RESUMO
Differences in clinical characteristics of early-onset and late-onset severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in neonates remain unclear. This study aimed to determine whether there are differences in the main clinical, radiological, and laboratory features of early-onset and late-onset SARS-CoV-2 infections in neonates. This single-center, prospective cohort study enrolled neonates with SARS-CoV-2 infection from December 7, 2022, to January 3, 2023, and evaluated their clinical characteristics during hospitalization. All neonates (N = 58) infected with SARS-CoV-2 within 28 days of birth who were admitted to the neonatal intensive care unit of Taizhou Hospital were included. These neonates were classified into the early-onset (diagnosed within 7 days of birth) and late-onset (diagnosed more than 7 days after birth) groups. The symptoms, treatment, and prognosis of SARS-CoV-2 infection were the main study outcomes. The incidence of hospitalization attributable to SARS-CoV-2 infection was 10.6% (58 of 546 neonates) in Linhai. Sixteen (28%) of the 58 SARS-CoV-2 infections were early-onset cases, and 42 (72%) were late-onset cases. The common symptoms among the late-onset group were fever (p < 0.001) and cough (p < 0.001). Neonates with late-onset SARS-CoV-2 infection (p < 0.001) were significantly more likely to develop pneumonia. Conclusion: The clinical symptoms and rates of pneumonia caused by SARS-CoV-2 infection in neonates differed between the early-onset and late-onset groups. Different clinical management is necessary for neonates with early-onset and late-onset SARS-CoV-2 infections. What is Known: ⢠Neonates are susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). ⢠Differences in clinical characteristics of early-onset and late-onset SARS-CoV-2 infections in neonates remain unclear. What is New: ⢠Fever and cough were the most common symptoms among neonates with late-onset infection. ⢠Neonates with late-onset SARS-CoV-2 infection were more likely to develop pneumonia.
Assuntos
COVID-19 , Pneumonia , Complicações Infecciosas na Gravidez , Recém-Nascido , Humanos , Gravidez , Feminino , COVID-19/diagnóstico , COVID-19/epidemiologia , SARS-CoV-2 , Estudos Prospectivos , Tosse , Febre/etiologia , Complicações Infecciosas na Gravidez/diagnósticoRESUMO
Changes in mitochondrial membrane permeability are closely associated with mitochondria-mediated apoptosis. Antimicrobial peptides (AMPs), which have been found to enter cells to exert physiological effects, cause damage to the mitochondria. This paper reviews the molecular mechanisms of AMP-mediated apoptosis by changing the permeability of the mitochondrial membrane through three pathways: the outer mitochondrial membrane (OMM), inner mitochondrial membrane (IMM), and mitochondrial permeability transition pore (MPTP). The roles of AMPs in inducing changes in membrane permeability and apoptosis are also discussed. Combined with recent research results, the possible application prospects of AMPs are proposed to provide a theoretical reference for the development of AMPs as therapeutic agents for human diseases.
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Peptídeos Antimicrobianos , Membranas Mitocondriais , Humanos , Membranas Mitocondriais/metabolismo , Mitocôndrias/fisiologia , Apoptose/fisiologia , Permeabilidade , Proteínas de Transporte da Membrana Mitocondrial/metabolismoRESUMO
OBJECTIVE: Bone marrow-derived hematopoietic stem and progenitor cells (HSC/HPC) are critical to homeostasis and tissue repair. The aims of this study were to delineate the myelotoxicity of cigarette smoking (CS) in a murine model, to explore human adipose-derived stem cells (hASC) as a novel approach to mitigate this toxicity, and to identify key mediating factors for ASC activities. METHODS: C57BL/6 mice were exposed to CS with or without i.v. injection of regular or siRNA-transfected hASC. For in vitro experiments, cigarette smoke extract was used to mimic the toxicity of CS exposure. Analysis of bone marrow HPC was performed both by flow cytometry and colony-forming unit assays. RESULTS: In this study, we demonstrate that as few as 3 days of CS exposure results in marked cycling arrest and diminished clonogenic capacity of HPC, followed by depletion of phenotypically defined HSC/HPC. Intravenous injection of hASC substantially ameliorated both acute and chronic CS-induced myelosuppression. This effect was specifically dependent on the anti-inflammatory factor TSG-6, which is induced from xenografted hASC, primarily located in the lung and capable of responding to host inflammatory signals. Gene expression analysis within bone marrow HSC/HPC revealed several specific signaling molecules altered by CS and normalized by hASC. CONCLUSION: Our results suggest that systemic administration of hASC or TSG-6 may be novel approaches to reverse CS-induced myelosuppression.
Assuntos
Tecido Adiposo/metabolismo , Moléculas de Adesão Celular/metabolismo , Mielopoese , Fumar/efeitos adversos , Transplante de Células-Tronco , Células-Tronco/metabolismo , Tecido Adiposo/patologia , Animais , Moléculas de Adesão Celular/farmacologia , Modelos Animais de Doenças , Feminino , Xenoenxertos , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Fumar/patologia , Células-Tronco/patologiaRESUMO
Dipeptidylpeptidase (DPP) 4 has the potential to truncate proteins with a penultimate alanine, proline, or other selective amino acids at the N-terminus. DPP4 truncation of certain chemokines, colony-stimulating factors, and interleukins have recently been linked to regulation of hematopoietic stem/progenitor cells, more mature blood cells, and other cell types. We believe that the potential role of DPP4 in modification of many regulatory proteins, and their subsequent effects on numerous stem/progenitor and other cell-type functions has not been adequately appreciated. This review addresses the potential implications of the modifying effects of DPP4 on a large number of cytokines and other growth-regulating factors with either proven or putative DPP4 truncation sites on hematopoietic cells, and subsequent effects of DPP4-truncated proteins on multiple aspects of steady-state and stressed hematopoiesis, including stem/progenitor cell, and more mature cell, function.
Assuntos
Dipeptidil Peptidase 4/metabolismo , Hematopoese/fisiologia , Células-Tronco Hematopoéticas/metabolismo , Proteínas/metabolismo , Animais , Diferenciação Celular , Linhagem da Célula , Movimento Celular/fisiologia , Células-Tronco Embrionárias/metabolismo , Células-Tronco Hematopoéticas/citologia , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Leucócitos/imunologia , Leucócitos/metabolismo , Células Progenitoras Linfoides/metabolismo , Modelos Biológicos , Células Progenitoras Mieloides/metabolismoRESUMO
SIRT1, an NAD-dependent deacetylase, plays a role in regulation of autophagy. SIRT1 increases mitochondrial function and reduces oxidative stress, and has been linked to age-related reactive oxygen species (ROS) generation, which is highly dependent on mitochondrial metabolism. H2O2 induces oxidative stress and autophagic cell death through interference with Beclin 1 and the mTOR signaling pathways. We evaluated connections between SIRT1 activity and induction of autophagy in murine (m) and human (h) embryonic stem cells (ESCs) upon ROS challenge. Exogenous H2 O2 (1 mM) induced apoptosis and autophagy in wild-type (WT) and Sirt1-/- mESCs. High concentrations of H2O2 (1 mM) induced more apoptosis in Sirt1-/-, than in WT mESCs. However, addition of 3-methyladenine, a widely used autophagy inhibitor, in combination with H2O2 induced more cell death in WT than in Sirt1-/- mESCs. Decreased induction of autophagy in Sirt1-/- mESCs was demonstrated by decreased conversion of LC3-I to LC3-II, lowered expression of Beclin-1, and decreased LC3 punctae and LysoTracker staining. H2O2 induced autophagy with loss of mitochondrial membrane potential and disruption of mitochondrial dynamics in Sirt1-/- mESCs. Increased phosphorylation of P70/85-S6 kinase and ribosomal S6 was noted in Sirt1-/- mESCs, suggesting that SIRT1 regulates the mTOR pathway. Consistent with effects in mESCs, inhibition of SIRT1 using Lentivirus-mediated SIRT1 shRNA in hESCs demonstrated that knockdown of SIRT1 decreased H2O2-induced autophagy. This suggests a role for SIRT1 in regulating autophagy and mitochondria function in ESCs upon oxidative stress, effects mediated at least in part by the class III PI3K/Beclin 1 and mTOR pathways.
Assuntos
Autofagia/fisiologia , Células-Tronco Embrionárias/metabolismo , Mitocôndrias/fisiologia , Sirtuína 1/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Apoptose/fisiologia , Proteínas Reguladoras de Apoptose/metabolismo , Autofagia/efeitos dos fármacos , Autofagia/genética , Proteína Beclina-1 , Western Blotting , Linhagem Celular , Células Cultivadas , Células-Tronco Embrionárias/citologia , Humanos , Peróxido de Hidrogênio/farmacologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Potencial da Membrana Mitocondrial/genética , Potencial da Membrana Mitocondrial/fisiologia , Camundongos Knockout , Microscopia Confocal , Mitocôndrias/metabolismo , Oxidantes/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Interferência de RNA , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
Understanding the factors that regulate hematopoiesis opens up the possibility of modifying these factors and their actions for clinical benefit. DEK, a non-histone nuclear phosphoprotein initially identified as a putative proto-oncogene, has recently been linked to regulate hematopoiesis. DEK has myelosuppressive activity in vitro on proliferation of human and mouse hematopoietic progenitor cells and enhancing activity on engraftment of long-term marrow repopulating mouse stem cells, has been linked in coordinate regulation with the transcription factor C/EBPα, for differentiation of myeloid cells, and apparently targets a long-term repopulating hematopoietic stem cell for leukemic transformation. This review covers the uniqueness of DEK, what is known about how it now functions as a nuclear protein and also as a secreted molecule that can act in paracrine fashion, and how it may be regulated in part by dipeptidylpeptidase 4, an enzyme known to truncate and modify a number of proteins involved in activities on hematopoietic cells. Examples are provided of possible future areas of investigation needed to better understand how DEK may be regulated and function as a regulator of hematopoiesis, information possibly translatable to other normal and diseased immature cell systems.
Assuntos
Proteínas Cromossômicas não Histona/fisiologia , Proteínas de Ligação a DNA/fisiologia , Hematopoese/fisiologia , Células-Tronco Hematopoéticas/fisiologia , Proteínas Oncogênicas/fisiologia , Animais , Humanos , Proteínas de Ligação a Poli-ADP-Ribose , Proto-Oncogene MasRESUMO
PURPOSE OF REVIEW: Dipeptidyl peptidase 4 (DPP4, CD26) is a protease that cleaves selected amino acids at the N-terminal penultimate position and has the potential to alter the protein function. The regulation and roles of DPP4 activity are not well understood; therefore, the purpose of this review is to discuss the recent literature regarding DPP4 regulation, as well as the variety of molecules it may affect, and their potential clinical applications. RECENT FINDINGS: Recent insight into the number of proteins that have DPP4 sites, and how DPP4 truncation may alter hematopoiesis based on the protein full length vs. truncated state, has shown that DPP4 truncation of colony-stimulating factors (CSFs) alters their function and that the activity of these CSFs can be enhanced when DPP4 activity is inhibited. DPP4 inhibition has recently been used in a clinical trial to attempt to enhance the engraftment of cord blood cells, and an endogenous DPP4 inhibitor tissue factor pathway inhibitor has been discovered, increasing our understanding of the potential importance of DPP4. SUMMARY: DPP4 plays a role in regulating the activity of CSFs and other cytokines involved in hematopoiesis. This information may be useful for enhancing hematopoietic cell transplantation, blood cell recovery after stress, and for understanding the physiology and pathophysiology of blood and other cell systems.
Assuntos
Dipeptidil Peptidase 4/fisiologia , Hematopoese/fisiologia , Transplante de Células-Tronco Hematopoéticas , Fatores Estimuladores de Colônias/metabolismo , Inibidores da Dipeptidil Peptidase IV , HumanosRESUMO
OBJECTIVE: To describe cytokine release syndrome (CRS) associated with immune checkpoint inhibitors (ICIs) reported in the FDA Adverse Event Reporting System (FAERS). METHODS: We obtained ICIs adverse event (AE) reports from January 2011 to September 2023 from the FAERS database. The preferred term (PT) 'cytokine release syndrome' from the Medical Dictionary for Regulatory Activities (MedDRA) 26.1 was used to identify cases with ICIs-related CRS. The reporting odds ratio (ROR) of the disproportionality method was performed to quantify the association between CRS and ICIs treatment strategy. RESULTS: Three hundred and ninety-five cases were gathered. 42.03% of the patients were aged 18 to 65. Male patients outnumbered female patients (53.67% vs. 34.94%). The prevalent potential cancer types were lung cancer (33.42%) and skin cancer (20.51%). Japanese were responsible for the majority of ICIs-related CRS cases (176 cases). The combination of nivolumab and ipilimumab resulted in the most CRS cases (138 cases), and the ICIs combination therapy had the highest ROR signal value (ROR = 11.95 [10.14-14.06]). ICIs-related CRS had a median time to onset of 14 days (interquartile range [IQR] 7-43.25). CONCLUSIONS: ICIs-related CRS is an increasingly important immune-related AE. Our study provided helpful information to help medical professionals learn more about ICIs-related CRS.
RESUMO
SIRT1 is a founding member of a sirtuin family of 7 proteins and histone deacetylases. It is involved in cellular resistance to stress, metabolism, differentiation, aging, and tumor suppression. SIRT1(-/-) mice demonstrate embryonic and postnatal development defects. We examined hematopoietic and endothelial cell differentiation of SIRT1(-/-) mouse embryonic stem cells (ESCs) in vitro, and hematopoietic progenitors in SIRT1(+/+)(+/-), and (-/-) mice. SIRT1(-/-) ESCs formed fewer mature blast cell colonies. Replated SIRT1(-/-) blast colony-forming cells demonstrated defective hematopoietic potential. Endothelial cell production was unaltered, but there were defects in formation of a primitive vascular network from SIRT1(-/-)-derived embryoid bodies. Development of primitive and definitive progenitors derived from SIRT1(-/-) ESCs were also delayed and/or defective. Differentiation delay/defects were associated with delayed capacity to switch off Oct4, Nanog and Fgf5 expression, decreased ß-H1 globin, ß-major globin, and Scl gene expression, and reduced activation of Erk1/2. Ectopic expression of SIRT1 rescued SIRT1(-/-) ESC differentiation deficiencies. SIRT1(-/-) yolk sacs manifested fewer primitive erythroid precursors. SIRT1(-/-) and SIRT1(+/-) adult marrow had decreased numbers and cycling of hematopoietic progenitors, effects more apparent at 5%, than at 20%, oxygen tension, and these progenitors survived less well in vitro under conditions of delayed growth factor addition. This suggests a role for SIRT1 in ESC differentiation and mouse hematopoiesis.
Assuntos
Envelhecimento , Células-Tronco Embrionárias/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Hematopoese/fisiologia , Células-Tronco Hematopoéticas/metabolismo , Sirtuína 1/metabolismo , Animais , Western Blotting , Separação Celular , Células-Tronco Embrionárias/citologia , Citometria de Fluxo , Expressão Gênica , Células-Tronco Hematopoéticas/citologia , Imuno-Histoquímica , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Sirtuína 1/deficiência , Sirtuína 1/genéticaRESUMO
Fusarium oxysporum causes wilt disease, which causes huge economic losses to a wide range of agricultural cash crops. Antifungal peptide P852 is an effective biocide. However, the mechanism of direct inhibition of pathogenic fungus needs to be explored. The proteomics and transcriptomics results showed that P852 mainly affected intracellular pathways such as glucose metabolism, amino acid metabolism, and oxidoreductase activity in F. oxysporum. P852 disrupts the intracellular oxidative equilibrium in F. oxysporum, and transmission electron microscopy observed mitochondrial swelling, disruption of membrane structure, and leakage of contents. Decreased mitochondrial membrane potential, mitochondrial cytochrome c leakage, and reduced ATP production were also detected. These results suggest that P852 is able to simultaneously inhibit intracellular metabolism and disrupt the mitochondrial function of F. oxysporum, exerting its inhibitory effects in multiple pathways together. The present study provides some insights into the multitargeted mechanism of fungus inhibition of antifungal lipopeptide substances produced by Bacillus spp.
Assuntos
Antifúngicos , Fusarium , Antifúngicos/farmacologia , Antifúngicos/metabolismo , Peptídeos/farmacologia , Peptídeos/metabolismo , Glucose/metabolismo , Aminoácidos/metabolismo , Doenças das Plantas/microbiologiaRESUMO
Green pesticides are highly desirable, as they are environmentally friendly and efficient. In this study, the antifungal peptide P852 was employed to suppress Fusarium wilt in the Faba bean. The disease index and a range of physiological and metabolomic analyses were performed to explore the interactions between P852 and the fungal disease. The incidence and disease index of Fusarium wilt were substantially decreased in diseased Faba beans that were treated with two different concentrations of P852 in both the climate chamber and field trial. For the first time, P852 exhibited potent antifungal effects on Fusarium in an open field condition. To explore the mechanisms that underlie P852's antifungal effects, P852 treatment was found to significantly enhance antioxidant enzyme capacities including guaiacol peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), and the activities of antifungal enzymes including chitinase and ß-1,3-glucanase, as well as plant dry and fresh weights, and chlorophyll content compared to the control group (p ≤ 0.05). Metabolomics analysis of the diseased Faba bean treated with P852 showed changes in the TCA cycle, biological pathways, and many primary and secondary metabolites. The Faba bean treated with a low concentration of P852 (1 µg/mL, IC50) led to upregulated arginine and isoquinoline alkaloid biosynthesis, whereas those treated with a high concentration of P852 (10 µg/mL, MFC) exhibited enhanced betaine and arginine accumulation. Taken together, these findings suggest that P852 induces plant tolerance under Fusarium attack by enhancing the activities of antioxidant and antifungal enzymes, and restoring plant growth and development.
RESUMO
OBJECTIVE: To test the hypothesis that ß -glucan enhances protective qi (PQi), an important Chinese medicine (CM) concept which stipulates that a protective force circulates throughout the body surface and works as the first line of defense against "external pernicious influences". METHODS: A total of 138 participants with PQi deficiency (PQD) were randomized to receive ß -glucan (200 mg daily) or placebo for 12 weeks. Participants' PQi status was assessed every 2 weeks via conventional diagnosis and a standardized protocol from which a PQD severity and risk score was derived. Indices of participants' immune and general health status were also monitored, including upper respiratory tract infection (URTI), saliva secretory IgA (sIgA), and self-reported measures of physical and mental health (PROMIS). RESULTS: PQi status was not significantly different between the ß -glucan and placebo treatment groups at baseline but improved significantly in the ß -glucan (vs. placebo) group in a time-dependent manner. The intergroup differences [95% confidence interval (CI)] in severity score (scale: 1-5), risk score (scale: 0-1), and proportion of PQD participants (%) at finish line was 0.49 (0.35-0.62), 0.48 (0.35-0.61), and 0.36 (0.25-0.47), respectively. Additionally, ß -glucan improved URTI symptom (scale: 1-9) and PROMIS physical (scale: 16.2-67.7) and mental (scale: 21.2-67.6) scores by a magnitude (95% CI) of 1.0 (0.21-1.86), 5.7 (2.33-9.07), and 3.0 (20.37-6.37), respectively, over placebo. CONCLUSIONS: ß -glucan ameliorates PQi in PQD individuals. By using stringent evidence-based methodologies, our study demonstrated that Western medicine-derived remedies, such as ß -glucan, can be employed to advance CM therapeutics. (ClinicalTrial.Gov registry: NCT03782974).
Assuntos
beta-Glucanas , Adulto , Método Duplo-Cego , Humanos , Qi , Fatores de Risco , Autorrelato , beta-Glucanas/uso terapêuticoRESUMO
Lactic acid bacteria (LAB) are a kind of Gram-positive bacteria which can colonize in the biological gastrointestinal tract and play a variety of probiotic roles. LAB have a wide range of applications in industry, animal husbandry, planting, food safety, and medical science fields. Previous studies on LAB have typically concentrated on their effects on improving the digestion and absorption of the gastrointestinal tract, regulating the balance of the microflora, and inhibiting the production and accumulation of toxic substances. The resistance of LAB to cancer is a topic of growing interest and relevance. This paper provided a summary of bio-active substances of LAB when they act against cancer, as well as the safety of LAB in clinical cancer treatment. Moreover, this paper further discussed several possible directions for future research and the potential application of LAB as anti-cancer therapy.
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We reported previously that HIV-1 the Tat-interacting protein of 110 kDa (TIP110; P110(NRB)/SART3/p110) is important in regulation of hematopoiesis, and in maintaining pluripotent factor (NANOG, OCT4, and SOX2) expression in and pluripotency of human embryonic stem cells (hESCs). Here we show that TIP110 expression in hESCs line H9 and embryonal carcinoma cell line NT-2 is regulated by C-MYC expression in ESCs through an E box present in the TIP110 promoter region. Through up- and down- modulation of expression, TIP110 induces OCT4 exon 1a inclusion and exon 1b skipping in our OCT4 minigene model. Thus, TIP110 expression in ESCs regulates alternative splicing of OCT4, an event required for pluripotency of ESCs.
Assuntos
Processamento Alternativo , Antígenos de Neoplasias/metabolismo , Células-Tronco Embrionárias/metabolismo , Fator 3 de Transcrição de Octâmero/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas de Ligação a RNA/metabolismo , Antígenos de Neoplasias/biossíntese , Linhagem Celular Tumoral , Células HeLa , Humanos , Células-Tronco Pluripotentes , Regiões Promotoras Genéticas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/biossínteseRESUMO
Enhancement of hematopoietic recovery after radiation, chemotherapy, or hematopoietic stem cell (HSC) transplantation is clinically relevant. Dipeptidylpeptidase (DPP4) cleaves a wide variety of substrates, including the chemokine stromal cell-derived factor-1 (SDF-1). In the course of experiments showing that inhibition of DPP4 enhances SDF-1-mediated progenitor cell survival, ex vivo cytokine expansion and replating frequency, we unexpectedly found that DPP4 has a more general role in regulating colony-stimulating factor (CSF) activity. DPP4 cleaved within the N-termini of the CSFs granulocyte-macrophage (GM)-CSF, G-CSF, interleukin-3 (IL-3) and erythropoietin and decreased their activity. Dpp4 knockout or DPP4 inhibition enhanced CSF activities both in vitro and in vivo. The reduced activity of DPP4-truncated versus full-length human GM-CSF was mechanistically linked to effects on receptor-binding affinity, induction of GM-CSF receptor oligomerization and signaling capacity. Hematopoiesis in mice after radiation or chemotherapy was enhanced in Dpp4(-/-) mice or mice receiving an orally active DPP4 inhibitor. DPP4 inhibition enhanced engraftment in mice without compromising HSC function, suggesting the potential clinical utility of this approach.
Assuntos
Quimiocina CXCL12/metabolismo , Dipeptidil Peptidase 4/metabolismo , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Hematopoese/fisiologia , Radioterapia/efeitos adversos , Transdução de Sinais/fisiologia , Animais , Linhagem Celular , Primers do DNA/genética , Dipeptidil Peptidase 4/genética , Hematopoese/efeitos dos fármacos , Hematopoese/efeitos da radiação , Humanos , Imunofenotipagem , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais/genéticaRESUMO
Toll-like receptor (TLR) activation is important in immune responses and in differentiation of hematopoietic stem cells. We detected mRNA expression of TLRs 1, 2, 3, 5, and 6, but not TLRs 4, 7, 8, and 9 in murine (m)ESC line E14, and noted high cell surface protein expression of TLR2, but not TLR4, for mESC lines R1, CGR8, and E14. ESC lines were cultured in the presence of leukemia inhibitory factor (LIF). Pam(3)Cys enhanced proliferation and survival of the 3 ESC lines. In contrast, lipopolysaccharide (LPS) decreased proliferation and survival. Pam(3)Cys and LPS effects on proliferation and survival were blocked by antibody to TLR2, suggesting that effects of both Pam(3)Cys and LPS on these mESC lines were likely mediated through TLR2. E14 ESC line expressed MyD88. Pam(3)Cys stimulation of E14 ESCs was associated with induced NF-kappaB translocation, enhanced phosphorylation of IKK-alpha/beta, and enhanced mRNA, but not protein, expression of tumor necrosis factor-alpha, interferon-gamma, and IL-6. TLR2 activation by Pam(3)Cys or inhibition by LPS was not associated with changes in morphology or expression of alkaline phosphatase, Oct4, SSEA1, KLF4, or Sox2, markers of undifferentiated mESCs. Our studies identify TLR2 as present and functional in E14, R1, and CGR8 mESC lines.
Assuntos
Proliferação de Células , Citocinas/genética , Células-Tronco Embrionárias/efeitos dos fármacos , Fator Inibidor de Leucemia/farmacologia , NF-kappa B/metabolismo , Receptor 2 Toll-Like/fisiologia , Animais , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Células Cultivadas , Citocinas/metabolismo , Células-Tronco Embrionárias/metabolismo , Células-Tronco Embrionárias/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Fator 4 Semelhante a Kruppel , Camundongos , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/metabolismoRESUMO
Nuclear tumor suppressor p53 transactivates proapoptotic genes or antioxidant genes depending on stress severity, while cytoplasmic p53 induces mitochondrial-dependent apoptosis without gene transactivation. Although SIRT1, a p53 deacetylase, inhibits p53-mediated transactivation, how SIRT1 regulates these p53 multifunctions is unclear. Here we show that SIRT1 blocks nuclear translocation of cytoplasmic p53 in response to endogenous reactive oxygen species (ROS) and triggers mitochondrial-dependent apoptosis in mouse embryonic stem (mES) cells. ROS generated by antioxidant-free culture caused p53 translocation into mitochondria in wild-type mES cells but induced p53 translocation into the nucleus in SIRT1(-/-) mES cells. Endogenous ROS triggered apoptosis of wild-type mES through mitochondrial translocation of p53 and BAX but inhibited Nanog expression of SIRT1(-/-) mES, indicating that SIRT1 makes mES cells sensitive to ROS and inhibits p53-mediated suppression of Nanog expression. Our results suggest that endogenous ROS control is important for mES cell maintenance in culture.
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Apoptose/fisiologia , Núcleo Celular/metabolismo , Células-Tronco Embrionárias/metabolismo , Proteínas de Homeodomínio/metabolismo , Sirtuínas/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Núcleo Celular/genética , Células Cultivadas , Citoplasma/genética , Citoplasma/metabolismo , Células-Tronco Embrionárias/citologia , Proteínas de Homeodomínio/genética , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/metabolismo , Proteína Homeobox Nanog , Espécies Reativas de Oxigênio/metabolismo , Sirtuína 1 , Sirtuínas/genética , Ativação Transcricional/fisiologia , Proteína Supressora de Tumor p53/genética , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
OBJECTIVE: To explore whether the modified E7-HSP70, which has been introduced mutations in two zinc-binding motifs of E7, will eliminate its transformation potential and enhance the immunogenicity of fusion protein and repress E7 containing tumors with a low level of MHC-I molecules to lung metastatic in murine model. METHODS: In this study, we examined the transforming properties of mutant E7 oncoprotein by the soft agar colony-formation assays, explored the immunogenicity of modified E7-HSP70 gene by various cellular and humor immune responses and evaluated the effect of treating lung metastatic tumor with a low expressing MHC-I molecules by tumor challenge assay and therapeutic experiment. RESULTS: The mutant E7 oncoprotein has completely lost its transforming properties as measured in the soft agar colony-formation assays. Modified E7-HSP70 gene inducted stronger E7-specific cellular immune response than that induced by unmodified E7-HSP70. More importantly, the new construct significantly reduced the number of B16-HPV16E7 lung metastases. CONCLUSION: The modified E7-HSP70 gene may be as a powerful and safe DNA vaccine in controlling the hematogenous spread of HPV16E7-associated tumors with low expression of MHC-I molecules. In addition, the B16-HPV16E7 lung metastasis model can be used to test the efficacy of various E7-specific vaccines and immunotherapeutic strategies in settings more relevant to clinical requirements.
Assuntos
Vacinas Anticâncer/farmacologia , Proteínas de Choque Térmico HSP70/imunologia , Neoplasias Pulmonares/terapia , Melanoma Experimental/terapia , Proteínas Oncogênicas Virais/imunologia , Vacinas de DNA/farmacologia , Animais , Especificidade de Anticorpos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/genética , Vacinas Anticâncer/imunologia , Transformação Celular Viral/genética , Transformação Celular Viral/imunologia , Feminino , Proteínas de Choque Térmico HSP70/genética , Antígenos de Histocompatibilidade Classe I/biossíntese , Antígenos de Histocompatibilidade Classe I/genética , Interferon gama/biossíntese , Interferon gama/genética , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/secundário , Melanoma Experimental/imunologia , Melanoma Experimental/secundário , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Regulação para Cima , Vacinas de DNA/genética , Vacinas de DNA/imunologiaRESUMO
OBJECTIVE: To compare clinical therapeutic effects of abdominal acupuncture and traditional acupuncture on cervical spondylosis (CS). METHODS: Sixty-two cases of neck or nerve-root type CS were randomly divided into an observation group (n=32) treated by abdominal acupuncture at Zhongwan (CV 12), Guanyuan (CV 4) and others, and a control group (n=30) treated by traditional acupuncture at Fengchi (GB 20) and cervical Jiaji (EX-B 2), etc.. Simplified McGill Pain Questionnaire (MPQ) and clinical therapeutic effects were served as the objective indexes. Their clinical therapeutic effects were compared after the first session of treatment, at the end of therapeutic course and 3 months after the end of treatment. RESULTS: The two groups had a same effective rate of 100.0%. All items of MPQ in these two groups after treatment and 3 months after the end of treatment significantly improved, and in the observation group the differences in the PRI feeling score before and after the first treatment, and the difference of the total PRI scores after the first treatment, at the end of therapeutic course and 3 months after the end of treatment significantly improved as compared with the control group (P < 0.05). CONCLUSION: Abdominal acupuncture can better reduce the pain of the patient caused by CS, with transient pain-alleviating effect, but whether or not the clinical therapeutic effect of abdominal acupuncture is better than the traditional acupuncture still can not be proved.
Assuntos
Terapia por Acupuntura/métodos , Vértebras Cervicais , Osteofitose Vertebral/terapia , Abdome , Adulto , Idoso , Feminino , Humanos , Masculino , Medicina Tradicional Chinesa , Pessoa de Meia-IdadeRESUMO
Downregulation of the expression of major histocompatibility complex class I antigens on the surface of high-risk HPVs-transformed cells may contribute to their high tumorigenic potential, which enables them to escape immune recognition by cytotoxic T lymphocytes. In this study, we show that the viral E7 oncoprotein mediates transcriptional downregulation of the major histocompatibility complex (MHC) class I genes by targeting the class I promoter in HPV16 containing CaSki tumor cells. Using the chromatin immunoprecipitation assay, we demonstrated that HPV16E7 and specific HADCs, including HADC1, HADC2, and HADC8, are physically associated with the class I promoter and the histone of the class I promoter was deacetylated. Knocking down of HPV16E7 expression with the E7-specific small interfering RNA induced the release of HPV17E7 as well as HDAC1 and HDAC2 from the class I promoter. Furthermore, HPV16E7 siRNA resulted in a dramatic increase in histone acetylation. Importantly, MHC class I antigen expression was up-regulated on the surface of cells transfected with the E7 siRNA, but not on that of untransfected cells. Taken together, our results demonstrate that the HPV16E7 protein is associated with the MHC class I promoter and mediates MHC class I downregulation by repressing chromatin activation.