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Thermally activated delayed fluorescence (TADF) emitters with a high horizontal orientation are highly essential for improving the external quantum efficiency (EQE) of organic light-emitting diodes; however, pivotal molecular design strategies to improve the horizontal orientation of solution processable TADF emitters are still scarce and challenging. Herein, a phenyl bridge is adopted to connect the double TADF units, and a dimerized TADF dendrimer, D4CzBNPh-SF, is successfully constructed. Compared to counterpart with single TADF unit, the proof-of-the-concept molecule not only exhibits an improved horizontal dipole ratio (78%) due to the π-delocalization-induced extended molecular conjugation, but also displays a faster reversed intersystem crossing rate constant (6.08×106 s-1) and a high photoluminescence quantum yield of 95% in neat film. Consequently, the non-doped solution-processed device with D4CzBNPh-SF as the emitter, achieves an ultra-high maximum EQE of 32.6%, which remains at 26.6% under a luminance of 1000 cd/m2. Furthermore, using D4CzBNPh-SF as a sensitizer, the TADF-sensitized fluorescence device exhibits a high maximum EQE of 30.7% at a luminance of 575 cd/m2 and a full width at half maximum of 36 nm.
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Fungal infections have emerged as a major concern among immunocompromised patients, causing approximately 2 million deaths each year worldwide. However, the regulatory mechanisms underlying antifungal immunity remain elusive and require further investigation. The E3 ligase Trim26 belongs to the tripartite motif (Trim) protein family, which is involved in various biological processes, including cell proliferation, antiviral innate immunity, and inflammatory responses. Herein, we report that Trim26 exerts protective antifungal immune functions after fungal infection. Trim26-deficient mice are more susceptible to fungemia than their wild-type counterparts. Mechanistically, Trim26 restricts inflammatory neutrophils infiltration and limits proinflammatory cytokine production, which can attenuate kidney fungal load and renal damage during Candida infection. Trim26-deficient neutrophils showed higher proinflammatory cytokine expression and impaired fungicidal activity. We further demonstrated that excessive neutrophils infiltration in the kidney was because of the increased production of chemokines CXCL1 and CXCL2, which are mainly synthesized in the macrophages or dendritic cells of Trim26-deficient mice after Candida albicans infections. Together, our study findings unraveled the vital role of Trim26 in regulating antifungal immunity through the regulation of inflammatory neutrophils infiltration and proinflammatory cytokine and chemokine expression during candidiasis.
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Candidiasis , Neutrófilos , Animales , Ratones , Antifúngicos , Candida albicans , Candidiasis/metabolismo , Candidiasis/microbiología , Citocinas/metabolismo , Infiltración Neutrófila , Proteínas de Motivos Tripartitos , Ubiquitina-Proteína LigasasRESUMEN
The G protein-coupled receptor ADGRE5 (CD97) binds to various metabolites that play crucial regulatory roles in metabolism. However, its function in the antiviral innate immune response remains to be determined. In this study, we report that CD97 inhibits virus-induced type-I interferon (IFN-I) release and enhances RNA virus replication in cells and mice. CD97 was identified as a new negative regulator of the innate immune receptor RIG-I, and RIG-1 degradation led to the suppression of the IFN-I signaling pathway. Furthermore, overexpression of CD97 promoted the ubiquitination of RIG-I, resulting in its degradation, but did not impact its mRNA expression. Mechanistically, CD97 upregulates RNF125 expression to induce RNF125-mediated RIG-I degradation via K48-linked ubiquitination at Lys181 after RNA virus infection. Most importantly, CD97-deficient mice are more resistant than wild-type mice to RNA virus infection. We also found that sanguinarine-mediated inhibition of CD97 effectively blocks VSV and SARS-CoV-2 replication. These findings elucidate a previously unknown mechanism through which CD97 negatively regulates RIG-I in the antiviral innate immune response and provide a molecular basis for the development of new therapeutic strategies and the design of targeted antiviral agents.
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Infecciones por Virus ARN , Virus ARN , Animales , Ratones , Antivirales/farmacología , Proteína 58 DEAD Box/metabolismo , Inmunidad Innata , Receptores Acoplados a Proteínas G/metabolismo , Infecciones por Virus ARN/genética , Virus ARN/genética , Ubiquitina-Proteína Ligasas/metabolismo , UbiquitinaciónRESUMEN
STING (stimulator of interferon genes) exerts protective cellular responses to viral infection via induction of interferon production and autophagy. Here, we report the role of STING in modulating the immune responses toward fungal infection. Upon Candida albicans stimulation, STING transited alongside the endoplasmic reticulum (ER) to the phagosomes. In phagosomes, STING directly bound with Src via the N-terminal 18 amino acids of STING, and this binding prevented Src from recruiting and phosphorylating Syk. Consistently, Syk-associated signaling and production of pro-inflammatory cytokines and chemokines were increased in mouse BMDCs (bone-marrow-derived dendritic cells) lacking STING with fungal treatment. STING deficiency improved anti-fungal immunity in systemic C. albicans infection. Importantly, administration of the N-terminal 18-aa (amino acid) peptide of STING improved host outcomes in disseminated fungal infection. Overall, our study identifies a previously unrecognized function of STING in negatively regulating anti-fungal immune responses and offers a potential therapeutic strategy for controlling C. albicans infection.
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Nucleótidos , Transducción de Señal , Animales , Ratones , Citocinas/metabolismo , Inmunidad Innata , Interferones/metabolismo , Nucleótidos/metabolismo , Fagosomas/metabolismo , Fagosomas/microbiologíaRESUMEN
Though the flourishment of materials with multiple resonance (MR) in blue to green regions, red-emissive MR emitters are still rare in literatures, which definitely should be resolved for further applications. Herein, we report a simple molecular design strategy for the construction of pure-red MR emitters by conjugate charge transfer, which could greatly enhance the π-conjugation degree and charge-transfer property of the target molecule while maintaining the basic feature of MR, leading to a significant redshift of more than 128â nm compared to the selected parent MR core. The proof-of-concept emitter PPZ-BN exhibited a pure-red emission with a dominant peak at 613â nm and a small full-width-at-half-maximum of 0.16â eV (48â nm). The optimized organic light-emitting diode showed a high external quantum efficiency of 26.9 %, a small efficiency roll-off, and an excellent operation stability (LT99) of more than 43â hours at an initial luminance of 10 000â cd m-2 .
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In the current study, we assessed the relationship between mutations and the blood cell counts and early progression of patients with diffuse large-B cell lymphoma (DLBCL). A total of 109 patients with newly diagnosed DLBCL were included in this study. UBE2A mutation was only found in patients with bone marrow involvement. The mutations of ZNF608, SF3B1, DTX1, and NCOR2 were related to blood cell counts. NCOR2 mutations were only detected in patients of the noncomplete response group (PR + SD + PD). In addition, the mutations of ATM, BTG2, TBL1XR1, and TP53 were linked to lower PFS/OS rate, while SGK1, SCOS1, and NFKBIE were related to higher PFS/OS rate. Importantly, we identified that Ann Arbor stage (III-IV), B symptoms, absolute lymphocyte count (ALC) abnormity, and MTOR mutation were the four independent influencing factors of the 12-month progression of DLBCL patients. Overall, this study revealed that mutations were associated with the early progression of DLBCL.
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Proteínas Inmediatas-Precoces , Linfoma de Células B Grandes Difuso , Humanos , Pronóstico , Supervivencia sin Enfermedad , Linfoma de Células B Grandes Difuso/diagnóstico , Linfoma de Células B Grandes Difuso/genética , Recuento de Linfocitos , Mutación , Estudios Retrospectivos , Enzimas Ubiquitina-Conjugadoras , Proteínas Supresoras de TumorRESUMEN
Molecules with donor-spacer-acceptor configuration have been developed rapidly given their peculiar properties. How to utilize intermolecular interactions and charge transfers for solution-processed organic light-emitting diodes (OLEDs) greatly relies on molecular design strategy. Herein, soluble luminophores with D-spacer-A motif were constructed via shortening the alkyl chain from nonane to propane, where the alkyl chain was utilized as a spatial linker between the donor and acceptor. The alkyl chain blocks the molecular conjugation and induces the existence of aggregation-induced intermolecular CT emission, as well as the improved solubility and morphology in a solid-state film. In addition, the length of the alkyl chain affects the glass transition temperature, carrier transport and balance properties. The mCP-3C-TRZ with nonane as the spacer shows better thermal stability and bipolar carrier transport ability, so the corresponding solution-processable phosphorescent organic light-emitting diodes exhibit superior external quantum efficiency of 9.8% when using mCP-3C-TRZ as a host material. This work offers a promising strategy to establish a bipolar host via utilizing intermolecular charge transfer process in an aggregated state.
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BACKGROUND: Increasing evidence has convincingly shown that abnormal pre-mRNA splicing is implicated in the development of most human malignancies. Serine/arginine-rich protein kinase 1 (SRPK1), a key splicing regulator, is reported to be overexpressed in leukemia and other cancer types, which suggests the therapeutic potential of targeting SRPK1. METHODS: SRPK1 expression was measured in 41 ENKTL patients by immunohistochemistry and mRNA expression was analyzed by qRTâPCR. We knocked down SRPK1 expression in the ENKTL cell line YT by siRNA transfection and inhibited SRPK1 using inhibitors (SPHINX31 and SRPIN340) in YT cells and peripheral blood lymphocytes (PBLs) isolated from ENKTL patients to investigate its role in cell proliferation and apoptosis. Then, RNA-seq analysis was performed to predict the potential signaling pathway by which SRPK1 inhibition induces cell death and further verified this prediction by Western blotting. RESULTS: In the present study, we initially evaluated the clinical significance of SRPK1 in extranodal natural killer/T-cell lymphoma (ENKTL), a very aggressive subtype of non-Hodgkin lymphoma. The expression of SRPK1 in ENKLT patients was examined by immunohistochemistry and qRTâPCR, which revealed SRPK1 overexpression in more than 60% of ENKTL specimens and its association with worse survival. Cellular experiments using the human ENKTL cell line YT and PBLs from ENKTL patients, demonstrated that inhibition of SRPK1 suppressed cell proliferation and induced apoptosis. Subsequently, we investigated the downstream targets of SRPK1 by RNA-seq analysis and found that SRPK1 inhibition induced ATF4/CHOP pathway activation and AKT1 inhibition. Furthermore, ENKTL patients presenting high SRPK1 expression showed resistance to cisplatin-based chemotherapy. The association of SRPK1 expression with cisplatin resistance was also confirmed in YT cells. SRPK1 overexpression via pLVX-SRPK1 plasmid transfection dramatically decreased the sensitivity of YT cells to cisplatin, while siRNA-mediated SRPK1 knockdown or SRPK1 inhibitor treatment significantly increased cisplatin cytotoxicity. CONCLUSION: In summary, these results support that SRPK1 might be a useful clinical prognostic indicator and therapeutic target for ENKTL, especially for patients who relapse after cisplatin-based chemotherapies.
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Linfoma Extranodal de Células NK-T , Humanos , Linfoma Extranodal de Células NK-T/tratamiento farmacológico , Linfoma Extranodal de Células NK-T/genética , Linfoma Extranodal de Células NK-T/patología , Proteínas Serina-Treonina Quinasas/genética , Cisplatino/farmacología , ARN Interferente Pequeño/genética , Empalme del ARNRESUMEN
Strategies to enhance the ratio of the molecular horizontal emitting dipole orientation (Θ⥠) for thermally activated delayed fluorescence (TADF) emitters have unlocked the full potential of efficiencies for the evaporated devices, which, however, remain elusive for the solution-processed ones. Here, a strategic molecular design for solution processable TADF emitters featuring high Θ⥠s is proposed by attaching flexible chains ended with bipolar 9,9'-spirobi[fluorene] subunits as anchoring groups onto TADF emitting core. It's unveiled that the anchoring groups not only enhance the horizontal orientation via enlarging molecular planarity, but also benefit the high photoluminescence in pristine films. The corresponding non-doped solution processable OLEDs substantiate an unprecedented maximum external quantum efficiency (EQEmax )>30 %. Meanwhile, combining these compounds as TADF sensitizers, and multiple resonance final emitter, solution-processed OLEDs achieve an EQEmax of 25.6 % with a narrow full width at half maximum of 29â nm.
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BACKGROUND/OBJECTIVE: Bone marrow biopsy, the gold standard for the diagnosis of multiple myeloma (MM), has main limitation of the invasiveness. Here, we explored the diagnostic and prognostic values of circulating miR-1246 in patients with MM. MATERIAL AND METHODS: Ninety MM patients and 30 healthy donors (control group) were recruited in this study. The expression of miR-1246 in the peripheral blood samples was detected using qPCR. The receiver operating characteristic (ROC) curve was used to assess the diagnostic value of miR-1246 in MM. The Kaplan-Meier survival analyze was performed to evaluate the prognostic value of miR-1246. RESULTS: The expression level of serum miR-1246 from newly diagnosed MM patients was significantly higher than that of the control group. Circulating miR-1246 level was decreased after treatment in remission patients, but remained high levels in relapsed patients (P < 0.05). ROC analysis demonstrated that miR-1246 showed a high diagnostic value in MM with an area under the curve (AUC) of 0.952, the sensitivity of 87%, and the specificity of 95% [95% confidence interval (CI) 0.902-1.007; P < 0.001]. Kaplan-Meier analysis showed that the progression-free survival (PFS) (14.0 months vs. 26.5 months, P = 0.045) and overall survival (OS) (20.5 months vs. 55.5 months, P = 0.014) were significantly shorter in patients with high miR-1246 expression as compared with those in patients with miR-1246 low expression. Multiple Cox regression model analysis showed that circulating miR-1246 was an independent prognostic factor for PFS (HR 2.786, 95% CI: 1.420-5.467, P = 0.003) and OS (HR 2.995, 95% CI: 1.166-7.689, P = 0.023) in MM patients. CONCLUSION: This study demonstrates that circulating miR-1246 level is elevated in MM patients, which shows high values in the diagnosis and prognosis prediction in patients with MM.
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MicroARNs , Mieloma Múltiple , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Humanos , Estimación de Kaplan-Meier , MicroARNs/sangre , MicroARNs/genética , Mieloma Múltiple/sangre , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/genética , PronósticoRESUMEN
Primary pancreatic lymphoma (PPL) is an extremely rare malignant tumor that accounts for less than 2% of extranodal malignant lymphomas and 0.5% of pancreatic tumors. The most common symptoms of PPL are abdominal pain (83%), abdominal mass (58%), and weight loss (50%). The final diagnosis of PPL is based on cell histology, and the most common histological subtype is diffuse large B-cell lymphoma (DLBCL). The number of reported cases of primary pancreatic mucosa-associated lymphoid tissue (MALT) lymphoma is limited. The conversion rate of MALT lymphoma varies between 3% and 12%. However, the transformation of primary pancreatic MALT lymphoma to DLBCL has not been reported. We present a case of a 58-year-old man whose main symptoms were nausea and vomiting. Magnetic resonance imaging of the abdomen showed space-occupying lesions in the neck and body of the pancreas. Histopathological examination after surgical resection was diagnosed as primary pancreatic MALT lymphoma. Local radiotherapy was recommended and rejected by the patient's family. The disease progressed in a short time. Left supraclavicular lymph node resection biopsy and pathological examination showed DLBCL. After the initiation of chemotherapy, glaucoma appeared in the right eye during treatment, and chemotherapy was suspended. The disease progressed rapidly and the patient eventually died.
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Linfoma de Células B de la Zona Marginal , Linfoma de Células B Grandes Difuso , Neoplasias Pancreáticas , Biopsia , Humanos , Linfoma de Células B Grandes Difuso/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Páncreas , Neoplasias Pancreáticas/diagnóstico por imagenRESUMEN
TBK1 is an essential kinase for the innate immune response against viral infection. However, the key molecular mechanisms regulating the TBK1 activation remain elusive. Here, we identify PRMT1, a type I protein arginine methyltransferase, as an essential regulator of TBK1 activation. PRMT1 directly interacts with TBK1 and catalyzes asymmetric methylation of R54, R134, and R228 on TBK1. This modification enhances TBK1 oligomerization after viral infection, which subsequently promotes TBK1 phosphorylation and downstream type I interferon production. More important, myeloid-specific Prmt1 knockout mice are more susceptible to infection with DNA and RNA viruses than Prmt1fl/fl mice. Our findings reveal insights into the molecular regulation of TBK1 activation and demonstrate the essential function of protein arginine methylation in innate antiviral immunity.
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Arginina/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteína-Arginina N-Metiltransferasas/metabolismo , Animales , Línea Celular , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Humanos , Inmunidad Innata , Interferón Tipo I/genética , Interferón Tipo I/metabolismo , Metilación , Ratones , Ratones Noqueados , Fosforilación , Agregado de Proteínas , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , Proteína-Arginina N-Metiltransferasas/deficiencia , Proteína-Arginina N-Metiltransferasas/genética , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Vesiculovirus/fisiologíaRESUMEN
CARD9 is an essential adaptor protein in antifungal innate immunity mediated by C-type lectin receptors. The activity of CARD9 is critically regulated by ubiquitination; however, the deubiquitinases involved in CARD9 regulation remain incompletely understood. In this study, we identified ovarian tumor deubiquitinase 1 (OTUD1) as an essential regulator of CARD9. OTUD1 directly interacted with CARD9 and cleaved polyubiquitin chains from CARD9, leading to the activation of the canonical NF-κB and MAPK pathway. OTUD1 deficiency impaired CARD9-mediated signaling and inhibited the proinflammatory cytokine production following fungal stimulation. Importantly, Otud1 -/- mice were more susceptible to fungal infection than wild-type mice in vivo. Collectively, our results identify OTUD1 as an essential regulatory component for the CARD9 signaling pathway and antifungal innate immunity through deubiquitinating CARD9.
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Proteínas Adaptadoras de Señalización CARD/metabolismo , Candida albicans/fisiología , Candidiasis/inmunología , Enzimas Desubicuitinizantes/metabolismo , Neutrófilos/inmunología , Proteasas Ubiquitina-Específicas/metabolismo , Animales , Células Cultivadas , Citotoxicidad Inmunológica , Enzimas Desubicuitinizantes/genética , Modelos Animales de Enfermedad , Humanos , Inmunidad Innata , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , FN-kappa B/metabolismo , Células RAW 264.7 , Transducción de Señal , Proteasas Ubiquitina-Específicas/genética , UbiquitinaciónRESUMEN
As a common injury almost all cells face, DNA damage in oocytes-especially double-strand breaks (DSBs), which occur naturally during the first meiosis phase (meiosis I) due to synaptic complex separation-affects the fertilization ability of oocytes, instead of causing cancer (as in somatic cells). The mechanism of oocytes to effectively repair DSB damage has not yet been clearly studied, especially considering medically induced DSBs superimposed on naturally occurring DSBs in meiosis I. It was found that maturation rates decreased or increased, respectively corresponding with overexpression or interference of p21 in bovine oocytes. At the same time, the maturation rate of bovine oocytes decreased with a gradual increase in Zeocin dose, and the p21 expression in those immature oocytes changed significantly with the gradual increase in Zeocin dose (same as increased DSB intensity). Same as p21, the variation trend of ATM expression was consistent with the gradual increase in Zeocin dose. Furthermore, the oocytes demonstrated tolerance to DSBs during meiosis I, while the maturation rates decreased when the damage exceeded a certain threshold; according to which, it may be that ATM regulates the p53-p21 pathway to affect the completion of meiosis. In addition, nonhomologous recombination and cumulus cells are potentially involved in the process by which oocytes respond to DSB damage.
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Proteínas de la Ataxia Telangiectasia Mutada/genética , Roturas del ADN de Doble Cadena/efectos de los fármacos , Meiosis/efectos de los fármacos , Oocitos/crecimiento & desarrollo , Animales , Bleomicina/farmacología , Bovinos , Células del Cúmulo/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Reparación del ADN/efectos de los fármacos , Femenino , Técnicas de Maduración In Vitro de los Oocitos , Meiosis/genética , Oocitos/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
Purely organic luminescent materials concurrently exhibiting thermally activated delayed fluorescence (TADF) and aggregation-induced emission (AIE) features are in great demand due to their high efficiency in aggregation-state toward efficient nondoped OLEDs. Herein, a class of TADF emitters adopting phenyl(pyridyl)methanone as electron-accepting segments and di(tert-butyl)carbazole and 9,9-dimethyl-9,10-dihydroacridine (or phenoxazine) as electron-donating groups are designed and synthesized. The existence of intramolecular hydrogen bonding is conducive to minish the energy difference between a singlet and a triplet (ΔEst), suppress nonradiative decay, and increase the luminescence efficiency. By using 3CPyM-DMAC as the emitter, the nondoped device via a solution process realize a high current efficiency (CE) and external quantum efficiency (EQE) of 35.4 cd A-1 and 11.4%, respectively, which is superior to that of CBM-DMAC with a CE and EQE of 14.3 cd A-1 and 6.7%. This work demonstrates a promising tactic to the establishment of TADF emitters with AIE features via introducing intramolecular hydrogen bonding.
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@# Objective: To analyze the mutation of target genes in extranodal natural killer/T-cell lymphoma (ENKTL) by using nextgeneration sequencing, and to explore its relationship with prognosis and clinical characteristics, as to provide evidence for the pathogenesis, clinical diagnosis and targeted therapy of ENKTL. Methods: According to previous literature reports, the genes whose mutations can affect the development of lymphoma were selected as the target genes for this study. 29 patients with ENKTL, who were newly diagnosed at the Fourth Hospital of Hebei Medical University from August 2010 to October 2018, were selected for this study. The mutation of 9 target genes in the specimen was detected by thenext-generationsequencingtechnology.Therelationshipsamongclinicalfeatures,diseaseprognosisandmutationofthetargetgeneswereanalyzedbySPSS21.0statisticalsoftware.Results: :Ninetargetgenes were were screened. AT-rich interactive-domain 1A(ARID1A) gene showed the highest mutation rate in ENKTL (10 cases, 34.48%) followedbylysinemethyltransferase2D(KMT2D)gene(31.03%)andtumorprotein P53 (TP53) gene (24.13%). Kaplan-Meier survival analysis showed that the overall survival of ENKTL patients with KMT2D gene wild type was significantly better than patients with KMT2D gene mutation (P=0.006). The KMT2D gene mutation was found to besignificantlyrelatedtoclinicalstage,CRP,albumin,lymphocyte count and Ki67 expression in ENKTL patients (all P<0.05). COX regression analysis showed that KMT2D gene mutation was an independent adverse prognostic factor (P<0.05). Conclusion: The KMT2D gene has a high mutant frequency in ENKTL and is associated with patients’prognosis, suggesting that KMT2D gene plays an important role in the initiation and development of ENKTL. It could be used as a clinical therapeutic target for ENKTL.
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Bovine parainfluenza virus type 3 (BPIV3) is one of the most important pathogens associated with bovine respiratory diseases in both young and adult cattle widespreadly around the world. The host factors which participate in the infection of BPIV3 are poorly understood. Here, we found the bovine protein Cholesterol 25-hydroxylase (CH25â¯H) plays an important role in the infection of BPIV3. CH25H is a multi-transmembrane and endoplasmic reticulum-related enzyme that catalyzes oxidation reaction of cholesterol to production of 25-hydroxycholesterol (25HC) and significantly inhibits the replication of several viruses. In this study, we found that CH25H is an interferon-stimulated gene (ISG), which taken part in the antiviral innate immunity. In addition, the overexpression of CH25H could inhibit the replication of BPIV3, and 25HC significantly inhibited BPIV3 infection by preventing the synthesis of both virus antigenomic RNA (cRNA) and genomic RNA (gRNA) in MDBK cells. Interestingly, CH25H-M, a mutant lacking hydroxylase activity, still had an antiviral effect against BPIV3. Taken together, our findings highlight the antiviral function of CH25H during BPIV3 infection, and suggest that CH25H inhibits viral infection through both enzyme activity-dependent and -independent ways.
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Interacciones Huésped-Patógeno/fisiología , Virus de la Parainfluenza 3 Bovina/fisiología , Infecciones por Respirovirus/enzimología , Infecciones por Respirovirus/virología , Esteroide Hidroxilasas/genética , Esteroide Hidroxilasas/metabolismo , Replicación Viral/fisiología , Animales , Bovinos , Línea Celular , Expresión Génica/genética , Células HEK293 , Células HeLa , Humanos , Mutación , Replicación Viral/genéticaRESUMEN
BACKGROUND: Pasteurella multocida (P. multocida) is a bacterium that causes bovine respiratory disease (BRD) and haemorrhagic septicaemia (HS) in cattle, buffaloes and bison. Rapid point-of-care diagnosis or regular testing of Pasteurellosis, therefore, could contribute greatly to early detection, and screening infected animal is important. Up to now, there are no published reports on the use of recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) for P. multocida detection. RESULTS: This study proposes a promising isothermal detection method for P. multocida with the potential to be developed as an on-site test for Pasteurellosis. The method includes an RPA combined with LFD. First, the analytical sensitivity and specificity of P. multocida RPA-LFD were tested. The RPA-LFD, performed at 39 °C, successfully detected P. multocida DNA in 30 min, with a detection limit of up to 120 copies per reaction. Then, the practicability of RPA-LFD was analysed using 62 nasal swabs and 33 fresh lungs samples from 17 different dairy farms. Compared to real-time quantitative PCR (qPCR), the RPA-LFD assay yielded a clinical specificity of 95.15%, positive predictive value (PPV) of 95.15% and 0.958 kappa coefficient. Compared with the culture method, it achieved 100% sensitivity, 67.20% specificity and a 0.572 kappa coefficient. CONCLUSIONS: These results combined with the simple conditions required for the performance of the RPA-LFD assay, have demonstrated the effectiveness and practicability of the method for development into a regular on-site protocol for the diagnosis of Pasteurellosis.
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Enfermedades de los Bovinos/diagnóstico , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Infecciones por Pasteurella/veterinaria , Pasteurella multocida/aislamiento & purificación , Animales , Bovinos , ADN Bacteriano/genética , Pulmón/microbiología , Mucosa Nasal/microbiología , Técnicas de Amplificación de Ácido Nucleico/métodos , Infecciones por Pasteurella/diagnóstico , Pasteurella multocida/genética , Sensibilidad y EspecificidadRESUMEN
Bovine viral diarrhea virus (BVDV) is a worldwide spreading pestivirus affecting cattle and other ruminants; however, there have been few reports on epidemiologic investigation of BVDV in eastern China. In this study, bulk tank milk from 36 herds of dairy cattle in eastern China was submitted to serological investigations, 77.8% of herds was BVDV antibody positive. Individual animal status in two herds was further investigated collecting blood samples, the positive ratio was 49.74% and 24.64%, and the average positive ratio of calves, heifers, and lactating cows was 15.94%, 40.16%, and 41.7%, respectively. Moreover, clinical survey was carried out among 8170 dairy cattle from 36 herds, for diarrhea syndrome, respiratory problems and reproductive failure, and pathogens of all clinical cattle were further investigated. The results showed that BVDV was one of the main pathogen, which infected animals combining with various other viruses. Then, nine BVDV strains were isolated; phylogenetic analysis showed that BVDV subtypes currently circulating in eastern China were BVDV 1a and BVDV 1c. In addition, out of 377 cows tested, the 1.86% detected positive to the BVDV antigen. This study provided the foundation of further study on vaccination and control strategies of BVDV in eastern China.