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Traditional lead-based primary explosives present challenges in application to micro-energetics-on-a-chip. It is highly desired but still remains challenging to design a primary explosive for the development of powerful yet safe energetic films. Copper-based azides (Cu(N3)2 or CuN3, CA) are expected to be ideal alternatives owing to their properties such as excellent device compatibility, excellent detonation performance, and low environmental pollution. However, the significantly high electrostatic sensitivity of CA limits its use in micro-electro-mechanical systems (MEMS). This study presents an in situ electrochemical approach to preparing and modifying a CA film with excellent electrostatic safety using a Cu chip. Herein, a CA film is prepared by employing Cu nanorod arrays as precursors. Next, polypyrrole (PPy) is directly coated on the surface of the CA materials to produce a CA@PPy composite energetic film using the electrochemical process. The results show that CuN3 is first generated and gradually oxidized to Cu(N3)2, essentially forming enclosed nest-like structures during electrochemical azidation. The microstructure and composition of the product can be regulated by varying the current density and reaction time, which leads to controllable heat output of the CA from 521 to 1948 J g-1. Notably, the composite energetic film exhibits excellent electrostatic sensitivity (2.69 mJ) owing to the excellent conductivity of PPy. Thus, this study offers novel ideas for the further advances of composite energetic materials and applications in MEMS explosive systems.
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OBJECTIVE: To explore the expression of microRNA-155 in hepatocellular carcinoma (HCC) and its contribution to recurrence and prognosis of HCC after liver transplantation (LT). METHODS: The expression levels of microRNA-155 in 100 HCC samples were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Kaplan-Meier and Cox proportional regression analyses were utilized to determine the association of microRNA-155 expression with patient survivals. RESULTS: The expression levels of microRNA-155 were higher in primary HCC patients with post-LT recurrence (n = 45, mean relative level = 14.94) than those with non-recurrence (n = 55, mean relative level = 4.70) (P = 0.001) and correlated with micro-vascular invasion of HCC tissue samples (P = 0.001). The patients with a higher expression of microRNA-155 had significantly worse recurrence-free survival (RFS: (21.5 ± 3.2) months, log rank P < 0.001) and overall survival (OS: (29.3 ± 3.2) months, log rank P < 0.001) than those with a lower expression of microRNA-155 (RFS: (50.8 ± 3.2) months; OS: (54.6 ± 3.5) months). Multivariate analysis revealed that a high expression of miR-155 was an independent prognostic predictor. CONCLUSION: MicroRNA-155 is over-expressed in primary HCC with tumor recurrence and may serve as a novel biomarker for tumor recurrence and survival of HCC patients after LT. The detection of microRNA-155 is of clinical significance in HCC.
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Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Transplante de Fígado , MicroRNAs/metabolismo , Biomarcadores Tumorais , Carcinoma Hepatocelular/cirurgia , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Hepáticas/cirurgia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , PrognósticoRESUMO
PURPOSE: High rate of perineal surgical site infection (SSI) is the most common complication following abdominoperineal resection (APR), especially for extralevator abdominoperineal excision (ELAPE). The purpose of this study was to investigate the effect of continuous negative pressure drainage combined with intermittent irrigation (CNPDCII) in the presacral space on the perineal SSI following laparoscopic ELAPE for low rectal cancer. PATIENTS AND METHODS: The clinical data of 99 patients with low rectal cancer who underwent laparoscopic ELAPE surgery were retrospectively analyzed. Among the 99 patients, 46 patients received CNPDCII and 53 patients received conventional drainage in the presacral space after ELAPE. Self-made irrigation drainage tube: took a silicone drainage tube, cut 3 side holes at every 2cm intervals at the front end, and fixed a flexible tube of an intravenous needle at the front end of the silicone drainage tube. The conventional drainage tube or self-made irrigation drainage tube was placed in the presacral space and poked out from the inside of the ischial tuberosity. The incidence of SSI and other perioperative indicators between the two groups was compared within 30 days after surgery. RESULTS: There was no statistical difference in clinicopathological features between the two groups of patients (p>0.05). A statistically lower rate of SSI was found in CNPDCII group (17.4%, 8/46) than the conventional drainage group (35.8%, 19/53). The drainage tube retention time (7.8±1.2 d VS 9.4±1.6 d) and the postoperative hospital stay (9.7±1.4 d VS 11.9±2.3 d) in CNPDCII group were significantly shortened than the conventional drainage group. There was no statistical difference in operating theatre time and intraoperative blood loss between the two groups. Multivariate analysis confirmed that CNPDCII was an independent protective factor for SSI after ELAPE. CONCLUSION: CNPDCII can effectively reduce the incidence of SSI following laparoscopic ELAPE, which is simple, safe and effective.
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BACKGROUND: Identifying the genes and signaling pathways related to chemoresistance might facilitate the development of novel therapeutic strategies for colon cancer. In this study, we aimed to investigate the biological functions and underlying mechanisms of action of miR-19b and NR3C1, as well as their effects on chemosensitivity to oxaliplatin and prognosis of colon cancer patients. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical staining were used to analyze the expression of miR-19b and NR3C1. Dual firefly luciferase reporter gene analysis was used to identify miR-19b target genes. Associations of miR-19b and NR3C1 with survival were estimated by the Kaplan-Meier method and Cox regression analyses. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometric analysis were used to measure cell viability, cytotoxicity, cell cycle phase, and apoptosis, respectively. The effect of miR-19b on cell proliferation was investigated in vivo. RESULTS: The miR-19b was overexpressed and NR3C1 was decreased in colon cancer tissue and cell lines (SW480 and DLD-1). The miR-19b inhibition and NR3C1 overexpression inhibited cell proliferation, and induced G1/S cell cycle blockade, apoptosis, and chemosensitivity to oxaliplatin in vitro. The miR-19b inhibition suppressed subcutaneous tumorigenesis in vivo. Increased miR-19b and decreased NR3C1 in colon cancer were correlated with poor prognosis. In addition, our results confirmed NR3C1 was directly targeted by miR-19b. Thus, miR-19b might inhibit apoptosis and enhance oxaliplatin chemoresistance via the PI3K/AKT/mTOR pathway. CONCLUSIONS: Our study revealed that miR-19b promotes cell survival and chemoresistance to oxaliplatin via the PI3K/AKT/mTOR pathway by downregulating NR3C1 in colon cancer. miR-19b and NR3C1 might be potential intervention targets for chemoresistance of colon cancer.
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PURPOSE: To develop a risk scoring system that can predict the incidence of anastomotic leakage after laparoscopic rectal cancer surgery. PATIENTS AND METHODS: The clinical data of 387 patients with rectal cancer who underwent laparoscopic low anterior resection were retrospectively collected. Univariable and multivariable logistic regression analyses were used to evaluate independent risk factors for postoperative anastomotic leakage. A simplified points system was then developed based on the corresponding regression coefficient ß of each risk factor. Receiver operating characteristic (ROC) analysis was used to evaluate the performance and the optimal cut-off value in predicting anastomotic leakage. The performance of the points system was then externally validated in an independent cohort of 192 patients based in another institution. RESULTS: Anastomotic leakage occurred in 36 of 387 patients with rectal cancer (9.30%). Logistic multivariable regression analysis showed that males, maximum tumor diameter (≥5cm), operation time (≥180min), preoperative chemoradiation, intraoperative blood transfusion and the anastomosis level from the anal verge (≤5cm) were independent risk factors for the incidence of anastomotic leakage. According to the scoring standard, the risk points of each patient were calculated. ROC analysis based on the risk points showed that the area under the curve (AUC) was 0.795 (95% CI:0.752-0.834) and the optimal cut-off value was 6, yielding a sensitivity of 88.89% and a specificity of 62.96%. Using this risk points system, the AUC of another cohort of 192 patients from another institution who underwent laparoscopic low anterior resection for rectal cancer was 0.853 (95% CI:0.794-0.900, p<0.001) and patients with risk points ≥6 had a 21.05% chance of developing anastomotic leakage. CONCLUSION: This risk points system for predicting anastomotic leakage following laparoscopic rectal cancer surgery may be useful for surgeons in their decisions to perform intraoperative diversion stoma, which can reduce the incidence of postoperative anastomotic leakage.
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Background: There are few studies on postoperative complications after colonoscopic perforation. We aimed to study clinical characteristics and treatment after colonoscopic perforation, and to determine risk factors for postoperative complications by surgical treatment of colonoscopy perforation. Methods: Cases with perforation within 7 days after colonoscopy from January 2017 to December 2019 were collected for retrospective analysis. Data regarding demography, clinical information, colonoscopy, perforation, and operation were collected. Single-factor analysis and Spearman correlation analysis were employed to determine the risk factors of postoperative complications. Results: A total of 35,243 colonoscopy examinations were performed during the study period, of which 18 cases of colonoscopic perforation were included in the criteria. Most perforations occurred in the rectosigmoid junction (3 cases) and sigmoid colon (11 cases). All perforation patients received operational treatment, and the incidence of postoperative complications was 38.9%, but no deaths. There were 7 patients who developed postoperative complications. Spearman correlation analysis showed that preoperative medication of glucocorticoid and nonrectosigmoid perforation were positively related to postoperative complications (P < .05), while perforation diagnosed immediately and satisfying intestinal cleanliness were negatively related to it (P < .05). Conclusion: Perforation is a rare but serious complication of colonoscopy, which mostly occurs in the rectosigmoid junction and sigmoid colon. Laparoscopic primary repair is safe and feasible in resolving colonic perforation due to colonoscopy, and postoperative complications were significantly related to perforation site, preoperative medication of glucocorticoid, perforation diagnosis time, and intestinal cleanliness.
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Colo Sigmoide/cirurgia , Doenças do Colo/cirurgia , Colonoscopia/efeitos adversos , Perfuração Intestinal/etiologia , Adulto , Idoso , Endoscopia , Feminino , Glucocorticoides/uso terapêutico , Humanos , Incidência , Intestinos/cirurgia , Laparoscopia/efeitos adversos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Reto , Estudos Retrospectivos , Fatores de RiscoRESUMO
BACKGROUND: As a disease with extremely complex molecular mechanisms, many deregulated miRNAs have been identified in colon cancer. Few studies have been performed by using Ingenuity Pathways Analysis (IPA) to predict miRNAs specifically expressed in colon cancer. METHODS: A characteristic microRNA-target network of colon cancer was explored using IPA. Then the clinical significance of miR-19b-3p was evaluated in 211 colon cancer patients. The roles of miR-19b-3p and its candidate target gene, SMAD4, in colon cancer progression were examined both in vitro and in vivo. RESULTS: Bioinformatics analysis showed that 15 microRNAs screened by IPA were significantly correlated with malignant biological behaviors of colon cancer. miR-19b-3p was the most significantly upregulated candidate based on the validation experiment using 211 colon cancer samples. High expression of miR-19b-3p was significantly associated with high N stage (P < 0.001), high AJCC stage (P < 0.001), poor histologic grade (P = 0.032), frequent venous and lymphatic invasion (P = 0.027), and liver metastasis (P < 0.001). Survival analysis revealed that miR-19b-3p was an independent prognostic factor associated with colon cancer patient's overall survival (OS) and disease-free survival (DFS). miR-19b-3p promoted proliferation and chemoresistance of colon cancer cells, but had no effect on invasion in vitro, along with tumorigenesis in vivo. In addition, we confirmed that miR-19b-3p mediates resistance to oxaliplatin-based chemotherapy via SMAD4. CONCLUSIONS: Our findings demonstrate the role of miR-19b-3p-SMAD4 axis in colon cancer progression, which may become a potential therapeutic target against chemotherapy resistance.
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Neoplasias Colorretais/patologia , Biologia Computacional/métodos , Resistencia a Medicamentos Antineoplásicos , MicroRNAs/genética , Proteína Smad4/genética , Animais , Células CACO-2 , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias Colorretais/genética , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Células HCT116 , Células HT29 , Humanos , Masculino , Camundongos , Estadiamento de Neoplasias , Transplante de Neoplasias , Compostos Organoplatínicos/farmacologia , Oxaliplatina , Regulação para CimaRESUMO
In this study, we aimed to elucidate the clinical significance and underlying mechanisms of BRG1 in colon cancer. In the clinical analysis, overexpression of BRG1 correlates with colon cancer progression in two cohorts (n = 191 and n = 75). Kaplan-Meier survival analysis revealed that BRG1 is a prognosis predictor for overall survival (P < 0.001) and disease-free survival (P = 0.001). Knocking down BRG1 expression significantly suppressed the proliferation and invasion in colon cancer cells. The expression pattern of WNT3A is consistent with BRG1 in colon cancer tissues and WNT3A expression was inhibited in BRG1 knockdown cells. In addition, restoring WNT3A expression rescues the inhibition of cell proliferation and invasion induced by BRG1. In this study, we demonstrate that BRG1 may contribute to colon cancer progression through upregulating WNT3A expression.
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Neoplasias do Colo/etiologia , DNA Helicases/fisiologia , Proteínas Nucleares/fisiologia , Fatores de Transcrição/fisiologia , Proteína Wnt3A/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias do Colo/mortalidade , Neoplasias do Colo/patologia , Progressão da Doença , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Invasividade Neoplásica , PrognósticoRESUMO
BACKGROUND: The aim of this research was to investigate the impact of post-transplantation adjuvant chemotherapy in the prevention of tumor recurrence and metastasis for hepatocellular carcinoma (HCC) exceeding Milan criteria after liver transplantation. METHODS: A total of 117 patients with HCC exceeding the Milan criteria who had undergone orthotopic liver transplantation (OLT) from August 2002 to February 2009 were enrolled and retrospectively analyzed. The patients were divided into four groups according to chemotherapy regimens and the impact of different chemotherapy regimens on survival, disease-free survival, and adverse effects were compared. RESULTS: One year survival rates for the gemicitabine, conventional chemotherapy, oxaliplatin plus capecitabine and the best supportive care (BSC) group were 87.5%, 84.2%, 81.6%, and 67.5%. The 3-year survival rates were 48.1%, 25.9%, 31.6%, and 33.7%, respectively for the four groups. One year disease free survival rates for the four groups were 69.8%, 47.4%, 53.8%, and 45.7% respectively. And 3-year disease free survival rates were 43.2%, 23.7%, 23.6%, and 25.1% for the four groups. Stratification analysis showed that the gemcitabine regimen and conventional chemotherapy could significantly improve the survival rate and disease free survival rate for HCC patients who had major vascular invasion and/or microvascular invasion after liver transplantation compared with BSC group. CONCLUSIONS: For HCC patients beyond Milan criteria, especially who had vascular invasion and/or micorvascular invasion, post-transplantation adjuvant chemotherapy can significantly improve survival. Gemcitabine is a proper regimen for postoperative adjuvant chemotherapy. Conventional chemotherapy can also benefit patients, but the adverse effects are not satisfactory.
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Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/cirurgia , Quimioterapia Adjuvante/métodos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/cirurgia , Transplante de Fígado , Antineoplásicos/efeitos adversos , Antineoplásicos/uso terapêutico , Desoxicitidina/efeitos adversos , Desoxicitidina/análogos & derivados , Desoxicitidina/uso terapêutico , Doxorrubicina/efeitos adversos , Doxorrubicina/uso terapêutico , Feminino , Humanos , Masculino , GencitabinaRESUMO
The liver transplant (LT) situation represents an attractive model for studying hepatocellular carcinoma (HCC) metastasis. Based on microarray data, we previously found that miR-126 expression was lower in tumor tissues of patients with post-LT HCC recurrence compared with non-recurrence. In this study, we examined the expression of miR-126 in HCC samples from 68 patients who had undergone LT using quantitative real-time PCR and analyzed its correlation with clinicopathological features and prognosis of patients. Furthermore, we performed experimental analyses to explore the involvement of miR-126 in HCC metastasis. We found that miR-126 levels were lower in tumor tissues of patients with post-LT HCC recurrence in comparison to patients with no-recurrence (p = 0.009). Lower expression of miR-126 in HCC was associated significantly with tumor recurrence (p = 0.011) and poor survival (p = 0.009) of patients. Functional studies indicated that ectopic expression of miR-126 significantly inhibits HCC cells migration, invasion, proliferation and colony formation in vitro, and suppresses experimental lung colonization in vivo. Our study revealed that down-regulation of miR-126 plays an important role in HCC metastasis, and suggest a potential application of miR-126 in prognosis prediction and HCC treatment.
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Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Metástase Neoplásica , Adulto , Idoso , Carcinoma Hepatocelular/genética , Linhagem Celular Tumoral , Regulação para Baixo , Feminino , Humanos , Neoplasias Hepáticas/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , RecidivaRESUMO
PURPOSE: MicroRNAs play important roles in cancer development, progression, and metastasis. The aim of this study was to determine whether altered microRNA-155 expression is associated with hepatocellular carcinoma (HCC) recurrence and prognosis following orthotopic liver transplantation (OLT). METHODS: Tissue specimens from 100 HCC patients following OLT were recruited. MicroRNA-155 expression levels were detected using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Kaplan-Meier and Cox proportional regression analyses were utilized to determine the association of microRNA-155 expression with survival of patients. MicroRNA-155 expression levels of two HCC cell lines (HepG2 and SMMC-7721) and normal liver tissue were quantified using qRT-PCR. The potential function of miR-155 on invasiveness was evaluated in the above HCC cell lines. RESULTS: We found that microRNA-155 expression levels were high in tumor tissues in patients with post-OLT HCC recurrence (n = 45) compared with those in patients with non-recurrence (n = 55) (P = 0.001) and correlated with micro-vascular invasion of HCC tissue samples (P = 0.001). Patients with higher miR-155 expression had significantly poorer recurrence-free survival (RFS, log rank P < 0.001) and overall survival (OS, log rank P < 0.001). Multivariate analysis revealed that high miR-155 expression was an independent predictor of poor prognosis (HR 2.748, P = 0.001 for RFS; HR 5.752, P < 0.001 for OS). In addition, the invasiveness of HCC cells was significantly increased by higher microRNA-155 expression. CONCLUSIONS: MicroRNA-155 is a candidate oncogenic microRNA and plays an important role in promoting HCC cells invasion. Our findings suggest that microRNA-155 may serve as a novel biomarker for tumor recurrence and survival of HCC patients following OLT.
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Biomarcadores Tumorais/biossíntese , Carcinoma Hepatocelular/cirurgia , Neoplasias Hepáticas/cirurgia , Transplante de Fígado , MicroRNAs/biossíntese , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/irrigação sanguínea , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Hepáticas/irrigação sanguínea , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica , Recidiva Local de Neoplasia/genética , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Transfecção , Regulação para CimaRESUMO
Many recent studies have shown the utility of microRNAs (miRs) as cancer-related biomarkers. The aim of the present study was to evaluate the correlation between miR-203 expression and prognosis of patients with hepatocellular carcinoma (HCC) and cirrhosis after liver transplantation (LT). Sixty-six HCC samples from patients who had undergone LT were examined for miR-203 expression using quantitative reverse transcription-polymerase chain reaction. The data were correlated with clinicopathological parameters and prognosis. Patient survival was analyzed by the Kaplan-Meier method and log-rank test. Cox regression was used for multivariate analysis of prognostic factors. We found that miR-203 expression was low in tumor tissues of patients (n = 16) with post-LT HCC recurrence in comparison with those in patients with non-recurrence (n = 50) (P = 0.003). Patients with higher miR-203 expression had significantly better recurrence-free survival (RFS) and overall survival (OS) (P = 0.016 for RFS; P = 0.014 for OS). Multivariate analysis revealed that high-miR-203 expression was an independent predictor of good prognosis (HR 0.202, P = 0.006 for RFS; HR 0.332, P = 0.013 for OS). Our results suggest that miR-203 could be a novel prognostic marker in HCC patients who have undergone LT and might also be a potential therapeutic target.
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Carcinoma Hepatocelular/cirurgia , Cirrose Hepática/cirurgia , Transplante de Fígado/efeitos adversos , MicroRNAs/genética , Recidiva Local de Neoplasia/genética , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Cirrose Hepática/genética , Cirrose Hepática/patologia , Transplante de Fígado/mortalidade , Masculino , MicroRNAs/biossíntese , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Prognóstico , Modelos de Riscos Proporcionais , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Tumor recurrence-related microRNAs (miRNAs) in hepatocellular carcinoma (HCC) following orthotopic liver transplantation (OLT) are not clear yet. This study was designed to determine whether altered miRNA expression is associated with HCC recurrence and prognosis following OLT. 18 miRNAs, including 6 up-regulated and 12 down-regulated miRNAs were identified by microarray in primary HCC samples of patients who had developed HCC recurrence (n = 5) compared to those with non-recurrence (n = 5) following OLT by using p < 0.05 as cutoff value. The six most significantly altered miRNAs (fold change ≥ 2: miR-19a, miR-886-5p, miR-126, miR-223, miR-24 and miR-147) were further confirmed by qRT-PCR in the remaining 105 HCC samples. In receiver-operating characteristic curve analysis, this six miRNAs were of high sensitivity and specificity in predicting HCC recurrence. Using Cox regression and risk score analysis, we built a six-miRNA signature based on their qRT-PCR readings for the prediction of outcome of HCC following OLT. Kaplan-Meier and Cox proportional regression revealed this six-miRNA signature was a significant independent predictor of overall survival (log-rank p = 0.020) and recurrence-free survival (log-rank p < 0.001). Finally, the data were further reconfirmed in an independent cohort of 50 patients from another transplant center. In addition, bioinformatics Gene Ontology and pathway analysis were also performed to better understand the critical roles of these miRNAs in HCC recurrence. Our study, in addition to suggesting a different miRNA expression pattern between HCC samples of patients with recurrence and those with non-recurrence, proposes that this six-miRNA signature may serve as biomarker for prognosis of HCC patients following OLT.
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Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , Transplante de Fígado , MicroRNAs/genética , Recidiva Local de Neoplasia/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/patologia , Biologia Computacional , Feminino , Perfilação da Expressão Gênica , Redes Reguladoras de Genes/genética , Humanos , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/patologia , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Modelos de Riscos Proporcionais , Curva ROC , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/genética , Análise de SobrevidaRESUMO
The aim of this study was to detect the expression and cell cycle specificity of Fas, TNFRI and TNFRII in human peripheral blood lymphocytes (PBL), and to study the potential role of Fas, TNFRIand TNFRII in cell cycle specific apoptosis. The improved double-parameter flow cytometry was used to detect the expressions of Fas, TNFRI and TNFRII and cell cycle specificity in PBL which were incubated for 24 hours in the presence or absence of phytohaematoagglutinin (PHA) respectively. Apoptosis induced by IgM type anti-Fas and TNF-alpha was detected by API method. The results showed that compared with PBL treated in the absence of PHA in G(0) phase, the ratio of Fas, TNFRI and TNFRII expressions in PHA-stimulated PBL entering cell cycle increased (35.55 +/- 6.63)%, (30.63 +/- 2.66)%, (26.62 +/- 5.14)% respectively (P < 0.01), and mainly appeared at G(1)-phase; no apoptosis was induced by anti-Fas and TNF-alpha in G(0)-phase PBL cultured in the absence of PHA. On the contrary, the apoptosis was induced by anti-Fas and TNF-alpha in PBL which entered cell cycle after stimulation with PHA and mainly initiated at G(1)-Phase. It is concluded that there is evident dose-effect relationship between apoptotic receptor and receptor-mediated apoptosis. Moreover, the cell cycle specificity of receptor-mediated apoptosis is correlated with the cell cycle specific expressions of apoptotic receptor. The induction of apoptosis by apoptotic factors (anti-Fas and TNF-alpha) depends on whether cell entering cell cycle or not.
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Apoptose , Linfócitos/citologia , Receptores Tipo II do Fator de Necrose Tumoral/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Receptor fas/metabolismo , Ciclo Celular , HumanosRESUMO
BACKGROUND & OBJECTIVE: Cigarette smoke exposure has been reported to induce DNA damage in a variety of cell types. This study was to investigate DNA damage and apoptosis in normal human bronchial epithelial cell line NHBE and lung carcinoma cell line SPC-A1 caused by cigarette smoke extract. METHODS: NHBE and SPC-A1 cells were incubated with different concentrations of cigarette smoke extract. Cell viability was evaluated by MTT assay. Fluorescence-labeled anti-histone gamma-H2AX polyclonal antibody was used to detect DNA double-strand breaks (DSBS) in chromatin. DNA damage was analyzed by flow cytometry. The expression of gamma-H2AX was detected by Western blot. Cigarette smoke extract-induced cell apoptosis was detected by sub G1 peak method and annexin V-FITC/propidium iodide (PI) staining assay. Cell morphology of DNA damage and apoptosis was observed by confocal laser microscopy. RESULTS: MTT assay results showed that cigarette smoke extract decreased the viability of NHBE and SPC-A1 cells in time-and concentration-dependent manners. Cigarette smoke extract induced DSBS in NHBE cells and SPC-A1 cells, and led to H2AX phosphorylation (denoted gamma-H2AX) in time-and concentration-dependent manners. The maximal value of gamma-H2AX was seen about 4 h after the treatment, and the value decreased subsequently, but did not reduce to a normal level. Cell apoptosis appeared about 12 h after DNA damage. Cigarette smoke extract also initiated the accumulation of gamma-H2AX in these cells, and cell apoptosis morphology was observed 4 h later. CONCLUSION: Cigarette smoke extract can induce DSBS and apoptosis in NHBE and SPC-A1 cells in time-and concentration-dependent manners.
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Apoptose/efeitos dos fármacos , Dano ao DNA , Células Epiteliais/citologia , Nicotiana , Fumaça/efeitos adversos , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Brônquios/citologia , Linhagem Celular , Linhagem Celular Tumoral , Quebras de DNA de Cadeia Dupla , Relação Dose-Resposta a Droga , Células Epiteliais/metabolismo , Histonas/metabolismo , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Fosforilação , Fumaça/análise , Fatores de TempoRESUMO
BACKGROUND & OBJECTIVE: Autophagy is the main phenomenon of type II programmed cell death which is also named as autophagic cell death, and autophagy has a close relationship with autophagic cell death. The relationship of apoptosis and cell cycle has been explored deeply, but little is known about the relationship of autophagic cell death and cell cycle. This study was to observe the correlation of autophagy induced by different methods to cell cycle. METHODS: Exponentially growing HeLa and SW480 cells, and peripheral blood lymphocytes (PBLs) from healthy donors, with or without 48 h stimulation of phytohemagglutinin (PHA), were treated with Hanks' solution (to produce starvation) or vincristine. Confocal laser microscope and transmission electron microscope (TEM) were used to detect autophagy; flow cytometry (FCM) was innovatively used to detect the cell cycle of autophagic cells with dipl-parameters of microtubule-associated protein 1 light chain 3 (MAP1-LC3-II)/PI. RESULTS: Autophagy of HeLa and SW480 cells induced by starvation or vincristine was observed in G1, S, and G2/M phases and increased along with the inducement time; no autophagy was observed in unstimulated PBLs. The positive rate of LC3-II, indicating the occurrence of autophagy, was lower than 2.62% when induced by starvation in Hanks' solution for 48 h, or 6.16% when induced by vincristine for 48 h. After PBLs were stimulated into cell cycle by PHA, autophagy was markedly detected 2 h after the indicated inducements. CONCLUSIONS: MAP1-LC3-II/DNA dipl-parameter analysis by FCM is a convenient and reliable method for simultaneously analyzing autophagy and cell cycle. Autophagy could be induced when cells are in cell cycle, while the cells in G0 phase are insensitive to the inducers.