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BACKGROUND: Post-traumatic capsular contracture is a common complication of joint injury and surgery. Post-traumatic capsular contracture is associated with fibrosis characterized by excessive differentiation and proliferation of myofibroblasts and abnormal secretion and accumulation of extracellular matrix. Previous studies have suggested that IL11 plays a role in myocardial fibrosis. We thus hypothesized that IL11 may play a fibrotic role during capsular contracture, in order to discover new targets for preventing joint capsule contracture METHODS: We constructed a post-traumatic contracture model by excessively extending the knee joint and fixing the joint in the flexion position, and a post-traumatic joint capsule contracture model was constructed in the wild-type, IL11-/-, IL11R -/-, α-SMA-cre-IL11fl/fl, α-SMA-cre-IL11Rfl/fl mouse strain, with wild-type mice without any treatment of the knee joint as the control group. Fibrotic markers and the expression of IL11 and IL11R in knee joint tissue were detected in each group of mice. The NIH3T3 cell line was used for in vitro analyses. The expression of fibrosis markers, IL11, TGFß and ERK1/2 were detected by western blot, ELISA and RT-qPCR. RESULTS: Inhibition of IL11 inhibited ERK1/2 phosphorylation, reduced the secretion of collagen in the joint capsule, and inhibited the excessive differentiation and proliferation of myofibroblasts in the post-traumatic joint capsule contracture, thus alleviating the joint capsule contracture and obtaining better joint mobility. CONCLUSION: Downregulation of IL11 in traumatic joint capsule contracture inhibits ERK1/2 phosphorylation, thus significantly relieving joint capsule contracture. Our findings indicate the TGFß/IL11/ERK1/2 axis is an important pathway for the differentiation of fibroblasts into myofibroblasts. Anti-IL11 treatment is an effective means to prevent traumatic joint capsule contracture.
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BACKGROUND: The traction table is generally used in femoral intramedullary nailing surgery. Recently, some published studies have shown that the same or better treatment effects can be gotten without a traction table. It remains no consensus on this issue. METHODS: The Preferred Reporting Items for Systematic Reviews and Meta-analyses guideline was applied in this study. We searched PubMed, Embase, Web of Science, and Cochrane Library databases for eligible studies. The random-effect model was used to calculate the standardized mean difference (SMD) and risk ratios with 95% CIs. Trial sequential analysis (TSA) was performed to verify the results. RESULTS: The pooled estimates of seven studies, including 266 cases each in the manual traction group and traction table group, indicated that manual traction could shorten operative time [SMD, - 0.77; 95% CI (- 0.98, - 0.55); P < 0.00001] and preoperative set-up time [SMD, - 2.37; 95% CI (- 3.90, - 0.84); P = 0.002], but it would not reduce intraoperative blood loss volume and fluoroscopy time. No statistical difference was found in their fracture healing time, postoperative Harris scores, and malunion rate. The use of a Traction repositor could reduce the set-up time [SMD, - 2.48; 95% CI (- 4.91, - 0.05); P < 0.00001]. CONCLUSIONS: Compared with manual traction, the traction table in femoral intramedullary nailing surgery lengthened operative time and preoperative set-up time. At the same time, it did not show significant advantages in reducing blood loss volume and fluoroscopy time, or improving prognosis. In clinical practice, the optimal surgical plan must be made on a case-by-case basis to avoid unnecessary traction table use.
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Fracturas del Fémur , Fijación Intramedular de Fracturas , Humanos , Fijación Intramedular de Fracturas/métodos , Tracción/métodos , Fracturas del Fémur/cirugía , Fémur , Curación de Fractura , Clavos Ortopédicos , Resultado del TratamientoRESUMEN
BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) had become the most prevalent liver disease worldwide. Early diagnosis could effectively reduce NAFLD-related morbidity and mortality. This study aimed to combine the risk factors to develop and validate a novel model for predicting NAFLD. METHODS: We enrolled 578 participants completing abdominal ultrasound into the training set. The least absolute shrinkage and selection operator (LASSO) regression combined with random forest (RF) was conducted to screen significant predictors for NAFLD risk. Five machine learning models including logistic regression (LR), RF, extreme gradient boosting (XGBoost), gradient boosting machine (GBM), and support vector machine (SVM) were developed. To further improve model performance, we conducted hyperparameter tuning with train function in Python package 'sklearn'. We included 131 participants completing magnetic resonance imaging into the testing set for external validation. RESULTS: There were 329 participants with NAFLD and 249 without in the training set, while 96 with NAFLD and 35 without were in the testing set. Visceral adiposity index, abdominal circumference, body mass index, alanine aminotransferase (ALT), ALT/AST (aspartate aminotransferase), age, high-density lipoprotein cholesterol (HDL-C) and elevated triglyceride (TG) were important predictors for NAFLD risk. The area under curve (AUC) of LR, RF, XGBoost, GBM, SVM were 0.915 [95% confidence interval (CI): 0.886-0.937], 0.907 (95% CI: 0.856-0.938), 0.928 (95% CI: 0.873-0.944), 0.924 (95% CI: 0.875-0.939), and 0.900 (95% CI: 0.883-0.913), respectively. XGBoost model presented the best predictive performance, and its AUC was enhanced to 0.938 (95% CI: 0.870-0.950) with further parameter tuning. CONCLUSIONS: This study developed and validated five novel machine learning models for NAFLD prediction, among which XGBoost presented the best performance and was considered a reliable reference for early identification of high-risk patients with NAFLD in clinical practice.
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Enfermedad del Hígado Graso no Alcohólico , Humanos , Enfermedad del Hígado Graso no Alcohólico/diagnóstico por imagen , Factores de Riesgo , Alanina Transaminasa , Área Bajo la Curva , Aprendizaje AutomáticoRESUMEN
The integrated stress response (ISR) is a homeostatic mechanism by which eukaryotic cells sense and respond to stress-inducing signals, such as amino acid starvation. General controlled non-repressed (GCN2) kinase is a key orchestrator of the ISR, and modulates protein synthesis in response to amino acid starvation. Here we demonstrate in mice that GCN2 controls intestinal inflammation by suppressing inflammasome activation. Enhanced activation of ISR was observed in intestinal antigen presenting cells (APCs) and epithelial cells during amino acid starvation, or intestinal inflammation. Genetic deletion of Gcn2 (also known as Eif2ka4) in CD11c(+) APCs or intestinal epithelial cells resulted in enhanced intestinal inflammation and T helper 17 cell (TH17) responses, owing to enhanced inflammasome activation and interleukin (IL)-1ß production. This was caused by reduced autophagy in Gcn2(-/-) intestinal APCs and epithelial cells, leading to increased reactive oxygen species (ROS), a potent activator of inflammasomes. Thus, conditional ablation of Atg5 or Atg7 in intestinal APCs resulted in enhanced ROS and TH17 responses. Furthermore, in vivo blockade of ROS and IL-1ß resulted in inhibition of TH17 responses and reduced inflammation in Gcn2(-/-) mice. Importantly, acute amino acid starvation suppressed intestinal inflammation via a mechanism dependent on GCN2. These results reveal a mechanism that couples amino acid sensing with control of intestinal inflammation via GCN2.
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Aminoácidos/metabolismo , Colitis/metabolismo , Inflamasomas/antagonistas & inhibidores , Inflamación/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/patología , Proteínas Serina-Treonina Quinasas/metabolismo , Aminoácidos/administración & dosificación , Aminoácidos/deficiencia , Aminoácidos/farmacología , Animales , Células Presentadoras de Antígenos/inmunología , Células Presentadoras de Antígenos/metabolismo , Autofagia , Proteína 5 Relacionada con la Autofagia , Proteína 7 Relacionada con la Autofagia , Colitis/etiología , Colitis/patología , Colitis/prevención & control , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Femenino , Humanos , Inflamasomas/metabolismo , Inflamación/etiología , Inflamación/patología , Inflamación/prevención & control , Interleucina-1beta/inmunología , Masculino , Ratones , Proteínas Asociadas a Microtúbulos/deficiencia , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas Serina-Treonina Quinasas/deficiencia , Proteínas Serina-Treonina Quinasas/genética , Especies Reactivas de Oxígeno/metabolismo , Estrés Fisiológico , Células Th17/inmunología , Enzimas Activadoras de Ubiquitina/deficiencia , Enzimas Activadoras de Ubiquitina/metabolismoRESUMEN
Ellagic acid has been shown to inhibit tumor cell growth. However, the underlying molecular mechanisms remain elusive. In this study, our aim was to investigate whether ellagic acid inhibits the proliferation of MCF- 7 human breast cancer cells via regulation of the TGF-ß/Smad3 signaling pathway. MCF-7 breast cancer cells were transfected with pEGFP-C3 or pEGFP-C3/Smad3 plasmids, and treated with ellagic acid alone or in combination with SIS3, a specific inhibitor of Smad3 phosphorylation. Cell proliferation was assessed by MTT assay and the cell cycle was detected by flow cytometry. Moreover, gene expression was detected by RT-PCR, real-time PCR and Western blot analysis. The MTT assay showed that SIS3 attenuated the inhibitory activity of ellagic acid on the proliferation of MCF-7 cells. Flow cytometry revealed that ellagic acid induced G0/G1 cell cycle arrest which was mitigated by SIS3. Moreover, SIS3 reversed the effects of ellagic acid on the expression of downstream targets of the TGF-ß/Smad3 pathway. In conclusion, ellagic acid leads to decreased phosphorylation of RB proteins mainly through modulation of the TGF-ß/Smad3 pathway, and thereby inhibits the proliferation of MCF-7 breast cancer cells.
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Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Ácido Elágico/farmacología , Transducción de Señal/efectos de los fármacos , Proteína smad3/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Expresión Génica/efectos de los fármacos , Humanos , Isoquinolinas/farmacología , Células MCF-7 , Piridinas/farmacología , Pirroles/farmacología , Proteína de Retinoblastoma/genética , Proteína de Retinoblastoma/metabolismo , Proteína smad3/antagonistas & inhibidores , Proteína smad3/genéticaRESUMEN
The root of Polygonum multiflorum Thunb. (PM) is utilized to treat many diseases associated with aging. Research also indicates that PM inhibits the proliferation of certain types of cancer cells. The aim of the present study was to evaluate the inhibitory effect of PM extract (PME) on the proliferation of MCF-7 cells and to investigate the underlying mechanisms. Inhibition of the proliferation of MCF-7 cells was determined by the MTT assay. Cell cycle distribution and apoptotic rates were evaluated by flow cytometry, and cell cycle and apoptosis-related protein expression was assessed by Western blotting. Apoptotic characteristics of MCF-7 cells were detected by transmission electron microscopy. The present study showed that PME at doses of 100, 150, 200 and 250 µg/ml signiï¬cantly inhibited proliferation of MCF-7 cells in a time- and dose-dependent manner. Flow cytometry showed that the cell apoptotic rates were 9.1 ± 1.67 and 17.7 ± 2.93% after treatment with 100 and 200 µg/ml PME for 48 h, respectively. The proportions of cells in the G2/M phase were 37.9 ± 1.47 and 42.0 ± 1.71% after treatment with 100 and 200 µg/ml PME for 24 h, respectively. Western blot analysis showed that PME down-regulated the protein expression of Cdc25B and Cdc25C phosphatases accompanied by an increase in phospho-Cdk1, and PME promoted cytochrome c release from mitochondria into the cytosol to activate caspase-9. The present study demonstrated that PME inhibited MCF-7 cell proliferation by inducing cell cycle arrest in the G2/M phase and promoting cell apoptosis. The effects of PME on MCF-7 cells were associated with the modulation of the expression levels of proteins involved in the cell cycle and apoptosis. These data suggest that PME has promise as a treatment against breast cancer by inhibiting the proliferation of cancer cells.
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Antineoplásicos Fitogénicos/farmacología , Neoplasias de la Mama/metabolismo , Extractos Vegetales/farmacología , Polygonum/química , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/uso terapéutico , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Proteína Quinasa CDC2/metabolismo , Caspasa 9/metabolismo , Puntos de Control del Ciclo Celular , Línea Celular Tumoral , Citocromos c/metabolismo , Femenino , Citometría de Flujo , Humanos , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Raíces de Plantas/química , Fosfatasas cdc25/metabolismoRESUMEN
Dendritic cells (DCs) play a vital role in initiating robust immunity against pathogens as well as maintaining immunological tolerance to self antigens. However, the intracellular signaling networks that program DCs to become tolerogenic remain unknown. We report here that the Wnt-beta-catenin signaling in intestinal dendritic cells regulates the balance between inflammatory versus regulatory responses in the gut. beta-catenin in intestinal dendritic cells was required for the expression of anti-inflammatory mediators such as retinoic acid-metabolizing enzymes, interleukin-10, and transforming growth factor-beta, and the stimulation of regulatory T cell induction while suppressing inflammatory effector T cells. Furthermore, ablation of beta-catenin expression in DCs enhanced inflammatory responses and disease in a mouse model of inflammatory bowel disease. Thus, beta-catenin signaling programs DCs to a tolerogenic state, limiting the inflammatory response.
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Células Dendríticas/inmunología , Inflamación , Enfermedades Inflamatorias del Intestino/inmunología , Mucosa Intestinal/inmunología , Autotolerancia , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunología , beta Catenina/metabolismo , Animales , Citocinas/metabolismo , Células Dendríticas/metabolismo , Perfilación de la Expresión Génica , Mucosa Intestinal/citología , Mucosa Intestinal/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Transducción de Señal , Linfocitos T Colaboradores-Inductores/citología , Tretinoina/metabolismo , Proteínas Wnt/metabolismoRESUMEN
In vitro measures of immune responsiveness toward tumors provide relevant information regarding the prevention and metastatic potential of cancer. In addition, the compartmentalization of immune responses is likely to be an important factor in dictating host antitumor immune responses. We have previously demonstrated that injection of antibody against B cells diminished pulmonary antitumor defenses. In the current study, we determined the effect of B cells on antitumor cellular responses against a lung metastatic tumor, MADB106. Lung B cells displayed sustained surface expression of CD80 and CD86, as compared to spleen B cells, in the presence of MADB106 tumor. Removal of B cells from lung lymphocyte cultures resulted in diminished IFN-gamma secretion and tumor lysis, whereas removal of B cells from spleen lymphocytes exposed to tumor resulted in elevated IFN-gamma and increased tumor lysis. Furthermore, a correlative increase in CD80 and CD86 co-stimulatory molecule expression by lung B cells was observed in mice subjected to MADB106 tumor. These findings provide additional evidence of the importance of pulmonary B cell responses in tumor defenses.
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Adenocarcinoma/secundario , Linfocitos B/inmunología , Vigilancia Inmunológica , Neoplasias Pulmonares/inmunología , Neoplasias Experimentales/inmunología , Bazo/inmunología , Adenocarcinoma/inmunología , Adenocarcinoma/patología , Animales , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Interferón gamma/metabolismo , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Neoplasias Pulmonares/patología , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos Infiltrantes de Tumor/patología , Masculino , Neoplasias Experimentales/patología , Ratas , Ratas Endogámicas F344RESUMEN
The intestinal immune system must elicit robust immunity against harmful pathogens but must also restrain immune responses directed against commensal microbes and dietary antigens. The mechanisms that maintain this dichotomy are poorly understood. Here we describe a population of CD11b+F4/80+CD11c- macrophages in the lamina propria that expressed several anti-inflammatory molecules, including interleukin 10 (IL-10), but little or no proinflammatory cytokines, even after stimulation with Toll-like receptor ligands. These macrophages induced, by a mechanism dependent on IL-10, retinoic acid and exogenous transforming growth factor-beta, the differentiation of Foxp3+ regulatory T cells. In contrast, lamina propria CD11b+ dendritic cells elicited IL-17 production. This IL-17 production was suppressed by lamina propria macrophages, indicating that a dynamic interaction between these subsets may influence the balance between immune activation and tolerance.
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Células Dendríticas/fisiología , Interleucina-17/biosíntesis , Intestinos/inmunología , Macrófagos/fisiología , Membrana Mucosa/citología , Linfocitos T Reguladores/inmunología , Animales , Antígeno CD11b/análisis , Linfocitos T CD4-Positivos/inmunología , Diferenciación Celular , Tolerancia Inmunológica , Masculino , Ratones , Ratones Endogámicos C57BL , Factor de Crecimiento Transformador beta/fisiologíaRESUMEN
Optimal activation of naïve T lymphocyte requires two signals; an antigen-specific signal initiated by engagement of TCR with the antigen-MHC complex and a costimulatory signal independent of the antigen receptor complex. Without the costimulatory signal, T cells become anergic. Various adhesion molecules, such as B7-1 (CD80) and heat stable antigen (HSA, CD24), expressed on antigen presenting cells have been demonstrated to provide costimulatory signals to T cells. It was reported that the combinations of different adhesion molecules could induce even stronger immune response. In this study, we made a hybrid costimulatory molecule, B7-1-HSA, and tested its T cell stimulatory function. Chinese hamster ovary (CHO) cells expressing this hybrid molecule bound both anti-CD80 and anti-CD24 monoclonal antibodies, and induced stronger T cell proliferation than CHO cells expressing B7-1 or HSA alone. These results suggest that the B7-1-HSA hybrid molecule can deliver two costimulatory signals simultaneously that can synergize in inducing T cell proliferation. The purified B7-1-HSA protein reacted with both anti-B7-1 and anti-HSA mAbs in Western blotting and specifically mediated adhesion of Jurkat cells. Furthermore, purified B7-1-HSA molecule spontaneously incorporated onto cell membrane through its glycolipid anchor suggesting that this hybrid costimulatory molecule can be used in protein transfer to develop effective cancer vaccines.