RESUMO
Introduction Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) can be limited by the inadequacy of intact tissues, especially in patients with lymphoma, sarcoidosis, and lymph node tuberculosis. A novel technique called transbronchial node biopsy (TBNB) by forceps or cryoprobe has been proposed and studied to improve specimen quality and diagnostic yield. We performed a systematic review of studies describing the safety and sensitivity of EBUS-TBNB versus EBUS-TBNA in diagnosing intrathoracic lymphadenopathy/masses. Methods We systematically searched MEDLINE, Embase, Cochrane, and China National Knowledge Infrastructure to identify studies focusing on the application of EBUS-TBNB for diagnosis of intrathoracic lymphadenopathy. The quality of each study was evaluated using the QUADAS-2 tool. Using inverse-variance weighting, we performed a meta-analysis of diagnostic yield estimations. We also reviewed the complications related to the procedure. Results Thirteen studies were included in the final analysis. The meta-analysis yielded a pooled overall diagnostic yield of 77.80% (939/1207) for EBUS-TBNA and 86.01% (834/958) for EBUS-TBNB, with an inverse variance-weighted odds ratio of 3.13 (95% confidence interval [CI], 1.61-6.01; p=0.0008) and I2 of 82%. The pooled diagnostic yield of EBUS-TBNB versus EBUS-TBNA for the diagnosis of malignancy (including primary lung cancer and extrapulmonary malignancy), was 84.53% (590/698) for EBUS-TBNA and 90.84% (476/524) for EBUS-TBNB, with an I-V-weighted OR of 2.33 (95% CI, 1.15-4.74; p=0.02) and I2 of 64%. The pooled diagnostic yield of EBUS-TBNB versus EBUS-TBNA for the diagnosis of benignancy, was 71.19% (252/354) for EBUS-TBNA and 86.62% (233/269) for EBUS-TBNB, with an I-V-weighted OR of 4.39 (95% CI, 2.00-9.65; p=0.002) and I2 of 59%. The overall complications included bleeding (n=11, 0.90%), pneumomediastinum (n=6, 0.49%), pneumothorax (n=6, 0.49%), pneumonia (n=4, 0.33%), respiratory failure (n=1, 0.08%), and haemoptysis (n=1, 0.08%). The funnel plot analysis illustrated no major publication bias. Conclusions EBUS-TBNB improves the overall diagnostic yield of sampling intrathoracic lymphadenopathy and mass lesions relative to EBUS-TBNA. The complication rate of EBUS-TBNB is higher than that of EBUS-TBNA but reportedly lower than that of surgical biopsies.
RESUMO
BACKGROUND: Endoscopic techniques, including endobronchial ultrasound (EBUS) and endoscopic ultrasound (EUS), are used as the initial approach for the diagnosis and staging of lung cancer and the diagnosis of thoracic and abdominal lesions. Historically, the transvascular approach has been avoided because of concerns about bleeding. OBJECTIVES: This article is a systematic review of studies evaluating the feasibility and safety of transvascular needle aspiration (TVNA) under the guidance of EBUS or EUS in the diagnosis of thoracic and abdominal lesions. METHODS: We performed a systematic search of the MEDLINE, Embase, and Cochrane databases to identify studies evaluating the application of EBUS/EUS-guided TVNA (EBUS/EUS-TVNA) for lesions located at the contralateral side of the vessel for which the transvascular approach was the best puncture path. We performed a meta-analysis of diagnostic yield estimations. We also reviewed the complications related to the procedure. RESULTS: Eleven observational studies were included in the final analysis. Meta-analysis yielded a pooled overall diagnostic yield of 82.10% (95% confidence interval, 0.74-0.89) for TVNA, with an I2 value of 52%. No publication bias was detected by Egger's test (p = 0.8528). The overall complications included minor bleeding, minor hematoma, pseudo-aneurysm of the aorta, hemoptysis, acute hypoxic respiratory failure, and moderate bleeding. The major complication rate was 2.71%. CONCLUSIONS: EBUS/EUS-TVNA is feasible and probably safe when performed by experienced endoscopists in carefully selected patients. In view of the potential risks associated with the transvascular approach, especially the development of hematoma and pseudoaneurysm, the fanning technique was avoided, and the area of aspiration should be assessed by EUS for 3 min after each aspiration. Most importantly, EBUS/EUS-TVNA should only be performed if the results will impact the clinical management.
Assuntos
Endossonografia , Neoplasias Pulmonares , Humanos , Endossonografia/métodos , Estudos de Viabilidade , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico/efeitos adversos , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico/métodos , Neoplasias Pulmonares/patologia , Broncoscopia/métodos , Estadiamento de Neoplasias , Linfonodos/patologia , Mediastino/diagnóstico por imagemRESUMO
Megakaryocytes (MKs) are the unique non-pathological cells that undergo polyploidization in mammals. The polyploid formation is critical for understanding the MK biology, and transcriptional regulation is involved in the differentiation and maturation of MKs. However, little is known about the functions of transcriptional elongation factors in the MK polyploidization. In this study, we investigated the role of transcription elongation factor EloA in the polyploidy formation during the MK differentiation. We found that EloA was highly expressed in the erythroleukemia cell lines HEL and K562. Knockdown of EloA in HEL cell line was shown to impair the phorbol myristate acetate (PMA) induced polyploidization process, which was used extensively to model megakaryocytic differentiation. Selective over-expression of EloA mutants with Pol II elongation activity partially restored the polyploidization. RNA-sequencing revealed that knockdown of EloA decelerated the transcription of genes enriched in the ERK1/2 cascade pathway. The phosphorylation activity of ERK1/2 decreased upon the EloA inhibition, and the polyploidization process of HEL was hindered when ERK1/2 phosphorylation was inhibited by PD0325901 or SCH772984. This study evidenced a positive role of EloA in HEL polyploidization upon PMA stimulation through enhanced ERK1/2 activity.
Assuntos
Sistema de Sinalização das MAP Quinases , Megacariócitos , Diferenciação Celular , Humanos , Megacariócitos/metabolismo , Poliploidia , Acetato de Tetradecanoilforbol/metabolismo , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
To better treat patients with non-small cell lung cancer (NSCLC), the investigations on novel molecules affecting NSCLC progression are of vital importance. Long noncoding RNAs (lncRNAs) are identified as pivotal regulators that can affect the cellular activities of carcinomas. Long intergenic non-protein coding RNA 667 (LINC00667) is a newly found lncRNA, and its expression pattern and potent mechanisms are still obscure in NSCLC. Our study was the first to illustrate that LINC00667 was upregulated in NSCLC and LINC00667 silence refrained the proliferation, migration, and angiogenesis of NSCLC cells in vitro. In addition, vascular endothelial growth factor A (VEGFA) was modulated by LINC00667 at posttranscriptional level. Furthermore, mechanism experiments depicted that LINC00667 recruited eukaryotic translation initiation factor 4A3 (EIF4A3) to stabilize VEGFA messenger RNA. Eventually, rescue assays implied that LINC00667 modulated NSCLC progression via EIF4A3-stabilized VEGFA. Jointly, these findings hinted that LINC00667 was a tumor promoter in NSCLC, providing guidance for the exploration on NSCLC treatment.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , RNA Helicases DEAD-box/metabolismo , Fator de Iniciação 4A em Eucariotos/metabolismo , Neoplasias Pulmonares/genética , RNA Longo não Codificante/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Carcinoma Pulmonar de Células não Pequenas/irrigação sanguínea , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/irrigação sanguínea , Neoplasias Pulmonares/patologia , Neovascularização Patológica/genética , Estabilidade de RNA , RNA Mensageiro/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
BACKGROUND: Helicobacter pylori (H. pylori) infection remains a global public health issue, especially in Asia. Due to the emergence of antibiotic-resistant strains and the complexity of H. pylori infection, conventional vaccination is the best way to control the disease. Our previous study found that the N-acetyl-neuroaminyllactose-binding hemagglutinin protein (HpaA) is an effective protective antigen for vaccination against H. pylori infection, and intranasal immunization with the immunodominant HpaA epitope peptide (HpaA 154-171, P22, MEGVLIPAGFIKVTILEP) in conjunction with a CpG adjuvant decreased bacterial colonization in H. pylori-infected mice. However, to confer more robust and effective protection against H. pylori infection, an optimized delivery system is needed to enhance the P22-specific memory T cell response. RESULTS: In this study, an intranasal nanoemulsion (NE) delivery system offering high vaccine efficacy without obvious cytotoxicity was designed and produced. We found that this highly stable system significantly prolonged the nasal residence time and enhanced the cellular uptake of the epitope peptide, which powerfully boosted the specific Th1 responses of the NE-P22 vaccine, thus reducing bacterial colonization without CpG. Furthermore, the protection efficacy was further enhanced by combining the NE-P22 vaccine with CpG. CONCLUSION: This epitope-loaded nanoemulsion delivery system was shown to extend antigen release and elicit potent Th1 response, it is an applicable delivery system for intranasal vaccine against H. pylori.
Assuntos
Portadores de Fármacos , Epitopos , Infecções por Helicobacter , Helicobacter pylori/imunologia , Fatores de Transcrição/imunologia , Administração Intranasal , Animais , Antígenos de Bactérias/imunologia , Sistemas de Liberação de Medicamentos , Emulsões , Epitopos/administração & dosagem , Epitopos/imunologia , Feminino , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/prevenção & controle , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas , VacinasRESUMO
We generated a new Bmp2 conditional-knockout allele without a neo cassette that removes the Bmp2 gene from osteoblasts (Bmp2-cKO(ob)) using the 3.6Col1a1-Cre transgenic model. Bones of Bmp2-cKO(ob) mice are thinner, with increased brittleness. Osteoblast activity is reduced as reflected in a reduced bone formation rate and failure to differentiate to a mature mineralizing stage. Bmp2 in osteoblasts also indirectly controls angiogenesis in the periosteum and bone marrow. VegfA production is reduced in Bmp2-cKO(ob) osteoblasts. Deletion of Bmp2 in osteoblasts also leads to defective mesenchymal stem cells (MSCs), which correlates with the reduced microvascular bed in the periosteum and trabecular bones. Expression of several MSC marker genes (α-SMA, CD146 and Angiopoietin-1) in vivo, in vitro CFU assays and deletion of Bmp2 in vitro in α-SMA(+) MSCs support our conclusions. Critical roles of Bmp2 in osteoblasts and MSCs are a vital link between bone formation, vascularization and mesenchymal stem cells.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Células-Tronco Mesenquimais/metabolismo , Neovascularização Patológica/metabolismo , Osteoblastos/metabolismo , Animais , Diferenciação Celular , Células-Tronco Mesenquimais/fisiologia , Camundongos , Periósteo , Transdução de SinaisRESUMO
BACKGROUND & AIMS: Immunodominance is an important feature of antiviral, antitumor, and antibacterial cellular immune responses, but it is not well demonstrated in the immune responses against Helicobacter pylori. Antigen-specific CD4(+) T cells protect mice against infection with H pylori. We investigated the immunodominant CD4(+) T-cell response to neuraminyllactose-binding hemagglutinin (HpaA), which is a conserved, H pylori-specific colonization factor that is being investigated as an antigen for vaccination strategies. METHODS: HpaA-specific CD4(+) T cells were expanded with autologous peripheral blood mononuclear cells that had been incubated with recombinant HpaA and characterized using overlapping synthetic peptides. We compared the percentage of CD4(+) T cells with specificity for HpaA(88-100), restricted to HLA-DRB1*1501, among 59 H pylori-infected subjects with different gastric diseases. RESULTS: We identified and characterized several immunodominant CD4(+) T-cell epitopes derived from HpaA. The immunodominant CD4(+) T-cell responses specific to HpaA(88-100) were observed in most H pylori-infected individuals who expressed HLA-DRB1*1501 and were significantly more abundant in patients with less severe diseases (P < .05). CONCLUSIONS: The HLA-DRB1*1501-restricted immunodominant CD4(+) T-cell response to HpaA(88-100) is associated with reduced risk of severe gastric diseases. Further study of these and other immunodominant CD4(+) T-cell responses to H pylori will provide insight into mechanisms of protective immunity and aid in vaccine design.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Gastropatias/epidemiologia , Gastropatias/microbiologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Epitopos de Linfócito T/imunologia , Cadeias HLA-DRB1/imunologia , Humanos , Lectinas/imunologia , Lipoproteínas/imunologia , Risco , Gastropatias/prevenção & controleRESUMO
BACKGROUND AND PURPOSE: Is minimal residual disease (MRD) monitoring by multiparameter flow cytometry (MFC) prognostic for acute myeloid leukemia (AML) patients before allogeneic hemopoietic stem cell transplantation (allo-HSCT)? And if so, what level of MRD eradication can be used to help guide the timing of HSCT? Can haplo-HSCT improve the prognosis of AML patients with MRD positive? To figure out these questions, we initiated this retrospective study. METHODS: 96 AML patients were included retrospectively and divided into 5 groups, according to pre-transplantation MRD levels (from 5 × 10-2 to <1 × 10-4), to analyze the overall survival (OS), disease-free survival (DFS) and cumulative incidence of relapse (CIR). Secondly, we compared the prognosis of MRD-negative (MRDneg) and MRD-positive (MRDpos) AML patients (cutoff value = 1 × 10-3) who underwent allo-HSCT, and further analyzed the prognosis of MRDpos patients after received different transplantation modalities. RESULTS: It is found that the 2-year OS and DFS of MRD negative group were better than the MRD positive group, and that the deeper the eradication of MRD before transplantation, the better the prognosis of patients. The CIR in patients received HLA-identical transplantation, was higher in the MRDpos than in the MRDneg. Haploid transplantation reduced the CIR disparity between MRDpos and MRDneg group. Subsequently, in AML patients who remain MRD positive before HSCT, we show that haplo-HSCT offered a better prognosis than HLA-identical transplantation (MSDT and MUDT). CONCLUSION: It is suggested that achieving MFC-MRD <10-3 (10-4 or even better) before allo-HSCT could reduce the relapse of AML and improve OS and DFS significantly, while haplo-HSCT may be preferred for patients not achieving MRD negativity.
Assuntos
Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda , Citometria de Fluxo , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Leucemia Mieloide Aguda/etiologia , Leucemia Mieloide Aguda/terapia , Neoplasia Residual/etiologia , Prognóstico , Recidiva , Estudos Retrospectivos , Transplante HomólogoRESUMO
Tooth development is regulated by epithelial-mesenchymal interactions and their reciprocal molecular signaling. Bone morphogenetic protein 2 (Bmp2) is essential for tooth formation. However, the role of Bmp2 during enamel formation remains unknown in vivo. In this study, the role of Bmp2 in the regulation of postnatal enamel formation was investigated via the conditional ablation of Bmp2 in enamel using the (Osx-Cre) mouse. Bmp2 gene ablation was confirmed by PCR analysis in Osx-Cre, Bmp2(flox/flox) mice. Bmp2-null mice displayed a severe and profound tooth phenotype with asymmetric and open forked incisors. Microradiographs revealed broken incisor tips and dental pulp chamber exposure. The enamel layer of incisors and molars was thin with hypomineralization. Scanning electron microscopy analysis showed that the enamel surface was rough with chipping and the enamel lacked a typical prismatic architecture. These results demonstrate that Bmp2 is essential for enamel formation.
Assuntos
Proteína Morfogenética Óssea 2/deficiência , Esmalte Dentário/anormalidades , Animais , Densidade Óssea , Proteína Morfogenética Óssea 2/metabolismo , Esmalte Dentário/diagnóstico por imagem , Esmalte Dentário/patologia , Esmalte Dentário/ultraestrutura , Camundongos , Camundongos Knockout , Dente/diagnóstico por imagem , Dente/patologia , Dente/ultraestrutura , Microtomografia por Raio-XRESUMO
Synthesizing photo-realistic images based on text descriptions is a challenging task in the field of computer vision. Although generative adversarial networks have made significant breakthroughs in this task, they still face huge challenges in generating high-quality visually realistic images consistent with the semantics of text. Generally, existing text-to-image methods accomplish this task with two steps, that is, first generating an initial image with a rough outline and color, and then gradually yielding the image within high-resolution from the initial image. However, one drawback of these methods is that, if the quality of the initial image generation is not high, it is hard to generate a satisfactory high-resolution image. In this paper, we propose SAM-GAN, Self-Attention supporting Multi-stage Generative Adversarial Networks, for text-to-image synthesis. With the self-attention mechanism, the model can establish the multi-level dependence of the image and fuse the sentence- and word-level visual-semantic vectors, to improve the quality of the generated image. Furthermore, a multi-stage perceptual loss is introduced to enhance the semantic similarity between the synthesized image and the real image, thus enhancing the visual-semantic consistency between text and images. For the diversity of the generated images, a mode seeking regularization term is integrated into the model. The results of extensive experiments and ablation studies, which were conducted in the Caltech-UCSD Birds and Microsoft Common Objects in Context datasets, show that our model is superior to competitive models in text-to-image synthesis.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Redes Neurais de Computação , Semântica , CorRESUMO
Thrombocytopenia 2 (THC2) is one of the most prevalent forms of inherited thrombocytopenia. It is caused by a heterogeneous group of ANKRD26 gene mutation and shows a heterogeneous clinical and laboratory characteristics. We present a big Chinese family with 10 THC2 patients carrying c.-128G > T heterozygous substitution in the 5-untranslated region of the ANKRD26 gene. Although the platelets are fewer than 50 × 109/L in 8 THC2 family members, only the proband and her son show a higher WHO bleeding score. The proband and her son are also beta-thalassemia carriers with heterozygous c.52A > T mutation of HBB, which might not be associated with the increased bleeding tendency since 3 other family members with low bleeding tendency also carried both ANKRD26 c.-128G > T and HBB c.52A > T mutations. However, the proband and her son also show hypofibrinogenaemia, which is likely the cause of their more severe clinical manifestation. HID1 c.442G > T mutation was detected not only in these two hypofibrinogenaemia family members but also in the other 8 family members with normal blood fibrinogen levels. Our study suggests that the co-occurrence of other inherited genetic conditions associated with blood coagulation might contribute to the heterogeneity of clinical and laboratory characteristics in THC2 patients. Considering the hematologic and myeloid malignancy predisposition of THC2 patients and a large population of immune thrombocytopenia in China, we urge more attention to be paid to the diagnosis of THC2 patients to avoid misdiagnosis and mistreatment.
RESUMO
Thrombocytopenia 2 (THC2) is a major type of inherited thrombocytopenia caused by the persistent ANKRD26 expression during the late stage of megakaryocytopoiesis. For the first time, we generated a human induced pluripotent stem cell (hiPSC) line SHAMUi001-A from the bone marrow hematopoietic progenitor cells of a THC2 patient, who has a heterozygous mutation (c.-128G>T) in the 5'-UTR of ANKRD26 gene. SHAMUi001-A cells retain the mutation, display pluripotent stem cell characteristics, and have a normal female karyotype. This disease-specific hiPSC line will be a useful model for THC2 research.
Assuntos
Células-Tronco Pluripotentes Induzidas , Regiões 5' não Traduzidas , Quebra Cromossômica , Feminino , Heterozigoto , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Mutação/genéticaRESUMO
Generation of a floxed Bmp2/4 osteoblast cell line is a valuable tool for studying the modulatory effects of Bmp2 and Bmp4 on osteoblast differentiation as well as relevant molecular events. In this study, primary floxed Bmp2/4 mouse osteoblasts were cultured and transfected with simian virus 40 large T-antigen. Transfection was verified by polymerase chain reaction (PCR) and immunohistochemistry. To examine the characteristics of the transfected cells, morphology, proliferation and mineralization were analyzed, expression of cell-specific genes including Runx2, ATF4, Dlx3, Osx, dentin matrix protein 1, bone sialoprotein, osteopontin, osteocalcin, osteonectin and collagen type I was detected. These results show that transfected floxed Bmp2/4 osteoblasts bypassed senescence with a higher proliferation rate, but retain the genotypic and phenotypic characteristics similar to the primary cells. Thus, we for the first time demonstrate the establishment of an immortalized mouse floxed Bmp2/4 osteoblast cell line.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Proteína Morfogenética Óssea 4/metabolismo , Linhagem Celular Transformada , Osteoblastos/metabolismo , Fosfatase Alcalina/biossíntese , Animais , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 4/genética , Calcificação Fisiológica , Linhagem Celular , Proliferação de Células , Regulação da Expressão Gênica , Camundongos , Camundongos Mutantes , Osteoblastos/citologia , TransfecçãoRESUMO
HpaA is considered to be an effective protective antigen for vaccination against Helicobacter pylori (H. pylori) infection. Oral immunization with HpaA significantly decreases bacterial colonization in H. pylori infected mice. In this study, we investigated whether subcutaneous or intranasal immunization with HpaA could protect against H. pylori infection. Mice immunized subcutaneously with HpaA in Complete Freund's adjuvant, but not mice intranasally immunized with HpaA in CpG adjuvant acquired protection against H. pylori infection. However, intranasal immunization with immunodominant epitope peptides in CpG adjuvant protected mice against H. pylori infection, and immunodominant epitope peptides stimulated stronger Th1 responses and mediated more robust protection against H. pylori infection than subdominant ones. Our results suggest that the length of a candidate antigen is critical for particular vaccination routes, and that immunodominant epitope peptides are promising candidates for protection against H. pylori infection through nasal vaccination.
Assuntos
Infecções por Helicobacter/imunologia , Infecções por Helicobacter/prevenção & controle , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Epitopos Imunodominantes/imunologia , Peptídeos/administração & dosagem , Peptídeos/imunologia , Adjuvantes Imunológicos , Administração Intranasal , Animais , Anticorpos Antibacterianos/imunologia , Feminino , Citometria de Fluxo , Imunização/métodos , Camundongos , Camundongos Endogâmicos BALB C , Vacinação/métodosRESUMO
In mice, antigen-specific CD4+ T cell response is indispensible for the protective immunity against Helicobacter pylori (H. pylori). It has been demonstrated that neuraminyllactose-binding hemagglutinin (HpaA) immunization protected mice from H. pylori infection in a CD4+ T cell dependent manner. However, much remains unclear concerning the human CD4+ T cell responses to HpaA. We conducted a systematic study here to explore the immunodominant, HpaA-specific CD4+ T cell responses in H. pylori infected individuals. We found that HpaA-specific CD4+ T cell responses varied remarkably in their magnitude and had broad epitope-specificity. Importantly, the main responses focused on two regions: HpaA76-105 and HpaA130-159. The HLA-DRB1*0901 restricted HpaA142-159 specific CD4+ T cell response was the most immunodominant response at a population level. The immunodominant epitope HpaA142-159 was naturally presented and highly conserved. We also demonstrated that it was not the broad peptide specificity, but the strength of HpaA specific CD4+ T cell responses associated with gastric diseases potentially caused by H. pylori infection. Such investigation will aid development of novel vaccines against H. pylori infection.
Assuntos
Adesinas Bacterianas/imunologia , Linfócitos T CD4-Positivos/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Cadeias HLA-DRB1/análise , Infecções por Helicobacter/complicações , Humanos , Gastropatias/imunologiaRESUMO
Our previous studies have demonstrated that Fam20C promotes differentiation and mineralization of odontoblasts, ameloblasts, osteoblasts and osteocytes during tooth and bone development. Ablation of the Fam20C gene inhibits bone and tooth growth by increasing fibroblast growth factor 23 in serum and causing hypophosphatemia in conditional knockout mice. However, control and regulation of the expression of Fam20C are still unknown. In this study, we generated a transgenic reporter model which expresses green fluorescence protein (GFP) driven by the Fam20C promoter. Recombineering was used to insert a 16 kb fragment of the mouse Fam20C gene (containing the 15 kb promoter and 1.1 kb of exon 1) into a pBluescript SK vector with the topaz variant of GFP and a bovine growth hormone polyadenylation sequence. GFP expression was subsequently evaluated by histomorphometry on cryosections from E14 to adult mice. Fluorescence was evident in the bone and teeth as early as E17.5. The GFP signal was maintained stably in odontoblasts and osteoblasts until 4 weeks after birth. The expression of GFP was significantly reduced in teeth, alveolar bone and muscle by 8 weeks of age. We also observed colocalization of the GFP signal with the Fam20C antibody in postnatal 1- and 7-day-old animals. Successful generation of Fam20C-GFP transgenic mice will provide a unique model for studying Fam20C gene expression and the biological function of this gene during odontogenesis and osteogenesis.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Proteínas da Matriz Extracelular/genética , Odontogênese/genética , Osteogênese/genética , Animais , Proteínas de Fluorescência Verde/genética , Células HEK293 , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos TransgênicosRESUMO
Helicobacter pylori (H. pylori) infects more than half of the world's population, causing chronic gastritis, peptic ulcers and gastric cancer. Urease B subunit (UreB), a conserved protein of H. pylori, is capable of inducing specific CD4(+) T-cell responses and provides protection against this infection. Previous studies have confirmed the effectiveness of rUreB subunit vaccines in generating CD4(+) T-cell-mediated protection, but less is known regarding the roles of different subtypes of T-cell immunity, such as Th1, Th2 and Th17, particularly the immunodominant epitopes inducing specific CD4(+) T-cell responses, in vaccine-mediated protection. In this study, we demonstrated that the vaccination of BALB/c mice with rUreB resulted in significant antigen-specific Th1 and Th17 immune responses. Importantly, two novel Th epitopes, UreB317-329 and UreB409-421, which are recognized by a major population of CD4(+) T cells, were identified in immunized mice. Our results demonstrated that two novel epitopes can simultaneously induce Th1 and Th17 immune responses; however, only the epitope vaccine-induced CD4(+) T-cells secreting IFN-γ mediated the protection against H. pylori; cells secreting IL-17A did not. Taken together, our results suggest that two novel immunodominant epitopes can induce Th1 and Th17 immune responses, but only the induced Th1 lymphocytes mediate protection against H. pylori.
Assuntos
Infecções por Helicobacter/prevenção & controle , Helicobacter pylori/imunologia , Células Th1/fisiologia , Células Th17/fisiologia , Vacinação , Adjuvantes Imunológicos/administração & dosagem , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/química , Proteínas de Bactérias/imunologia , Células Cultivadas , Mapeamento de Epitopos , Epitopos de Linfócito T/imunologia , Feminino , Adjuvante de Freund/administração & dosagem , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/microbiologia , Interferon gama/metabolismo , Interleucina-17/metabolismo , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Células Th1/microbiologia , Células Th17/microbiologia , Urease/administração & dosagem , Urease/química , Urease/imunologiaRESUMO
Aim. To investigate the diagnostic yield and etiologies of patients with obscure gastrointestinal bleeding (OGIB) using capsule endoscopy (CE) or double-balloon enteroscopy (DBE). Method. We studied the data of 532 consecutive patients with OGIB that were referred to Xinqiao Hospital in Chongqing from December 2005 to January 2012. A lesion that was believed to be the source of the bleeding (ulceration, mass lesion, vascular lesion, visible blood, inflammation, or others) was considered to be a positive finding. We analyzed the diagnostic yield of CE and SBE and the etiologies of OGIB. Result. CE and SBE have similar diagnostic yields, at 71.9% (196/231) and 71.8% (251/304), respectively. The most common etiology was erosions/ulceration (27.1%) followed by mass lesion (19.4%) and angiodysplastic/vascular lesions (13.9%). By stratified analysis, we found that erosions/ulceration (27.1%) was the most common etiology for the 21-40-year age group. Mass lesion was the most common etiology in the 41-60-year age group. However, in the >60 years age group, angiodysplastic/vascular lesions were significantly increased compared with the other groups, even though erosions/ulceration was most common. Conclusion. In this study, we found that CE and SBE have similar diagnostic yields and erosions/ulceration was the most common reason for OGIB, followed by mass lesion and angiodysplasias.
RESUMO
Formation of the periodontium begins following onset of tooth-root formation in a coordinated manner after birth. Dental follicle progenitor cells are thought to form the cementum, alveolar bone and Sharpey's fibers of the periodontal ligament (PDL). However, little is known about the regulatory morphogens that control differentiation and function of these progenitor cells, as well as the progenitor cells involved in crown and root formation. We investigated the role of bone morphogenetic protein-2 (Bmp2) in these processes by the conditional removal of the Bmp2 gene using the Sp7-Cre-EGFP mouse model. Sp7-Cre-EGFP first becomes active at E18 in the first molar, with robust Cre activity at postnatal day 0 (P0), followed by Cre activity in the second molar, which occurs after P0. There is robust Cre activity in the periodontium and third molars by 2 weeks of age. When the Bmp2 gene is removed from Sp7(+) (Osterix(+)) cells, major defects are noted in root, cellular cementum and periodontium formation. First, there are major cell autonomous defects in root-odontoblast terminal differentiation. Second, there are major alterations in formation of the PDLs and cellular cementum, correlated with decreased nuclear factor IC (Nfic), periostin and α-SMA(+) cells. Third, there is a failure to produce vascular endothelial growth factor A (VEGF-A) in the periodontium and the pulp leading to decreased formation of the microvascular and associated candidate stem cells in the Bmp2-cKO(Sp7-Cre-EGFP). Fourth, ameloblast function and enamel formation are indirectly altered in the Bmp2-cKO(Sp7-Cre-EGFP). These data demonstrate that the Bmp2 gene has complex roles in postnatal tooth development and periodontium formation.
Assuntos
Proteína Morfogenética Óssea 2/genética , Odontogênese/genética , Ligamento Periodontal/crescimento & desenvolvimento , Raiz Dentária/crescimento & desenvolvimento , Actinas/análise , Fator 2 Ativador da Transcrição/genética , Fatores Etários , Ameloblastos/patologia , Amelogênese/genética , Animais , Moléculas de Adesão Celular/análise , Diferenciação Celular/genética , Cementogênese/genética , Cemento Dentário/patologia , Polpa Dentária/irrigação sanguínea , Corantes Fluorescentes , Proteínas de Fluorescência Verde , Masculino , Camundongos , Camundongos Knockout , Microvasos/patologia , Dente Molar/crescimento & desenvolvimento , Dente Serotino/crescimento & desenvolvimento , Fatores de Transcrição NFI/análise , Odontoblastos/patologia , Fator de Transcrição Sp7 , Células-Tronco/fisiologia , Fatores de Transcrição/genética , Fator A de Crescimento do Endotélio Vascular/análise , Dedos de Zinco/genéticaRESUMO
An epitope-based vaccine is a promising option for treating Helicobacter pylori (H. pylori) infection. Epitope mapping is the first step in designing an epitope-based vaccine. A pivotal role of CD4(+) T cells in protection against H. pylori has been accepted, but few Th epitopes have been identified. In this study, two novel UreB CD4(+) T cell epitopes were identified using PBMCs obtained from two H. pylori infected subjects. We determined the restriction molecules by antibody blocking and used various Epstein-Barr virus-transformed B lymphocyte cell lines (BLCLs) with different HLA alleles as APCs to present peptides to CD4(+) T cells. These epitopes were DRB1*1404-restricted UreB(373-385) and DRB1*0803-restricted UreB(438-452). The T cells specific to these epitopes not only recognized autologous DCs loaded with recombinant UreB but also those pulsed with H. pylori whole cell lysates, suggesting that these epitope peptides are naturally processed. These epitopes have important value for designing an effective H. pylori vaccine.