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Toxocara canis is a parasitic zoonose that is distributed worldwide and is one of the two pathogens causing toxocariasis. After infection, it causes serious public health and safety problems, which pose significant veterinary and medical challenges. To better understand the regulatory effects of T. canis infection on the host immune cells, murine macrophages (RAW264.7) were incubated with recombinant T. canis C-type lectin 4 (rTc-CTL-4) protein in vitro. The quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the nucleotide-binding oligomerization domain-containing protein 1/2 (NOD1/2), receptor-interacting protein 2 (RIP2), nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase (MAPK) on mRNA level and protein expression level in macrophages. Our results indicated that 10 µg/mL rTc-CTL-4 protein could modulate the expression of NOD1, NOD2, and RIP2 at both the transcriptional and translational levels. The protein translation levels of NF-κB, P-p65, p38, and P-p38 in macrophages were also modulated by rTc-CTL-4 protein. Macrophages were co-incubated with rTc-CTL-4 protein after siRNA silencing of NOD1, NOD2, and RIP2. The expression levels of NF-κB, P-p65, p38, and P-p38 were significantly changed compared with the negative control groups (Neg. Ctrl.). Taken together, rTc-CTL-4 protein seemed to act on NOD1/2-RIP2-NF-κB and MAPK signaling pathways in macrophages and might activate MAPK and NF-κB signaling pathways by regulating NOD1, NOD2, and RIP2. The insights from the above studies could contribute to our understanding of immune recognition and regulatory mechanisms of T. canis infection in the host animals.
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FN-kappa B , Toxocara canis , Animales , Ratones , FN-kappa B/genética , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Toxocara canis/metabolismo , Transducción de Señal/fisiología , MacrófagosRESUMEN
It remains a significant challenge in prosthetic rehabilitation for combined hard and soft palate defects on account of two primary reasons. At first, conventional impressions can hardly get an accurate analogue and usually bring about a terrible experience for the patients. Secondly, conventional hard denture base resins used in obturator prostheses exhibit limitations in marginal sealing, undercut retention, and elastic buffering when in contact with the soft palate. This article presents a case where combined hard and soft palate defects were successfully and rapidly reconstructed by using digital intraoral impression technology and denture soft reline material.
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Urinary incontinence (UI) is known as a distressing condition particularly among older adults, and negatively associated with health-related quality of life in both males and females. Prelamin A accumulation has been found in all progeroid laminopathies and is obviously linked to cell and organism aging. Therefore, this study was expected to investigate the effect of prelamin A on detrusor on UI. Prelamin A expression in clinical and animal samples was detected. To investigate the degree of prelamin A accumulation and detrusor calcification/aging, the detrusor cells were subcultured separately into low and high passage. The low-passage subculture cells were treated with transfection of overexpressed prelamin A plasmid, and transfection of overexpressed prelamin A plasmid and application of farnesyl transferase inhibitor (FTIs) H-9279, respectively. Zmpste24, Icmt and lamin A/C expression were detected to explore how prelamin A affected detrusor calcification/aging. Prelamin A was overexpressed in aged detrusor cells, indicating prelamin A expression was positively related to the age of subjects. The degree of prelamin A accumulation and detrusor calcification/aging was higher in aged rats and high passage subculture cells. Zmpste24, Icmt and lamin A/C were poorly expressed in cells transfected with overexpressed prelamin A, as well as cell proliferation activity decreased and calcium deposition and apoptotic rate increased. Furthermore, we also found that the effect of overexpressed prelamin A was lost when cells were treated with H-9279. These findings provide evidence that prelamin A overexpression impairs degradation of its farnesylated form, thus causing prelamin A accumulation which induces detrusor calcification/aging in UI.
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Envejecimiento/metabolismo , Calcinosis/metabolismo , Lamina Tipo A/metabolismo , Incontinencia Urinaria/metabolismo , Adulto , Anciano , Animales , Células Cultivadas , Femenino , Humanos , Proteínas de Filamentos Intermediarios/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Metaloendopeptidasas/metabolismo , Proteínas Nucleares/metabolismo , Calidad de Vida , Ratas , Ratas Sprague-DawleyRESUMEN
Long noncoding RNAs (lncRNAs) were proposed to be important regulators influencing various differentiation processes. Yet, the molecular mechanisms of lncRNAs governing osteogenic differentiation of Periodontal Ligament Stem Cells (PDLSCs) remain unclear. Here, PDLSCs were isolated from normal periodontal ligament of human (PDL) whereas P-PDLSCs were isolated from periodontitis affected PDL. Quantitative real-time PCR (qRT-PCR) was performed to examine the relative expression level of lncRNA-ANCR and of Osterix (OSX), Alkaline Phosphatase (ALP) as well as Runt-related transcription factor 2 (RUNX2) in PDLSCs. Gain- and loss-of- function experiments was performed to study the role of lncRNA-ANCR. Alizarin Red staining was used to evaluate the function of lncRNA-ANCR and miRNA-758 on osteogenic differentiation. In addition, via dual luciferase reporter assay and RNA immunoprecipitation the microRNA sponge potential of lncRNA-ANCR was assessed. A luciferase reporter assay identified the correlation between miR-758 and Notch2. Our results showed that the expression of ALP, RUNX2 and OSX were increased whereas lncRNA-ANCR was decreased during the process of differentiation in PDLSCs. Overexpression of lncRNA-ANCR decreased the expression of ALP, RUNX2 and OSX as confirmed by Alizarin red staining. Overexpression of lncRNA-ANCR resulted in reduction of the miR-758 expression level. Furthermore, RNA immunoprecipitation proved that lncRNA-ANCR targets miR-758 directly. The results of dual luciferase reporter assay also demonstrated that miR-758 regulated Notch2 expression by targeting 3'-UTR of Notch2. In conclusion, the novel pathway lncRNA-ANCR/miR-758/Notch2 plays an important role in the process of regulating osteogenic differentiation of PDLSCs.
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Regulación de la Expresión Génica , MicroARNs/genética , Osteogénesis/genética , Ligamento Periodontal/citología , ARN Largo no Codificante/genética , Células Madre/metabolismo , Regiones no Traducidas 3'/genética , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Secuencia de Bases , Diferenciación Celular/genética , Humanos , Receptor Notch2/genética , Receptor Notch2/metabolismo , Homología de Secuencia de Ácido Nucleico , Factor de Transcripción Sp7/genética , Factor de Transcripción Sp7/metabolismo , Vía de Señalización Wnt/genéticaRESUMEN
Toxocara canis is a zoonotic parasite with worldwide distribution. ATP-binding cassette (ABC) transporters are integral membrane proteins which involve in a range of biological processes in various organisms. In present study, the full-length coding sequence of abcg-5 gene of T. canis (Tc-abcg-5) was cloned and characterized. A 633 aa polypeptide containing two conserved Walker A and Walker B motifs was predicted from a continuous 1902 nt open reading frame. Quantitative real-time PCR was employed to determine the transcriptional levels of Tc-abcg-5 gene in adult male and female worms, which indicated high mRNA level of Tc-abcg-5 in the reproductive tract of adult female T. canis. Tc-abcg-5 was expressed to produce rabbit polyclonal antiserum against recombinant TcABCG5. Indirect-fluorescence immunohistochemical assays were carried out to detect the tissue distribution of TcABCG5, which showed predominant distribution of TcABCG5 in the uterus (especially in the germ cells) of adult female T. canis. Tissue transcription and expression pattern of Tc-abcg-5 indicated that Tc-abcg-5 might play essential roles in the reproduction of this parasitic nematode.
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Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 5/biosíntesis , Toxocara canis/genética , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 5/genética , Animales , Enfermedades de los Perros/parasitología , Perros , Femenino , Masculino , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducción , Distribución Tisular , Toxocara canis/aislamiento & purificación , Toxocara canis/fisiología , Toxocariasis/parasitología , Transcripción Genética , Útero/metabolismoRESUMEN
Toxocara canis is an common intestinal nematode of canids and the principal causative agent of human toxocariasis. Vitellogenin (Vg), a source of amino acids and lipids in the eggs, are considered to play an important role in embryo development of a wide range of organisms. In the present study, the transcriptional levels of Tc-vit-6 gene in male and female adult T. canis were determined by quantitative real-time PCR, which indicated high transcription of Tc-vit-6 in the intestine, reproductive tract and body wall of male and female adult T. canis. The fragment of Tc-vit-6 encoding a vWD domain, was cloned and expressed to produce a rabbit anti-TcvWD polyclonal antibody. Tissue distribution of TcVg6 was detected by immunohistochemical assays, which showed predominant distribution of TcVg6 in the tissues of intestine, as well as reproductive tract (including some of the germ cells) and musculature of male and female adult worms. Collectively, these results indicated multiple biological roles of TcVg6 apart from that in the reproduction of T. canis.
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Toxocara canis/metabolismo , Toxocariasis/parasitología , Vitelogeninas/metabolismo , Animales , Anticuerpos Antihelmínticos/biosíntesis , Western Blotting , Canidae/parasitología , Perros , Femenino , Regulación de la Expresión Génica , Genitales/metabolismo , Humanos , Inmunohistoquímica , Mucosa Intestinal/metabolismo , Masculino , Músculos/metabolismo , Conejos , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Distribución Tisular , Transcripción Genética , Vitelogeninas/genética , Vitelogeninas/inmunología , Vitelogeninas/fisiologíaRESUMEN
Background: This study evaluated the efficiency of corticosteroid, leflunomide and mesenchymal stem cells (MSCs) in the treatment of pediatric idiopathic pulmonary hemosiderosis (IPH). Methods: Ten patients were included in the study. The diagnosis of IPH was based on clinical symptoms, laboratory examinations and pulmonary hemosiderosis. Induction therapy consisted of methylprednisolone pulse therapy, followed by prednisone plus leflunomide. Maintenance therapy consisted of low-dose prednisone, leflunomide and administration of MSCs. Results: All the patients achieved complete response after treatment with corticosteroid, leflunomide and MSCs. The median follow-up was 23 months (range: 4-34 months). Moreover, administration of MSCs induced an increase in the percentage of CD4+ CD25+ regulatory T cells but a decrease in the percentage of Th17 cells. Conclusion: Treatment with corticosteroid, leflunomide and MSCs for pediatric IPH was safe and effective.
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Corticoesteroides/uso terapéutico , Hemosiderosis/terapia , Inmunosupresores/uso terapéutico , Isoxazoles/uso terapéutico , Enfermedades Pulmonares/tratamiento farmacológico , Trasplante de Células Madre Mesenquimatosas , Niño , Femenino , Glucocorticoides/administración & dosificación , Glucocorticoides/uso terapéutico , Hemosiderosis/diagnóstico , Humanos , Leflunamida , Enfermedades Pulmonares/diagnóstico , Enfermedades Pulmonares/terapia , Masculino , Células Madre Mesenquimatosas , Metilprednisolona/uso terapéutico , Prednisona/administración & dosificación , Quimioterapia por Pulso , Estudios Retrospectivos , Resultado del Tratamiento , Hemosiderosis PulmonarRESUMEN
Toxocara canis is an intestinal nematode of canids with a worldwide distribution, causing an important but neglected parasitic zoonosis in humans. Aquaporins (AQP) are a family of water channel proteins, which function as membrane channels to regulate water homeostasis. In this study, the coding sequence of aquaporin-1 gene of T. canis (Tc-aqp-1) was cloned and characterized. The obtained Tc-aqp-1 coding sequence was 933 bp in length, which predicted to encode 311 amino acids. Two conserved asparagine-proline-alanine (NPA) motifs were identified in the multiple sequence alignments. Phylogenetic analysis revealed the closest relationship between T. canis and Opisthorchis viverrini based on aquaporin-1 amino acid sequence. A structure was predicted with ligand binding sites predicted at H93, N95, N226, L94, I79, and I210 and with active sites predicted at I256 and G207. Gene Ontology (GO) annotations predicted its cellular component term of integral component of plasma membrane (GO: 0005887), molecular function term of channel activity (GO: 0015250), and biological process term of water transport (GO: 0006833). Tissue expression analysis revealed that the Tc-aqp-1 was highly expressed in the intestine of adult male. The findings of the present study provide the basis for further functional studies of T. canis aquaporin-1.
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Acuaporina 1/genética , Toxocara canis/genética , Secuencia de Aminoácidos , Animales , Acuaporina 1/química , Femenino , Humanos , Masculino , Oligopéptidos/química , Opisthorchis/clasificación , Opisthorchis/genética , Filogenia , Alineación de Secuencia , Toxocara canis/clasificaciónRESUMEN
Toxocara canis (T. canis) is a widely prevalent zoonotic parasite that infects a wide range of mammalian hosts, including humans. We generated the full-length complementary DNA (cDNA) of the serine/threonine phosphatase gene of T. canis (Tc stp) using 5' rapid amplification of the cDNA ends. The 1192-bp sequence contained a continuous 942-nucleotide open reading frame, encoding a 313-amino-acid polypeptide. The Tc STP polypeptide shares a high level of amino-acid sequence identity with the predicted STPs of Loa loa (89%), Brugia malayi (86%), Oesophagostomum columbianum (76%), and Oesophagostomumdentatum (76%). The Tc STP contains GDXHG, GDXVDRG, GNHE motifs, which are characteristic of members of the phosphoprotein phosphatase family. Our quantitative real-time polymerase chain reaction analysis showed that the Tc STP was expressed in six different tissues in the adult male, with high-level expression in the spermary, vas deferens, and musculature, but was not expressed in the adult female, suggesting that Tc STP might be involved in spermatogenesis and mating behavior. Thus, STP might represent a potential molecular target for controlling T. canis reproduction.
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ADN Complementario/química , Fosfoproteínas Fosfatasas/genética , ARN de Helminto/genética , Toxocara canis/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Perros , Femenino , Masculino , Datos de Secuencia Molecular , Fosfoproteínas Fosfatasas/química , Fosfoproteínas Fosfatasas/metabolismo , Filogenia , ARN de Helminto/aislamiento & purificación , Alineación de Secuencia , Toxocara canis/clasificación , Toxocara canis/genéticaRESUMEN
RAF/MEK/ERK pathway is frequently activated in tumor. Therefore, this study will investigate the function of RUVBL1 (RAF-binding protein) in tongue squamous cell carcinoma (TSCC). Bioinformatics was performed to identify differentially expressed mRNAs (DE-mRNAs) in TCGA-oral squamous cell carcinoma, GSE13601, and GSE34105 datasets. A total of 672 shared DE-mRNAs were identified in three datasets, and they are regulating metastasis and angiogenesis. Patients with RUVBL1 low expression had high overall survival. Overexpressing RUVBL1 enhanced the viability, wound healing percentage, invasion, sphere formation, angiogenesis, and resistance to cisplatin and 5-fluorouracil in CAL-27 and SCC-4 cells, and the opposite results were obtained by knocking down RUVBL1. Moreover, overexpression of RUVBL1 bolstered tumor growth in vivo. Strikingly, RUVBL1 diminished the phosphorylation of CRAF Ser259, which led to activation of the MEK/ERK pathway. In conclusion, RUVBL1 contributes to the malignant biological behavior of TSCC via activating the CRAF/MEK/ERK pathway. This provides molecular mechanisms and perspectives for targeted therapy of the CRAF/MEK/ERK pathway.
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Low viability of seed cells and the concern about biosafety restrict the application of cell-based tissue-engineered bone (TEB). Exosomes that bear similar bioactivities to donor cells display strong stability and low immunogenicity. Human umbilical cord mesenchymal stem cells-derived exosomes (hUCMSCs-Exos) show therapeutic efficacy in various diseases. However, little is known whether hUCMSCs-Exos can be used to construct TEB to repair bone defects. Herein, PM-Exos and OM-Exos were separately harvested from hUCMSCs which were cultured in proliferation medium (PM) or osteogenic induction medium (OM). A series of in-vitro studies were performed to evaluate the bioactivities of human bone marrow mesenchymal stem cells (hBMSCs) when co-cultured with PM-Exos or OM-Exos. Differential microRNAs (miRNAs) between PM-Exos and OM-Exos were sequenced and analyzed. Furthermore, PM-Exos and OM-Exos were incorporated in 3D printed tricalcium phosphate scaffolds to build TEBs for the repair of critical-sized calvarial bone defects in rats. Results showed that PM-Exos and OM-Exos bore similar morphology and size. They expressed representative surface markers of exosomes and could be internalized by hBMSCs to promote cellular migration and proliferation. OM-Exos outweighed PM-Exos in accelerating the osteogenic differentiation of hBMSCs, which might be attributed to the differentially expressed miRNAs. Furthermore, OM-Exos sustainably released from the scaffolds, and the resultant TEB showed a better reparative outcome than that of the PM-Exos group. Our study found that exosomes isolated from osteogenically committed hUCMSCs prominently facilitated the osteogenic differentiation of hBMSCs. TEB grafts functionalized by OM-Exos bear a promising application potential for the repair of large bone defects.
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Mandibular angle osteotomy is commonly used for prominent mandibular angle contouring. Because of difficult control of the line, shape, and amount of osteotomy, many complications such as asymmetry, undercorrection, overcorrection, and formation of a second mandibular angle after surgery occurred commonly. In addition, it is more difficult to implement osteotomy exactly the same as preoperative design in curved osteotomy owing to the arc-shaped osteotomy line. Therefore, further studies are needed to explore ways to make osteotomy accurately identical with preoperative design. In this report, a case of curved osteotomy guided by a stereolithographic template for unilateral prominent mandibular angle is described. We established the osteotomy line, the shape, and the volume with Mimics software for the right prominent mandibular angle; fabricated individual osteotomy template with computer-aided design/computer-aided manufacturing (CAD/CAM) technique; and performed curved osteotomy with the template. A secure fit of the template on the bone surface was found during the operation. Computed tomographic scan after the surgery revealed that bilateral mandibular angles were symmetric, that the right mandibular angle had a natural curve, and that the osteotomy line, shape, and amount were in accordance with the preoperative design. It is suggested that CAD/CAM template could guide curved osteotomy for prominent mandibular angle accurately, improve efficiency, and avoid complications in osteotomy.
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Diseño Asistido por Computadora , Asimetría Facial/cirugía , Mandíbula/cirugía , Osteotomía/instrumentación , Estética , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Masculino , Modelos Anatómicos , Tempo Operativo , Planificación de Atención al Paciente , Satisfacción del Paciente , Tomografía Computarizada Espiral/métodos , Interfaz Usuario-Computador , Adulto JovenRESUMEN
Scaffolds prepared by 3D printing are increasingly used in the field of bone tissue repair. However, on traditional 3D printed bone tissue engineering scaffolds, cells can only grow on the fiber surface and form bone. We designed a scaffold with a cross-scale structure of PCL/ß-TCP, which contains thick fibers with a diameter of 500 µm printed by FDM. And in the pores of the coarse fiber, the ultra-high precision fine fiber grid with a diameter of about 10 µm is filled by MEW mode. In cell experiments, cells can not only grow on the thick fiber surface of the cross-scale scaffold. At the same time, the mesh structure of fine fibers provides a bridge for cell growth, allowing cells to pass through the pores of thick fibers and grow in the pores and gradually cover the pores of the scaffold. In the osteoinduction experiment, ß-TCP in the PCL/ß-TCP composite provides Ca2+ and PO43- to the scaffold, which effectively promotes the osteogenic differentiation of cells on the scaffold. Compared with traditional scaffolds, the osteogenic performance of cross-scale scaffolds is greatly improved. Not only did bone form on the surface of the scaffold, but also obvious ALP expression and effective calcium precipitation appeared in the pores of the scaffold. This can effectively speed up the repair of bone defects. We believe that the 3D printed PCL/ß-TCP cross-scale scaffold with high-precision fibers has great application prospects in the field of bone tissue engineering.
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Osteogénesis , Poliésteres , Huesos , Fosfatos de Calcio , Impresión Tridimensional , Ingeniería de Tejidos , Andamios del TejidoRESUMEN
OBJECTIVE: To investigate the short-term efficacy and safety of generic bortezomib in the treatment of Chinese patients with multiple myeloma (MM). METHODS: Clinical data of 62 MM patients (median age of 62 years) who had accepted at least 2 cycles of chemotherapy based on generic bortezomib in our center from December 2017 to July 2019 were retrospectively analyzed, including 47 newly diagnosed patients and 15 with disease recurrence or progression. RESULTS: Anemia, renal dysfunction, hypoproteinemia and high level of ß 2-microglobulin were all improved rapidly after induction treatment. In 56 patients who had completed at least 4 cycles of chemotherapy, the overall response rate (ORR) was 85.7%, and 64.3% of the patients achieved very good partial response (VGPR) or better, and 28.6% achieved complete remission (CR) or better. In the 19 patients who had already completed all planned induction and consolidation treatment (9 cycles of chemotherapy or 4-6 cycles of chemotherapy plus autologous hematopoietic stem cell transplantation), 84.2% achieved VGPR or better, and 57.9% achieved CR or stringent complete remission (sCR). Median follow-up time was 300 days with data cut-off date of September 20, 2019, and the progression-free survival (PFS) rate and overall survival (OS) rate were 62.1% and 85.3%, respectively. The possible adverse reactions associated with bortezomib were grade 1-2, the most common hematologic adverse reaction was thrombocytopenia (27.4%), and the most common non-hematologic adverse reaction was peripheral neuropathy (43.5%), followed by asthenia (37.1%). CONCLUSION: The disease severity can be rapidly alleviated after generic bortezomib-based chemotherapy, and a favorable short-term efficacy and survival have been observed with a generally acceptable toxicity profile. However, the long-term outcomes will be examined through further follow-up.
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Mieloma Múltiple , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Bortezomib/uso terapéutico , Dexametasona/uso terapéutico , Supervivencia sin Enfermedad , Humanos , Persona de Mediana Edad , Mieloma Múltiple/tratamiento farmacológico , Recurrencia Local de Neoplasia , Estudios Retrospectivos , Trasplante Autólogo , Resultado del TratamientoRESUMEN
OBJECTIVES: Mesenchymal stem cells (MSCs) based therapy for bone regeneration has been regarded as a promising method in the clinic. However, hBMSCs with invasive harvesting process and undesirable proliferation rate hinder the extensive usage. HUCMSCs of easier access and excellent performances provide an alternative for the fabrication of tissue-engineered bone construct. Evidence suggested the osteogenesis ability of hUCMSCs was weaker than that of hBMSCs. To address this issue, a co-culture strategy of osteogenically and angiogenically induced hUCMSCs has been proposed since thorough vascularization facilitates the blood-borne nutrition and oxygen to transport in the scaffold, synergistically expediting the process of ossification. MATERIALS AND METHODS: Herein, we used osteogenic- and angiogenic-differentiated hUCMSCs for co-culture in screened culture medium to elevate the osteogenic capacity with in vitro studies and finally coupled with 3D TCP scaffold to repair rat's critical-sized calvarial bone defect. By dual-directional induction, hUCMSCs could differentiate into osteoblasts and endothelial cells, respectively. To optimize the co-culture condition, gradient ratios of dual-directional differentiated hUCMSCs co-cultured under different medium were studied to determine the appropriate condition. RESULTS: It revealed that the osteogenic- and angiogenic-induced hUCMSCs mixed with the ratio of 3:1 co-cultured in the mixed medium of osteogenic induction medium to endothelial cell induction medium of 3:1 possessed more mineralization nodules. Similarly, ALP and osteogenesis/angiogenesis-related genes expressions were relatively higher. Further evidence of bone defect repair with 3D printed TCP of 3:1 group exhibited better restoration outcomes. CONCLUSIONS: Our work demonstrated a favourable and convenient approach of dual-directional differentiated hUCMSCs co-culture to improve the osteogenesis, establishing a novel way to fabricate tissue-engineered bone graft with 3D TCP for large bone defect augmentation.
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Técnicas de Cocultivo/métodos , Células Madre Mesenquimatosas/citología , Osteogénesis/fisiología , Cordón Umbilical/citología , Animales , Regeneración Ósea/fisiología , Huesos/citología , Diferenciación Celular/fisiología , Células Cultivadas , Células Endoteliales/citología , Humanos , Masculino , Osteoblastos/citología , Ratas , Ratas Sprague-Dawley , Ingeniería de Tejidos/métodos , Andamios del TejidoRESUMEN
BACKGROUND: Circular RNAs (circRNAs) comprise a novel class of noncoding RNAs that play important roles in a variety of diseases. However, the mechanism by which circRNAs regulate the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs) remains largely unclear. METHODS: Microarray analysis was used to explore the expression profiles of circRNAs during the osteogenic differentiation of normal and BMP2 induced-MSMSCs. CircRNA_33287 was identified by agarose electrophoresis, quantitative real-time PCR (qRT-PCR), and western blotting. The function of circRNA_33287 was assessed by loss- and gain-of-function techniques and Alizarin red staining. Potential miRNA binding sites for circRNA_33287, and the target genes of miR-214-3p, were predicted by using online bioinformatics analysis tools. The relationships among the regulatory roles played by circRNA_33287, miR-214-3p, and Runt-related transcription factor 3 (Runx3), during the osteogenic differentiation of MSMSCs were verified by use of the dual luciferase reporter assay, qRT-PCR, and western blotting techniques, respectively. In addition, the molecular sponge potential of circRNA_33287 for miRNA was assessed via in vivo ectopic bone formation and a histological analysis performed after hematoxylin and eosin staining. RESULTS: Expression of circRNA_33287 was confirmed to be up-regulated during the osteogenic differentiation of MSMSCS. Overexpression and silencing of circRNA_33287 increased and decreased the expression levels of key markers of osteogenesis, respectively, including Runx2, OSX, and ALP. Furthermore, circRNA_33287 acted as a molecular sponge for miR-214-3p, which regulated Runx3 expression by targeting its 3'UTR. Moreover, circRNA_33287 protected Runx3 from miR-214-3p-mediated suppression. In addition, circRNA_33287 was shown to increase ectopic bone formation in vivo and displayed the strongest ability to stimulate bone formation when co-transfected with a miR-214-3p inhibitor. CONCLUSION: The novel pathway circRNA_33287/miR-214-3p/Runx3 was found to play a role in regulating the osteoblastic differentiation of MSMSCs in the posterior maxilla.
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Subunidad alfa 3 del Factor de Unión al Sitio Principal/biosíntesis , Seno Maxilar/metabolismo , MicroARNs/biosíntesis , Osteogénesis/fisiología , ARN/biosíntesis , Células Madre/metabolismo , Animales , Diferenciación Celular/fisiología , Células Cultivadas , Células HEK293 , Humanos , Masculino , Seno Maxilar/citología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , ARN Circular , ARN Largo no Codificante/biosíntesisRESUMEN
We wished to investigate the role of a tilapia skin collagen polypeptide (TSCP; molecular weight <3â¯kDa) in alleviating liver and kidney injuries in aging mice induced by d-galactose (d-gal) and its underlying mechanism of action. First, we characterized TSCP. TSCP was passed through a 3-kDa ultrafiltration membrane, desalted in water by a solid-phase extraction column, purified further by reverse phase-high performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry and tandem mass spectrometry. TSCP contained 17 types of amino acids (AAs) and 41 peptide chains of length 7 AAs to 22 AAs. The content of free AAs and total AAs of TSCP was 13.5% and 93.79%, respectively. Next, we undertook animal experiments. Mice were injected once-daily with D-gal (300â¯mg/kg body weight, s.c.) for 8 weeks, and TSCP was administered simultaneously once-daily by intragastric gavage. TSCP could visibly improve the decreased body weight, depressed appetite, and mental deterioration of mice triggered by d-gal. TSCP could also alleviate d-gal-induced damage to the liver and kidneys according to histopathology (especially high-dose TSCP). Consistent with these macroscopic and pathologic changes, TSCP could also prevent d-gal-induced increases in serum levels of alanine aminotransferase, aspartate transaminase, alkaline phosphatase, lipid peroxidation, creatinine and uric acid, as well as decreases in serum levels of immunoglobulin (Ig)G and IgM. Moreover, TSCP improved the activities of superoxide dismutase, catalase, and glutathione peroxidase, but also inhibited the increases in the levels of malondialdehyde and inducible nitric oxide synthase expression in the liver and kidneys of d-gal-treated mice. These results suggest that TSCP can alleviate the injuries to the liver and kidneys in aging mice induced by d-gal, and that its mechanism of action might be, at least partially, associated with attenuation of oxidative stress and enhancement of immune function.
Asunto(s)
Colágeno/farmacología , Galactosa/efectos adversos , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Péptidos/farmacología , Sustancias Protectoras/farmacología , Tilapia/metabolismo , Alanina Transaminasa/metabolismo , Animales , Antioxidantes/metabolismo , Aspartato Aminotransferasas/metabolismo , Catalasa/metabolismo , Glutatión Peroxidasa/metabolismo , Riñón/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/metabolismo , Masculino , Malondialdehído/metabolismo , Ratones , Estrés Oxidativo/efectos de los fármacos , Superóxido Dismutasa/metabolismoRESUMEN
Surface water samples were collected in different seasons from Chaohu Lake to determine the concentrations and potential environmental impacts of nine dissolved heavy metals (As, Hg, Cd, Cu, Pb, Zn, Cr, Ni, and Co). The concentrations of the selected heavy metals were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES, ICAP6000 series). The single pollution index method and integrated pollution index method were used to evaluate the potential environmental impacts. The results indicated that the contents of dissolved As, Cd, Pb, and Cr in Chaohu Lake were lower than the limitations of Gradeâ of the Environmental Quality Standards for Surface Water (GB 3838-2002), while Cu and Zn were within the limitations of Gradeâ -â ¡ and Hg was within the limitations of Gradeâ -â ¢. The contents of Ni and Co were far below the limitations. The concentrations of the selected heavy metals (Cu, Pb, Zn, Cr, Ni, and Co) varied among seasons and areas. Elevated concentrations of Cu, Zn, and Ni were found in summer, while elevated contents of Pb, Cr, and Co were found in autumn, autumn, and spring, respectively. Trace elements in the western part of Chaohu Lake (especially in the northwestern part) were higher than those in the middle and eastern parts for autumn, winter, and summer. Significant positive correlations were found among Cu, Pb, Zn, Cr, and Ni in the surface water, suggesting that these elements may derive from similar sources. The values of both the single factor pollution index and integrated pollution index of the selected elements in the surface water were far less than 1, suggesting that the environmental impacts could be regarded as negligible. The integrated pollution indices in the western part of the lake were higher than those of the middle and eastern parts on a one-year timescale, and the integrated pollution indices in each lake area followed a pattern of wet season (summer) > normal season (spring and autumn) > dry season (winter).
RESUMEN
BACKGROUND: Muscles converge or intertweave around the perioral area, and this can be treated with sequential therapy in infants with cleft lip and palate (CLP). The force of perioral muscles has a great influence on maxillary development and morphology. Perioral force in infants with CLP has not been well studied, and accurate and reliable measurement of perioral force in infants remains a challenge. This study aimed to investigate a new way to accurately and reliably measure perioral force in infants with unilateral CLP (UCLP) and explore the change before and after cheiloplasty. STUDY DESIGN: A perioral force measurement system was developed and applied to measure perioral force at labial frenum area and the commissures on both the normal and the cleft sides of four infants with UCLP before and after cheiloplasty. The results were analyzed using the SPSS 19.0 software. RESULTS: The perioral force measurement system appears to produce valid results in infants with UCLP. Before cheiloplasty, the perioral force of labial frenum area was 1.79 ± 0.94 g/cm2 and that of commissure on the normal and cleft sides was 5.41 ± 1.01 g/cm2 and 3.12 ± 1.55 g/cm2, respectively (P < 0.05). After cheiloplasty, perioral force of labial frenum area was 12.73 ± 3.51 g/cm2 and that of commissure on the normal and cleft sides was 7.64 ± 1.64 g/cm2 and 7.27 ± 1.89 g/cm2, respectively (P > 0.05).