RESUMEN
BACKGROUND: We aimed to evaluate whether extracellular vesicles (EV)-derived microRNAs (miRNAs) can be used as biomarkers for advanced adenoma (AA) and colorectal cancer (CRC). METHODS: We detected the changes in the plasma EV-delivered miRNA profiles in healthy donor (HD), AA patient, and I-II stage CRC patient groups using miRNA deep sequencing assay. We performed the TaqMan miRNA assay using 173 plasma samples (two independent cohorts) from HDs, AA patients, and CRC patients to identify the candidate miRNA(s). The accuracy of candidate miRNA(s) in diagnosing AA and CRC was determined using the area under the receiver-operating characteristic curve (AUC) values. Logistic regression analysis was performed to evaluate the association of candidate miRNA(s) as an independent factor for the diagnosis of AA and CRC. The role of candidate miRNA(s) in the malignant progression of CRC was explored using functional assays. RESULTS: We screened and identified four prospective EV-delivered miRNAs, including miR-185-5p, which were significantly upregulated or downregulated in AA vs. HD and CRC vs. AA groups. In two independent cohorts, miR-185-5p was the best potential biomarker with the AUCs of 0.737 (Cohort I) and 0.720 (Cohort II) for AA vs. HD diagnosis, 0.887 (Cohort I) and 0.803 (Cohort II) for CRC vs. HD diagnosis, and 0.700 (Cohort I) and 0.631 (Cohort II) for CRC vs. AA diagnosis. Finally, we demonstrated that the upregulated expression of miR-185-5p promoted the malignant progression of CRC. CONCLUSION: EV-delivered miR-185-5p in the plasma of patients is a promising diagnostic biomarker for colorectal AA and CRC. Trial registration The study protocol was approved by the Ethics Committee of Changzheng Hospital, Naval Medical University, China (Ethics No. 2022SL005, Registration No. of China Clinical Trial Registration Center: ChiCTR220061592).
Asunto(s)
Adenoma , Neoplasias Colorrectales , Vesículas Extracelulares , MicroARNs , Humanos , Estudios Prospectivos , MicroARNs/genética , Adenoma/diagnóstico , Adenoma/genética , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/genéticaRESUMEN
Hepatocyte nuclear factor 4-α (HNF4α), a nuclear receptor, is expressed at lower levels in colon carcinoma tissues than in adjacent normal tissues. However, the relation between HNF4α and colon cancer progression and the underlying molecular mechanisms remain unclear. Here, we investigated the role of HNF4α in the progression of colon carcinoma. We showed that HNF4α mRNA and protein were downregulated in colon carcinoma specimens. HNF4α expression was related to pT classification (P < 0.001), lymph node metastasis (P = 0.002), distant metastasis (P < 0.001) and clinical stage (P < 0.001) in colon carcinoma patients. Patients with low or negative HNF4α expression had worse 3-year progression-free survival (PFS, P = 0.006) and overall survival (OS, P = 0.005) than patients with high HNF4α expression. Low HNF4α expression was an independent prognostic factor for 3-year PFS (hazard ratio 2.94; 95% confidence interval 1.047-8.250; P = 0.041). Ectopic expression of HNF4α inhibited colon carcinoma cell (HT29, LoVo, and SW480) proliferation, migration, and invasion, induced G2/M phase arrest and promoted apoptosis. Ectopic expression of HNF4α upregulated E-cadherin and downregulated vimentin in vitro, and suppressed SW480 xenograft tumor growth and liver metastasis in vivo. Furthermore, HNF4α overexpression downregulated the expression of snail, slug and twist. HNF4α inhibited EMT through its effect on the Wnt/ß-catenin signaling pathway, and HNF4α downregulation may be mediated by promoter methylation in cancer tissues. Our results suggest that downregulation of HNF4α plays a critical role in the aggravation of colon carcinoma possibly by promoting EMT via the Wnt/ß-catenin signaling pathway and by affecting apoptosis and cell cycle progression.
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Neoplasias del Colon/genética , Neoplasias del Colon/patología , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/metabolismo , Animales , Apoptosis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Neoplasias del Colon/metabolismo , Progresión de la Enfermedad , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , Femenino , Regulación Neoplásica de la Expresión Génica , Células HT29 , Humanos , Masculino , Ratones , Metástasis de la Neoplasia , Trasplante de Neoplasias , Análisis de Supervivencia , beta Catenina/metabolismoRESUMEN
BACKGROUND: The aim of the present study was to investigate the therapeutic effects of p75 tumor necrosis factor receptor monoclonal antibody D8F2 on a traumatic arthritis model in rats, and to explore the underlying mechanism. METHODS: Forty male Sprague Dawley rats were randomly divided into five groups: (A) sham operation control group, (B) traumatic arthritis model group, (C) low-dose D8F2 group (1 mg/kg), (D) medium-dose D8F2 group (3 mg/kg), and (E) high-dose D8F2 group (10 mg/kg). Joint fluid samples were collected at 72 h after surgery, and enzyme-linked immunosorbent assay was performed to measure the following inflammatory factors: tumor necrosis factor α (TNF-α) and interleukin 1ß. One week after the surgery, rats were killed, and immunohistochemical staining was applied to detect the matrix metalloproteinase (MMP1 and MMP3) expression in the synovium. In cultured synovial fibroblast experiments, the D8F2-induced ubiquitination of TNF receptor-associated factor 2 (TRAF2) was examined by immunoprecipitation, and nuclear translocation of p65 nuclear factor-κB (p65NF-κB) mediated by TNF-α and D8F2 was analyzed by western blotting. RESULTS: In the traumatic arthritis model group, the inflammatory factors and MMPs were significantly increased relative to the sham operation control group (P < 0.05), whereas D8F2 could downregulate these factors in a dose-dependent manner (P < 0.05). The results from in vitro studies indicated that D8F2 can induce TRAF2 ubiquitination and inhibit the nuclear translocation of p65NF-κB mediated by TNF-α. CONCLUSIONS: p75 Tumor necrosis factor receptor monoclonal antibody has a therapeutic effect on traumatic arthritis, which may occur via the downregulation of inflammatory factors and MMPs at the transcription level because of TRAF2 degradation and inhibited activation of NF-κB.
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Anticuerpos Monoclonales/uso terapéutico , Artritis/tratamiento farmacológico , Articulaciones/lesiones , Receptores Tipo II del Factor de Necrosis Tumoral/antagonistas & inhibidores , Animales , Citocinas/análisis , Modelos Animales de Enfermedad , Masculino , Metaloproteinasa 1 de la Matriz/análisis , FN-kappa B/antagonistas & inhibidores , Ratas , Ratas Sprague-Dawley , Receptores Tipo II del Factor de Necrosis Tumoral/fisiología , Factor 2 Asociado a Receptor de TNF/metabolismoRESUMEN
OBJECTIVE: To investigate the effects of and mechanisms underlying the activation of the p75 tumor necrosis factor receptor (p75TNFR) signaling pathway in inflammatory responses in mice with traumatic brain injury. METHODS: We first generated hybridomas that produced antibodies specific for p75TNFR, by inoculating BALB/c mice with antigenic peptides derived from mouse p75TNFR, which is critical to the binding of tumor necrosis factor-alpha (TNF-α) and p75TNFR. The isotype, epitope, titer, specificity, and affinity constant of monoclonal antibodies (mAbs) were determined using commercial kits and enzyme-linked immunosorbent assay. We then screened the agonist antibody via L929 cytotoxicity assay. The levels of inflammatory factors were detected in C57BL/6 mice with traumatic brain injury and then the mice were injected with either saline (control) or p75TNFR agonist mAb. Furthermore, we investigated the effects of p75TNFR agonist mAb on p38MAPK and nuclear factor-κB signals. RESULTS: Seven mAbs against p75TNFR were generated. Among them, the mAb D8F2 could markedly enhance the cytotoxicity of TNF-α on L929 cells. In a traumatic brain injury model, D8F2 could inhibit the levels of inflammatory factors and downregulate RNA transcription of these factors by suppressing the activation of p38 mitogen-activated protein kinase and nuclear factor-κB. CONCLUSION: The mAb D8F2 could inhibit posttraumatic inflammatory responses effectively. In this study, we developed an agonist anti-mouse p75TNFR mAb, which may be used in the future to devise new strategies for the clinical treatment of inflammation after trauma.
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Anticuerpos Monoclonales/uso terapéutico , Lesiones Encefálicas/complicaciones , Encefalitis/etiología , Encefalitis/prevención & control , Receptores Tipo II del Factor de Necrosis Tumoral/antagonistas & inhibidores , Transducción de Señal/fisiología , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Lesiones Encefálicas/metabolismo , Modelos Animales de Enfermedad , Encefalitis/metabolismo , Femenino , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Receptores Tipo II del Factor de Necrosis Tumoral/inmunología , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
BACKGROUND: The cause of ulcerative colitis (UC) is not yet fully understood. Previous research has pointed towards a potential role for mutations in nucleotide-binding oligomerization domain-containing protein 2 (NOD2) in promoting the onset and progression of inflammatory bowel disease (IBD) by altering the microbiota of the gut. However, the relationship between toll-like receptor 4 (TLR4) and gut microbiota in IBD is not well understood. To shed light on this, the interaction between TLR4 and gut microbiota was studied using a mouse model of IBD. METHODS: To examine the function of TLR4 signaling in intestinal injury repair, researchers developed Dextran Sulfate Sodium Salt (DSS)-induced colitis and injury models in both wild-type (WT) mice and TLR4 knockout (TLR4-KO) mice. To assess changes in the gut microbiota, 16S rRNA sequencing was conducted on fecal samples from both the TLR4-KO and WT enteritis mouse models. RESULTS: The data obtained depicted a protective function of TLR4 against DSS-induced colitis. The gut microbiota composition was found to vary considerably between the WT and TLR4-KO mice groups as indicated by ß-diversity analysis and operational taxonomic units (OTUs) cluster. Statistical analysis of microbial multivariate variables depicted an elevated abundance of Escherichia coli/Shigella, Gammaproteobacteria, Tenerlcutes, Deferribacteres, Enterobacteria, Rikenellaceae, and Proteobacteria in the gut microbiota of TLR4-KO mice, whereas there was a considerable reduction in Bacteroidetes at five different levels of the phylogenetic hierarchy including phylum, class, order, family, and genus in comparison with the WT control. CONCLUSIONS: TLR4 may protect intestinal epithelial cells from damage in response to DSS-induced injury by controlling the microbiota in the gut.
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Colitis , Microbioma Gastrointestinal , Enfermedades Inflamatorias del Intestino , Receptor Toll-Like 4 , Animales , Ratones , Colitis/inducido químicamente , Colitis/genética , Células Epiteliales , Escherichia coli , Filogenia , ARN Ribosómico 16S/genética , Cloruro de Sodio Dietético , Receptor Toll-Like 4/genéticaRESUMEN
BACKGROUND: Venous thromboembolism (VTE) is a common and serious complication after colorectal cancer (CRC) surgery. Few large-sample studies have reported VTE incidence and management status after CRC surgery in China. This study aimed to investigate the incidence and prevention of VTE in Chinese patients after CRC surgery, identify risk factors for developing VTE, and construct a new scoring system for clinical decision-making and care planning. METHODS: Participants were recruited from 46 centers in 17 provinces in China. Patients were followed up for 1 month postoperatively. The study period was from May 2021 to May 2022. The Caprini score risk stratification and VTE prevention and incidence were recorded. The predictors of the occurrence of VTE after surgery were identified by multivariate logistic regression analysis, and a prediction model (CRC-VTE score) was developed. RESULTS: A total of 1836 patients were analyzed. The postoperative Caprini scores ranged from 1 to 16 points, with a median of 6 points. Of these, 10.1% were classified as low risk (0-2 points), 7.4% as moderate risk (3-4 points), and 82.5% as high risk (≥5 points). Among these patients, 1210 (65.9%) received pharmacological prophylaxis, and 1061 (57.8%) received mechanical prophylaxis. The incidence of short-term VTE events after CRC surgery was 11.2% (95% CI 9.8-12.7), including deep venous thrombosis (DVT) (11.0%, 95% CI 9.6-12.5) and pulmonary embolism (PE) (0.2%, 95% CI 0-0.5). Multifactorial analysis showed that age (≥70 years), history of varicose veins in the lower extremities, cardiac insufficiency, female sex, preoperative bowel obstruction, preoperative bloody/tarry stool, and anesthesia time at least 180 min were independent risk factors for postoperative VTE. The CRC-VTE model was developed from these seven factors and had good VTE predictive performance ( C -statistic 0.72, 95% CI 0.68-0.76). CONCLUSIONS: This study provided a national perspective on the incidence and prevention of VTE after CRC surgery in China. The study offers guidance for VTE prevention in patients after CRC surgery. A practical CRC-VTE risk predictive model was proposed.
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Neoplasias Colorrectales , Embolia Pulmonar , Tromboembolia Venosa , Humanos , Femenino , Anciano , Tromboembolia Venosa/epidemiología , Tromboembolia Venosa/etiología , Tromboembolia Venosa/prevención & control , Estudios Prospectivos , Incidencia , Pueblos del Este de Asia , Medición de Riesgo , Factores de Riesgo , Embolia Pulmonar/complicaciones , Neoplasias Colorrectales/cirugía , Neoplasias Colorrectales/complicaciones , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/prevención & controlRESUMEN
BACKGROUND: The objectives of this study were to identify and validate the diagnostic value of N-glycan markers in colorectal cancer (CRC) and to uncover their underlying molecular mechanism. METHODS: In total, 347 individuals, including patients with CRC, patients with colorectal adenoma, and healthy controls, were divided randomly into a training group (n = 287) and retrospective validation groups (n = 60). Serum N-glycan profiling was analyzed by DNA sequencer-assisted/flurophore-assisted carbohydrate electrophoresis (DSA-FACE). Two diagnostic models were constructed based on N-glycan profiling with logistic stepwise regression. The diagnostic performance of each model was assessed further in retrospective, prospective (n = 43), and follow-up (n = 46) cohorts. Lectin blot and reverse transcriptase-polymerase chain reaction were used to analyze the total core-fucosylated residues and molecular expression involved in core-fucosylation modifications in CRC. RESULTS: Two diagnostic models designated CRCglycoA and CRCglycoB were constructed to differentiate CRC from normal and adenoma, respectively. The areas under the receiver operating characteristic curves (AUC) of both CRCglycoA and CRCglycoB were higher than the AUC of carcinoembryonic antigen (CEA) (CRCglycoA, 0.92 vs 0.81; CRCglycoB, 0.81 vs 0.73). The sensitivity and accuracy of CRCglycoA improved from 21.7% to 25% and from 11.63% to 18% in the training cohort, the retrospective cohort, and the prospective cohorts compared with the sensitivity and accuracy of CEA. The sensitivity of CRCglycoB improved from 20% to 28.23%. Both altered N-glycans, and results from the diagnostic models were reversed after curative surgery. The level of total core fucose residues and fucosyltransferase were decreased significantly in CRC. CONCLUSIONS: The current results indicated that the N-glycan markers based diagnostic models are new, valuable, noninvasive alternatives for identifying CRC. The authors concluded that decreased fucosyltransferase may be responsible for decreased levels of total core-fucosylated modification in both tissues and serum from patients with CRC.
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Adenoma/diagnóstico , Biomarcadores de Tumor/sangre , Neoplasias Colorrectales/diagnóstico , Lectinas/sangre , Polisacáridos/sangre , Adenoma/sangre , Estudios de Casos y Controles , Neoplasias Colorrectales/sangre , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: We hypothesize that introduction of nano-silver particles to porcine-derived small intestinal submucosa (NS-PSIS) would lead to significant enhancement in antibacterial property in repairing contaminated abdominal defect. BACKGROUND: Porcine-derived small intestinal submucosa (PSIS) is an acellular and xenogenic biological material intensively used in repairing and regenerating wounded and dysfunctional tissues. Surgical site infection (SSI) remains so far a serious problem and major challenge, particularly in contaminated tissue-deficient repairing. METHODS: Self-assembly was used to fabricate NS-PSIS. The antibacterial property was evaluated in vitro and in vivo by means of repairing full-thickness contaminated abdominal defect in rats. The native PSIS and polypropylene-oxidized regenerated cellulose were served as controls. In addition, changes in biomechanical resistance, morphology and immunohistochemistry for inflammatory reaction and neovasculation in the repaired abdominal wall were analyzed. Biosafety was investigated by pyrogen test, skin irritation test and silver measurement in vivo. RESULTS: NS-PSIS exhibited strong antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa on agar diffusion, with mean diameters of inhibition zone ranging from 11.9 to 23.5 mm. There were significantly lower SSI incidence and a tendency of better abdominal wall resistance in the NS-PSIS group as compared with the PSIS or polypropylene-oxidized regenerated cellulose group after repairing contaminated abdominal defect in rats. Nano-silver modified PSIS did not change the native PSIS property in the tissue recolonization, remodeling and neovascularization. NS-PSIS was not pyrogenic or skin irritated, without silver residual in vivo after repairing contaminated abdominal defect. CONCLUSION: Nano-silver particles to PSIS lead to significant enhancements in antibacterial property in vitro and in vivo without decreasing its biomechanical resistance and biocompatibility. This study provides proof of concept for the use of nano-silver modified naturally derived PSIS as an ideal scaffold for SSI prevention in the contaminated tissue-deficient repair.
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Antibacterianos/farmacología , Mucosa Intestinal/trasplante , Plata/farmacología , Colgajos Quirúrgicos , Infección de la Herida Quirúrgica/cirugía , Pared Abdominal/cirugía , Animales , Apósitos Biológicos , Modelos Animales de Enfermedad , Yeyuno/cirugía , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Medición de Riesgo , Infección de la Herida Quirúrgica/microbiología , Infección de la Herida Quirúrgica/prevención & control , Porcinos , Resistencia a la Tracción , Ingeniería de Tejidos , Cicatrización de Heridas/fisiologíaRESUMEN
Bim is a proapoptotic member of the Bcl-2 family and is primarily involved in the regulation of the intrinsic apoptotic pathway. However, the detail of regulation of Bim's proapoptotic activity has not been clarified yet. Using Bim L as bait, we screened a human fetal cDNA library for interacting proteins and identified Grb10 as an interactor. This interaction was verified by co-immunoprecipitation and intracellular co-localization studies. The potential segment of Bim L that binds Grb10 was identified via a yeast mating test. Grb10 interacted with the DBD (dynein binding domain) of Bim and inhibited apoptosis triggered by overexpression of DBD containing Bim isoforms. The putative phosphorylation sites on DBD of Bim play a role for the anti-proapoptotic activity of Grb10. Our results suggest that Grb10 interacts with Bim L and inhibits its proapoptotic activity in a phosphorylation-dependant manner.
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Proteínas Reguladoras de la Apoptosis/metabolismo , Apoptosis , Proteína Adaptadora GRB10/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Proteína 11 Similar a Bcl2 , Células HEK293 , Humanos , Inmunoprecipitación , Espacio Intracelular/metabolismo , Unión Proteica , Isoformas de Proteínas/metabolismo , Transporte de Proteínas , Proteínas Recombinantes de Fusión/metabolismo , Saccharomyces cerevisiae/metabolismo , Fracciones Subcelulares/metabolismo , Técnicas del Sistema de Dos HíbridosRESUMEN
Recently, homologous pleckstrin-homology (PH)-domain leucine-rich-repeat protein phosphatases (PHLPP2) has been reported as a tumor suppressor in colon cancer. This study aimed to unravel the possible involvement of long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) regulating PHLPP2 in colon cancer. Expressions of candidate lncRNAs and miRNAs were verified by the RT-qPCR and Western blot analyses in colon cancer. The roles of candidate genes in colon cancer were investigated in HT-29 cells in vitro and in mouse tumor xenograft model in vivo. PHLPP2, a target of miR-141 and miR-424, was downregulated in colon cancer. PHLPP2 upregulation and miR-141 and miR-424 downregulation suppressed the colon cancer cell proliferation, migration, invasion, and epithelial-mesenchymal transition, and promote cell apoptosis, which also resulted in suppression of tumor metastasis and formation. Furthermore, LINC00402, LINC00461, and SFTA1P were identified as the targets of miR-141 and miR-424 and acted as competitive endogenous RNAs (ceRNAs) of PHLPP2. The upregulation of LINC00402, LINC00461, and SFTA1P was verified to enhance the suppressive effects of PHLPP2 in the pathogenesis of colon cancer. Conjointly, our results demonstrated the suppressive effects of PHLPP2 in colon cancer and proved that LINC00402, LINC00461, and SFTA1P acted as ceRNAs of PHLPP2 by competitive binding to miR-141 and miR-424.
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Neoplasias del Colon , MicroARNs/metabolismo , Fosfoproteínas Fosfatasas/metabolismo , ARN Largo no Codificante/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Animales , Proliferación Celular , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Femenino , Células HT29 , Xenoinjertos , Humanos , Masculino , Ratones , Ratones Desnudos , Persona de Mediana Edad , Metástasis de la Neoplasia/patología , Metástasis de la Neoplasia/fisiopatologíaRESUMEN
PURPOSE: To evaluate the efficiency, safety, and outcome of laparoscopic adhesiolysis for recurrent small-bowel obstruction (SBO), when performed early after failed conservative treatment. METHODS: Between 1999 and 2005, elective laparoscopic adhesiolysis was attempted in 46 patients with recurrent SBO after abdominal or pelvic surgery. Laparoscopic adhesiolysis was done during the acute onset of SBO after the patient failed to respond to 24 h of conservative treatment. RESULTS: Fifteen patients (32.6%) presented with recurrent SBO and 31 patients (67.4%) presented with recurrent SBO and chronic abdominal pain. Postoperative adhesions were identified laparoscopically in all patients: as isolated bands in 11 patients, enteroperitoneal angulation in 12 patients, entero-enteral angulation in 17 patients, and extensive dense and matted intra-abdominal adhesions in 6 patients. Successful complete laparoscopic adhesiolysis was achieved in 42 of the 46 patients (91.3%). Conversion to minilaparotomy was required for a convoluted mass of adherent bowel in one patient (2.2%) and laparotomy was required for extensive dense and matted adhesions in three patients (6.5%). The mean follow-up was 46.5 months (range 24-89 months). Forty-three patients (93.5%) were asymptomatic after the operation. Only one patient (2.2%) had a further two episodes of SBO over 38 months of follow-up. CONCLUSION: Laparoscopic intervention, when done early after the onset of symptoms, is highly feasible, safe, and effective in selected patients with recurrent SBO caused by postoperative adhesion.
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Obstrucción Intestinal/etiología , Obstrucción Intestinal/cirugía , Intestino Delgado/cirugía , Laparoscopía , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/cirugía , Adherencias Tisulares/complicaciones , Adherencias Tisulares/cirugía , Abdomen Agudo/etiología , Abdomen Agudo/cirugía , Adulto , Anciano , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Recurrencia , Resultado del TratamientoRESUMEN
OBJECTIVE: To investigate the effects of Baihe Recipe, a compound traditional Chinese herbal medicine, on growth and metastasis of orthotopically transplanted gastric carcinoma and the expressions of vascular endothelial growth factor (VEGF) and p53 proteins in the tumor tissues in nude mice. METHODS: Forty-five nude mice orthotopically transplanted with BGC-823 human gastric cancer cells were randomly divided into three groups: Baihe Recipe group, 5-fluorouracil (5-FU) group and untreated group. The mice in the Baihe Recipe group received intragastric administration of 0.5 mL Baihe Recipe (crude drug content was 0.2 g/mL) for 6 weeks, and the mice in the untreated group received 0.5 mL normal saline. The mice in the 5-FU group received an intraperitoneal injection of 5-FU dilution (0.2 mL, 60 mg/kg per week, for 3 weeks). All mice were sacrificed after 6-week treatment. The weights of tumor and the growth-inhibiting rate were measured and the expressions of VEGF and p53 proteins were detected by immunohistochemical method. RESULTS: The growth inhibition rates in the Baihe Recipe and 5-FU groups were 52.86% and 42.87% respectively. The incidence rates of metastasis to perigastric and hepatic portal lymph nodes, and to liver and peritoneum in the Baihe Recipe and 5-FU groups were lower than those in the untreated group. The metastasis rates in Baihe Recipe group, 5-FU group and untreated group were 33.33%, 35.71% and 80.00% respectively, with significant difference (P<0.05), and the expressions of VEGF and p53 proteins in tumor tissues in the Baihe Recipe group were lower than those in the untreated group and the 5-FU group (P<0.01, P<0.05). CONCLUSION: Baihe Recipe has inhibitory effects on tumor growth and metastasis of gastric cancer orthotopically transplanted in nude mice by down-regulating the expressions of VEGF and p53 proteins.
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Medicamentos Herbarios Chinos/uso terapéutico , Fitoterapia , Neoplasias Gástricas/tratamiento farmacológico , Proteína p53 Supresora de Tumor/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Animales , Antineoplásicos Fitogénicos/uso terapéutico , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Distribución Aleatoria , Proteína p53 Supresora de Tumor/genética , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
BACKGROUND: The metastasis of colorectal cancer frequently tends to liver, which is one of the three leading causes of cancer-related deaths worldwide. Growing evidence showed that aspirin could effectively inhibit liver metastasis of colorectal cancer. However, the potential mechanism has not been fully understood. METHODS: Mouse splenic vein metastasis assay was used to examine the metastatic ability of colon cancer cells in vivo. And wound healing and transwell assay were applied to detect the metastasis potential of C26 and HCT116 colon cancer cell lines in vitro. RT-PCR and western blotting were used to explore Toll-like receptor 4 (TLR4) expression in colon cancer cell lines. The functions of TLR4 in the migration of the colon cancer cell line were analyzed by infecting cells with lentivirus containing TLR4 siRNA. RESULTS: We demonstrated that lipopolysaccharides (LPS) could enhance the metastasis potential of C26 and HCT116 colon cancer cell lines. However, aspirin effectively decreased the metastasis capacity of colon cancer cells in vitro and in vivo. We found that the enhancement of LPS on the migration of colon cancer cells by inducing epithelial-mesenchymal transition (EMT) phenotype demonstrated a TLR4-dependent manner. Aspirin treatment lead to the downregulation of TLR4 on C26 cells which resulted in the decrease of C26 cells migration and EMT phenotype that induced by LPS. Additionally, the inhibitory effect from aspirin on the expression of TLR4 on C26 cells leads to the downregulation of NF-κB. CONCLUSION: The results of our study indicate that LPS origin from intestinal flora may promote the metastasis of colon cancer to liver and aspirin may inhibit the metastasis of colon cancer by inhibiting the expression of TLR4.
RESUMEN
B-cell CLL/lymphoma 9 protein (BCL-9), a multi-functional co-factor in Wnt signaling, induced carcinogenesis as well as promoting tumor progression, metastasis and chemo-resistance in colorectal cancer (CRC). However, the mechanisms for increased BCL-9 expression in CRC were not well understood. Here, we report that hypoxia, a hallmark of solid tumors, induced BCL-9 mRNA expression in human CRC cells. Analysis of BCL-9 promoter revealed two functional hypoxia-responsive elements (HRE-B and HRE-C) that can be specifically bound with and be transactivated by hypoxia inducible factors (HIF) -1α but not HIF-2α. Consistently, ectopic expression of HIF-1α but not HIF-2α transcriptionally induced BCL-9 expression levels in cells. Knockdown of endogenous HIF-1α but not HIF-2α by siRNA largely abolished the induction of HIF by hypoxia. Furthermore, there was a strong association of HIF-1α expression with BCL-9 expression in human CRC specimens. In summary, results from this study demonstrated that hypoxia induced BCL-9 expression in human CRC cells mainly through HIF-1α, which could be an important underlying mechanism for increased BCL-9 expression in CRC.
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Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Hipoxia/genética , Hipoxia/metabolismo , Proteínas de Neoplasias/genética , Anciano , Anciano de 80 o más Años , Línea Celular Tumoral , Neoplasias Colorrectales/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia , Estadificación de Neoplasias , Regiones Promotoras Genéticas , Elementos de Respuesta , Factores de Transcripción , Activación TranscripcionalRESUMEN
There exists a great need for repair grafts with similar volume to human skeletal muscle that can promote the innate ability of muscle to regenerate following volumetric muscle loss. Perfusion decellularization is an attractive technique for extracellular matrix (ECM) scaffold from intact mammalian organ or tissue which has been successfully used in tissue reconstruction. The perfusion-decellularization of skeletal muscle has been poorly assessed and characterized, but the bioactivity and functional capacity of the obtained perfusion skeletal muscle ECM (pM-ECM) to remodel in vivo is unknown. In the present study, pM-ECM was prepared from porcine rectus abdominis (RA). Perfusion-decellularization of porcine RA effectively removed cellular and nuclear material while retaining the intricate three-dimensional microarchitecture and vasculature networks of the native RA, and many of the bioactive ECM components and mechanical properties. In vivo, partial-thickness abdominal wall defects in rats repaired with pM-ECM showed improved neovascularization, myogenesis and functional recellularization compared to porcine-derived small intestinal submucosa (SIS). These findings show the biologic potential of RA pM-ECM as a scaffold for supporting site appropriate, tissue reconstruction, and provide a better understanding of the importance maintaining the tissue-specific complex three-dimensional architecture of ECM during decellularization and regeneration.
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Traumatismos Abdominales/terapia , Bioprótesis , Matriz Extracelular/trasplante , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/trasplante , Andamios del Tejido/química , Abdomen/irrigación sanguínea , Abdomen/patología , Traumatismos Abdominales/patología , Traumatismos Abdominales/cirugía , Animales , Fenómenos Biomecánicos , Línea Celular , Matriz Extracelular/química , Femenino , Músculo Esquelético/química , Perfusión , Ratas Sprague-Dawley , Porcinos , Ingeniería de TejidosRESUMEN
In this dataset, we particularly depicted the harvest and perfusion decellularization of porcine rectus abdominis (RA), accompanied with displaying of the retained vascular trees within the perfusion-decellularized skeletal muscle matrix (pM-ECM) using vascular corrosion casting. In addition, several important tips for successful pM-ECM preparation were emphasized, which including using anatomically isolated skeletal muscle as tissue source with all main feeding and draining vessels perfused, preserving the internal microcirculation availability, aseptic technique and pyrogen free in all steps, sequential perfusion via artery or vein, and longtime washing after decellularization. The data are supplemental to our original research article describing detailed associations of pM-ECM as a clinically relevant scale, three-dimensional scaffold with a vascular network template for tissue-specific regeneration, "Perfusion-decellularized skeletal muscle as a three-dimensional scaffold with a vascular network template" Zhang et al. (2016) [1].
RESUMEN
Gallbladder cancer (GBC) is the most common malignant tumor of the biliary system. However, the mechanisms underlying its tumor initiation, progression, and metastasis are not yet fully understood. The annexin A4 (ANXA4) gene is highly expressed in GBC tissues and may play an important role in the initiation and progression of this disease. In this study, we examined the up-regulation of ANXA4 in human GBC tissues and cell lines. Elevated ANXA4 correlated well with invasion depth in GBC patients and predicted a poor prognosis. In vitro, GBC-SD and NOZ cells with ANXA4 knockdown demonstrated increased apoptosis and inhibited cell growth, migration, and invasion. Interactions between ANXA4 and nuclear factor-κB (NF-κB) p65 proteins were detected. In vivo, ANXA4 knockdown inhibited tumor growth of GBC cells in nude mice and down-regulated the expression of downstream factors in the NF-κB signaling pathway. Taken together, these data indicate that up-regulation of ANXA4 leads to activation of the NF-κB pathway and its target genes in a feedback regulatory mechanism via the p65 subunit, resulting in tumor growth in GBC.
Asunto(s)
Anexina A4/metabolismo , Neoplasias de la Vesícula Biliar/patología , Factor de Transcripción ReIA/metabolismo , Animales , Línea Celular Tumoral , Regulación de la Expresión Génica , Humanos , Ratones Desnudos , Unión ProteicaRESUMEN
OBJECTIVE: To establish a non-invasive model for the assessment of portal venous pressure (PVP) based on the magnetic resonance (MR) parameters. METHODS: In this prospective study, contrast-enhanced magnetic resonance imaging (MRI) scan was performed in 109 patients indicated for upper abdominal surgeries after their written consents were obtained, and intraoperative PVP measurements were completed in 92 patients. Altogether 17 patients were excluded for not undergoing surgery or unsuccessful catheterization. A linear model was constructed for estimating PVP levels in 56 patients and further validation was conducted in the other 36 patients. RESULTS: The PVP levels were significantly correlated with MR parameters, including splenic volume (SV), splenic venous diameter (SVD), liver/splenic volume ratio, portal venous diameter, hepatic diameter, portal venous cross-sectional area, ascites, varices and arterial portal shunts. A linear model was established as follows: PVP (mmHg) = 2.529 + 1.572 × SVD (mm) + 0.231 × SV/body mass index (× 10(4) cm(5) /kg) + 3.44 × aspartate aminotransferase-to-platelet ratio index. This model showed excellent accuracy in the detection of portal hypertension, with the area under the receiver operating characteristic curve (AUROC) of 0.945 (95% CI 0.867-1.000), with the sensitivity and specificity of 91.7% and 93.7%, respectively. The agreement analysis revealed that the predictive value using this formula closely reflected the patients' actual PVP level. Moreover, the validation confirmed the accuracy of this model for the assessment of portal hypertension [AUROC 0.935 (95% CI 0.856-1.000)]. CONCLUSIONS: The MRI-based formula has great potential for detecting portal hypertension. As a non-invasive measurement, it may be clinically accepted for the replacement of invasive modalities after further refinement.
Asunto(s)
Hipertensión Portal/diagnóstico por imagen , Modelos Cardiovasculares , Vena Porta/diagnóstico por imagen , Adulto , Anciano , Medios de Contraste , Femenino , Humanos , Hipertensión Portal/patología , Hipertensión Portal/fisiopatología , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Variaciones Dependientes del Observador , Presión Portal , Vena Porta/patología , Vena Porta/fisiopatología , Valor Predictivo de las Pruebas , Estudios Prospectivos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Bazo/patología , Vena Esplénica/diagnóstico por imagen , Vena Esplénica/patologíaRESUMEN
BACKGROUND: Perioperative allogeneic blood transfusion (ABT) may be a deleterious predictor on the prognosis of gastric cancer (GC) for subjects who had undergone curative surgeries. In this article we proposed to figure out the effect of ABT with a systematic review and meta-analysis. METHODS: Relevant articles were identified by searching Pubmed and Embase to March 2014. A random-effects model or fixed-effects model was used to calculate pooled odds ratios (ORs). Sensitivity analysis, meta-regression, stratified analysis, dose-response meta-analysis were conducted, and publication bias tested. RESULTS: Eighteen studies (9120 GC patients) were included, of which 36.3% received transfusions. ABT was associated with increased all-cause mortality (OR, 2.17; 95% confidence interval [CI], 1.72-2.74; p<0.001; I2=75%). Sensitivity analysis showed significant changes in ORs while meta-regression had little influence on ORs. Galbraith plot revealed the OR reduced to 2.10 (95% CI, 1.86-2.37; p<0.001) with tau2 reduced to 0.00 and I2 reduced to 0%. RESULTS of stratified analysis were robust and consistent. Dose-response meta-analysis revealed that all-cause mortality was significantly lower in patients transfused with ≤800 mL of blood than those transfused with >800 mL (OR, 0.58; 95% CI, 0.37-0.92; p=0.02; I2=54%). ABT was also associated with increased cancer-related mortality (OR, 2.57, p=0.011) and recurrence (OR, 1.52, p=0.017). CONCLUSIONS: In GC patients undergoing curative surgeries, ABTs are associated with a worse prognosis, including all-cause mortality, cancer-related mortality and recurrence. Patient blood management should be investigated further to minimize use of ABT.
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Transfusión Sanguínea/mortalidad , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/cirugía , Reacción a la Transfusión , Causas de Muerte , Humanos , Recurrencia Local de Neoplasia , Oportunidad Relativa , Pronóstico , Factores de Riesgo , Resultado del TratamientoRESUMEN
OBJECTIVE: Metachronous bone metastasis (MBM) occurs in 6-10% of colorectal cancer (CRC) patients following surgical treatment. The aim of this study is to determine the risk factors affecting the development of MBM in CRC patients following curative resection. METHOD: Clinical and pathological records of 516 CRC patients who underwent curative resection were retrospectively studied. The association between clinicopathological variables and development of MBM was investigated using univariate and multivariate analyses. RESULT: The incidence of MBM was 6.0% and the median time of developing MBM was 15 (range, 1-89) months. Univariate analysis identified that lymph node involvement (p = 0.001), tumor stage (p = 0.020) and tumor location (p = 0.015) were significantly correlated with development of MBM. Multivariate analyses showed tumor location (p = 0.039) and lymph node involvement (p = 0.003) were independent risk factors contributing to the occurrence of MBM. CONCLUSION: This study indicated that tumor location and lymph node involvement were independent risk factors for development of MBM in CRC patients after curative resection.