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1.
Inflamm Res ; 73(3): 363-379, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38189810

RESUMO

OBJECTIVE: Ferroptosis is a reactive oxygen species (ROS)- and iron-dependent form of non-apoptotic cell death process. Previous studies have demonstrated that ferroptosis participates in the development of inflammatory arthritis. However, the role of ferroptosis in rheumatoid arthritis (RA) inflammatory hypoxic joints remains unclear. This study sought to explore the underlying mechanism of ferroptosis on lipopolysaccharide (LPS)-induced RA fibroblast-like synoviocytes (FLSs). METHODS: FLSs, isolated from patients with RA, were treated with LPS and ferroptosis inducer (erastin and RSL-3), and ferroptosis inhibitor (Fer-1 and DFO), respectively. The cell viability was measured by CCK-8. The cell death was detected by flow cytometer. The proteins level were tested by Western blot. The cytosolic ROS and lipid peroxidation were determined using DCFH-DA and C11-BODIPY581/591 fluorescence probes, respectively. The small interfering RNA (siRNA) was used to knock down related proteins. The levels of malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), iron, inflammatory cytokines (IL6 and IL8), and LDH were analyzed by commercial kits. RESULTS: Ferroptosis was activated by LPS in RA FLS with increased cellular damage, ROS and lipid peroxidation, intracellular Fe and IL8, which can be further amplified by ferroptosis inducer (erastin and RSL-3) and inhibited by ferroptosis inhibitor (Fer-1 and DFO). Mechanistically, LPS triggered ferroptosis via NCOA4-mediated ferritinophagy in RA FLSs, and knockdown of NCOA4 strikingly prevent the process of ferroptosis. Intriguingly, LPS-induced RA FLSs became insensitive to ferroptosis and NCOA4-mediated ferritinophagy under hypoxia compared with normoxia. Knockdown of HIF-1α reverted ferroptosis and ferritinophagy evoking by LPS-induced RA FLSs inflammation under hypoxia. In addition, low dose of auranofin (AUR) induced re-sensitization of ferroptosis and ferritinophagy through inhibiting the expression of HIF-1α under hypoxia. CONCLUSIONS: NCOA4-mediated ferritinophagy was a key driver of ferroptosis in inflammatory RA FLSs. The suppression of NCOA4-mediated ferritinophagy protected RA FLSs from ferroptosis in LPS-induced inflammation under hypoxia. Targeting HIF-1α/NCOA4 and ferroptosis could be an effective and valuable therapeutic strategy for synovium hyperplasia in the patients with RA.


Assuntos
Artrite Reumatoide , Ferroptose , Sinoviócitos , Humanos , Lipopolissacarídeos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Interleucina-8/metabolismo , Artrite Reumatoide/metabolismo , Inflamação/induzido quimicamente , Inflamação/metabolismo , Hipóxia/metabolismo , Fatores de Transcrição/metabolismo , RNA Interferente Pequeno/genética , Fibroblastos/metabolismo , Ferro/metabolismo , Coativadores de Receptor Nuclear/genética , Coativadores de Receptor Nuclear/metabolismo
2.
Bioinformatics ; 38(Suppl 1): i53-i59, 2022 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-35758798

RESUMO

MOTIVATION: The presence of tumor cell clusters in pleural effusion may be a signal of cancer metastasis. The instance segmentation of single cell from cell clusters plays a pivotal role in cluster cell analysis. However, current cell segmentation methods perform poorly for cluster cells due to the overlapping/touching characters of clusters, multiple instance properties of cells, and the poor generalization ability of the models. RESULTS: In this article, we propose a contour constraint instance segmentation framework (CC framework) for cluster cells based on a cluster cell combination enhancement module. The framework can accurately locate each instance from cluster cells and realize high-precision contour segmentation under a few samples. Specifically, we propose the contour attention constraint module to alleviate over- and under-segmentation among individual cell-instance boundaries. In addition, to evaluate the framework, we construct a pleural effusion cluster cell dataset including 197 high-quality samples. The quantitative results show that the numeric result of APmask is > 90%, a more than 10% increase compared with state-of-the-art semantic segmentation algorithms. From the qualitative results, we can observe that our method rarely has segmentation errors.


Assuntos
Algoritmos , Derrame Pleural , Análise por Conglomerados , Humanos
3.
Cytometry A ; 99(7): 722-731, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33342063

RESUMO

Recent advances in the recognition of biological samples using machine vision have made this technology increasingly important in research and detection. Image segmentation is an important step in this process. This study focuses on how to reduce the interference factors such as the overlap between different types (or within the same type) of urine cells according to microfluidics and improve the machine vision segmentation accuracy for cell images. In this study, we demonstrate that the platform can realize this hypothesis using urine cell image segmentation as an example application. We first discuss the reported urine cell droplet microfluidic chip system, which can realize the test conditions in which urine cells are encapsulated in the droplet and isolated from salt crystallization and/or bacteria and other urine-formed elements. Then, based on the analysis conditions set in the aforementioned experiment, the proportions of red blood cells, white blood cells, and squamous epithelial cells covered by various formed elements in the total urine cells in the same urine sample are measured. We simultaneously analyze the percentage of urine cells covered by salt crystallization and the incidence of overlapping between urine cells. Finally, the Otsu algorithm is used to segment the urine cell images encapsulated by the droplet and the urine cell images not encapsulated by the droplet, and the Dice, Jaccard, precision, and recall values are calculated. The results suggest that the method of encapsulating single cells based on droplets can improve the image segmentation effect without optimizing the algorithm.


Assuntos
Microfluídica
4.
FASEB J ; 34(5): 6215-6228, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32232900

RESUMO

Microscopic polyangiitis (MPA) is a systemic autoimmune disease that primarily affects the small and medium blood vessels. Endothelial injury is one of the pathological hallmarks of MPA. However, the pathogenesis for this has not yet been fully elucidated. Exosomal microRNAs (miRNAs) have recently emerged as a new molecular pattern involved in the endothelial injury in other diseases. Hence, we speculated that MPA plasma-derived exosomes (MPA-exo) could induce the endothelial injury, which was likely to be aroused by the dysregulated exosomal miRNAs in MPA. In the present study, plasma-derived exosomes were isolated and identified. MPA-exo could be internalized by human renal glomerular endothelial cells (HRGECs) in vitro and induced HRGECs injury. Subsequently, a series of differentially expressed miRNAs in MPA-exo were identified by high-throughput sequencing analysis. Further bioinformatics analysis for the target genes of these differentially expressed miRNAs showed a potential mechanism for their possible role in MPA endothelial injury. Notably, we revealed a considerable correlation between miR-185-3p, miR-125a-3p, and clinical parameters. In conclusion, the current study revealed that differentially expressed miRNAs in MPA-exo are associated with the endothelial injury. Our results suggested that these miRNAs and their target genes might be involved in the inflammation process of MPA.


Assuntos
Células Endoteliais/patologia , Exossomos/genética , MicroRNAs/sangue , Poliangiite Microscópica/complicações , Doenças Vasculares/diagnóstico , Idoso , Estudos de Casos e Controles , Células Endoteliais/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Transdução de Sinais , Doenças Vasculares/sangue , Doenças Vasculares/etiologia
5.
Inflamm Res ; 68(7): 597-611, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31119302

RESUMO

OBJECTIVE: The present study was undertaken to validate whether TNF-α and calreticulin (CRT) serve as dual signaling to activate nucleotide-binding oligomerization domain-, leucine-rich repeat- and pyrin domain-containing 3 (NLRP3) inflammasome in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) and HUVECs. The effect of human antigen R (HuR) in NLRP3 inflammasome activation was also explored in RA FLS. METHODS: Immunofluorescence was used to determine the expression of NLRP3 and adaptor protein apoptosis associated speck-like protein containing a CARD (ASC) in RA synovial tissue and HuR location in RA FLS. Western blot and quantitative real-time PCR were employed to measure the priming effect of NLRP3 inflammasome in cells and HuR expression in synovial tissue. The concentrations of IL-1ß and IL-18 were detected by enzyme linked immunosorbent assay. Immunohistochemistry was used to visualize the expression of HuR in synovial tissue. HuR knockdown in RA FLS was achieved by siRNA-mediated gene silencing. RESULTS: Higher expression of NLRP3 and ASC in RA synovial tissue than those in osteoarthritis was detected. The staining of NLRP3, ASC and cleaved IL-1ß were observed in FLS and vascular endothelial cells in RA synovium. Expression of NLRP3 and pro-IL-1ß in RA FLS and HUVECs treated with TNF-α was increased. The pro-IL-18 expression was also enhanced in HUVECs, but not in RA FLS. TNF-α/CRT dual stimulation of cells gave rise to caspase-1 p20 expression and the secretion of IL-1ß. The secreted IL-18 was also elevated in HUVECs but not in RA FLS. HuR expression was significantly elevated in RA synovial tissue. TNF-α initiated the nucleocytoplasmic shuttling of HuR in both FLS and HUVECs. The knockdown of HuR in FLS incubated with TNF-α led to reduced caspase-1 p20 protein expression and further resulted in decreased secretion of IL-1ß in the presence of CRT. CONCLUSIONS: TNF-α/CRT dual signaling induced NLRP3 inflammasome activation, which could be suppressed by HuR knockdown presumably due to the block of HuR translocating from nucleus to cytoplasma.


Assuntos
Artrite Reumatoide/imunologia , Calreticulina/imunologia , Proteína Semelhante a ELAV 1/imunologia , Inflamassomos/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Fator de Necrose Tumoral alfa/imunologia , Células Endoteliais da Veia Umbilical Humana/imunologia , Humanos , Transdução de Sinais , Membrana Sinovial/imunologia , Sinoviócitos/imunologia
6.
J Cell Physiol ; 234(1): 837-848, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-30078183

RESUMO

Diabetes mellitus (DM) comprises a group of metabolic diseases characterized by insulin deficiency or resistance and hyperglycemia. We previously reported the presence of abnormal differentiation of small intestinal epithelial cells (IECs) in diabetic mice, but the exact mechanism of this phenomenon has not been thoroughly elucidated to date. In this study, we found that H19 was markedly upregulated in IECs of DM mice. H19 knockdown significantly inhibited abnormal differentiation of IECs in DM mice. Bioinformatics analysis identified miR-141-3p as a candidate for H19. Based on luciferase reporter assays, we found that miR-141-3p directly targeted H19. Luciferase reporter assays also showed that miR-141-3p could directly target ß-catenin. Furthermore, H19 might act as an endogenous "sponge" by competing for miR-141-3p binding to regulate miRNA targets in vitro and in vivo. In summary, our findings provide the first evidence supporting the role of H19 in IECs of DM mice, and miR-141-3p targets not only protein-coding genes but also the lncRNA H19.


Assuntos
Diabetes Mellitus/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , beta Catenina/genética , Animais , Diferenciação Celular/genética , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patologia , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Técnicas de Silenciamento de Genes , Humanos , Hiperglicemia/genética , Hiperglicemia/patologia , Resistência à Insulina/genética , Intestino Delgado/metabolismo , Intestino Delgado/patologia , Camundongos , Camundongos Endogâmicos NOD , Ligação Proteica
7.
Clin Exp Rheumatol ; 36(5): 841-849, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29652658

RESUMO

OBJECTIVES: Fibroblast-like synoviocytes (FLS) play key roles in synovium hyperplasia and pannus formation in rheumatoid arthritis (RA). The present study was undertaken to explore the mechanisms that calreticulin (CRT) promoted anti-apoptosis of RA FLS. METHODS: The expression of CRT and anti-apoptotic proteins Bcl-XL and Mcl-1 in RA synovium were detected by immunohistochemistry. The expression of Bcl-XL and Mcl-1 in RA FLS by CRT were determined. The phosphorylation of Akt and STAT3 was detected by western blot. The effect of CRT on proliferation of RA FLS was examined by MTT assay. The ability of CRT to inhibit RA FLS apoptosis was assessed by flow cytometry. RESULTS: Increased expressions of CRT, Bcl-XL and Mcl-1 were detected in RA synovium compared with osteoarthritis (OA). Moreover, CRT expression correlated positively with Bcl-XL and Mcl-1 in RA, respectively. In vitro, CRT induced upregulation of Bcl-XL and Mcl-1 protein levels in RA FLS, in dose/time dependent manners. Upregulated expression of Bcl-XL and Mcl-1 induced by CRT were inhibited by PI3K/Akt or STAT3 pathways inhibitors in RA FLS, respectively. The increased phosphorylation levels of Akt and STAT3 were also detected with CRT incubation, in dose/time dependent manners. Additionally, CRT rescued apoptosis of RA FLS mediated by FasL. CONCLUSIONS: This study showed that upregulation of Bcl-XL and Mcl-1 expression in RA FLS by CRT were PI3K/Akt and STAT3 signal pathways dependent, and promoted the anti-apoptosis of RA FLS. Therefore, this may represent a therapeutic target for the treatment of RA.


Assuntos
Apoptose/efeitos dos fármacos , Artrite Reumatoide/tratamento farmacológico , Calreticulina/farmacologia , Fibroblastos/efeitos dos fármacos , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Membrana Sinovial/efeitos dos fármacos , Sinoviócitos/efeitos dos fármacos , Proteína bcl-X/metabolismo , Artrite Reumatoide/enzimologia , Artrite Reumatoide/patologia , Calreticulina/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Fibroblastos/enzimologia , Fibroblastos/patologia , Humanos , Fosforilação , Membrana Sinovial/enzimologia , Membrana Sinovial/patologia , Sinoviócitos/enzimologia , Sinoviócitos/patologia , Regulação para Cima
8.
Biomed Eng Online ; 15: 14, 2016 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-26847685

RESUMO

BACKGROUND: Cervical cancer is the second leading cause of female-specific cancer-related deaths after breast cancer, especially in developing countries. However, the incidence of the disease may be significantly decreased if the patient is diagnosed in the pre-cancerous lesion stage or earlier. In recent years, computer-based algorithms are widely used in cervical cancer screening. Most of the proposed algorithms follow the procedure of segmentation, feature extraction, and then classification. Nevertheless, few of the existing segmentation methods are as flexible and robust as the human visual system, and the complexity of the algorithms makes it difficult for clinical application. METHODS: In this study, a computer-assisted analytical approach is proposed to identify the existence of suspicious cells in a whole slide cervical cell image (WSCCI). The main difference between our method and the conventional algorithm is that the image is divided into blocks with certain size instead of segmented cells, which can greatly reduce the computational complexity. Via data analysis, some texture and color histogram features show significant differences between blocks with and without suspicious cells. Therefore these features can be used as the input of the support vector machine classifier. 1100 non-background blocks (110 suspicious blocks) are trained to build a model, while 1040 blocks (491 non-background blocks) from 12 other WSCCIs are tested to verify the feasibility of the algorithm. RESULTS: The experimental results show that the accuracy of our method is about 98.98 %. More importantly, the sensitivity, which is more fatal in cancer screening, is 95.0 % according to the images tested in the study, while the specificity is 99.33 %. CONCLUSION: The analysis of the algorithm is based on block images, which is different from conventional methods. Although some analysis work should be done in advance, the later processing speed will be greatly enhanced with the establishment of the model. Furthermore, since the algorithm is based on the actual WSCCI, the method will be of directive significance for clinical screening.


Assuntos
Colo do Útero/patologia , Diagnóstico por Computador/métodos , Processamento de Imagem Assistida por Computador/métodos , Programas de Rastreamento , Imagem Molecular , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologia , Automação , Feminino , Humanos , Máquina de Vetores de Suporte
9.
Age Ageing ; 44(3): 458-64, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25673873

RESUMO

BACKGROUND: in vitro, it has been reported that amyloid ß (Aß) is bound to red blood cells (RBCs) and this process damages the red cell. Also, a possible relationship between RBCs and Alzheimer's disease (AD) is supported by the findings of RBC impairment in AD. Therefore, Aß fibrils bounding RBC are of great interest as potential biomarkers. METHODS: in this study, we focused on Aß amyloid fibrils and/or aggregation on the peripheral RBC from 50 subjects with AD and 50 healthy controls (HCs) through thioflavin T (ThT) staining followed by immunofluorescence assay to confirm the presence of Aß amyloid fibrils and/or aggregation on the RBC. Then we optimised fluorescence staining and imaging conditions and analysed the images obtained by image processing software. RESULTS: we have analysed RBC morphology in blood from 50 subjects with AD and 50 HCs found that 16.8% of the RBCs are elongated as compared with 6.7% in normal controls (P < 0.01), and there is a negative correlation between the two parameters (P < 0.05). Our study showed that 98% of AD peripheral RBCs were amyloid binding-positive (ranging from 2 to 30%), while only 38% that of RBCs (ranging from 2 to 3.4%) were in HCs. We also found four modified morphologies of RBCs triggered by Aß binding, which may serve as an ancillary investigation and indicate the progression of AD. CONCLUSION: we first directly prove the existence of Aß binding RBCs in peripheral blood. In addition, we observed new modified morphologies of RBC triggered by Aß binding, all of those can serve as a biomarker for the diagnosis and progression of AD.


Assuntos
Doença de Alzheimer/diagnóstico , Peptídeos beta-Amiloides/sangue , Eritrócitos/metabolismo , Idoso , Doença de Alzheimer/sangue , Peptídeos beta-Amiloides/metabolismo , Biomarcadores/sangue , Estudos de Casos e Controles , Imunofluorescência , Humanos , Testes Neuropsicológicos
10.
Zhongguo Dang Dai Er Ke Za Zhi ; 17(9): 918-21, 2015 Sep.
Artigo em Zh | MEDLINE | ID: mdl-26412170

RESUMO

OBJECTIVE: To screen biomarkers which can be used as an auxiliary method in the diagnosis of Henoch-Schönlein purpura (HSP) and to evaluate their diagnostic values by receiver operating characteristic (ROC) curve analysis. METHODS: A total of 127 children diagnosed with HSP between April 2012 and March 2014 were included in the HSP group and an equal number of healthy children were included in the control group. Twelve parameters, i.e., serum amyloid protein A (SAA), interleukin-6 (IL-6), immunoglobulins (IgA, IgG, IgM, and IgE), C-reactive protein (CRP), white blood cell (WBC) count, complements C3 and C4, anti-streptolysin O, and ferritin, were analyzed. The values of the screened biomarkers for diagnosis of HSP were assessed by ROC curve analysis. RESULTS: The HSP group had significantly higher levels of SAA, IL-6, CRP, WBC, IgA, and IgM than the control group (P<0.05). The areas under the ROC curve of SAA, IL-6, WBC, IgA, and IgM for the diagnosis of HSP were higher than 0.7 (P<0.05). The optimal cut-off values of SAA, IgA, IgM, WBC, and IL-6 for the diagnosis of HSP were 3.035 µg/mL, 1579.5 mg/L, 922.5 mg/L, 8.850 × 109/L, and 7.035 pg/mL, respectively; the corresponding sensitivities of the optimal cut-off values for the diagnosis of HSP were 95.1%, 75.6%, 72.3%, 78.0%, and 63.4%, respectively, and the corresponding specificities were 90.2%, 85.4%, 82.4%, 70.7%, and 80.5%, respectively. CONCLUSIONS: SAA, IgA, IgM, WBC, and IL-6 are valuable biomarkers for clinical diagnosis of HSP and among them SAA seems to be the best one.


Assuntos
Vasculite por IgA/diagnóstico , Adolescente , Biomarcadores/sangue , Proteína C-Reativa/análise , Criança , Pré-Escolar , Feminino , Humanos , Vasculite por IgA/sangue , Masculino , Curva ROC , Proteína Amiloide A Sérica/análise
11.
Micromachines (Basel) ; 15(8)2024 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-39203631

RESUMO

The clinical diagnosis of a malignant pleural effusion (MPE) is still based on the detection of tumor cells in the pleural effusion. The question of how to improve the efficiency and accuracy of detecting an MPE still remains. This study explores the use of microfluidic technology to concentrate cells in an MPE and achieved the detection of the cell marker TPN in the microarray capture area. TPN is a mitochondria-specific bio-probe that can identify tumor cells on the basis of differences in the mitochondrial potential. First, we designed a microfluidic chip to analyze its performance. The results show that when the total flow rate of the injected chip was 12 mL/h and the volume ratio of cell separation liquid to cell suspension was 1:1, the target cells (A549, MCF-7, and Hela) were enriched and the purity was improved to 98.7-99.3%. Finally, an MPE from cancer patients was used to detect the chip's ability to isolate and enrich tumor cells. Furthermore, the fluorescent identification of the TPN within the tumor cells was simultaneously achieved on the microfluidic chip. In conclusion, the potential to improve the efficiency of the clinical diagnosis of MPEs is provided by the chip structure and analysis conditions explored in this study.

12.
Micromachines (Basel) ; 14(4)2023 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-37421004

RESUMO

Malignant pleural effusion is a common clinical problem, which often occurs in cases of malignant tumors, especially in lung cancer. In this paper, a pleural effusion detection system based on a microfluidic chip, combined with specific tumor biomarker, hexaminolevulinate (HAL), used to concentrate and identify tumor cells in pleural effusion was reported. The lung adenocarcinoma cell line A549 and mesothelial cell line Met-5A were cultured as the tumor cells and non-tumor cells, respectively. The optimum enrichment effect was achieved in the microfluidic chip when the flow rates of cell suspension and phosphate-buffered saline achieved 2 mL/h and 4 mL/h, respectively. At the optimal flow rate, the proportion of A549 increased from 28.04% to 70.01% due to the concentration effect of the chip, indicating that tumor cells could be enriched by a factor of 2.5 times. In addition, HAL staining results revealed that HAL can be used to identify tumor cells and non-tumor cells in chip and clinical samples. Additionally, the tumor cells obtained from the patients diagnosed with lung cancer were confirmed to be captured in the microfluidic chip, proving the validity of the microfluidic detection system. This study preliminarily demonstrates the microfluidic system is a promising method with which to assist clinical detection in pleural effusion.

13.
Sci Rep ; 13(1): 2160, 2023 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-36750590

RESUMO

The accurate detection of leukocytes is the basis for the diagnosis of blood system diseases. However, diagnosing leukocyte disorders by doctors is time-consuming and requires extensive experience. Automated detection methods with high accuracy can improve detection efficiency and provide recommendations to inexperienced doctors. Current methods and instruments either fail to automate the identification process fully or have low performance and need suitable leukocyte data sets for further study. To improve the current status, we need to develop more intelligent strategies. This paper investigates fulfilling high-performance automatic detection for leukocytes using a deep learning-based method. We established a new dataset more suitable for leukocyte detection, containing 6273 images (8595 leukocytes) and considering nine common clinical interference factors. Based on the dataset, the performance evaluation of six mainstream detection models is carried out, and a more robust ensemble model is proposed. The mean of average precision (mAP) @IoU = 0.50:0.95 and mean of average recall (mAR)@IoU = 0.50:0.95 of the ensemble model on the test set are 0.853 and 0.922, respectively. The detection performance of poor-quality images is robust. For the first time, it is found that the ensemble model yields an accuracy of 98.84% for detecting incomplete leukocytes. In addition, we also compared the test results of different models and found multiple identical false detections of the models, then provided correct suggestions for the clinic.


Assuntos
Aprendizado Profundo , Redes Neurais de Computação , Leucócitos
14.
Mol Immunol ; 142: 83-94, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34971867

RESUMO

Rheumatoid arthritis (RA) synovium was identified as "tumor-like" tissues because of the hypoxic microenvironment, significant cell proliferation, and invasion phenotypes. It was reported that hypoxia promoted tumor aggressiveness via up-regulated expression of fascin-1 in cancer. However, the role of fascin-1 in RA synovial hyperplasia and joint injury progression remains unknown. In the current study, we first identified that both fascin-1 and HIF-1α were highly expressed in the RA synovium, in which they were widely colocalized, compared to osteoarthritis(OA). As well, levels of fascin-1 in RA fibroblast-like synoviocytes(FLSs) were found significantly higher than those in OA FLSs. Further, it was demonstrated that the mRNA and protein levels of fascin-1 in RA FLSs were up-regulated in hypoxia (3 % O2) and experimental hypoxia induced by cobalt chloride. Mechanistically, the HIF-1α-mediated hypoxia environment activated the gene expression of the fascin-1 protein, which in turn promoted the migration and invasion of RA FLSs. Accordingly, the restoration of FLSs migration and invasion was observed following siRNA-mediated silencing of fascin-1 and HIF-1α expression. Notably, under the experimental hypoxia, we found that the expression levels of fascin-1, HIF-1α, and p-STAT3 were increased in a time-dependent manner, and fascin-1and HIF-1α expressions were dependent on p-STAT3. Our results indicated that hypoxia-induced fascin-1 up-regulation promoted RA FLSs migration and invasion through the STAT3/HIF-1α/fascin-1 axis, which might represent a novel therapeutic target for the treatment of RA.


Assuntos
Artrite Reumatoide/patologia , Proteínas de Transporte/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Proteínas dos Microfilamentos/metabolismo , Fator de Transcrição STAT3/metabolismo , Membrana Sinovial/patologia , Sinoviócitos/patologia , Proteínas de Transporte/genética , Hipóxia Celular/fisiologia , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Células Cultivadas , Humanos , Hiperplasia/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Articulações/lesões , Proteínas dos Microfilamentos/genética , Osteoartrite/patologia , Interferência de RNA , RNA Interferente Pequeno/genética , Fator de Transcrição STAT3/genética , Membrana Sinovial/citologia
15.
Front Mol Biosci ; 9: 841209, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35463946

RESUMO

Background: Type 2 diabetes mellitus (T2DM) is a multifaceted disorder affecting epidemic proportion at global scope. Defective insulin secretion by pancreatic ß-cells and the inability of insulin-sensitive tissues to respond effectively to insulin are the underlying biology of T2DM. However, circulating biomarkers indicative of early diabetic onset at the asymptomatic stage have not been well described. We hypothesized that global and targeted mass spectrometry (MS) based metabolomic discovery can identify novel serological metabolic biomarkers specifically associated with T2DM. We further hypothesized that these markers can have a unique pattern associated with latent or early asymptomatic stage, promising an effective liquid biopsy approach for population T2DM risk stratification and screening. Methods: Four independent cohorts were assembled for the study. The T2DM cohort included sera from 25 patients with T2DM and 25 healthy individuals for the biomarker discovery and sera from 15 patients with T2DM and 15 healthy controls for the testing. The Pre-T2DM cohort included sera from 76 with prediabetes and 62 healthy controls for the model training and sera from 35 patients with prediabetes and 27 healthy controls for the model testing. Both global and targeted (amino acid, acylcarnitine, and fatty acid) approaches were used to deep phenotype the serological metabolome by high performance liquid chromatography-high resolution mass spectrometry. Different machine learning approaches (Random Forest, XGBoost, and ElasticNet) were applied to model the unique T2DM/Pre-T2DM metabolic patterns and contrasted with their effectiness to differentiate T2DM/Pre-T2DM from controls. Results: The univariate analysis identified unique panel of metabolites (n = 22) significantly associated with T2DM. Global metabolomics and subsequent structure determination led to the identification of 8 T2DM biomarkers while targeted LCMS profiling discovered 14 T2DM biomarkers. Our panel can effectively differentiate T2DM (ROC AUC = 1.00) or Pre-T2DM (ROC AUC = 0.84) from the controls in the respective testing cohort. Conclusion: Our serological metabolite panel can be utilized to identifiy asymptomatic population at risk of T2DM, which may provide utility in identifying population at risk at an early stage of diabetic development to allow for clinical intervention. This early detection would guide ehanced levels of care and accelerate development of clinical strategies to prevent T2DM.

16.
Metabolism ; 116: 154694, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33358943

RESUMO

BACKGROUND: Diet-induced obesity (DIO) is associated with chronic, low-grade inflammation in the hypothalamus. The inflammatory pathway of the hypothalamus is activated during obesity, and inhibition of activation of the inflammatory pathway can partially reverse obesity. Therefore, exploring new targets for inhibiting hypothalamic inflammation will provide new ideas for the prevention and treatment of obesity. Liver kinase B1 (LKB1), a serine/threonine kinase, is a tumor suppressor and metabolic regulator. Recent studies have shown that LKB1 has a certain anti-inflammatory effect. However, a role of LKB1 in the regulation of hypothalamic inflammation remains unclear. Therefore, we examined whether LKB1 overexpression in the hypothalamus could weaken the hypothalamic inflammation and inhibit the development of obesity. METHODS: LKB1 overexpressing adeno-associated virus (AAV) particles were injected stereotactically into the third ventricle (3 V) of C57BL/6 mice fed with HFD. We assessed changes in body mass and adiposity, food intake, hypothalamic inflammatory markers, and energy and glucose metabolism. RESULTS: LKB1 up-regulation in hypothalamus attenuated diet-induced hypothalamic inflammation, reduced food intake and body weight gain. In addition, the overexpression of hypothalamic LKB1 increased the insulin sensitivity and improved whole-body lipid metabolism, which attenuated hepatic fat accumulation and serum lipid levels. CONCLUSION: Hypothalamic LKB1 up-regulation attenuates hypothalamic inflammation, and protects against hypothalamic inflammation induced damage to melanocortin system, resulting in lower food intake and lower fat mass accumulation, which consequently protects mice from the development of obesity. Our data suggest LKB1 as a novel negative regulator of hypothalamic inflammation, and also a potentially important target for treating other inflammatory diseases.


Assuntos
Doenças Hipotalâmicas/genética , Inflamação/genética , Obesidade/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Quinases Ativadas por AMP , Animais , Dieta Hiperlipídica , Doenças Hipotalâmicas/prevenção & controle , Hipotálamo/metabolismo , Hipotálamo/patologia , Inflamação/prevenção & controle , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Obesidade/etiologia , Obesidade/prevenção & controle , Especificidade de Órgãos/genética , Proteínas Serina-Treonina Quinases/metabolismo , Regulação para Cima/genética , Aumento de Peso/genética
17.
Comput Biol Chem ; 94: 107559, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34412001

RESUMO

OBJECTIVE: Pleural fluid biomarkers are beneficial for the complementary diagnosis of pleural effusion etiologies. This study focuses on the multidimensional evaluation of deep learning to investigate the pleural effusion biomarkers value and the diagnostic utility of combining these markers, in distinguishing pleural effusion etiologies. METHODS: Pleural effusion were divided into three groups according to the diagnosis and treatment guidelines: malignant pleural effusion (MPE), parapneumonic effusion (PPE), and congestive heart failure (CHF). First, the value of the biomarker was analyzed by a receiver operating characteristic (ROC) curve. Then by utilizing deep learning and entropy weight method (EWM), the clinical value of biomarkers was computed multidimensionally for complementary diagnosis of pleural effusion diseases. RESULTS: There were significant differences in the six biomarkers, TP, ADA, CEA, CYFRA211, NSE, MNC% (p < 0.05) and no significant differences in three physical characteristics including color, transparency, specific gravity and six other biomarkers such as WBC, PNC%, MTC%, pH level, GLU, LDH (p > 0.05) among the three pleural effusion groups. The comprehensive test of pleural fluid biomarkers based on deep learning is of high accuracy. The clinical value of cytomorphology biomarkers WBC, MNC %, PNC %, MTC % was higher among pleural fluid biomarkers. CONCLUSION: The clinical value of multi-dimensional analysis of biomarkers by deep learning and entropy weight method is different from the ROC curve analysis. It is suggested that during the clinical examination process, more attention should be paid to the cell morphology biomarkers, but the physical properties of the pleural fluid are less clinical significance.


Assuntos
Biomarcadores Tumorais/análise , Derrame Pleural Maligno/diagnóstico , Humanos
18.
Biochem Biophys Res Commun ; 400(4): 559-62, 2010 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-20807507

RESUMO

Recent studies clearly demonstrated that several types of pathogenic amyloid proteins acted as agents that could transmit amyloidosis by means of a prion-like mechanism. Systemic AA amyloidosis is one of the most severe complications of chronic inflammatory disorders, particularly rheumatoid arthritis. It is well known that, similar to an infectious prion protein, amyloid-enhancing factor (AEF) acts as a transmissible agent in AA amyloidosis. However, how AEF transmits AA amyloidosis in vivo remained to be fully elucidated. In the present study, we focused on finding cell-free forms of AEF and its carriers in circulation by using the murine transfer model of AA amyloidosis. We first determined that circulating cell-free AEF existed in blood and plasma in mice with systemic AA amyloidosis. Second, we established that plasma exosomes containing AA amyloid oligomers derived from serum amyloid A had AEF activity and could transmit systemic AA amyloidosis via a prion-like mechanism. These novel findings should provide insights into the transmission mechanism of systemic amyloidoses.


Assuntos
Amiloidose/metabolismo , Exossomos/metabolismo , Príons/metabolismo , Proteína Amiloide A Sérica/metabolismo , Amiloidose/sangue , Animais , Glicoproteínas/sangue , Glicoproteínas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C3H
19.
Orthop Surg ; 12(4): 1253-1260, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32558212

RESUMO

OBJECTIVE: The aim of the study is to evaluate the expression of lysosome-associated protein transmembrane-4 (LAPTM4B) in human osteosarcoma tissue samples collected in our hospital, and to explore the possible correlations between the clinical pathological features of osteosarcoma patients and LAPTM4B expression. METHODS: Immunohistochemical (IHC) assays were performed to detect the expression levels of LAPTM4B in 62 tissue samples of osteosarcoma tissues and corresponding non-tumor tissues. According to LAPTM4B staining intensity in tumor tissues, osteosarcoma patients were classified into LAPTM4B high expression and low expression groups. In addition, the potential correlations between LAPTM4B expression levels and clinical pathological features were evaluated. In addition, we detected the effects of LAPTM4B on the proliferation and invasion of esteosarcoma cells through colony formation assay and transwell assay, respectively. We further explored the potential effects of LAPTM4B on tumor growth and metastasis using in vivo animal model. RESULTS: We revealed that LAPTM4B was highly expressed in human osteosarcoma tissues. We determined the significance between LAPTM4B and clinical features, including the tumor size (P = 0.004*) and the clinical stage (P = 0.035*) of osteosarcoma patients. Our results further demonstrated that ablation of LAPTM4B obviously blocked the proliferation and invasion of osteosarcoma cells in vitro and restrained tumor growth and metastasis in mice. CONCLUSION: We investigated the potential involvement of LAPTM4B in osteosarcoma progression and confirmed LAPTM4B as a novel therapeutic target for osteosarcoma.


Assuntos
Neoplasias Ósseas/metabolismo , Proteínas de Membrana/metabolismo , Proteínas Oncogênicas/metabolismo , Osteossarcoma/metabolismo , Animais , Biomarcadores Tumorais/metabolismo , Neoplasias Ósseas/patologia , Neoplasias Ósseas/cirurgia , Linhagem Celular Tumoral , Progressão da Doença , Humanos , Camundongos , Osteossarcoma/patologia , Osteossarcoma/cirurgia , Carga Tumoral
20.
Life Sci ; 258: 118204, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32763296

RESUMO

AIMS: Liver kinase B1 (LKB1) is a serine/threonine kinase. Although many biological functions of LKB1 have been identified, the role of hypothalamic LKB1 in the regulation of central energy metabolism and susceptibility to obesity is unknown. Therefore, we constructed POMC neuron-specific LKB1 knockout mice (PomcLkb1 KO) and studied it at the physiological, morphological, and molecular biology levels. MAIN METHODS: Eight-week-old male PomcLkb1 KO mice and their littermates were fed a standard chow fat diet (CFD) or a high-fat diet (HFD) for 3 months. Body weight and food intake were monitored. Dual-energy X-ray absorptiometry was used to measure the fat mass and lean mass. Glucose and insulin tolerance tests and serum biochemical markers were evaluated in the experimental mice. In addition, the levels of peripheral lipogenesis genes and central energy metabolism were measured. KEY FINDINGS: PomcLkb1 KO mice did not exhibit impairments under normal physiological conditions. After HFD intervention, the metabolic phenotype of the PomcLkb1 KO mice changed, manifesting as increased food intake and an enhanced obesity phenotype. More seriously, PomcLkb1 KO mice showed increased leptin resistance, worsened hypothalamic inflammation and reduced POMC neuronal expression. SIGNIFICANCE: We provide evidence that LKB1 in POMC neurons plays a significant role in regulating energy homeostasis. LKB1 in POMC neurons emerges as a target for therapeutic intervention against HFD-induced obesity and metabolic diseases.


Assuntos
Deleção de Genes , Neurônios/enzimologia , Obesidade/enzimologia , Pró-Opiomelanocortina/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Quinases Ativadas por AMP , Tecido Adiposo/patologia , Animais , Dieta Hiperlipídica , Epididimo/patologia , Comportamento Alimentar , Regulação da Expressão Gênica , Glucose/metabolismo , Hipotálamo/patologia , Inflamação/patologia , Leptina/metabolismo , Fígado/enzimologia , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Obesidade/sangue , Obesidade/patologia , Pró-Opiomelanocortina/genética , Aumento de Peso
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