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We report compound heterozygous variants in TOE1 in siblings of Chinese origin who presented with dyskinesia and intellectual disabilities. Our report provides further information regarding the etiology and pathogenesis of pontocerebellar hypoplasia type 7 syndrome (PCH7). Clinical manifestations were obtained, and genomic DNA was collected from family members. Whole-exome and Sanger sequencing were performed to identify associated genetic variants. Bioinformatics analysis was conducted to identify and characterize the pathogenicity of the heterozygous variants. Following long-term rehabilitation, both siblings showed minimal improvement, and their condition tended to progress. Whole-exome sequencing revealed two unreported heterozygous variants, NM_025077: c.C553T (p.R185W) and NM_025077: c.G562T (p.V188L), in the TOE1 gene mapped to 1p34.1. Sanger sequencing confirmed that the two variants in the proband and her brother were inherited from their parents. The NM_025077: c.C553T (p.R185W) variant was inherited from the father, and the NM_025077: c.G562T (p.V188L) variant was inherited from the mother. Although the two variants in the TOE1 gene have not been reported previously, they were associated with PCH7 based on integrated analysis. Thus, our report contributes to our knowledge regarding the etiology and phenotype of PCH 7.
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Doenças Cerebelares , Deficiência Intelectual , Humanos , Masculino , Feminino , Mutação , Deficiência Intelectual/genética , China , Linhagem , Proteínas Nucleares/genéticaRESUMO
Heparanase has been identified as a universal tumor-associated antigen, but heparanase epitope peptides are difficult to recognize. Therefore, it is necessary to explore novel strategies to ensure efficient delivery to antigen-presenting cells. Here, we established a novel immunotherapy model targeting antigens to dendritic cell (DC) receptors using a combination of heparanase CD4+ and CD8+ T-cell epitope peptides to achieve an efficient cytotoxic T-cell response, which was associated with strong activation of DCs. First, pegylated poly(lactic-coglycolic acid) (PLGA) nanoparticles (NPs) were used to encapsulate a combined heparanase CD4+ and CD8+ T-cell epitope alone or in combination with Toll-like receptor 3 and 7 ligands as a model antigen to enhance immunogenicity. The ligands were then targeted to DC cell-surface molecules using a DEC-205 antibody. The binding and internalization of these PLGA NPs and the activation of DCs, the T-cell response and the tumor-killing effect were assessed. The results showed that PLGA NPs encapsulating epitope peptides (mHpa399 + mHpa519) could be targeted to and internalized by DCs more efficiently, stimulating higher levels of IL-12 production, T-cell proliferation and IFN-γ production by T cells in vitro. Moreover, vaccination with DEC-205-targeted PLGA NPs encapsulating combined epitope peptides exhibited higher tumor-killing efficacy both in vitro and in vivo. In conclusion, delivery of PLGA NP vaccines targeting DEC-205 based on heparanase CD4+ and CD8+ T-cell epitopes are suitable immunogens for antitumor immunotherapy and have promising potential for clinical applications.
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Nanopartículas , Neoplasias , Humanos , Epitopos de Linfócito T/metabolismo , Ácido Poliglicólico/química , Ácido Poliglicólico/metabolismo , Receptor 3 Toll-Like , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/metabolismo , Ácido Láctico/química , Ácido Láctico/metabolismo , Ligantes , Células Dendríticas , Imunoterapia/métodos , Linfócitos T CD8-Positivos , Interleucina-12/metabolismo , Peptídeos/metabolismo , Linfócitos T CD4-Positivos , PolietilenoglicóisRESUMO
Serological tumor markers are useful for the detection of malignancies and evaluation of disease progression. These markers are not checked as part of a routine examination for patients with benign diseases and without any clinical suspicion of malignancy. However, some markers appear to be elevated in patients with benign diseases and without malignancies. We present a case of pyogenic liver abscesses with an elevated serum carcinoembryonic antigen (CEA) level associated with neither evidence of malignancy nor elevation of other tumor markers such as carbohydrate antigen (CA 19-9) and alpha-fetoprotein (AFP) levels. The serological level of CEA decreased and subsequently became within normal limits with treatment. This case also demonstrates that diabetic patients with a liver abscess may present with no infectious symptoms and that fine-needle aspiration is as effective as catheter drainage in the treatment of pyogenic liver abscess.
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BACKGROUND & AIMS: Breast regression protein 39 (BRP39) (Chi3L1) and its human homolog YKL-40, is an established biomarker of liver fibrosis in nonalcoholic steatohepatitis (NASH) patients, but its role in NASH pathogenesis remains unclear. We recently identified Chi3L1 as one of the top up-regulated genes in mice with inducible gain-of-function NOD-like receptor protein 3 (NLRP3) activation that mimics several liver features of NASH. This study aimed to investigate the effects of BRP39 deficiency on NLRP3-induced liver inflammation using tamoxifen-inducible Nlrp3 knockin mice sufficient (Nlrp3A350V CRT) and deficient for BRP39 (Nlrp3A350V/BRP-/- CRT). METHODS: Using Nlrp3A350V CRT mice and Nlrp3A350V BRP-/- CRT, we investigated the consequences of BRP39 deficiency influencing NLRP3-induced liver inflammation. RESULTS: Our results showed that BRP39 deficiency in NLRP3-induced inflammation improved body weight and liver weight. Moreover, liver inflammation, fibrosis, and hepatic stellate cell activation were reduced significantly, corresponding to significantly decreased Ly6C+ infiltrating macrophages, CD68+ osteopontin-positive hepatic lipid-associated macrophages, and activated Lymphocyte antigen 6 complex locus G6D positive (Ly6G+) and citrullinated histone H3 postivie (H3Cit+) neutrophil accumulation in the liver. Further investigation showed that circulatory neutrophils from NLRP3-induced BRP39-deficient mice have impaired chemotaxis and migration ability, and this was confirmed by RNA bulk sequencing showing reduced immune activation, migration, and signaling responses in neutrophils. CONCLUSIONS: These data showcase the importance of BRP39 in regulating the NLRP3 inflammasome during liver inflammation and fibrotic NASH by altering cellular activation, recruitment, and infiltration during disease progression, and revealing BRP39 to be a potential therapeutic target for future treatment of inflammatory NASH and its associated diseases.
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Hepatite , Hepatopatia Gordurosa não Alcoólica , Animais , Humanos , Camundongos , Fibrose , Inflamassomos/metabolismo , Inflamação/metabolismo , Infiltração de Neutrófilos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteínas NLR , Hepatopatia Gordurosa não Alcoólica/patologiaRESUMO
Despite the improved survival rate among systemic lupus erythematosus (SLE) patients, there are many factors associated with the mortality of SLE. In the current study, death-related factors of patients associated with course of disease were surveyed. Retrospective study was used. Mortalities among these three groups (group A, B and C, the course of disease was ≤ 5 years, 5-10 years and > 10 years, respectively) were calculated and compared. Various factors related to mortality were analyzed. Male SLE patients died relatively more than female patients. The total mortality was 8.5 %. The mortalities were significant difference in group A, B and C which were 9.4, 4.8 and 8.9 %, respectively. The mortalities of group A and group C were significantly higher than that of group B, but there was no significant difference between mortalities of group A and group C. The most common death-related factor was infection, followed by involved disorders in renal, brain, multisystem, heart, etc. The mortalities resulted from neuropsychiatric systemic lupus erythematosus (NPSLE), pulmonary infection, involved digestive system and hematological system were significantly different between three groups. There was no difference between mortalities of group A and group C associated with pulmonary infection and NPSLE. Patients in group C died more than in group A from involved renal, heart, multisystem, etc, while group A had more patients than group C who died of pulmonary infection, involved hematological system. In conclusion, Male SLE patients have worse outcome than female patients. Infection and active SLE are not only contributors to the death of early stage patients, but also to that of later stage patients.
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Lúpus Eritematoso Sistêmico/mortalidade , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Causas de Morte , Feminino , Humanos , Pacientes Internados , Vasculite Associada ao Lúpus do Sistema Nervoso Central/mortalidade , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores SexuaisRESUMO
In recent years, there has been a rapid expansion in our understanding of regulated cell death, leading to the discovery of novel mechanisms that govern diverse cell death pathways. One recently discovered type of cell death is pyroptosis, initially identified in the 1990s as a caspase-1-dependent lytic cell death. However, further investigations have redefined pyroptosis as a regulated cell death that relies on the activation of pore-forming proteins, particularly the gasdermin family. Among the key regulators of pyroptosis is the inflammasome sensor NOD-like receptor 3 (NLRP3), a critical innate immune sensor responsible for regulating the activation of caspase-1 and gasdermin D. A deeper understanding of pyroptosis and its interplay with other forms of regulated cell death is emerging, shedding light on a complex regulatory network controlling pore-forming proteins and cell fate. Cell death processes play a central role in diseases such as metabolic dysfunction-associated steatotic liver disease, metabolic dysfunction-associated steatohepatitis, autoinflammatory disorders, and cancer. Cell death often acts as a starting point in these diseases, making it an appealing target for drug development. Yet, the complete molecular mechanisms are not fully understood, and new discoveries reveal promising novel avenues for therapeutic interventions. In this review, we summarize recent evidence on pathways and proteins controlling pyroptosis and gasdermins. Furthermore, we will address the role of pyroptosis and the gasdermin family in metabolic dysfunction-associated steatotic liver disease and steatohepatitis. Additionally, we highlight new potential therapeutic targets for treating metabolic dysfunction-associated steatohepatitis and other inflammatory-associated diseases.
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Chitinase-3-like 1 protein (CHI3L1) is a secreted glycoprotein, strongly correlated with fibrosis severity in chronic liver diseases including non-alcoholic steatohepatitis (NASH). However, the mechanisms by which CHI3L1 contributes to fibrogenesis remain undefined. Here, we showed that infiltrating monocyte-derived liver macrophages represent the main source of CHI3L1 in murine NASH. We developed a floxed CHI3L1 knock-out (KO) mouse to further study the cell-specific role of CHI3L1 ablation. Wildtype (WT) and myeloid cell-specific CHI3L1 KO mice (CreLyz) were challenged with a highly inflammatory and fibrotic dietary model of NASH by administering choline-deficient high-fat diet for 10 weeks. Macrophage accumulation and inflammatory cell recruitment were significantly ameliorated in the CreLyz group compared to WT (F4/80 IHC p < 0.0001, CD11b IHC p < 0.0001). Additionally, hepatic stellate cell (HSC) activation and fibrosis were strongly decreased in this group (α-SMA IHC p < 0.0001, picrosirius red staining p < 0.0001). In vitro studies were performed stimulating bone marrow derived macrophages, THP-1 (human monocytes) and LX2 (human HSCs) cells with recombinant CHI3L1 to dissect its relationship with fibrosis development. Results showed an important role of CHI3L1 regulating fibrosis-promoting factors by macrophages (TGFB1 p < 0.05, CTGF p < 0.01) while directly activating HSCs (ACTA2 p < 0.01, COL1A1 p < 0.01), involving IL13Rα2 as the potential mediator. Our findings uncovered a novel role of CHI3L1 derived from liver macrophages in NASH progression and identifies this protein as a potential anti-fibrotic therapeutic target. KEY MESSAGES: We showed that CHI3L1 expression is increased in murine CDAA-HFAT diet NASH model, and that infiltrating macrophages are a key source of CHI3L1 production. Myeloid cell-specific CreLyz CHI3L1 knock-out in mice fed with CDAA-HFAT diet improved the NASH phenotype, with significantly reduced accumulation of pro-inflammatory macrophages and neutrophils compared with WT group. DEG and qPCR analysis of genes in CreLyz CHI3L1 knock-out mouse liver showed the mechanistic role of CHI3L1 in cellular chemotaxis. HSC is directly activated by CHI3L1 via receptor IL13Rα2, leading to upregulation of collagen deposition and pro-fibrotic gene, TIMP-1 and TIMP-2 release in whole liver. Direct stimulation of macrophages with CHI3L1 leads to upregulated expression of HSC-activation factors, suggesting its role in modulating macrophage-HSC crosstalk.
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Quitinases , Subunidade alfa2 de Receptor de Interleucina-13 , Hepatopatia Gordurosa não Alcoólica , Camundongos , Humanos , Animais , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/metabolismo , Quitinases/metabolismo , Subunidade alfa2 de Receptor de Interleucina-13/metabolismo , Fígado/metabolismo , Cirrose Hepática/metabolismo , Dieta Hiperlipídica/efeitos adversos , Camundongos Knockout , Camundongos Endogâmicos C57BL , Modelos Animais de DoençasRESUMO
Due to the higher birth rate of preterm infants and improvements in their management, metabolic bone disease of prematurity (MBDP) has a high incidence and is attracting attention. However, clear indicators for the early diagnosis of MBDP are lacking. We aimed to explore simple and feasible early warning indicators for diagnosing MBDP. Our study collected case data of premature infants from two medical centers in Chongqing from January 2020 to February 2022. According to the inclusion and exclusion criteria, data from 136 cases were collected. The correlation between 14 variables in each case and the occurrence of MBDP was analyzed. According to area under the receiver operating characteristic curve (AUROC) analysis, the best cutoff value for each variable was determined. Potential predictors were selected, and Least Absolute Shrinkage and Selection Operator (LASSO) regression analysis was used to establish the association of two models with MBDP, whose results were used to develop a diagnostic nomogram. Furthermore, a model decision curve was analyzed. Four predictors were selected from 14 clinical variables by LASSO regression, and Model I was established, including the following characteristics: height (>36â¯cm), head circumference (≤29.49â¯cm), total serum calcium (Ca) (>2.13â¯mmol/L), and alkaline phosphatase (ALP) (>344â¯U/L) levels. A single predictor, the ALP level (>344â¯U/L), was used to establish Model II. The AUROC values of the two models were 0.959 for Model I and 0.929 for Model II. In conclusion, in this study, two diagnostic models of MBDP were developed using four combinations of predictors and ALP as a single predictor. Both models showed good sensitivity and specificity for the early diagnosis of metabolic bone disease (MBD), and an ALP level of 344â¯U/L was defined as a simple and effective diagnostic threshold. In future studies, using larger samples, diagnostic threshold values of ALP for premature infants of different ages should be established, and internal and external validations are needed to improve the adaptability of the current model.
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Doenças Ósseas Metabólicas , Recém-Nascido Prematuro , Humanos , Recém-Nascido , Doenças Ósseas Metabólicas/metabolismo , Cálcio , Curva ROC , Sensibilidade e EspecificidadeRESUMO
STAT6 (signal transducer and activator of transcription 6) is a transcription factor that plays a central role in the pathophysiology of allergic inflammation. We have identified 16 patients from 10 families spanning three continents with a profound phenotype of early-life onset allergic immune dysregulation, widespread treatment-resistant atopic dermatitis, hypereosinophilia with esosinophilic gastrointestinal disease, asthma, elevated serum IgE, IgE-mediated food allergies, and anaphylaxis. The cases were either sporadic (seven kindreds) or followed an autosomal dominant inheritance pattern (three kindreds). All patients carried monoallelic rare variants in STAT6 and functional studies established their gain-of-function (GOF) phenotype with sustained STAT6 phosphorylation, increased STAT6 target gene expression, and TH2 skewing. Precision treatment with the anti-IL-4Rα antibody, dupilumab, was highly effective improving both clinical manifestations and immunological biomarkers. This study identifies heterozygous GOF variants in STAT6 as a novel autosomal dominant allergic disorder. We anticipate that our discovery of multiple kindreds with germline STAT6 GOF variants will facilitate the recognition of more affected individuals and the full definition of this new primary atopic disorder.
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Asma , Hipersensibilidade Alimentar , Humanos , Fator de Transcrição STAT6 , Mutação com Ganho de Função , Imunoglobulina E/genéticaRESUMO
The neuromedin B receptor (Nmbr) is an important physiological regulator of spontaneous activities and stress responses through different cascades as well as its autocrine and paracrine effects. Previous studies have revealed that neuromedin B (Nmb) and its receptor signal via the Rela (also known as p65)/Il6 pathway in a mouse model of pregnancy. This study investigated the mechanism of Nmbr signaling via the Rela/p65-Il6 pathway and regulation of the concentration of intracellular free calcium ([Ca(2+)](i)) during the onset of labor in primary mouse myometrial cell cultures isolated from mice in term labor. Data demonstrated Nmbr agonist-mediated upregulation of the DNA binding activity of Rela/p65, Il6 expression, and [Ca(2+)](i) in a concentration-dependent manner. Furthermore, a significant correlation was observed between DNA binding activity of Rela/p65 and Il6 expression. Moreover, this up-regulation was blocked by Nmbr and Rela/p65 knockdown, achieved by RNA interference (RNAi) technology. No significant differences were identified in the inhibition of Il6 expression as a result of Nmbr or Rela/p65 knockdown. However, significant differences were observed between the [Ca(2+)](i) in Rela/p65-specific group and that in the Nmbr-specific small interfering RNA (siRNA)-treated groups. These data demonstrated that the Nmb/Nmbr interaction in pregnant myometrial primary cells in vitro predominantly influenced uterine activity through regulation of Il6 expression via the Rela/p65 pathway, although the effects of Nmbr on [Ca(2+)](i) involved several pathways that remain to be elucidated.
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Interleucina-6/biossíntese , Miométrio/fisiologia , Neurocinina B/análogos & derivados , Receptores da Bombesina/fisiologia , Fator de Transcrição RelA/fisiologia , Animais , Cálcio/fisiologia , Células Cultivadas , Feminino , Início do Trabalho de Parto/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Neurocinina B/fisiologia , Gravidez , Interferência de RNA/fisiologia , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
BACKGROUND: Chinese primary care settings have a heavy patient load, shortage of physicians, limited medical resources and low medical literacy, making it difficult to screen for developmental disorders in infants. The Infant Neurological International Battery (INFANIB) for the assessment of neuromotor developmental disorders in infants aged 0 ~ 18 months is widely applied in community health service centers because of its simplicity, time-saving advantages and short learning curve. We aimed to develop and assess a Chinese version of the INFANIB. METHODS: A Chinese version of the INFANIB was developed. Fifty-five preterm and 49 full-term infants with high risk of neurodevelopmental delays were assessed using the Chinese version of the INFANIB at 3, 7 and 10 months after birth. The Peabody Developmental Motor Scale (PDMS) was simultaneously used to assess the children with abnormalities and diagnose cerebral palsy. The sensitivity, specificity, positive predictive value and negative predictive value of the scale were calculated. RESULTS: At birth, a higher proportion of full-term infants had asphyxia (p < 0.001), brain damage ( p = 0.003) and hyperbilirubinemia ( p = 0.022). The interclass correlation coefficient and intraclass correlation coefficient values for the INFANIB at 3, 7 and 10 months were >0.8, indicating excellent reliability with regard to inter- and intraobserver differences. The specificity, sensitivity, positive predictive value and negative predictive value were high for both high-risk premature infants and full-term infants at the age of 10 months. For premature infants at the age of 7 months or below, INFANIB had low validity for detecting abnormalities. CONCLUSIONS: The Chinese version of the INFANIB can be useful for screening infants with high-risk for neuromotor abnormality in Chinese primary care settings.
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Paralisia Cerebral/diagnóstico , Deficiências do Desenvolvimento/diagnóstico , Testes Neuropsicológicos , Transtornos Psicomotores/diagnóstico , China , Estudos de Viabilidade , Seguimentos , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Doenças do Prematuro/diagnóstico , Variações Dependentes do Observador , Valor Preditivo dos Testes , Atenção Primária à Saúde , Reprodutibilidade dos Testes , Risco , Sensibilidade e EspecificidadeRESUMO
BACKGROUND & AIMS: Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease worldwide. The NLRP3 inflammasome, a platform for caspase-1 activation and release of interleukin 1ß, is increasingly recognized in the induction of inflammation and liver fibrosis during NAFLD. However, the cell-specific contribution of NLRP3 inflammasome activation in NAFLD remains unknown. METHODS: To investigate the role of NLRP3 inflammasome activation in hepatocytes, hepatic stellate cells (HSCs) and myeloid cells, a conditional Nlrp3 knock-out mouse was generated and bred to cell-specific Cre mice. Both acute and chronic liver injury models were used: lipopolysaccharide/adenosine-triphosphate to induce in vivo NLRP3 activation, choline-deficient, L-amino acid-defined high-fat diet, and Western-type diet to induce fibrotic nonalcoholic steatohepatitis (NASH). In vitro co-culture studies were performed to dissect the crosstalk between myeloid cells and HSCs. RESULTS: Myeloid-specific deletion of Nlrp3 blunted the systemic and hepatic increase in interleukin 1ß induced by lipopolysaccharide/adenosine-triphosphate injection. In the choline-deficient, L-amino acid-defined high-fat diet model of fibrotic NASH, myeloid-specific Nlrp3 knock-out but not hepatocyte- or HSC-specific knock-out mice showed significant reduction in inflammation independent of steatosis development. Moreover, myeloid-specific Nlrp3 knock-out mice showed ameliorated liver fibrosis and decreased HSC activation. These results were validated in the Western-type diet model. In vitro co-cultured studies with human cell lines demonstrated that HSC can be activated by inflammasome stimulation in monocytes, and this effect was significantly reduced if NLRP3 was downregulated in monocytes. CONCLUSIONS: The study provides new insights in the cell-specific role of NLRP3 in liver inflammation and fibrosis. NLRP3 inflammasome activation in myeloid cells was identified as crucial for the progression of NAFLD to fibrotic NASH. These results may have implications for the development of cell-specific strategies for modulation of NLRP3 activation for treatment of fibrotic NASH.
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Inflamassomos , Cirrose Hepática , Células Mieloides , Proteína 3 que Contém Domínio de Pirina da Família NLR , Hepatopatia Gordurosa não Alcoólica , Adenosina , Aminoácidos , Animais , Caspases , Colina , Hepatite/genética , Hepatite/imunologia , Humanos , Inflamassomos/genética , Inflamassomos/imunologia , Inflamação , Interleucina-1beta/imunologia , Lipopolissacarídeos , Cirrose Hepática/genética , Cirrose Hepática/imunologia , Camundongos , Camundongos Knockout , Células Mieloides/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/imunologia , PolifosfatosRESUMO
OBJECTIVE: Alkaline phosphatase (ALP) is a ubiquitous enzyme in humans that can be used for diagnosing childhood diseases. Infants have the highest rapid growth rate and are susceptible to metabolic bone diseases. In infants, ALP activities exhibit significant month-wise variations, and authoritative standards are lacking. The present study aimed to provide a reference for the diagnosis of diseases related to abnormal ALP activities in infants. METHODS: This study included 24,618 samples collected from infants aged 0-12 months from three medical centers in Chongqing, China. Samples of infants diagnosed with diseases that may affect ALP activity have been exclude. ALP activity was analyzed using an automatic biochemical analyzer. A percentile curve for ALP activity in male and female infants was constructed using MATLAB, and the skewness-median-coefficient of variation method was employed for curve fitting. RESULTS: ALP activity in male and female infants peaked at 0-4 months; the peak appeared at 1-2 months and declined gradually thereafter. After 4-5 months of age, the ALP activities declined further, with the lowest values observed at 11-12 months of age. A comparison between the data from this study and a those from a published German study indicates that Chinese infants exhibited peak ALP activity later and subsequent decline greater than German infants. CONCLUSIONS: A percentile curve was constructed for month-wise ALP activity in male and female infants, which could provide a reference for diagnosing diseases related to abnormal ALP activity in infants.
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Fosfatase Alcalina/sangue , Doenças Ósseas Metabólicas/sangue , Doenças do Recém-Nascido/sangue , China , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Fatores SexuaisRESUMO
Talaromyce marneffei is an important thermally dimorphic pathogen causing disseminated mycoses in immunocompromised individuals in southeast Asia. Previous studies have suggested that NLRP3 inflammasome plays a critical role in antifungal immunity. However, the mechanism underlying the role of NLRP3 inflammasome activation in host defense against T. marneffei remains unclear. We show that T. marneffei yeasts but not conidia induce potent IL-1ß production. The IL-1ß response to T. marneffei yeasts is differently regulated in different cell types; T. marneffei yeasts alone are able to induce IL-1ß production in human PBMCs and monocytes, whereas LPS priming is essential for IL-1ß response to yeasts. We also find that Dectin-1/Syk signaling pathway mediates pro-IL-1ß production, and NLRP3-ASC-caspase-1 inflammasome is assembled to trigger the processing of pro-IL-1ß into IL-1ß. In vivo, mice deficient in NLRP3 or caspase-1 exhibit higher mortality rate and fungal load compared to wild-type mice after systemic T. marneffei infection, which correlates with the diminished recruitment of CD4 T cells into granulomas in knockout mice. Thus, our study first demonstrates that NLRP3 inflammasome contributes to host defense against T. marneffei infection.
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Inflamassomos/imunologia , Micoses/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Infecções Oportunistas/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Caspase 1/genética , Feminino , Humanos , Inflamassomos/genética , Interleucina-1beta/imunologia , Lectinas Tipo C/imunologia , Leucócitos Mononucleares/imunologia , Fígado/imunologia , Fígado/microbiologia , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Micoses/microbiologia , Micoses/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Infecções Oportunistas/microbiologia , Infecções Oportunistas/patologia , Baço/microbiologia , TalaromycesRESUMO
BACKGROUND: This study aimed to evaluate the correlation and consistency between traditional head measurement and structured light three-dimensional (3D) scanning parameters when measuring infant skull shape. METHODS: A total of 76 infants aged 3 months to 2.5 years old were included in the study. Head circumference (HC) was measured with a tape measure. The transverse, anteroposterior, and oblique diameters were measured using a spreading caliper, and the cranial vault asymmetry index (CVAI) and a cranial index (CI) of symmetry were calculated; 76 cases were measured successfully. The above indexes were measured using a structured light 3D scanning system (71 cases were measured with success). Thus, in the end, the valid data of 71 cases were analyzed, and the measurements of the two approaches were compared. RESULTS: The 95% confidence interval of traditional head measurement and structured light 3D scanning was between 0.633 and 0.988. Pearson's correlation coefficient indicated a high correlation between the two methods (r=0.793-0.980). The correlation coefficients of the transverse diameter, anteroposterior diameter, and HC, and the CI of symmetry were higher than 0.9. The lowest correlation coefficient for the CVAI was 0.793. The P values of the above measurement data were all <0.001, which indicated that they were closely related. A Bland-Altman plot indicated reasonable consistency between the two methods. CONCLUSIONS: Both traditional head measurement and structured light 3D scanning are suitable for the measurement of infant head shape. However, while traditional head measurement using a spreading caliper is economical and simple, making it suitable for general screening at a basic level, structured light 3D scanning can deliver additional parameters, which is useful for infants with an abnormal head shape. The latter is also convenient for designing a customized helmet for skull correction when needed.
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AIMS: Preeclampsia (PE) accounts for the foremost cause of maternal and fetal mortality worldwide, whereas, there are no effective treatments for the disease yet. Long non-coding RNAs (lncRNAs) play critical roles in various human disorders, including PE. Here, we identified an up-regulated lncRNA HOTAIR, and explored its underlying mechanisms in PE. MAIN METHODS: qRT-PCR analysis was used to examine HOTAIR expression in PE tissues and cell lines. Trophoblast proliferation was examined by colony formation and 5-Ethynyl-2'-deoxyuridine (EdU) incorporation assays. Trophoblast migration and invasion was determined by transwell and wound healing assays. Bioinformatics analysis was performed to verify the regulation HOTAIR on miRNAs. The interaction between HOTAIR and EZH2 was detected using RNA immunoprecipitation assay (RIP). Chromatin immunoprecipitation (CHIP) assay was also performed to verify that the negative regulation of HOTAIR on miR-106a was dependent on the epigenetic repressor EZH2. KEY FINDINGS: HOTAIR was up-regulated in PE placenta tissues, which repressed the proliferation, migration and invasion of trophoblast cells. HOTAIR significantly repressed miR-106a expression and the reduced miR-106a level was also observed in placentas from PE patients. Additionally, miR-106a mimic enhanced the migration and invasion of trophoblast cells. Further mechanistic analyses implied that the action of HOTAIR is moderately attributable to its repression of miR-106a via association with EZH2. SIGNIFICANCE: High level of HOTAIR repressed the proliferation, migration and invasion of trophoblast cells through targeting miR-106 in an EZH2-dependent manner, which may provide new insights into the roles of HOTAIR and miR-106a as potential regulators in PE.
Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , MicroRNAs/metabolismo , Pré-Eclâmpsia/metabolismo , RNA Longo não Codificante/metabolismo , Sequência de Bases , Linhagem Celular , Proliferação de Células , Progressão da Doença , Repressão Epigenética , Feminino , Humanos , Placenta/metabolismo , Gravidez , Trofoblastos/citologia , Regulação para CimaRESUMO
MicroRNA21 (miR21) is reported to exhibit cancerpromoting activity in various types of cancer. It has been previously demonstrated that miR21 is overexpressed in bladder tumor tissue compared with normal mucosa. However, the functional mechanism of miR21 in bladder cancer remains largely unknown. Thus, the current study aimed to determine the roles of miR21 in autophagy and the malignant development of bladder cancer in T24 cells. Upregulation or downregulation of miR21 was achieved following the transfection of miR21 mimic or miR21 inhibitor. An MTT assay was additionally performed to measure cell growth. Wound healing and transwell invasion assays were used to detect cell migration and invasion. The apoptotic potential and cell cycle were examined via flow cytometry and reverse transcriptionquantitative PCR was performed to evaluate the expression of phosphatase and tensin homolog (PTEN), beclin 1, microtubuleassociated protein l light chain 3B (LC3II), cyclin D1, caspase3, Ecadherin, matrix metallopeptidase9 (MMP9) and vimentin. The results revealed that the proliferation, migration and invasion of T24 cells was greatly increased in the miR21 mimic group, while apoptosis was greatly inhibited. Additionally, T24 cells treated with miR21 mimic exhibited G1phase arrest. In the miR21 mimic group, the expression of PTEN, beclin 1, LC3II, caspase3 and Ecadherin were decreased, while the expression of cyclin D1, MMP9 and vimentin were increased. Opposite effects were observed in the miR21 inhibitor group. The data of the current study may indicate that miR21 overexpression inhibited autophagy and promoted the proliferation, migration, invasion and epithelial to mesenchymal transition of bladder cancer T24 cells. The results may further elucidate the molecular mechanism of miR21 in the development of bladder cancer.
Assuntos
Autofagia , Transição Epitelial-Mesenquimal , MicroRNAs/genética , Neoplasias da Bexiga Urinária/patologia , Apoptose , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , MicroRNAs/metabolismo , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismoRESUMO
PDB-1 is a new C-27-carboxylated-lupane-triterpenoid derivative isolated from Potentilla discolor Bunge. In our previous research, PDB-1 was suggested to have an obvious selectivity for tumor cells. This study focused on clarifying PDB-1's anticancer mechanism in the inhibition of proliferation and in the induction of apoptosis and autophagy in A549 cells. In general, A549 cells were treated with PDB-1 for different times, and cell survival was assessed by a CCK8 assay. The assessment of intracellular reactive oxygen species, a mitochondrial membrane potential assay, a cell cycle assay, an annexin V-FITC/PI assay, and MDC staining were performed in A549 cells treated with PDB-1. Moreover, the mRNA and protein expression of cell cycle-, apoptosis- and autophagy-related factors were detected by RT-qPCR and western blotting. The results showed that PDB-1 inhibited A549 cell proliferation and colony formation in a dose- and time-dependent manner. The decrease in the viability of A549 cells was due to a G2/M cell cycle arrest. Moreover, PDB-1 induced cell apoptosis, accompanied by an increase in the Bax/Bcl-2 ratio and an increase in the expression levels of cleaved caspase-3/caspase-9. We also found that PDB-1 induced autophagy by increasing the conversion of LC3-I to LC3-II and elevating Beclin-1. In addition, further studies indicated that pretreatment with a specific PI3K inhibitor (LY294002) enhanced the effects of PDB-1 on the expression of proteins associated with apoptosis and autophagy, demonstrating that the PI3K/Akt/mTOR pathway was related to PDB-1-induced apoptosis and autophagy. These results indicated that PDB-1 may be considered a potential candidate for the future treatment of lung adenocarcinoma. These findings should benefit the development of the C14-COOH type of pentacyclic triterpenoids.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias Pulmonares/tratamento farmacológico , Fosfatidilinositol 3-Quinase/metabolismo , Extratos Vegetais/farmacologia , Potentilla , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Triterpenos/farmacologia , Células A549 , Antineoplásicos Fitogênicos/isolamento & purificação , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proteínas Relacionadas à Autofagia/genética , Proteínas Relacionadas à Autofagia/metabolismo , Proliferação de Células/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Células HeLa , Células Hep G2 , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Células MCF-7 , Extratos Vegetais/isolamento & purificação , Potentilla/química , Transdução de Sinais , Triterpenos/isolamento & purificaçãoRESUMO
OBJECTIVE: Alkaline phosphatase (ALP) serves as a biomarker for diagnosing several types of diseases in adults; nonetheless, its use is restricted in children because of changes in ALP activity during different physiological phases. The present study aimed to investigate ALP activity and its dynamics in children of different ages to establish the reference values for ALP activity in children. METHODS: Total 167,625 samples of children aged 0-18 years were enrolled in this study. ALP activity was measured using the 4-nitrophenyl-1-phosphate disodium salt (4-NPP)-2-amino-2-methyl-1-propanol (AMP) method with an automatic biochemical analyzer. Patients with known diagnoses that may affect ALP activity were excluded. A percentile curve was plotted using MATLAB software, and the curve was fitted using the skewness-median-coefficient of variation (LMS) method. RESULTS: ALP activity reached the highest peak at 12-13 years of age and then gradually decreased to the lowest peak at 18-19 years of age in boys, whereas it reached the highest at 10-11 years and then gradually reduced to the lowest at 17-18 years in girls. Furthermore, the highest peak of ALP activity appeared substantially earlier in children of either sex in China than in those in Germany. CONCLUSIONS: We showed the dynamics of ALP activity in both boys and girls between the ages of 0 and 18 years in China and compared the difference in ALP activity between children in China and Germany. Our findings provide a reference for clinicians.
Assuntos
Desenvolvimento do Adolescente/fisiologia , Fosfatase Alcalina/sangue , Desenvolvimento Infantil/fisiologia , Adolescente , Criança , Pré-Escolar , China , Ativação Enzimática , Feminino , Alemanha , Humanos , Lactente , Recém-Nascido , Masculino , Valores de Referência , Adulto JovemRESUMO
Talaromyces (Penicillium) marneffei is an AIDS-defining infection in Southeast Asia and is associated with high mortality. It is rare in non-immunosuppressed individuals, especially children. Little is known about host immune response and genetic susceptibility to this endemic fungus. Genetic defects in the interferon-gamma (IFN-γ)/STAT1 signaling pathway, CD40/CD40 ligand- and IL12/IL12-receptor-mediated crosstalk between phagocytes and T-cells, and STAT3-mediated Th17 differentiation have been reported in HIV-negative children with talaromycosis and other endemic mycoses such as histoplasmosis, coccidioidomycosis, and paracoccidioidomycosis. There is a need to design a diagnostic algorithm to evaluate such patients. In this article, we review a cohort of pediatric patients with disseminated talaromycosis referred to the Asian Primary Immunodeficiency Network for genetic diagnosis of PID. Using these illustrative cases, we propose a diagnostics pipeline that begins with immunoglobulin pattern (IgG, IgA, IgM, and IgE) and enumeration of lymphocyte subpopulations (T-, B-, and NK-cells). The former could provide clues for hyper-IgM syndrome and hyper-IgE syndrome. Flow cytometric evaluation of CD40L expression should be performed for patients suspected to have X-linked hyper-IgM syndrome. Defects in interferon-mediated JAK-STAT signaling are evaluated by STAT1 phosphorylation studies by flow cytometry. STAT1 hyperphosphorylation in response to IFN-α or IFN-γ and delayed dephosphorylation is diagnostic for gain-of-function STAT1 disorder, while absent STAT1 phosphorylation in response to IFN-γ but normal response to IFN-α is suggestive of IFN-γ receptor deficiency. This simple and rapid diagnostic algorithm will be useful in guiding genetic studies for patients with disseminated talaromycosis requiring immunological investigations.