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1.
Front Immunol ; 15: 1463078, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39445018

RESUMO

Object: Osteosarcoma is a malignant tumor originating from the bones, commonly found in children and adolescents, especially in rapidly growing bone areas such as the knees and upper arms. In this study, we aim to delineate the evolution and convergence of research themes in osteosarcoma metabolomics over the past decade, identify major contributors, and forecast emerging trends that could direct future research efforts. Method: The bibliometric method has been applied to systematically analyze the literature in the field of osteosarcoma metabolomics. The relevant literatures were collected from the Web of Science Core Collection, spanning from January 1, 2014, to December 31, 2023. Tools such as CiteSpace, Bibliometrix, and VOSviewer were used for the visual analysis of the collected literatures. The focused information includes institutions, journals, countries, authors, keywords, and citations. Result: Various aspects in the field of osteosarcoma metabolism were analyzed. Shanghai Jiao Tong University has published the most papers in the past ten years, followed by Central South University and Zhejiang University. Among the sources, the international journal of molecular sciences publishes the most articles, and oncotarget is the journal with the highest H index. According to Bradford's law, there are 34 core journals identified. A total of 5501 authors participated in the creation of papers in this field. The distribution of authors follows Lotka`s Law, and 85.3% of authors have only one article. 46% of the corresponding authors are from China, but most of these corresponding authors are not good at international cooperation. China also has the largest number of publications, followed by the United States. It can be confirmed that China dominates this field. Among the keywords, "expression" is the keyword that has received the most attention in the past ten years. All keywords can be divided into 9 clusters. Based on the explosive words and hot topics each year, we speculate that future research will focus on the tumor microenvironment, molecular mechanisms and autophagy, targeted therapies and inhibitors. Conclusion: In summary, this study comprehensively analyzed the current state of research in the field of osteosarcoma metabolism through bibliometric methods. The findings revealed the development trends and research hotspots in this field, which may provide valuable references for future research directions.


Assuntos
Bibliometria , Neoplasias Ósseas , Metabolômica , Osteossarcoma , Osteossarcoma/metabolismo , Humanos , Metabolômica/métodos , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Pesquisa Biomédica/tendências
2.
Am J Case Rep ; 25: e945262, 2024 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-39420538

RESUMO

BACKGROUND The extraction of impacted supernumerary teeth requires precision and accuracy to mitigate iatrogenic damage to crucial anatomical structures during dental surgical procedures, thereby enhancing postoperative healing outcomes. Dynamic navigation systems (DNS) have been applied in dentistry in maxillofacial fractures, orthognathic surgery, root canal treatment, and endodontic surgery. CASE REPORT A 22-year-old female patient visited our department to assess and manage unerupted third molars. An initial cone beam computed tomography (CBCT) scan was obtained. Radiographic and clinical examinations showed the presence of a supernumerary tooth impacted on the lingual side between the root of the lower second premolar and the lower first molar and bilateral lower impacted third molars. The patient agreed to removal of these teeth. To perform the treatment planning of this case and to guide the surgeon intraoperatively, a dynamic surgical navigation system was recommended for surgical extraction of a supernumerary tooth and the impacted third molars. CONCLUSIONS The dynamic navigation system coupled with a high-speed contra-angle handpiece for the extraction of supernumerary teeth is a personalized, digitally-driven, precise, minimally invasive, and efficient treatment approach. In this case, the DNS and the high-speed contra-angle handpiece were seamlessly integrated to facilitate visualization of the surgical procedure, thereby safeguarding of surrounding vital anatomical structures while enhancing patient comfort.


Assuntos
Mandíbula , Extração Dentária , Dente Supranumerário , Humanos , Feminino , Dente Supranumerário/cirurgia , Dente Supranumerário/diagnóstico por imagem , Adulto Jovem , Mandíbula/cirurgia , Mandíbula/diagnóstico por imagem , Tomografia Computadorizada de Feixe Cônico , Dente Impactado/cirurgia , Dente Impactado/diagnóstico por imagem , Cirurgia Assistida por Computador , Equipamentos Odontológicos de Alta Rotação , Dente Serotino/cirurgia
3.
Front Cell Infect Microbiol ; 14: 1371371, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38524178

RESUMO

Purpose: Human gut microbiota has been shown to be significantly associated with various inflammatory diseases. Therefore, this study aimed to develop an excellent auxiliary tool for the diagnosis of juvenile idiopathic arthritis (JIA) based on fecal microbial biomarkers. Method: The fecal metagenomic sequencing data associated with JIA were extracted from NCBI, and the sequencing data were transformed into the relative abundance of microorganisms by professional data cleaning (KneadData, Trimmomatic and Bowtie2) and comparison software (Kraken2 and Bracken). After that, the fecal microbes with high abundance were extracted for subsequent analysis. The extracted fecal microbes were further screened by least absolute shrinkage and selection operator (LASSO) regression, and the selected fecal microbe biomarkers were used for model training. In this study, we constructed six different machine learning (ML) models, and then selected the best model for constructing a JIA diagnostic tool by comparing the performance of the models based on a combined consideration of area under receiver operating characteristic curve (AUC), accuracy, specificity, F1 score, calibration curves and clinical decision curves. In addition, to further explain the model, Permutation Importance analysis and Shapley Additive Explanations (SHAP) were performed to understand the contribution of each biomarker in the prediction process. Result: A total of 231 individuals were included in this study, including 203 JIA patients and Non-JIA individuals. In the analysis of diversity at the genus level, the alpha diversity represented by Shannon value was not significantly different between the two groups, while the belt diversity was slightly different. After selection by LASSO regression, 10 fecal microbe biomarkers were selected for model training. By comparing six different models, the XGB model showed the best performance, which average AUC, accuracy and F1 score were 0.976, 0.914 and 0.952, respectively, thus being used to construct the final JIA diagnosis model. Conclusion: A JIA diagnosis model based on XGB algorithm was constructed with excellent performance, which may assist physicians in early detection of JIA patients and improve the prognosis of JIA patients.


Assuntos
Artrite Juvenil , Microbiota , Humanos , Artrite Juvenil/diagnóstico , Artrite Juvenil/genética , Biomarcadores , Curva ROC , Aprendizado de Máquina
5.
Sci Rep ; 14(1): 5245, 2024 03 04.
Artigo em Inglês | MEDLINE | ID: mdl-38438569

RESUMO

Osteoporosis is a major public health concern that significantly increases the risk of fractures. The aim of this study was to develop a Machine Learning based predictive model to screen individuals at high risk of osteoporosis based on chronic disease data, thus facilitating early detection and personalized management. A total of 10,000 complete patient records of primary healthcare data in the German Disease Analyzer database (IMS HEALTH) were included, of which 1293 diagnosed with osteoporosis and 8707 without the condition. The demographic characteristics and chronic disease data, including age, gender, lipid disorder, cancer, COPD, hypertension, heart failure, CHD, diabetes, chronic kidney disease, and stroke were collected from electronic health records. Ten different machine learning algorithms were employed to construct the predictive mode. The performance of the model was further validated and the relative importance of features in the model was analyzed. Out of the ten machine learning algorithms, the Stacker model based on Logistic Regression, AdaBoost Classifier, and Gradient Boosting Classifier demonstrated superior performance. The Stacker model demonstrated excellent performance through ten-fold cross-validation on the training set and ROC curve analysis on the test set. The confusion matrix, lift curve and calibration curves indicated that the Stacker model had optimal clinical utility. Further analysis on feature importance highlighted age, gender, lipid metabolism disorders, cancer, and COPD as the top five influential variables. In this study, a predictive model for osteoporosis based on chronic disease data was developed using machine learning. The model shows great potential in early detection and risk stratification of osteoporosis, ultimately facilitating personalized prevention and management strategies.


Assuntos
Neoplasias , Osteoporose , Doença Pulmonar Obstrutiva Crônica , Humanos , Osteoporose/diagnóstico , Osteoporose/epidemiologia , Doença Crônica , Aprendizado de Máquina , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/epidemiologia
6.
Inflamm Res ; 73(5): 693-705, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38150024

RESUMO

BACKGROUND: The aim of this study was to investigate the impact of Porphyromonas gingivalis (P. gingivalis) on the progression of oral squamous cell carcinoma (OSCC) through neutrophil extracellular traps (NETs) in the tumor immune microenvironment. METHODS: The expression of NETs-related markers was identified through immunohistochemistry, immunofluorescence, and Western blotting in different clinical stages of OSCC samples. The relationship between NETs-related markers and clinicopathological characteristics in 180 samples was analyzed using immunohistochemistry data. Furthermore, the ability to predict the prognosis of OSCC patients was determined by ROC curve analysis and survival analysis. The effect of P. gingivalis on the release of NETs was identified through immunofluorescence and immunohistochemistry, both in vitro and in vivo. CAL27 and SCC25 cell lines were subjected to NETs stimulation to elucidate the influence of NETs on various cellular processes, including cell proliferation, migration, invasion, and metastasis in vitro. Furthermore, the impact of NETs on the growth and metastatic potential of OSCC was assessed using in vivo models involving tumor-bearing mice and tumor metastasis mouse models. RESULTS: Immunochemistry analysis revealed a significant correlation between the NETs-related markers and clinical stage, living status as well as TN stage. P. gingivalis has demonstrated its ability to effectively induce the release of NETs both in vivo and in vitro. NETs have the potential to facilitate cell migration, invasion, and colony formation. Moreover, in vivo experiments have demonstrated that NETs play a pivotal role in promoting tumor metastasis. CONCLUSION: High expression of NETs-related markers demonstrates a strong correlation with the progression of OSCC. Inhibition of the NETs release process stimulated by P. gingivalis and targeted NETs could potentially open up a novel avenue in the field of immunotherapy for patients afflicted with OSCC.


Assuntos
Carcinoma de Células Escamosas , Armadilhas Extracelulares , Neoplasias Bucais , Porphyromonas gingivalis , Microambiente Tumoral , Porphyromonas gingivalis/imunologia , Humanos , Armadilhas Extracelulares/imunologia , Armadilhas Extracelulares/metabolismo , Microambiente Tumoral/imunologia , Animais , Neoplasias Bucais/imunologia , Neoplasias Bucais/patologia , Neoplasias Bucais/microbiologia , Linhagem Celular Tumoral , Feminino , Masculino , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Pessoa de Meia-Idade , Camundongos , Progressão da Doença , Camundongos Endogâmicos BALB C , Proliferação de Células , Movimento Celular , Camundongos Nus , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Neutrófilos/imunologia , Idoso
7.
J Cancer ; 14(9): 1660-1672, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37325056

RESUMO

Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common malignancy of the head and neck. However, the molecular mechanisms governing the development of HNSCC have not been fully elucidated. Materials and Methods: Differentially expressed genes (DEGs) were screened out from The Cancer Genome Atlas (TCGA) and GSE23036 datasets. Weighted gene coexpression network analysis (WGCNA) was used to reveal the correlations among genes and to search for significantly correlated gene modules. The expression levels of genes in HNSCC and normal samples according to antibody-based detected methods was assessed by utilizing the Human Protein Atlas (HPA). The impact of the selected hub genes on the prognosis of HNSCC patients was assessed by analysing immunohistochemistry (IHC) and immunofluorescence (IF) expression levels and clinical data. Results: Twenty-four genes positively correlated with tumour status and 15 genes negatively correlated with tumour status were screened out by WGCNA. PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC and were finally screened out and verified by GEPIA and HPA database analysis. Immunohistochemistry of samples collected from 175 patients with HNSCC and subsequent statistical analysis also showed that PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC, and the levels of these two factors were positively correlated. The expression and co-localization of PLAU and LAMC2 in HNSCC tissues were confirmed by double immunofluorescence labeling. Conclusions: There was a positive correlation between PLAU and LAMC2 expression in HNSCC samples, and PLAU and LAMC2 might be independent prognostic biomarkers for HNSCC.

8.
Shanghai Kou Qiang Yi Xue ; 28(2): 154-157, 2019.
Artigo em Chinês | MEDLINE | ID: mdl-31384900

RESUMO

To explore the validity of 3D printing technique in the treatment of unilateral comminuted zygomatic bone fracture. METHODS: Twenty-one patients with unilateral comminuted zygomatic bone fracture were included in the present study, which were treated from hospital January 2014 to April 2017. All patients underwent CT scan and the data were imported in Mimics 10.01 software. The zygomatic bone of healthy side was mirrored to the fracture side to rebuild a "perfect" reduction model. Bone fixation plates were pre-modeled on the model printed by a 3D printing machine and used for bone reduction and fixation during operation. Three dimensional measurements were performed to evaluate the validity of 3D printing based on pre- and post-operative three dimensional CT model. SPSS25.0 software package was used to perform paired t test on the measured data. RESULTS: No significant difference were observed between postoperative CT model and preoperative "perfect" reduction model. All patients were satisfied with their facial appearance. CONCLUSIONS: 3D printing technique is helpful to improve the accuracy of reduction of unilateral comminuted zygomatic bone fracture via preoperative pre-modeling.


Assuntos
Fraturas Cominutivas , Impressão Tridimensional , Fraturas Zigomáticas , Placas Ósseas , Fixação Interna de Fraturas , Fraturas Cominutivas/terapia , Humanos , Fraturas Zigomáticas/terapia
9.
Mol Med Rep ; 18(1): 684-694, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29767244

RESUMO

The use of propranolol for the treatment of infantile hemangioma (IH) has been widely investigated in recent years. However, the underlying therapeutic mechanism of propranolol for the treatment of IH remains poorly understood. The aim of the present study was to investigate the expression of proteins regulated by cellular tumor antigen p53 (p53) in associated apoptosis pathways in IH endothelial cells (HemECs) treated with propranolol. Furthermore, the present study aimed to investigate the exact apoptotic pathway underlying the therapeutic effect of propranolol against IH. In the present study, HemECs were subcultured and investigated using an inverted phase contrast microscope, immunocytochemical staining and a scanning electron microscope (SEM). Experimental groups and blank control groups were prepared. All groups were subjected to drug treatment. A high p53 expression model of HemECs was successfully established via transfection, and a low p53 expression model of HemECs was established using pifithrin­α. The apoptosis rate of each group was determined using Annexin V­fluorescein isothiocyanate/propidium iodide double staining and flow cytometry. The expression levels of downstream proteins regulated by p53 [tumour necrosis factor receptor superfamily member 6 (FAS), p53­induced death domain­containing protein (PIDD), death receptor 5 (DR5), BH3­interacting domain death agonist (BID), apoptosis regulator BAX (BAX), p53 unregulated modulator of apoptosis (PUMA), phosphatidylinositol­glycan biosynthesis class S protein (PIGS), and insulin­like growth factor­binding protein 3 (IGF­BP3)] were revealed in the experimental and control groups via western blotting. Microscopic observation revealed the growth of an adherent monolayer of cells, which were closely packed and exhibited contact inhibition. Immunocytochemical staining demonstrated increased expression of clotting factor VIII. SEM analysis revealed presence of Weibel­Palade bodies. The results of the analyses verified that the cultured cells were HemECs. The staining of the samples resulted in a significantly increased rate of apoptosis in experimental groups compared with the blank control group. This result suggested that there is an association between p53 expression and the rate of apoptosis of propranolol­treated HemECs. The results of the western blot analysis demonstrated an upregulation of BAX expression and a downregulation of IGF­BP3 expression in the HemECs treated with propranolol. There were no significant differences in the expression levels of FAS, DR5, PIDD, BID, PUMA and PIGS between experimental and control groups. This result suggests that p53 has an important role in HemEC apoptosis. The results of the present study additionally suggest that the propranolol­induced HemEC apoptosis pathway is a mitochondrial apoptosis pathway and is regulated by p53­BAX signaling.


Assuntos
Apoptose/efeitos dos fármacos , Hemangioma/tratamento farmacológico , Hemangioma/metabolismo , Propranolol/efeitos adversos , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo , Apoptose/genética , Células Endoteliais , Feminino , Hemangioma/genética , Hemangioma/patologia , Humanos , Masculino , Mitocôndrias/genética , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Propranolol/farmacologia , Transdução de Sinais/genética , Proteína Supressora de Tumor p53/genética , Corpos de Weibel-Palade/genética , Corpos de Weibel-Palade/metabolismo , Corpos de Weibel-Palade/patologia , Proteína X Associada a bcl-2/genética
10.
Mol Med Rep ; 10(1): 301-5, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24789513

RESUMO

Pingyangmycin (also known as Bleomycin A5) is produced by Streptomyces verticillus var. pingyangensis n.sp., and has anti­tumor activities against a variety of tumor cells. The aim of the present study was to determine the molecular mechanism(s) underlying the therapeutic effects of pingyangmycin against infantile hemangiomas. Human hemangioma­derived endothelial cells (HemECs) were treated with pingyangmycin at varying concentrations (100, 200 or 300 µg/ml), and the morphological changes and apoptosis levels were assessed. The gene expression changes were determined by cDNA microarray technology. Transmission electron microscopy examination revealed that the pingyangmycin­treated HemECs exhibited typical apoptotic characteristics, including chromatin condensation and the formation of apoptotic bodies. Annexin­V staining demonstrated that pingyangmycin caused a significant and dose­dependent induction of apoptosis in the HemECs. In the pingyangmycin­treated HemECs, 4,752 genes demonstrated at least 2­fold expression changes at the mRNA level. Quantitative polymerase chain reaction confirmed that pingyangmycin significantly upregulated the expression of p53, p53­induced protein with death domain, Bax, p53 upregulated modulator of apoptosis and p53 inducible gene 3, and downregulated the expression of murine double minute 2. The data demonstrated that the pro­apoptotic activity of pingyangmycin against infantile hemangiomas involves p53 pathway activation.


Assuntos
Antibióticos Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Bleomicina/análogos & derivados , Células Endoteliais/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , Bleomicina/toxicidade , Regulação para Baixo/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Hemangioma/metabolismo , Hemangioma/patologia , Humanos , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , RNA Mensageiro/metabolismo , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética , Regulação para Cima/efeitos dos fármacos , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
11.
Exp Ther Med ; 6(2): 574-578, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24137229

RESUMO

Propranolol, a non-selective ß-blocker, is emerging as an effective treatment for complicated hemangiomas. The aim of this study was to investigate the molecular mechanism(s) underlying the therapeutic effects of propranolol against hemangiomas, using primary infantile hemangioma endothelial cells (IHECs). IHECs were treated with various concentrations of propranolol and morphological changes and apoptosis were assessed. Changes in the expression levels of apoptosis-related genes were examined. Annexin-V staining revealed that propranolol at 40, 50 and 60 µg/ml caused a concentration-dependent increase in the apoptosis of IHECs. Morphological analyses revealed that exposure to 50 µg/ml propranolol resulted in typical apoptotic changes, including shrinkage, the formation of apoptotic bodies and retention of plasma membrane integrity. Gene expression analyses revealed that propranolol treatment led to a marked increase in the expression of caspase-8, cytochrome c, apoptosis-inducing factor, caspase-3 and poly (ADP-ribose) polymerase 1, as well as a concomitant reduction in lamin B1 expression. Our data collectively demonstrate that propranolol induces apoptosis of IHECs through activation of the intrinsic and extrinsic apoptotic pathways, which represents an important mechanism for its therapeutic effects against infantile hemangiomas.

12.
J Biochem Mol Toxicol ; 26(9): 374-80, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22987598

RESUMO

Hemangioma is the most common benign tumor of infancy. The aim of this study is to evaluate the biological effects of sodium morrhuate (SM) and its liposomal formulation on infantile hemangioma endothelial cells (IHECs). Morphological analysis revealed that exposure to liposomal sodium morrhuate (LSM) preferentially caused apoptotic death in IHECs, manifested as shrunken configuration and formation of apoptotic bodies. In contrast, necrotic death was prominent in IHECs treated with an equal concentration of SM. By means of proteomic analysis and confirmation experiments, we revealed that the apoptosis-inducing effects of LSM were associated with an upregulation of a set of genes involved in mitochondrial death pathway, including apoptosis-inducing factor, cytochrome c1, caspase-8, and lamin B1. In conclusion, our data highlight the proapoptotic activity of LSM in IHECs through the mitochondrial apoptotic pathway and may provide a promising avenue to treat hemangiomas of infancy.


Assuntos
Antineoplásicos/farmacologia , Células Endoteliais/efeitos dos fármacos , Hemangioma/metabolismo , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Proteoma/metabolismo , Morruato de Sódio/farmacologia , Apoptose/efeitos dos fármacos , Fator de Indução de Apoptose/genética , Fator de Indução de Apoptose/metabolismo , Caspase 8/genética , Caspase 8/metabolismo , Forma Celular , Citocromos c1/genética , Citocromos c1/metabolismo , Composição de Medicamentos , Expressão Gênica/efeitos dos fármacos , Hemangioma/tratamento farmacológico , Hemangioma/patologia , Humanos , Lactente , Lamina Tipo B/genética , Lamina Tipo B/metabolismo , Lipossomos , Proteínas Mitocondriais/genética , Proteoma/genética , Proteômica , Células Tumorais Cultivadas
13.
Artif Organs ; 34(2): 161-6, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20420593

RESUMO

To develop a cartilage-like tissue with hybrid scaffolds of demineralized bone matrix gelatin (BMG) and fibrin, rabbit chondrocytes were cultured on hybrid fibrin/BMG scaffolds in vitro. BMG scaffolds were carefully soaked in a chondrocyte-fibrin suspension, which was polymerized by submerging the constructs into thrombin-calcium chloride solution. Engineered cartilage-like tissue grown on the scaffolds was characterized by histology, immunolocalization, scanning electron microscopy, biochemical assays, and analysis of gene expression at different time points of the in vitro culture. The presence of proteoglycan in the fibrin/BMG hybrid constructs was confirmed by positive toluidine blue and alcian blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene-encoded cartilage-specific markers, collagen type II, and aggrecan core protein. The glycosaminoglycan production and hydroxyproline content of tissue grown on the fibrin/BMG hybrid scaffolds were higher than that of the BMG group. In conclusion, the fibrin/BMG hybrid scaffolds may serve as a potential cell delivery vehicle and a structural basis for cartilage tissue engineering.


Assuntos
Matriz Óssea/metabolismo , Cartilagem/metabolismo , Engenharia Tecidual , Alicerces Teciduais , Animais , Materiais Biocompatíveis/metabolismo , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Fibrina/metabolismo , Adesivo Tecidual de Fibrina/metabolismo , Gelatina/metabolismo , Imuno-Histoquímica , Microscopia Eletrônica de Varredura , Coelhos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Nan Fang Yi Ke Da Xue Xue Bao ; 29(9): 1766-9, 2009 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-19778785

RESUMO

OBJECTIVE: To study the effect of RNA interference (RNAi)-mediated aggrecanase-1 gene silencing on extracellular matrix metabolism of cultured rat costochondral chondrocytes. METHODS: Rat costochondral chondrocyte monolayers were obtained by microdissection and digestion. The growth and morphological changes of the chondrocytes were observed after RNAi of aggrecanase-1 gene. The mRNA expression of aggrecanase-1 was detected by RT-PCR method, and aggrecan content was determined by Western blotting. RESULTS: The specific inhibition of aggrecanase-1 by RNAi produced no adverse effect on the morphology and growth of the chondrocytes. The mRNA of aggrecanase-1 decreased and aggrecan content increased significantly after transfection of the chondrocytes. CONCLUSION: Inhibition of aggrecanase-1 decreases aggrecan degradation in cultured rat chondrocytes. RNAi technique can be a useful means for studying extracellular matrix metabolism in the cartilage.


Assuntos
Proteínas ADAM/genética , Condrócitos/citologia , Matriz Extracelular/metabolismo , Pró-Colágeno N-Endopeptidase/genética , Interferência de RNA , Proteínas ADAM/metabolismo , Proteína ADAMTS4 , Agrecanas/metabolismo , Animais , Células Cultivadas , Condrócitos/metabolismo , Feminino , Pró-Colágeno N-Endopeptidase/metabolismo , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Transfecção
15.
Acta Pharmacol Sin ; 29(10): 1215-26, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18817627

RESUMO

AIM: Failure of transplanted cartilage or allogenic chondrocytes is attributed mainly to immunological rejection and cartilage degradation. A major feature is the loss of aggrecan from the cartilage matrix, primarily due to the action of the specific proteinases aggrecanase-1 and aggrecanase-2. The aim of this in vitro study was to determine whether the specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi would mitigate aggrecan loss from cultured chondrocytes. METHODS: Expression plasmid vectors of shRNA targeting aggrecanase-1 and aggrecanase-2 were constructed and transfected into cultured rattus costochondral chondrocytes. The transfected cells were induced with interleukin-1beta (IL-1beta). Gene mRNA levels were analyzed by RT-PCR. Aggrecan and collagen II content were measured by immunohistochemistry and Western blotting. RESULTS: As the chondrocytes underwent dedifferentiation, aggrecanase-1 increased significantly. The specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi had no negative effect on the morphology and growth velocity of the chondrocytes. The mRNA of aggrecanase-1 and aggrecanase-2 decreased significantly. The alpha-2-macroglobulin expression level was increased by the shRNA specific for aggrecanase-1. Other genes of the chondrocytic extracellular matrix were not affected. RNAi significantly increased the aggrecan and collagen II content of chondrocytes treated with IL-1beta. CONCLUSION: The results suggest that inhibition of aggrecanase-1 and aggrecanase-2 by RNAi can mitigate aggrecan degradation, without interfering with chondrocytic gene phenotype recovery. RNAi technology can be a useful tool for studying degenerative processes in cartilage.


Assuntos
Proteínas ADAM/antagonistas & inibidores , Condrócitos/efeitos dos fármacos , Condrócitos/enzimologia , Pró-Colágeno N-Endopeptidase/antagonistas & inibidores , Interferência de RNA/fisiologia , Proteína ADAMTS4 , Proteína ADAMTS5 , Agrecanas/metabolismo , Animais , Colágeno Tipo II/metabolismo , Interleucina-1beta/biossíntese , Interleucina-1beta/genética , Plasmídeos/genética , Ratos , Ratos Sprague-Dawley , Transfecção , alfa-Macroglobulinas/biossíntese
16.
Shanghai Kou Qiang Yi Xue ; 17(2): 143-50, 2008 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-18470417

RESUMO

PURPOSE: To explore the method of preparing immunolipo-sodium morrhuate and evaluate its effect on human hemangioma endothelial cells in vitro. METHODS: Using SPDP((N-Succinimidyl-3-(2-pyridyldithio)) propionate) as cross-linker, anti-VEGFR2/KDR monoclone antibody was combined to the liposome surface to prepare immunolipo-sodium morrhuate by extruding method, and then its effect on human hemangioma endothelial cells in vitro was observed by laser scanning confocal microscope, inverted microscope, Gimsa staining, transmission electron microscope, MTT and flow cytometry. RESULTS: The average diameter of the immunoliposome was 122.9 nm, which had a very good stability when compared with normal liposome, it had stronger and faster combining ability, its potential to induce apoptosis was much more prominent, and its toxic effect on human hemangioma endothelial cells was gentle, which was similar to normal liposome. CONCLUSION: We have prepared immunolipo-sodium morrhuate successfully, which has very good specific initiative targetting ability in vitro and can induce pervasive apoptosis of human hemangioma endothelial cells.


Assuntos
Apoptose/efeitos dos fármacos , Hemangioma/patologia , Soluções Esclerosantes/farmacologia , Morruato de Sódio/farmacologia , Células Endoteliais , Humanos , Técnicas In Vitro , Lipossomos
17.
Zhonghua Zheng Xing Wai Ke Za Zhi ; 24(1): 50-3, 2008 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-18437986

RESUMO

OBJECTIVE: To observe the effect of aloesin, tea polyphenols, arbutin on melanocytes in the pigmented skin equivalent model. METHODS: First, we constructed the pigmented skin equivalent model in vitro. And then we detected the effect of aloesin, tea polyphenols and arbutin on the cells' shape, tyrosinase activity and formation of melanin in the constructed pigmented skin equivalent. RESULTS: Three depigmenting agents showed an inhibition effect on the tyrosinase activity of melanocytes and reduced significantly melanin content in the pigmented skin equivalent model, in which the tea polyphenols had the strongest effect, and then was the aloesin. But the tea polyphenols showed the strongest toxicity, while the aloesin and arbutin had a much lower toxicity. CONCLUSIONS: All the three depigmenting agents showed a concentration dependent suppression effect on the tyrosinase activity and formation of melanin, in which the tea polyphenols was the strongest effect( P <0.05). Aoesin has a good suppression effect on the tyrosinase activity and formation of melanin, but has a much lower toxicity, which could be used as a safe depigmenting agent.


Assuntos
Arbutina/farmacologia , Cromonas/farmacologia , Flavonoides/farmacologia , Glucosídeos/farmacologia , Melanócitos/efeitos dos fármacos , Fenóis/farmacologia , Células Cultivadas , Prepúcio do Pênis/citologia , Humanos , Masculino , Melaninas/biossíntese , Pigmentação , Polifenóis , Pele/efeitos dos fármacos , Envelhecimento da Pele/efeitos dos fármacos
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 27(11): 1670-3, 2007 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-18024286

RESUMO

OBJECTIVE: To construct an in vitro equivalent of the pigmented skin using tissue engineering methods. METHODS: Surgically removed foreskins was used as the source of keratinocytes and melanocytes harvested by routine tissue digestion. The fibroblasts were enriched by tissue block culture and seeded into the scaffold constructed using mouse tail collagens to construct the pigmented skin equivalent model. The general structure and the melanocyte distribution and growth status in this model were observed with HE staining and Fontana Masson staining. The ultrastructure of the constructed pigmented skin equivalent was observed by transmission electron microscope. RESULTS AND CONCLUSION: The pigmented skin equivalent model was structurally intact, and allowed optimal cell growth. Fontana Masson staining identified in the basal layer numerous melanocytes in normal growth, and the constructed model was structurally similar to normal skin tissue, suggesting successful construction of the pigmented skin equivalent model.


Assuntos
Pigmentação da Pele , Pele Artificial , Engenharia Tecidual/métodos , Animais , Prepúcio do Pênis/citologia , Humanos , Queratinócitos/citologia , Masculino , Melanócitos/citologia , Camundongos
19.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 42(3): 188-9, 2007 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-17565835

RESUMO

OBJECTIVE: To evaluate the effect of lipo-sodium morrhuate on ECV-304 cell line. METHODS: The effect lipo-sodium morrhuate was evaluated by toxicology trial (MTT), electron microscope, DNA electrophoresis and flow cytometer. RESULTS: The toxicology results showed, that the number of vital cells in lipo-sodium morrhuate group decreased slowly. The electron microscope exhibited apoptosis in the lipo-sodium morrhuate group. And there were typical DNA ladder in DNA electrophoresis and typical apoptosis peak in flow cytometer. The apoptosis rate was 22.23%. CONCLUSIONS: Unlike the normal preparation of sodium morrhuate, lipo-sodium morrhuate could induce apoptosis of ECV-304 cell line.


Assuntos
Células Endoteliais/patologia , Morruato de Sódio/farmacologia , Veias Umbilicais/citologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Humanos , Lipossomos
20.
Zhonghua Zheng Xing Wai Ke Za Zhi ; 23(1): 59-61, 2007 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-17393698

RESUMO

OBJECTIVE: The aim of this study was to observe the human hair follicle apoptosis status affected by fluorine and the antagonism effect by selenium in vitro. METHODS: The single hair follicles were separated and cultured, then they were added in different concentrations of sodium fluoride and sodium selenite. Chosen the appropriate concentrations, they were divided into 7 groups. The TUNEL was used to investigate the apoptotic cells of different parts. The morphous of hair follicles was observed consecutively and electron microscope was used. RESULTS: We found that in 1 mmol/L and 10 mmol/L sodium fluoride groups, when the human hair follicles in vitro were cultured on the 5th day, the apoptotic cells of outer root sheath (ORS), dermal sheath and hair papilla, hair bulb were obviously increased. But 0.01 mmol/L sodium selenite weakened the toxicity of 1 mmol/L sodium fluoride at the outer root sheath and hair bulb (P < 0.05). CONCLUSIONS: Different concentrations of sodium fluoride had different effect on the growth of human hair follicle in vitro which were cultured on 5th day. Sodium fluoride of certain concentration could accelerate the apoptosis of human hair follicle in vitro. Sodium selenite of certain concentration could act antagonism to the toxicity of sodium fluoride.


Assuntos
Apoptose/efeitos dos fármacos , Folículo Piloso/efeitos dos fármacos , Fluoreto de Sódio/farmacologia , Adolescente , Adulto , Humanos , Pessoa de Meia-Idade , Selenito de Sódio/farmacologia , Técnicas de Cultura de Tecidos , Adulto Jovem
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