RESUMEN

Rheumatoid arthritis (RA) is a common autoimmune and inflammatory disease characterized by inflammation and hyperplasia of the synovial tissues. RA pathogenesis involves multiple cell types, genes, transcription factors (TFs) and networks. Yet, little is known about the TFs, and key drivers and networks regulating cell function and disease at the synovial tissue level, which is the site of disease. In the present study, we used available RNA-seq databases generated from synovial tissues and developed a novel approach to elucidate cell type-specific regulatory networks on synovial tissue genes in RA. We leverage established computational methodologies to infer sample-specific gene regulatory networks and applied statistical methods to compare network properties across phenotypic groups (RA versus osteoarthritis). We developed computational approaches to rank TFs based on their contribution to the observed phenotypic differences between RA and controls across different cell types. We identified 18 (fibroblast-like synoviocyte), 16 (T cells), 19 (B cells) and 11 (monocyte) key regulators in RA synovial tissues. Interestingly, fibroblast-like synoviocyte (FLS) and B cells were driven by multiple independent co-regulatory TF clusters that included MITF, HLX, BACH1 (FLS) and KLF13, FOSB, FOSL1 (B cells). However, monocytes were collectively governed by a single cluster of TF drivers, responsible for the main phenotypic differences between RA and controls, which included RFX5, IRF9, CREB5. Among several cell subset and pathway changes, we also detected reduced presence of Natural killer T (NKT) cells and eosinophils in RA synovial tissues. Overall, our novel approach identified new and previously unsuspected Key driver genes (KDG), TF and networks and should help better understanding individual cell regulation and co-regulatory networks in RA pathogenesis, as well as potentially generate new targets for treatment.

Asunto(s)

Artritis Reumatoide , Redes Reguladoras de Genes , Membrana Sinovial , Humanos , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Membrana Sinovial/metabolismo , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Perfilación de la Expresión Génica , Biología Computacional/métodos , Sinoviocitos/metabolismo , Osteoartritis/genética , Osteoartritis/metabolismo , Regulación de la Expresión Génica , Linfocitos B/inmunología , Linfocitos B/metabolismo , Transcriptoma

RESUMEN

Ferroptosis is a form of iron-dependent regulated cell death caused by the accumulation of lipid peroxides. In this review, we summarize research on the impact of ferroptosis on disease models and isolated cells in various types of arthritis. While most studies have focused on rheumatoid arthritis (RA) and osteoarthritis (OA), there is limited research on spondylarthritis and crystal arthropathies. The effects of inducing or inhibiting ferroptosis on the disease strongly depend on the studied cell type. In the search for new therapeutic targets, inhibiting ferroptosis in chondrocytes might have promising effects for any type of arthritis. On the other hand, ferroptosis induction may also lead to a desired decrease of synovial fibroblasts in RA. Thus, ferroptosis research must consider the cell-type-specific effects on arthritis. Further investigation is needed to clarify these complexities.

Asunto(s)

Ferroptosis , Osteoartritis , Humanos , Animales , Osteoartritis/metabolismo , Osteoartritis/patología , Condrocitos/metabolismo , Condrocitos/patología , Artritis Reumatoide/metabolismo , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Artritis/metabolismo , Artritis/patología , Fibroblastos/metabolismo , Fibroblastos/patología , Hierro/metabolismo

RESUMEN

Rheumatoid arthritis (RA) is a chronic condition characterized by inflammation and an abnormal immune response. N6-methyladenosine (m6A) methylation has altered nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing (NLRP) 3. This change is implicated in the regulation of cell pyroptosis and inflammation. WTAP has a crucial role in regulating NLRP3 m6A. In this work, we used a rat model of collagen-induced arthritis (CIA) to investigate the involvement of WTAP in the evolution of inflammation in RA. The purpose of silencing or overexpressing WTAP in RA-fibroblast-like synoviocytes (RA-FLSs) treated with TNF-α was to identify its impact on pyroptosis, NLRP3 inflammasome-related proteins, the secretion of pro-inflammatory cytokines and migration. Bioinformatics techniques were used to pinpoint the exact target controlled by WTAP. To assess WTAP and NLRP3's role in RA-FLSs, we used methylated RNA immunoprecipitation, LDH test, flow cytometry, RT-qPCR, Western blotting, and Transwell. Our results show that WTAP expression is upregulated in both RA rats and cell models. Cell pyroptosis, NLRP3-related pro-inflammatory cytokines, and migration were reduced in TNF-α-treated RA-FLSs when WTAP was knocked down, whereas overexpression of WTAP displayed the opposite effect in RA-FLSs. WTAP mediated m6A modification in the NLRP3 mRNA and enhanced its mRNA stability. These results suggested that WTAP promoted FLSs pyroptosis and related inflammatory response via NLRP3 and identified WTAP as a potential target for treating RA.

RESUMEN

BACKGROUND: Although Interferon-beta (IFNß) has long been approved as a disease-modifying therapy (DMT) for Multiple sclerosis (MS), flu-like syndrome (FLS) persists as a common adverse effect of interferon therapy. Given the importance of circadian rhythm in regulating physiological processes, we aimed to assess the relationship between patient's chronotype and time of interferon injection with FLS score in MS patients receiving IFNß. METHODS: A cross-sectional study was conducted on 118 MS patients who were referred to the clinic of neurology of Zanjan Vali-e-Asr Hospital for interferon injection. The included were invited to complete a morningness-eveningness questionnaire (MEQ) assessing patients' chronotype. The following data were extracted from patients' record: age, gender, duration of interferon treatment, type of interferon taken, time of interferon injection (morning/evening), FLS score, MS subtype, and usage of pain killers. All data found were imported and statistically analyzed in SPSS ver.26. RESULTS: According to the patients' record, 114 (96.6%) patients had experienced post-interferon injection FLS with different severities. Statistical analysis revealed no significant relationship between the patient's chronotype and FLS score. Nevertheless, the FLS score was significantly higher in those who had evening injections. CONCLUSIONS: Time of interferon injection was significantly associated with FLS score, with higher FLS score following evening injection. However, no significant relationship was found between the FLS score and the patient's chronotype. It is recommended that further studies assessing circadian rhythm using laboratory tests such as melatonin measurement need to be undertaken to investigate the association of circadian rhythm with post-interferon injection FLS.

Asunto(s)

Ritmo Circadiano , Interferón beta , Esclerosis Múltiple , Humanos , Femenino , Masculino , Adulto , Estudios Transversales , Interferón beta/efectos adversos , Interferón beta/administración & dosificación , Esclerosis Múltiple/tratamiento farmacológico , Ritmo Circadiano/fisiología , Ritmo Circadiano/efectos de los fármacos , Persona de Mediana Edad , Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/efectos adversos , Gripe Humana/complicaciones , Gripe Humana/tratamiento farmacológico , Adulto Joven , Índice de Severidad de la Enfermedad , Cronotipo

RESUMEN

FLAGELLIN SENSING 2 (FLS2) encodes a pattern recognition receptor that perceives bacterial flagellin. While putative FLS2 orthologs are broadly conserved in plants, their functional characterization remains limited. Here, we report the identification of orthologs in cucumber (Cucumis sativus) and melon (C. melo), named CsFLS2 and CmFLS2, respectively. Homology searching identified CsFLS2, and virus-induced gene silencing (VIGS) demonstrated that CsFLS2 is required for flg22-triggered ROS generation. Interestingly, genome re-sequencing of melon cv. Lennon and subsequent genomic PCR revealed that Lennon has two CmFLS2 haplotypes, haplotype I encoding full-length CmFLS2 and haplotype II encoding a truncated form. We show that VIGS-mediated knockdown of CmFLS2 haplotype I resulted in a significant reduction in both flg22-triggered ROS generation and immunity to a bacterial pathogen in melon cv. Lennon. Remarkably, genomic PCR of CmFLS2 revealed that 68% of tested commercial melon cultivars possess only CmFLS2 haplotype II: these cultivars thus lack functional CmFLS2. To explore evolutionary aspects of CmFLS2 haplotype II occurrence, we genotyped the CmFLS2 locus in 142 melon accessions by genomic PCR and analyzed 437 released sequences. The results suggest that CmFLS2 haplotype II is derived from C. melo subsp. melo. Furthermore, we suggest that the proportion of CmFLS2 haplotype II increased among the improved melo group compared with the primitive melo group. Collectively, these findings suggest that the deleted FLS2 locus generated in the primitive melo subspecies expanded after domestication, resulting in the spread of commercial melon cultivars defective in flagellin recognition, which is critical for bacterial immunity.

Asunto(s)

Flagelina , Proteínas de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Domesticación , Haplotipos , Cucurbitaceae/genética , Cucurbitaceae/microbiología , Cucurbitaceae/inmunología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Filogenia , Eliminación de Secuencia

RESUMEN

Infectious bronchitis virus (IBV) strains of the Delmarva (DMV)/1639 genotype have been causing false layer syndrome (FLS) in the Eastern Canadian layer operations since the end of 2015. FLS is characterized by the development of cystic oviducts in layer pullets infected at an early age. Currently, there are no homologous vaccines for the control of this IBV genotype. Our previous research showed that a heterologous vaccination regimen incorporating Massachusetts (Mass) and Connecticut (Conn) IBV types protects layers against DMV/1639 genotype IBV. The aim of this study was to investigate the role of maternal antibodies conferred by breeders received the same vaccination regimen in the protection against the development of DMV/1639-induced FLS in pullets. Maternal antibody-positive (MA+) and maternal antibody-negative (MA-) female progeny chicks were challenged at 1 day of age and kept under observation for 16 weeks. Oviductal cystic formations were observed in 3 of 14 birds (21.4 %) in the MA- pullets, while the lesions were notably absent in the MA+ pullets. Milder histopathological lesions were observed in the examined tissues of the MA+ pullets. However, the maternal derived immunity failed to demonstrate protection against the damage to the tracheal ciliary activity, viral shedding, and viral tissue distribution. Overall, this study underscores the limitations of maternal derived immunity in preventing certain aspects of viral pathogenesis, emphasizing the need for comprehensive strategies to address different aspects of IBV infection.

Asunto(s)

Anticuerpos Antivirales , Pollos , Infecciones por Coronavirus , Virus de la Bronquitis Infecciosa , Enfermedades de las Aves de Corral , Vacunas Virales , Animales , Virus de la Bronquitis Infecciosa/inmunología , Enfermedades de las Aves de Corral/prevención & control , Enfermedades de las Aves de Corral/inmunología , Enfermedades de las Aves de Corral/virología , Pollos/inmunología , Pollos/virología , Femenino , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Vacunas Virales/inmunología , Vacunas Virales/administración & dosificación , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/virología , Inmunidad Materno-Adquirida , Tráquea/inmunología , Tráquea/virología , Oviductos/inmunología , Oviductos/patología , Oviductos/virología

RESUMEN

Force is crucial for learning psychomotor skills in laparoscopic tissue manipulation. Fundamental laparoscopic surgery (FLS), on the other hand, only measures time and position accuracy. FLS is a commonly used training program for basic laparoscopic training through part tasks. The FLS is employed in most of the laparoscopic training systems, including box trainers and virtual reality (VR) simulators. However, many laparoscopic VR simulators lack force feedback and measure tissue damage solely through visual feedback based on virtual collisions. Few VR simulators that provide force feedback have subjective force metrics. To provide an objective force assessment for haptic skills training in the VR simulators, we extend the FLS part tasks to haptic-based FLS (HFLS), focusing on controlled force exertion. We interface the simulated HFLS part tasks with a customized bi-manual haptic simulator that offers five degrees of freedom (DOF) for force feedback. The proposed tasks are evaluated through face and content validity among laparoscopic surgeons of varying experience levels. The results show that trainees perform better in HFLS tasks. The average Likert score observed for face and content validity is greater than 4.6 ± 0.3 and 4 ± 0.5 for all the part tasks, which indicates the acceptance of the simulator among subjects for its appearance and functionality. Face and content validations show the need to improve haptic realism, which is also observed in existing simulators. To enhance the accuracy of force rendering, we incorporated a laparoscopic tool force model into the simulation. We study the effectiveness of the model through a psychophysical study that measures just noticeable difference (JND) for the laparoscopic gripping task. The study reveals an insignificant decrease in gripping-force JND. A simple linear model could be sufficient for gripper force feedback, and a non-linear LapTool force model does not affect the force perception for the force range of 0.5-2.5 N. Further study is required to understand the usability of the force model in laparoscopic training at a higher force range. Additionally, the construct validity of HFLS will confirm the applicability of the developed simulator to train surgeons with different levels of experience.

Asunto(s)

Artritis Reumatoide , Fibroblastos , Inmunomodulación , Sinoviocitos , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Humanos , Fibroblastos/inmunología , Sinoviocitos/inmunología , Sinoviocitos/metabolismo , Sinoviocitos/patología , Animales , Membrana Sinovial/inmunología , Membrana Sinovial/patología

RESUMEN

Osteoporosis is a widespread disease and affects over 500,000 people in Austria. Fragility fractures are associated with it and represent not only an individual problem for the patients, but also an enormous burden for the healthcare system. While trauma surgery care is well provided in Vienna, there is an enormous treatment gap in secondary prevention after osteoporotic fracture. Systematic approaches such as the Fracture Liaison Service (FLS) aim to identify patients with osteoporosis after fracture, to clarify diagnostically, to initiate specific therapy, and to check therapy adherence. The aim of this article is to describe the practical implementation and operational flow of an already established FLS in Vienna. This includes the identification of potential FLS inpatients, the diagnostic workup, and recommendations for an IT solution for baseline assessment and follow-up of FLS patients. We summarize the concept, benefits, and limitations of FLS and provide prospective as well as clinical and economic considerations for a city-wide FLS, managed from a central location. Future concepts of FLS should include artificial intelligence for vertebral fracture detection and simple IT tools for the implementation of FLS in the outpatient sector.

Asunto(s)

Fracturas Osteoporóticas , Prevención Secundaria , Humanos , Austria , Fracturas Osteoporóticas/economía , Fracturas Osteoporóticas/terapia , Prevención Secundaria/economía , Osteoporosis/terapia , Osteoporosis/economía , Osteoporosis/diagnóstico

RESUMEN

BACKGROUND: Rheumatoid arthritis (RA) is a chronic inflammatory disease induced by TNF-α, which increases fibroblast-like synoviocytes inflammation, resulting in cartilage destruction. The current work sought to comprehend the pathophysiological importance of TNF-α stimulation on differential protein expression and their regulation by apigenin using in-vitro and in-vivo models of RA. METHODS: The human RA synovial fibroblast cells were stimulated with or without TNF-α (10 ng/ml) and treated with 40 µM apigenin. In-silico, in-vitro and in-vivo studies were performed to confirm the pathophysiological significance of apigenin on pro-inflammatory cytokines and on differential expression of TTR and RAGE proteins. RESULTS: TNF-α induced inflammatory response in synoviocytes revealed higher levels of IL-6, IL-1ß, and TNF-α cytokines and upregulated differential expression of TTR and RAGE. In-silico results demonstrated that apigenin has a binding affinity towards TNF-α, indicating its potential effect in the inflammatory process. Both in-vitro and in-vivo results obtained by Western Blot analysis suggested that apigenin reduced the level of p65 (p = 0.005), TTR (p = 0.002), and RAGE (p = 0.020). CONCLUSION: The findings of this study suggested that TNF-α promotes the differential expression of pro-inflammatory cytokines, TTR, and RAGE via NF-kB pathways activation. Anti-inflammatory effect of apigenin impedes TNF-α mediated dysregulation or expression associated with RA pathogenesis.

Asunto(s)

Apigenina , Artritis Reumatoide , Receptor para Productos Finales de Glicación Avanzada , Factor de Necrosis Tumoral alfa , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Apigenina/farmacología , Humanos , Factor de Necrosis Tumoral alfa/metabolismo , Receptor para Productos Finales de Glicación Avanzada/metabolismo , Fibroblastos/metabolismo , Fibroblastos/efectos de los fármacos , Sinoviocitos/metabolismo , Sinoviocitos/efectos de los fármacos , Membrana Sinovial/metabolismo , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/patología , Citocinas/metabolismo , Animales , Inflamación/metabolismo , Inflamación/tratamiento farmacológico

RESUMEN

Introduction: Teaching in surgery must follow the evolution of medical techniques, procedures and technology. Since the 90s videosurgery has been developing and nowadays is the gold standard for many procedures in general surgery. As there are different characteristics from conventional surgery, residents need to practice in order to adapt to simulators. The world medical literature has established many training programs among them the Fundaments of laparoscopic Surgery (FLS). Evaluation of surgical technical abilities is a major tool for the residency programs. Objectives: To assess the effect of training acquiring skills of residents at first and second postgraduated level year (PGY1 and PGY2) before and after practice using low fidelity simulator based on tasks from FLS. Methods: A prospective study was conducted to determinate skills acquisition of residents at first and second postgraduated level year l (PGY1 and PGY2) before and after standardized practice using low fidelity simulator. Conclusions: There was a significant improve of the technical abilities reaching levels of improvement. Resident's performance was evident in the sample groups. First year residents (PGY1) achieved knowledge retention after training similar to the performance of second in the initial assessment, suggesting that the sequenced training carried out is equivalent to one year of practice in the general surgery residency program

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Introdução: O ensino em cirurgia deve acompanhar a evolução das técnicas, procedimentos e tecnologia. Desde a década de 90 a videocirurgia vem se desenvolvendo e hoje é a primeira escolha para muitos procedimentos em cirugia geral. Com características diferentes da cirurgia convencional, os residentes treinaram em simuladores para adaptação. A literatura médica mundial já apresenta programas consagrados para treinamento entre eles os Fundamentos da Cirurgia Laparoscópica (FLS). A avaliação das habilidades técnicas cirúrgicas é uma ferramenta importante em programas de residência médica. Objetivo: Avaliar o impacto do treinamento através de aquisição de habilidades dos médicos residentes em cirurgia geral antes e após treinamento teórico-prático em videocirurgia utilizando um simulador de baixa fidelidade baseado em exercícios do FLS. Método: Estudar de modo prospectivo a aquisição de habilidades de médicos residentes em cirurgia geral de primeiro e segundo ano (R1 e R2), antes e após realização de treinamento teórico-prático pré-definido. Conclusão: Houve melhora significativa com o treinamento atingindo níveis de conhecimento e aprimoramento semelhantes aos trabalhos da literatura mundial. A melhoria no desempenho dos residentes foi evidente em ambos os grupos da amostra. Aqueles do primeiro ano atingiram a concentração de conhecimento após o treinamento semelhante ao desempenho dos residentes de segundo ano na avaliação inicial, destacando que o treinamento sequenciado realizado equivale a um ano de prática no programa de residência geral de cirurgia

RESUMEN

Alzheimer's disease (AD) represents a pressing global health challenge, with an anticipated surge in diagnoses over the next two decades. This progressive neurodegenerative disorder unfolds gradually, with observable symptoms emerging after two decades of imperceptible brain changes. While traditional therapeutic approaches, such as medication and cognitive therapy, remain standard in AD management, their limitations prompt exploration into novel integrative therapeutic approaches. Recent advancements in AD research focus on entraining gamma waves through innovative methods, such as light flickering and electromagnetic fields (EMF) stimulation. Flickering light stimulation (FLS) at 40 Hz has demonstrated significant reductions in AD pathologies in both mice and humans, providing improved cognitive functioning. Additionally, recent experiments have demonstrated that APOE mutations in mouse models substantially reduce tau pathologies, with microglial modulation playing a crucial role. EMFs have also been demonstrated to modulate microglia. The exploration of EMFs as a therapeutic approach is gaining significance, as many recent studies have showcased their potential to influence microglial responses. Th article concludes by speculating on the future directions of AD research, emphasizing the importance of ongoing efforts in understanding the complexities of AD pathogenesis through a holistic approach and developing interventions that hold promise for improved patient outcomes.

RESUMEN

The S-locus lectin receptor kinases (G-LecRKs) have been suggested as receptors for microbe/damage-associated molecular patterns (MAMPs/DAMPs) and to be involved in the pathogen defense responses, but the functions of most G-LecRKs in biotic stress response have not been characterized. Here, we identified a member of this family, G-LecRK-I.2, that positively regulates flg22- and Pseudomonas syringae pv. tomato (Pst) DC3000-induced stomatal closure. G-LecRK-I.2 was rapidly phosphorylated under flg22 treatment and could interact with the FLS2/BAK1 complex. Two T-DNA insertion lines, glecrk-i.2-1 and glecrk-i.2-2, had lower levels of reactive oxygen species (ROS) and nitric oxide (NO) production in guard cells, as compared with the wild-type Col-0, under Pst DC3000 infection. Also, the immunity marker genes CBP60g and PR1 were induced at lower levels under Pst DC3000 hrcC- infection in glecrk-i.2-1 and glecrk-i.2-2. The GUS reporter system also revealed that G-LecRK-I.2 was expressed only in guard cells. We also found that G-LecRK-I.2 could interact H+-ATPase AHA1 to regulate H+-ATPase activity in the guard cells. Taken together, our results show that G-LecRK-I.2 plays an important role in regulating stomatal closure under flg22 and Pst DC3000 treatments and in ROS and NO signaling specifically in guard cells.

Asunto(s)

Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Receptores Mitogénicos/genética , Especies Reactivas de Oxígeno/metabolismo , ATPasas de Translocación de Protón/genética , Pseudomonas syringae/fisiología , Enfermedades de las Plantas/microbiología , Regulación de la Expresión Génica de las Plantas

RESUMEN

Trichloroethylene, a chlorinated solvent widely used as a degreasing agent, is a common environmental contaminant. Emerging evidence suggests that chronic exposure to trichloroethylene (TCE) contributes to the development of Parkinson's disease (PD). TCE induced LRRK2 kinase activity in the rat brain and produced a significant dopaminergic lesion in the nigrostriatal tract with elevated oxidative stress. Here we have utilized TCE-induced PD model for the assessment of test drug. Oral gavage administration of TCE at a dose of 1000 mg/kg/day for 6 weeks was utilized to induced PD. Muscle grip strength was estimated by rotarod and grid performance test. Motor activity by actophotometer and locomotor stability were assessed by forelimb locomotor scale (FLS) and forelimb step alternation test (FSAT). However, the postural stability was assessed by postural stability test (PST). Biochemical estimation consists of determination of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), GSH level (reduced glutathione), and nitrite concentration.

Asunto(s)

Enfermedad de Parkinson , Tricloroetileno , Ratas , Animales , Tricloroetileno/toxicidad , Ratas Wistar , Solventes , Estrés Oxidativo

RESUMEN

OBJECTIVE: Fibroblast-like synoviocytes (FLS) play a critical role on the exacerbation and deterioration of rheumatoid arthritis (RA). Aberrant activation of FLS pyroptosis signaling is responsible for the hyperplasia of synovium and destruction of cartilage of RA. This study investigated the screened traditional Chinese medicine berberine (BBR), an active alkaloid extracted from the Coptis chinensis plant, that regulates the pyroptosis of FLS and secretion of inflammatory factors in rheumatoid arthritis. METHODS: First, BBR was screened using a high-throughput drug screening strategy, and its inhibitory effect on RA-FLS was verified by in vivo and in vitro experiments. Second, BBR was intraperitoneally administrated into the collagen-induced arthritis rat model, and the clinical scores, arthritis index, and joint HE staining were evaluated. Third, synovial tissues of CIA mice were collected, and the expression of NLRP3, cleaved-caspase-1, GSDMD-N, Mst1, and YAP was detected by Western blot. RESULTS: The administration of BBR dramatically alleviated the severity of collagen-induced arthritis rat model with a decreased clinical score and inflammation reduction. In addition, BBR intervention significantly attenuates several pro-inflammatory cytokines (interleukin-1ß, interleukin-6, interleukin-17, and interleukin-18). Moreover, BBR can reduce the pyroptosis response (caspase-1, NLR family pyrin domain containing 3, and gasdermin D) of the RA-FLS in vitro, activating the Hippo signaling pathway (Mammalian sterile 20-like kinase 1, yes-associated protein, and transcriptional enhanced associate domains) so as to inhibit the pro-inflammatory effect of RA-FLS. CONCLUSION: These results support the role of BBR in RA and may have therapeutic implications by directly repressing the activation, migration of RA-FLS, which contributing to the attenuation of the progress of CIA. Therefore, targeting PU.1 might be a potential therapeutic approach for RA. Besides, BBR inhibited RA-FLS pyroptosis by downregulating of NLRP3 inflammasomes (NLRP3, caspase-1) and eased the pro-inflammatory activities via activating the Hippo signaling pathway, thereby improving the symptom of CIA.

Asunto(s)

Artritis Experimental , Artritis Reumatoide , Berberina , Ratas , Ratones , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Berberina/farmacología , Berberina/uso terapéutico , Berberina/metabolismo , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Membrana Sinovial/metabolismo , Caspasas/metabolismo , Caspasas/farmacología , Caspasas/uso terapéutico , Fibroblastos/metabolismo , Células Cultivadas , Proliferación Celular , Mamíferos

RESUMEN

The pathogenesis of osteoarthritis (OA) is still unclear, leading to the lack of targeted treatment. We aimed to probe into the effect of apolipoprotein D (APOD), the key gene from our previous study through bioinformatics analysis, on fibroblast-like synoviocyte (FLS) and chondrocytes in vitro to confirm its potential roles on the delay of OA progression. Primary FLS and chondrocytes were extracted from synovium and cartilage of OA patients and stimulated with interleukin 1ß (IL-1ß) in vitro. After APOD intervention, viability and proliferation of FLS and chondrocytes were detected. Subsequently, the inflammatory factors of the two cells were detected by quantitative reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and western blot, and the apoptosis and autophagy-related substances were determined at the same time. Finally, the oxidation level in FLS and chondrocytes were detected. APOD reversed the change of gene expression stimulated by IL-1ß in FLS and chondrocytes. APOD alleviated the proliferation of FLS while promoted proliferation of chondrocytes, and reduced the expression of inflammatory factors. Moreover, APOD promoted apoptosis of FLS and autography of chondrocytes, while reduced apoptosis of chondrocytes. Finally, decrease level of reactive oxygen species (ROS) in both cells were observed after APOD intervention, as well as the increased expression of antioxidant-related genes. APOD had effects on the proliferation of FLS and chondrocytes through apoptosis and autography as well as the reduction of oxidative stress, delaying the progress of OA.

Asunto(s)

MicroARNs , Osteoartritis , Sinoviocitos , Humanos , Sinoviocitos/metabolismo , Condrocitos/metabolismo , Apolipoproteínas D/metabolismo , Osteoartritis/metabolismo , Interleucina-1beta/metabolismo , Fibroblastos/patología , Apoptosis , MicroARNs/metabolismo

RESUMEN

Notch and its ligands play a critical role in rheumatoid arthritis (RA) pathogenesis. Hence, studies were conducted to delineate the functional significance of the Notch pathway in RA synovial tissue (ST) cells and the influence of RA therapies on their expression. Morphological studies reveal that JAG1, DLL4, and Notch1 are highly enriched in RA ST lining and sublining CD68+CD14+ MΦs. JAG1 and DLL4 transcription is jointly upregulated in RA MΦs reprogrammed by TLR4/5 ligation and TNF, whereas Syntenin-1 exposure expands JAG1, DLL4, and Notch1 expression levels in these cells. Single-cell RNA-seq data exhibit that JAG1 and Notch3 are overexpressed on all fibroblast-like synoviocyte (FLS) subpopulations, in parallel, JAG2, DLL1, and Notch1 expression levels are modest on RA FLS and are predominately potentiated by TLR4 ligation. Intriguingly, JAG1, DLL1/4, and Notch1/3 are presented on RA endothelial cells, and their expression is mutually reconfigured by TLR4/5 ligation in the endothelium. Synovial JAG1/JAG2/DLL1 or Notch1/3 transcriptomes were unchanged in patients who received disease-modifying anti-rheumatic drugs (DMARDs) or IL-6R Ab therapy regardless of disease activity score. Uniquely, RA MΦs and endothelial cells rewired by IL-6 displayed DLL4 transcriptional upregulation, and IL-6R antibody treatment disrupted RA ST DLL4 transcription in good responders compared to non-responders or moderate responders. Nevertheless, the JAG1/JAG2/DLL1/DLL4 transcriptome was diminished in anti-TNF good responders with myeloid pathotype and was unaltered in the fibroid pathotype except for DLL4. Taken together, our findings suggest that RA myeloid Notch ligands can serve as markers for anti-TNF responsiveness and trans-activate Notch receptors expressed on RA FLS and/or endothelial cells.

Asunto(s)

Artritis Reumatoide , Inhibidores del Factor de Necrosis Tumoral , Humanos , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Proteínas de Unión al Calcio/metabolismo , Proteínas de la Membrana/metabolismo , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Células Endoteliales/metabolismo , Receptor Toll-Like 4/metabolismo , Receptores Notch/metabolismo , Biomarcadores , Artritis Reumatoide/tratamiento farmacológico , Ligandos , Receptor Notch1/metabolismo

RESUMEN

OBJECTIVE: Fundamentals of Laparoscopic Surgery (FLS) is a multiple-choice test and a manual skills exam using simulation that Obstetrics and Gynecology (OBGYN) residents must pass to qualify for board certification. There is insufficient validity evidence supporting the use of FLS as a high-stakes exam. This study examines the correlation between OBGYN residents' performance on the FLS manual tasks and simulated vaginal cuff closure. METHODS: We compared residents' performance on FLS tasks with simulated vaginal cuff suturing on a model. During the first coached simulation session, after completion of training on the standard 5 FLS tasks, residents were coached on vaginal cuff closure using a simulated model placed inside the standard FLS box trainer. At a subsequent session, their performance was scored using the Global Operative Assessment of Laparoscopic Skills Scale (GOALS) and a second task-specific metric, and these scores were compared to their official FLS score. RESULTS: Twenty-nine residents completed the vaginal cuff simulation training between June 2019 and November 2021. Nineteen of the 29 were able to complete the cuff closure with the mean time to completion being 14.5 minutes. We found no correlation between official manual skills FLS scores and vaginal cuff GOALS scores (rho = -0.02, p = 0.90) or cuff closure assessment tool score (rho = -0.015, p = 0.048). There was also no correlation between time to completion for any FLS task and vaginal cuff closure OSAT scores. All residents reported that they found the cuff to be a useful addition to the FLS curriculum. CONCLUSIONS: Our study demonstrated that trainee performance on a simulated vaginal cuff closure model did not correlate with official FLS manual tasks skills. This finding adds to the body of evidence disputing the use of FLS as a high-stakes exam to assess laparoscopic skills in gynecology in the relationship with other variables category.

Asunto(s)

Ginecología , Internado y Residencia , Laparoscopía , Entrenamiento Simulado , Femenino , Embarazo , Humanos , Competencia Clínica , Ginecología/educación , Curriculum , Laparoscopía/educación

RESUMEN

Rheumatoid arthritis (RA) is a chronic, progressive, systemic autoimmune disease. Among them, abnormal proliferation, migration and vascularization of fibroblast-like synoviocytes (FLS) are the main pathological basis of persistent synovitis and bone destruction in RA. In the current study, we attempted to find effective molecular mechanisms for the treatment of RA by investigating RA-FLS. Firstly, the study was conducted to identify the potential target gene PTEN and its related signaling pathway through bioinformatics analysis. Subsequently, the target gene PTEN overexpression was regulated by cell transfection. The expression of FOXO signaling factors and autophagy-related proteins were detected by western blotting assay. Cell proliferation was measured by CCK-8 and EdU assays. Inflammation level was detected by ELISA. Cell migration and invasion were detected using wound healing assay and transwell chamber assay, respectively. Cell apoptosis was detected using flow cytometry. The results showed that overexpression of PTEN activated FOXO1 signaling in RA-FLS, and regulated autophagy, proliferation, invasion, migration, and the levels of pro-inflammatory factors in the disease. In conclusion, PTEN might provide an effective therapeutic strategy for rheumatoid arthritis by mediating the FOXO1 signaling pathway.

Asunto(s)

Artritis Reumatoide , Sinoviocitos , Humanos , Sinoviocitos/metabolismo , Artritis Reumatoide/metabolismo , Transducción de Señal , Inflamación/patología , Fibroblastos/metabolismo , Movimiento Celular , Proliferación Celular , Células Cultivadas , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/metabolismo

RESUMEN

The stability of the teleoperated cyber-physical system with model uncertainty, external disturbance, and actuator fault is addressed in this study by using a suitable fractional-order sliding mode control (SMC) strategy. First, the sliding surface is designed to ensure the better tracking performance of the system. Second, the suggested control method combines SMC with an adaptive strategy to ensure that the system is stable in finite time. Third, neural network (NN) and fuzzy logic system (FLS) are used to estimate the model uncertainty, time-varying delay, external disturbance and unknown coefficient matrices of sliding mode surface, respectively. Finally, the advantages of the proposed control scheme are confirmed by the simulation example.