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Hsu C.-C.,Chang Gung Memorial Hospital at Keelung | Hsu C.-C.,Chang Gung University | Tsai W.-C.,Chang Gung University | Chen C.P.-C.,Chang Gung Memorial Hospital at Linkou | And 2 more authors.
American Journal of Physiology - Cell Physiology | Year: 2010

Negative-pressure wound therapy has recently gained popularity in chronic wound care. This study attempted to explore effects of different negative pressures on epithelial migration in the woundhealing process. The electric cell-substrate impedance sensing (ECIS) technique was used to create a 5 × 10-4 cm2 wound in the Madin-Darby canine kidney (MDCK) and human keratinocyte (HaCaT) cells. The wounded cells were cultured in a negative pressure incubator at ambient pressure (AP) and negative pressures of 75 mmHg (NP75), 125 mmHg (NP125), and 175 mmHg (NP 175). The effective time (ET), complete wound healing time (T max), healing rate (Rheal), cell diameter, and wound area over time at different pressures were evaluated. Traditional wound-healing assays were prepared for fluorescent staining of cells viability, cell junction proteins, including ZO-1 and E-cadherin, and actins. Amount of cell junction proteins at AP and NP125 was also quantified. In MDCK cells, the ET (1.25 ± 0.27 h), Tmax (1.76 ±0.32 h), and R heal (2.94 ± 0.62 × 10-4 cm2/h) at NP125 were significantly (P < 0.01) different from those at three other pressure conditions. In HaCaT cells, the Tmax (7.34 ± 0.29 h) and Rheal (6.82 ± 0.26 × 10-5 cm2/h) at NP125 were significantly (P < 0.01) different from those at NP75. Prominent cell migration features were identified in cells at the specific negative pressure. Cell migration activities at different pressures can be documented with the real-time wound-healing measurement system. Negative pressure of 125 mmHg can help disassemble the cell junction to enhance epithelial migration and subsequently result in quick wound closure. Copyright © 2010 the American Physiological Society.

Wu C.-T.,Chang Gung Memorial Hospital at Keelung | Wu C.-T.,Chang Gung University | Lin W.-Y.,Chang Gung University | Chang Y.-H.,Chang Gung University | And 3 more authors.
Oncotarget | Year: 2015

The aim of this study was to examine the role of miRNAs regulation by DNMT1 and its underlying mechanisms in bladder cancer. The choice of target miRNAs was based on the analysis of a TaqMan MicroRNA Panel assay. The role of target miRNA in tumor behavior and the related signaling pathways were assessed using the human bladder cancer cell lines. We also evaluated the predictive power of the target miRNA and its link to DNMT1 from 124 clinical bladder cancer specimens. Our results revealed that the miR-424 level is significantly increased when blocking DNMT1 in bladder cancer cells. From the clinical specimen analysis, the staining of miR-424 was inversely correlated with DNMT1 immunoreactivity. The lack of miR-424 expression was significantly linked to aggressive tumor growth, advanced clinical stage and poor prognosis in bladder cancer. Increased miR-424 suppressed the tumor growth rate and invasion ability determined in vitro and in vivo. Furthermore, the EGFR pathway plays a role in the transmission of the miR-424 signal that regulates cell growth and the epithelial-to-mesenchymal transition. These results highlight a potential role for miR-424 as a molecular predictor and therapeutic target in bladder cancer.

Wu H.-P.,Chang Gung University | Kuo S.-F.,Chang Gung University | Wu S.-Y.,Chang Gung Memorial Hospital at Keelung | Chuang D.-Y.,National Chung Hsing University
Cytokine | Year: 2010

Sugar control is important in patients with sepsis. Interleukin (IL)-12 induces the polarization of CD4+ T cells to the T helper 1 (Th1) phenotype. Regulatory T (Treg) cells are important in immunity and disease. The aim of this work is to determine whether hyperglycemia or insulin alters IL-12 response in peripheral mononuclear cells (PBMCs). Methods: The PBMCs from 15 type 2 diabetes mellitus (DM) patients and 13 healthy controls were used for cell analysis and culture with or without treatment by glucose and insulin or stimulation by lipopolysaccharide (LPS) for 1, 2, and 3. days. Results: The IL-12 level in the supernatant of LPS-stimulated PBMCs in the DM patients was significantly higher than that of healthy controls from day 1 to day 3. Kinetic IL-12 responses of LPS-stimulated PBMCs in the DM patients from day 1 to day 3 were significantly higher than that in healthy controls. The LPS-stimulated PBMCs under glucose treatment produced more IL-12 in DM patients but this did not happen in healthy controls. In DM patients, insulin could suppress IL-12 production from stimulated PBMCs but not with additional glucose treatment. Conclusion: The PBMCs of LPS-treated DM patients produced more IL-12 than that of LPS-treated healthy controls did. Hyperglycemia influenced IL-12 response from PBMCs in DM patients to some degree during infection. © 2010 Elsevier Ltd.

Chen H.-C.,Chang Gung Memorial Hospital at Keelung | Chen H.-C.,Chang Gung University | Chen H.-C.,Taipei Medical University | Fong T.-H.,Taipei Medical University | And 2 more authors.
Journal of Surgical Research | Year: 2013

Background: Spinal cord injuries (SCIs) are serious and debilitating health problems that lead to severe and permanent neurological deficits resulting from the primary mechanical impact followed by secondary tissue injury. During the acute stage after an SCI, the expression of autophagy and inflammatory responses contribute to the development of secondary injury. In the present study, we examined the multifaceted effects of rapamycin on outcomes of rats after an SCI. Materials and methods: We used 72 female Sprague-Dawley rats for this study. In the SCI group, we performed a laminectomy at T10, followed by impact-contusion of the spinal cord. In the control group, we performed only a laminectomy without contusion. We evaluated the effects of rapamycin using the Basso, Beattie, and Bresnahan scale for functional outcomes, Western blot analyses for analyzing LC3-II, tumor necrosis factor expression, and p70S6K phosphorylation, and an immunostaining technique for localization and enumeration of microglial and neuronal cells. Results: Basso, Beattie, and Bresnahan scores after injury significantly improved in the rapamycin-treated group compared with the vehicle group (on Day 28 after the SCI; P <.05). The Western blot analysis demonstrated that rapamycin enhanced LC3-II expression and decreased p70S6K phosphorylation compared with the vehicle (P <.01), which implies promotion of autophagy through mammalian target of rapamycin inhibition. Furthermore, rapamycin treatment significantly attenuated tumor necrosis factor production and microglial expression (P <.05). Immunohistochemistry of NeuN (antibodies specific to neurons) showed remarkable neuronal cell preservation in the rapamycin-treated group compared with the vehicle-treated group (P <.05), which suggests a neuroprotective effect of rapamycin. Conclusions: Rapamycin is a novel neuroprotectant with multifaceted effects on the rat spinal cord after injury. Use of such a clinically established drug could facilitate early clinical trials in selected cases of human SCIs. © 2013 Elsevier Inc. All rights reserved.

Wang L.-J.,Chang Gung University | Chen C.-K.,Chang Gung Memorial Hospital at Keelung | Chen C.-K.,Chang Gung University | Huang Y.-S.,Chang Gung University | Huang Y.-S.,Sleep Center
Journal of Child and Adolescent Psychopharmacology | Year: 2015

Objective: This study investigated the trends in neurocognitive function and behavioral symptoms among patients with attention-deficit/hyperactivity disorder (ADHD) during 24 months of treatment with methylphenidate in a clinical setting. Methods: Study participants consisted of 181 ADHD patients with a mean age of 13.4±2.5 years (ages ranged from 8 to 18 years; 151 boys and 30 girls) who were prescribed oral short-acting methylphenidate two or three times daily, with each dose ranging between 0.3 and 1.0mg/kg. At baseline and 6, 12, 18, and 24 months from baseline, neurocognitive function was assessed using the Test of Variables of Attention (TOVA) on the day the patient was off medication, and behavioral symptoms were evaluated using the Swanson, Nolan, and Pelham Version IV Scale for ADHD (SNAP-IV) parent form, the SNAP-IV teacher form, and the ADHD-Rating Scale (completed by a child psychiatrist). Results: Of the 181 ADHD patients at the initial visit, 103 (56.9%) completed the study. During the 24-month methylphenidate treatment, only the commission errors in TOVA significantly improved; however, the omission errors, response time, response time variability, response sensitivity, and ADHD score did not. The behavioral symptoms of ADHD, observed by various informants, all declined substantially, and were significantly correlated with each other during the long-term follow-up. The severity of teacher ratings was lower than that of parent and psychiatrist ratings. However, the teacher-rated inattention symptoms showed the strongest correlations with TOVA performance. Conclusions: Findings suggest that neurocognitive deficits in ADHD patients, except inhibition ability, might be long lasting in realistic settings. In addition, obtaining behavior profile assessments from multiple informants, especially from teachers, is vital for establishing a complete understanding of ADHD patients. © Copyright 2015, Mary Ann Liebert, Inc.

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