RESUMEN
Diabetic foot ulcer complicated with lower extremity vasculopathy is highly prevalent, slow healing and have a poor prognosis. The final progression leads to amputation, or may even be life-threatening, seriously affecting patients' quality of life. The treatment of lower extremity vasculopathy is the focus of clinical practice and is vital to improving the healing process of diabetic foot ulcers. Recently, a number of clinical trials on diabetic foot ulcers with lower extremity vasculopathy have been reported. A joint group of Chinese Medical Association (CMA) and Chinese Medical Doctor Association (CMDA) expert representatives reviewed and reached a consensus on the guidelines for the clinical diagnosis and treatment of this kind of disease. These guidelines are based on evidence from the literature and cover the pathogenesis of diabetic foot ulcers complicated with lower extremity vasculopathy and the application of new treatment approaches. These guidelines have been put forward to guide practitioners on the best approaches for screening, diagnosing and treating diabetic foot ulcers with lower extremity vasculopathy, with the aim of providing optimal, evidence-based management for medical personnel working with diabetic foot wound repair and treatment.
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Diabetes Mellitus , Pie Diabético , Úlcera del Pie , Glutamatos , Compuestos de Mostaza Nitrogenada , Humanos , Pie Diabético/complicaciones , Pie Diabético/diagnóstico , Pie Diabético/terapia , Consenso , Calidad de Vida , Extremidad InferiorRESUMEN
OBJECTIVES: We systematically reviewed published clinical trials to evaluate the effectiveness of virtual reality (VR) technology on functional improvement, pain relief, and reduction of mental distress among burn patients undergoing rehabilitation. DATA SOURCES: Systematic searches were conducted in 4 databases, including PubMed, the Cochrane Library, Embase, and Web of Science, from inception to August 2021. STUDY SELECTION: Randomized controlled trials (RCTs) evaluating any type of VR for the rehabilitation in burn patients with dysfunction were included. DATA EXTRACTION: Two reviewers evaluated the eligibility, and another 2 reviewers used the Cochrane risk of bias assessment tool to assess the risk of bias. The extracted data included the main results of rehabilitation evaluation (quality of life [QOL], work performance, range of motion [ROM] of joints, hand grip and pinch strength, pain, fun, anxiety), the application performance of VR (realness and presence), adverse effects (fatigue and nausea), and characteristics of the included studies. Heterogeneity was evaluated using the chi-square tests and I2 statistics. Random- or fixed-effects models were conducted to pool the effect sizes expressed as standardized mean differences (SMDs). DATA SYNTHESIS: Sixteen RCTs with 535 burn patients were included. VR-based interventions were superior to usual rehabilitation in QOL and work performance of burn patients and produced positive effect on the average gain of ROM (SMD=0.72) as well. VR was not associated with improved hand grip and pinch strength (SMD=0.50, 1.22, respectively) but was associated with reduced intensity, affective, and cognitive components of pain (SMD=-1.26, -0.71, -1.01, respectively) compared with control conditions. Ratings of fun in rehabilitation therapy were higher (SMD=2.38), and anxiety scores were lower (SMD=-0.73) than in control conditions. CONCLUSIONS: VR-based burn rehabilitation significantly improves the QOL and work performance of burn patients, increases the ROM gain in the joints, reduces the intensity and unpleasantness of pain and the time spent thinking about pain, increases the fun in the rehabilitation therapy, reduces the anxiety caused by the treatment, and has no obvious adverse effects. However, it did not significantly improve hand grip or pinch strength.
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Quemaduras , Realidad Virtual , Humanos , Quemaduras/rehabilitación , Dolor , Manejo del Dolor/métodos , Calidad de VidaRESUMEN
BACKGROUND: Diabetic nephropathy (DN) involves various structural and functional changes because of chronic glycemic assault and kidney failure. Proteinuria is an early clinical manifestation of DN, but the associated pathogenesis remains elusive. This study aimed to investigate the role of microtubule associated protein 4 (MAP4) phosphorylation (p-MAP4) in proteinuria in DN and its possible mechanisms. METHODS: In this study, the urine samples of diabetic patients and kidney tissues of streptozotocin (STZ)-induced diabetic mice were obtained to detect changes of p-MAP4. A murine model of hyperphosphorylated MAP4 was established to examine the effect of MAP4 phosphorylation in DN. Podocyte was applied to explore changes of kidney phenotypes and potential mechanisms with multiple methods. RESULTS: Our results demonstrated elevated content of p-MAP4 in diabetic patients' urine samples, and increased kidney p-MAP4 in streptozocin (STZ)-induced diabetic mice. Moreover, p-MAP4 triggered proteinuria with aging in mice, and induced epithelial-to-mesenchymal transition (EMT) and apoptosis in podocytes. Additionally, p-MAP4 mice were much more susceptible to STZ treatment and showed robust DN pathology as compared to wild-type mice. In vitro study revealed high glucose (HG) triggered elevation of p-MAP4, rearrangement of microtubules and F-actin filaments with enhanced cell permeability, accompanied with dedifferentiation and apoptosis of podocytes. These effects were significantly reinforced by MAP4 hyperphosphorylation, and were rectified by MAP4 dephosphorylation. Notably, pretreatment of p38/MAPK inhibitor SB203580 reinstated all HG-induced pathological alterations. CONCLUSIONS: The findings indicated a novel role for p-MAP4 in causing proteinuria in DN. Our results indicated the therapeutic potential of MAP4 in protecting against proteinuria and related diseases. Video Abstract.
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Diabetes Mellitus Experimental , Nefropatías Diabéticas , Podocitos , Animales , Diabetes Mellitus Experimental/metabolismo , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Transición Epitelial-Mesenquimal , Ratones , Proteínas Asociadas a Microtúbulos , Fosforilación , Podocitos/patología , Proteinuria/complicaciones , Estreptozocina/farmacologíaRESUMEN
Autophagy is a highly conserved self-protection mechanism that plays a crucial role in cardiovascular diseases. Cardiomyocyte hypoxic injury promotes oxidative stress and pathological alterations in the heart, although the interplay between these effects remains elusive. The transient receptor potential vanilloid 1 (TRPV1) ion channel is a nonselective cation channel that is activated in response to a variety of exogenous and endogenous physical and chemical stimuli. Here, we investigated the effects and mechanisms of action of TRPV1 on autophagy in hypoxic cardiomyocytes. In this study, primary cardiomyocytes isolated from C57 mice were subjected to hypoxic stress, and their expression of TRPV1 and adenosine 5'-monophosphate-activated protein kinase (AMPK) was regulated. The autophagy flux was assessed by Western blotting and immunofluorescence staining, and the cell viability was determined through Cell counting kit-8 assay and Lactate dehydrogenase assays. In addition, the calcium influx after the upregulation of TRPV1 expression in cardiomyocytes was examined. The results showed that the number of autophagosomes in cardiomyocytes was higher under hypoxic stress and that the blockade of autophagy flux aggravated hypoxic damage to cardiomyocytes. Moreover, the expression of TRPV1 was induced under hypoxic stress, and its upregulation by capsaicin improved the autophagy flux and protected cardiomyocytes from hypoxic damage, whereas the silencing of TRPV1 significantly attenuated autophagy. Our observations also revealed that AMPK signaling was activated and involved in TRPV1-induced autophagy in cardiomyocytes under hypoxic stress. Overall, this study demonstrates that TRPV1 activation mitigates hypoxic injury in cardiomyocytes by improving autophagy flux through the AMPK signaling pathway and highlights TRPV1 as a novel therapeutic target for the treatment of hypoxic cardiac disease.
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Autofagia/genética , Lesiones Cardíacas/genética , Proteínas Quinasas/genética , Canales Catiónicos TRPV/genética , Quinasas de la Proteína-Quinasa Activada por el AMP , Animales , Calcio/metabolismo , Capsaicina/farmacología , Supervivencia Celular/genética , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/genética , Lesiones Cardíacas/patología , Humanos , Hipoxia/genética , Hipoxia/metabolismo , Hipoxia/patología , Ratones , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Transducción de Señal/genéticaRESUMEN
Endogenous wound electric fields (EFs), an important and fundamental occurrence of wound healing, profoundly influence the directed migration of keratinocytes. Although numerous studies have unveiled the signals responsible for EF-biased direction, the mechanisms by which EFs promote keratinocyte motility remains to be elucidated. In our study, EFs enhanced the directed migratory speed of keratinocytes by inducing autophagic activity, thereby facilitating skin barrier restoration. Initially, we found that electrical signals directed keratinocytes to the cathode with enhanced motility parameters [ i.e., trajectory distance, trajectory speed, displacement distance, and displacement speed ( Td/ t)] and more efficient migration (directionality and Td/ t along the x axis, among others). Meanwhile, EFs induced a time-dependent increase in autophagic activity in keratinocytes, with constant autophagic flux, accompanied by increased transcription of numerous autophagy-related genes. Deficiency in Atg5, a key protein necessary for autophagosome formation, led to significant reduction of autophagy, which was accompanied by a substantial reduction in EF-stimulated directed motility. These results demonstrated a causal relationship between autophagy and EF-directed migratory speed. In addition, both cell migration under normal conditions and EF-biased directionality were autophagy independent. Thus, our findings define autophagy as an important functional regulator of electrically enhanced directed motility, adding to a growing understanding of EFs.-Yan, T., Jiang, X., Lin, G., Tang, D., Zhang, J., Guo, X., Zhang, D., Zhang, Q., Jia, J., Huang, Y. Autophagy is required for the directed motility of keratinocytes driven by electric fields.
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Autofagia , Movimiento Celular , Campos Electromagnéticos , Queratinocitos/metabolismo , Animales , Proteína 5 Relacionada con la Autofagia/deficiencia , Proteína 5 Relacionada con la Autofagia/genética , Línea Celular , Células Cultivadas , Humanos , Queratinocitos/fisiología , Queratinocitos/efectos de la radiación , RatonesRESUMEN
BACKGROUND The treatment of elderly patients with severe burns is difficult and the mortality rate is high. The aim of this study was to investigate the epidemiological features of elderly patients with severe burns. MATERIAL AND METHODS Data from 109 elderly patients with severe burns between January 2009 and December 2018 were retrospectively analyzed. Demographic data, clinical characteristics, treatments, and outcomes were statistically analyzed. RESULTS Among the 109 elderly patients with severe burns, the male-to-female ratio was 1.73: 1.0. The median age of the elderly patients was 67 years, and the median total body surface area (TBSA) burned was 42%. Notably, 67.9% of burns occurred at home and most frequently occurred in summer (38.5%) and winter (28.4%); flame and flash burns predominated (83.4%). The incidence of inhalation injury was 35.8%, and pre-existing comorbidities were observed in approximately 51.4% of the patients. The median length of stay in the hospital per TBSA burned was 0.4 days. The mortality rate in the elderly patients was 24.8%, and the mortality rates in the ≥70% TBSA group, inhalation injury group, and patients with 3 or more pre-existing comorbidities were significantly higher than in the other groups. The risk of death increased with an increase in the number of pre-existing comorbidities (odds ratio: 2.222; 95% confidence interval: 1.174-4.205). CONCLUSIONS At a major burn center in Southwest China, the incidence and mortality of elderly patients with severe burns displayed no downward trend. There are etiological characteristics of these age groups that should be considered for prevention. Meanwhile, multidisciplinary treatment in a hospital and an increase in the social support for the elderly population might improve outcomes.
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Quemaduras/epidemiología , Quemaduras/mortalidad , Anciano , Anciano de 80 o más Años , Pueblo Asiatico , Unidades de Quemados/tendencias , China/epidemiología , Comorbilidad , Femenino , Hospitalización/tendencias , Humanos , Incidencia , Tiempo de Internación , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Extracellular pH strongly affects cellular metabolism and function. An acidic environment induced under pathological conditions leads to cardiomyocyte injury and dysfunction, but the underlying mechanisms are still poorly understood. Autophagy has been reported as a cytoprotective mechanism that maintains cellular metabolism and viability by removing misfolded proteins and damaged organelles. In our research, we found that acidic environments inhibit autophagosome formation in cardiomyocytes. Up-regulation of autophagic activity, however, ameliorates the cell injury induced by acidic treatments.We also found that acidic treatments reduce the level of α-tubulin acetylation, as detected by Western blot and immunofluorescence staining, and that the number of autophagosomes increase after up-regulating α-tubulin acetylation by Taxol, suggesting that α-tubulin acetylation may play an important role in acidic pH-induced changes in autophagy. Furthermore, an HDAC6 activity assay showed an increase in HDAC6 activity after acidic treatment and that inhibiting HDAC6 activity by tubastatin A or specific siRNA up-regulates α-tubulin acetylation and autophagosome formation. These data confirm that autophagy plays a protective role against acidic pH-induced cell injury and indicate that HDAC6-mediated α-tubulin acetylation is an important mechanism of acidic pH-dependent autophagy in cardiomyocytes.
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Autofagosomas/metabolismo , Miocitos Cardíacos/metabolismo , Tubulina (Proteína)/metabolismo , Acetilación/efectos de los fármacos , Animales , Autofagosomas/efectos de los fármacos , Autofagia/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Histona Desacetilasa 6/metabolismo , Inhibidores de Histona Desacetilasas/farmacología , Concentración de Iones de Hidrógeno , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Ratas Sprague-DawleyRESUMEN
Although parallel alignment of fibroblasts to the tension lines of scar has been evidenced in vivo, how scar contracture generates directional contraction remains largely unclear due to the lack of effective in vitro model. Fibroblast populated collagen lattice (FPCL), a widely used in vitro model, fails to mimic scar contracture since it produces concentric contraction with the random orientation of fibroblast. We hypothesized that a novel FPCL model with fibroblast alignment might produce directional contraction and then simulate scar contracture better. Here, we showed that although direct current electric fields (DCEFs) enabled fibroblasts aligned perpendicularly to the field vector, it also promoted electrotactic migration of fibroblast in FPCL. By contrast, biphasic pulse direct current electric fields (BPDCEFs), featured by reversal of the EF direction periodically, abolished the electrotactic migration, but induced fibroblast alignment in a pulse frequency dependent manner. Specifically, BPDCEF at a pulse frequency of 0.0002â¯Hz induced fibroblast alignment comparable to that induced by DCEF under the same field strength (300â¯mV/mm), leading to an enhanced contraction of FPCL along the direction of cell alignment. FPCL pretreated by BPDCEF showed an elliptical contraction whereas it was concentric in control FPCL. Further study revealed that F-actin redistributions acted as a key mechanism for the induction of fibroblasts alignment by BPDCEF. Cytochalasin D, an inhibitor of actin dynamics, abolished F-actins redistribution, and significantly suppressed the fibroblasts alignment and the directional contraction of FPCL. Importantly, BPDCEF significantly increased RhoA activity in fibroblasts, while this response was attenuated by C3 transferase pre-treatment, a potent inhibitor of RhoA, caused F-actin depolymerization and actin filament bundle randomly distributed. Taken together, our study suggests a crucial role for fibroblast orientation in scar contracture, and provides a novel FPCL model that may be feasible and effective for investigating scar contracture in vitro.
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Electricidad , Fibroblastos/citología , Modelos Biológicos , Andamios del Tejido , ADP Ribosa Transferasas/farmacología , Actinas/antagonistas & inhibidores , Actinas/genética , Actinas/metabolismo , Animales , Animales Recién Nacidos , Fenómenos Biomecánicos , Toxinas Botulínicas/farmacología , Movimiento Celular , Cicatriz/genética , Cicatriz/metabolismo , Cicatriz/patología , Colágeno/química , Citocalasina D/farmacología , Femenino , Fibroblastos/metabolismo , Expresión Génica , Masculino , Ratones , Ratones Endogámicos BALB C , Cultivo Primario de Células , Ratas , Piel/citología , Piel/metabolismo , Tensión Superficial , Proteínas de Unión al GTP rho/antagonistas & inhibidores , Proteínas de Unión al GTP rho/genética , Proteínas de Unión al GTP rho/metabolismo , Proteína de Unión al GTP rhoARESUMEN
BACKGROUND/AIMS: Regional hypoxia promptly develops after trauma because of microvascular injury and increased oxygen consumption. This acute hypoxia plays a positive role in early skin wound healing. One of the mechanisms underlying the beneficial effects of acute hypoxia on wound healing may be increased hypoxia-inducible factor-1 (HIF-1α) expression. HIF-1α may affect the wound-healing process through many aspects, including angiogenesis, metabolism, and extra-cellular matrix synthesis and remodelling. Epidermal stem cells (EpSCs) are important participants in wound repair; however, whether these cells are regulated by hypoxia is unclear. This study aimed to elucidate the regulatory mechanism by which hypoxia acts on EpSCs. METHODS: CCK8 assays, western blots and live cell station observation were employed to compare the viability, proliferation and motility of EpSCs cultured under normoxic conditions (21% O2) with those cultured under hypoxic conditions (2% O2). Moreover, we used FG-4592 (a prolyl hydroxylase inhibitor that stabilizes HIF-1α in normoxia), KC7F2 (a selective inhibitor of HIF-1α transcription) and siRNA against HIF-1α to regulate HIF-1α expression. RESULTS: Acute hypoxia caused EpSCs to switch from a quiescent state to an activated state with higher viability and motility, as well as an earlier proliferation peak. We demonstrated that the HIF-1 signalling pathway mediated hypoxia-induced activation of EpSCs. Finally, the in vivo experiments showed that exogenous FG-4592 effectively accelerates wound healing, shortens healing times and even induces epidermal hyperplasia. CONCLUSION: This study demonstrated that both hypoxia and exogenous FG-4592 improve EpSC proliferation and motility by stabilizing HIF-1α, and its results suggest that HIF-1α is an important target through which wound healing can be accelerated and that FG-4592 is a promising new drug for wound repair.
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Epidermis/efectos de los fármacos , Glicina/análogos & derivados , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Isoquinolinas/uso terapéutico , Estabilidad Proteica/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Animales , Hipoxia de la Célula/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Células Epidérmicas , Epidermis/metabolismo , Epidermis/patología , Glicina/farmacología , Glicina/uso terapéutico , Isoquinolinas/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Células Madre/citología , Células Madre/efectos de los fármacos , Células Madre/metabolismoRESUMEN
The migration of keratinocytes from wound margins plays a critical role in the re-epithelialization of skin wounds. Hypoxia occurs immediately after injury and acts as an early stimulus to initiate the healing processes. Although our previous studies have revealed that hypoxia promotes keratinocyte migration, the precise mechanisms involved remain unclear. Here, we found that BNIP3 expression was upregulated in hypoxic keratinocytes, and BNIP3 silencing suppressed hypoxia-induced cell migration. Additionally, hypoxia activated the focal adhesion kinase (FAK) pathway through upregulation of BNIP3, while FAK inhibition attenuated hypoxic keratinocyte migration. Here, we conclusively demonstrate a novel role for BNIP3 in hypoxia-induced keratinocyte migration. Furthermore, we provide a new perspective on the molecular mechanisms of wound healing and identify BNIP3 as a potential new molecular target for clinical treatments to enhance wound healing.
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Movimiento Celular , Quinasa 1 de Adhesión Focal/metabolismo , Hipoxia/metabolismo , Queratinocitos/fisiología , Proteínas de la Membrana/metabolismo , Proteínas Mitocondriales/metabolismo , Animales , Animales Recién Nacidos , Células Cultivadas , Ratones Endogámicos BALB C , Transducción de SeñalRESUMEN
F-actin rearrangement is an early event in burn-induced endothelial barrier dysfunction. HSP27, a target of p38 MAPK/MK2 pathway, plays an important role in actin dynamics through phosphorylation. The question of whether HSP27 participates in burn-related endothelial barrier dysfunction has not been identified yet. Here, we showed that burn serum induced a temporal appearance of central F-actin stress fibers followed by a formation of irregular dense peripheral F-actin in pulmonary endothelial monolayer, concomitant with a transient increase of HSP27 phosphorylation that conflicted with the persistent activation of p38 MAPK/MK2 unexpectedly. The appearance of F-actin stress fibers and transient increase of HSP27 phosphorylation occurred prior to the burn serum-induced endothelial hyperpermeability. Overexpressing phospho-mimicking HSP27 (HSP27(Asp)) reversed the burn serum-induced peripheral F-actin rearrangement with the augmentation of central F-actin stress fibers, and more importantly, attenuated the burn serum-induced endothelial hyperpermeability; such effects were not observed by HSP27(Ala), a non-phosphorylated mutant of HSP27. HSP27(Asp) overexpression also rendered the monolayer more resistant to barrier disruption caused by Cytochalasin D, a chemical reagent that depolymerizes F-actin specifically. Further study showed that phosphatases and sumoylation-inhibited MK2 activity contributed to the blunting of HSP27 phosphorylation during the burn serum-induced endothelial hyperpermeability. Our study identifies HSP27 phosphorylation as a protective response against burn serum-induced endothelial barrier dysfunction, and suggests that targeting HSP27 wound be a promising therapeutic strategy in ameliorating burn-induced lung edema and shock development.
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Actinas/metabolismo , Quemaduras/sangre , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Proteínas de Choque Térmico HSP27/metabolismo , Actinas/análisis , Adulto , Animales , Quemaduras/patología , Línea Celular , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Persona de Mediana Edad , Permeabilidad , Fosforilación , Proteínas Serina-Treonina Quinasas/metabolismo , Ratas , Fibras de Estrés/metabolismo , Fibras de Estrés/patología , Sumoilación , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
The integrity of the vascular barrier, which is essential to blood vessel homoeostasis, can be disrupted by a variety of soluble permeability factors during sepsis. Pigment epithelium-derived factor (PEDF), a potent endogenous anti-angiogenic molecule, is significantly increased in sepsis, but its role in endothelial dysfunction has not been defined. To assess the role of PEDF in the vasculature, we evaluated the effects of exogenous PEDF in vivo using a mouse model of cecal ligation and puncture (CLP)-induced sepsis and in vitro using human dermal microvascular endothelial cells (HDMECs). In addition, PEDF was inhibited using a PEDF-monoclonal antibody (PEDF-mAb) or recombinant lentivirus vectors targeting PEDF receptors, including adipose triglyceride lipase (ATGL) and laminin receptor (LR). Our results showed that exogenous PEDF induced vascular hyperpermeability, as measured by extravasation of Evan's Blue (EB), dextran and microspheres in the skin, blood, trachea and cremaster muscle, both in a normal state and under conditions of sepsis. In control and LR-shRNA-treated HDMECs, PEDF alone or in combination with inflammatory mediators resulted in activation of RhoA, which was accompanied by actin rearrangement and disassembly of intercellular junctions, impairing endothelial barrier function. But in ATGL-shRNA-treated HDMECs, PEDF failed to induce the aforementioned alterations, suggesting that PEDF-induced hyperpermeability was mediated through the ATGL receptor. These results reveal a novel role for PEDF as a potential vasoactive substance in septic vascular hyperpermeability. Furthermore, our results suggest that PEDF and ATGL may serve as therapeutic targets for managing vascular hyperpermeability in sepsis.
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Permeabilidad Capilar/fisiología , Proteínas del Ojo/sangre , Lipasa/metabolismo , Factores de Crecimiento Nervioso/sangre , Sepsis/fisiopatología , Serpinas/sangre , Adulto , Animales , Western Blotting , Permeabilidad Capilar/efectos de los fármacos , Células Cultivadas , Activación Enzimática/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Proteínas del Ojo/farmacología , Humanos , Lipasa/genética , Masculino , Ratones Endogámicos BALB C , Microscopía Confocal , Persona de Mediana Edad , Factores de Crecimiento Nervioso/farmacología , Interferencia de ARN , Receptores de Neuropéptido/genética , Receptores de Neuropéptido/metabolismo , Sepsis/sangre , Sepsis/enzimología , Serpinas/farmacología , Uniones Estrechas/efectos de los fármacos , Uniones Estrechas/metabolismo , Proteína de la Zonula Occludens-1/metabolismo , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
Microtubules of cardiac myocytes depolymerize after a hypoxic insult or treatment with colchicine. However, little attention has been paid to quantifying changes in microtubule distribution when using fluorescent images. We converted fluorescence images of labeled microtubules in H9C2 cardiac myocytes to grayscale images, then filtered the images to remove any noise, and used grayscale histograms to quantify features of the images. The results show that parameters such as the mean, variance, skewness, kurtosis, energy, and entropy can be used to quantitatively describe the distribution of microtubules in cells. Quantitative characteristics of microtubule distribution were similar after culturing cells under hypoxic conditions or after treatment with colchicine. These results parallel those described for neonatal rat cardiac myocytes following ischemia and hypoxia. In addition, we provide a method for internal segmentation of the cells, which revealed that microtubular depolymerization was more evident near the cell membrane following hypoxia or colchicine treatment.
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Procesamiento de Imagen Asistido por Computador/métodos , Microscopía Fluorescente , Microtúbulos/metabolismo , Miocitos Cardíacos/química , Animales , Línea Celular , Colchicina/metabolismo , Hipoxia , Miocitos Cardíacos/efectos de los fármacos , Ratas , Coloración y Etiquetado/métodos , Moduladores de Tubulina/metabolismoRESUMEN
Microtubule analysis is of significant value for a better understanding of normal and pathological cellular processes. Although immunofluorescence microscopic techniques have proven useful in the study of microtubules, comparative results commonly rely on a descriptive and subjective visual analysis. We developed an objective and quantitative method based on image processing and analysis of fluorescently labeled microtubular patterns in cultured cells. We used a multi-parameter approach by analyzing four quantifiable characteristics to compose our quantitative feature set. Then we interpreted specific changes in the parameters and revealed the contribution of each feature set using principal component analysis. In addition, we verified that different treatment groups could be clearly discriminated using principal components of the multi-parameter model. High predictive accuracy of four commonly used multi-classification methods confirmed our method. These results demonstrated the effectiveness and efficiency of our method in the analysis of microtubules in fluorescence images. Application of the analytical methods presented here provides information concerning the organization and modification of microtubules, and could aid in the further understanding of structural and functional aspects of microtubules under normal and pathological conditions.
RESUMEN
Wound healing is a complex but well-orchestrated tissue repair process composed of a series of molecular and cellular events conducted by various types of cells and extracellular matrix. Despite a variety of therapeutic strategies proposed to accelerate the healing of acute and/or chronic wounds over the past few decades, effective treatment of chronic nonhealing wounds still remains a challenge. Due to the recent advances in stem cell research, a dramatic enthusiasm has been drawn to the application of stem cells in regenerative medicine. Both embryonic and adult stem cells have prolonged self-renewal capacity and are able to differentiate into various tissue types. Nevertheless, use of embryonic stem cells is limited, owing to ethical concerns and legal restrictions. Adult stem cells, which could be isolated from bone marrow, umbilical cord blood, adipose tissue, skin and hair follicles,are being explored extensively to facilitate the healing of both acute and chronic wounds. The current article summarizes recent research on various types of stem cell-based strategies applied to improve wound healing. In addition, future directions of stem cell-based therapy in wound healing have also been discussed. Finally, despite its apparent advantages, limitations and challenges of stem cell therapy are discussed.
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Tratamiento Basado en Trasplante de Células y Tejidos , Medicina Regenerativa , Piel/patología , Trasplante de Células Madre , Cicatrización de Heridas , Heridas y Lesiones/patología , Tratamiento Basado en Trasplante de Células y Tejidos/tendencias , Matriz Extracelular/patología , Femenino , Humanos , Masculino , Medicina Regenerativa/tendencias , Piel/lesiones , Células Madre/patología , Heridas y Lesiones/terapiaRESUMEN
Although Mesenchymal Stem Cells (MSCs)-based therapy has been proposed as a promising strategy for the treatment of chronic lower-extremity ulcers, their optimal sources, amounts, and delivery methods are urgently needed to be determined. In this study, we compared the heterogeneity of the human MSCs derived from bone marrow (BMSCs), umbilical cord (UCMSCs), and adipose tissue (ADSCs) in accelerating wound healing and promoting angiogenesis and explored the underlying mechanism. Briefly, a diabetic rat model with a full-thickness cutaneous wound on the dorsal foot was developed. The wound was topically administered with three types of MSCs. Additionally, we carried out in vitro and in vivo analysis of the angiogenic properties of the MSCs. Moreover, the molecular mechanism of the heterogeneity of the MSCs derived from the three tissues was explored by transcriptome sequencing. When compared with the BMSCs- and UCMSCs-treated groups, the ADSCs-treated group exhibited markedly accelerated healing efficiency, characterized by increased wound closure rates, enhanced angiogenesis, and collagen deposition at the wound site. The three types of MSCs formed three-dimensional capillary-like structures and promoted angiogenesis in vitro and in vivo, with ADSCs exhibiting the highest capacity for tube formation and pro-angiogenesis. Furthermore, transcriptome sequencing revealed that ADSCs had higher expression levels of angiogenesis-associated genes. Our findings indicate that MSCs-based therapy accelerates the healing of ischemia- and diabetes-induced lower-extremity ulcers and that adipose tissue-derived MSCs might be ideal for therapeutic angiogenesis and treatment of chronic ischemic wounds.
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Diabetes Mellitus , Células Madre Mesenquimatosas , Humanos , Ratas , Animales , Angiogénesis , Úlcera/metabolismo , Neovascularización Fisiológica/genética , Células Madre Mesenquimatosas/metabolismo , Cicatrización de Heridas/genéticaRESUMEN
BACKGROUND: Previous reports on the treatment of sacral and ischial pressure injuries have not provided clear algorithms for surgical therapies. The objective of this study was to establish a reconstruction algorithm to guide the selection of an ideal free-style perforator flap that can be tailored to the defect in question. METHODS: We used 23 perforator flaps to reconstruct 14 sacral and 8 ischial defects in 22 patients over 5 years. A reconstruction algorithm system was developed based on the anatomical features of the perforator vessels (diameter, D; pulsatility [++â¼+++], P) and their position in the skin island (DPD) (ie, D+P+DPD). A perforator-based propeller flap was applied as the first-line choice; if this plan was not feasible, we applied an altered V-Y advancement model or another second-choice technique. RESULTS: All flaps survived, and only 1 patient experienced partial wound dehiscence, which healed by secondary intention. After an average follow-up period of 11.2 months, no patient experienced recurrence or infection. CONCLUSIONS: Free-style perforator flap selection is determined by pressure injury and the desired advantage of a specific approach. The use of free-style perforator-based propeller flaps allows a surgeon to transfer healthy tissue into the defect, shifts the suture line away from the bony prominence, and preserves additional future donor sites. In cases where unexpected variations are encountered, the V-Y advancement model or another technique can be used. The simplified surgical algorithm (D+P+DPD) can provide versatility and reliability, achieve a durable, natural esthetic outcome, and minimize injuries to future donor sites.
Asunto(s)
Algoritmos , Isquion , Colgajo Perforante , Úlcera por Presión , Humanos , Colgajo Perforante/irrigación sanguínea , Úlcera por Presión/cirugía , Masculino , Persona de Mediana Edad , Femenino , Adulto , Anciano , Procedimientos de Cirugía Plástica/métodos , Sacro/cirugía , Sacro/lesionesRESUMEN
Calcium homeostasis imbalance is one of the important pathological mechanisms in heart failure. Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2a), a calcium ATPase on the sarcoplasmic reticulum in cardiac myocytes, is a myocardial systolic-diastolic Ca2â +â homeostasis regulating enzyme that is not only involved in cardiac diastole but also indirectly affects cardiac myocyte contraction. SERCA2a expression was found to be decreased in myocardial tissue in heart failure, however, there are few reports on serum SERCA2a expression in patients with heart failure, and this study was designed to investigate whether serum SERCA2a levels are associated with the occurrence of adverse events after discharge in patients hospitalized with heart failure. Patients with heart failure hospitalized in the cardiovascular department of the Second Affiliated Hospital of Guangdong Medical University, China, from July 2018 to July 2019 were included in this study, and serum SERCA2a concentrations were measured; each enrolled patient was followed up by telephone after 6 months (6â ±â 1 months) for general post-discharge patient status. The correlation between serum SERCA2a levels and the occurrence of adverse events (death or readmission due to heart failure) after hospital discharge was assessed using multiple analysis and trend analysis. Seventy-one patients with heart failure were finally included in this study, of whom 38 (53.5%) were men and 33 (46.5%) were women (All were postmenopausal women). Multiple analysis revealed no correlation between serum SERCA2a levels and the occurrence of adverse events in the total study population and in male patients, but serum SERCA2a levels were associated with the occurrence of adverse outcome events after hospital discharge in female patients (ORâ =â 1.02, Pâ =â .047). Further analysis using a trend analysis yielded a 4.0% increase in the risk of adverse outcomes after hospital discharge for each unit increase in SERCA2a in female patients (ORâ =â 1.04; Pâ =â .02), while no significant difference was seen in men. This study suggests that serum SERCA2a levels at admission are associated with the occurrence of post-discharge adverse events in postmenopausal female patients hospitalized with heart failure.
Asunto(s)
Insuficiencia Cardíaca , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico , Humanos , Femenino , Masculino , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Terapia Genética , Alta del Paciente , Cuidados Posteriores , Insuficiencia Cardíaca/terapia , Miocitos Cardíacos , Calcio/metabolismoRESUMEN
This study used the trauma film paradigm to investigate different forms of posttrauma verbal processing relevant to the formation of intrusive memories. We designed 3 experiments to investigate verbal processing that could help to reduce the formation of posttraumatic intrusions. Experiments 1 and 2 looked at the effect of several forms of verbal processing, varied in emotional foci and vantage points, on the formation of posttraumatic intrusions. Experiment 3 utilized event-related potential (ERP) technology to control emotional focus and to further examine the effect of verbal processing from different vantage points. Data produced by Experiment 1 showed that the "what-focus" group had fewer intrusions than the "why-focus" group. Experiment 2 produced no significant difference between first- and third-person vantage points. Results from the last experiment showed the what-focus group was faster to judge the colors of the words in the emotional Stroop task, and the amplitude and latency of P2 for negative words were greater than neutral words in the what-focus group. Based on the results of the experiments, participants who were led to verbalize their traumatic experiences using the what-focus and the first-person vantage point ended up with fewer intrusions.