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Mesenchymal stem cells (MSCs) derived exosomes (Exos) are one of the most promising candidate for the treatment of this condition. However, the underlying molecular mechanism remains uncertain. Here we investigated the therapeutic effect of exosomal miR-181c-5p (ExomiR-181c-5p) on a rat model of neuropathic pain induced by sciatic nerve chronic constriction injury (CCI). In this study NP model was established using the CCI method. NP levels were assessed using PWT and PWL. Microarray analysis and RT-PCR were used to determine the relative expression of miR-181c-5p. MSC-derived exosomes were extracted using the total exosome isolation reagent characterized by WB and NTA. MiR-181c-5p was loading into Exos using electroporation. The inflammation response in microglia cells and CCI rats were assessed by ELISA assay respectively. Our study demonstrates that miR-181c-5p expression was obviously decreased in a time-dependent manner in CCI rats. MiR-181c-5p was effectively electroporated and highly detected in MSC-derived Exos. ExomiR-181c-5p internalized by microglia cells and inhibit the secretion of inflammation factors. ExomiR-181c-5p intrathecal administration alleviated neuropathic pain and neuroinflammation response in CCI rats. Taken together, ExomiR-181c-5p alleviated CCI-induced NP by inhibiting neuropathic inflammation. ExomiR-181c-5p may be a valid alternative for the treatment of neuropathic pain and has vast potential for future development.
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Exosomas , MicroARNs , Neuralgia , Animales , Exosomas/metabolismo , Inflamación/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Neuralgia/genética , Neuralgia/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Anastomotic leakage (AL) is one of most severe postoperative complications following low anterior resection (LAR) for rectal cancer, and has an adverse impact on postoperative recovery. The occurence of AL is associated with several factors, while few studies explored the role of intracorporeal barbed suture reinforcement in it. METHODS: Consecutive cases underwent laparoscopic LAR for rectal cancer from Mar. 2018 to Feb. 2021 in our center were retrospectively collected. Cases were classified into the intracorporeal barbed suture reinforcement group and the control group according to whether performing intracorporeal reinforcement with barbed suture, and AL incidences were compared between two groups. Propensity score matching (PSM) was then performed based on identified risk factors to reduce biases from covariates between two groups. AL incidences in the matched cohort were compared. RESULTS: A total of 292 cases entered into the study, and AL incidences were significantly lower in the intracorporeal barbed suture reinforcement group compared with the control group (10.00% vs 2.82%, P = 0.024). Sex, BMI, preoperative adjuvant chemoradiotherapy and anastomotic level were chose for PSM analyses based on previous studies. In the matched cohort, the AL incidences were still significantly lower in the intracorporeal barbed suture reinforcement group (10.57% vs 2.44%, SD = 0.334). CONCLUSIONS: Intracorporeal barbed suture reinforcement is associated with low AL incidences after laparoscopic LAR for rectal cancer, which is a potential procedure for reducing AL and worthy of application clinically.
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Laparoscopía , Neoplasias del Recto , Fuga Anastomótica/epidemiología , Fuga Anastomótica/etiología , Fuga Anastomótica/prevención & control , Humanos , Laparoscopía/efectos adversos , Neoplasias del Recto/cirugía , Estudios Retrospectivos , SuturasRESUMEN
CONTEXT: Veratramine may have a potential therapeutic effect for diabetic peripheral neuropathy (DPN). OBJECTIVE: To evaluate whether veratramine ameliorates neuropathic pain in a rat diabetic model. MATERIALS AND METHODS: Sprague-Dawley rats were used for a diabetic model induced by a streptozotocin + high-fat diet. Two months after the induction of the diabetic model, the rats with DPN were screened according to the mechanical pain threshold. The rats with DPN were divided into a model group (n = 12) and a treated group (n = 12). Rats with diabetes, but without peripheral neuropathy, were used in the vehicle group (n = 9). The treatment group received 50 µg/kg veratramine via the tail vein once a day for 4 weeks. During modelling and treatment, rats in all three groups were fed a high-fat diet. RESULTS: The mechanical withdrawal threshold increased from 7.5 ± 1.9 N to 17.9 ± 2.6 N in DPN rats treated with veratramine. The tolerance time of the treated group to hot and cold ectopic pain increased from 11.8 ± 4.2 s and 3.4 ± 0.8 s to 20.4 ± 4.1 s and 5.9 ± 1.7 s, respectively. Veratramine effectively alleviated L4-L5 spinal cord and sciatic nerve pathological injury. Veratramine inhibited the expression of SIGMAR1 and the phosphorylation of the N-methyl-d-aspartate receptor (NMDAR) Ser896 site in spinal cord tissue, as well as inhibited the formation of SIGMAR1-NMDAR and NMDAR-CaMKII complexes. DISCUSSION AND CONCLUSIONS: Veratramine may alleviate the occurrence of pain symptoms in rats with DPN by inhibiting activation of the SIGMAR1-NMDAR pathway.
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Diabetes Mellitus , Neuropatías Diabéticas , Neuralgia , Traumatismos de los Nervios Periféricos , Animales , Ratas , Neuropatías Diabéticas/tratamiento farmacológico , Neuralgia/tratamiento farmacológico , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato/metabolismoRESUMEN
OBJECTIVE: This study employed a randomized controlled trial to assess the efficacy of virtual-reality (VR) simulators and physical model simulators on colonoscopy training to explore the optimal and evidence-based simulation training. DESIGN: Forty participants were divided into 2 groups and randomized as dyads: the VR simulator group and the physical model simulator group. All the participants performed a baseline test through porcine colonoscopy. After a 6 h simulation training, each participant underwent a post-test on a pig after bowel preparation, and the procedures were video-recorded. Both the baseline test and the post-test were blindly assessed by 2 experienced assistant director physicians based on the GAGES-C scoring system. SETTING: Simulation center, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai. PARTICIPANTS: Forty surgical residents without colonoscopy experience. RESULTS: Both the VR simulator group and the physical model simulator group improved significantly over the baseline test. The VR simulator group performed significantly better than the physical model simulator group, p=0.042. The participants in both groups expressed a high level of simulator satisfaction. CONCLUSIONS: Novice residents can benefit from both VR simulators and physical model simulators. The VR simulator was shown to be more effective for colonoscopy training. VR simulators were more recommended for novices conducting basic colonoscopy training.
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Competencia Clínica , Colonoscopía , Internado y Residencia , Entrenamiento Simulado , Realidad Virtual , Colonoscopía/educación , Humanos , Entrenamiento Simulado/métodos , Femenino , Masculino , Internado y Residencia/métodos , Porcinos , Adulto , Animales , Educación de Postgrado en Medicina/métodos , ChinaRESUMEN
PURPOSE: Growth differentiating Factor 15 (GDF15) is linked to several cancers, but its effect on chemoresistance in colorectal cancer (CRC) remains unclear. Here, we investigated the role of GDF15 in the chemotherapeutic response of CRC patients to oxaliplatin (L-OHP). METHODS: GDF15 levels in serum and tumour tissues were detected in CRC patients have received L-OHP-based neoadjuvant chemotherapy. The effects of GDF15 neutralization or GDF15 knockdown on cell proliferation, apoptosis and intracellular reactive oxygen species (ROS) levels were analysed in vitro and in vivo. Co-immunoprecipitation (Co-IP), Chromatin Immunoprecipitation (ChIP) and luciferase reporter assays were used to explore the interaction between GDF15 and Nrf2. RESULTS: In this study, we found that GDF15 alleviates oxidative stress to induce chemoresistance of L-OHP in CRC. Mechanically, GDF15 posttranscriptionally regulates protein stability of Nrf2 through the canonical PI3K/AKT/GSK3ß signaling pathway, and in turn, Nrf2 acts as a transcription factor to regulate GDF15 expression to form a positive feedback loop, resulting in the maintenance of redox homeostasis balance in CRC. Furthermore, a positive correlation between GDF15 and Nrf2 was observed in clinical CRC samples, and simultaneous overexpression of both GDF15 and Nrf2 was associated with poor prognosis in CRC patients treated with L-OHP. Simultaneous inhibition of both GDF15 and Nrf2 significantly increases the response to L-OHP in an L-OHP-resistant colorectal cancer cells-derived mouse xenograft model. CONCLUSION: This study identified a novel GDF15-Nrf2 positive feedback loop that drives L-OHP resistance and suggested that the GDF15-Nrf2 axis is a potential therapeutic target for the treatment of L-OHP-resistant CRC.
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Neoplasias Colorrectales , Resistencia a Antineoplásicos , Retroalimentación Fisiológica , Factor 15 de Diferenciación de Crecimiento , Homeostasis , Factor 2 Relacionado con NF-E2 , Oxaliplatino , Oxidación-Reducción , Factor 15 de Diferenciación de Crecimiento/metabolismo , Factor 15 de Diferenciación de Crecimiento/genética , Humanos , Oxaliplatino/farmacología , Oxaliplatino/uso terapéutico , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/tratamiento farmacológico , Factor 2 Relacionado con NF-E2/metabolismo , Resistencia a Antineoplásicos/efectos de los fármacos , Resistencia a Antineoplásicos/genética , Animales , Oxidación-Reducción/efectos de los fármacos , Retroalimentación Fisiológica/efectos de los fármacos , Línea Celular Tumoral , Homeostasis/efectos de los fármacos , Ratones Desnudos , Masculino , Ratones , Femenino , Transducción de Señal/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Persona de Mediana Edad , Ratones Endogámicos BALB C , Especies Reactivas de Oxígeno/metabolismo , Apoptosis/efectos de los fármacosRESUMEN
This study investigated the biological role and mechanism of circMETTL15 in colorectal cancer (CRC). Cancer tissues and matched adjacent normal tissues were collected. CircMETTL15, miR-374a-5p, and ESCO2 levels were detected by RT-qPCR and Western Blot. LoVo cells were selected for loss- and gain-of-function assays and rescue assays. Cell proliferation was detected by CCK-8 and colony formation tests, cell apoptosis and cell cycle were detected by flow cytometry, cell migration and invasion were detected by Transwell assay, and protein expression of ki-67, E-cadherin, N-cadherin, and cleaved caspase-3 was detected by Western blot. Through bioinformatics analysis and verification assays, the targeting relationship between circMETTL15, miR-374a-5p, and ESCO2 was studied. The results suggest that circMETTL15 was a stable circRNA that was highly expressed in CRC tissues and cells and was associated with tumor size, higher TNM staging, and lymph node metastasis in CRC patients. Functionally, knocking down circMETTL15 inhibited the proliferation, migration, invasion, and EMT of LoVo cells, and induced apoptosis. Overexpression of circMETTL15 showed the opposite effect. The effects of knockdown or overexpression of circMETTL15 on the biological behavior of LoVo cells were reversed by knockdown of miR-374a-5p or knockdown of ESCO2, respectively. Mechanistically, circMETTL15 acts as a ceRNA for miR-374a-5p to regulate ESCO2 expression, thereby promoting the biological behavior of LoVo cells. In conclusion, the results of this study reveal the role of circMETTL15 in CRC and the underlying molecular mechanism, which provides potential data support for the development of future CRC drugs.
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Neoplasias Colorrectales , MicroARNs , Humanos , Acetiltransferasas , Apoptosis/genética , Western Blotting , Proteínas Cromosómicas no Histona , Neoplasias Colorrectales/genética , MicroARNs/genética , ARN Circular/genéticaRESUMEN
This randomized controlled trial compared the efficacy of virtual-reality (VR) simulator training and surgical training on live pigs to explore the most effective and evidence-based training modality. Materials and methods: Thirty-six novice surgical residents without independent laparoscopic experience were randomly paired with a peer and randomized into three groups: VR simulator group (dyad training on LapSim VR simulators), pig surgery group (training on live, anesthetized pigs) and control group (training by a lecture on laparoscopic surgery, surgical videos and textbooks). After 6 h of training, all participants performed a simulated cholecystectomy procedure using a pig liver with adherent gallbladder working in pairs. All procedures were video-recorded and the recordings were saved on USB-sticks in a blinded fashion identifiable only by the unique participant number. All video-recordings were scored blindly and independently by two expert raters using the Global Operative Assessment of Laparoscopic Skills (GOALS) assessment instrument. Results: The performances in the three groups were significantly different, P less than 0.001. Both the VR simulation training group and the live pigs training group performed significantly better than the control group, both P values less than 0.001. However, there was no significant difference in the performance of the two simulation-based training groups, P=0.66. Conclusion: Novice surgical trainees can benefit from both VR simulator training and pig surgery simulation compared with traditional studying and there was no significant difference between the two modalities. The authors recommend that VR simulators should be used for basic training of laparoscopic skills and surgery on live animals should be reserved for higher-level surgical training.
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CONTEXT: Quercetin (que) is one abundant flavonol with a variety of biological activities. Previous studies have shown quercetin can reduce neuropathic pain in rats with chronic constriction injury (CCI). OBJECTIVE: To evaluate the effects of quercetin on neuropathic pain in CCI model and explore its underlying mechanism in vivo. MATERIALS AND METHODS: CCI model was established by ligating the sciatic nerve of right leg on the SD rats. They were divided into ten groups: sham group, CCI model, sham+ que, CCI+ que group (30, 60, 120 mg/kg), CCI+ AICAR, CCI+ que+ compound C, CCI+etoricoxib, and the control group. They were administered for 28 days, and were performed the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) during the experiment. At the end of the experiment, sciatic nerves and spinal cord segments of rats were collected, ELISA detected the expression of inflammatory factors, detected the microglia and astrocytes with fluorescence, and Western blot detected AMPK/MAPK pathway. RESULTS: Que could increase the MWT of CCI rats, improve the TWL of plantar, and reduce the inflammatory cells at the ligation site of the sciatic nerve. Also, que could reduce the levels of TNF-α, IL-6, and IL-1ß. Western blotting results showed that p-38 MAPK, p-ERK, and p-JNK were activated in the spinal dorsal horn of CCI model group. After treatment with que and AMPK agonists, the phosphorylation levels of related proteins were inhibited. In addition, the analgesic effect of que was abolished when the AMPK inhibitor was added. DISCUSSION AND CONCLUSION: Quercetin alleviated the inflammatory response of sciatic nerve and spinal dorsal horn in rats induced by CCI. Quercetin alleviates neuralgia in CCI rats by activating AMPK pathway and inhibiting MAPK pathway and its downstream targets, p-38, p-ERK, and p-JNK.
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BACKGROUND: Long non-coding RNAs (lncRNAs) display regulatory function flexibly in tumor onset and developments. Our study aimed to delve into the roles of lncRNA LINC01569 (LINC01569) in colorectal cancer (CRC) progression to study the potential mechanisms. METHODS: The genetic expression profiles of miR-381-3p and LINC01569 were measured by RT-PCR. The subcellular localization of LINC01569 in CRC cells was identified using subcellular fractionation location. Loss-of-function assays were performed to explore the potential effects of LINC01569 on CRC progression. Dual-luciferase reporter analysis was employed to verify the binding connections among LINC01569, miR-381-3p, and RAP2A. RESULTS: LINC01569 expression was distinctly increased in CRC. Curiously, if LINC01569 is removed, CRC cells will not migrate, proliferate, and invade remarkably. Molecular mechanism exploration uncovered that LINC01569 acted as a ceRNA competing with RAP2A to bind with miR-381-3p. Furthermore, rescue experiments corroborated the fact that miR-381-3p suppression reversed the inhibitory actions of LINC01569 knockdown on the expression of RAP2A and CRC progression. CONCLUSION: Overall, our findings indicate that LINC01569 plays a key role in CRC development by means of aiming at the miR-381-3p/RAP2A axis and can be equivalent to an underlying medicinal target to save CRC patients.
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Objective: To explore the potential regulation mechanisms of miR-384-5p in Neuropathic pain (NP).Methods: Rat model of chronic constriction injury (CCI) was established to induce NP in vivo. NP levels were assessed using Withdrawal Threshold (PWT) and Paw Withdrawal Latency (PWL). qPCR and Western blotting were used to determine the relative expression of miR-384-5p and SCN3A. The inflammation response in spinal microglia cells was determined by ELISA assay. Immunofluorescence assay was used to demonstrate the co-localization of miR-384-5p with SCN3A in rat dorsal root ganglions (DRGs). The target genes of miR-384-5p were verified by dual-luciferase report assays.Results: In the current study, the miR-384-5p expression level was significantly downregulated in CCI rats when comparing to the sham group. In addition, miR-384-5p agomir significantly repressed mechanical allodynia and heat hyperalgesia in CCI rats. Meanwhile, the current study indicated miR-384-5p could decrease inflammation progress in spinal microglia cells incubated in lipopolysaccharide. Consistently, overexpression of miR-384-5p obviously depressed inflammation cytokine levels in CCI rats. Dual-luciferase reporter assays indicated that SCN3A is a target gene of miR-384-5p.Conclusion: miR-384-5p is a negative regulator in the development of neuropathic pain by regulating SCN3A, indicating that miR-384-5p might be a promising therapeutic target in the treatment of neuropathic pain.Abbreviations: CCI: Chronic constriction injury; ZEB1: Zinc finger E box binding protein-1; MAPK6: Mitogen-activated protein kinase 6; COX-2: cyclooxygenase-2.
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Modelos Animales de Enfermedad , MicroARNs/biosíntesis , Canal de Sodio Activado por Voltaje NAV1.3/biosíntesis , Neuralgia/metabolismo , Neuralgia/prevención & control , Animales , Constricción , Células HEK293 , Humanos , Masculino , MicroARNs/genética , Canal de Sodio Activado por Voltaje NAV1.3/genética , Neuralgia/genética , Ratas , Ratas Sprague-DawleyRESUMEN
AIMS: Postoperative cognitive dysfunction (POCD) is a common postoperative complication that is associated with increased morbidity and mortality. However, the mechanism of pathogenesis of POCD still remains largely unknown. The aim of the study was to investigate the function and mechanism of lncRNA PCAI in POCD. MATERIALS AND METHODS: Knockdown and overexpression studies were performed to analyze the function of lncRNA PCAI in cultured BV-2 cell lines treated with LPS to mimic the neuroinflammation. Real-time PCR, western blot, ELISA were used to determine the expression level of inflammation markers. Rescue experiment was performed to prove the relationship between PCAI and SUZ12. RESULTS: We found that the expression of lncRNA PCAI was decreased with the increasing concentrations of LPS. Knockdown of lncRNA PCAI inhibited the cell death rates and attenuated the cell inflammation via ELISA and real-time PCR. Besides, downregulated of lncRNA PCAI can protect the mitochondrial function via membrane potential assay. Overexpression of lncRNA PCAI can promote the cell death and inflammation response induced by LPS. We also provided mechanism study about lncRNA PCAI that negatively regulating SUZ12. Rescue experiment also verified the results. CONCLUSION: We performed comprehensive study of functional analysis of lncRNA PCAI in POCD and proved its mechanism, which negatively regulate SUZ12. Our study provided new clues for the clinical intervention and targets for POCD.
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Disfunción Cognitiva/etiología , Hipocampo/metabolismo , Inflamación/metabolismo , Complicaciones Cognitivas Postoperatorias/prevención & control , Complicaciones Posoperatorias/prevención & control , ARN Largo no Codificante/genética , Animales , Línea Celular , Regulación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Inflamación/patología , Ratones , Membranas Mitocondriales/metabolismo , Proteínas de Neoplasias , Complejo Represivo Polycomb 2/genética , Sustancias Protectoras/metabolismo , Sustancias Protectoras/farmacología , Factores de TranscripciónRESUMEN
OBJECTIVE: Colorectal cancer (CRC) is highly malignant and cancer metastasis remains the predominant cause of CRC death. The potential molecular mechanism of long non-coding RNA (lncRNAs) in CRC malignance is still poorly elucidated. METHODS: CCMAlnc expression was analyzed by using the Sequence ReadArchive (SRA) database. Target gene expression was examined by real-time PCR and Western blotting. The biological function of CCMAlnc and miR-5001-5p was detected by cell invasion, CCK8 proliferation, and colony formation assays in loss of function and gain of function experiments in vitro. A luciferase assay was performed to validate the target site of miR-5001-5p on the 3'-UTR of HES6 mRNA. RESULTS: CCMAlnc was identified as a novel functional lncRNA in CRC. Elevated CCMAlnc was detected in CRC cells as well as in clinical CRC tissue samples, and the expression of this lncRNA positively correlated with the poor prognosis of CRC patients. Functional validation assays revealed that downregulation of CCMAlnc impaired CRC cell proliferation and invasion in vitro, but upregulation of CCMAlnc reversed this effect. Moreover, CCMAlnc was validated to act as a competing endogenous RNA (ceRNA) that stabilizes the expression of HES6 by downregulating miR-5001-5p. CONCLUSION: CCMAlnc/miR-5001-5p/HES6 signaling is strongly activated to promote CRC malignance. CCMAlnc is defined as a potential candidate biomarker for metastasis prediction in CRC patients and as a potential therapeutic target for CRC treatment.
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Neuropathic pain is rarely diagnosed. Oxyntomodulin is peripherally and centrally distributed; however, the potential mechanisms underlying the effects of oxyntomodulin in attenuating nociception remain unclear; thus, we aimed to explore them in the present study. A neuropathic pain model in male C57BL/6 mice was induced by intrathecal injection of tumor necrosis factorα (TNFα), and the duration of nociceptive behavioral responses was measured with a stopwatch timer within 30 min. Western blotting was used to explore the protein levels of ionized calcium binding adaptor molecule1 (IBA1), nuclear factorκB (NFκB) phosphorylatedp65, interleukin (IL)6 and IL1ß. We performed reverse transcriptionquantitative polymerase chain reaction and ELISA were performed to determine the mRNA and protein expression levels of IL6 and IL1ß, respectively. An MTT assay was conducted to detect BV2 cell viability. Oxyntomodulin was observed to attenuate TNFαinduced pain hypersensitivity in mice, as well as the expression of IBA1, NFκB pp65, IL6 and IL1ß in the spinal cord. Oxyntomodulin exhibited no cytotoxicity on BV2 cells, and attenuated TNFαinduced IL6 and IL1ß production and release in BV2 cells and culture medium, respectively. Collectively, we proposed oxyntomodulin to attenuate TNFα induced neuropathic pain associated with the release of glial cytokines IL6 and IL1ß via inhibiting the activation of the NFκB pathway.
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Susceptibilidad a Enfermedades , Neuralgia/etiología , Neuralgia/metabolismo , Oxintomodulina/metabolismo , Transducción de Señal , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Biomarcadores , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Modelos Animales de Enfermedad , Expresión Génica , Masculino , Ratones , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Microglía/efectos de los fármacos , Microglía/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Neuralgia/tratamiento farmacológico , Oxintomodulina/farmacología , Fosforilación , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Obesity is a major epigenetic cause for colorectal cancer (CRC). Leptin is implicated in obesity-associated CRC, but the underlying mechanism remains unclear. The current study identified over-expression of metallopanstimulin-1 (MPS-1) in CRC patients through microarray and histological analysis, especially in obese CRC patients. MPS-1 was correlated with advanced tumor stage, suggesting its association with CRC progression. In addition, MPS-1 over-expression was associated with poor overall survival (OS) in obese CRC patients, but not in their non-obese counterparts, suggesting its potential as a prognostic marker of obese CRC patients. MPS-1 expression was positively associated with circulating leptin levels in CRC patients, especially in obese cases. Functional experiments demonstrated that MPS-1 silencing inhibited tumor proliferation and colony formation, and induced apoptosis of CRC cells in vitro. Converse results were obtained from the experiments with MPS-1 over-expression. Mechanistically, MPS-1 executed its action through induction of c-Jun N-terminal kinase (JNK)/c-Jun pathway. Moreover, the promotion effect of MPS-1 on CRC progression was modulated by leptin. In vivo studies demonstrated that MPS-1 silencing suppressed tumor growth of CRC via inhibiting JNK/c-Jun signaling. Collectively, this study indicates that MPS-1 promotes leptin-induced CRC via activating JNK/c-Jun pathway. MPS-1 might represent a potent candidate for the treatment and prognostic prediction of obesity-associated CRC.
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Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , Leptina/metabolismo , Metaloproteínas/metabolismo , Proteínas Nucleares/metabolismo , Proteínas de Unión al ARN/metabolismo , Proteínas Ribosómicas/metabolismo , Transducción de Señal , Células CACO-2 , Neoplasias Colorrectales/patología , Femenino , Células HCT116 , Humanos , MAP Quinasa Quinasa 4/metabolismo , Masculino , Proteínas Proto-Oncogénicas c-jun/metabolismoRESUMEN
BACKGROUND: The long noncoding RNA (lncRNA) colon cancer-associated transcript-1 (CCAT1) has been reported to play a vital role in the development of cancer. Although the link between inflammation and cancer initiation is well established, whether CCAT1 is involved in inflammation and promotes inflammatory bowel disease (IBD) malignancy remains undetermined. We aimed to investigate the expression of CCAT1 in IBD and the effect of CCAT1 overexpression on intestinal epithelial barrier function. METHODS: The relationship between CCAT1 and the inflammation-related pathway was analyzed in both colorectal cancer (CRC) and IBD patients. Gene expression was detected by real-time polymerase chain reaction and Western blot. Transepithelial electrical resistance (TEER) and FD-4 flux measurement were used to test the effect of CCAT1 and miR-185-3p on intestinal epithelial barrier function. Luciferase assay was performed to validate the target site of miR-185-3p on 3'-UTR of MLCK mRNA. RESULTS: Gene set enrichment analysis revealed that several inflammation-related genes were enriched in the CCAT1 high-expressed group of CRC patients. The relationship between CCAT1 and inflammation activation in IBD patients was further confirmed. CCAT1 expression positively correlated with MLCK, which acts as a protein kinase to phosphorylate myosin light chain and induces tight junction protein distribution, whereas it was negatively correlated with miR-185-3p in IBD tissues. We also determined that CCAT1 overexpression increased Caco-2 monolayer permeability and upregulated MLCK. Furthermore, CCAT1-induced MLCK overexpression and IBD disease progression were significantly attenuated by miR-185-3p. CONCLUSIONS: The CCAT1/miR-185-3p/MLCK signaling pathway is strongly activated to destroy barrier function and promotes the pathogenesis of IBD.
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Permeabilidad de la Membrana Celular , Neoplasias Colorrectales/patología , Enfermedades Inflamatorias del Intestino/patología , Intestinos/patología , MicroARNs/genética , ARN Largo no Codificante/genética , Uniones Estrechas/patología , Células CACO-2 , Estudios de Casos y Controles , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Enfermedades Inflamatorias del Intestino/genética , Enfermedades Inflamatorias del Intestino/metabolismo , Quinasa de Cadena Ligera de Miosina/genética , Quinasa de Cadena Ligera de Miosina/metabolismo , Fosforilación , Transducción de Señal , Uniones Estrechas/genética , Uniones Estrechas/metabolismoRESUMEN
Ulcerative colitis (UC) is a chronic, non-specific inflammatory disease that occurs in the colonic mucosa. This study investigated the role of the Notch pathway in affecting the pathogenesis of UC and regulating intestinal epithelial cell proliferation and apoptosis. Caspase-3 activity was measured and flow cytometry was used to detect reactive oxygen species (ROS) content and Ki-67 expression. Flow cytometry was applied to detect apoptosis, proliferation, and ROS content. Under LPS stimulation conditions, the IEC-6 cells were divided into 3 groups, including control, 5 and 10 µg/mL Jagged-1 protein pretreatment. The mRNA and protein expressions of Jagged-1, Notch1, Hes1, and OLFM4 in colon tissues were detected by real-time quantitative PCR (qRT-PCR) and Western blot. The ROS production, Ki-67 expression, and caspase-3 activity were significantly increased, and Jagged-1, Notch1, Hes1, and OLFM4 mRNA and protein levels were obviously elevated in the colon tissue of UC model rats compared with control. LPS treatment apparently up-regulated Jagged-1, Notch1, and OLFM4 expression in IEC-6 cells, resulting in marked enhancement in apoptosis and ROS generation, and reduction of proliferation. Administration of Jagged-1 before LPS stimulation further upregulated the expressions of Notch1 and OLFM4 in IEC cells, weakened apoptosis and ROS production, and alleviated the inhibitory effect of LPS on IEC-6 cell proliferation. UC lesions can activate the Notch signaling pathway in colon tissue, which may play a role in emergency repair. Upregulation of the Notch signaling pathway significantly reduced inflammatory stimuli-induced apoptosis and ROS generation in intestinal epithelial cells, resulting in increased cell proliferation.
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OBJECTIVE: To study the effects of ultrasound guided inter-scalene brachial plexus block and patient-controlled infraclavicular brachial plexus block for postoperative pain and surgical efficacy in patients with terrible tyriad of the elbow. METHODS: From March 2015 to August 2016, 60 patients with terrible tyriad of the elbows were treated in Ningbo No.6 Hospital with ASA I to II internal fixation. There were 32 males and 28 females, ranging in age from 16 to 70 years old, with a mean age of (55.6±18.2) years old. All the patients were divided into two groups(30 cases in each group): controlled intermuscular groove brachial plexus block (group C), infraclavicular brachial plexus block(group I). All catheters were placed using ultra-sound visualization and injected 0.33% ropivacaine 30 ml preoperatively. After regaining consciousness, all patients connected the electronic pump. The solution contained 0.2% ropiva-caine and the pump was setup to deliver a 5 ml bolus dose, with a 15 min lock out interval and background infusion at 5 ml/h. Both analgesia lasted until 5 d after operation. The patients underwent rehabilitation exercise everyday for 5 consecutive days starting from 24 h after operation.VAS score was recorded at 24 h, 48 h, 72 h and 4 d, 5 d after operation during rest and rehabilitation exercise time. The elbow articular range of motion and Mayo elbow performance score (MEPS) were recorded at 6 d after operation. Catheter-related adversereactions (such as oozing from the insertion site, obstruction, prolapse) were recorded. RESULTS: The success rate of blockade was 100% during insertion in both groups. Compared with group C, the VAS score at 3 d during rest time and 3, 4, 5 d after operation during rehabili-tation exercise were decreased(2.5±0.5 vs. 3.8±1.1, 3.0±0.4 vs. 5.0±0.9, 2.5±0.4 vs. 4.5±1.2, 2.1±0.3 vs. 4.1±1.0, P<0.05). The elbow articular range of motion and MEPS were increased(-2.19±18.01)° vs.(-8.19±12.16)°, (45.15±11.20)° vs. (22.15±7.02)°, (19.06±6.75)° vs. (9.10±2.48)°, (17.08±5.18)° vs. (10.12±3.15)°, (80.80±9.50) points vs. (64.90±11.21) points. The incidence of insertion site, obstruction, prolapse was 15, 5 and 10 cases respectively in group C, but without any catheter-related adverse reactions happened in group I (P<0.05). CONCLUSIONS: Patient-controlled infraclavieular brachial plexus block can be effectively used for postoperative pain after fixation for terrible tyriad of the elbows, and it can increase surgical outcome.
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Analgesia Controlada por el Paciente/métodos , Bloqueo del Plexo Braquial/métodos , Luxaciones Articulares/cirugía , Dolor Postoperatorio/terapia , Fracturas del Radio/cirugía , Fracturas del Cúbito/cirugía , Adolescente , Adulto , Anciano , Anestésicos Locales , Clavícula , Articulación del Codo , Femenino , Humanos , Luxaciones Articulares/complicaciones , Masculino , Persona de Mediana Edad , Fracturas del Radio/complicaciones , Fracturas del Cúbito/complicaciones , Ultrasonografía Intervencional , Adulto JovenRESUMEN
Rab3D belongs to Rab protein family. Previous reports showed that the expression of Rab3D was dysregulated in various types of cancer. Rab3D belongsRab3D belongs. However, little is known about the role of Rab3D in carcinogenesis and progression of colorectal cancer (CRC). Here, we first evaluated the expression of Rab3D in 32 fresh CRC and matched normal tissues and found Rab3D was dramatically increased in CRC tissues compared to normal tissues (p<0.001). Furthermore, immunochemistry was used to investigate Rab3D expression in 300CRC tissue specimens. The expression of Rab3D significantly positively correlated with the tumor size (p=0.041), CEA level (p=0.007), tumor classification (p=0.030), lymphatic metastasis (p<0.001), distant metastasis (p=0.013) and clinical stage (p=0.003). We also demonstrated that overall survival is poor in CRC patients with high expression of Rab3D (p<0.001). Finally, we showed that Rab3D activated Akt/GSK3ß/Snail pathway and induced EMT process in colorectal cancer cells. In conclusion, this study establishes increased Rab3D expression is associated with invasiveness of CRC cells, and Rab3D expression status may serve as a reliable prognostic biomarker in CRC patients.
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Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteínas de Unión al GTP rab3/genética , Proteínas de Unión al GTP rab3/metabolismo , Anciano , Línea Celular Tumoral , Movimiento Celular , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Transición Epitelial-Mesenquimal , Femenino , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Pronóstico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Factores de Transcripción de la Familia Snail/metabolismo , Regulación hacia ArribaRESUMEN
OBJECTIVE: To determine whether lysosome-associated protein transmembrane-4 beta (LAPTM4B) over-expression is associated with the proliferation and invasion in colorectal cancer (CRC). METHODS: Thirty pairs of CRC tissues, containing carcinoma and adjacent tissues, were used for the examination of LAPTM4B mRNA expression by real-time quantitative PCR (qPCR) assays. Then immunohistochemistry was performed to examine LAPTM4B protein expression in 6 pairs of CRC tissues. Over-expression LAPTM4B and low-expression LAPTM4B cell models were constructed with HCT116 CRC cell lines. CCK8 assay was used to detect the proliferation and Transwell assay was used to detect the invasion of the model cells. RESULTS: qPCR and immunohistochemistry results showed that LAPTM4B expression levels in CRC were higher compared to adjacent tissues (all P<0.01). CCK8 and Transwell assays results showed that LAPTM4B promoted proliferation and invasion of HCT116 cell lines model cells (all P<0.01). CONCLUSION: LAPTM4B promotes the proliferation and invasion in CRC patients, and may be used as an important potential marker.
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Proliferación Celular , Neoplasias Colorrectales , Células HCT116 , Humanos , Inmunohistoquímica , Proteínas de la Membrana , Invasividad Neoplásica , Proteínas OncogénicasRESUMEN
ATPase family AAA domain-containing 2 (ATAD2) has been identified as a critical modulator involved in cell proliferation and invasion. The purpose of this study was to explore the expression of ATAD2 in CRC tissues as well as its relationship with degree of malignancy. Data containing three independent investigations from Oncomine database demonstrated that ATAD2 is overexpressed in CRC compared with normal tissue, and similar result was also found in 32 pairs of CRC tissues by qPCR. The protein expression of ATAD2 was examined in six CRC cell lines and 300 CRC specimens. The results showed that high expression of ATAD2 was significantly correlated with tumor size (P < 0.001), serum CEA (P = 0.012), lymph node metastasis (P = 0.018), liver metastasis (P = 0.025), and clinical stage (P = 0.004). Kaplan-Meier method suggested that higher ATAD2 protein expression significantly associated with the overall survival (OS) of CRC patients (P < 0.001) and was an independent predictor of poor OS. Functional studies showed that suppression of ATAD2 expression with siRNA could significantly inhibit the growth in SW480 and HCT116 cells. These results indicated that ATAD2 could serve as a prognostic marker and a therapeutic target for CRC.