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OBJECTIVES: It is not clear whether being overweight or obese influences postoperative complications in myasthenia gravis (MG) patients. We retrospectively investigated an association between body mass index (BMI) and postoperative complications in MG. MATERIALS AND METHODS: Fifty-nine MG patients who had undergone transsternal thymectomy were classified as low or high BMI based on the criteria for Asian-Pacific populations. An association between BMI and complications was analyzed. RESULTS: MG patients with high BMI had significantly higher rates of major adverse complications (P = 0.033), postoperative respiratory failure (P = 0.045), and longer postoperative hospitalization (P = 0.005). The optimal cutoff value of BMI for postoperative respiratory failure was 23.3 kg/m2, with a sensitivity of 75.0% and a specificity of 64.7% (P = 0.046). CONCLUSIONS: MG patients with a BMI indicating overweight or obesity have a higher risk of postoperative complications after thymectomy. Thus, close monitoring must be performed when surgery is necessary.
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OBJECTIVE: We aimed to investigate how the embryonic stem cell-related gene Oct3/4 changes during the injury-repair process of distal pulmonary epithelium induced by 5-fluorouracil (5-Fu). METHODS: We have developed the lung injury model induced by 5-Fu and observed the dynamic changes of Oct3/4 by indirect immunofluorescence, Western blot, and quantitative real-time PCR. Immunofluorescence double staining was used to compare the positions of Oct3/4(+) cells and other reported alveolar epithelial stem cells. RESULTS: Oct3/4(+) cells were not found in normal rat lung epithelial cells. However, after treatment with 5-Fu, Oct3/4(+) cells appeared at 12 h, reached the peak at 24 h, then decreased at 48 h, and eventually disappeared at 72 h. Oct3/4 was localized in the nucleus. We found that the sites of Clara cell secretory protein and surfactant protein-C dual positive cells were apparently different from Oct3/4(+) cells. CONCLUSIONS: Our results revealed that, in rat alveolar epithelium, expression of Oct3/4 could be induced after treatment with 5-Fu, then decreased gradually, and was silenced following the alveolar epithelial differentiation. We hold that Oct3/4(+) cells are lung stem cells, which can provide new evidence for identification and isolation of lung epithelial stem cells.
Asunto(s)
Células Epiteliales Alveolares/metabolismo , Fluorouracilo/efectos adversos , Regulación de la Expresión Génica/efectos de los fármacos , Lesión Pulmonar/metabolismo , Factor 3 de Transcripción de Unión a Octámeros/biosíntesis , Mucosa Respiratoria/metabolismo , Células Madre/metabolismo , Células Epiteliales Alveolares/patología , Animales , Femenino , Fluorouracilo/farmacología , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/patología , Masculino , Ratas , Ratas Wistar , Reacción en Cadena en Tiempo Real de la Polimerasa , Mucosa Respiratoria/patología , Células Madre/patologíaRESUMEN
Accurate markers and molecular mechanisms of stem cell dormancy and activation are poorly understood. In this study, the anti-cancer drug, 5-fluorouracil, was used to selectively kill proliferating cells of human bronchial epithelial (HBE) cell line. This method can enrich and purify stem cell population. The dormant versus active status of stem cells was determined by phosphorylation of RNAp II Ser2. The surviving stem cells were cultured to form stem cell spheres expressing stem cell markers and transplanted into nude mice to form a teratoma. The results demonstrated the properties of stem cells and potential for multi-directional differentiation. Bisulfite sequencing polymerase chain reaction showed that demethylation of the Sox2 promoter by 5-FU resulted in Sox2 expression in the dormant stem cells. This study shows that the dormancy and activation of HBE stem cells is closely related to epigenetic modification.
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Bronquios/patología , Fluorouracilo/farmacología , Regulación Neoplásica de la Expresión Génica , Factores de Transcripción SOXB1/genética , Células Madre/patología , Teratoma/patología , Neoplasias de la Tráquea/patología , Animales , Antimetabolitos Antineoplásicos/farmacología , Apoptosis , Bronquios/efectos de los fármacos , Bronquios/metabolismo , Diferenciación Celular , Proliferación Celular , Metilación de ADN , Epigénesis Genética , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Regiones Promotoras Genéticas , ARN Polimerasa II/metabolismo , Factores de Transcripción SOXB1/metabolismo , Células Madre/efectos de los fármacos , Células Madre/metabolismo , Teratoma/tratamiento farmacológico , Teratoma/metabolismo , Neoplasias de la Tráquea/tratamiento farmacológico , Neoplasias de la Tráquea/metabolismo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Although the research on the localization of trachea stem cells has made a rapid progress, the mechanism of proliferation and differentiation of trachea stem cells remains unclear. The objective of this study is to observe and analyze the recovery process of mice tracheal epithelium injured by 5-FU, and to investigate the mechanism involved in the regulation of tracheal stem cells proliferation and differentiation through morphological, immunofluorescence, and microarray analysis. After treatment with 5-FU, the mature cells were dead and desquamated. Only a few G0 phase cells remained on the basement membrane. When supplied with normal culture media, the cells eventually became flat, cubic, and restored as pseudostratified epithelium. These G0 phase cells were ABCG2 positive. It suggested that these cells could differentiate into cilia cells or Clara cells, and had the multi-differentiation ability of stem cells. We examinated the expression profile of genes involved in the stem cell differentiation in normal tracheal epithelial cells and the regenerated epithelial cells at 24 and 48 h after injured by 5-FU using gene microarray. After 24 h treatment, 8 genes were up-regulated and 31 genes were down-regulated. After 48 h treatment, 5 genes were up-regulated and 42 genes were down-regulated. The differential gene expressions in gene microarray analysis focused on cell cycle regulation, intercellular junction, fibroblast growth factors, bone morphogenetic protein, Notch and Wnt-signaling pathways, which suggested that the differential gene expressions might be closely associated with the proliferation and differentiation of tracheal stem cells.
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Células Epiteliales/efectos de los fármacos , Fluorouracilo/toxicidad , Tráquea/citología , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2 , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Animales , Células Cultivadas , Regulación hacia Abajo/efectos de los fármacos , Células Epiteliales/citología , Células Epiteliales/metabolismo , Técnica del Anticuerpo Fluorescente , Ratones , Análisis de Secuencia por Matrices de Oligonucleótidos , Tráquea/patología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
BACKGROUND: Oct4 is a major transcription factor related to stem cell self-renewal and differentiation. To fulfill its functions, it must be able to enter the nucleus and remain there to affect transcription. KPNA2, a member of the karyopherin family, plays a central role in nucleocytoplasmic transport. The objective of the current study was to examine the association between Oct4 and KPNA2 expression levels with regard to both the clinicopathological characteristics and prognoses of patients with non-small-cell lung cancer (NSCLC). METHODS: Immunohistochemistry was used to detect the expression profile of Oct4 and KPNA2 in NSCLC tissues and adjacent noncancerous lung tissues. Real-time polymerase chain reaction and western blotting were used to detect the mRNA and protein expression profiles of Oct4 and KPNA2 in lung cancer cell lines. Small interfering RNAs were used to deplete Oct4 and KPNA2 expressions. Double immunofluorescence was used to detect Oct4 expression in KPNA2 knockdown cells. Co-immunoprecipitation was used to detect the interaction of Oct4 and KPNA2. RESULTS: Oct4 was overexpressed in 29 of 102 (28.4%) human lung cancer samples and correlated with differentiation (P = 0.002) and TNM stage (P = 0.003). KPNA2 was overexpressed in 56 of 102 (54.9%) human lung cancer samples and correlated with histology (P = 0.001) and differentiation (P = 0.045). Importantly, Oct4 and KPNA2 expression levels correlated significantly (P < 0.01). Expression of Oct4 and KPNA2 was associated with short overall survival. In addition, depleting Oct4 and KPNA2 expression using small interfering RNAs inhibited proliferation in lung cancer cell lines. Real-time polymerase chain reaction and western blotting analysis indicated that reduction of KPNA2 expression significantly reduced mRNA and nucleoprotein levels of Oct4. Double immunofluorescence analysis revealed that nuclear Oct4 signals were reduced significantly in KPNA2 knockdown cells. Co-immunoprecipitation experiments revealed that KPNA2 interacts with Oct4 in lung cancer cell lines. CONCLUSION: Oct4 and KPNA2 play an important role in NSCLC progression. Oct4 nuclear localization may be mediated by its interaction with KPNA2.
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Carcinoma de Pulmón de Células no Pequeñas/genética , Regulación hacia Abajo/genética , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Factor 3 de Transcripción de Unión a Octámeros/genética , alfa Carioferinas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular , Femenino , Humanos , Estimación de Kaplan-Meier , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Unión Proteica , alfa Carioferinas/metabolismoRESUMEN
Most adult stem cells are in the G0 or quiescent phase of the cell cycle and account for only a small percentage of the cells in the tissue. Thus, isolation of stem cells from tissues for further study represents a major challenge. This study sought to enrich cancer stem cells and explore cancer stem-like cell clones using 5-fluorouracil (5-FU) in the lung adenocarcinoma cell line, SPC. Proliferation inhibition was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, according to which half maximal inhibitory concentration values were calculated. Expression levels of stem cell markers after treatment with 5-FU were examined using immunofluorescence and Western blotting. Additionally, side population (SP) cells were sorted using FACS. Properties of SP cells were evaluated by using Transwell, colony-forming assays, and tumor formation experiments. 5-FU greatly inhibits proliferation, especially of cells in S phase. SP cells possess greater invasive potential, higher clone-forming potential, and greater tumor-forming ability than non-SP cells. Treatment with 5-FU enriches the SP cells with stem cell properties in human lung adenocarcinoma cell lines.
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Adenocarcinoma/tratamiento farmacológico , Antimetabolitos Antineoplásicos/farmacología , Biomarcadores de Tumor/metabolismo , Movimiento Celular/efectos de los fármacos , Fluorouracilo/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Células Madre Neoplásicas/efectos de los fármacos , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ensayo de Unidades Formadoras de Colonias , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos BALB C , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Células de Población Lateral/efectos de los fármacos , Células de Población Lateral/metabolismo , Células de Población Lateral/patología , Células Tumorales CultivadasRESUMEN
The present study examines clinicopathologic findings and their association with the Epstein-Barr virus (EBV) in Waldeyer's ring lymphomas (WRLs) from Indonesia (91 cases), P.R. China (31 cases), Korea (101 cases) and Japan (61 cases). Waldeyer's ring (WR) was categorized into upper and lower parts comprising the pharyngeal and tubal tonsils (upper WR) or palatine and lingual tonsils (lower WR), respectively. Diffuse large B-cell lymphoma (DLBCL) pre-dominated in the lower WR in all countries at a frequency of 78.9-100%. Natural killer/T-cell lymphoma (NKTCL) was predominant in the upper WR in China, Korea and Japan at a frequency of 50-62.5%, while in Indonesia it occcurred at a frequency of less than 10%. On the whole, patients with NKTCL were significantly younger (median 43 years) than those with DLBCL (57 years). Patients with DLBCL in the lower WR were significantly younger in Indonesia (median 50 years) than in China (63 years) or Japan (69 years). The percentage of EBV-positive cases was much higher in NKTCL (78.6-100%) than in DLBCL (2.2-6%). This study evaluates the differences between East and Southeast Asian countries in terms of histologic type and age distribution in WRLs categorized by the location of the lesions in WR.
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OBJECTIVE: To study the clinical features, immunophenotypes and the significance of Epstein-Barr virus infection in primary nasal and pharyngeal non-Hodgkin's lymphomas in Shenyang. METHODS: One hundred and fifty eight cases of primary nasal and pharyngeal non-Hodgkin's lymphomas were included in this study. The samples were stained with haematoxylin and eosin for histological examination. Immunohistochemistry studies were performed using monoclonal antibodies, including CD3 for T-lymphocytes, CD20 for B-lymphocytes, and CD56 and CD57 for NK cells. All cases were reclassified according to the new WHO classification of lymphomas (2001). In situ hybridization detection of EBV-encoded small nuclear RNA (EBER-1) was performed in 99 cases. RESULTS: Overall, 101 (63.9%) of the 158 NHL were extranodal NK/T cell lymphomas (nasal type), 23 (14.6%) were nonspecific peripheral T cell lymphomas and the remaining 34 cases (21.5%) were B cell lymphomas. The primary sites of involvement were the nasal cavity (53.2%, 84/158), the tonsil (24.7%, 39/158) and the pharynx (22.1%, 35/158). Among 99 cases studied by EBER-1 in situ hybridization, a positive detection was seen in 70/71 cases (98.6%) of extranodal NK/T cell lymphoma (nasal type), 8/12 cases (66.7%) of T cell lymphoma, and 7/16 cases (43.8%) of B cell lymphoma. CONCLUSIONS: Among primary nasal and pharyngeal NK lymphomas, extranodal NK/T cell lymphoma (nasal type) is the most common type and is strongly associated with EBV infection. The pathological diagnosis of nasal and pharyngeal lymphomas should take considerations of the anatomic sites and immunophenotypical features.
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Herpesvirus Humano 4/aislamiento & purificación , Linfoma no Hodgkin , Cavidad Nasal , Neoplasias Nasales , Neoplasias Faríngeas , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Complejo CD3/metabolismo , Antígeno CD56/metabolismo , Niño , Femenino , Humanos , Linfoma de Células B/metabolismo , Linfoma de Células B/patología , Linfoma de Células B/virología , Linfoma Extranodal de Células NK-T/metabolismo , Linfoma Extranodal de Células NK-T/patología , Linfoma Extranodal de Células NK-T/virología , Linfoma no Hodgkin/metabolismo , Linfoma no Hodgkin/patología , Linfoma no Hodgkin/virología , Linfoma de Células T Periférico/metabolismo , Linfoma de Células T Periférico/patología , Linfoma de Células T Periférico/virología , Masculino , Persona de Mediana Edad , Neoplasias Nasales/metabolismo , Neoplasias Nasales/patología , Neoplasias Nasales/virología , Neoplasias Faríngeas/metabolismo , Neoplasias Faríngeas/patología , Neoplasias Faríngeas/virología , ARN Viral/metabolismo , Neoplasias Tonsilares/metabolismo , Neoplasias Tonsilares/patología , Neoplasias Tonsilares/virología , Adulto JovenRESUMEN
Cases of nasal NK/T-cell lymphoma (NKTCL) occur occasionally in Asian and Latin American countries but rarely in Western countries. The etiological role of life-style and environmental factors in nasal NKTCL was investigated. Five university hospitals in Japan and one each in Korea and China participated in this study; a total of 88 cases and 305 hospital controls were accrued during 2000-2005. The odds ratio (OR) of NKTCL obtained after adjustments of age, sex and country was 4.15 (95% confidence interval (CI), 1.74-9.87) for farmers, 2.81 (CI, 1.49-5.29) for producers of crops, 4.01 (CI, 1.99-8.09) for pesticide users, 11.65 (CI, 1.17-115.82) for residents near garbage burning plants, 2.95 (CI, 1.25-6.95) for former drinkers, and 0.49 (CI, 0.23-1.04) for current smokers. The ORs for crop producers, who minimized their exposure to pesticides by using gloves and glasses, and sprinkling downwind at the time of pesticide use, were 3.30 (95% CI, 1.28-8.54), 1.18 (95% CI, 0.11-12.13) and 2.20 (95% CI, 0.88-5.53), respectively, which were lower than those for producers who did not take these precautions. Exposure to pesticides and chemical solvents could be causative of NKTCL. Taken together, life-style and environmental factors might be risk factors for NKTCL.
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Contaminantes Ambientales/toxicidad , Células Asesinas Naturales , Estilo de Vida , Linfoma de Células T/epidemiología , Neoplasias Nasales/epidemiología , Plaguicidas/toxicidad , Adulto , Anciano , Estudios de Casos y Controles , Asia Oriental/epidemiología , Femenino , Humanos , Incidencia , Linfoma de Células T/prevención & control , Masculino , Persona de Mediana Edad , Neoplasias Nasales/prevención & controlRESUMEN
OBJECTIVE: Localization of tracheal stem cells in rat trachea. METHODS: Extracorporeal tracheal injury (Wistar rats) was induced by 5-FU. The process of regeneration was observed and analyzed by light microscopy, electron microscopy, and immunohistochemistry. RESULTS: Twelve hours after treatment with 5-FU, the tracheal epithelium shed and cells with naked nuclei were seen located sparsely on the basement membrane. Six hours after removal of 5-FU, the tracheal rings were covered with flattened epithelium. These cells were poorly differentiated under electron microscopy. Immunohistochemistry showed few proliferating cell nuclear antigen (PCNA)-negative cells sparsely scattered among PCNA-positive cells on the basement membrane. Nine hours later, electron microscopy found that these cells differentiated into mucous cells and ciliated cells. Forty-eight hours later, the tracheal rings were entirely covered by pseudostratified ciliated columnar epithelium. CONCLUSIONS: A small number of G(0) cells with naked nuclei are located sparsely on the basement membrane of the trachea. Tracheal epithelium regenerates by proliferation and differentiation of these cells. It is likely that some of these G(0) cells on the tracheal basement membrane represent tracheal stem cells.