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The system of hepatocyte growth factor (HGF) and its receptor c-Met plays a critical role in tumor invasive growth and metastasis. The mortality rate of colorectal cancer (CRC), one of the most commonly diagnosed malignancies, is increased by it gradual development into metastasis, most frequently in the liver. Overexpression of c-Met, the protein tyrosine kinase receptor for the HCF/scatter factor, has been implicated in the progression and metastasis of human colorectal carcinoma. In this study, we aimed to investigate the role of c-Met in CRC liver metastasis and illustrate the clinical impact of regulating HGF/c-Met signaling in patients with CRC liver metastasis. We found that (I) higher levels of c-Met expression (mRNA and Protein) in CRC liver metastasis than primary CRC by assessing the patient tissue samples; (II) a positive correlation of c-Met expression with tumor stages of CRC liver metastasis, as well as c-Met expression in CRC, live metastasis concurred with regional lymph node metastasis; (III) the clinical impact of downregulation of HGF/c-Met signaling on the reduction of proliferation and invasion in CRC liver metastasis. Therefore, we demonstrate that the regulation of HGF/c-Met pathways may be a promising strategy in the treatment of patients with CRC liver metastasis.
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Neoplasias Colorrectales , Regulación Neoplásica de la Expresión Génica , Factor de Crecimiento de Hepatocito/biosíntesis , Neoplasias Hepáticas , Hígado/metabolismo , Proteínas Proto-Oncogénicas c-met/biosíntesis , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/mortalidad , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/terapia , Femenino , Humanos , Hígado/patología , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidad , Neoplasias Hepáticas/secundario , Neoplasias Hepáticas/terapia , Metástasis Linfática , Masculino , Persona de Mediana Edad , Invasividad NeoplásicaRESUMEN
BACKGROUND: Addison's disease (AD) is a rare but potentially fatal disease in Western countries, which can easily be misdiagnosed at an early stage. Severe adrenal tuberculosis (TB) may lead to depression in patients. CASE SUMMARY: We report a case of primary adrenal insufficiency secondary to adrenal TB with TB in the lungs and skin in a 48-year-old woman. The patient was misdiagnosed with depression because of her depressed mood. She had hyperpigmentation of the skin, nails, mouth, and lips. The final diagnosis was adrenal TB that resulted in the insufficient secretion of adrenocortical hormone. Adrenocortical hormone test, skin biopsy, T cell spot test of TB, and adrenal computed tomography scan were used to confirm the diagnosis. The patient's condition improved after hormone replacement therapy and TB treatment. CONCLUSION: Given the current status of TB in high-burden countries, outpatient doctors should be aware of and pay attention to TB and understand the early symptoms of AD.
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Toxic-shock-syndrome toxin-1 (TSST-1), a superantigen, can stimulate T cell activation and be used for immunotherapy. In this study, we employed the carcinoembryonic antigen (CEA)-positive LoVo cells to test whether retrovirus-mediated TSST-1 expression could activate human T cells and promote cytotoxicity against tumor cells. We first generated plasmids of pLEGFP-N1-5HRE-CEAp-TSST-1-linker-CD80TM containing a fusion gene of the CEA promoter, 5 copies of the hypoxia-response elements (HRE) as an enhancer, the fragments for TSST-1, and the transmembrane domain of CD80 (CD80TM) and control pLEGFP-N1-5HRE-CEAp (without TSST-1) and generated retroviruses of 5HCTC and 5HC, respectively. After infection with 5HC and 5HCTC retroviruses to establish cell lines, the high levels of TSST-1 expression were observed on the membrane and cytoplasm of the 5HCTC-infected LoVo cells, particularly culture under a hypoxic condition, but not on CEA(-) HeLa cells. Furthermore, the TSST-1-expressing LoVo cell lysates, but not the control cell lysates, stimulated human T cell proliferation, and the co-culture of the TSST-1-expressing LoVo, but not control cells, with human peripheral blood mononuclear cells (PBMC) induced a high frequency of TNF-α- and IL-2-secreting T cells in vitro, particularly under hypoxic conditions. More importantly, co-culture of the TSST-1-expressing LoVo cells, particularly under hypoxic conditions, but not control cells, with different numbers of PBMC promoted potent cytotoxicity against LoVo cells in a dose-dependent manner in vitro. These data provide proof of the principle that selective induction of TSST1 expression in CEA(+) colorectal cancer (CRC) cells activates T cells that destroy tumor cells, particularly under a hypoxic condition. Therefore, our findings may aid in the design of new immunotherapy for the intervention of CRC at clinic.
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Toxinas Bacterianas/genética , Neoplasias Colorrectales/patología , Enterotoxinas/genética , Activación de Linfocitos/fisiología , Superantígenos/genética , Linfocitos T/inmunología , Antígeno B7-1/metabolismo , Toxinas Bacterianas/inmunología , Toxinas Bacterianas/metabolismo , Secuencia de Bases , Antígeno Carcinoembrionario/metabolismo , Hipoxia de la Célula/genética , Línea Celular Tumoral/virología , Proliferación Celular , Técnicas de Cocultivo , Neoplasias Colorrectales/inmunología , Pruebas Inmunológicas de Citotoxicidad/métodos , Relación Dosis-Respuesta Inmunológica , Enterotoxinas/inmunología , Enterotoxinas/metabolismo , Células HeLa , Humanos , Inmunoterapia/métodos , Interleucina-2/metabolismo , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Elementos de Respuesta/genética , Retroviridae/genética , Superantígenos/inmunología , Superantígenos/metabolismo , Linfocitos T/citología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The aim of the present study was to evaluate Fourier transform infrared spectroscopy (FTIR) monitoring of biochemical changes in apoptosis cells. Different concentrations of 5-fluorouracil (5-FU) treated colon cancer cell lines SW620 were used to determine the optimum concentration of 5-FU IC50 by means of MTT assay. Cell starvation and 5-Fu synergistic cell cycle arrest was in G1 and S phase. FTIR combined with flow cytometry was applied to analysis of SW 620 cells and SW620 cells treated with 5-FU for 12h, 24h (early apoptosis) and 48 h (late apoptosis) respectively. The peak position and the intensity of all bands were measured and comparison was made between the SW620 and apoptotic SW620 cells. Apoptosis cells have following characteristics compared with SW620 cells (1) The band at 1 740 cm-1 is an C=O stretching vibration. Changes in these bands can reflect lipid changes, and relative peak intensity ratio 11740/11460 significantly increased (p<0. 05), indicating that the relative contents of lipid in apoptosis cells increased. (2) The band at the 1 410 cm-1 peak represents that C-H stretching related was increased to amino acid residues and shifted to higher wave numbers compared to other groups. I1410o/I 460 at early and late death phase was significantly increased, which suggests that the relative contents of amino acid residues in apoptosis cells increased (p <0. 05). New vibrational bands at 1 120 cm-1 appeared at 24 h and increased at 48 h compared with other groups. The 1 120 cm-1 absorption band is mainly due to ser, serine and threonine C-O(H) stretching vibration, and I1120/I 1460 significantly increased (p<0. 05), indicating that the relative quantity of amino acid residues in apoptosis cells increased due to that DNA unwinds the double helix. (3) 1 240 cm-1 is mainly due to the asymmetric stretching modes of phosphodiester groups shifting to higher wave number, illustrating that nucleic acid conformation was changed in apoptosis cells. (4) The band 1 040 cm-1 associated with polysaccharide appeared at 24 and 48 h, meanwhile shifted to higher wave number, suggesting that polysaccharide decreased in late apoptotic cells, and I 1040/I1400 increased at late stage apoptosis, indicating that the relative content of polysaccharide in apoptosis cells increased. The authors' results suggest that FTIR applied to monitoring SW620 cells apoptosis may be as a potential diagnostic tool for cancer chemotherapy monitoring.
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Apoptosis , Fluorouracilo , Espectroscopía Infrarroja por Transformada de Fourier , Línea Celular Tumoral , Neoplasias del Colon/patología , HumanosRESUMEN
Sciatic hernia is one of the rarely pelvic floor hernias. We report a 45-year-old woman who presented with acute crampy pain of hypogastrium which radiated down the back of the left thigh and found a mass in her left buttock area which is about fist size with local pain, so she had to force to bow position when walking. She was also associated with definite gastro-intestinal symptoms. Computed tomography (CT) of the pelvis and abdomen demonstrated the herniation of an ileal loop through the sciatic foramen on the left side. The diagnosis and management of this case are herein described and previous publications on sciatic hernias are reviewed.
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Bile acids serve a critical role in the induction of gastric intestinal metaplasia (IM) and gastric carcinogenesis. The present study investigated the effects of bile acids on the induction of gastric IM formation. The results demonstrated that the expression levels of caudalrelated homeobox transcription factor 2 (CDX2), mucin 2 (MUC2) and farnesoid X receptor (FXR) were increased in vitro and in vivo following treatment with bile acids, and CDX2 transcriptionally activated MUC2 expression. Furthermore, knockdown of FXR attenuated bile acidenhanced CDX2 promoter activity and protein expression. Conversely, the FXR agonist GW4064 synergistically enhanced bile acidinduced CDX2 promoter activity. Bile acid treatment led to an increase in nuclear factor (NF)κB activity and protein expression. Treatment with GW4064 or the FXR antagonist Zguggulsterone enhanced or attenuated bile acidinduced NFκB activity, respectively. In addition, quantitative chromatin immunoprecipitation confirmed that bile acids led to enhanced binding of p50 to the CDX2 promoter, whereas this effect was not observed for p65. Treatment with GW4064 or Zguggulsterone enhanced and attenuated the binding activity of p50 to the CDX2 promoter, respectively. These results indicated that bile acids may activate the FXR/NFκB signalling pathway, thereby upregulating CDX2 and MUC2 expression in normal gastric epithelial cells.
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Ácidos y Sales Biliares/metabolismo , Mucosa Gástrica/patología , Metaplasia/etiología , Transducción de Señal/fisiología , Adulto , Anciano , Animales , Ácidos y Sales Biliares/farmacología , Factor de Transcripción CDX2/metabolismo , Línea Celular , Femenino , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/metabolismo , Humanos , Intestinos/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Mucina 2/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Regulación hacia ArribaRESUMEN
AIM: To characterize the expression of members of the transforming growth factor-beta (TGF-beta)/Smad/connective tissue growth factor (CTGF) signaling pathway in the tissue of benign biliary stricture, and to investigate the effect of TGF-beta signaling pathway in the pathogenesis of benign biliary stricture. METHODS: Paraffin embedded materials from 23 cases of benign biliary stricture were analyzed for members of the TGF-beta/Smad/CTGF signaling pathway. TGF-beta (1), TbetaR I , TbetaR II , Smad4, Smad7 and CTGF protein were detected by immunohistochemical strepto-advidinbiotin complex method, and CTGF mRNA was evaluated by hybridization in situ, while 6 cases of normal bile duct served as controls. The percentages of positive cells were counted. The correlation between TGF-beta (1), Smad4 and CTGF was analyzed. RESULTS: The positive expression ratios of TGF-beta (1), TbetaR I , TbetaR II , Smad4, CTGF and CTGF mRNA in 23 cases with benign biliary stricture were 91.3%, 82.6%, 87.0%, 78.3%, 82.6% and 65.2%, respectively, significantly higher than that in 6 cases of normal bile duct respectively (vs 33.3%, 16.7%, 50.0%, 33.3%, 50.0%, 16.7%, respectively, P < 0.05). The positive expression ratio of Smad7 in cases with benign biliary stricture was 70.0%, higher than that in normal bile duct, but this difference is not statistically significant 70.0% vs 50%, P > 0.05). There was a positive correlation between positive expression of TGF-beta (1), Smad4 and CTGF in cases with benign biliary stricture. CONCLUSION: The high expression of TGF-beta/Smad/CTGF signaling pathway plays an important role in the pathogenesis of benign biliary stricture.
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Enfermedades de los Conductos Biliares/metabolismo , Transducción de Señal , Factor de Crecimiento Transformador beta1/metabolismo , Enfermedades de los Conductos Biliares/etiología , Enfermedades de los Conductos Biliares/patología , Factor de Crecimiento del Tejido Conjuntivo , Constricción Patológica , Femenino , Humanos , Proteínas Inmediatas-Precoces/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Masculino , Persona de Mediana Edad , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/metabolismo , Receptor Tipo I de Factor de Crecimiento Transformador beta , Receptor Tipo II de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/metabolismo , Proteína Smad4/metabolismo , Proteína smad7/metabolismo , Factor de Crecimiento Transformador beta1/genéticaRESUMEN
Oxidative stress and inflammation are involved in the pathogenesis of atherosclerosis. Calcium channel blockers (CCBs) inhibit the development of atherosclerosis, although the underlying molecular basis has not been completely elucidated. The present study was designed to investigate the effects of felodipine, a CCB, on inflammation and oxidative stress in human umbilical vein endothelial cells (HUVECs) and to examine the underlying mechanisms of action. Oxidized lowdensity lipoprotein (oxLDL) was used to induce an inflammatory response in HUVECs. The effects of felodipine were investigated by measuring the content of nitric oxide (NO) and reactive oxygen species (ROS), the mRNA and protein levels of intercellular adhesion molecule 1 (ICAM1) and vascular cell adhesion protein 1 (VCAM1), and the mRNA levels of endothelial NO synthase (eNOS) and inducible NO synthase (iNOS), in addition to the adhesion ability of U937 cells to HUVECs. ROS and NO levels were significantly increased in HUVECs following 24h treatment with 25 mg/l oxLDL (P<0.01). The increase in ROS was reversed by treatment with felodipine. In addition, NO levels were increased following treatment with 1 µmol/l felodipine (P<0.05). The mRNA expression of ICAM1, VCAM1, eNOS and iNOS was increased (P<0.05). Administration of 0.1 µM felodipine significantly decreased the expression of ICAM1, VCAM1, and iNOS (P<0.05). The number of U937 cells adhered to oxLDLtreated HUVECs was significantly increased compared with control, which was reversed by felodipine (0.1 µM). In conclusion, felodipine was demonstrated to inhibit oxidative stress and inflammatory responses, suggesting that it may be used to treat atherosclerosis.
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Felodipino/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Inflamación/metabolismo , Lipoproteínas LDL/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Células Cultivadas , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inflamación/tratamiento farmacológico , Inflamación/genética , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico Sintasa de Tipo III/genética , Óxido Nítrico Sintasa de Tipo III/metabolismo , Molécula 1 de Adhesión Celular Vascular/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
OBJECTIVE: To describe the incidence and clinical characteristics of acute severe high-altitude diseases in indigenous Tibetans. METHODS: The medical records of indigenous Tibetan patients with high-altitude pulmonary edema (HAPE) and high-altitude cerebral edema (HACE), who were treated in this hospital from June of 1956 to June of 2005, were retrospectively reviewed. RESULTS: A total of 3 184 cases of high-altitude disease were recorded in this period. Twenty four patients (0.75%, 24/3 184), 21 with HAPE and 3 with HACE, were indigenous Tibetans. Risk factors or precipitating factors were found in all the 24 cases, including getting into even higher altitude, exertion, cold, and alcohol drinking. From clinical symptoms, physical signs and laboratory examinations, it was found that 9 cases were complicated with multi-organ dysfunction. CONCLUSION: Indigenous Tibetans who travel between the plateau and the plain or to even higher altitude can suffer from hypoxic injury, even acute severe high-altitude disease, which may be complicated by multi-organ dysfunction.
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Mal de Altura/diagnóstico , Mal de Altura/etnología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Altitud , Mal de Altura/epidemiología , Pueblo Asiatico , Edema Encefálico/epidemiología , Niño , Preescolar , China/epidemiología , Femenino , Humanos , Incidencia , Lactante , Masculino , Persona de Mediana Edad , Edema Pulmonar/epidemiología , Adulto JovenRESUMEN
Caudalrelated homeobox transcription factor 2 (CDX2) is a transcription factor, which is specifically expressed in the adult intestine. It is essential for the development and homeostasis of the intestinal epithelium and its functions as a tumor suppressor have been demonstrated in the adult colon. The present study aimed to examine the inhibitory effects of the overexpression of CDX2 on subcutaneouslytransplanted tumors, derived from LoVo colon cancer cells, in nude mice, and to provide experimental evidence for the biotherapy of colon cancer. A pEGFPC1CDX2 eukaryotic expression vector was transfected into the LoVo cells via lipofection, and LoVo cells stablyexpressing CDX2 (pEGFPC1CDX2 cells) were obtained using G418 selection. A nude mouse subcutaneouslytransplanted tumor model was established by inoculating the nude mice with the pEGFPC1CDX2 cells, and the effects of overexpression of CDX2 on transplanted tumor growth in the LoVo cells were observed. Western blotting results demonstrated that the protein expression of CDX2 in the LoVo cells was higher in the pEGFPC1CDX2 cell group, compared with that in the pEGFPC1 cell group and the untreated cell group. At 20 days postinoculation with either pEGFPC1CDX2 or pEGFPC1, the transplanted tumor masses were significantly lower in the pEGFPC1CDX2 group, compared with those in the pEGFPC1 and untreated groups. Immunohistochemistry revealed that the expression levels of CDX2 and matrix metalloproteinase2 (MMP2) were detected in each group, and the protein expression of CDX2 was increased in the tumor tissues from the nude mice in the pEGFPC1CDX2 group. However the expression of MMP2 was downregulated in the tumor tissues of the nude mice in the pEGFPC1CDX2 group. Taken together, these data suggested that pEGFPC1CDX2 cells exhibited suppressed tumor growth in vivo. Overexpression of CDX2 was observed in transplanted tumors in the pEGFPC1CDX2 group, and the gene expression of MMP2 was reduced. These results indicate that CDX2 inhibited the growth of colorectal tumor cells, possibly by downregulating the gene expression.
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Neoplasias Colorrectales/metabolismo , Proteínas de Homeodominio/metabolismo , Animales , Factor de Transcripción CDX2 , Línea Celular Tumoral , Neoplasias Colorrectales/patología , Expresión Génica , Regulación Neoplásica de la Expresión Génica , Proteínas de Homeodominio/genética , Humanos , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Ratones Desnudos , Trasplante de Neoplasias , Carga TumoralRESUMEN
Caudal-related homeobox protein 2 (CDX2), a tumor suppressor in the adult colon, is overexpressed under a non-cancer specific cytomegalovirus promoter in certain tumor cells; furthermore, non-specific expression of CDX2 may result in aberrant side effects in normal cells. The human telomerase reverse transcriptase (hTERT) promoter is active in the majority of cancer cells but not in normal cells. Hypoxia is a key feature of solid tumors, and targeted genes may be significantly upregulated by five copies of hypoxia-response elements (HREs) under hypoxic conditions. However, the effect of CDX2 overexpression, as controlled by five copies of HREs and the hTERT promoter, on human colorectal cancer (CRC) cell proliferation in vitro remains to be fully elucidated. In the current study, a recombinant lentivirus containing the CDX2 gene under the control of five HREs and the hTERT promoter was generated. An immunofluorescence assay was used to detect CDX2 expression by the 5 HhC lentivirus, whereas an MTT assay was used to detect the effects of CoCl2 on the viability of LoVo cells. Western blot analysis was conducted in order to determine the relative ratios of recombinant CDX2 protein to the internal control ß-actin, following 5 HhC/LoVo cell culture under normoxic and hypoxic conditions (100, 200, 300, 400 or 500 µmol/l CoCl2) for 24 h, then for 12, 24 or 36 h with the optimal concentration (300 µmol/l) of CoCl2. Reverse transcription polymerase chain reaction analysis was used to determine the transcription of recombinant CDX2 mRNA following culture of 5 HhC/LoVo cells under normoxic or hypoxic conditions. Finally, a cloning assay was used to detect the proliferative ability of 5 HhC/LoVo and 5 Hh cells. High CDX2 expression was observed in hTERT-positive LoVo cells under hypoxic conditions, an effect which was mimicked by treatment with CoCl2 to inhibit LoVo cell proliferation in vitro. High expression of CDX2 therefore provides a promising strategy for the development of novel targeted treatments and gene therapy for CRC.
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Vectores Genéticos/metabolismo , Proteínas de Homeodominio/metabolismo , Lentivirus/genética , Factor de Transcripción CDX2 , Hipoxia de la Célula , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cobalto/toxicidad , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Vectores Genéticos/genética , Proteínas de Homeodominio/genética , Humanos , Microscopía Fluorescente , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Elementos de Respuesta/genética , Telomerasa/genéticaRESUMEN
Colorectal cancer is one of the main malignant tumors threatening human health. Surgery plays a pivotal role in treating colorectal cancer. The present study aimed to compare the clinical effect in patients with rectal cancer undergoing laparoscopic versus open surgery by meta-analysis of the randomized controlled trials (RCTs) published in the past 20 years. The data showed that 14 RCTs comparing laparoscopic surgery with conventional open surgery for rectal cancer matched the selection criteria and reported on 2,114 subjects, of whom 1,111 underwent laparoscopic surgery and 1,003 underwent open surgery for rectal cancer. Blood loss (P<0.00001), days to passage of flatus (P=0.0003), first bowel movement (P=0.0006), fluids intake (P<0.00001), walking independently (P<0.00001), length of hospital duration (P=0.003) and the rate of wound infection (P=0.04) were all significantly reduced following laparoscopic surgery. The incidence of complications, such as ureteric injury (P=0.33), urinary retention (P=0.43), ileus (P=0.05), anastomotic leakage (P=0.09) and incisional hernia (P=0.88), were not significantly different between the two groups. There were no significant differences in lymph nodes harvested (P=0.88), length of specimen (P=0.60), circumferential resection margin (CRM) (P=0.86), regional recurrence ((P=0.08), port site or wound metastasis (P=0.67), distant metastasis (P=0.12), 3-year overall survival (OS) (P=0.42), 3-year disease-free survival (DFS) (P=0.44), 5-year OS (P=0.60) and 5-year DFS (P=0.70). Therefore, laparoscopy for the treatment of patients with rectal cancer has the advantage of recovery and the same complications and prognosis as laparotomy, which indicates that laparoscopy may provide a potential survival benefit for patients with rectal cancer.
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Tumor cell microenvironment defines cancer development, also in hepatocellular carcinoma (HCC). Hepatic stellate cells (HSCs) are believed to be the key contributors to tumor microenvironment in HCC, yet their precise role in cancer progression is still unclear. The aim of this study was to determine the effect of human HSCs on progression of HCC using a subcutaneous xenograft nude mouse model. Nude mice were stratified to receive subcutaneous injections of human HCC cell line HepG2 and human HSC line LX-2 (HepG2 + LX-2), HepG2 alone, LX-2 alone, or phosphate-buffered saline. Tumor growth was assessed by measuring tumor size. After 30 days, final tumor size, weight, and histology were assessed. Compared with mice that were only injected HepG2 cells, mice injected with HepG2 + LX-2 exhibited more rapid tumor growth, increased tumor size and weight, higher tumor cell numbers due to increased proliferation and reduced apoptosis, increased fibrotic bands containing LX-2 cells, and increased tumor angiogenesis. In conclusion, HSCs play a significant role in promotion of HCC growth.
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Carcinoma Hepatocelular/patología , Transformación Celular Neoplásica , Células Estrelladas Hepáticas/patología , Neoplasias Hepáticas/patología , Animales , Apoptosis , Carcinoma Hepatocelular/irrigación sanguínea , Línea Celular Tumoral , Proliferación Celular , Femenino , Células Hep G2 , Humanos , Neoplasias Hepáticas/irrigación sanguínea , Ratones , Ratones Desnudos , Neovascularización Patológica/patología , Microambiente TumoralRESUMEN
BACKGROUND: In order to understand the underlying mechanisms by which weight loss surgeries improve metabolic profiles in type 2 diabetes mellitus (T2DM) patients and to evaluate the relevance of the length of the common limb in modulating various aspects of metabolism, we performed regular duodenal-jejunal bypass (DJB) and long-limb DJB (LL-DJB) surgeries in Goto-Kakizaki (GK) rats and compared their effects on glycemic control. METHODS: Male GK rats at 12 weeks of age were used for this study. Body weight, food intake, fasting glucose, glucagon-like peptide-1 (GLP-1) level, glucose tolerance, insulin sensitivity, cholesterol and triglycerides levels, and fecal energy content were monitored for 26 weeks after the two types of surgeries. RESULTS: We performed systematic analyses on GK rats after DJB or long-limb surgeries. Both procedures prevented body weight gain, reduced blood glucose and lipid levels, increased GLP-1 levels, and led to better insulin sensitivity. In general, LL-DJB displayed better effects than DJB, except that both surgeries caused similar increase in GLP-1 levels. CONCLUSIONS: Both DJB and LL-DJB surgeries triggered beneficial effects in GK rats. LL-DJB showed better outcomes than DJB, which may be due to reduced food intake and higher fecal energy content. This indicates that the length of the common limb could influence metabolic profiles of surgery recipients.
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Cirugía Bariátrica/métodos , Diabetes Mellitus Experimental/cirugía , Duodeno/cirugía , Yeyuno/cirugía , Animales , Glucemia/metabolismo , Peso Corporal , Diabetes Mellitus Experimental/sangre , Péptido 1 Similar al Glucagón/metabolismo , Glucosa , Prueba de Tolerancia a la Glucosa , Resistencia a la Insulina , Masculino , RatasRESUMEN
We investigated the mechanism and effects of sorafenib on hepatic stellate cell (HSC) viability and in the liver tumor microenvironment. The expression of α-smooth muscle actin (α-SMA) was measured immunocytochemically in the LX2 cells treated with differing concentrations of sorafenib. Changes in the platelet-derived growth factor (PDGF)-BB and tumor growth factor (TGF)-ß1 concentrations were detected in the LX2 supernatant using an enzyme-linked immunosorbent assay (ELISA). Expressions of the extracellular signal-regulated kinase 1 (ERK1), ERK2, and Akt signaling pathways were measured using a western blot assay. The LX2 cells were cocultured with HepG2 cells for 24 h to observe their effects on HepG2 cell invasive ability. (1) After treatment with various concentrations of sorafenib for 12, 24, 36, or 48 h, MTT assay showed that the viability of the treated LX2 cells was lower than in the controls. (2) As sorafenib concentration and time of exposure increased, α-SMA expression became weaker in the treated cells. (3) The PDGF-BB and TGF-ß1 concentrations decreased with higher concentration, and longer exposures under the same sorafenib concentration. (4) The ERK1, ERK2, and Akt expressions were identical between the treated and the control groups, but their phosphorylated expression decreased with increased concentrations of sorafenib. (5) The invasive ability of the HepG2 cells induced by the LX2 gradually decreased as sorafenib concentrations increased. Sorafenib suppressed α-SMA expression, inhibited PDGF-dependent signaling pathways in HSCs, downregulated the PDGF-BB and TGF-ß1 expression in the HSCs supernatant, and restrained viability of the HSCs, resulting in suppressed proliferation and invasion in the HepG2 cells.
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Antineoplásicos/farmacología , Carcinoma Hepatocelular/patología , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/patología , Neoplasias Hepáticas/patología , Niacinamida/análogos & derivados , Compuestos de Fenilurea/farmacología , Microambiente Tumoral/efectos de los fármacos , Actinas/metabolismo , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Células Hep G2 , Células Estrelladas Hepáticas/metabolismo , Humanos , Invasividad Neoplásica , Niacinamida/farmacología , SorafenibRESUMEN
AIM: To construct and identify a recombinant lentiviral vector containing shRNA for human neuropilin-1 (NRP-1) gene. METHODS: Four shRNA targeting the NRP-1 mRNA were designed to construct the pGCSIL-RFP-shNRP1 lentivirus vectors. The positive clone was chosen and confirmed by PCR and DNA sequencing. 293T cells were cotransfected with pGCSIL-RFP-shNRP1, pHelper1.0 and pHelper 2.0 to package the lentivirus and the titer of the virus was tested. After lentivirus-shRNA and over-expression plasmid containing NRP-1 were transfected into 293T cells, Western blotting was used to determine the expression of Flag gene in order to observe the inhibited efficacy of relative NRP-1 expression. RESULTS: PCR analysis and DNA sequencing demonstrated that the shRNA sequence was consistent with the human NRP-1. The titer of the recombinant lentiviral vector was 1×10(9); Tu/mL. The relative expression of NRP-1 protein in the transfected cells significantly decreased after treated with lentiviral-shRNA. CONCLUSION: We have constructed successfully the effective recombinant lentiviral vector containing shRNA for human NRP-1 gene.
Asunto(s)
Lentivirus/genética , Neuropilina-1/genética , ARN Interferente Pequeño/genética , Secuencia de Bases , Vectores Genéticos , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Interferencia de ARNRESUMEN
AIM: To develop an affinity peptide that binds to gastric cancer used for the detection of early gastric cancer. METHODS: A peptide screen was performed by biopanning the PhD-12 phage display library, clearing non-specific binders against tumor-adjacent normal appearing gastric mucosa and obtaining selective binding against freshly harvested gastric cancer tissues. Tumor-targeted binding of selected peptides was confirmed by bound phage counts, enzyme-linked immunosorbent assay, competitive inhibition, fluorescence microscopy and semi-quantitative analysis on immunohistochemistry using different types of cancer tissues. RESULTS: Approximately 92.8% of the non-specific phage clones were subtracted from the original phage library after two rounds of biopanning against normal- appearing gastric mucosa. After the third round of positive screening, the peptide sequence AADNAKTKSFPV (AAD) appeared in 25% (12/48) of the analyzed phages. For the control peptide, these values were 6.8 ± 2.3, 5.1 ± 1.7, 3.5 ± 2.1, 4.6 ± 1.9 and 1.1 ± 0.5, respectively. The values for AAD peptide were statistically significant (P < 0.01) for gastric cancer as compared with other histological classifications and control peptide. CONCLUSION: A novel peptide is discovered to have a specific binding activity to gastric cancer, and can be used to distinguish neoplastic from normal gastric mucosa, demonstrating the potential for early cancer detection on endoscopy.
Asunto(s)
Biblioteca de Péptidos , Péptidos/análisis , Neoplasias Gástricas/química , Neoplasias Gástricas/diagnóstico , Unión Competitiva , Biopsia , Línea Celular Tumoral , Detección Precoz del Cáncer , Ensayo de Inmunoadsorción Enzimática , Humanos , InmunohistoquímicaRESUMEN
The homeobox gene, CDX2, plays a major role in development, especially in the gut, and also functions as a tumor suppressor in the adult colon. In the present study, we investigated the effects of CDX2 expression on the proliferation, migration, and apoptosis of the human colon cancer cell line, Lovo. Lovo cells exogenously expressing CDX2 exhibited no significant differences in the percentage of cells in G1- and S-phase or in apoptosis, as determined by flow cytometry. MTT assay also confirmed that CDX2 expression had no effect on proliferation in these cells. Interestingly, conditioned medium collected from CDX2-overexpressing Lovo cells showed a significant decrease in secretion of MMP-2 and the invasive potential of these cells was significantly inhibited. Collectively, these data suggest that CDX2 may play a critical role in the migration and metastasis of colon carcinoma and over-expression of CDX2 in colon cancer cells markedly inhibits invasion. Based on these results, exogenous expression of CDX2 might be a promising option in the treatment of colon carcinoma.