CONCLUSION: In this updated systematic review and meta-analysis, RA did not reduce postoperative 30-day mortality in hip fracture surgery patients compared to GA. Fewer patients receiving RA had intraoperative hypotension than those receiving GA did. Apart from intraoperative hypotension, the data showed no differences in complications between the two anesthetic techniques. STUDY REGISTRATION: PROSPERO (CRD42023411854); registered 7 April 2023.
RéSUMé: OBJECTIF: Presque toutes les personnes ayant subi une fracture de la hanche se font opérer. L'utilisation de différentes techniques d'anesthésie pendant la chirurgie peut influencer les issues cliniques. La technique d'anesthésie optimale pour la patientèle bénéficiant de chirurgie de fracture de la hanche est encore controversée. Nous avons réalisé cette mise à jour par revue systématique et méta-analyse pour comparer les issues cliniques des personnes bénéficiant d'une chirurgie de fracture de la hanche avec différentes techniques d'anesthésie. SOURCES: Les articles publiés de 2000 à mai 2023 ont été inclus à partir des bases de données MEDLINE, Embase, Web of Science et Cochrane Library. Nous avons inclus des études randomisées contrôlées et des études observationnelles comparant l'anesthésie générale (AG) à l'anesthésie régionale (AR) pour les issues de mortalité à 30 jours, de mortalité à 90 jours, de mortalité intrahospitalière, de complications périopératoires, de durée de séjour à l'hôpital et de durée de la chirurgie pour les personnes bénéficiant d'une chirurgie de fracture de la hanche. Des analyses de sous-groupes ont été réalisées pour les issues en fonction de la méthodologie utilisée (étude randomisée contrôlée ou étude observationnelle). Un modèle à effets aléatoires a été utilisé pour toutes les analyses. CONSTATATIONS PRINCIPALES: Dans cette méta-analyse, nous avons inclus 12 études randomisées contrôlées. Il n'y avait pas de différence dans la mortalité postopératoire à 30 jours entre les deux groupes (rapport de cotes [RC], 0,88; intervalle de confiance à 95 % [IC], 0,44 à 1,74; I2 = 0 %). L'incidence d'hypotension peropératoire était plus faible chez les patient·es ayant reçu une AR vs une AG (RC, 0,52; IC 95 %, 0,38 à 0,72; I2 = 0 %). Aucune différence significative n'a été observée dans les issues de mortalité à 90 jours, de mortalité intrahospitalière, de delirium postopératoire, de pneumonie, d'infarctus du myocarde, de thromboembolie veineuse, de durée de la chirurgie, et de durée du séjour à l'hôpital. CONCLUSION: Dans cette revue systématique avec méta-analyse, l'anesthésie régionale n'a pas réduit la mortalité postopératoire à 30 jours chez les personnes ayant bénéficié d'une chirurgie de fracture de la hanche par rapport à l'anesthésie générale. Une proportion moindre de patient·es ayant reçu une AR présentaient une hypotension peropératoire par rapport aux personnes ayant reçu une AG. En dehors de l'hypotension peropératoire, les données n'ont montré aucune différence dans les complications entre les deux techniques anesthésiques. ENREGISTREMENT DE L'éTUDE: PROSPERO (CRD42023411854); enregistrée le 7 avril 2023.
Assuntos
Anestesia por Condução , Anestesia Geral , Fraturas do Quadril , Mortalidade Hospitalar , Tempo de Internação , Complicações Pós-Operatórias , Ensaios Clínicos Controlados Aleatórios como Assunto , Humanos , Fraturas do Quadril/cirurgia , Anestesia Geral/métodos , Anestesia por Condução/métodos , Tempo de Internação/estatística & dados numéricos , Complicações Pós-Operatórias/epidemiologiaRESUMO
Intervertebral disc degeneration (IDD) is the cause of low back pain (LBP), and recent research has suggested that inflammatory cytokines play a significant role in this process. Maslinic acid (MA), a natural compound found in olive plants ( Olea europaea), has anti-inflammatory properties, but its potential for treating IDD is unclear. The current study aims to investigate the effects of MA on TNFα-induced IDD in vitro and in other in vivo models. Our findings suggest that MA ameliorates the imbalance of the extracellular matrix (ECM) and mitigates senescence by upregulating aggrecan and collagen II levels as well as downregulating MMP and ADAMTS levels in nucleus pulposus cells (NPCs). It can also impede the progression of IDD in rats. We further find that MA significantly affects the PI3K/AKT and NF-κB pathways in TNFα-induced NPCs determined by RNA-seq and experimental verification, while the AKT agonist Sc-79 eliminates these signaling cascades. Furthermore, molecular docking simulation shows that MA directly binds to PI3K. Dysfunction of the PI3K/AKT pathway and ECM metabolism has also been confirmed in clinical specimens of degenerated nucleus pulposus. This study demonstrates that MA may hold promise as a therapeutic agent for alleviating ECM metabolism disorders and senescence to treat IDD.
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Degeneração do Disco Intervertebral , NF-kappa B , Núcleo Pulposo , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Ratos Sprague-Dawley , Transdução de Sinais , Triterpenos , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/tratamento farmacológico , Degeneração do Disco Intervertebral/patologia , Animais , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , NF-kappa B/metabolismo , Núcleo Pulposo/metabolismo , Núcleo Pulposo/efeitos dos fármacos , Núcleo Pulposo/patologia , Masculino , Triterpenos/farmacologia , Ratos , Humanos , Simulação de Acoplamento Molecular , Fator de Necrose Tumoral alfa/metabolismo , Matriz Extracelular/metabolismo , Matriz Extracelular/efeitos dos fármacos , Feminino , Células Cultivadas , Ácido Oleanólico/análogos & derivadosRESUMO
Absorption spectra are fundamental bases for the qualitative and quantitative analysis of the target chemical, and the development of an analytical model can be improved by studying its characteristics and rules. In the present study, the electronic excitation characteristics of phosphate anions (H2PO4-, HPO42-, and PO43-) were analyzed based on the charge-transfer spectrum. In addition, the absorption spectra of phosphate anions at the energy level of PBE0/6-311+G (d,p) were recorded based on the time-dependent density functional theory (TD-DFT) method. Different (HPO42-)n·(H2O)10-n molecular clusters were theoretically constructed, and the combined TD-DFT method and independent gradient model revealed that red shift of the maximum absorption wavelength (λmax) with the increase of phosphate anion concentration (0-10 mM) may be caused by the decrease of hydrogen bond interaction. In addition, the prominent dispersion in the short-wave region mainly resulted in the red shift of λmax with the increase in optical path length (1-100 mm). Moreover, with the increase in spectral bandwidth (0.4-3.0 nm), λmax slightly blue-shifted because of the increase in energy through the slit, and repeatability of the corresponding absorbance measurement at λmax gradually improved. As the spectral bandwidth increased, light monochromaticity became poor, resulting in the decrease of the linearly fitted correlation coefficient of the concentration-absorbance curve. Finally, the multivariate analysis of variance results showed that the optical path length was the most significant factor that influenced the absorption spectral characteristics of phosphate anions. This study provides a basis for the qualitative and quantitative analysis of phosphate anions by using absorption spectra and also renders a theoretical reference for absorption spectroscopy of other chemicals.
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BACKGROUND: Mesenchymal stem cells (MSCs) possess the potential to differentiate into chondrocytes, which makes them an ideal source for healing cartilage defects. Here, we seek to identify the essential genes participating in MSCs chondrogenesis. METHODS: Human MSCs were induced for chondrogenesis for 7, 14, and 21 days using a high-density micromass culture system, and RNA was extracted for RNA-seq. RESULTS: A total of 6247 differentially expressed genes (DEGs) were identified on day 7, and 85 DEGs were identified on day 14. However, no significant DEGs was identified on day 21. The top 30 DEGs at day 7, including COL9A3, COL10A1, and CILP2, are closely related to extracellular matrix organization. While the top 30 DEGs at day 14 revealed that inflammation-related genes were enriched, including CXCL8, TLR2, and CCL20. We also conducted protein-protein interaction (PPI) networks analysis using the search tool for the retrieval of interacting genes (STRING) database and identified key hub genes, including CXCL8, TLR2, CCL20, and MMP3. The transcriptional factors were also analyzed, identifying the top 5 TFs: LEF1, FOXO1, RORA, BHLHE41, and SOX5. We demonstrated one particular TF, RORA, in promoting early MSCs chondrogenesis. CONCLUSIONS: Taken together, our results suggested that these DEGs may have a complex effect on MSCs chondrogenesis both synergistically and solitarily.
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Condrogênese , Células-Tronco Mesenquimais , Humanos , Condrogênese/genética , Receptor 2 Toll-Like , Diferenciação Celular/genética , Condrócitos , Células CultivadasRESUMO
BACKGROUND: With the widespread use of the posterior surgery, more and more surgeons chose posterior surgery to treat thoracic and lumbar tuberculosis. But others still believed that the anterior surgery is more conducive to eradicating the lesions, and easier to place larger bone pieces for bone graft fusion. We compared the clinical and radiological outcomes of anterior and posterior surgical approaches and presented our views. METHODS: This study included 52 thoracic and lumbar tuberculosis patients at Sun Yat-sen Memorial Hospital from January 2010 to June 2018. All cases underwent radical debridement, nerve decompression, intervertebral bone graft fusion and internal fixation. Cases were divided into anterior group (24 cases) and posterior group (28 cases). Statistical analysis was used to compare the clinical effectiveness, radiological outcomes, complications and other related information. RESULTS: Patients in the anterior group and the posterior group were followed up for an average of 27.4 and 22.3 months, respectively. There were no statistically significant differences between groups in the preoperative, postoperative and last follow-up VAS score, ASIA grade and Cobb angle of local kyphosis. Moreover, there were no statistically significant differences in the improvement of neurological function, loss of kyphotic correction, total incidence of complications, operative time, intraoperative blood loss and hospital stay between the two groups (P > 0.05). But there was greater correction of kyphosis, earlier bone fusion, lower incidence of poor wound healing, less interference with the normal spine and less internal fixation consumables and medical cost in the anterior group (P < 0.05). CONCLUSIONS: Both anterior and posterior approaches are feasible for thoracic and lumbar tuberculosis. While for thoracic and lumbar tuberculosis patients with a single lesion limited in the anterior and middle columns of the spine without severe kyphosis, the anterior approach surgery may have greater advantages in kyphosis correction, bone fusion, wound healing, protection of the normal spine, and medical consumables and cost.
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Cifose , Fusão Vertebral , Tuberculose da Coluna Vertebral , Estudos de Casos e Controles , Desbridamento , Humanos , Cifose/cirurgia , Vértebras Lombares/cirurgia , Estudos Retrospectivos , Vértebras Torácicas/cirurgia , Resultado do Tratamento , Tuberculose da Coluna Vertebral/cirurgiaRESUMO
The receptor tyrosine kinase MET plays a vital role in skeletal muscle development and in postnatal muscle regeneration. However, the effect of MET on myogenesis of myoblasts has not yet been fully understood. This study aimed to investigate the effects of MET on myogenesis in vivo and in vitro. Decreased myonuclei and down-regulated expression of myogenesis-related markers were observed in Met p.Y1232C mutant heterozygous mice. To explore the effects of MET on myoblast proliferation and differentiation, Met was overexpressed or interfered in C2C12 myoblast cells through the lentiviral transfection. The Met overexpression cells exhibited promotion in myoblast proliferation, while the Met deficiency cells showed impediment in proliferation. Moreover, myoblast differentiation was enhanced by the stable Met overexpression, but was impaired by Met deficiency. Furthermore, this study demonstrated that SU11274, an inhibitor of MET kinase activity, suppressed myoblast differentiation, suggesting that MET regulated the expression of myogenic regulatory factors (MRFs) and of desmin through the classical tyrosine kinase pathway. On the basis of the above findings, our work confirmed that MET promoted the proliferation and differentiation of myoblasts, deepening our understanding of the molecular mechanisms underlying muscle development.
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Diferenciação Celular , Proliferação de Células , Desenvolvimento Muscular , Mioblastos/citologia , Proteínas Proto-Oncogênicas c-met/metabolismo , Animais , Células Cultivadas , Camundongos , Mioblastos/metabolismoRESUMO
Intervertebral disc degenerative disease (IDD) is the most common degenerative spine disease, which leads to chronic low back pain and symptoms in the lower extremities. In this study, we found that RORα, a member of the retinoid-related orphan receptor family, is significantly elevated in nucleus pulposus tissue in IDD patients. The elevation of RORα is associated with increased apoptosis of nucleus pulposus (NP) cells. Therefore, we applicated a well-established inverse agonist of RORα, SR3335, to investigate its role in regulating NP cell metabolism and apoptosis. To further investigate the mechanism that SR3335 regulates the pathogenesis of IDD in vitro, tumor necrosis factor alpha (TNF-α) stimulation was used in human NP cells to mimic the hostile environment that leads to degeneration. We found that SR3335 treatment reversed the trend of increased apoptosis in NP cells induced by TNF-α treatment. Next, TNF-α treatment upregulated the expression of type II collagen and aggrecan and downregulated MMP13 (matrix-degrading enzyme matrix metalloproteinase 13) and ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4). However, these effects were reversed after SR3335 treatment. Furthermore, we find that SR3335 mediated the effect in NP cells by regulating the YAP signaling pathway, especially by affecting the phosphorylation state of YAP. In conclusion, the reduction of matrix degradation enzymes and apoptosis upon SR3335 treatment suggests that SR3335 is a promising drug in reversing the deleterious microenvironment in IDD patients.
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Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Núcleo Pulposo/metabolismo , Proteínas de Sinalização YAP/biossíntese , Proteína ADAMTS4/metabolismo , Idoso , Animais , Apoptose , Linhagem Celular Tumoral , Sobrevivência Celular , Colágeno Tipo II/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Pessoa de Meia-Idade , Simulação de Acoplamento Molecular , Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares/agonistas , Fosforilação , Ratos , Ratos Sprague-Dawley , Sulfonamidas/farmacologia , Tiofenos/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Regulação para CimaRESUMO
Protein glycosylation plays pivotal role in a variety of biological processes and has association with many diseases. The highly efficient glycopeptide enrichment is essential for the mass spectrometry-based glycoproteome research to reduce interference from non-glycopeptides. In this study, novel glutathione-functionalized two-dimensional cobalt sulfide nanosheets (Co-S@Au-GSH) were synthesized for rapid and highly effective enrichment of glycopeptides. By using this nanomaterial, 34 and 21 N-glycopeptides were effectively captured from human serum immunoglobulin G (IgG) and horseradish peroxidase (HRP) digests, respectively. In addition, the Co-S@Au-GSH showed remarkable performance in N-glycopeptide extraction with high selectivity (HRP: BSA = 1:500), low limit of detection (0.5 fmol/µL), high binding capacity (150 mg/g), good reusability, and great robustness. Moreover, it was successfully applied in complex serum samples, demonstrating its excellent enrichment performance. These results indicated that this nanomaterial has great potential in complicated practice samples in glycoproteome determination.
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Cobalto/química , Glutationa/química , Glicopeptídeos/isolamento & purificação , Nanocompostos/química , Fracionamento Químico/métodos , Glicopeptídeos/sangue , Peroxidase do Rábano Silvestre/sangue , Peroxidase do Rábano Silvestre/química , Peroxidase do Rábano Silvestre/isolamento & purificação , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/química , Imunoglobulina G/isolamento & purificação , Limite de Detecção , Fragmentos de Peptídeos/sangue , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/isolamento & purificação , Proteólise , Proteômica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
BACKGROUND: Pre-existing degeneration of adjacent segment is an important risk factor for adjacent-segment degeneration (ASD), but only limited and controversial studies have addressed its management. METHODS: We retrospectively analyzed patients with symptomatic degeneration of the L5/S1 segment warranting surgical interference and severe asymptomatic degeneration of the L4/5 segment. Of these patients, those who underwent interbody fusion of the causative (L5/S1) segment and distraction of the intervertebral space and facet fusion of the adjacent L4/5 segment were included in Group A (n = 103), while those who underwent interbody fusion of both the L5/S1 and L4/5 segments were included in Group B (n = 81). Clinical and radiographic outcomes were evaluated. RESULTS: Mean follow-up time was 58.5 months (range, 48-75 months). We found no significant difference in clinical outcomes or incidence of ASD in the L3/4 segment between Groups A and B. Compared with Group B, Group A experienced less bleeding (315 ± 84 ml vs. 532 ± 105 ml), shorter operation time (107 ± 34 min vs. 158 ± 55 min) and lower costs (US $13,830 ± $2640 vs. US $16,020 ± $3380; P < 0.05). In Group A, the disc height ratio (DHR) of the L4/5 segment was significantly increased from a preoperative value of 0.40 ± 0.13 to a last-follow-up value of 0.53 ± 0.18 (P < 0.05), while the degree of canal stenosis (DCS) was decreased from a preoperative value of 34.3 ± 11.2% to a last-follow-up value of 15.9 ± 9.3 % (P < 0.05). CONCLUSIONS: This modified method could be effective in treating severe asymptomatic pre-existing degeneration of adjacent segment in the lumbar spine.
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Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/cirurgia , Vértebras Lombares/diagnóstico por imagem , Fusão Vertebral , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Vértebras Lombares/cirurgia , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The excessive expression of cholinesterases (ChEs) directly disturbs the metabolism of acetylcholine (ACh), causing disordering neurotransmission in the brain or even Alzheimer's disease and cancer. However, the variation of ChEs including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in brain glioma has not yet been investigated. Therefore, the development of a suitable method for in situ imaging ChEs in brain tissues to understand the physiological functions of ChEs in depth is very important. Herein, a new near-infrared emission fluorescent probe (IPAN) for visualization of ChE activity was developed. IPAN exhibits ultrafast response to ChEs, low detection limit for AChE (0.127 U/mL) and BChE (0.0117 U/mL), and a large Stokes shift with near-infrared emission. Based on these excellent attributes, the IPAN was effectively utilized for imaging the fluctuations of ChE activity in the apoptosis cells and zebrafish. Notably, by utilizing the unique probe IPAN, we observed a significant enhancement of ChE activity in the tumor cells and brain glioma, for the first time. We believe that this interesting finding could provide a powerful guidance for tumor resection in the future.
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Acetilcolinesterase/metabolismo , Neoplasias Encefálicas/enzimologia , Corantes Fluorescentes/química , Glioma/enzimologia , Acetilcolinesterase/análise , Neoplasias Encefálicas/diagnóstico por imagem , Linhagem Celular , Corantes Fluorescentes/síntese química , Glioma/diagnóstico por imagem , Humanos , Estrutura Molecular , Imagem ÓpticaRESUMO
Propionyl-CoA carboxylase (PCC) is a promising enzyme in the fields of biological CO2 utilization, synthesis of natrual products, and so on. The activity and substrate specificity of PCC are dependent on its key subunit carboxyltransferase (CT). To obtain PCC with high enzyme activity, seven pccB genes encoding CT subunit from diverse microorganisms were expressed in recombinant E. coli, and PccB from Bacillus subtilis showed the highest activity in vitro. To further optimize this protein using directed evolution, a genetic screening system based on oxaloacetate availability was designed to enrich the active variants of PccBBs. Four amino acid substitutions (D46G, L97Q, N220I and I391T) proved of great assistance in PccBBs activity improvement, and a double mutant of PccBBs (N220I/I391T) showed a 94-fold increase of overall catalytic efficiency indicated by kcat/Km. Moreover, this PccBBs double mutant was applied in construction of new succinate biosynthetic pathway. This new pathway produces succinate from acetyl-CoA with fixation of two CO2 molecules, which was confirmed by isotope labeling experiment with NaH13CO3. Compared with previous succinate production based on carboxylation of phosphoenolpyruvate or pyruvate, this new pathway showed some advantages including higher CO2 fixation potentiality and availability under aerobic conditions. In summary, this study developed a PCC with high enzyme activity which can be widely used in biotechnology field, and also demonstrated the feasibility of new succinate biosynthetic pathway with two CO2 fixation reactions.
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Dióxido de Carbono , Ácido Succínico , Vias Biossintéticas , Escherichia coli/genética , Escherichia coli/metabolismo , Metilmalonil-CoA Descarboxilase/genética , Metilmalonil-CoA Descarboxilase/metabolismo , SuccinatosRESUMO
The excessive production of sulfur dioxide (SO2) and formaldehyde (FA) by organisms is closely associated with several diseases, including neurological disorders, respiratory disease, and genotoxicity. RNA plays a central role in the expression of genes. However, it is difficult for fluorescent probes to explore the intersecting correlation of SO2 and FA near the nucleolus regions due to the protective obstacle of the cell nucleus membrane. Herein, a unique fluorescent probe (DP) for reversible detection of the interaction of SO2 and FA in the nucleolus regions was constructed. In light of the outstanding attributes of DP, including a large fluorescence ratio variation, ultrafast detection of SO2, and excellent reversibility, it was successfully employed to track the variation of endogenous SO2 and FA in the cytoplasm and nucleolus regions of live cells. In addition, the DP probe could be applied to the imaging of endogenous SO2 and then recovered by FA in live mice.
Assuntos
Nucléolo Celular/metabolismo , Citoplasma/metabolismo , Fluorescência , Corantes Fluorescentes/química , Formaldeído/análise , Imagem Óptica/métodos , Dióxido de Enxofre/análise , Animais , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The purpose of this study was to investigate the effects of microRNA-196b (miRNA-196b) on proliferation, migration, invasiveness and apoptosis of hepatocellular carcinoma cell line (HepG2), and the mechanism involved. MiRNA-196b inhibitor or negative control were transfected into HepG2 cells, while empty liposome vector was used as normal control. The results of transfection were assessed using real-time quantitative polymerase chain reaction (qRT-PCR). Cell proliferation, migration, invasiveness and apoptosis were determined using cell counting kit 8 (CCK-8), scratch test, Transwell invasion assay, and flow cytometric analysis, respectively. The expressions of PIK3, Akt and p-Akt proteins were determined using Western blotting. The HepG2 cells were also treated with PI3K/Akt signaling pathway inhibitor LY294002, and its effect on cell proliferation, migration, invasion, and apoptosis, and expressions of PIK3, Akt, and p-Akt proteins were determined. The results of RT-PCR showed that the relative expression of miRNA-196b in the inhibitor group (0.42 ± 0.13) was significantly lower than that in the blank control group (0.96 ± 0.10) and the negative control group (1.01 ± 0.32) (p < 0.05). The miRNA-196b inhibitor significantly and time-dependently reduced the invasiveness, proliferation migration abilities of HepG2 cells, while promoting their apoptosis (p < 0.05). The expressions of PIK3 and p-Akt proteins were significantly down-regulated in the inhibitor group, when compared with normal and negative control groups (p < 0.05). However, there were no significant differences in the expression of Akt protein among the groups (p > 0.05). After treatment of HepG2 cells with PI3K/Akt signaling pathway inhibitor LY294002, the proliferative, migratory and invasive abilities of cells in the treatment group were significantly enhanced, while cell apoptosis was significantly reduced (p < 0.05). Similarly, the protein expressions of PIK3 and p-Akt in the non-treatment group was significantly upregulated, relative to the treatment group (p < 0.05), but there was no significant difference in the expression of Akt protein between the two groups (p > 0.05). Downregulation of miRNA-196b expression inhibits the proliferation, migration and invasiveness of HepG2 cells, while promoting their apoptosis via a mechanism involving the PI3K/Akt signaling pathway.
Assuntos
Apoptose/genética , Movimento Celular/genética , Regulação para Baixo/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Cromonas/farmacologia , Regulação para Baixo/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , MicroRNAs/metabolismo , Morfolinas/farmacologia , Invasividade Neoplásica , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
Food powders are ubiquitous in people's daily life and food industry production with numerous advantages. With the increasing use of food powder in the world, it is increasingly important to understand the processing methods of food powder and the changes in physicochemical properties after processing in order to control the processing conditions and the quality of final products. Among the many processing methods for food powders, superfine grinding technique is an emerging and useful tool for superfine powder manufacturing with some unique and promising properties, which has made the technique successfully applied in food processing over the recent decades. This review highlights the information of the main superfine grinding methods in the field of food processing and the changes in performance after superfine grinding treatment. Also, potential challenges, promising opportunities, and perspectives of this technology are covered in the review. This review will provide theoretical foundation and guideline for superfine food powder processing, which can provide new approaches and ideas for the high-value utilization of food raw materials as well as the food design.
Assuntos
Manipulação de Alimentos/métodos , Alimentos , Pós/química , Tamanho da PartículaRESUMO
Pancreatic cancer has a poor prognosis. Many epidemiological evidence show that diabetes is closely related to the occurrence of pancreatic cancer. The concentration of insulin in pancreas local tissues is higher than that in systemic circulation. In this study, we aimed to investigate the effect of insulin on pancreatic duct epithelial cells and identify the potential mechanisms. We found that insulin promoted the proliferation of pancreatic duct epithelial cells in the dependent of increased PLK1. Furthermore, PI3K/AKT and NF-κB pathway were involved in this process. By using PI3K/AKT inhibitor LY294002 and NF-κB shRNA, the increased PLK1 was reversed and cells proliferation was inhibited. Additionally, immunofluorescence analysis revealed the co-localization between PLK1 and ß-catenin. We showed that insulin can promote the increased expression of ß-catenin dependent on PLK1. This study showed that insulin may promotes cell proliferative vitality of pancreatic ductal epithelial cells by inducing PLK1 through PI3K/AKT and NF-κB pathway; The upregulation of PLK1 may reduce the degradation of ß-catenin. This may be one of the mechanisms by which T2DM promotes pancreatic cancer.