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1.
J Bone Miner Metab ; 40(2): 240-250, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35066669

RESUMEN

INTRODUCTION: The diagnosis and treatment of osteoporosis, a frequent age-related metabolic bone disorder, remain incomprehensive and challenging. The potential regulatory role of lncRNA XIST and sphingosine kinase 1 (SPHK1) pathway need experimental investigations. MATERIALS AND METHODS: RAW264.7 cells and BMMs were obtained for in vitro studies and 30 ng/mL RANKL was implemented for induction of osteoclast differentiation. The suppressing of lncRNA XIST, SPHK1 and fused in sarcoma (FUS) was achieved using small hairpin RNA, while overexpression of XIST and FUS was constructed by pcDNA3.1 vector system. Tartrate-resistant acid phosphatase (TRAP) staining was used for observation of formation of osteoclasts. RNA-pulldown analysis and RNA binding protein immunoprecipitation (RIP) was implemented for measuring mRNA and protein interactions. RT-qPCR was conducted to determining mRNA expression, whereas ELISA and Western blotting assay was performed for monitoring protein expression. RESULTS: RANKL induced osteoclast differentiation and upregulated expression of osteoclastogenesis-related genes that included NFATc1, CTSK, TRAP and SPHK1 and the level of lncRNA XIST in both RAW264.7 cells and BMMs. However, knockdown of lncRNA XIST or suppressing SPHK1 significantly reserved the effects of RANKL. LncRNA XIST was further demonstrated to be interacted with FUS and increased the stability of SPHK1, indicating its ability in promoting osteoclast differentiation through SPHK1/S1P/ERK signaling pathway. CONCLUSION: LncRNA XIST promoted osteoclast differentiation via interacting with FUS and upregulating SPHK1/S1P/ERK pathway.


Asunto(s)
Resorción Ósea , Osteoclastos , Proproteína Convertasas/metabolismo , ARN Largo no Codificante , Proteína FUS de Unión a ARN/metabolismo , Serina Endopeptidasas/metabolismo , Animales , Resorción Ósea/metabolismo , Catepsina K/metabolismo , Diferenciación Celular , Hematopoyesis , Ratones , Factores de Transcripción NFATC/metabolismo , Osteoclastos/citología , Osteogénesis , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Ligando RANK/metabolismo , Células RAW 264.7 , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Fosfatasa Ácida Tartratorresistente/metabolismo
2.
J Mater Sci Mater Med ; 27(5): 89, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26975746

RESUMEN

To develop adriamycin (ADM)-encapsulated poly(lactic-co-glycolic acid) (PLGA) nanoparticles in a porous nano-hydroxyapatite/collagen scaffold (ADM-PLGA-NHAC). To provide novel strategies for future treatment of osteosarcoma, the properties of the scaffold, including its in vitro extended-release properties, the inhibition effects of ADM-PLGA-NHAC on the osteosarcoma MG63 cells, and its bone repair capacity, were investigated in vivo and in vitro. The PLGA copolymer was utilized as a drug carrier to deliver ADM-PLGA nanoparticles (ADM-PLGA-NP). Porous nano-hydroxyapatite and collagen were used to materials to produce the porous nano-hydroxyapatite/collagen scaffold (NHAC), into which the ADM-PLGA-NP was loaded. The performance of the drug-carrying scaffold was assessed using multiple techniques, including scanning electron microscopy and in vitro extended release. The antineoplastic activities of scaffold extracts on the human osteosarcoma MG63 cell line were evaluated in vitro using the cell counting kit-8 (CCK8) method and live-dead cell staining. The bone repair ability of the scaffold was assessed based on the establishment of a femoral condyle defect model in rabbits. ADM-PLGA-NHAC and NHAC were implanted into the rat muscle bag for immune response experiments. A tumor-bearing nude mice model was created, and the TUNEL and HE staining results were observed under optical microscopy to evaluate the antineoplastic activity and toxic side effects of the scaffold. The composite scaffold demonstrated extraordinary extended-release properties, and its extracts also exhibited significant inhibition of the growth of osteosarcoma MG63 cells. In the bone repair experiment, no significant difference was observed between ADM-PLGA-NHAC and NHAC by itself. In the immune response experiments, ADM-PLGA-NHAC exhibited remarkable biocompatibility. The in vivo antitumor experiment revealed that the implantation of ADM-PLGA-NHAC in the tumor resulted in a improved antineoplastic effect and fewer adverse side effects than direct intraperitoneal injection of ADM. The ADM-PLGA-NHAC developed in this study exhibited excellent extended-release drug properties, bone repairing and antineoplastic efficacy, which make it a promising osteoconductivity material with the capability to inhibit osteosarcoma.


Asunto(s)
Neoplasias Óseas/tratamiento farmacológico , Colágeno/química , Doxorrubicina/química , Durapatita/química , Ácido Láctico/química , Ácido Poliglicólico/química , Andamios del Tejido/química , Animales , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacología , Línea Celular Tumoral , Doxorrubicina/farmacología , Femenino , Humanos , Ácido Láctico/farmacología , Masculino , Ratones , Ratones Desnudos , Nanoestructuras , Osteosarcoma , Ácido Poliglicólico/farmacología , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Conejos , Ratas , Ratas Sprague-Dawley
3.
Orthop Surg ; 14(9): 2256-2264, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35979964

RESUMEN

OBJECTIVE: 3D reconstruction of lumbar intervertebral foramen (LIVF) has been beneficial in evaluating surgical trajectory. Still, the current methods of reconstructing the 3D LIVF model are mainly based on manual segmentation, which is laborious and time-consuming. This study aims to explore the feasibility of automatically segmenting lumbar spinal structures and increasing the speed and accuracy of 3D lumbar intervertebral foramen (LIVF) reconstruction on magnetic resonance image (MRI) at the L4-5 level. METHODS: A total of 100 participants (mean age: 42.2 ± 14.0 years; 52 males and 48 females; mean body mass index, 22.7 ± 3.2 kg/m2 ), were enrolled in this prospective study between March and July 2020. All participants were scanned on L4-5 level with a 3T MR unit using 3D T2-weighted sampling perfection with application-optimized contrast with various flip-angle evolutions (SPACE) sequences. The lumbar spine's vertebra bone structures (VBS) and intervertebral discs (IVD) were manually segmented by skilled surgeons according to their anatomical outlines from MRI. Then all manual segmentation were saved and used for training. An automated segmentation method based on a 3D U-shaped architecture network (3D-UNet) was introduced for the automated segmentation of lumbar spinal structures. A number of quantitative metrics, including dice similarity coefficient (DSC), precision, and recall, were used to evaluate the performance of the automated segmentation method on MRI. Wilcoxon signed-rank test was applied to compare morphometric parameters, including foraminal area, height and width of 3D LIVF models between automatic and manual segmentation. The intra-class correlation coefficient was used to assess the test-retest reliability and inter-observer reliability of multiple measurements for these morphometric parameters of 3D LIVF models. RESULTS: The automatic segmentation performance of all spinal structures (VBS and IVD) was found to be 0.918 (healthy levels: 0.922; unhealthy levels: 0.916) for the mean DSC, 0.922 (healthy levels: 0.927; unhealthy levels: 0.920) for the mean precision, and 0.917 (healthy levels: 0.918; unhealthy levels: 0.917) for the mean recall in the test dataset. It took approximately 2.5 s to achieve each automated segmentation, far less than the 240 min for manual segmentation. Furthermore, no significant differences were observed in the foraminal area, height and width of the 3D LIVF models between manual and automatic segmentation images (P > 0.05). CONCLUSION: A method of automated MRI segmentation based on deep learning algorithms was capable of rapidly generating accurate segmentation of spinal structures and can be used to construct 3D LIVF models from MRI at the L4-5 level.


Asunto(s)
Aprendizaje Profundo , Imagenología Tridimensional , Adulto , Femenino , Humanos , Imagenología Tridimensional/métodos , Vértebras Lumbares/diagnóstico por imagen , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Reproducibilidad de los Resultados
4.
J Int Med Res ; 48(4): 300060519894434, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31875742

RESUMEN

Primary Sjögren's syndrome (pSS) is associated with an increased risk of lymphoma, especially non-Hodgkin's lymphoma. The rarest pathological subtype is T-cell lymphoma. We herein report a case of a 52-year-old man with a 17-year history of pSS who was admitted to our hospital with chronic epigastric pain and a positive fecal occult blood test. Colonoscopy revealed multiple colonic ulcers, and histological and immunological studies demonstrated the T-cell origin of this lymphoma. However, the patient rejected all treatments. He developed recurrent intestinal obstruction and infection for 3 years until an intestinal perforation occurred. The right half of the colon was resected and colostomy was performed. However, the patient died of an intestinal fistula and intraperitoneal infection 40 days postoperatively. This case highlights the rarity of the correlation between T-cell lymphoma and pSS.


Asunto(s)
Linfoma no Hodgkin , Linfoma de Células T , Síndrome de Sjögren , Colon , Colostomía , Humanos , Linfoma de Células T/complicaciones , Masculino , Persona de Mediana Edad , Síndrome de Sjögren/complicaciones
5.
Oncotarget ; 9(12): 10483-10496, 2018 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-29535821

RESUMEN

Osteosarcoma(OS) remains a major health concern in childhood and adolescence, although cisplatin is one of the gold standard chemotherapeutic drugs in the treatment of OS, chemoresistant to cisplatin is common. Phosphoinositide 3-kinase (PI3K)-Akt-mammalian target of rapamycin inhibitor (mTOR) pathway and autophagy regulates chemosensitivity incancer cells. In this study, we hypothesized that NVP-BEZ235, a dual inhibitor of PI3K/mTOR, could synergize cisplatin sensitivity in OS. In vitro, NVP-BEZ235 plus cisplatinexerted a synergistic effect on cell proliferation inhibition and apoptosis induction. Cisplatin could activate PI3K-Akt-mTOR pathway activity in early times, whereas, NVP-BEZ235 could inhibit PI3K-Akt -mTOR pathway activity all the times alone or combined with cisplatin. What's more, NVP-BEZ235 could switch function of autophagy induced by cisplatin to synergize cisplatin sensitivity. In vivo, pronounced decrease in tumor cell proliferation and increase in apoptosisin combination-treated mouse xenograft models compared with cisplatin or NVP-BEZ235 treated models. All these results suggest NVP-BEZ235 could synergize cisplatin sensitivity in OS, combination of NVP-BEZ235 with cisplatin could represent a novel therapeutic strategy for treatment of OS.

6.
World J Gastroenterol ; 8(5): 808-14, 2002 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-12378620

RESUMEN

AIM: To elucidate the expression of the apoptosis-associated molecules in human primary hepatocellular carcinoma (HCC) cells, and prepare the monoclonal antibodies (mAb) against the apoptosis-associated antigens of HCC cells. METHODS: Human HCC cell line HCC-9204 cells were induced apoptosis with 60 mL x L(-1) ethanol for 6 h and their morphological changes were observed by transmission electron microscope. The cell DNA fragmentations were detected by Terminal Deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and the cell DNA contents by flow cytometry. Ten mice were immunized with ethanol-induced apoptotic HCC-9204 cells with the method of subtractive immunization, while the other 10 mice used as the control were immunized by the routine procedures. The tail blood of all the mice were prepared after the last immunization, and the produced antibodies were determined by the immunocytochemical ABC staining. The splenic cells of the mice whose tail blood sera-HCC-9204 cells serum reactions were most different between the apoptotic and the non-apoptotic were prepared and fused with the mouse myeloma cell line SP2/0 cells. The positive antibodies were selected by ELISA assay. The fusion rates of hybridoma cells and the producing rates of antibodies were calculated. The fused cells that secreted candidate objective antibody were cloned continually with the of limited dilution method, and then selected and analyzed further by the immunocytochemical ABC staining. The chromosomes of the cloned hybridoma cells that secreted objective mAb and the mAb immunoglobulin (Ig) subtype of the prepared mAb were also determined. The molecular mass of the mAb associated antigen was analyzed by Western blot assay. RESULTS: HCC-9204 cells treated with 60 mL x L(-1) ethanol for 6 h, manifested obvious apoptotic morphological changes, the majority of the cells were TUNEL-positive, and the sub-G1 apoptotic peak was evident. There were 2 mice in the experimental group whose tail blood serum reacted strongly with the apoptotic HCC-9204 cells, but weakly with their non-apoptotic counterparts. In the fusion rates of hybridoma cells as well as the producing rates of the antibody described above, there did not show significant difference between the experimental and the control group, but weakly with non-apoptotic HCC-9204. However, the total producing rate of antibodies in the experimental group was significantly lower compared with the control (P<0.01), and so was the producing rate of the antibodies which reacted strongly with both apoptotic and non-apoptotic HCC-9204 cells(P<0.01). After cloned continually for several times the cell that produce mAb which reacted strongly with the nuclei of ethanol-induced apoptotic HCC-9204 cells, but very weakly with that of non-apoptotic cells was selected out. Chromosome analysis revealed that the selected cell was with the universal characteristics of the monoclonal hybridoma cells which secreted mAb, and the Ig subtype of the prepared mAb was IgG1. The molecular mass of this mAb associated antigen of was about 75 ku. CONCLUSION: Subtractive immunization is a useful method to prepare the mAb against the apoptosis-associated antigens of cells. The expression of some molecules increases to some extent in HCC-9204 cells in the process of apoptosis induced by low-concentration ethanol. The mAb that may be against ethanol-induced apoptosis-associated antigens of HCC cells was successfully prepared and primarily identified.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Antígenos de Neoplasias/inmunología , Apoptosis/inmunología , Carcinoma Hepatocelular , Neoplasias Hepáticas , Antígenos de Neoplasias/química , Fusión Celular , ADN de Neoplasias/análisis , Humanos , Hibridomas , Inmunización , Inmunoglobulina G/inmunología , Microscopía Electrónica , Peso Molecular , Células Tumorales Cultivadas/inmunología , Células Tumorales Cultivadas/ultraestructura
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