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IκB kinase (IKK) complex is central regulators of the NF-κB pathway, and dysregulation of IKK phosphorylation leads to hyperactivation of proinflammatory response in various chronic inflammatory diseases, including inflammatory bowel disease (IBD). However, the dynamic modulation of IKK phosphorylation and dephosphorylation in intestinal inflammation remains uncharacterized. Here, we found that autophagy/beclin-1 regulator 1 (AMBRA1) was highly expressed in inflamed colons in a colitis mouse model and in clinical IBD samples. Importantly, AMBRA1 deletion significantly decreased proinflammatory cytokine expression and enhanced the therapeutic effect of infliximab on intestinal inflammation. Mechanistically, the N-term F1 domain of AMBRA1 was required for AMBRA1 to competitively interact with protein phosphatase 4 regulatory subunit 1 (PP4R1) and catalytic protein phosphatase 4 (PP4c) to suppress their interactions with IKK, promote the dissociation of the PP4R1/PP4c complex, and antagonize the dephosphorylation activity of this complex towards the IKK complex. In response to TNF-α stimulation, IKKα phosphorylates AMBRA1 at S1043 to stabilize AMBRA1 expression by impairing its binding to Cullin4A (CUL4A) to decrease its CUL4A-mediated K48-linked ubiquitination. Overall, our study identifies an autophagy-independent function of AMBRA1 as a positive modulator of IKK phosphorylation to promote intestinal inflammation, thus providing a new targeted therapeutic strategy for patients with refractory IBD.
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Proteínas Adaptadoras Transductoras de Señales , Autofagia , Quinasa I-kappa B , Animales , Humanos , Ratones , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Autofagia/efectos de los fármacos , Colitis/metabolismo , Colitis/patología , Colitis/inducido químicamente , Células HEK293 , Quinasa I-kappa B/metabolismo , Inflamación/metabolismo , Inflamación/patología , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/patología , Ratones Endogámicos C57BL , Fosfoproteínas Fosfatasas/metabolismo , Fosfoproteínas Fosfatasas/genética , FosforilaciónRESUMEN
Introduction: Over the past few years, multiple observational studies have speculated a potential association between inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn's disease (CD), and osteoporosis. However, no consensus has been reached regarding their interdependence and pathogenesis. Herein, we sought to further explore the causal associations between them. Methods: We validated the association between IBD and reduced bone mineral density in humans based on genome-wide association studies (GWAS) data. To investigate the causal relationship between IBD and osteoporosis, we performed a two-sample Mendelian randomization study using training and validation sets. Genetic variation data for IBD, CD, UC, and osteoporosis were derived from published genome-wide association studies in individuals of European ancestry. After a series of robust quality control steps, we included eligible instrumental variables (SNPs) significantly associated with exposure (IBD/CD/UC). We adopted five algorithms, including MR Egger, Weighted median, Inverse variance weighted, Simple mode, and Weighted mode, to infer the causal association between IBD and osteoporosis. In addition, we evaluated the robustness of Mendelian randomization analysis by heterogeneity test, pleiotropy test, leave-one-out sensitivity test, and multivariate Mendelian randomization. Results: Genetically predicted CD was positively associated with osteoporosis risk, with ORs of 1.060 (95% CIs 1.016, 1.106; p = 0.007) and 1.044 (95% CIs 1.002, 1.088; p = 0.039) for CD in the training and validation sets, respectively. However, Mendelian randomization analysis did not reveal a significant causal relationship between UC and osteoporosis (p > 0.05). Furthermore, we found that overall IBD was associated with osteoporosis prediction, with ORs of 1.050 (95% CIs 0.999, 1.103; p = 0.055) and 1.063 (95% CIs 1.019, 1.109; p = 0.005) in the training and validation sets, respectively. Conclusion: We demonstrated the causal association between CD and osteoporosis, complementing the framework for genetic variants that predispose to autoimmune disease.
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Colitis Ulcerosa , Enfermedades Inflamatorias del Intestino , Osteoporosis , Humanos , Análisis de la Aleatorización Mendeliana , Estudio de Asociación del Genoma Completo , Enfermedades Inflamatorias del Intestino/complicaciones , Enfermedades Inflamatorias del Intestino/genética , Colitis Ulcerosa/complicaciones , Colitis Ulcerosa/genética , Osteoporosis/epidemiología , Osteoporosis/genéticaRESUMEN
Crohn's disease is a highly heterogeneous autoimmune disease with a unique inflammatory phenotype of T cells at the lesion site. We aim to further explore the diagnosis of Crohn's disease and drug prediction of T cell marker gene expression. We obtained single-cell expression profile data from 22 CDs or normal samples and performed cell annotation and cellular communication analysis. Through the intersection of T cell marker genes, differential genes, and WGCNA results, we identified T cell-specific key genes and their immune landscapes and potential pathogenesis, and validated them across multiple datasets and patient tissue samples. We also explored the differentiation characteristics of genes by pseudo-temporal analysis and assessed their diagnostic performance and drug sensitivity by molecular docking. Finally, we extended this study to the prognosis of IBD-associated colon cancer. TNF-centered 5-gene diagnostic model not only has excellent diagnostic efficacy, but is also closely associated with KRAS, P53, and IL6/JAK/STAT3 pathways and physiological processes, such as EMT, coagulation, and apoptosis. In addition, this diagnostic model may have potential synergistic immunotherapeutic effects, with positive correlations with immune checkpoints such as CTLA4, CD86, PDCD1LG2, and CD40. Molecular docking demonstrated that BIRC3 and ANXA1 have strong binding properties to Azathioprine and Glucoocorticoid. Furthermore, the 5-gene model may suggest antagonism to IFX and prognosis for colon cancer associated with inflammatory bowel disease. Single-cell sequencing targeting T cell-related features in patients with Crohn's disease may aid in new diagnostic decisions, as well as the initial exploration of high-potential therapies.
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Neoplasias del Colon , Enfermedad de Crohn , Enfermedades Inflamatorias del Intestino , Humanos , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/tratamiento farmacológico , Enfermedad de Crohn/genética , Simulación del Acoplamiento Molecular , Enfermedades Inflamatorias del Intestino/metabolismo , Linfocitos T/metabolismo , Neoplasias del Colon/metabolismoRESUMEN
Herein, nickel-aluminum-layered double hydroxide (NiAl-LDH) films were prepared by the hydrothermal method. Based on the photoinduced reduction ability and degradation of LDHs on heavy metal ions and organic compounds, NiAl-LDH films displayed favorable simultaneous removal performance. Benefiting from the electron traps of heavy metals reduced from solution, the coexisting metal ions improved the photocatalytic activity of NiAl-LDH films on methyl orange. The higher the Fermi level of coexisting metal ion was, the higher the photocatalytic degradation rate of methyl orange obtained. Meanwhile, the removal rates of heavy metal ions (Ag+, Pb2+, and Cu2+) from wastewater were both enhanced and could reach 95%. NiAl-LDH films showed affinity toward Ag+. Furthermore, NiAl-LDH films are tightly coupled with the substrate, providing active sites and a simple method for the catalyst recovery. This study provides new insights into the simultaneous removal of heavy metal ions and organic pollutants using LDH films.
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Interleukin (IL)-35 has capacity to ameliorate experimental colitis and is upregulated in immune cells from active Crohn's disease (CD) patients. Nonetheless, CD continues to be an incurable disease with characteristics of chronic relapsing-remitting inflammation. In this study, we investigated the production of IL-35 by B cells from active CD patients and non-CD controls. Immediately ex vivo, the mRNA transcription of IL-12/IL-35 subunit IL12A was significantly higher in CD B cells than in control B cells, and the mRNA transcription of IL-27/IL-35 subunit EBI3 was significantly higher in mild CD and moderate CD B cells than in control B cells. However, we also found that CpG-activated B cells and BCR + CD40-activated B cells from CD subjects presented significantly lower IL12A and lower EBI3 transcription than their counterparts from control subjects. We further evaluated IL-35 protein secretion and confirmed that B cell-mediated IL-35 protein secretion was lower in CD patients than in controls. However, IL-35-Fc preconditioning was able to significantly increase IL-35 production from B cells and to eliminate the difference in IL-35 production capacity between controls and CD patients. Furthermore, these IL-35-Fc-preconditioned B cells were able to suppress IFN-γ and IL-17 production from CD4+CD25- T cells more potently than Fc control-preconditioned B cells. Rh IL-27Rα-Fc, a soluble form of B cell-specific IL-35 receptor, significantly increased IFN-γ and IL-17 production. Together, these data supported a role of B cell-mediated IL-35 in suppressing IFN-γ and IL-17 inflammation. However, despite the fact that CD B cells presented higher transcription of IL-35 subunits directly ex vivo, CD B cells also presented reduced capacity for further IL-35 production upon activation.
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Linfocitos B/inmunología , Enfermedad de Crohn/inmunología , Interleucinas/inmunología , Adulto , Linfocitos T CD4-Positivos/inmunología , Células Cultivadas , Colitis/inmunología , Femenino , Humanos , Inflamación/inmunología , Interferón gamma/inmunología , Leucocitos Mononucleares/inmunología , Activación de Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Transcripción Genética/inmunología , Adulto JovenRESUMEN
Pathogenic inflammation mediated by overactive type 1 helper T cell (Th1) responses could exacerbate and perpetuate Crohn's disease. Programmed death (PD)-1 and its ligand PD-L1 pathway could be upregulated to suppress inflammation. We wondered why this pathway is ineffective at suppressing pathogenic Th1 inflammation in Crohn's disease patients. Here, we found that overexpression of T-bet via transfection significantly reduced the expression of PD-1. PD-L1 was capable of suppression proinflammatory CD4+ T cells, but T-bet transfection significantly reduced the susceptibility of CD4+ T cells toward PD-L1-mediated suppression, evidenced by the observations that at low PD-L1 concentration T-bet transfected and mock transfected CD4+ T cells presented comparable IL-2 production, but at high PD-L1 concentration, T-bet transfected CD4+ T cells presented significantly higher IL-2 than mock transfected CD4+ T cells. PD-L1 could significantly reduce the survival of CD4+ T cells from Crohn's disease patients, but interestingly, in the absence of PD-L1, the survival was better in mock transfected CD4+ T cells, while in the presence of PD-L1, the survival was better in T-bet transfected CD4+ T cells. Crohn's disease patients with greater severity presented higher T-bet expression and lower PD-1 expression in CD4+ T cells, demonstrating an association between T-bet expression and disease progression. We also discovered that stimulation with bacterial antigens could upregulate the expression of T-bet. Together, this study demonstrated that T-bet overexpression could interfere with PD-1/PD-L1-mediated suppression of CD4+ T cell inflammation and survival, and potentially contributed to the development and persistence of Crohn's disease.
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Antígeno B7-H1/genética , Linfocitos T CD4-Positivos/patología , Enfermedad de Crohn/genética , Enfermedad de Crohn/inmunología , Regulación de la Expresión Génica , Receptor de Muerte Celular Programada 1/genética , Proteínas de Dominio T Box/metabolismo , Adulto , Estudios de Casos y Controles , Supervivencia Celular , Enfermedad de Crohn/metabolismo , Humanos , Inflamación/metabolismoRESUMEN
Due to multiple charge transport pathways, adjustable layer spacing, compositional flexibility, low manufacturing cost, and absorption of visible light, layered double hydroxides (LDHs) are a promising material for wastewater treatment. In this study, LDH films and Fe-doped LDH films with different metal ions (Ni, Al, Fe) on the surface of conductive cloth were successfully prepared and applied for the photocatalytic degradation of wastewater containing methyl orange and Ag ions under visible-light irradiation. The chemical state of Fe ions and the composition of LDHs on methyl orange photodegradation were investigated. The experimental results showed that LDH films exhibited high photocatalytic activity. The photocatalytic activity of LDH films on methyl orange improved in the mixed wastewater, and the Fe-doped NiAl-LDH films exhibited best visible-light photocatalytic performance. The analysis showed that Ag ions in the mixed wastewater were reduced by the LDH films and subsequently deposited on the surface of the LDH films. The Ag nanoparticles acted as electron traps and promoted the photocatalytic activity of the LDH films on methyl orange. Thus, we have demonstrated that prepared LDH films can be used in the treatment of mixed wastewater and have broad application prospects in environmental remediation and purification processes.
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Crohn's disease (CD) is characterized by a pathogenic intestinal inflammation mediated by a Th1-skewed immune system and a dysregulated intestinal mucosal community. In this study, we investigated the role of bacteria on the activation and function of monocytes, and its association with CD pathogenesis. To this end, fecal bacteria from CD patients and healthy controls were collected and used to stimulate autologous circulating monocytes. Fecal bacteria from CD patients were more effective at upregulating NOD2, NLRP3, TLR2, and TLR4 expression than fecal bacteria from healthy controls. Furthermore, the monocyte-derived macrophages (MDMs) induced by CD bacteria were more sensitive to E. coli stimulation than the MDMs induced by control bacteria, and demonstrated more M1 characteristics with high IL-6, TNF-α, and IL-12 and low IL-4 production. These effects mediated by fecal bacteria from CD patients could be repressed by the supplementation of IL-4. IL-4 not only suppressed the expression of NOD1, NOD2, NLRP3, TLR2, TLR4, and CD14 in MDMs induced by CD bacteria, but also suppressed E. coli-mediated expression of IL-6, TNF-α, and IL-12. Furthermore, IL-4-conditioned MDMs were more effective at supporting Th2 differentiation and inhibiting Th1 and Th17 differentiation of CD4+ T cells. Together, these studies demonstrated that fecal bacteria from CD patients presented enhanced capacity to upregulate pattern-recognition molecules in macrophages, which could be repressed by IL-4.
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Enfermedad de Crohn/microbiología , Heces/microbiología , Interleucina-4/metabolismo , Adulto , Estudios de Casos y Controles , Diferenciación Celular , Enfermedad de Crohn/inmunología , Escherichia coli/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Interleucina-12/genética , Interleucina-12/metabolismo , Interleucina-4/genética , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Monocitos/metabolismo , Monocitos/microbiología , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteína Adaptadora de Señalización NOD1/genética , Proteína Adaptadora de Señalización NOD1/metabolismo , Proteína Adaptadora de Señalización NOD2/genética , Proteína Adaptadora de Señalización NOD2/metabolismo , Células TH1/citología , Células Th17/citología , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Secreted frizzled-related protein 1 (SFRP1) is a member of the SFRPs family that modulates the Wnt signal transduction pathway. Recent studies have showed down-regulation of SFRP1 expression in colorectal cancer (CRC). We aimed to evaluate the effect of SFRP1 on the proliferation, migration, invasion and apoptosis of CRC cells in vitro. MATERIALS AND METHODS: We used real-time fluorescence quantification (RT-PCR) and Western blotting to detect SFRP1 expression in CRC, pericarcinomatous tissues and CRC cell lines. We assessed the influence of overexpression and knockdown of SFRP1 on CRC cell proliferation, migration, invasion, and apoptosis, Western blotting was used to evaluate protein levels of Wnt, ß-catenin, and apoptosis-related proteins. RESULTS: The expression of SFRP1 was significantly decreased in CRC tissues. Among the six CRC cell lines (sw-480, sw1116, caco-2, ht-29, colo-205, and hct-116), RT-PCR revealed that sw1116 cells had the lowest expression of SFRP1, while caco-2 cells had the highest SFRP1 expression. SFRP1 overexpression in sw1116 cells significantly suppressed cell proliferation while SFRP1 knockdown in caco-2 cells significantly increase the cell proliferation. In addition, overexpression of SFRP1 in sw1116 cells remarkedly suppressed cell migration and invasion, whereas knockdown of SFRP1 in caco-2 cells resulted in significant enhancement of migration and invasion. Furthermore, SFRP1 overexpression in sw1116 cells promoted cell apoptosis. Western blotting showed that SFRP1 overexpression significantly decreased the protein levels of Wnt, ß-catenin and apoptosis-related proteins, including MMP2, MMP9, Twist, CDK1, TGF, and Bcl2. CONCLUSION: Our results demonstrate that SFRP1 suppresses cell proliferation, migration and invasion, and promotes apoptosis in CRC cells.
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Colorectal cancer (CRC) is one of the most common cancers and a major cause of mortality. The present study aimed to identify potential biomarkers for CRC metastasis and uncover the mechanisms underlying the etiology of the disease. The five datasets GSE68468, GSE62321, GSE22834, GSE14297 and GSE6988 were utilized in the study, all of which contained metastatic and non-metastatic CRC samples. Among them, three datasets were integrated via meta-analysis to identify the differentially expressed genes (DEGs) between the two types of samples. A protein-protein interaction (PPI) network was constructed for these DEGs. Candidate genes were then selected by the support vector machine (SVM) classifier based on the betweenness centrality (BC) algorithm. A CRC dataset from The Cancer Genome Atlas database was used to evaluate the accuracy of the SVM classifier. Pathway enrichment analysis was carried out for the SVM-classified gene signatures. In total, 358 DEGs were identified by metaanalysis. The top ten nodes in the PPI network with the highest BC values were selected, including cAMP responsive element binding protein 1 (CREB1), cullin 7 (CUL7) and signal sequence receptor 3 (SSR3). The optimal SVM classification model was established, which was able to precisely distinguish between the metastatic and non-metastatic samples. Based on this SVM classifier, 40 signature genes were identified, which were mainly enriched in protein processing in endoplasmic reticulum (e.g., SSR3), AMPK signaling pathway (e.g., CREB1) and ubiquitin mediated proteolysis (e.g., FBXO2, CUL7 and UBE2D3) pathways. In conclusion, the SVM-classified genes, including CREB1, CUL7 and SSR3, precisely distinguished the metastatic CRC samples from the non-metastatic ones. These genes have the potential to be used as biomarkers for the prognosis of metastatic CRC.
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Neoplasias Colorrectales/patología , Máquina de Vectores de Soporte , Línea Celular Tumoral , Neoplasias Colorrectales/genética , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Metástasis de la Neoplasia , Mapas de Interacción de Proteínas/genética , Control de Calidad , Reproducibilidad de los ResultadosRESUMEN
Many studies have demonstrated that the inflamed mucosa of Crohn's disease (CD) patients presented a disturbed gut commensal community, and the shift in microbial composition and species variety is associated with disease severity. To establish a link between changes in the intestinal bacterial composition and the alteration of inflammation, we obtained fecal bacteria from CD patients and non-CD controls. The bacteria were then used to stimulate the peripheral blood mononuclear cells (PBMCs) from one non-CD individual. We found that the frequency of IFN-γ- and IL-17-expressing CD4 T cells was significantly higher after stimulation with CD bacteria than with non-CD bacteria, while the frequency of IL-4- and IL-10-expressing CD4 T cells was significantly decreased after stimulation with CD bacteria. A similar trend was observed in the level of cytokine expression and transcription expression. However, this difference was not clear-cut, as overlapping regions were observed between the two groups. With longer stimulation using CD bacteria, the skewing toward Th1/Th17 responses were further increased. This increase depended on the presence of monocytes/macrophages. Interestingly, we also found that B cells presented an inhibitory effect in CD bacteria-mediated skewing toward Th1/Th17 cells and promoted IL-10 secretion in CD bacteria-stimulated PBMCs. Together, our results demonstrated that CD bacteria could promote Th1/Th17 inflammation in a host factor-independent fashion.
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Bacterias , Linfocitos T CD4-Positivos/inmunología , Enfermedad de Crohn/inmunología , Heces/microbiología , Células TH1/inmunología , Células Th17/inmunología , Adolescente , Adulto , Anciano , Linfocitos B/inmunología , Linfocitos B/microbiología , Linfocitos T CD4-Positivos/microbiología , Estudios de Casos y Controles , Células Cultivadas , Enfermedad de Crohn/microbiología , Citocinas/metabolismo , Femenino , Humanos , Leucocitos Mononucleares , Masculino , Persona de Mediana Edad , Células TH1/microbiología , Células Th17/microbiología , Adulto JovenRESUMEN
Colorectal cancer is the third most prevalent cancer type worldwide and contributes to a significant percentage of cancer-related mortality. Recent studies have shown that the CXCR5+CD8+ T cells present more potent proinflammatory function than CXCR5-CD8+ T cells in chronic virus infections and in follicular lymphoma, but the role of CXCR5+CD8+ T cells in colorectal cancer is yet unclear. In this study, we demonstrated that CXCR5+CD8+ T cells were very rare in peripheral blood mononuclear cells from healthy and colorectal cancer individuals, but were significantly enriched in resected tumors and tumor-associated lymph nodes. Compared to CXCR5-CD8+ T cells, the CXCR5+CD8+ T cells demonstrated significantly higher Bcl-6 expression and lower Blimp1 expression, suggesting that CXCR5+CD8+ T cells might represent a memory CD8+ T cell subset. CXCR5+CD8+ T cells also enhanced the IgG expression by autologous B cells. Under ex vivo condition, the CXCR5+CD8+ T cells demonstrated lower degranulation, TNFα expression and IFNγ expression than CXCR5-CD8+ T cells. However, after PMA + ionomycin stimulation, the degranulation and TNFα expression by CXCR5+CD8+ T cells were significantly elevated to a level comparable with CXCR5-CD8+ T cells, whereas the IFNγ expression by PMA + ionomycin-stimulated CXCR5+CD8+ T cells were significantly higher than that by CXCR5-CD8+ T cells. Following long-term TCR-stimulation, CXCR5+CD8+ T cells demonstrated significantly more potent proliferation capacity and higher IFNγ expression than CXCR5-CD8+ T cells. TCR-stimulated CXCR5+CD8+ T cells also showed a gradual downregulation in CXCR5 expression. We further found that TCR-stimulated CXCR5+CD8+ T cells demonstrated higher granzyme B production and induced more specific lysis of autologous tumor cells than CXCR5-CD8+ T cells. Together, these data demonstrate that CXCR5+CD8+ T cells represent a significant CD8+ T cell subset in colorectal tumors and have the potential to contribute to antitumor immunity, but their specific roles require further studies in vivo.
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Linfocitos T CD8-positivos/inmunología , Neoplasias Colorrectales/inmunología , Inmunoglobulina G/inmunología , Ganglios Linfáticos/inmunología , Receptores CXCR5/metabolismo , Subgrupos de Linfocitos T/inmunología , Anciano , Linfocitos B/inmunología , Proliferación Celular , Granzimas/biosíntesis , Humanos , Memoria Inmunológica/inmunología , Interferón gamma/biosíntesis , Ionomicina , Ganglios Linfáticos/citología , Persona de Mediana Edad , Factor 1 de Unión al Dominio 1 de Regulación Positiva , Proteínas Proto-Oncogénicas c-bcl-6/biosíntesis , Proteínas Represoras/biosíntesis , Acetato de Tetradecanoilforbol , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
BACKGROUND: Anterior resection syndrome (ARS) is common after sphincter-saving surgery for rectal cancer. It includes changes in the frequency and urgency of bowel movements and fecal incontinence. The therapeutic efficacy of biofeedback on ARS is unclear. We sought to evaluate the effectiveness of biofeedback therapy in patients with ARS after anterior resection for rectal cancer and to investigate the associated factors for therapeutic success. PATIENTS AND METHODS: The study was designed as a retrospective review of the data from 61 patients with ARS collected from a prospectively maintained institutional cancer database. Therapeutic efficacy was evaluated using anorectal manometry, the number of bowel movements daily, and fecal incontinence scoring systems (Vaizey and/or Wexner scores). Changes of > 15% in the Vaizey and/or Wexner scores were considered to indicate effectiveness. Stepwise logistic regression models were performed to evaluate whether the associated factors influenced therapeutic efficacy. RESULTS: The parameters of anorectal manometry in patients with rectal cancer were significantly lower than those in control group (P < .01). After biofeedback therapy, significant improvements were observed in the incontinence scale scores (P < .001), number of bowel movements (P < .001), and anorectal manometry data (maximum resting pressure, P < .001; maximum squeeze pressure, P = .001; and rectal capacity, P = .015). In contrast, no significant difference in the rectal initial sensation threshold was observed (P = .089). Patients with fecal incontinence as the primary symptom experienced significant improvements in all variables (P < .01), except for the rectal initial sensation threshold (P = .125). Age at surgery, current smoking status, diabetes, treatment cycles, laparoscopic surgery, interval from surgery to biofeedback therapy, and the use of radiation therapy were closely associated with therapeutic success. On multivariate analysis, current smoking status (odds ratio [OR], 0.09; 95% confidence interval [CI], 0.01-0.87), number of biofeedback therapy cycles (OR, 0.01; 95% CI, 0.00-0.06), and laparoscopic surgery (OR, 11.53; 95% CI, 1.17-113.61) were factors contributing to biofeedback therapeutic success. CONCLUSION: Biofeedback therapy can improve the anal function of patients after restorative resection for rectal cancer.
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Biorretroalimentación Psicológica , Procedimientos Quirúrgicos del Sistema Digestivo/efectos adversos , Enfermedades del Recto/etiología , Enfermedades del Recto/terapia , Neoplasias del Recto/cirugía , Adulto , Anciano , Femenino , Humanos , Masculino , Manometría , Persona de Mediana Edad , Estudios Retrospectivos , SíndromeRESUMEN
Resistance to chemotherapy is the major cause of colorectal cancer (CRC) treatment failure. The cytokine IL-22, which is produced by T cells and NK cells, is associated with tumorigenesis and tumor progression in cancers. However, the role of IL-22 in chemoresistance has not been investigated. We found that IL-22 levels in tumor tissues and peripheral blood were associated with chemoresistance and indicate poor prognosis for patients who received FOLFOX chemotherapy. In CRC cells, IL-22 was able to attenuate the cytotoxic and apoptosis-inducing effects of 5-FU and OXA by activating the STAT3 pathway and subsequently increasing the expression of anti-apoptotic genes. In addition, IL-22 conferred resistance to 5-FU and OXA by inducing IL-8 autocrine expression through STAT3 activation. Our findings identify IL-22 as a novel chemoresistance cytokine and may be a useful prognostic biomarker for CRC patients receiving FOLFOX chemotherapy.
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Antineoplásicos/uso terapéutico , Neoplasias Colorrectales/tratamiento farmacológico , Regulación Neoplásica de la Expresión Génica/fisiología , Interleucina-8/metabolismo , Interleucinas/metabolismo , Factor de Transcripción STAT3/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Comunicación Autocrina , Línea Celular Tumoral , Resistencia a Antineoplásicos , Fluorouracilo/uso terapéutico , Humanos , Interleucina-8/genética , Interleucinas/genética , Leucovorina/uso terapéutico , Compuestos Organoplatinos/uso terapéutico , Factor de Transcripción STAT3/genética , Insuficiencia del Tratamiento , Interleucina-22RESUMEN
MicroRNAs (miRNAs) are short, non-coding RNAs that regulate the expression of multiple target genes. Dysregulation of miRNAs is common in sepsis. Through microRNA microarray and qRT-PCR we found that the levels of miR-27a, miR-153 and miR-143 are up regulated, while let-7a, miR-218 and miR-129-5p are down regulated in lungs of septic mice. Knocking down of miR-27a down regulates expression levels of TNF-α and IL-6 significantly via reducing the phosphorylation level of NF-κB p65 and inhibiting its DNA binding activity. Furthermore, neutralisation of miR-27a up regulates PPARγ level, down regulates TNF-α expression, relieves pulmonary inflammation and promotes survival of septic mice, which demonstrates that miR-27a plays an important role in regulating inflammatory response in sepsis and provides a potential target for clinical sepsis research and treatment.
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Inflamación/genética , Inflamación/patología , MicroARNs/biosíntesis , Sepsis/genética , Sepsis/patología , Animales , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa , Sepsis/metabolismo , Regulación hacia ArribaRESUMEN
Mucin 1 (Muc1) is a tumor-associated glycoprotein and has an important role in cancer progression and metastasis. The aim of the study was to explore the effects and mechanism of Muc1 silencing on proliferation and tumor-forming capacity of colon cancer cell lines. In the present study, we first screened the colon cancer cell lines with high expression of Muc1 by western blot analysis. Then, the effective siRNA was selected and used for silencing endogenous Muc1. The results by MTT and vitro scratch assay showed that interference of Muc1 could effectively inhibit HCC2998 proliferation and migration (p<0.01). Also, colony-forming ability in Muc1-siRNA groups was significantly decreased compared with the control group (p<0.01). Cell cycle is detected by flow cytometry that suggested that Muc1-siRNA1 induced cell cycle arrest at S phase in HCC2998. Next, the expression and distribution of ß-catenin in cytoplasm and the nucleus was detected by western blot and the results revealed that the expression of ß-catenin was redistributed in Muc1-siRNA group. A higher ß-catenin level was detected in cytoplasm, while a lower ß-catenin level was located in nucleus, compared with controls (p<0.05). The tumorigenicity experiments showed that inhibition of Muc1 could significantly suppress the growth of HCC2998 in nude mice models (p<0.01). This study would effectively provide new clues for colon cancer therapy.
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Neoplasias del Colon/patología , Mucina-1/genética , Transducción de Señal , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Animales , Secuencia de Bases , Neoplasias del Colon/metabolismo , Ciclina D1/metabolismo , Cartilla de ADN , Progresión de la Enfermedad , Xenoinjertos , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The aim of the present study was to investigate whether the measurement of cytokines from abdominal exudate is valuable for the early diagnosis of prolonged postoperative ileus (PPOI) following colorectal surgery. In the present study, 100 consecutive patients who were scheduled to undergo elective resection for carcinoma of the sigmoid or rectum were investigated. Abdominal exudate was obtained via a drain tube following surgery for the detection of interleukin (IL)-1ß, IL-6 and tumour necrosis factor (TNF)-α. The correlation among the cytokine levels on postoperative days 1, 3 and 5 and the development of PPOI was investigated. Eight patients developed PPOI which was diagnosed 10-15 days postoperatively. No significant differences were observed among the peritoneal cytokine levels on postoperative days 1 and 3 in the 8 patients who developed PPOI when compared with those of the 92 patients that did not develop PPOI. By contrast, cytokine levels on postoperative day 5 were significantly higher in patients who developed PPOI when compared with those of patients that did not develop PPOI. The cytokine levels significantly increased during the first 5 days postoperatively in patients who developed PPOI and significantly decreased in patients who did not develop PPOI. The results of the present study showed that the increase in peritoneal IL-1ß, IL-6 and TNF-α levels may be an additional early diagnostic predictor of PPOI following colorectal surgery.
RESUMEN
BACKGROUND: Prolonged postoperative ileus (PPOI) is a common problem after major abdominal surgery and can cause postoperative morbidity, extended hospitalization, as well as increased health care costs. AIM: To investigate whether the levels of cytokines from abdominal exudate are predictive for early diagnosis of PPOI after colorectal surgery. METHODS: One hundred patients who had undergone elective resection for carcinoma of the sigmoid or rectum were recruited. The abdominal exudate was obtained from a drain tube after surgery to examine the levels of C-reactive protein (CRP), procalcitonin (PCT), and tumor necrosis factor alpha (TNF-α). The relationship between cytokine levels on postoperative day 1, 3 and 5 and the occurrence of PPOI was analyzed. RESULTS: Eight patients developed PPOI, which was diagnosed on postoperative day 10-15. PCT levels on postoperative day 1 and 3 were not significantly different between the 8 patients who developed PPOI and the 92 patients without PPOI. In contrast, PCT levels on day 5 were significantly higher in patients who developed PPOI than in patients without PPOI. The levels of CRP and TNF-α were irregular from day 1 to 5, and were not significantly different. CONCLUSIONS: Increased PCT levels in abdominal exudate may predict PPOI in the early period following colorectal surgery.
Asunto(s)
Calcitonina/metabolismo , Neoplasias Colorrectales/patología , Ileus/diagnóstico , Complicaciones Posoperatorias/diagnóstico , Precursores de Proteínas/metabolismo , Proteína C-Reactiva/metabolismo , Péptido Relacionado con Gen de Calcitonina , Neoplasias Colorrectales/complicaciones , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/cirugía , Diagnóstico Precoz , Exudados y Transudados/metabolismo , Humanos , Ileus/complicaciones , Ileus/metabolismo , Ileus/patología , Complicaciones Posoperatorias/patología , Periodo Posoperatorio , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
In this paper, an electrochemical acetylcholinesterase (AChE) inhibition assay based on three-dimensional ordered macroporous (3DOM) composite was conducted. The 3DOM composite was first fabricated on the glassy carbon electrode by electropolymerization of aniline in the presence of ionic liquid (IL) on a sacrificial silica nanospheres template. After the silica nanospheres were etched, an IL-doped polyaniline (IL-PANI) film with 3DOM morphology was formed. Then, gold nanoparticles (AuNPs) were decorated on the IL-PANI film by electrodeposition. The immobilized AChE on the 3DOM composite displayed favorable affinity to substrate acetylthiocholine chloride (ATCh), and the 3DOM composite showed excellent electrocatalytic effect on thiocholine, the hydrolysis product of ATCh. The presence of IL and AuNPs could improve the sensitivity by accelerating the electron transfer. The designed AChE biosensor was successfully applied to evaluate the AChE inhibition induced by endogenous neurotoxin 1(R),2N-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline [(R)-NMSal]. The results demonstrate that (R)-NMSal exerts a considerable effect on AChE activity, and the inhibition is reversible. The developed method offers a new approach for AChE inhibition assay, which is of great benefit in understanding the mechanism behind neurotoxin-induced neurodegenerative disorders.
Asunto(s)
Acetilcolinesterasa/metabolismo , Inhibidores de la Colinesterasa/farmacología , Electrophorus/metabolismo , Enzimas Inmovilizadas/metabolismo , Nanocompuestos/química , Tetrahidroisoquinolinas/farmacología , Compuestos de Anilina/química , Animales , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , PorosidadRESUMEN
In this paper, a novel dual enzymatic-biosensor is described for simultaneous determination of glucose and cholesterol in serum and peritoneal macrophages (PMs) of diabetic mice to evaluate the risk of diabetes-accelerated atherosclerosis. The biosensor was constructed by a three-step method. First, a poly-thionine (PTH) film was assembled on the surface of glassy carbon electrode by cyclic voltammetric electropolymerization of thionine, which serves as an electron transfer mediator (ETM). Second, gold nanoparticles (GNPs) were covered on the surface of PTH facilitating the electron transfer between glucose oxidase (GOx), cholesterol oxidase (ChOx) and electrode. Finally, the enzymes, GOx, cholesterol esterase (ChE), and ChOx, were covalently attached to the PTH layer through a chitosan (CH) linker. The PTH coupled with GNPs provides good selectivity, high sensitivity and little crosstalk for the dual enzymatic-biosensor. The developed biosensor had good electrocatalytic activity toward the oxidations of glucose and cholesterol, exhibiting a linear range from 0.008 mM to 6.0 mM for glucose with a detection limit of 2.0 µM, and a linear range from 0.002 mM to 1.0 mM for cholesterol with a detection limit of 0.6 µM. The results of the diabetic mice demonstrated that the cholesterol level did not change obviously with the increase of glucose level in serum, while the cholesterol level was induced with the increase of the glucose level in PMs. Previous studies have shown that the large accumulation of cholesterol in macrophage could lead to macrophage foam cell formation, which is the hallmark of early atherosclerosis. This study provides useful further evidences for the development of diabetes-accelerated atherosclerosis.