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Objective: To evaluate the therapeutic efficacy of hematopoietic stem cell transplantation (HSCT) for Wiskott-Aldrich syndrome (WAS), and to analyze the factors related to the outcomes. Methods: The clinical data of 60 children with WAS received HSCT in Shanghai Children's Medical Center from January 2006 to December 2020 were retrospectively analyzed. All cases were treated with a myeloablative conditioning regimen with busulfan and cyclophosphamide, and a graft-versus-host disease (GVHD) prevention regimen based on cyclosporine and methotrexate. Implantation, GVHD, transplant-related complications, immune reconstitution and survival rate were observed. Survival analysis was performed by Kaplan-Meier method, and Log-Rank method was used for univariate comparison. Results: The 60 male patients had main clinical features as infection and bleeding. The age at diagnosis was 0.4 (0.3, 0.8) years, and the age at transplantation was 1.1 (0.6, 2.1) years. There were 20 cases of human leukocyte antigen matched transplantation and 40 mismatched transplantation; 35 patients received peripheral blood HSCT, and 25 cord blood HSCT. All cases were fully implanted. The incidence of acute GVHD (aGVHD) was 48% (29/60) and only 2 (7%) developed aGVHD of grade â ¢; the incidence of chronic GVHD (cGVHD) was 23% (13/56), and all cases were limited. The incidence of CMV and EBV infection was 35% (21/60) and 33% (20/60) respectively; and 7 patients developed CMV retinitis. The incidence of sinus obstruction syndrome was 8% (5/60), of whom 2 patients died. There were 7 cases (12%) of autoimmune hemocytopenia after transplantation. Natural killer cells were the earliest to recover after transplantation, and B cells and CD4+T cells returned to normal at about 180 days post HSCT. The 5-year overall survival rate (OS) of this group was 93% (95%CI 86%-99%), and the event free survial rate (EFS) was 87% (95%CI 78%-95%). EFS of non-CMV reactivation group is higher than that of CMV reactivation group (95% (37/39) vs.71% (15/21), χ2=5.22, P=0.022). Conclusions: The therapeutic efficacy of HSCT for WAS is satisfying, and the early application of HSCT in typical cases can achieve better outcome. CMV infection is the main factor affecting disease-free survival rate, which can be improved by strengthening the management of complications.
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Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Síndrome de Wiskott-Aldrich , Humanos , Masculino , Niño , Estudios Retrospectivos , Síndrome de Wiskott-Aldrich/terapia , China , Trasplante de Células Madre Hematopoyéticas/métodos , Enfermedad Injerto contra Huésped/prevención & control , Acondicionamiento PretrasplanteRESUMEN
Coaxial electrospray is advantageous for the production of multidrug-releasing nanocarriers because it permits precise control over particle size, inhibits initial burst release, and offers moderate preparation conditions. In this study, a single-step coaxial electrospray technique is presented that achieves over 90 % co-encapsulation of the saxagliptin and dapagliflozin, two drugs treating type 2 diabetes, into biodegradable poly (d,l-lactide-co-glicolide) (PLGA) nanoparticles. Scanning electron microscopy reveals spherical and smooth shapes with diameters ranging from 534.8 to 708.6 nm. Transmission electron microscopy revealed clear core-shell and trilayer nanostructures. Fourier transform infrared spectroscopy confirmed the presence of PLGA, saxagliptin, and dapagliflozin in all the evaluated formulations. The results of the drug release investigation indicated the prolonged and regulated release of saxagliptin and dapagliflozin from bi- and trilayer structures, as compared to monolayer particles. Computational modelling showed good agreement with the experimental drug release profile in vitro. Further, cytotoxicity assay demonstrates that the formulated nanoparticles display good cytocompatibility. This study indicates that with the controllable and distinctive sustained release profiles, the hybrid nanoparticle-based drug delivery system can effectively co-encapsulate multiple drugs treating type 2 diabetes in a protectively shell of PLGA for therapeutically-benefit controlled release.
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Diabetes Mellitus Tipo 2 , Nanopartículas , Humanos , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Nanopartículas/química , Sistemas de Liberación de Medicamentos , Tamaño de la Partícula , Portadores de FármacosRESUMEN
Long-term stability of microbubbles is crucial to their effectiveness. Using a new microfluidic device connecting three T-junction channels of 100 µm in series, stable monodisperse SiQD-loaded bovine serum albumin (BSA) protein microbubbles down to 22.8 ± 1.4 µm in diameter were generated. Fluorescence microscopy confirmed the integration of SiQD on the microbubble surface, which retained the same morphology as those without SiQD. The microbubble diameter and stability in air were manipulated through appropriate selection of T-junction numbers, capillary diameter, liquid flow rate, and BSA and SiQD concentrations. A predictive computational model was developed from the experimental data, and the number of T-junctions was incorporated into this model as one of the variables. It was illustrated that the diameter of the monodisperse microbubbles generated can be tailored by combining up to three T-junctions in series, while the operating parameters were kept constant. Computational modeling of microbubble diameter and stability agreed with experimental data. The lifetime of microbubbles increased with increasing T-junction number and higher concentrations of BSA and SiQD. The present research sheds light on a potential new route employing SiQD and triple T-junctions to form stable, monodisperse, multi-layered, and well-characterized protein and quantum dot-loaded protein microbubbles with enhanced stability for the first time.
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Microburbujas , Puntos Cuánticos , Dispositivos Laboratorio en un Chip , Microfluídica , Albúmina Sérica Bovina , SilicioRESUMEN
OBJECTIVE: Colorectal cancer (CRC) has a very high morbidity and mortality worldwide. Related studies have shown that microRNA-543 (miR-543) is involved in the development of many cancers, including CRC. The purpose of this study was to explore the potential molecular mechanism of miR-543's involvement in the development of CRC. PATIENTS AND METHODS: QRT-PCR and Western blot were used to detect the expression of proliferation and migration-related proteins, signal transduction and transcriptional activator 3 and protein inhibitor of activated signal transducer and activators of transcription 3 (PIAS3). Cell proliferation and metastasis were measured by MTT, transwell and Western blot. The binding sites of miR-543 and PIAS3 were predicted by TargetScan database and verified by double-luciferase report experiment. RESULTS: The expression of miR-543 was high in CRC tissues and cell lines, while the mRNA and protein levels of PIAS3 were decreased. Meanwhile, a negative correlation between miR-543 and PIAS3 was also observed in CRC tissues. Moreover, the downregulation of miR-543 led to the inhibition of viability and the expression of proliferation and migration related proteins. Subsequently, miR-543 depletion also blocked cell migration and invasion. MiR-543 inhibits the expression of PISA3. Furthermore, downregulation of PIAS3 undermined the miR-543 depletion-mediated suppression effect on SW480 and LOVO cells. Notably, loss of miR-543 downregulated STAT3 activity, which was rescued by PIAS3 ablation. CONCLUSIONS: MiR-543 participated in cell proliferation and metastasis by targeting PIAS3 in CRC.
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Neoplasias Colorrectales , MicroARNs , Chaperonas Moleculares , Proteínas Inhibidoras de STAT Activados , Línea Celular , Movimiento Celular/genética , Proliferación Celular/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Células Epiteliales/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Proteínas Inhibidoras de STAT Activados/genética , Proteínas Inhibidoras de STAT Activados/metabolismoRESUMEN
HYPOTHESIS: Developing oral formulations to enable effective release of poorly water-soluble drugs like progesterone is a major challenge in pharmaceutics. Coaxial electrospray can generate drug-loaded nanoparticles of strategic compositions and configurations to enhance physiological dissolution and bioavailability of poorly water-soluble drug progesterone. EXPERIMENTS: Six formulations comprising nanoparticles encapsulating progesterone in different poly(lactide-co-glycolide) (PLGA) matrix configurations and compositions were fabricated and characterized in terms of morphology, molecular crystallinity, drug encapsulation efficiency and release behavior. FINDINGS: A protocol of fabrication conditions to achieve 100% drug encapsulation efficiency in nanoparticles was developed. Scanning electron microscopy shows smooth and spherical morphology of 472.1±54.8 to 588.0±92.1 nm in diameter. Multiphoton Airyscan super-resolution confocal microscopy revealed core-shell nanoparticle configuration. Fourier transform infrared spectroscopy confirmed presence of PLGA and progesterone in all formulations. Diffractometry indicated amorphous state of the encapsulated drug. UV-vis spectroscopy showed drug release increased with hydrophilic copolymer glycolide ratio while core-shell formulations with progesterone co-dissolved in PLGA core exhibited enhanced release over five hours at 79.9±1.4% and 70.7±3.5% for LA:GA 50:50 and 75:25 in comparison with pure progesterone without polymer matrix in the core at 67.0±1.7% and 57.5±2.8%, respectively. Computational modeling showed good agreement with the experimental drug release behavior in vitro.
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Nanopartículas/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Progesterona/administración & dosificación , Progesterona/farmacocinética , Disponibilidad Biológica , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Composición de Medicamentos/métodos , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Microscopía Electrónica de Rastreo , Nanopartículas/administración & dosificación , Tamaño de la Partícula , Solubilidad , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Agua/química , Difracción de Rayos XRESUMEN
Gastric cancer (GC) is a kind of global malignancy. However, the expression pattern and clinical relevance of lamin B1 in GC remain to be elucidated. We endeavored to investigate how GC is influenced by lamin B1 and the related mechanisms. The lamin B1 expression in GC tissues from 71 patients was assessed by using immunohistochemistry (IHC). The expression of lamin B1 was connected with the clinical stage, depth of invasion, and poorer overall survival. Colony formation assays and methyl thiazolyl tetrazolium (MTT) were used to assess cell viability. The migration ability of GC cells was determined by cell scratch assay and Transwell invasion assay. Moreover, we used two cell lines of GC to explore the underlying mechanism of lamin B1 in boosting the GC cells proliferation and invasion in vitro by assessing the effects on related signal transduction pathways. Our data demonstrated that the expression level of lamin B1 was downregulated in GC tissues, and low expression level of lamin B1 was significantly correlated with higher clinical stage, depth of invasion, nodal stage, and poor prognosis. Moreover, in vitro experiments demonstrated that lamin B1 knockdown promoted, whereas lamin B1 overexpression inhibited, gastric cancer cell proliferation and migration. We also observed that lamin B1 knockdown could promote the activity of the PI3K/PTEN/Akt and MAPK/ERK pathway with a decrease in the p53/p21WAF1/CIP1 expression, whereas lamin B1 overexpression contributed to the opposite results. In conclusion, our studies indicate that lamin B1 deficiency is crucial in GC progression. Furthermore, the results elucidating the biological mechanisms of lamin B1 may potentially contribute to current GC treatment modalities.
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Lamina Tipo B/genética , Neoplasias Gástricas , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Pronóstico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Neoplasias Gástricas/genéticaRESUMEN
Objective: To investigate the efficacy of alternative donor (AD) in the treatment of aplastic anemia (AA) in children. Methods: The clinical data of AA children who received AD HSCT in our center from Apr. 2010 to Dec. 2016 were retrospectively analyzed. The overall survival (OS) rate, implant success rate, incidence of acute and chronic graft-versus-host disease (GVHD) were statistically analyzed. Results: A total of 109 children with acquired AA, including 64 severe AA (SAA) , 32 very severe AA (VSAA) and 13 transfusion dependent non-severe AA (NSAA) , were recruited in this retrospective AD HSCT study, the median age was 6 (0.8-18) years old. Of them, 44 patients with 10/10 matched unrelated donor (MUD) , 44 patients with mismatched unrelated donor (MMUD) and 21 patients with mismatched related donor (MMRD) . All patients did not receive ATG before HSCT and the active infection was excluded. Except 3 patients suffered from a second graft failure (2 of them rescued by second HSCT) , 106/109 (97.2%) were engrafted with neutrophil and platelet recovery occurring at a median of 13 days (range, 9-19) and 16 days (range, 10-81) post-transplant. Until day 100 post transplantation, the incidence was 74.3% (81/109) for acute GVHD (aGVHD) and 39.4% (43/109) for grade â ¡-â £ aGVHD, 30.7% (31/101) and 9.9% (10/101) for overall chronic GVHD (cGVHD) and moderate cGVHD, respectively, and nobody developed an extend cGVHD. After median follow up of 39 (0.7-103) months for all patients, 13 of 109 patients died. The estimated 5-year overall survival (OS) of the entire cohort was 88.1% (95%CI 81.1%-91.4%) with no difference among the MUD, MMUD and MMRD cohort (93.2%, 84.1% and 85.7%, respectively, P=0.361) . Conclusion: These excellent outcomes suggest that unmanipulated AD PBSC is a good HSCT source for children with SAA. It's reasonable to consider AD HSCT as first line therapy for SAA children without matched sibling donor. Better strategies are required to prevent GVHD.
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Anemia Aplásica , Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Adolescente , Niño , Preescolar , Humanos , Lactante , Estudios Retrospectivos , Donantes de Tejidos , Resultado del TratamientoRESUMEN
We develop a simple, fast and economical surface treatment under ambient temperature to improve the hydrophilicity and osteoconductivity of polyetheretherketone (PEEK) for bone implant applications. A major challenge in bone implants is the drastic difference in stiffness between traditional implant materials (such as titanium and stainless steel) and human bone. PEEK is biocompatible with an elastic modulus closely matching that of human bone, making it a highly attractive alternative. However, its bio-inert and poorly hydrophilic surface presents a serious challenge for osseointegration. Sulfonation can improve hydrophilicity and introduce bioactive sulfonate groups, but PEEK sulfonation has traditionally been applied for fuel cells, employing elevated temperatures and long reaction times to re-cast PEEK into sulfonated films. Little research has systematically studied PEEK surface modification by short reaction time (seconds) and ambient-temperature sulfonation for biomedical applications. Here, we investigate three ambient-temperature sulfonation treatments under varying reaction times (5-90 s) and evaluate the hydrophilicity and morphology of 15 modified PEEK surfaces. We establish an optimal treatment using 30 s H2SO4 followed by 20 s rinsing, and then 20 s immersion in NaOH followed by 20 s rinsing. This 30 s ambient-temperature sulfonation is found to be more effective than conventional plasma treatments and reduced PEEK water contact angle from 78° to 37°.
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Materiales Biocompatibles Revestidos/química , Calor , Cetonas/química , Oseointegración , Polietilenglicoles/química , Prótesis e Implantes , Benzofenonas , Humanos , Polímeros , Propiedades de SuperficieRESUMEN
Polymeric scaffolds comprising two size scales of microfibers and submicron fibers can better support three-dimensional (3D) cell growth in tissue engineering, making them an important class of healthcare material. However, a major manufacturing barrier hampers their translation into wider practical use: scalability. Traditional production of two-scale scaffolds by electrospinning is slow and costly. For day-to-day cell cultures, the scaffolds need to be affordable, made in high yield to drive down cost. Combining expertise from academia and industry from the United Kingdom and United States, this study uses a new series of high-yield, low-cost scaffolds made by shear spinning for tissue engineering. The scaffolds comprise interwoven submicron fibers and microfibers throughout as observed under scanning electron microscopy and demonstrate good capability to support cell culturing for tumor modeling. Three model human cancer cell lines (HEK293, A549 and MCF-7) with stable expression of GFP were cultured in the scaffolds and found to exhibit efficient cell attachment and sustained 3D growth and proliferation for 30 days. Cryosection and multiphoton fluorescence microscopy confirmed the formation of compact 3D cell clusters throughout the scaffolds. In addition, comparative growth curves of 2D and 3D cultures show significant cell-type-dependent differences. This work applies high-yield shear-spun scaffolds in mammalian tissue engineering and brings practical, affordable applications of multiscale scaffolds closer to reality. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2750, 2019.
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Resistencia al Corte , Proliferación Celular , Supervivencia Celular , Humanos , Microscopía Fluorescente , Ingeniería de Tejidos , Andamios del Tejido , Células Tumorales CultivadasRESUMEN
Objective: To investigate the peripheral small airway dysfunction differences between idiopathic pulmonary arterial hypertension (IPAH) and chronic thromboembolic pulmonary hypertension (CTEPH). Methods: Impulse oscillmetory system testing (IOS) and pulmonary function testing (PFT) were performed in IPAH and CTEPH patients and 30 healthy control group. We also carried out a subgroup analysis depending on their medical history of airway diseases. Results: We included 42 IPAH and 47 CTEPH patients (with or without airways disease: 8 vs. 34 and 17 vs. 34, respectively). Compared with CTEPH patients, IPAH patients were younger but had more serious pulmonary vessel resistance and mean pulmonary arterial resistance. Compared with IPAH patients, CTEPH patients had significant impaired peripheral small airway dysfunction with decreased of MEF(50) (% pred), MMEF(75/25) evaluated by PFT and R5-R20, Δ R5-R20 and AX measured by IOS [10.6(2.0, 33.0) vs. 2.5(-5.0, 16.5); 22.1(14.0, 32.6) vs. 15.5 (7.0, 23.2); 7.64(4, 18.6) vs. 6(3, 11) respectively, all P<0.05]. Subgroup analysis revealed there were no significant peripheral small dysfunction differences in IPAH patients with or without airway diseases. CTEPH patients had a higher proportion of airway diseases and more serious peripheral dysfunction than IPAH patients with airway diseases. Compared with control healthy group, peripheral airway dysfunction was more obvious even in IPAH and CTEPH patients without airway diseases. Conclusion: Compared with IPAH, CTEPH patients were older, but had better hemodynamics and a higher proportion of airway diseases. The peripheral airway dysfunction were more serious in CTEPH patients without airway diseases than IPAH patients without airway diseases and healthy controls group.
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Hipertensión Pulmonar Primaria Familiar , Hipertensión Pulmonar/diagnóstico , Embolia Pulmonar/diagnóstico , Tromboembolia/diagnóstico , Enfermedad Crónica , Humanos , Hipertensión Pulmonar/fisiopatología , Pulmón/fisiopatología , Arteria Pulmonar , Embolia Pulmonar/fisiopatología , Tromboembolia/fisiopatologíaRESUMEN
Electrospun nanofibrous scaffolds are promising regenerative wound dressing options but have yet to be widely used in practice. The challenge is that nanofibre productions rely on bench-top apparatuses, and the delicate product integrity is hard to preserve before reaching the point of need. Timing is critically important to wound healing. The purpose of this investigation is to produce novel nanofibrous scaffolds using a portable, hand-held "gun", which enables production at the wound site in a time-dependent fashion, thereby preserving product integrity. We select bacterial cellulose, a natural hydrophilic biopolymer, and polycaprolactone, a synthetic hydrophobic polymer, to generate composite nanofibres that can tune the scaffold hydrophilicity, which strongly affects cell proliferation. Composite scaffolds made of 8 different ratios of bacterial cellulose and polycaprolactone were successfully electrospun. The morphological features and cell-scaffold interactions were analysed using scanning electron microscopy. The biocompatibility was studied using Saos-2 cell viability test. The scaffolds were found to show good biocompatibility and allow different proliferation rates that varied with the composition of the scaffolds. A nanofibrous dressing that can be accurately moulded and standardised via the portable technique is advantageous for wound healing in practicality and in its consistency through mass production.
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Vendajes , Celulosa/uso terapéutico , Nanofibras/uso terapéutico , Poliésteres/uso terapéutico , Ingeniería de Tejidos/instrumentación , Ingeniería de Tejidos/métodos , Andamios del Tejido , Adulto , Anciano , Anciano de 80 o más Años , Proliferación Celular/fisiología , Supervivencia Celular/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Cicatrización de Heridas/fisiologíaRESUMEN
Objective: To improve the diagnosis and treatment of the pulmonary veno-occlusive disease (PVOD) and pulmonary capillary hemangioma (PCH). Methods: The clinical features, radiological findings, laboratory testing and treatment in 8 cases of PVOD/PCH which was diagnosed from 2013 to 2017 were described. Results: PVOD/PCH was rare. The clinical symptoms were easily confused with IPAH, but the decrease of hypoxemia, clubbing, D(L)CO were more obvious, and the imaging features of HRCT were helpful for PVOD/PCH diagnosis. Combined with gene testing, it was helpful to diagnose PVOD/PCH and avoid the risk of surgical biopsy. Conclusion: PVOD and PCH are rare type of pulmonary vascular diseases. According to clinical manifestations, physical examination, pulmonary function test results, HRCT imaging, CPET and gene detection results, PVOD or PCH can be diagnosed.
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Hemangioma Capilar/diagnóstico por imagen , Hipertensión Pulmonar , Neoplasias Pulmonares/patología , Pulmón/diagnóstico por imagen , Enfermedad Veno-Oclusiva Pulmonar/diagnóstico , Enfermedad Veno-Oclusiva Pulmonar/etiología , Hemangioma Capilar/patología , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Enfermedad Veno-Oclusiva Pulmonar/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVE: To observe the curative effect of dl-3-n-Butylphthalide (NBP) on patients with acute cerebral infarction (ACI) and its effects on levels of serum vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). PATIENTS AND METHODS: A total of 160 ACI patients treated in our hospital who met the criteria were selected and randomly divided into treatment group (n=80, including 42 males and 38 females) and control group (n=80, including 40 males and 40 females). The control group was treated with routine drug therapy, while the treatment group was treated with butylphthalide on this basis. The curative effect was evaluated using the National Institute of Health Stroke Scale (NIHSS) and the Activity of Daily Life Scale (ADL Scale). The levels of the two factors in serum were measured using enzyme-linked immunosorbent assay (ELISA), and the changes in the levels of the two factors in serum at different time points before and after treatment were compared between the two groups. RESULTS: After treatment, the levels of the two factors in serum in both groups were significantly increased compared with those before treatment (p<0.05), and the increase in treatment group was more significant than that in control group (p<0.05). The scores of ADL scale in both groups were significantly increased after treatment compared with those before treatment, and the increase in treatment group was more significant than that in control group (p<0.05). The scores of NIHSS in both groups were significantly decreased compared with those before treatment, and the decrease in treatment group was more significant than that in control group (p<0.05). CONCLUSIONS: NBP can improve the expressions of VEGF and bFGF in serum of ACI patients, and its effect is superior to that of conventional drugs.
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Benzofuranos/uso terapéutico , Infarto Cerebral/tratamiento farmacológico , Infarto Cerebral/metabolismo , Factor 2 de Crecimiento de Fibroblastos/biosíntesis , Fármacos Neuroprotectores/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Actividades Cotidianas , Adulto , Anciano , Anciano de 80 o más Años , Benzofuranos/efectos adversos , Infarto Cerebral/psicología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fármacos Neuroprotectores/efectos adversos , Accidente Cerebrovascular/tratamiento farmacológico , Accidente Cerebrovascular/psicología , Adulto JovenRESUMEN
BACKGROUND: Cell culture methods allow the detailed observations of individual plant cells and their internal processes. Whereas cultured cells are more amenable to microscopy, they have had limited use when studying the complex interactions between cell populations and responses to external signals associated with tissue and whole plant development. Such interactions result in the diverse range of cell shapes observed in planta compared to the simple polygonal or ovoid shapes in vitro. Microfluidic devices can isolate the dynamics of single plant cells but have restricted use for providing a tissue-like and fibrous extracellular environment for cells to interact. A gap exists, therefore, in the understanding of spatiotemporal interactions of single plant cells interacting with their three-dimensional (3D) environment. A model system is needed to bridge this gap. For this purpose we have borrowed a tool, a 3D nano- and microfibre tissue scaffold, recently used in biomedical engineering of animal and human tissue physiology and pathophysiology in vitro. RESULTS: We have developed a method of 3D cell culture for plants, which mimics the plant tissue environment, using biocompatible scaffolds similar to those used in mammalian tissue engineering. The scaffolds provide both developmental cues and structural stability to isolated callus-derived cells grown in liquid culture. The protocol is rapid, compared to the growth and preparation of whole plants for microscopy, and provides detailed subcellular information on cells interacting with their local environment. We observe cell shapes never observed for individual cultured cells. Rather than exhibiting only spheroid or ellipsoidal shapes, the cells adapt their shape to fit the local space and are capable of growing past each other, taking on growth and morphological characteristics with greater complexity than observed even in whole plants. Confocal imaging of transgenic Arabidopsis thaliana lines containing fluorescent microtubule and actin reporters enables further study of the effects of interactions and complex morphologies upon cytoskeletal organisation both in 3D and in time (4D). CONCLUSIONS: The 3D culture within the fibre scaffolds permits cells to grow freely within a matrix containing both large and small spaces, a technique that is expected to add to current lithographic technologies, where growth is carefully controlled and constricted. The cells, once seeded in the scaffolds, can adopt a variety of morphologies, demonstrating that they do not need to be part of a tightly packed tissue to form complex shapes. This points to a role of the immediate nano- and micro-topography in plant cell morphogenesis. This work defines a new suite of techniques for exploring cell-environment interactions.
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Arabidopsis/crecimiento & desarrollo , Técnicas de Cultivo de Célula/métodos , Morfogénesis , Células Vegetales/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Células Cultivadas , Andamios del TejidoRESUMEN
The availability of forming technologies able to mass produce porous polymeric microspheres with diameters ranging from 150 to 300 µm is significant for some biomedical applications where tissue augmentation is required. Moreover, appropriate assembly of microspheres into scaffolds is an important challenge to enable direct usage of the as-formed structures in treatments. This work reports the production of poly (glycolic-co-lactic acid) and poly (ε-caprolactone) microspheres under ambient conditions using one-step electrohydrodynamic jetting (traditionally known as atomisation) and thermally induced phase separation (TIPS). To ensure robust production for practical uses, this work presents 12 comprehensive parametric mode mappings of the diameter distribution profiles of the microspheres obtained over a broad range of key processing parameters and correlating of this with the material parameters of 5 different polymer solutions of various concentrations. Poly (glycolic-co-lactic acid) (PLGA) in Dimethyl carbonate (DMC), a low toxicity solvent with moderate conductivity and low dielectric constant, generated microspheres within the targeted diameter range of 150-300 µm. The fabrication of the microspheres suitable for formation of the scaffold structure is achieved by changing the collection method from distilled water to liquid nitrogen and lyophilisation in a freeze dryer.
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Microesferas , Tamaño de la Partícula , Polímeros/químicaRESUMEN
Academic and industrial research on nanofibres is an area of increasing global interest, as seen in the continuously multiplying number of research papers and patents and the broadening range of chemical, medical, electrical and environmental applications. This in turn expands the size of the market opportunity and is reflected in the significant rise of entrepreneurial activities and investments in the field. Electrospinning is probably the most researched top-down method to form nanofibres from a remarkable range of organic and inorganic materials. It is well known and discussed in many comprehensive studies, so why this review? As we read about yet another "novel" method producing multifunctional nanomaterials in grams or milligrams in the laboratory, there is hardly any research addressing how these methods can be safely, consistently and cost-effectively up-scaled. Despite two decades of governmental and private investment, the productivity of nanofibre forming methods is still struggling to meet the increasing demand. This hinders the further integration of nanofibres into practical large-scale applications and limits current uses to niche-markets. Looking into history, this large gap between supply and demand of synthetic fibres was seen and addressed in conventional textile production a century ago. The remarkable achievement was accomplished via extensive collaborative research between academia and industry, applying ingenious solutions and technological convergence from polymer chemistry, physical chemistry, materials science and engineering disciplines. Looking into the present, current advances in electrospinning and nanofibre production are showing similar interdisciplinary technological convergence, and knowledge of industrial textile processing is being combined with new developments in nanofibre forming methods. Moreover, many important parameters in electrospinning and nanofibre spinning methods overlap parameters extensively studied in industrial fibre processing. Thus, this review combines interdisciplinary knowledge from the academia and industry to facilitate technological convergence and offers insight for upscaling electrospinning and nanofibre production. It will examine advances in electrospinning within a framework of large-scale fibre production as well as alternative nanofibre forming methods, providing a comprehensive comparison of conventional and contemporary fibre forming technologies. This study intends to stimulate interest in addressing the issue of scale-up alongside novel developments and applications in nanofibre research.
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Equipos y Suministros Eléctricos , Nanofibras/química , Nanotecnología/instrumentación , Diseño de Equipo , Historia del Siglo XX , Nanofibras/ultraestructura , Nanotecnología/métodos , Textiles/historiaRESUMEN
A series of 41 papers by H. Zhong et al. are retracted.
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AIMS: To examine the expression of ADAM12 (meltrin alpha), a member of the disintegrin and metalloprotease (ADAM) family, in human giant cell tumours of the bone, skeletal muscle tissue from human embryos, and human adult skeletal muscle tissue. METHODS: ADAM12 mRNA was detected by reverse transcription polymerase chain reaction and in situ hybridisation. RESULTS: ADAM12 mRNA was detected in 14 of the 20 giant cell tumours of bone and in three of the six tumour cell cultures. The expression of ADAM12 in cells cultured from the tumour was linked to the presence of multinucleated giant cells. ADAM12 mRNA could not be detected in the five adult skeletal muscle tissue samples, although it was found in the two embryonic skeletal muscle tissue samples. ADAM12 mRNA was localised to the cytoplasm of multinucleated giant cells and some mononuclear stromal cells. CONCLUSIONS: These results indicate that multinucleated giant cells are formed by the cell fusion of mononuclear stromal cells in giant cell tumours of bone and that ADAM12 is involved in the cell fusion process.