RESUMEN
Increasing evidence highlights the role of bacteria in promoting tumorigenesis. The underlying mechanisms may be diverse and remain poorly understood. Here, we report that Salmonella infection leads to extensive de/acetylation changes in host cell proteins. The acetylation of mammalian cell division cycle 42 (CDC42), a member of the Rho family of GTPases involved in many crucial signaling pathways in cancer cells, is drastically reduced after bacterial infection. CDC42 is deacetylated by SIRT2 and acetylated by p300/CBP. Non-acetylated CDC42 at lysine 153 shows an impaired binding of its downstream effector PAK4 and an attenuated phosphorylation of p38 and JNK, consequently reduces cell apoptosis. The reduction in K153 acetylation also enhances the migration and invasion ability of colon cancer cells. The low level of K153 acetylation in patients with colorectal cancer (CRC) predicts a poor prognosis. Taken together, our findings suggest a new mechanism of bacterial infection-induced promotion of colorectal tumorigenesis by modulation of the CDC42-PAK axis through manipulation of CDC42 acetylation.
Asunto(s)
Neoplasias Colorrectales , Infecciones por Salmonella , Proteína de Unión al GTP cdc42 , Humanos , Acetilación , Carcinogénesis , Proteína de Unión al GTP cdc42/metabolismo , Transformación Celular Neoplásica , Quinasas p21 Activadas/metabolismo , Transducción de SeñalRESUMEN
At the end of 2019 Wuhan witnessed an outbreak of "atypical pneumonia" that later developed into a global pandemic. Metagenomic sequencing rapidly revealed the causative agent of this outbreak to be a novel coronavirus denoted SARS-CoV-2. To provide a snapshot of the pathogens in pneumonia-associated respiratory samples from Wuhan prior to the emergence of SARS-CoV-2, we collected bronchoalveolar lavage fluid samples from 408 patients presenting with pneumonia and acute respiratory infections at the Central Hospital of Wuhan between 2016 and 2017. Unbiased total RNA sequencing was performed to reveal their "total infectome", including viruses, bacteria and fungi. We identified 35 pathogen species, comprising 13 RNA viruses, 3 DNA viruses, 16 bacteria and 3 fungi, often at high abundance and including multiple co-infections (13.5%). SARS-CoV-2 was not present. These data depict a stable core infectome comprising common respiratory pathogens such as rhinoviruses and influenza viruses, an atypical respiratory virus (EV-D68), and a single case of a sporadic zoonotic pathogen-Chlamydia psittaci. Samples from patients experiencing respiratory disease on average had higher pathogen abundance than healthy controls. Phylogenetic analyses of individual pathogens revealed multiple origins and global transmission histories, highlighting the connectedness of the Wuhan population. This study provides a comprehensive overview of the pathogens associated with acute respiratory infections and pneumonia, which were more diverse and complex than obtained using targeted PCR or qPCR approaches. These data also suggest that SARS-CoV-2 or closely related viruses were absent from Wuhan in 2016-2017.
Asunto(s)
COVID-19/epidemiología , Brotes de Enfermedades , Neumonía/epidemiología , Infecciones del Sistema Respiratorio/epidemiología , SARS-CoV-2/aislamiento & purificación , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Líquido del Lavado Bronquioalveolar/microbiología , COVID-19/virología , China/epidemiología , Estudios de Cohortes , Femenino , Perfilación de la Expresión Génica , Humanos , Masculino , Metagenómica , Persona de Mediana Edad , Filogenia , Neumonía/microbiología , Infecciones del Sistema Respiratorio/microbiología , Adulto JovenRESUMEN
BACKGROUND AIMS: The considerable efficacy of B-cell maturation antigen-targeted chimeric antigen receptor (CAR)-T-cell therapy has been extensively demonstrated in the treatment of relapsed or refractory multiple myeloma. Nevertheless, in clinical practice, prolonged hematologic toxicity (PHT) extends hospital stay and impairs long-term survival. METHODS: This retrospective study reviewed 99 patients with relapsed or refractory multiple myeloma who underwent B-cell maturation antigen CAR-T-cell therapy at our institution between April 2018 and September 2021 (ChiCTR1800017404). RESULTS: Among 93 evaluable patients, the incidence of prolonged hematologic toxicities was high after CAR-T-cell infusion, including 38.71% (36/93) of patients with prolonged neutropenia, 22.58% (21/93) with prolonged anemia and 59.14% (55/93) with prolonged thrombocytopenia. In addition, 9.68% (9/93) of patients experienced prolonged pancytopenia. Our multivariate analyses identified that cytokine profiles were independent risk factors for PHTs, whereas a sufficient baseline hematopoietic function and high CD4/CD8 ratio of CAR-T cells were protective factors for PHTs after CAR-T-cell infusion. Subgroup analyses found that the kinetics of post-CAR-T hematologic parameters were primarily determined by the collective effects of cytokine release syndrome and baseline hematopoietic functions, and showed influential weights for the three lineages. CONCLUSIONS: Our findings improve the understanding of the impact of cytokines on hematopoietic functions, which could contribute to the mechanism investigation and exploration of potential intervention strategies.
Asunto(s)
Mieloma Múltiple , Receptores Quiméricos de Antígenos , Humanos , Mieloma Múltiple/terapia , Antígeno de Maduración de Linfocitos B , Estudios Retrospectivos , Inmunoterapia Adoptiva/efectos adversos , Citocinas , Tratamiento Basado en Trasplante de Células y TejidosRESUMEN
BACKGROUND: The exact pathophysiology of TS is still elusive. Previous studies have identified default mode networks (DMN) abnormalities in patients with TS. However, these literatures investigated the neural activity during the tic suppression, not a true resting-state. Therefore, this study aimed to reveal the neural mechanism of Tourette's syndrome (TS) from the perspective of topological organization and functional connectivity within the DMN by electroencephalography (EEG) in resting-state. METHODS: The study was conducted by analyzing the EEG data of TS patients with graph theory approaches. Thirty children with TS and thirty healthy controls (HCs) were recruited, and all subjects underwent resting-state EEG data acquisition. Functional connectivity within the DMN was calculated, and network properties were measured. RESULTS: A significantly lower connectivity in the neural activity of the TS patients in the ß band was found between the bilateral posterior cingulate cortex/retrosplenial cortex (t = -3.02, p < 0.05). Compared to HCs, the TS patients' local topological properties (degree centrality) in the left temporal lobe in the γ band were changed, while the global topological properties (global efficiency and local efficiency) in DMN exhibited no significant differences. It was also demonstrated that the degree centrality of the left temporal lobe in the γ band was positively related to the Yale Global Tic Severity Scale scores (r = 0.369, p = 0.045). CONCLUSIONS: The functional connectivity and topological properties of the DMN of TS patients were disrupted, and abnormal DMN topological property alterations might affect the severity of tic in TS patients. The abnormal topological properties of the DMN in TS patients may be due to abnormal functional connectivity alterations. The findings provide novel insight into the neural mechanism of TS patients.
Asunto(s)
Tics , Síndrome de Tourette , Niño , Humanos , Síndrome de Tourette/diagnóstico por imagen , Red en Modo Predeterminado , Giro del Cíngulo , Electroencefalografía , Imagen por Resonancia Magnética , Mapeo Encefálico , Encéfalo/diagnóstico por imagenRESUMEN
BACKGROUND: Influenza is a common illness for its high rates of morbidity and transmission. The implementation of non-pharmaceutical interventions (NPIs) during the COVID-19 pandemic to manage its dissemination could affect the transmission of influenza. METHODS: A retrospective analysis, between 2018 and 2023, was conducted to examine the incidence of influenza virus types A and B among patients in sentinel cities located in North or South China as well as in Wuhan City. For validations, data on the total count of influenza patients from 2018 to 2023 were collected at the Central Hospital of Wuhan, which is not included in the sentinel hospital network. Time series methods were utilized to examine seasonal patterns and to forecast future influenza trends. RESULTS: Northern and southern cities in China had earlier outbreaks during the NPIs period by about 8 weeks compared to the 2018-2019. The implementation of NPIs significantly reduced the influenza-like illness (ILI) rate and infection durations. Influenza B Victoria and H3N2 were the first circulating strains detected after the relaxation of NPIs, followed by H1N1 across mainland China. The SARIMA model predicted synchronized H1N1 outbreak cycles in North and South China, with H3N2 expected to occur in the summer in southern cities and in the winter in northern cities over the next 3 years. The ILI burden is expected to rise in both North and South China over the next 3 years, with higher ILI% levels in southern cities throughout the year, especially in winter, and in northern cities mainly during winter. In Wuhan City and the Central Hospital of Wuhan, influenza levels are projected to peak in the winter of 2024, with 2 smaller peaks expected during the summer of 2023. CONCLUSIONS: In this study, we report the impact of NPIs on future influenza trends in mainland China. We recommend that local governments encourage vaccination during the transition period between summer and winter to mitigate economic losses and mortality associated with influenza.
Asunto(s)
COVID-19 , Subtipo H1N1 del Virus de la Influenza A , Gripe Humana , Humanos , Gripe Humana/epidemiología , Gripe Humana/prevención & control , COVID-19/epidemiología , COVID-19/prevención & control , Subtipo H3N2 del Virus de la Influenza A , Pandemias/prevención & control , Estudios Retrospectivos , China/epidemiologíaRESUMEN
BACKGROUND: Foreign body aspiration (FBA) is a serious condition with high morbidity and mortality rates. Although chest radiography is generally the first radiologic modality used in diagnosis, a substantial percentage of foreign bodies are radiolucent in adults with diagnosis challenging. METHODS: Retrospective review of adult patients with FBA diagnosed by flexible electronic bronchoscopy from 2012 to 2022 collecting demographics, history, hospital presentation, radiographic, and operative details. Risk factors associated with radiolucent foreign body inhalation in adults were explored using appropriate statistical methods. RESULTS: Between 1 January 2012 and 1 January 2022, 114 adult patients diagnosed with FBA were enrolled. The median age of participants was 65 years (IQR 52-74). Multidetector computed tomography (MDCT) examinations identified 28 cases (25%) showing direct visualization of the foreign body (defined as the radiopaque group) and 86 cases (75%) in the radiolucent group. Multivariable stepwise linear regression analysis showed increased odds of radiolucent foreign body inhalation in adults associated with pneumonic patches in MDCT (OR 6.99; 95% CI 1.80-27.22; P = 0.005) and plants/meat foreign bodies (OR 6.17; 95% CI 1.12-33.96; P = 0.04). A witnessed choking history (OR 0.02; 95% CI 0-0.14; P < 0.001) was a protective factor of radiolucent foreign body inhalation in adults. CONCLUSIONS: Unlike radiopaque FBA, in those presenting with a suspected radiolucent foreign body aspiration, the diagnosis is far more challenging. Risk factors such as lacking a choking history, non-resolving pneumonia (pneumonic patches) in MDCT findings, and plants/meat foreign bodies may help in the early diagnosis of radiolucent foreign body inhalation in adults. Further prospective multicenter studies should be conducted to validate the findings.
Asunto(s)
Obstrucción de las Vías Aéreas , Cuerpos Extraños , Adulto , Anciano , Obstrucción de las Vías Aéreas/etiología , Broncoscopía/métodos , Cuerpos Extraños/complicaciones , Cuerpos Extraños/diagnóstico por imagen , Cuerpos Extraños/epidemiología , Humanos , Persona de Mediana Edad , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Marine polysaccharides originated from seaweeds, including agar, alginate, carrageenan, and fucoidan, possess various kinds of physiological activities and have been widely used in food, agricultural and medical areas. However, the application has been greatly limited by their poor solubility and low bioavailability. Thus marine oligosaccharides, as the degradation products of those polysaccharides, have drawn increasing attentions due to their obvious biological activities, good solubility and excellent bioavailability. This review will summarize the recent advances on the source, molecular structure and physiological activity of marine oligosaccharides, emphasizing their application as functional food additives. Furthermore, the relationship between the structure and the physiological activity of marine oligosaccharides is also elucidated and highlighted. The review concludes with an outlook toward potential applications for preparing the functional oligosaccharides in food biotechnology and agriculture fields.
Asunto(s)
Algas Marinas , Alginatos , Biotecnología , Oligosacáridos , PolisacáridosRESUMEN
The optimized diagnosis algorithm of Clostridioides difficile infection (CDI) is worldwide concerns. The purpose of this study was to assess the toxigenic C. difficile test performance and propose an optimal laboratory workflow for the diagnosis of CDI in mild virulent epidemic areas. Diarrhea samples collected from patients were analyzed by glutamate dehydrogenase (GDH), toxin AB (CDAB), and nucleic acid amplification test (NAAT). We assessed the performance of GDH, the GDH-CDAB algorithm, and the GDH-NAAT algorithm using toxigenic culture (TC) as a reference method. In this study, 186 diarrhea samples were collected. The numbers of TC-positive and TC-negative samples were 39 and 147, respectively. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and kappa of the GDH assay were 100%, 80.3%, 57.4%, 100%, and 0.63; of the GDH-CDAB algorithm were 48.7%, 97.3%, 82.6%, 87.7%, and 0.54; and of the GDH-NAAT algorithm were 74.4%, 100%, 100%, 93.6%, and 0.82, respectively. The GDH-NAAT algorithm has great concordance with TC in detecting toxigenic C. difficile (kappa = 0.82), while the sensitivity of the GDH-CDAB algorithm was too low to meet the demand of CDI diagnosis clinically. GDH-NAAT algorithm is recommended for the detection of toxigenic C. difficile with high specificity, increased sensitivity, and cost-effective.
Asunto(s)
Clostridioides difficile , Infecciones por Clostridium/diagnóstico , ADN Bacteriano/análisis , Diarrea/microbiología , Heces , Glutamato Deshidrogenasa/análisis , Adulto , Algoritmos , Proteínas Bacterianas/análisis , Toxinas Bacterianas/análisis , China/epidemiología , Clostridioides difficile/aislamiento & purificación , Clostridioides difficile/metabolismo , Infecciones por Clostridium/epidemiología , Enterotoxinas/análisis , Heces/química , Heces/microbiología , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
In this study, a series of acidic or alkaline polypeptide chains were designed and grafted onto DEG-AM resin using Fmoc solid-phase synthesis to study the relationship between enzyme conformation and carrier surface charge. ß-d-glucosidase (ßGase) was then immobilized onto these modified carriers by adsorption. Each form of immobilized ßGase showed decreasing specific activity compared to that of the free. It could be attributed to both the changes in the enzyme conformation and the decrease in mass transfer efficiency. The optimum temperature of free ßGase, DEG@B3-ßGase is 55 °C, which of DEG@A3-ßGase is 65 °C and they all have the highest activity at pH 5. The Ea values ââof free ßGase, DEG@A3-ßGase, and DEG@B3-ßGase are 0.546 kJ/mol, 0.224 kJ/mol, and 0.446 kJ/mol, and the Km values were 1.30 mmol/L, 1.44 mmol/L and 2.63 mmol/L, respectively. It shows that free ßGase and DEG@A3-ßGase are more similar. Meanwhile, the free ßGase (1.0 g/L, pH 5.0) stored at 4 °C has a shorter half-life (t1/2), which is only 9 days. However, the half-life of DEG@B3-ßGase and DEG@A3-ßGase is 20 days and over 60 days, indicating that the negative charged surface was conducive to maintenance of the structure and catalytic property of ßGase.
Asunto(s)
Enzimas Inmovilizadas/química , Temperatura , beta-Glucosidasa/química , Catálisis , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Propiedades de SuperficieRESUMEN
Ulvan, a kind of polyanionic heteropolysaccharide consisting of 3-sulfated rhamnose, uronic acids (iduronic acid and glucuronic acid) and xylose, has been widely applied in food and cosmetic industries. In addition, ulvan can be converted into fermentable monosaccharides through the cascade system of carbohydrate-active enzymes. Ulvan lyases can degrade ulvan into ulvan oligosaccharides, which is the first step in the fully degradation of ulvan. Various ulvan lyases have been cloned and characterized from marine bacteria and grouped into five polysaccharide lyase (PL) families, namely: PL24, PL25, PL28, PL37 and PL40 families. The elucidation of the biochemical characterization, action pattern and catalytic mechanism of ulvan lyase would definitely enhance our understanding of the deep utilization of marine bioresource and marine carbon cycling. In this review, we summarized the recent progresses about the source and biochemical characteristics of ulvan lyase. Additionally, the structural characteristics and catalytic mechanisms have been introduced in detail. This comprehensive information should be helpful regarding the application of ulvan lyases.
Asunto(s)
Polisacárido Liasas/química , Polisacárido Liasas/metabolismo , Catálisis , Ácido Glucurónico , Modelos Moleculares , Oligosacáridos , Polisacáridos , Análisis de SecuenciaRESUMEN
In this work, we investigated the functions of structural modules within alginate lyase by truncating an endo-type alginate lyase into two successive catalytic modules. The effects of module deletion on biochemical characteristics and product distributions were further investigated. The N-terminal module (Aly7B-CDI) exhibited no activity toward alginate, polyM or polyG, but the C-terminal module (Aly7B-CDII) retained its activity. The full-length enzyme (Aly7B) and its truncated counterpart (Aly7B-CDII) had similar substrate specificities, but Aly7B-CDII had lower activity. Moreover, the activity of Aly7B was much higher than Aly7B-CDII at 30°C. Aly7B-CDII, however, possessed higher optimal pH and better pH stability than the full-length enzyme. The final degradation products for Aly7B were unsaturated di-, tri- and tetra-oligosaccharides, and those for Aly7B-CDII were unsaturated mono-, di-, tri-, tetra- and penta-oligosaccharides. Therefore, the potential impact of the noncatalytic module Aly7B-CDI on the catalytic module Aly7B-CDII was further elucidated by characterizing Aly7B and its truncations. These data contribute to the functional understanding of these differing modules.
Asunto(s)
Alginatos/metabolismo , Biocatálisis , Liasas/metabolismo , Alginatos/química , Secuencia de Aminoácidos , Concentración de Iones de Hidrógeno , Liasas/química , Liasas/genética , Alineación de SecuenciaRESUMEN
BACKGROUND: The emergence of carbapenem-resistant Klebsiella pneumoniae (CR-KP) has become a significant problem worldwide and also being a major threat to children and newborns. Here we report an outbreak of NDM-1-producing K. pneumoniae in a neonatal unit. RESULTS: Six CR-KP strains, isolated from neonates with symptoms of infection, were identified using a VITEK-2 compact system, and the clinical data were retrieved from the electronic case records. In vitro susceptibility testing with broth dilution method showed that all six K. pneumoniae isolates were resistant to carbapenems and susceptible to colistin, aminoglycosides, fluoroquinolones and tigecycline. Based on the polymerase chain reaction results, each isolate was found to be blaNDM-1 gene positive. Clonal relationships were analysed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and showed that two different PFGE patterns were formed, which belonged to sequence types ST234 and ST1412. Plasmids carrying blaNDM-1 were successfully transferred from four of the six isolates to an Escherichia coli recipient through conjugative assays. S1-PFGE and Southern blot hybridization showed that four NDM-1-producing K. pneumoniae were clonal and carried blaNDM-1 on the same plasmid. The outbreak was effectively controlled by reducing the potential infection sources. All the patients were successfully treated and recovered after receiving an increased dose of carbapenems. Although the source of this outbreak was not clear, comprehensive measures were carried out and the outbreak was effectively controlled. CONCLUSIONS: ST234 and ST1412 of NDM-1-producing Klebsiella pneumoniae are the resistant clone spread in the neonatal unit, comprehensive infection control measures and optimized carbapenem therapy played an important role in controlling this NDM-1-producing K. pneumoniae outbreak.
Asunto(s)
Proteínas Bacterianas/metabolismo , Infección Hospitalaria/microbiología , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/aislamiento & purificación , beta-Lactamasas/metabolismo , Antibacterianos/administración & dosificación , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Proteínas Bacterianas/genética , Infección Hospitalaria/tratamiento farmacológico , Infección Hospitalaria/transmisión , Brotes de Enfermedades , Femenino , Humanos , Lactante , Infecciones por Klebsiella/tratamiento farmacológico , Infecciones por Klebsiella/transmisión , Klebsiella pneumoniae/clasificación , Klebsiella pneumoniae/enzimología , Klebsiella pneumoniae/genética , Masculino , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , beta-Lactamasas/genéticaRESUMEN
Carrageenan, a kind of linear sulfated polysaccharides consisting of D-galactose with alternating α-1,3 and ß-1,4 linkages, has been widely applied in the food and cosmetic industries as thickening and gelling agents due to excellent properties, such as gel-forming ability and chemical stability. It can be degraded by carrageenases to produce a series of even-numbered carrageenan oligosaccharides, which exhibit various fascinating functions, such as anti-inflammation, anti-coagulation, anti-tumor, and anti-thrombosis effects. Numerous carrageenases have been isolated and identified from various sources. The enzymes are grouped into three categories, namely κ-carrageenase, ι-carrageenase, and λ-carrageenase based on their substrate specificities and primary sequences, respectively. Elucidating the paradigm of the enzyme at every aspect would definitely enhance our understanding of the marine carbon cycling and natural evolution of glycoside hydrolases (GHs). The structural features of these enzymes have been fully illustrated, which will improve our knowledge of its catalytic mechanisms. In this review, we have summarized the recent progresses of major sources, category, and the enzyme's biochemical characteristics. Additionally, structural characteristics and catalytic mechanisms have been introduced in detail. We conclude with a brief discussion of the potential of the carrageenases in possible future applications in preparing functional oligosaccharides with versatile activities. This comprehensive information should be helpful regarding the application of carrageenases.
Asunto(s)
Proteínas Bacterianas , Glicósido Hidrolasas , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Carragenina/química , Carragenina/metabolismo , Glicósido Hidrolasas/química , Glicósido Hidrolasas/genética , Glicósido Hidrolasas/metabolismo , Análisis de SecuenciaRESUMEN
Alginate lyases play an essential role in the production of oligosaccharides by degrading alginate polysaccharide. Although many alginate lyases from various microorganisms have been characterized, reports on alginate lyases with special characteristics and commercial potential are still rather rare. In this study, a new alginate lyase, FsAlgA, was cloned from the deep-sea marine bacterium Flammeovirga sp. NJ-04. The recombinant enzyme was purified on Ni-NTA sepharose and then characterized in detail. It exhibited the highest activity (3343.7 U/mg) at pH 7.0 and 50 °C. Notably, the FsAlgA retained more than 80% of its maximum activity after incubation at 50 °C for 30 min, suggesting that FsAlgA was a heat-stable alginate lyase. Additionally, FsAlgA possessed broad substrate specificity, showing high activities toward both poly ß-D-mannuronate (polyM) and poly α-L-guluronate (polyG). Furthermore, the K m values of FsAlgA toward sodium alginate (0.48 mM) and polyG (0.94 mM) were lower than that toward polyM (1.42 mM). The TLC and ESI-MS analyses indicated that FsAlgA endolytically degraded alginate polysaccharide and released oligosaccharides with degree of polymerization (DP) of 2-5. Therefore, it may be a potent tool to produce alginate oligosaccharides with low DPs.
Asunto(s)
Proteínas Bacterianas/metabolismo , Bacteroidetes/enzimología , Polisacárido Liasas/metabolismo , Desnaturalización Proteica , Alginatos/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Bacteroidetes/genética , Bacteroidetes/metabolismo , Estabilidad de Enzimas , Ambientes Extremos , Ácido Glucurónico/metabolismo , Ácidos Hexurónicos/metabolismo , Polisacárido Liasas/química , Polisacárido Liasas/genética , Polisacáridos Bacterianos/metabolismo , Agua de Mar/microbiología , Especificidad por SustratoRESUMEN
BACKGROUND: MicroRNAs (miR, miRNAs) play pivotal roles in numerous physiological and pathophysiological contexts. We investigated whether miR-362-5p act as an oncogene in chronic myeloid leukaemia (CML) and aimed to understand its potential underlying mechanisms. METHODS: We compared the miR-362-5p expression levels between CML and non-CML cell lines, and between fresh blood samples from CML patients and normal healthy controls using quantitative real-time PCR (qPCR). Cell counting kit-8 (CCK-8) and Annexin V-FITC/PI analyses were used to measure the effects of miR-362-5p on proliferation and apoptosis, and Transwell assays were used to evaluate migration and invasion. A xenograft model was used to examine in vivo tumourigenicity. The potential target of miR-362-5p was confirmed by a luciferase reporter assay, qPCR and western blotting. Involvement of the JNK1/2 and P38 pathways was investigated by western blotting. RESULTS: miR-362-5p was up-regulated in CML cell lines and fresh blood samples from CML patients, and was associated with Growth arrest and DNA damage-inducible (GADD)45α down-regulation. Inhibition of miR-362-5p simultaneously repressed tumour growth and up-regulated GADD45α expression in a xenograft model. Consistently, the knockdown of GADD45α expression partially neutralized the effects of miR-362-5p inhibition. Furthermore study suggested that GADD45α mediated downstream the effects of miR-362-5p, which might indirectly regulates the activation of the JNK1/2 and P38 signalling pathways. CONCLUSION: miR-362-5p acts as an oncomiR that down-regulates GADD45α, which consequently activates the JNK1/2 and P38 signalling. This finding provides novel insights into CML leukaemogenesis and may help identify new diagnostic and therapeutic targets.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , MicroARNs/fisiología , Proteínas Nucleares/metabolismo , Animales , Apoptosis/genética , Apoptosis/fisiología , Ciclo Celular/genética , Ciclo Celular/fisiología , Proteínas de Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Movimiento Celular/fisiología , Proliferación Celular/genética , Proliferación Celular/fisiología , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Regulación Neoplásica de la Expresión Génica/fisiología , Células HEK293 , Humanos , Células K562 , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/genética , Proteínas Nucleares/genética , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Natural killer (NK) cell-based immunotherapy holds promise for cancer treatment; however, its efficacy remains limited, necessitating the development of alternative strategies. Here, we report that venetoclax, an FDA-approved BCL-2 inhibitor, directly activates NK cells, enhancing their cytotoxicity against acute myeloid leukemia (AML) both in vitro and in vivo, likely independent of BCL-2 inhibition. Through comprehensive approaches, including bulk and single-cell RNA sequencing, avidity measurement, and functional assays, we demonstrate that venetoclax increases the avidity of NK cells to AML cells and promotes lytic granule polarization during immunological synapse (IS) formation. Notably, we identify a distinct CD161lowCD218b+ NK cell subpopulation that exhibits remarkable sensitivity to venetoclax treatment. Furthermore, venetoclax promotes mitochondrial respiration and ATP synthesis via the NF-κB pathway, thereby facilitating IS formation in NK cells. Collectively, our findings establish venetoclax as a multifaceted immunometabolic modulator of NK cell function and provide a promising strategy for augmenting NK cell-based cancer immunotherapy.
Asunto(s)
Compuestos Bicíclicos Heterocíclicos con Puentes , Inmunoterapia Adoptiva , Células Asesinas Naturales , Leucemia Mieloide Aguda , Sulfonamidas , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Compuestos Bicíclicos Heterocíclicos con Puentes/uso terapéutico , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/efectos de los fármacos , Humanos , Sulfonamidas/farmacología , Animales , Leucemia Mieloide Aguda/terapia , Leucemia Mieloide Aguda/inmunología , Leucemia Mieloide Aguda/patología , Ratones , Inmunoterapia Adoptiva/métodos , Línea Celular Tumoral , FN-kappa B/metabolismo , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Ratones Endogámicos NODRESUMEN
OBJECTIVES: To develop a deep learning-based system for precise, robust, and fully automated segmentation of the mandibular canal on cone beam computed tomography (CBCT) images. METHODS: The system was developed on 536 CBCT scans (training set: 376, validation set: 80, testing set: 80) from one center and validated on an external dataset of 89 CBCT scans from 3 centers. Each scan was annotated using a multi-stage annotation method and refined by oral and maxillofacial radiologists. We proposed a three-step strategy for the mandibular canal segmentation: extraction of the region of interest based on 2D U-Net, global segmentation of the mandibular canal, and segmentation refinement based on 3D U-Net. RESULTS: The system consistently achieved accurate mandibular canal segmentation in the internal set (Dice similarity coefficient [DSC], 0.952; intersection over union [IoU], 0.912; average symmetric surface distance [ASSD], 0.046 mm; 95% Hausdorff distance [HD95], 0.325 mm) and the external set (DSC, 0.960; IoU, 0.924; ASSD, 0.040 mm; HD95, 0.288 mm). CONCLUSIONS: These results demonstrated the potential clinical application of this AI system in facilitating clinical workflows related to mandibular canal localization. CLINICAL SIGNIFICANCE: Accurate delineation of the mandibular canal on CBCT images is critical for implant placement, mandibular third molar extraction, and orthognathic surgery. This AI system enables accurate segmentation across different models, which could contribute to more efficient and precise dental automation systems.
Asunto(s)
Tomografía Computarizada de Haz Cónico , Imagenología Tridimensional , Mandíbula , Tomografía Computarizada de Haz Cónico/métodos , Humanos , Mandíbula/diagnóstico por imagen , Mandíbula/anatomía & histología , Imagenología Tridimensional/métodos , Aprendizaje Profundo , Procesamiento de Imagen Asistido por Computador/métodosRESUMEN
SUPPRESSOR OF MAX2 LIKE 1 (SMAX1) is a member of the SUPPRESSOR of MAX2 1LIKE family of genes and is known as a target protein of KARRIKIN INSENSITIVE2 (KAI2)-MORE AXILLARY BRANCHES2 (MAX2), which mediates karrikin signaling in Arabidopsis. SMAX1 plays a significant role in seed germination, hypocotyl elongation, and root hair development in Arabidopsis. SMAX1 has not yet been identified and characterized in woody plants. This study identified and characterized SsSMAX1 in Sapium sebiferum and found that SsSMAX1 was highly expressed in the seed, hypocotyl, and root tips of S. sebiferum. SsSMAX1 was functionally characterized by ectopic expression in Arabidopsis. SsSMAX1 overexpression lines of Arabidopsis showed significantly delayed seed germination and produced seedlings with longer hypocotyl and roots than wild-type and Atsmax1 functional mutants. SsSMAX1 overexpression lines of Arabidopsis also had broader and longer leaves and petioles than wild-type and Atsmax1, suggesting that SsSMAX1 is functionally conserved. This study characterizes the SMAX1 gene in a woody and commercially valuable bioenergy plant, Sapium sebiferum. The results of this study are beneficial to future research on the molecular biology of woody plants.