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Tuberculous myocarditis (TM) is an extremely rare manifestation of Mycobacterium tuberculosis (TB) infection. Although TM is a critical cause of sudden cardiac death, only a few cases have been reported. We report the case of an older patient with pulmonary TB with a history of fever, chest tightness, paroxysmal palpitations, and electrocardiographic evidence of sinus node conduction abnormalities on admission. Although emergency physicians observed these unusual clinical manifestations, no timely differential diagnosis was made nor interventions were performed. A definitive diagnosis of TM and histopathological findings compatible with sinus node involvement were made based on autopsy outcomes. Herein, we describe the clinical presentation and pathological features of a rare form of Mycobacterium TB. In addition, we provide an overview of issues related to the diagnosis of myocardial TB.
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Cadmium is a naturally existing heavy metal and a notorious environmental pollutant. While its toxic outcomes and underlying mechanisms remain largely elusive. To explore the behavioral change caused by multigenerational exposure of cadmium to C. elegans, we challenged the C. elegans with cadmium for six generations and observed its impact on animal behaviors. Wild-type worms were randomly divided into two groups, the control and cadmium exposure groups. Locomotive and chemotactic behaviors were observed across six generations. Head thrashing frequency, chemotaxis index, and fold change index were used to evaluate the neurotoxicity of multigenerational cadmium exposure. Multigenerational cadmium exposure can trans-generationally increase the head thrashing frequency of C. elegans during swimming, and impair the chemotactic behaviors to isoamyl alcohol, diacetyl, and 2-nonanone. Our findings proposed a trans-generationally behavioral impact induced by multigenerational cadmium exposure.
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Caenorhabditis elegans , Metales Pesados , Animales , Conducta Animal , Cadmio/toxicidadRESUMEN
We assessed the effectiveness of mRNA and viral-vector vaccines in epidemic period led by different SARS-CoV-2 variants. Systematic search of PubMed, EMBASE, and CNKI (China National Knowledge Infrastructure) databases without language restriction for studies published before September 19, 2022. The review was registered with PROSPERO (CRD42022335430) and reported according to PRISMA guidelines. Forty studies met the inclusion criteria for this study, with 62 954 861 participants. The overall vaccine effectiveness (VE) to prevent COVID-19 infection was 0.76 (95% confidence interval [CI] 0.73-0.78), symptomatic infection was 0.87 (95% CI 0.83-0.91), hospital admissions was 0.82 (95% CI 0.75-0.87), and mortality was 0.76 (95% CI 0.48-0.89). Subgroup analysis were performed to characterize the effectiveness of different vaccines. When SARS-CoV-2 variants are taking account, the VE decreased along with the variation of the virus by clinical outcomes and vaccine types. The findings of this systematic review provide the best available evidence that BNT162b2, mRNA-1273, ChAdOx1, and Ad26. COV2.S seems to be approximately effective from predelta to omicron, but only modestly effective in participants aged 65 or older. When SARS-CoV-2 variants are taking account, VE decreased along with the variation of the virus for all mRNA and viral-vector vaccines.
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COVID-19 , Vacunas Virales , Humanos , SARS-CoV-2/genética , ARN Mensajero , Vacuna BNT162 , COVID-19/epidemiología , COVID-19/prevención & controlRESUMEN
Objective: Amphetamine-type stimulant (ATS) and opioid dependencies are chronic inflammatory diseases with similar symptoms and common genomics. However, their coexpressive genes have not been thoroughly investigated. We aimed to identify and verify the coexpressive hub genes and pathway involved in the pathogenesis of ATS and opioid dependencies. Methods: The microarray of ATS- and opioid-treatment mouse models was obtained from the Gene Expression Omnibus database. GEO2R and Venn diagram were performed to identify differentially expressed genes (DEGs) and coexpressive DEGs (CDEGs). Functional annotation and protein-protein interaction network detected the potential functions. The hub genes were screened using the CytoHubba and MCODE plugin with different algorithms, and further validated by receiver operating characteristic analysis in the GSE15774 database. We also validated the hub genes mRNA levels in BV2 cells using qPCR. Result: Forty-four CDEGs were identified between ATS and opioid databases, which were prominently enriched in the PI3K/Akt signaling pathway. The top 10 hub genes were mainly enriched in apoptotic process (CD44, Dusp1, Sgk1, and Hspa1b), neuron differentiation, migration, and proliferation (Nr4a2 and Ddit4), response to external stimulation (Fos and Cdkn1a), and transcriptional regulation (Nr4a2 and Npas4). Receiver operating characteristic (ROC) analysis found that six hub genes (Fos, Dusp1, Sgk1, Ddit4, Cdkn1a, and Nr4a2) have an area under the curve (AUC) of more than 0.70 in GSE15774. The mRNA levels of Fos, Dusp1, Sgk1, Ddit4, Cdkn1a, PI3K, and Akt in BV2 cells and GSE15774 with METH and heroin treatments were higher than those of controls. However, the Nr4a2 mRNA levels increased in BV2 cells and decreased in the bioinformatic analysis. Conclusions: The identification of hub genes was associated with ATS and opioid dependencies, which were involved in apoptosis, neuron differentiation, migration, and proliferation. The PI3K/Akt signaling pathway might play a critical role in the pathogenesis of substance dependence.
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Hepatitis B virus (HBV) X protein (HBx) has been determined to play a crucial role in the replication and transcription of HBV, and its biological functions mainly depend on the interaction with other host proteins. This study aims at screening the proteins that bind to the key functional domain of HBx by integrated proteomics. Proteins that specifically bind to the transactivation domain of HBx were selected by comparing interactors of full-length HBx and HBx-D5 truncation determined by glutathione-S-transferase (GST) pull-down assay combined with mass spectrometry (MS). The function of HBx interactor Pin1 in HBV replication was further investigated by in vitro experiments. In this study, a total of 189 proteins were identified from HepG2 cells that specifically bind to the transactivation domain of HBx by GST pull-down and subsequent MS. After gene ontology (GO) analysis, Pin1 was selected as the protein with the highest score in the largest cluster functioning in protein binding, and also classified into the cluster of proteins with the function of structural molecule activity, which is of great potential to be involved in HBV life cycle. The interaction between Pin1 and HBx has been further confirmed by Ni2+-NTA pulldown assay, co-immunoprecipitation, and immunofluorescence microscopy. HBsAg and HBeAg levels significantly decreased in Pin1 expression inhibited HepG2.2.15 cells. Besides, the inhibition of Pin1 expression in HepG2 cells impeded the restored replication of HBx-deficient HBV repaired by ectopic HBx expression. In conclusion, our study identified Pin1 as an interactor binds to the transactivation domain of HBx, and suggested the potential association between Pin1 and the function of HBx in HBV replication.
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Virus de la Hepatitis B/fisiología , Peptidilprolil Isomerasa de Interacción con NIMA/metabolismo , Transactivadores/química , Transactivadores/metabolismo , Activación Transcripcional , Proteínas Reguladoras y Accesorias Virales/química , Proteínas Reguladoras y Accesorias Virales/metabolismo , Replicación Viral/fisiología , Células Hep G2 , Humanos , Unión Proteica , Dominios Proteicos , Mapas de Interacción de Proteínas , Vía de Señalización WntRESUMEN
The eyelid folding represents one of the most distinguishing features of East Asian faces, involving the absence or presence of the eyelid crease, i.e., single vs. double eyelid. Recently, a genome-wide association study (GWAS) identified two SNPs (rs12570134 and rs1415425) showing genome-wide significant association with the double eyelid phenotype in Japanese. Here we report a confirmatory study in 697 Chinese individuals of exclusively Han origin. Only rs1415425 was statistically significant (P-value = 0.011), and the allele effect was on the same direction with that reported in Japanese. This SNP combined with gender and age explained 10.0% of the total variation in eyelid folding. DNA-based prediction model for the eyelid trait was developed and evaluated using logistic regression. The model showed mild to moderate predictive capacity (AUC = 0.69, sensitivity = 63%, and specificity = 70%). We further selected six additional SNPs by massive parallel sequencing of 19 candidate genes in 24 samples, and one SNP rs2761882 was statistically significant (P-value = 0.027). All predictors including these two SNPs (rs1415425 and rs2761882), gender, and age explained 11.2% of the total variation. The combined prediction model obtained an improved predictive capacity (AUC = 0.72, sensitivity = 62%, and specificity = 66%). Our study thus provided a confirmation of previous GWAS findings and a DNA-based prediction of the eyelid trait in Chinese Han individuals. This model may add value to forensic DNA phenotyping applications considering gender and age can be separately inferred from genetic and epigenetic markers. To further improve the prediction accuracy, future studies should focus on identifying more informative SNPs by large GWASs in East Asian populations.
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Párpados/anatomía & histología , Estudio de Asociación del Genoma Completo , Secuenciación de Nucleótidos de Alto Rendimiento , Fenotipo , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN , Adulto , Área Bajo la Curva , Pueblo Asiatico/genética , China/etnología , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Sensibilidad y EspecificidadRESUMEN
INTRODUCTION: The etiology of sudden infant death syndrome (SIDS) remains an unsolved problem. The aim of this meta-analysis is to investigate the potential association between monoamine oxidase A (MAOA) promoter variable number tandem repeat (VNTR) polymorphism and SIDS risk. METHODS: A systematic review and meta-analysis were conducted on studies from accessible electronic databases. Each VNTR variant was examined in each gender independently by comparing with the pooled results of other alleles. RESULTS: A total of six independent case-control studies including 1022 SIDS cases and 1839 controls were enrolled in this meta-analysis. In both of the whole populations and Caucasian populations, male infants with the low-MAOA-expression alleles (2R+3R) were found to exhibit a statistically significant increased risk of SIDS, whereas those with a 4R allele exhibited a reduced risk of SIDS. Besides, an increased risk of SIDS was detected in male Caucasian infants with 2R or 3R alleles. However, none of the allele or genotype variants was associated with SIDS in female victims. CONCLUSION: In male Caucasian infants, the low expression of MAOA promoter VNTR alleles (2R and 3R) is associated with an increased risk of SIDS, and the existence of the 4R allele could be regarded as a protective factor.
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Alelos , Repeticiones de Minisatélite , Monoaminooxidasa/genética , Polimorfismo Genético , Regiones Promotoras Genéticas , Muerte Súbita del Lactante/genética , Estudios de Casos y Controles , Femenino , Genotipo , Humanos , Lactante , Masculino , Población Blanca/genéticaRESUMEN
The developmental patterns of the pelvic epiphyses are one of the anatomical markers used in the assessment of skeletal age and the legally relevant age threshold. In this study, four regression models and five classification models were developed for forensic age estimation and the determination of the 18-year threshold, respectively. A total of 2137 conventional pelvic radiographs (1215 males and 922 females) aged 10.00-25.99 years were analyzed, and the ossification and fusion of the iliac crest and ischial tuberosity epiphyses were scored separately. The epiphyses on both sides were used as inputs for all models. The accuracy of the regression models was compared using the mean absolute error (MAE) and root mean square error. The percentages of correct classifications were evaluated for the determination of the 18-year threshold. Support vector regression (SVR) and gradient boosting regression (GBR) showed higher accuracy for age estimation in both sexes. The lowest MAE was 1.38 years in males when using SVR and 1.16 years in females when using GBR. In the demarcation of minors and adults, the percentage of correct classification was over 92%, and the area under the receiver operating characteristic curves was over 0.91 in all models, except the Bernoulli naive Bayes classifier. This study demonstrated that the present models may be helpful for age estimation and the determination of the 18-year threshold. However, owing to the high effective dose of ionizing radiation used during conventional radiography of the pelvis, it is expected that these models will be tested with pelvic MRI for age estimation.
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Determinación de la Edad por el Esqueleto/métodos , Ilion/diagnóstico por imagen , Isquion/diagnóstico por imagen , Modelos Estadísticos , Adolescente , Adulto , Pueblo Asiatico , Niño , China , Epífisis/anatomía & histología , Epífisis/diagnóstico por imagen , Etnicidad , Femenino , Antropología Forense , Humanos , Ilion/anatomía & histología , Isquion/anatomía & histología , Masculino , Osteogénesis , Radiografía , Máquina de Vectores de Soporte , Adulto JovenRESUMEN
OBJECTIVES: This study aimed to explore whether computed tomography (CT) images of cranial sutures can contribute to adult age estimation in Chinese Han individuals. MATERIALS AND METHODS: This study was based on cranial CT scans of 230 Chinese Han males aged 23.33-76.93 years. A total of 160 images from 16 suture segments were scored after volume reformation and multiplanar reconstruction in each individual. Decision tree regression, linear support vector regression, Bayesian ridge regression, and gradient boosting regression were developed for adult age estimation by a training set using leave-one-out cross-validation and further evaluated by the test set. The inaccuracy and bias were calculated to evaluate the four models and the previously used models from the literature. RESULTS: The degree of suture closure was associated with adult age. The minimum inaccuracy of the test set was 7.73 years obtained by linear support vector regression, while the inaccuracy of previous simple linear regression models was 13.09 and 10.97 years. The accuracy was higher in the age group from 40.00 to 59.99 years compared to the other age groups. DISCUSSION: The accuracy of our models for adult age estimation was superior to those in previous studies based on cranial sutures. Hence, the application of novel statistical data mining tools helps to improve aging issues. Nevertheless, age estimation of adults should be combined with other methods, since the accuracy level is still not satisfactory.
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Suturas Craneales/anatomía & histología , Adulto , Determinación de la Edad por el Esqueleto/métodos , Anciano , Teorema de Bayes , China , Árboles de Decisión , Humanos , Modelos Lineales , Masculino , Persona de Mediana Edad , Tomografía Computarizada Multidetector , Adulto JovenRESUMEN
Hepatitis B virus X protein (HBx) can stimulate the transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-determining enzyme in gluconeogenic pathway. Two isoforms of PEPCK exist, a cytoplasmic form (PCK1) and a mitochondrial isoform (PCK2). The current study investigated the direct effect of HBx-stimulated PEPCK on hepatitis B virus (HBV) replication. We showed that PCK1 rather than PCK2 was upregulated by HBx. We also demonstrated that overexpression of PCK1 decreased HBV replication, whereas inhibition of PCK1-enhanced HBV replication. Furthermore, we found overexpression of PCK1 led to reduced expression of peroxisome proliferator-activated receptor-coactivator 1α (PGC-1α) and peroxisome proliferator-activated receptor γ (PPAR-γ), whereas knocking down PCK1 resulted in an increased expression of PGC-1α and PPAR-γ. When PPAR-γ was inhibited, knocking down PCK1 could not induce the apparent enhanced HBV replication. Our data suggested that PCK1 induced by HBx led to decreased HBV replication through the downregulation of PGC-1α and PPAR-γ. Thus, our study demonstrates a negative-feedback loop involving PCK1 and HBV may provide a balanced cell environment for HBV persistent infection.
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Expresión Génica , Virus de la Hepatitis B/crecimiento & desarrollo , Interacciones Huésped-Patógeno , Péptidos y Proteínas de Señalización Intracelular/biosíntesis , Fosfoenolpiruvato Carboxiquinasa (GTP)/biosíntesis , Transactivadores/metabolismo , Replicación Viral , Línea Celular , Hepatocitos/virología , Humanos , PPAR gamma/biosíntesis , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/biosíntesis , Transcripción Genética , Proteínas Reguladoras y Accesorias ViralesRESUMEN
Spinal cord injury (SCI) is a severe traumatic lesion of central nervous system (CNS) with only a limited number of restorative therapeutic options. Diosgenin glucoside (DG), a major bioactive ingredient of Trillium tschonoskii Max., possesses neuroprotective effects through its antioxidant and anti-apoptotic functions. In this study, we investigated the therapeutic benefit and underlying mechanisms of DG treatment in SCI. We found that in Sprague-Dawley rats with traumatic SCI, the expressions of autophagy marker Light Chain 3 (LC3) and Beclin1 were decreased with concomitant accumulation of autophagy substrate protein p62 and ubiquitinated proteins, indicating an impaired autophagic activity. DG treatment, however, significantly attenuated p62 expression and upregulated the Rheb/mTOR signaling pathway (evidenced as Ras homolog enriched in brain) due to the downregulation of miR-155-3p. We also observed significantly less tissue injury and edema in the DG-treated group, leading to appreciable functional recovery compared to that of the control group. Overall, the observed neuroprotection afforded by DG treatment warrants further investigation on its therapeutic potential in SCI.
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Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Diosgenina/análogos & derivados , Glucósidos/uso terapéutico , Fármacos Neuroprotectores/uso terapéutico , Traumatismos de la Médula Espinal/prevención & control , Animales , Diosgenina/química , Diosgenina/uso terapéutico , Glucósidos/química , MicroARNs/genética , Fármacos Neuroprotectores/química , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Médula Espinal/patología , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/patología , Trillium/químicaRESUMEN
Mitochondria are an essential component of multicellular life and play important roles in the health of the cells and the body. Mitochondria can produce energy by oxidative phosphorylation, mediate calcium and reactive oxygen signal transduction, and regulate cell apoptosis. Recent studies indicate that mitochondria continually change their shapes and distribution by fission and fusion, which are collectively termed mitochondrial dynamics. Mitochondrial dynamics play critical roles in maintaining mitochondrial function. This review focuses on the structure and biological functions of mitochondrial fission and fusion related proteins in mammal cells.
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Mitocondrias/fisiología , Dinámicas Mitocondriales , Proteínas Mitocondriales/fisiología , Animales , Apoptosis , Especies Reactivas de Oxígeno , Transducción de SeñalRESUMEN
PURPOSE: The aim of this study was to screen for novel host proteins that play a role in HBx augmenting Hepatitis B virus (HBV) replication. EXPERIMENTAL DESIGN: Three HepG2 cell lines stably harboring different functional domains of HBx (HBx, HBx-Cm6, and HBx-Cm16) were cultured. ITRAQ technology integrated with LC-MS/MS analysis was applied to identify the proteome differences among these three cell lines. RESULTS: In brief, a total of 70 different proteins were identified among HepG2-HBx, HepG2-HBx-Cm6, and HepG2-HBx-Cm16 by double repetition. Several differentially expressed proteins, including p90 ribosomal S6 kinase 2 (RSK2), were further validated. RSK2 was expressed at higher levels in HepG2-HBx and HepG2-HBx-Cm6 compared with HepG2-HBx-Cm16. Furthermore, levels of HBV replication intermediates were decreased after silencing RSK2 in HepG2.2.15. An HBx-minus HBV mutant genome led to decreased levels of HBV replication intermediates and these decreases were restored to levels similar to wild-type HBV by transient ectopic expression of HBx. After silencing RSK2 expression, the levels of HBV replication intermediates synthesized from the HBx-minus HBV mutant genome were not restored to levels that were observed with wild-type HBV by transient HBx expression. CONCLUSION AND CLINICAL RELEVANCE: Based on iTRAQ quantitative comparative proteomics, RSK2 was identified as a novel host protein that plays a role in HBx augmenting HBV replication.
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Virus de la Hepatitis B/fisiología , Proteómica , Proteínas Quinasas S6 Ribosómicas 90-kDa/metabolismo , Replicación Viral , Células Hep G2 , Virus de la Hepatitis B/metabolismo , Humanos , Dominios Proteicos , Proteínas Quinasas S6 Ribosómicas 90-kDa/químicaRESUMEN
Hepatitis B virus (HBV) infection is one of the major health problems in the world. Transgelin-2 (TAGLN2) expression has been revealed to be significantly altered in previous studies concerning HBV-host interaction. The present study investigated TAGLN2 expression patterns in HBV related hepatocellular carcinoma (HCC) tissues and its role in HBV transcription and replication. We collected 59 HBV related HCC tissue samples, their adjacent non-tumoral tissues and 16 normal livers to make the tissue microarray. TAGLN2 protein was detected by immunohistochemistry and the transcriptional levels of TAGLN2, HBc, HBs and HBx were detected by qRT-PCR. Then we investigated the function of TAGLN2 on HBV transcription and replication in vitro by ectopic expressing or knocking down TAGLN2 in HepG2 and HepG2.2.15 cell lines. We further studied the effect of HBx on TAGLN2 expression with a Tet-on HBx expressing cell line. TAGLN2 protein expression was lower in normal livers and HBV-HCC tissues comparing to adjacent non-tumoral tissues. The transcriptional levels of TAGLN2 in HBV-HCC tissues and their adjacent tissues were positively related to that of HBc, HBs and HBx (P < 0.05). Ectopic expression of TAGLN2 in vitro could enhance HBV transcription and replication while suppressing TAGLN2 had the contrary effect. TAGLN2 could be induced by HBx in a dose-dependent manner. Our data demonstrated that TAGLN2 might be an HBx induced positive host factor involved in HBV transcription and replication and HBx related liver fibrosis and tumorigenesis.
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Carcinoma Hepatocelular/virología , Virus de la Hepatitis B/genética , Neoplasias Hepáticas/virología , Proteínas de Microfilamentos/genética , Proteínas Musculares/genética , Activación Transcripcional/genética , Replicación Viral/genética , Carcinoma Hepatocelular/metabolismo , Regulación Viral de la Expresión Génica/genética , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Proteínas Virales/genéticaRESUMEN
Whether luteolin inhibits HBV replication has not been validated and the underlying mechanism of which has never been elucidated. In this study, we show that luteolin reduces HBV DNA replication in HepG2.2.15 cells. Luteolin effectively inhibited the expression of hepatocyte nuclear factor 4α (HNF4α) and its binding to the HBV promoters in HepG2.2.15 cells. While the extracellular signal-regulated kinase (ERK) was activated by luteolin, inhibition of ERK abolished luteolin-induced HNF4α suppression. Consistently, blocking ERK attenuated the anti-HBV activity of luteolin. In a HBV replication mouse model, luteolin decreased the levels of HBsAg, HBeAg, HBV DNA replication intermediates, and the HBsAg and HBcAg expression. Taken together, our results validated the anti-HBV activity of luteolin in both in vitro and in vivo studies and established a signaling cascade consisting of ERK and HNF4α for inhibition of HBV replication by luteolin, which may be exploited for clinical application of luteolin for anti-HBV therapy.
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Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Virus de la Hepatitis B/efectos de los fármacos , Hepatitis B/tratamiento farmacológico , Factor Nuclear 4 del Hepatocito/metabolismo , Luteolina/farmacología , Replicación Viral/efectos de los fármacos , Animales , Apoptosis , Southern Blotting , Western Blotting , Proliferación Celular , Regulación hacia Abajo , Ensayo de Cambio de Movilidad Electroforética , Ensayo de Inmunoadsorción Enzimática , Quinasas MAP Reguladas por Señal Extracelular/genética , Femenino , Células Hep G2 , Hepatitis B/metabolismo , Hepatitis B/virología , Factor Nuclear 4 del Hepatocito/genética , Humanos , Técnicas para Inmunoenzimas , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
BACKGROUND: RPB5-Mediating protein (RMP) is associated with the RNA polymerase II subunit RPB5. This protein functionally counteracts the transcriptional activation of Hepatitis B Virus X protein (HBx) by competitively binding to the RPB5; however, the effects of RMP on Hepatitis B virus (HBV) transcription and replication remain unknown. OBJECTIVES: The purpose of this study was to investigate the effect of RMP on viral transcription and replication in vivo by using the hydrodynamic-based HBV replication mouse model. MATERIALS AND METHODS: Male balb/c mice were transfected with wild type (1.2 wt) or the HBx minus HBV plasmids (1.2x (-)) with or without HBx and RMP, to establish an HBV replication mouse model by hydrodynamic injection through the tail vein. The HBV RNA and HBV DNA replication intermediates (RI) were analyzed in the liver. RESULTS: RPB5-Mediating protein could inhibit HBV transcription and replication in groups transfected with the 1.2 wt and HBx. The inhibitory effect disappeared in the 1.2x (-) groups, yet it reappeared in the groups co-transfected with 1.2x (-) and HBx. An inhibitory effect was indicated at a low dose of RMP (0.3 ug, 0.5 ug and 0.7 ug) compared to the control group and groups that had received high doses of RMP. CONCLUSIONS: Our study demonstrated that a low dose of RMP could inhibit HBV transcription and replication, which is dependent on the appearance of HBx in vivo.
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The double nucleotide, A1762T and G1764A exchange (TA mutation), in the hepatitis B virus (HBV) genome basal core promoter (BCP) region is a common viral mutation in patients with chronic HBV infection. This mutation is located in the binding site of hepatocyte nuclear factor 4 (HNF4), and a number of liverenriched transcription factors are involved in the regulation of HBV transcription and replication. The aim of the present study was to investigate the biological characteristics of the HBV strain with this mutation, and the effect of HNF4 inhibition on the replication of this strain in vivo. The results indicated that in vivo the HBV strain with the TA mutation supported a higher level of pregenomic RNA transcription and HBV DNA replication, compared with the wildtype strain. Furthermore, the concentration of serum HBeAg in the TA mutant group was lower than that in the wildtype strain. Following treatment of the mice with entecavir (ETV) or tenofovir disoproxil fumarate (TDF), the transcription and replication levels of wildtype and mutant strains were reduced. In the groups treated with TDF, the inhibition effect was more marked. In hepatocytes in which HNF4 expression was specifically inhibited, the level of 3.5 kb mRNA of HBV was reduced compared with that in mouse cells with normal HNF4 expression, and HBV DNA replication levels were also reduced to a greater extent. Furthermore, following liverspecific knockdown of HNF4, the reduction in variant virus expression was greater than that of the wildtype virus. In conclusion, the replication capacity of HBV with the TA mutation was increased, and the mutation was associated with a reduction in serum HBeAg levels. This mutant strain remained sensitive to ETV and TDF, and HNF4 supported a higher replication level of TA mutant HBV in vivo.
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Virus de la Hepatitis B/genética , Hepatitis B/virología , Mutación , Animales , Antivirales/farmacología , Modelos Animales de Enfermedad , Genoma Viral , Hepatitis B/sangre , Hepatitis B/inmunología , Antígenos del Núcleo de la Hepatitis B/inmunología , Antígenos del Núcleo de la Hepatitis B/metabolismo , Antígenos de Superficie de la Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/metabolismo , Antígenos e de la Hepatitis B/sangre , Antígenos e de la Hepatitis B/inmunología , Virus de la Hepatitis B/efectos de los fármacos , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/metabolismo , Humanos , Inmunohistoquímica , Masculino , Ratones , Regiones Promotoras Genéticas , ARN Viral , Transcripción Genética , Replicación ViralRESUMEN
The role of hepatitis B virus (HBV) X protein (HBx) in the regulation of HBV replication remains controversial. In the present study, the role of HBx in regulating HBV replication was initially investigated in both HepG2 and Huh7 in vitro cell lines with a transient transfection system. Next, the regions of HBx responsible for transcriptional transactivation and promotion of HBV replication were mapped in an HBV replication mouse model by in vivo transfection of a series of HBx expression plasmids. In an in vitro setting, HBx deï¬ciency had little effect on HBV replication in Huh7 cells, but impaired HBV replication in HepG2 cells. In an in vivo setting, HBx had a strong enhancing effect on HBV transcription and replication. For the C-terminal two-thirds of the protein (amino acids [aa] 51 to 154) was required for this function of HBx, and the regions spanning aa 52 to 72 and 88 to 154 were found to be important for the stimulatory function of HBx on HBV replication. In conclusion, the role of HBx in HBV replication regulation is affected by host cell type, and HBx has an important role in stimulating HBV transcription and replication in hepatocytes in vivo. Further, the transcriptional transactivation function of HBx may be crucial for its stimulatory effect on HBV transcription and replication.
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Regulación Viral de la Expresión Génica , Virus de la Hepatitis B/fisiología , Transactivadores/metabolismo , Transcripción Genética , Replicación Viral , Animales , Línea Celular , Análisis Mutacional de ADN , Virus de la Hepatitis B/genética , Hepatocitos/virología , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Transactivadores/genética , Proteínas Reguladoras y Accesorias ViralesRESUMEN
Hepatitis B virus (HBV) infection disrupt the innate immunity response, which may play an important role in the chronic mechanism, while retinoic acid-induced gene I (RIG-I) mediated signaling pathway is one of the most important channel in the innate immunity. HBx and HBV polymerase may disrupt RIG-I mediated signaling pathway. The recent advances about HBV and RIG-I are reviewed in this article.
Asunto(s)
ARN Helicasas DEAD-box/metabolismo , Hepatitis B/inmunología , Inmunidad Innata/inmunología , Transducción de Señal , Proteína 58 DEAD Box , Productos del Gen pol/metabolismo , Humanos , Receptores Inmunológicos , Transactivadores/metabolismo , Proteínas Reguladoras y Accesorias ViralesRESUMEN
BACKGROUND: The effects of Hepatitis B virus (HBV) rtA181T/sW172* mutation on viral replication and pathogenicity was concerned recently. This study aimed to investigate the biological characteristics of rtA181T/sW172* mutant strain of HBV in animal model. METHODS: The rtA181T/sW172* mutant plasmid was constructed using the pHBV4.1 (wild type HBV) as a template. The wild and mutant HBV replication mouse models were established utilizing a hydrodynamic technique. The titers of hepatitis B surface antigen (HBsAg), hepatitis B e antigen, and HBV DNA in serum, and the levels of HBsAg, hepatitis B core antigen(HBcAg), HBV DNA replication intermediates (HBV DNA RI) and HBV RNA in liver were measured after 1, 3, 5, 7, 10, 12 and 15 days of plasmid injection. RESULTS: In wild-type HBV replication mouse model, serum HBsAg was high on day 1, 3, and 5, but became lower since day 7; while in mutant HBV mouse model, serum HBsAg was always at very low level. In liver tissues, HBV DNA RI of wild type HBV was detected on day 1 after transfection. The level subsequently peaked on day 3, gradually declined after day 5, and was almost undetectable on day 10. However, the HBV DNA RI levels of the mutant strain were always higher and lasted longer until day 15. Consistently, the expression levels of HBsAg and HBcAg in liver of the mutant group were significantly increased. CONCLUSIONS: In the case of the HBV rtA181T/sW172* mutation, the secretion of serum HBsAg was impaired, whereas HBV DNA replication and HBsAg/HBcAg expression were increased in liver. These results suggest that the mutation can impair HBsAg secretion, and may cause the accumulation of viral core particles in liver.