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931.
Nagamoto Y Tashiro K Takayama K Ohashi K Kawabata K Sakurai F Tachibana M Hayakawa T Furue MK Mizuguchi H 《Biomaterials》2012,33(18):4526-4534
Hepatocyte-like cells differentiated from human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) are known to be a useful cell source for drug screening. We recently developed an efficient hepatic differentiation method from hESCs and hiPSCs by sequential transduction of FOXA2 and HNF1α. It is known that the combination of three-dimensional (3D) culture and co-culture, namely 3D co-culture, can maintain the functions of primary hepatocytes. However, hepatic maturation of hESC- or hiPSC-derived hepatocyte-like cells (hEHs or hiPHs, respectively) by 3D co-culture systems has not been examined. Therefore, we utilized a cell sheet engineering technology to promote hepatic maturation. The gene expression levels of hepatocyte-related markers (such as cytochrome P450 enzymes and conjugating enzymes) and the amount of albumin secretion in the hEHs or hiPHs, which were 3D co-cultured with the Swiss 3T3 cell sheet, were significantly up-regulated in comparison with those in the hEHs or hiPHs cultured in a monolayer. Furthermore, we found that type I collagen synthesized in Swiss 3T3 cells plays an important role in hepatic maturation. The hEHs or hiPHs that were 3D co-cultured with the Swiss 3T3 cell sheet would be powerful tools for medical applications, such as drug screening. 相似文献
932.
Shinya Sakai Hiroshi Ochiai Hiroshi Kawamata Toshiaki Kogure Yutaka Shimada Katsuhisa Nakajima Katsutoshi Terasawa 《Journal of medical virology》1997,53(2):145-149
The production of several inflammatory cytokines, such as murine macrophage inflammatory protein-2 (MIP-2), tumor necrosis factor (TNF), and interleukin (IL)-1, was investigated in response to respiratory syncytial virus (RSV) infection in a murine macrophage cell line, RAW264.7, with special reference to mutual relation of their productions. The kinetics of MIP-2 production showed a trend for a biphasic pattern, that is, MIP-2 levels became detectable from 2 h postinfection (p.i.) and increased markedly until 8 h p.i. Thereafter, this level fell to the same level until 16 h p.i. and then increased again. TNF α was also detectable at 2 h p.i. and then increased sharply until 8 h p.i., when the peak level attained. Compared with the levels of MIP-2 and TNF α, that of IL-1 α/β, especially IL-1 β, was lower (ng versus pg/ml order). The presence of anti-TNF α or anti-IL-1 α antibody did not influence the early phase of MIP-2 production but significantly inhibited the late phase, suggesting that MIP-2 is induced by the combined effects of RSV infection via direct induction and indirectly after initial induction of TNF α and IL-1 α productions. Although RSV-infected RAW264.7 cells had no alteration inviability compared with mock-infected control, these data demonstrate that RSV is a potent inducer of inflammatory cytokines by direct induction and indirectly via the initial production of other cytokines. J. Med. Virol. 53:145–149, 1997. © 1997 Wiley-Liss, Inc. 相似文献
933.
Nagano S Otsuka T Niiro H Yamaoka K Arinobu Y Ogami E Akahoshi M Inoue Y Miyake K Nakashima H Niho Y Harada M 《International immunology》2002,14(7):733-740
Neutrophils are an important cellular source of proinflammatory mediators, whose regulation may be of potential benefit for the treatment of a number of inflammatory diseases. However, the mechanisms of lipopolysaccharide (LPS)-induced neutrophil activation and its regulation by anti-inflammatory cytokines have not yet been fully elucidated. Recent studies have revealed that mitogen-activated protein kinases (MAPK) play a crucial role in the generation of proinflammatory mediators in some cell types. Therefore, we conducted this study to determine whether MAPK activation could be involved in prostaglandin E(2) (PGE(2)) production and cyclooxygenase (COX)-2 expression in LPS-stimulated human neutrophils. PD98059 (MEK1 inhibitor) and SB203580 (p38(MAPK) inhibitor) reduced PGE(2) production as well as COX-2 expression in LPS-stimulated neutrophils. In addition, both extracellular signal-regulated protein kinase (ERK) and p38(MAPK) were phosphorylated and activated in time- and dose-dependent manners. Since we previously showed that IL-10 and IL-4 similarly inhibited COX-2 expression in LPS-stimulated neutrophils, we next tested the effects of IL-10 and IL-4 on the phosphorylation and activation of both kinases. IL-10 inhibited the phosphorylation and activation of p38(MAPK), but not ERK. In addition, IL-4 caused a marginal inhibition in the activation of p38(MAPK). Taken together, these results suggest that both ERK and p38(MAPK) pathways are involved in LPS-induced COX-2 expression and PGE(2) production in neutrophils, and IL-10 and IL-4 inhibit neutrophil prostanoid synthesis by down-regulating the activation of p38(MAPK). 相似文献
934.
Utilization of healthcare resources in obstructive sleep apnea syndrome: a 5-year follow-up study in men using CPAP 总被引:3,自引:0,他引:3
STUDY OBJECTIVES: Patients with untreated obstructive sleep apnea syndrome (OSAS) have higher healthcare utilization than matched controls. However, the long-term impact of continuous positive airway pressure (CPAP) use on healthcare utilization is unknown. DESIGN: Retrospective observational cohort study. SUBJECTS: There were 342 eligible men with OSAS and matched controls on whom there were utilization data for 5 years prior to initial OSAS diagnosis and for the 5 years on CPAP treatment of the cases. INTERVENTIONS: Patients were treated with CPAP. RESULTS: Patients with OSAS were typical cases (mean +/- SD): age, 48.2 +/- 0.6 years; body mass index, 35.6 +/- 0.4 kg/m2; Epworth Sleepiness Scale score, 14.2 +/- 0.3; apnea-hypopnea index, 47.1 +/- 1.8 events per hour. The number of physician visits were higher by 3.46 +/- 0.2 (95% confidence interval [CI]: 2.57 to 4.36) in cases in the year before diagnosis, compared with the fifth year before diagnosis, then decreased over the next 5 years by 1.03 +/- 0.49 (95% CI: -1.99 to -0.07)(P<.0001). Physician fees, in Canadian dollars, were higher by dollars 148.65 +/- dollars 27.27 (95% CI: 95.12 to 202.10) in cases in the year before diagnosis, compared with the fifth year before diagnosis, and then decreased over the next 5 years by dollars 13.92 +/- dollars 27.94(95%CI: -68.68 to 40.83)(P=.0009). Preexisting ischemic heart disease at the time of OSAS diagnosis predicted about a 5-fold increase in healthcare utilization between the second and fifth year of treatment. CONCLUSIONS: Treatment of OSAS reversed the trend of increasing healthcare utilization seen prior to diagnosis. Preexisting ischemic heart disease results in a negative impact on healthcare utilization. CPAP results in a long-term health benefit, as measured by the use of healthcare services. 相似文献
935.
Sekine H Shimizu T Dobashi I Matsuura K Hagiwara N Takahashi M Kobayashi E Yamato M Okano T 《Tissue engineering. Part A》2011,17(23-24):2973-2980
Regenerative therapies have currently emerged as one of the most promising treatments for repair of the damaged heart. Recently, numerous researchers reported that isolated cell injection treatments can improve heart function in myocardial infarction models. However, significant cell loss due to primary hypoxia or cell wash-out and difficulty to control the location of the grafted cells remains problem. As an attempt to overcome these limitations, we have proposed cell sheet-based tissue engineering, which involves stacking confluently cultured cells (two-dimensional), cell sheets, to construct three-dimensional cell-dense tissues. Cell sheet transplantation has been able to recover damaged heart function. However, no detailed analysis for transplanted cell survival has been previously performed. The present study compared the survival of cardiac cell sheet transplantation to direct cell injection in a rat myocardial infarction model. Luciferase-expressing neonatal rat cardiac cells were harvested as cell sheets from temperature-responsive culture dishes. The transplantation of cell sheets was compared to the direct injection of isolated cells dissociated with trypsin-ethylenediaminetetraacetic acid. These grafts were transplanted to infarcted rat hearts and cardiac function was assessed by echocardiography at 2 and 4 weeks after transplantation. In vivo bioluminescence and histological analyses were performed to examine cell survival. Cell sheet transplantation consistently yielded greater cell survival than cell injection. Immunohistochemistry revealed that cardiac cell sheets existed over the infarcted area as an intact layer. In contrast, the injected cells were scattered with relatively few cardiomyocytes in the infarcted areas. Four weeks after transplantation, cardiac function was also significantly improved in the cell sheet transplantation group compared with the cell injection. Twenty-four hours after cell grafting, significantly greater numbers of mature capillaries were also observed in the cardiac cell sheet transplantation. Additionally, the numbers of apoptotic cells with deterioration of integrin-mediated attachment were significantly lower after cardiac cell sheet transplantation. In accordance with increased cell survival, cardiac function was significantly improved after cardiac cell sheet transplantation in comparison to cell injection. Cell sheet transplantation can repair damaged hearts through improved cell survival and should become a promising therapy in cardiovascular regenerative medicine. 相似文献
936.