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Completely synthetic cell cultivation materials for human pluripotent stem cells (hPSCs) are important for the future clinical use of hPSC-derived cells. Currently, cell culture materials conjugated with extracellular matrix (ECM)-derived peptides are being prepared using only one specific integrin-targeting peptide. We designed dual peptide-conjugated hydrogels, for which each peptide was selected from different ECM sites: the laminin ß4 chain and fibronectin or vitronectin, which can target α6ß1 and α2ß1 or αVß5. hPSCs cultured on dual peptide-conjugated hydrogels, especially on hydrogels conjugated with peptides obtained from the laminin ß4 chain and vitronectin with a low peptide concentration of 200 µg/mL, showed high proliferation ability over the long term and differentiated into cells originating from 3 germ layers in vivo as well as a specific lineage of cardiac cells. The design of grafting peptides was also important, for which a joint segment and positive amino acids were added into the designed peptide. Because of the designed peptides on the hydrogels, only 200 µg/mL peptide solution was sufficient for grafting on the hydrogels, and the hydrogels supported hPSC cultures long-term; in contrast, in previous studies, greater than 1000 µg/mL peptide solution was needed for the grafting of peptides on cell culture materials.
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Human pluripotent stem cells (hPSCs) have the ability to differentiate into cells derived from three germ layers and are an attractive cell source for cell therapy in regenerative medicine. However, hPSCs cannot be cultured on conventional tissue culture flasks but can be cultured on biomaterials with specific hPSC integrin interaction sites. We designed hydrogels conjugated with several designed peptides that had laminin-ß4 active sites, optimal elasticities and different zeta potentials. A higher expansion fold of hPSCs cultured on the hydrogels was found with the increasing zeta potential of the hydrogels conjugated with designed peptides, where positive amino acid (lysine) insertion into the peptides promoted higher zeta potentials of the hydrogels and higher expansion folds of hPSCs when cultured on the hydrogels using xeno-free protocols. The hPSCs cultured on hydrogels conjugated with the optimal peptides showed a higher expansion fold than those on recombinant vitronectin-coated plates, which are the gold standard of hPSC cultivation dishes. The hPSCs could differentiate into specific cell lineages, such as mesenchymal stem cells (MSCs) and MSC-derived osteoblasts, even after being cultivated on hydrogels conjugated with optimal peptides for long periods of time, such as 10 passages.
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Hidrogeles , Células Madre Pluripotentes , Humanos , Hidrogeles/química , Proliferación Celular , Células Madre Pluripotentes/metabolismo , Péptidos/farmacología , Péptidos/metabolismo , Diferenciación CelularRESUMEN
Stem cells serve as an ideal source of tissue regeneration therapy because of their high stemness properties and regenerative activities. Mesenchymal stem cells (MSCs) are considered an excellent source of stem cell therapy because MSCs can be easily obtained without ethical concern and can differentiate into most types of cells in the human body. We prepared cell culture materials combined with synthetic polymeric materials of poly-N-isopropylacrylamide-co-butyl acrylate (PN) and extracellular matrix proteins to investigate the effect of cell culture biomaterials on the differentiation of dental pulp stem cells (DPSCs) into neuronal cells. The DPSCs cultured on poly-L-ornithine (PLO)-coated (TPS-PLO) plates and PLO and PN-coated (TPS-PLO-PN) plates showed excellent neuronal marker (ßIII-tubulin and nestin) expression and the highest expansion rate among the culture plates investigated in this study. This result suggests that the TPS-PLO and TPS-PN-PLO plates maintained stable DPSCs proliferation and had good capabilities of differentiating into neuronal cells. TPS-PLO and TPS-PN-PLO plates may have high potentials as cell culture biomaterials for the differentiation of MSCs into several neural cells, such as cells in the central nervous system, retinal cells, retinal organoids and oligodendrocytes, which will expand the sources of cells for stem cell therapies in the future.
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The transplantation of human mesenchymal stem cells (hMSCs), such as bone marrow stem cells (BMSCs) and adipose-derived stem cells (ADSCs), has shown beneficial effects in protecting transplanted tissues and cells against graft-versus-host disease (GVHD). Human pluripotent stem cell (hPSC)-derived mesenchymal stem cells (MSCs) can also be used to generate hMSCs with stable characteristics without limitations. Therefore, we differentiated human induced pluripotent stem cells (hiPSCs, H-M5) and human embryonic stem cells (hESCs, H9) into hMSCs on dishes coated with different extracellular matrix (ECM) proteins to study the effect of cell culture biomaterials on hPSC differentiation into hMSCs. hPSC-derived MSCs cultured on Matrigel (MAT)-coated, collagen (COL)-coated and laminin-521 (LN-521)-coated tissue culture polystyrene (TCP) dishes showed excellent proliferation speed and reduced aging over 10 passages. High MSC surface marker (CD44, CD73, CD90 and CD105) expression was also observed on hPSC-derived MSCs cultured on MAT-coated, COL-coated and LN-521-coated TCP dishes as well as uncoated TCP dishes. Analysis of late osteogenic differentiation by evaluation of mineral deposition revealed that hPSC-derived MSCs cultured on fibronectin (FN)-coated and LN-521-coated TCP dishes showed high osteogenic differentiation. ECM proteins are effective as coating materials on cell culture biomaterials to regulate the proliferation and differentiation fate of hPSC-derived MSCs.
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Diferenciación Celular , Proteínas de la Matriz Extracelular , Células Madre Pluripotentes Inducidas , Células Madre Mesenquimatosas , Materiales Biocompatibles/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/citología , Células Madre Mesenquimatosas/citología , OsteogénesisRESUMEN
Human pluripotent stem cells (hPSCs) are typically cultivated on extracellular matrix (ECM) protein-coated dishes in xeno-free culture conditions. We supplemented mixed ECM proteins (laminin-511 and recombinant vitronectin, rVT) in culture medium for hPSC culture on conventional polystyrene dishes. Three hPSC cell lines were successfully cultivated on uncoated polystyrene dishes in medium supplemented with optimal conditions of laminin-511 and rVT. Excellent colony shape and colony size as well as high expansion fold of hPSCs were found under these conditions, whereas the colony size was small and poor expansion fold was found solely on L-511-coated dishes. A small portion of L-511 in the culture medium supported hPSC adhesion and prevented the adhesion from being too strong on the uncoated dishes, and rVT in the culture medium further supported adhesion of hPSCs on the dishes by maintaining their pluripotency. Having the optimal composition of L-511 and rVT in the culture medium was important for generating good hPSC colony shapes and sizes as well as a high expansion fold. After long-term culture of hPSCs on uncoated dishes supplemented with the mixed proteins, the hPSCs successfully showed pluripotent markers and could differentiate into a specific lineage of cells, cardiomyocytes, with high efficiency.
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Laminina/metabolismo , Células Madre Pluripotentes/metabolismo , Poliestirenos/química , Vitronectina/metabolismo , Técnicas de Cultivo de Célula , Diferenciación Celular , Células Cultivadas , Humanos , Tamaño de la Partícula , Proteínas Recombinantes/metabolismo , Propiedades de SuperficieRESUMEN
Cancer stem cells (CSCs) or cancer-initiating cells (CICs) are key factors for tumor generation and metastasis. We investigated a filtration method to enhance CSCs (CICs) from colon carcinoma HT-29 cells and primary colon carcinoma cells derived from patient colon tumors using poly(lactide-co-glycolic acid)/silk screen (PLGA/SK) filters. The colon carcinoma cell solutions were permeated via porous filters to obtain a permeation solution. Then, the cell cultivation media were permeated via the filters to obtain the recovered solution, where the colon carcinoma cells that adhered to the filters were washed off into the recovered solution. Subsequently, the filters were incubated in the culture media to obtain the migrated cells via the filters. Colon carcinoma HT-29 cells with high tumorigenicity, which might be CSCs (CICs), were enhanced in the cells in the recovered solution and in the migrated cells based on the CSC (CIC) marker expression, colony-forming unit assay, and carcinoembryonic antigen (CEA) production. Although primary colon carcinoma cells isolated from colon tumor tissues contained fibroblast-like cells, the primary colon carcinoma cells were purified from fibroblast-like cells by filtration through PLGA/SK filters, indicating that the filtration method is effective in purifying primary colon carcinoma cells.
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We developed poly(vinyl alcohol-co-itaconic acid) (PV) hydrogels grafted with laminin-derived peptides that had different joint segments and several specific designs, including dual chain motifs. PV hydrogels grafted with a peptide derived from laminin-ß4 (PMQKMRGDVFSP) containing a joint segment, dual chain motif and cationic amino acid insertion could attach human pluripotent stem (hPS) cells and promoted high expansion folds in long-term culture (over 10 passages) with low differentiation rates, whereas hPS cells attached poorly on PV hydrogels grafted with laminin-α5 peptides that had joint segments with and without a cationic amino acid or on PV hydrogels grafted with laminin-ß4 peptides containing the joint segment only. The inclusion of a cationic amino acid in the laminin-ß4 peptide was critical for hPS cell attachment on PV hydrogels, which contributed to the zeta potential shifting to higher values (3-4 mV enhancement). The novel peptide segment-grafted PV hydrogels developed in this study supported hPS cell proliferation, which induced better hPS cell expansion than recombinant vitronectin-coated dishes (gold standard of hPS cell culture dishes) in xeno-free culture conditions. After long-term culture on peptide-grafted hydrogels, hPS cells could be induced to differentiate into specific lineages of cells, such as cardiomyocytes, with high efficiency.
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Hidrogeles/química , Péptidos/química , Polímeros/química , Secuencia de Aminoácidos , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Humanos , Hidrogeles/farmacología , Laminina/química , Ratones , Ratones Endogámicos NOD , Ratones SCID , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/metabolismo , Alcohol Polivinílico/química , Succinatos/química , Propiedades de SuperficieRESUMEN
The authors would like to make a correction to their published paper [...].
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INTRODUCTION: Coxsackievirus/adenovirus receptors (CARs) and desmoglein-2 (DSG2) are similar molecules to adenovirus-based vectors in the cell membrane. They have been found to be associated with lung epithelial cell tumorigenesis and can be useful markers in predicting survival outcome in lung adenocarcinoma (LUAD). METHODS: A gene ontology enrichment analysis disclosed that DSG2 was highly correlated with CAR. Survival analysis was then performed on 262 samples from the Cancer Genome Atlas, forming "Stage 1A" or "Stage 1B". We therefore analyzed a tissue microarray (TMA) comprised of 108 lung samples and an immunohistochemical assay. Computer counting software was used to calculate the H-score of the immune intensity. Cox regression and Kaplan-Meier analyses were used to determine the prognostic value. RESULTS: CAR and DSG2 genes are highly co-expressed in early stage LUAD and associated with significantly poorer survival (p = 0.0046). TMA also showed that CAR/DSG2 expressions were altered in lung cancer tissue. CAR in the TMA was correlated with proliferation, apoptosis, and epithelial-mesenchymal transition (EMT), while DSG2 was associated with proliferation only. The Kaplan-Meier survival analysis revealed that CAR, DSG2, or a co-expression of CAR/DSG2 was associated with poorer overall survival. CONCLUSIONS: The co-expression of CAR/DSG2 predicted a worse overall survival in LUAD. CAR combined with DSG2 expression can predict prognosis.
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The impact of the new International Association for the Study of Lung Cancer pathology committee grading system for advanced lung adenocarcinoma (LADC) on survival is unclear, especially in Asian populations. In this study, we reviewed the prognostic outcomes of patients with late-stage disease according to the new grading system. We reviewed 136 LADC cases who underwent a small biopsy from 2007 to 2018. Tumors were classified according to the new grading system for LADC. Baseline characteristics (age, sex, smoking status, body mass index, and driver gene mutations) were analyzed. Kaplan-Meier and Cox regression analyses were used to determine correlations with the new grading system and prognosis. Patients with poorly differentiated adenocarcinoma were significantly correlated with a poor progression-free survival (PFS) (p = 0.013) but not overall survival (OS) (p = 0.154). Subgroup analysis showed that wild-type EGFR patients with poorly differentiated adenocarcinoma treated with chemotherapy had significantly worse PFS (p = 0.011). There was no significant difference in survival among the patients with epidermal growth factor receptor mutations who were treated with tyrosine kinase inhibitors. Patients aged >70 years and those with a BMI ≤ 25 kg/m2 and wild-type patients had significantly worse OS in both univariate (HR = 1.822, p = 0.006; HR = 2.250, p = 0.004; HR = 1.537, p = 0.046, respectively) and multivariate analyses (HR = 1.984, p = 0.002; HR = 2.383, p = 0.002; HR = 1.632, p = 0.028, respectively). Despite therapy, patients with poorly differentiated tumors still fared worse than those with better differentiated tumors. No differences were found among the EGFR mutations treated with TKI. Our findings highlight that the therapeutic regimen should be adjusted for EGFR Wild-type patients with poorly differentiated adenocarcinoma treated with chemotherapy to provide better outcomes.
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Cancer-initiating cells (CICs) or cancer stem cells (CSCs) are primarily responsible for tumor initiation, growth, and metastasis and represent a few percent of the total tumor cell population. We designed a membrane filtration protocol to enrich CICs (CSCs) from the LoVo colon cancer cell line via nylon mesh filter membranes with 11 and 20 µm pore sizes and poly(lactide-co-glycolic acid)/silk screen (PLGA/silk screen) porous membranes (pore sizes of 20-30 µm). The colon cancer cell solution was filtered through the membranes to obtain a permeate solution. Subsequently, the cell culture medium was filtered through the membranes to collect the recovery solution where the cells attached to the membranes were rinsed off into the recovery solution. Then, the membranes were cultivated in the cultivation medium to collect the migrated cells from the membranes. The cells migrated from any membrane had higher expression of the CSC surface markers CD44 and CD133, had higher colony formation levels, and produced more carcinoembryonic antigen (CEA) than the colon cancer cells cultivated on conventional tissue culture plates (control). We established a method to enrich the CICs (CSCs) of colon cancer cells from migrated cells through porous polymeric membranes by the membrane filtration protocol developed in this study.
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Separación Celular/métodos , Neoplasias del Colon/patología , Filtración/métodos , Membranas Artificiales , Células Madre Neoplásicas/citología , Antígeno AC133/análisis , Antígeno AC133/metabolismo , Antígeno Carcinoembrionario/análisis , Antígeno Carcinoembrionario/metabolismo , Línea Celular Tumoral , Separación Celular/instrumentación , Filtración/instrumentación , Humanos , Receptores de Hialuranos/análisis , Receptores de Hialuranos/metabolismo , Nylons/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Porosidad , Seda/químicaRESUMEN
Human adipose-derived stem cells (hASCs) cultured for 5 passages were filtered through nylon (NY) mesh filter membranes coated with and without extracellular matrix proteins to obtain the permeation solution. Subsequently, the culture media were filtered via the membranes to obtain the recovery solution. Then, the membranes were cultured in cell culture medium to obtain the migrated cells from the membranes. The hASCs in the permeation solution, through any type of NY mesh filter membrane having 11 and 20 µm pore sizes, had lower osteogenic differentiation ability than conventional hASCs cultured on tissue culture polystyrene (TCP) dishes for passage 5, whereas the hASCs purified by the membrane migration method through NY mesh filter membranes coated with recombinant vitronectin, which have 11 and 20 µm pore sizes, showed a higher proliferation speed as well as higher osteogenic differentiation potential than the conventional hASCs cultured on TCP dishes for passage 5. The membrane filtration and migration methods would be useful for cell sorting for specific cells, such as hASCs with high proliferation and high osteogenic differentiation ability, which do not need antibody binding or genetic modification of the cells for the specific isolation of the cells.
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Tejido Adiposo/citología , Nylons/química , Células Madre/citología , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Filtración , Humanos , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Human mesenchymal stem cells (hMSCs), such as human adipose-derived stem cells (hADSCs), present heterogeneous characteristics, including varying differentiation abilities and genotypes. hADSCs isolated under different conditions exhibit differences in stemness. We isolated hADSCs from human fat tissues via culture on different cell culture biomaterials including tissue culture polystyrene (TCPS) dishes and extracellular matrix protein (ECM)-coated dishes in medium supplemented with 5% or 10% serum-converted human platelet lysate (hPL) or 10% fetal bovine serum (FBS) as a control. Currently, it is not clear whether xeno-free hPL in the cell culture medium promotes the ability of hMSCs such as hADSCs to differentiate into several cell lineages compared to the xenomaterial FBS. We investigated whether a synchronized effect of ECM (Matrigel, fibronectin, and recombinant vitronectin) coatings on TCPS dishes for efficient hADSC differentiation could be observed when hADSCs were cultured in hPL medium. We found that Matrigel-coated dishes promoted hADSC differentiation into osteoblasts and suppressed differentiation into chondrocytes in 10% hPL medium. Recombinant vitronectin- and fibronectin-coated dishes greatly promoted hADSC differentiation into osteoblasts and chondrocytes in 5% and 10% hPL media. hPL promoted hADSC differentiation into osteoblasts and chondrocytes compared to FBS on the fibronectin-coated surface and recombinant vitronectin-coated surface.
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Adipocitos/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Células Madre Mesenquimatosas/metabolismo , Adipocitos/citología , Diferenciación Celular , Células Cultivadas , Proteínas de la Matriz Extracelular/química , Humanos , Células Madre Mesenquimatosas/citología , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
BACKGROUND: Previous animal studies have shown that certain respiratory oncoviruses can lead to tumorigenesis, especially influenza virus. However, no clinical studies other than animal studies have been conducted to test this hypothesis. OBJECTIVE: To investigate the association between influenza and the risk of lung cancer using the Taiwan Cancer Registry Database (TCRD) and Taiwan's National Health Insurance Research Database (NHIRD). METHODS: We identified a study cohort consisting of patients aged 40 years or above who were enrolled in the NHIRD between 1 January 2012 and 31 December 2014. Among them, we identified patients with lung cancer (cases) and their matched controls (matched by age, sex, and disease risk score (DRS) at a ratio of 1:10). Multivariate conditional logistic regression models were used to evaluate the association between exposure to influenza (timing and cumulative number) and risk of lung cancer. RESULTS: We identified 32,063 cases and 320,627 matched controls. Influenza was associated with a 1.09-fold increased risk of lung cancer (aOR 1.09, 95% CI 1.04-1.14, p<0.0001). The risk of lung cancer increased slightly with cumulative exposure to influenza (1-2 exposures: aOR 1.05, 95% CI 1.00-1.11; 3-4 exposures: aOR 1.12, 95% CI 1.00-1.25; 5+ exposures: aOR 1.25, 95% CI 1.13-1.39). CONCLUSION: Exposure to influenza was associated with an increased risk of lung cancer and the risk increased with cumulative exposure to influenza. However, the lack of valid information on smoking could lead to confounding, and future studies collecting patients' smoking histories are warranted to validate the association between influenza and lung cancer.
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Gripe Humana/complicaciones , Neoplasias Pulmonares/etiología , Fumar/efectos adversos , Adulto , Estudios de Casos y Controles , Estudios de Cohortes , Bases de Datos Factuales , Femenino , Humanos , Modelos Logísticos , Neoplasias Pulmonares/epidemiología , Masculino , Persona de Mediana Edad , Riesgo , Taiwán/epidemiologíaRESUMEN
BACKGROUND: Concurrent mutations of synchronous multiple primary non-small cell lung cancer (SMPNSCLC) is rare, and only a few cases have been reported. Herein, we present a case of early-stage SMPNSCLC with T790M and L858R mutations. CASE PRESENTATION: A 68-year-old male patient presented to the Thoracic Surgery Department due to a tumor in the right lower lung. The tumor was detected more than 5 years previously during a health examination; however, the patient ignored the problem because the clinician at that time stated that the lesion was highly likely to be benign. Chest computed topography (CT) was ordered and the images showed a well-defined tumor in the right lower lung and a faint nodular lesion over the left lower lung field. A CT-guided biopsy results showed the presence of atypical cells and positive staining of TTF-1 and CK7. Surgical intervention was performed. The right- and left-sided tumors disclosed micropapillary predominant adenocarcinoma and acinar-predominant adenocarcinoma, respectively. Both tumors were positive for TTF-1 but negative for ALK and p40. Real-time PCR analysis showed that the right-sided tumor had an epidermal growth factor receptor (EGFR) mutation presenting as point mutation T790M in exon 20, while the left-sided tumor had a point mutation L858R in exon 21 of EGFR. CONCLUSIONS: Our patient's case suggests that tumors resembling a benign pattern with central calcification may be misdiagnosed. Thus, early screening for lung cancer is important, and intensive efforts to make a diagnosis through surgical resection or biopsies to allow for tailored optimal treatment may be preferential for the best patient outcomes.
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Adenocarcinoma del Pulmón/patología , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/patología , Mutación , Neoplasias Primarias Múltiples/patología , Adenocarcinoma del Pulmón/genética , Adenocarcinoma del Pulmón/cirugía , Anciano , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/cirugía , Receptores ErbB/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/cirugía , Masculino , Estadificación de Neoplasias , Neoplasias Primarias Múltiples/genética , Neoplasias Primarias Múltiples/cirugíaRESUMEN
Recombinant vitronectin-grafted hydrogels were developed by adjusting surface charge of the hydrogels with grafting of poly-l-lysine for optimal culture of human embryonic stem cells (hESCs) under xeno- and feeder-free culture conditions, with elasticity regulated by crosslinking time (10-30 kPa), in contrast to conventional recombinant vitronectin coating dishes, which have a fixed stiff surface (3 GPa). hESCs proliferated on the hydrogels for over 10 passages and differentiated into the cells derived from three germ layers indicating the maintenance of pluripotency. hESCs on the hydrogels differentiated into cardiomyocytes under xeno-free culture conditions with much higher efficiency (80% of cTnT+ cells) than those on conventional recombinant vitronectin or Matrigel-coating dishes just only after 12 days of induction. It is important to have an optimal design of cell culture biomaterials where biological cues (recombinant vitronectin) and physical cues (optimal elasticity) are combined for high differentiation of hESCs into specific cell lineages, such as cardiomyocytes, under xeno-free and feeder-free culture conditions.
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Células Madre Embrionarias Humanas/citología , Hidrogeles/química , Vitronectina/química , Diferenciación Celular/fisiología , Línea Celular , Proliferación Celular/fisiología , Células Madre Embrionarias Humanas/metabolismo , Humanos , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/metabolismoRESUMEN
PURPOSE: The purpose of this study was to evaluate content validity and feasibility of the Chinese language perineal assessment tool (PAT-C), to assess its use in the clinical setting, and establish an optimal cut point for identifying patients at high risk for incontinence-associated dermatitis (IAD). DESIGN: Psychometric evaluation of existing instrument. SUBJECTS AND SETTING: The sample comprised 440 patients managed in intensive care units of 3 hospitals across the island of Taiwan and an affiliated home care service. One hundred three nurses practicing throughout Taiwan participated in the workshops in the third phase of the experiment. METHODS: The content validity of the PAT-C was evaluated by 3 experienced nurses using the content validity index statistic. We calculated a receiver operating characteristics (ROC) curve to determine a cut point of high-risk IAD. The curve was based on assessment of patients from receiving care from the intensive care unit and home care service of Cathay General Hospital (located in Taipei, New Paipei and Hsinchu). Nurse perceptions on the feasibility of PAT-C were assessed using an investigator-developed survey. RESULTS: Three experienced nurses rated the PAT-C and gave a robust overall content validity index score of 97.22%. The cut point for identifying patients at high risk for developing IAD via ROC curve analysis of 440 patients was 7.5 (sensitivity: 0.85; specificity: 0.79, area under curve: 0.82, P value < .001). One hundred three enrolled nurses attended the workshops and evaluated the feasibility using the PAT-C. Most of the participants considered the PAT-C as necessary (97.90%), 49.7% of participants suggested IAD risk assessment should be implemented by first-line (generalist) nurses, and 40% of participants recommended assessment on a daily basis. CONCLUSIONS: Study findings indicate robust content validity, and results of the investigator survey of nurse perceptions of the PAT-C indicate the potential for its widespread use in the clinical setting. We found that a cut point score 8 or more indicates a high risk for developing IAD.
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Evaluación en Enfermería/normas , Perineo/lesiones , Enfermedades de la Piel/clasificación , Traducción , Anciano , Anciano de 80 o más Años , Área Bajo la Curva , Dermatitis/clasificación , Dermatitis/etiología , Incontinencia Fecal/complicaciones , Femenino , Humanos , Masculino , Persona de Mediana Edad , Evaluación en Enfermería/métodos , Curva ROC , Medición de Riesgo/métodos , Medición de Riesgo/normas , Enfermedades de la Piel/diagnóstico , Encuestas y Cuestionarios , Taiwán , Incontinencia Urinaria/complicacionesRESUMEN
BACKGROUND: Low-grade fibromyxoid sarcoma (LGFMS) is a rare soft tissue tumor that has a tendency to grow in the deep soft tissue of the trunk and extremities. Despite its benign appearance, the tumor has a high recurrence rate and metastatic potential. LGFMS in the perineal space is rare, and only a few cases have been reported. We present the first case of LGFMS to be located at the external anal sphincter. CASE PRESENTATION: A 27-year-old male patient admitted to our Surgical Department with perianal pain and swollen for a year. The digital rectal examination revealed a perianal mass. Oral metronidazole and analgesia were prescribed on suspicion of perianal abscess failed to alleviate the symptom; hence, the patient was scheduled for surgery. Intraoperative diagnosis revealed an encapsulated tumor in the external anal sphincter that extended from the perianal region orally to the pararectal space. The results of immunohistochemistry (MUC4 staining) and FUS gene rearrangement by fluorescence in situ hybridization confirmed the diagnosis of LGFMS. CONCLUSIONS: This case is unique in terms of the location of the rare soft tissue tumor. Although LGFMS is considered low grade, its unpredictable behavior necessitates a long-term follow-up.
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Canal Anal/patología , Fibroma/patología , Fibrosarcoma/patología , Adulto , Canal Anal/cirugía , Fibroma/cirugía , Fibrosarcoma/cirugía , Humanos , Masculino , Clasificación del Tumor , PronósticoRESUMEN
Human amniotic fluid-derived stem cells (hAFCs) are pluripotent fetal cells capable of differentiating into multiple lineages, including cell types of each of the three embryonic germ layers. Proper differentiation and maintenance of pluripotency, the defining characteristics of stem cells, are regulated not only by the cells themselves but also by their microenvironment. Furthermore, the physical characteristics of the cell culture materials, such as material elasticity, influence the results of stem cell differentiation. We investigated the osteogenic differentiation efficiency of hAFCs cultured on cell culture materials with different elasticities that were grafted with extracellular matrix-derived oligopeptides. Polyvinyl alcohol-co-itaconic acid (PV) hydrogels with different elasticities were prepared by controlling the crosslinking intensity, and the resulting PV hydrogels were grafted with and without extracellular matrix (ECM)-derived oligopeptides. Specific ECM-derived oligopeptides were used to maintain the pluripotency of AFCs and were determined by evaluation of pluripotent gene expression (Sox2 and Oct4). The osteogenic differentiation efficiency of the hAFCs, cultured on PV hydrogels grafted with and without ECM-derived oligopeptides, was analyzed by alkaline phosphatase activity, Alizarin Red S staining, and von Kossa staining. Unmodified PV hydrogels induced osteoblast differentiation of hAFCs with high efficiency. We conclude that the hAFCs interacting with ECM-derived oligopeptides tended to maintain an undifferentiated state.
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KRAS and BRAF mutations are frequently detected in cases of colorectal cancer (CRC). The microsatellite status of patients with CRC and mutated KRAS/BRAF is important when determining cancer therapy. In the present study, the microsatellite status and genetic polymorphisms of KRAS (codons 12 and 13) and BRAF (V600E) were characterized in CRC tissue. The mismatch repair activity and oncogenic potential of KRAS were assessed by immunoblots from two KRAS-mutated CRC cell lines, SW480 and HCT116, with different microsatellite statuses, following treatment with 5-fluorouracil (5-FU) and oxaliplatin. Of all the 205 patients with CRC enrolled in the present study, 31.2% (64 of 205) had a KRAS or BRAF mutation, and 79.7% (51 of 64) of these patients with a KRAS/BRAF mutation exhibited microsatellite stability (MSS), indicating that microsatellite status is correlated with KRAS/BRAF mutation (P=0.027). A higher proportion (39.0%, 41 of 105) of elderly patients (≥62.6 years) had mutated KRAS or BRAF than younger patients (<62.6 years; 23.0%, 23 of 100; P=0.013). In the subgroup of 154 patients with MSS, patients without the KRAS or BRAF mutation (n=110) had longer disease-specific survival rates (58.8±9.4%) than patients with KRAS or BRAF mutations (n=44; 50.6±11.0%; P=0.043). Cytoplasmic KRAS levels decreased whereas nuclear MutS protein homolog 2 (MSH2) levels increased slightly in CRC HCT116 cells that were microsatellite instable, following treatment with 76.9 µM 5-FU for 2 days. In microsatellite stable SW480 cells, MSH2 levels markedly increased in the nucleus following 150 µM oxaliplatin treatment for 3 days. However, no significant change was observed regarding KRAS distribution in these cells. The results of the present study suggest that it is important to identify patients with CRC who may benefit from adjuvant chemotherapy with 5-FU or oxaliplatin, particularly CRC patients with MSS and mutated KRAS or BRAF, who have poorer overall survival rates than patients with microsatellite instability. Knowledge of the microsatellite status of patients and whether they harbor KRAS or BRAF mutations may enable more effective therapeutic strategies to be developed. Further prospective studies are required to validate the findings of the current study.