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1.
J Clin Ultrasound ; 52(3): 274-283, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38105371

RESUMO

BACKGROUND: Explore the feasibility of using the multimodal ultrasound (US) radiomics technology to diagnose American College of Radiology (ACR) Thyroid Imaging Reporting and Data System (TI-RADS) 4-5 thyroid nodules. METHOD: This study prospectively collected the clinical characteristics, conventional, and US elastography images of 100 patients diagnosed with ACR TI-RADS 4-5 nodules from May 2022 to 2023. Independent risk factors for malignant thyroid nodules were extracted and screened using methods such as the least absolute shrinkage and selection operator (LASSO) logistic regression (LR) model, and a multimodal US radiomics combined diagnostic model was established. Using a multifactorial LR analysis and a Rad-score rating, the predictive performance was validated and evaluated, and the final threshold range was determined to assess the clinical net benefit of the model. RESULTS: In the training set, the US radiomics combined predictive model area under curve (AUC = 0.928) had higher diagnostic performance compared with clinical characteristics (AUC = 0.779), conventional US (AUC = 0.794), and US elastography model (AUC = 0.852). In the validation set, the multimodal US radiomics combined diagnostic model (AUC = 0.829) also had higher diagnostic performance compared with clinical characteristics (AUC = 0.799), conventional US (AUC = 0.802), and US elastography model (AUC = 0.718). CONCLUSION: Multi-modal US radiomics technology can effectively diagnose thyroid nodules of ACR TI-RADS 4-5, and the combination of radiomics signature and conventional US features can further improve the diagnostic performance.


Assuntos
Técnicas de Imagem por Elasticidade , Nódulo da Glândula Tireoide , Humanos , Nódulo da Glândula Tireoide/diagnóstico por imagem , Nódulo da Glândula Tireoide/patologia , Radiômica , Estudos Retrospectivos , Ultrassonografia/métodos , Tecnologia
2.
Front Oncol ; 13: 1060702, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37251934

RESUMO

Artificial intelligence (AI), particularly deep learning (DL) algorithms, has demonstrated remarkable progress in image-recognition tasks, enabling the automatic quantitative assessment of complex medical images with increased accuracy and efficiency. AI is widely used and is becoming increasingly popular in the field of ultrasound. The rising incidence of thyroid cancer and the workload of physicians have driven the need to utilize AI to efficiently process thyroid ultrasound images. Therefore, leveraging AI in thyroid cancer ultrasound screening and diagnosis cannot only help radiologists achieve more accurate and efficient imaging diagnosis but also reduce their workload. In this paper, we aim to present a comprehensive overview of the technical knowledge of AI with a focus on traditional machine learning (ML) algorithms and DL algorithms. We will also discuss their clinical applications in the ultrasound imaging of thyroid diseases, particularly in differentiating between benign and malignant nodules and predicting cervical lymph node metastasis in thyroid cancer. Finally, we will conclude that AI technology holds great promise for improving the accuracy of thyroid disease ultrasound diagnosis and discuss the potential prospects of AI in this field.

3.
Environ Sci Pollut Res Int ; 30(4): 9416-9427, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36050557

RESUMO

In this study, TiO2/MSGM composite material with adsorption-photocatalytic properties was prepared by extracting mesoporous silica glass microsphere (MSGM) from coal gasification fine slag (CGFS) as a novel TiO2 carrier. The results of characterization and properties of the composite showed that MSGM could improve the adsorption capacity and photocatalytic activity of the composite by improving the pore structure of the composite, hindering the growth of TiO2 particles, increasing the phase transition temperature of TiO2, enhancing the dispersion of TiO2 particles. The sample 1:3-TiO2/MSGM-2-500 prepared under the optimized conditions possesses satisfactory morphology characteristics, high adsorption capacity, and photocatalytic activity to rhodamine B (RhB). The synergistic effects of adsorption and photocatalytic significantly increase the total removal rate of RhB. This study not only provides a new direction for high-value-added resource utilization of CGFS but also gives a new kind of low-cost carrier material with adsorption property for TiO2 loading to remove organic dye pollutants.


Assuntos
Carvão Mineral , Dióxido de Silício , Adsorção , Microesferas
4.
Front Oncol ; 12: 990603, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36439514

RESUMO

Objective: This study compared the diagnostic value of various diagnostic methods for lymph node metastasis (LNM) of papillary thyroid carcinoma (PTC) through network meta-analysis. Methods: In this experiment, databases such as CNKI, Wanfang, PubMed, and Web of Science were retrieved according to the Cochrane database, Prisma, and NMAP command manual. A meta-analysis was performed using STATA 15.0, and the value of the surface under the cumulative ranking curve (SUCRA) was used to determine the most effective diagnostic method. Quality assessments were performed using the Cochrane Collaboration's risk of bias tool, and publication bias was assessed using Deeks' funnel plot. Results: A total of 38 articles with a total of 6285 patients were included. A total of 12 diagnostic methods were used to study patients with LNM of PTC. The results showed that 12 studies were direct comparisons and 8 studies were indirect comparisons. According to the comprehensive analysis of the area of SUCRA, US+CT(86.8) had the highest sensitivity, FNAC had the highest specificity (92.4) and true positive predictive value (89.4), and FNAC+FNA-Tg had higher negative predictive value (99.4) and accuracy (86.8). In the non-invasive method, US+CT had the highest sensitivity, and the sensitivity (SEN) was [OR=0.59, 95% confidence interval (CI): (0.30, 0.89]. Among the invasive methods, the combined application of FNAC+FNA-Tg had higher diagnostic performance. The sensitivity was [OR=0.62, 95% CI: (0.26, 0.98)], the specificity (SPE) was [OR=1.12, 95% CI: (0.59, 1.64)], the positive predictive value was [OR=0.98, 95% CI: (0.59, 1.37)], the negative predictive value was [OR=0.64, 95% CI (0.38, 0.90)], and the accuracy was [OR=0.71, 95% CI: (0.31, 1.12)]. Conclusion: In the non-invasive method, the combined application of US+CT had good diagnostic performance, and in the invasive method, the combined application of FNAC+FNA-Tg had high diagnostic performance, and the above two methods were recommended.

5.
Angew Chem Int Ed Engl ; 60(36): 19779-19789, 2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34233057

RESUMO

We developed a cyclic amplification method for an organic afterglow nanoreporter for the real-time visualization of self-generated reactive oxygen species (ROS). We promoted semiconducting polymer nanoparticles (PFODBT) as a candidate for emitting near-infrared afterglow luminescence. Introduction of a chemiluminescent substrate (CPPO) into PFODBT (PFODBT@CPPO) resulted in a significant enhancement of afterglow intensity through the dual cyclic amplification pathway involving singlet oxygen (1 O2 ). 1 O2 produced by PFODBT@CPPO induced cancer cell necrosis and promoted the release of damage-related molecular patterns, thereby evoking immunogenic cell death (ICD)-associated immune responses through ROS-based oxidative stress. The afterglow luminescent signals of the nanoreporter were well correlated with light-driven 1 O2 generation and anti-cancer efficiency. This imaging strategy provides a non-invasive tool for predicting the therapeutic outcome that occurs during ROS-mediated cancer therapy.


Assuntos
Luminescência , Nanopartículas/metabolismo , Neoplasias/metabolismo , Polímeros/metabolismo , Humanos , Nanopartículas/química , Neoplasias/patologia , Polímeros/química , Espécies Reativas de Oxigênio/metabolismo , Oxigênio Singlete/química , Oxigênio Singlete/metabolismo
6.
Angew Chem Int Ed Engl ; 60(17): 9562-9572, 2021 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-33590957

RESUMO

Chemodynamic therapy is an emerging tumor therapeutic strategy. However, the anticancer effects are greatly limited by the strong acidity requirements for effective Fenton-like reaction, and the inevitably "off-target" toxicity. Herein, we develop an acidity-unlocked nanoplatform (FePt@FeOx @TAM-PEG) that can accurately perform the high-efficient and tumor-specific catalysis for anticancer treatment, through dual pathway of cyclic amplification strategy. Notably, the pH-responsive peculiarity of tamoxifen (TAM) drug allows for the catalytic activity of FePt@FeOx to be "turn-on" in acidic tumor microenvironments, while keeping silence in neutral condition. Importantly, the released TAM within cancer cells is able to inhibit mitochondrial complex I, leading to the upregulated lactate content and thereby the accumulated intracellular H+ , which can overcome the intrinsically insufficient acidity of tumor. Through the positive feedback loop, large amount of active FePt@FeOx nanocatalyzers are released and able to access to the endogenous H2 O2 , exerting the improved Fenton-like reaction within the more acidic condition. Finally, such smart nanoplatform enables self-boosting generation of reactive oxygen species (ROS) and induces strong intracellular oxidative stress, leading to the substantial anticancer outcomes in vivo, which may provide a new insight for tumor-specific cascade catalytic therapy and reducing the "off-target" toxicity to surrounding normal tissues.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Nanopartículas de Magnetita/química , Fotoquimioterapia , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima , Animais , Neoplasias da Mama/diagnóstico por imagem , Linhagem Celular Tumoral , Concentração de Íons de Hidrogênio , Camundongos , Estrutura Molecular , Estresse Oxidativo , Tamanho da Partícula , Espécies Reativas de Oxigênio/química
7.
Biosci Rep ; 40(8)2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32729621

RESUMO

Early childhood caries (ECC) is one of the most prevalent chronic infectious diseases in children. The effective prevention and treatment are heavy burdens and study hotspots for pediatric dentists. Many studies had investigated the relationship between preterm, low birth weight (LBW) and ECC; however, the results were inconsistent. The present study was conducted with an evidence-based study to figure out the relationship between preterm, LBW and ECC for the first time. After searching the database, case-control and cross-sectional studies relevant to the relationship between preterm, LBW and ECC up to December 2019 were included. The data about odds ratio (OR) and 95% confidence interval (95% CI) were extracted and calculated with STATA 14.0 Software. A total of 22 studies were included in this meta-analysis, 9 studies of which did not only explore the relationship between ECC with preterm, but also study the relationship between ECC and LBW, 7 studies of which explored the relationship between preterm and ECC, and 6 studies of which studied the relationship between LBW and ECC. The meta-analysis results showed that the preterm increased the risk of ECC (OR = 1.59, 95% CI: 1.36-1.87) significantly. There was no difference between LBW and normal birth weight in the incidence of ECC (OR = 1.12, 95% CI: 0.94-1.33). The meta-analysis results of adjustment OR about LBW were similar to the crude OR (OR = 1.05, 95% CI: 0.71-1.57). This meta-analysis indicated that preterm increased the risk of ECC significantly; however, LBW was not a risk factor for ECC.


Assuntos
Cárie Dentária/epidemiologia , Recém-Nascido de Baixo Peso , Recém-Nascido Prematuro , Nascimento Prematuro/epidemiologia , Fatores Etários , Peso ao Nascer , Estudos de Casos e Controles , Criança , Pré-Escolar , Estudos Transversais , Cárie Dentária/diagnóstico , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Masculino , Nascimento Prematuro/diagnóstico , Medição de Risco , Fatores de Risco
8.
Nano Lett ; 20(1): 176-183, 2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31777250

RESUMO

In chemodynamic therapy (CDT), real-time monitoring of reactive oxygen species (ROS) production is critical to reducing the nonspecific damage during CDT and feasibly evaluating the therapeutic response. However, CDT agents that can emit ROS-related signals are rare. Herein, we synthesize a semiconducting polymer nanoplatform (SPN) that can not only produce highly toxic ROS to kill cancer cells but also emit ROS-correlated chemiluminescent signals. Notably, the efficacy of both chemiluminescence and CDT can be significantly enhanced by hemin doping (∼10-fold enhancement for luminescent intensity). Such ROS-dependent chemiluminescence of SPN allows ROS generation within a tumor to be optically monitored during the CDT process. Importantly, SPN establishes an excellent correlation of chemiluminescence intensities with cancer inhibition rates in vitro and in vivo. Thus, our nanoplatform represents the first intelligent strategy that enables chemiluminescence-imaging-monitored CDT, which holds potential in assessing therapeutic responsivity and predicting treatment outcomes in early stages.


Assuntos
Medições Luminescentes , Neoplasias Experimentais , Fotoquimioterapia , Polímeros/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Microambiente Tumoral/efeitos dos fármacos , Animais , Linhagem Celular Tumoral , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias Experimentais/diagnóstico por imagem , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/metabolismo
9.
Lasers Med Sci ; 35(2): 473-485, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31691054

RESUMO

The objective of this study was to evaluate the clinical attachment level (CAL) gain of Er:YAG, Er,Cr; YSGG, Nd:YAG; and diode laser (DL) as monotherapy or adjunctive to scaling and root planing (SRP) of chronic periodontitis by network meta-analysis (NMA). Randomized controlled clinical trials (RCTs) about lasers applied in SRP of chronic periodontitis were searched from PubMed, Cochrane Library, Web of Science, Ovid, Science Direct, Wan Fang, and China National Knowledge Infrastructure (CNKI) databases up to September 2018 and from references of selected full-texts and related reviews. Standard mean differences and 95% confidence intervals were counted for CAL gain. The random effects NMA were performed in STATA software. There were 25 RCTs about CAL gain at 3 and/or 6 months after lasers were applied in SRP. No inconsistency was detected. Er:YAG as monotherapy gained significantly more CAL at 3 months than did SRP; no significant differences were detected among other comparisons. In terms of CAL gain at 3 months, the ranking result from best to worst was as follows: Er:YAG as monotherapy, DL adjunctive to SRP, Er:YAG adjunctive to SRP, Er,Cr;YSGG as monotherapy, Nd:YAG adjunctive to SRP, and SRP. In terms of CAL gain at 6 months, the ranking result from best to worst was as follows: DL adjunctive to SRP, Nd:YAG adjunctive to SRP, SRP, Er:YAG adjunctive to SRP, and Er:YAG as monotherapy. Laser-assisted periodontal treatment could be superior to SRP alone and could serve as a good adjunctive treatment tool.


Assuntos
Periodontite Crônica/terapia , Raspagem Dentária , Lasers Semicondutores/uso terapêutico , Metanálise em Rede , Perda da Inserção Periodontal/terapia , Aplainamento Radicular , China , Terapia Combinada , Humanos , Viés de Publicação , Ensaios Clínicos Controlados Aleatórios como Assunto , Risco , Resultado do Tratamento
10.
Artigo em Inglês | MEDLINE | ID: mdl-22341353

RESUMO

Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias/sangue , Proteína Amiloide A Sérica/isolamento & purificação , Acetonitrilas/química , Sequência de Aminoácidos , Biomarcadores Tumorais/química , Western Blotting , Precipitação Química , Cromatografia Líquida de Alta Pressão , Humanos , Dados de Sequência Molecular , Estadiamento de Neoplasias , Neoplasias/patologia , Isoformas de Proteínas , Proteína Amiloide A Sérica/análise , Proteína Amiloide A Sérica/biossíntese , Proteína Amiloide A Sérica/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
11.
Mol Cancer ; 8: 79, 2009 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-19785722

RESUMO

BACKGROUND: Thyroid carcinoma is the most common endocrine malignancy and a common cancer among the malignancies of head and neck. Noninvasive and convenient biomarkers for diagnosis of papillary thyroid carcinoma (PTC) as early as possible remain an urgent need. The aim of this study was to discover and identify potential protein biomarkers for PTC specifically. METHODS: Two hundred and twenty four (224) serum samples with 108 PTC and 116 controls were randomly divided into a training set and a blind testing set. Serum proteomic profiles were analyzed using SELDI-TOF-MS. Candidate biomarkers were purified by HPLC, identified by LC-MS/MS and validated using ProteinChip immunoassays. RESULTS: A total of 3 peaks (m/z with 9190, 6631 and 8697 Da) were screened out by support vector machine (SVM) to construct the classification model with high discriminatory power in the training set. The sensitivity and specificity of the model were 95.15% and 93.97% respectively in the blind testing set. The candidate biomarker with m/z of 9190 Da was found to be up-regulated in PTC patients, and was identified as haptoglobin alpha-1 chain. Another two candidate biomarkers (6631, 8697 Da) were found down-regulated in PTC and identified as apolipoprotein C-I and apolipoprotein C-III, respectively. In addition, the level of haptoglobin alpha-1 chain (9190 Da) progressively increased with the clinical stage I, II, III and IV, and the expression of apolipoprotein C-I and apolipoprotein C-III (6631, 8697 Da) gradually decreased in higher stages. CONCLUSION: We have identified a set of biomarkers that could discriminate PTC from non-cancer controls. An efficient strategy, including SELDI-TOF-MS analysis, HPLC purification, MALDI-TOF-MS trace and LC-MS/MS identification, has been proved successful.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Papilar/diagnóstico , Proteômica/métodos , Neoplasias da Glândula Tireoide/diagnóstico , Apolipoproteína C-I/sangue , Apolipoproteína C-I/química , Apolipoproteína C-III/sangue , Apolipoproteína C-III/química , Biomarcadores Tumorais/química , Carcinoma Papilar/sangue , Cromatografia Líquida de Alta Pressão/métodos , Haptoglobinas/química , Haptoglobinas/metabolismo , Humanos , Imunoensaio/métodos , Peso Molecular , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Neoplasias da Glândula Tireoide/sangue
12.
Proteomics ; 9(19): 4539-53, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19670372

RESUMO

Mitochondria play essential roles in cell physiological processes including energy production, metabolism, ion homeostasis, cell growth, aging and apoptosis. Proteomic strategies have been applied to the study of mitochondria since 1998; these studies have yielded decisive information about the diverse physiological functions of the organelle. As an ideal model biological system, the nematode Caenorhabditis elegans has been widely used in the study of several diseases, such as metabolic diseases and cancer. However, the mitochondrial proteome of C. elegans remains elusive. In this study, we purified mitochondria from C. elegans and performed a comprehensive proteomic analysis using the shotgun proteomic approach. A total of 1117 proteins have been identified with at least two unique peptides. Their physicochemical and functional characteristics, subcellular locations, related biological processes, and associations with human diseases, especially Parkinson's disease, are discussed. An orthology comparison was also performed between C. elegans and four other model organisms for a general depiction of the conservation of mitochondrial proteins during evolution. This study will provide new clues for understanding the role of mitochondria in the physiological and pathological processes of C. elegans.


Assuntos
Proteínas de Caenorhabditis elegans/análise , Caenorhabditis elegans , Mitocôndrias/química , Proteínas Mitocondriais/análise , Proteoma/análise , Animais , Caenorhabditis elegans/citologia , Caenorhabditis elegans/metabolismo , Cromatografia Líquida , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/ultraestrutura , Espectrometria de Massas em Tandem
13.
Proteome Sci ; 7: 7, 2009 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-19291297

RESUMO

BACKGROUND: Acute lymphoblastic leukemia (ALL) is a common form of cancer in children. Currently, bone marrow biopsy is used for diagnosis. Noninvasive biomarkers for the early diagnosis of pediatric ALL are urgently needed. The aim of this study was to discover potential protein biomarkers for pediatric ALL. METHODS: Ninety-four pediatric ALL patients and 84 controls were randomly divided into a "training" set (45 ALL patients, 34 healthy controls) and a test set (49 ALL patients, 30 healthy controls and 30 pediatric acute myeloid leukemia (AML) patients). Serum proteomic profiles were measured using surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy (SELDI-TOF-MS). A classification model was established by Biomarker Pattern Software (BPS). Candidate protein biomarkers were purified by HPLC, identified by LC-MS/MS and validated using ProteinChip immunoassays. RESULTS: A total of 7 protein peaks (9290 m/z, 7769 m/z, 15110 m/z, 7564 m/z, 4469 m/z, 8937 m/z, 8137 m/z) were found with differential expression levels in the sera of pediatric ALL patients and controls using SELDI-TOF-MS and then analyzed by BPS to construct a classification model in the "training" set. The sensitivity and specificity of the model were found to be 91.8%, and 90.0%, respectively, in the test set. Two candidate protein peaks (7769 and 9290 m/z) were found to be down-regulated in ALL patients, where these were identified as platelet factor 4 (PF4) and pro-platelet basic protein precursor (PBP). Two other candidate protein peaks (8137 and 8937 m/z) were found up-regulated in the sera of ALL patients, and these were identified as fragments of the complement component 3a (C3a). CONCLUSION: Platelet factor (PF4), connective tissue activating peptide III (CTAP-III) and two fragments of C3a may be potential protein biomarkers of pediatric ALL and used to distinguish pediatric ALL patients from healthy controls and pediatric AML patients. Further studies with additional populations or using pre-diagnostic sera are needed to confirm the importance of these findings as diagnostic markers of pediatric ALL.

14.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 23(6): 515-9, 2007 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-17553346

RESUMO

AIM: To find if human soluble tumor necrosis factor receptor II (p75) fused IgG Fc protein (sTNFR II-IgG Fc) could be expressed in Pichia pastoris with an active dimmer form and characterize its N-linked oligosaccharides. METHODS: Two gene fragment, human sTNFR II and IgGFc, were got by RT-PCR from leucocytes stimulated with LPS. And the chimeric gene sTNFR II-IgG Fc achieved through gene splicing by over lap extension (SOE) method was cloned into pPIC9 and transformed into methanotropic yeast Pichia pastoris. The fusion protein purified by Protein A affinity column was analyzed with SDS-PAGE electrophoresis under reducing or non-reducing conditions and immunological methods. The anti-TNF-alpha biological activity assay of fusion protein was performed with L929 cells and detected with MTT colorimetry. The N-linked oligosaccharides hydrolyzed from fusion protein were labeled with 8-amino-1, 3, 6-naphthalene trisulfonic acid (ANTS) were analyzed with fluorophore-assisted carbohydrate eletrophoresis (FACE) as well. RESULTS: The recombinant P. pastoris strain that expressed human sTNFR II-IgG Fc fusion protein was constructed. The expression level of fusion protein in 2 L flask reached 2 mg/L. SDS-PAGE and Western blot showed the expressed fusion protein purified by protein was a dimer linked with inter-molecular disulfide linkage. The fusion protein neutralized cytotoxic activity of TNF-alpha to L929 cells, and the EC(50) of the fusion protein to inhibit 5 x 10(4) U/L of TNF-alpha was 170 microg/L. The FACE analysis showed there are 11 to 13 hexoses on each N-linked oligosaccharide. CONCLUSION: The human sTNFR II-IgG Fc fusion protein is expressed successfully in P. pastoris and it could be a reference for the future expression of other Fc fusion proteins or immunoglobulins in Pichia pastoris.


Assuntos
Expressão Gênica , Imunoglobulina G/química , Imunoglobulina G/metabolismo , Pichia/metabolismo , Receptores do Fator de Necrose Tumoral/química , Receptores do Fator de Necrose Tumoral/metabolismo , Western Blotting , Dimerização , Eletroforese em Gel de Poliacrilamida , Etanercepte , Humanos , Imunoglobulina G/genética , Naftalenos/química , Oligossacarídeos/química , Pichia/genética , Receptores do Fator de Necrose Tumoral/genética
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