RESUMEN
The emergence of SARS-CoV-2 variants with mutations in major neutralizing antibody-binding sites can affect humoral immunity induced by infection or vaccination1-6. Here we analysed the development of anti-SARS-CoV-2 antibody and T cell responses in individuals who were previously infected (recovered) or uninfected (naive) and received mRNA vaccines to SARS-CoV-2. While individuals who were previously infected sustained higher antibody titres than individuals who were uninfected post-vaccination, the latter reached comparable levels of neutralization responses to the ancestral strain after the second vaccine dose. T cell activation markers measured upon spike or nucleocapsid peptide in vitro stimulation showed a progressive increase after vaccination. Comprehensive analysis of plasma neutralization using 16 authentic isolates of distinct locally circulating SARS-CoV-2 variants revealed a range of reduction in the neutralization capacity associated with specific mutations in the spike gene: lineages with E484K and N501Y/T (for example, B.1.351 and P.1) had the greatest reduction, followed by lineages with L452R (for example, B.1.617.2). While both groups retained neutralization capacity against all variants, plasma from individuals who were previously infected and vaccinated displayed overall better neutralization capacity than plasma from individuals who were uninfected and also received two vaccine doses, pointing to vaccine boosters as a relevant future strategy to alleviate the effect of emerging variants on antibody neutralizing activity.
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Anticuerpos Antivirales/inmunología , COVID-19/epidemiología , COVID-19/virología , SARS-CoV-2/inmunología , Linfocitos T/inmunología , Vacunas Sintéticas/inmunología , Vacunas de ARNm/inmunología , Vacuna nCoV-2019 mRNA-1273/inmunología , Adulto , Anciano , Anticuerpos Neutralizantes/inmunología , Vacuna BNT162/inmunología , Femenino , Personal de Salud/estadística & datos numéricos , Humanos , Inmunidad Humoral , Masculino , Persona de Mediana Edad , Mutación , Estudios Retrospectivos , SARS-CoV-2/clasificación , Glicoproteína de la Espiga del Coronavirus/genética , Glicoproteína de la Espiga del Coronavirus/inmunologíaRESUMEN
The impact of Coronavirus Disease 2019 (COVID-19) mRNA vaccination on pregnancy and fertility has become a major topic of public interest. We investigated 2 of the most widely propagated claims to determine (1) whether COVID-19 mRNA vaccination of mice during early pregnancy is associated with an increased incidence of birth defects or growth abnormalities; and (2) whether COVID-19 mRNA-vaccinated human volunteers exhibit elevated levels of antibodies to the human placental protein syncytin-1. Using a mouse model, we found that intramuscular COVID-19 mRNA vaccination during early pregnancy at gestational age E7.5 did not lead to differences in fetal size by crown-rump length or weight at term, nor did we observe any gross birth defects. In contrast, injection of the TLR3 agonist and double-stranded RNA mimic polyinosinic-polycytidylic acid, or poly(I:C), impacted growth in utero leading to reduced fetal size. No overt maternal illness following either vaccination or poly(I:C) exposure was observed. We also found that term fetuses from these murine pregnancies vaccinated prior to the formation of the definitive placenta exhibit high circulating levels of anti-spike and anti-receptor-binding domain (anti-RBD) antibodies to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) consistent with maternal antibody status, indicating transplacental transfer in the later stages of pregnancy after early immunization. Finally, we did not detect increased levels of circulating anti-syncytin-1 antibodies in a cohort of COVID-19 vaccinated adults compared to unvaccinated adults by ELISA. Our findings contradict popular claims associating COVID-19 mRNA vaccination with infertility and adverse neonatal outcomes.
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COVID-19 , Animales , Anticuerpos Antivirales , COVID-19/prevención & control , Femenino , Feto , Productos del Gen env , Humanos , Ratones , Placenta/metabolismo , Embarazo , Proteínas Gestacionales , ARN Mensajero/genética , ARN Mensajero/metabolismo , SARS-CoV-2 , VacunaciónRESUMEN
BACKGROUND: Miscarriage is a frustrating complication of pregnancy that is common among women of reproductive age. Insufficient decidualization which not only impairs embryo implantation but disturbs fetomaternal immune-tolerance, has been widely regarded as a major cause of miscarriage; however, the underlying mechanisms resulting in decidual impairment are largely unknown. METHODS: With informed consent, decidual tissue from patients with spontaneous abortion or normal pregnant women was collected to detect the expression profile of UCHL1. Human endometrial stromal cells (HESCs) were used to explore the roles of UCHL1 in decidualization and dNK modulation, as well as the mechanisms involved. C57/BL6 female mice (7-10 weeks old) were used to construct pregnancy model or artificially induced decidualization model to evaluate the effect of UCHL1 on mice decidualization and pregnancy outcome. RESULTS: The Ubiquitin C-terminal hydrolase L1 (UCHL1), as a deubiquitinating enzyme, was significantly downregulated in decidua from patients with miscarriage, along with impaired decidualization and decreased dNKs. Blockage of UCHL1 led to insufficient decidualization and resultant decreased expression of cytokines CXCL12, IL-15, TGF-ß which were critical for generation of decidual NK cells (dNKs), whereas UCHL1 overexpression enhanced decidualization accompanied by increase in dNKs. Mechanistically, the promotion of UCHL1 on decidualization was dependent on its deubiquitinating activity, and intervention of UCHL1 inhibited the activation of JAK2/STAT3 signaling pathway, resulting in aberrant decidualization and decreased production of cytokines associated with dNKs modulation. Furthermore, we found that inhibition of UCHL1 also disrupted the decidualization in mice and eventually caused adverse pregnancy outcome. CONCLUSIONS: UCHL1 plays significant roles in decidualization and dNKs modulation during pregnancy in both humans and mice. Its deficiency indicates a poor pregnancy outcome due to defective decidualization, making UCHL1 a potential target for the diagnosis and treatment of miscarriage.
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Aborto Espontáneo , Decidua , Células Asesinas Naturales , Ratones Endogámicos C57BL , Ubiquitina Tiolesterasa , Ubiquitina Tiolesterasa/metabolismo , Ubiquitina Tiolesterasa/deficiencia , Femenino , Decidua/metabolismo , Animales , Embarazo , Aborto Espontáneo/metabolismo , Humanos , Células Asesinas Naturales/metabolismo , Células Asesinas Naturales/inmunología , Adulto , Ratones , Células del Estroma/metabolismo , Transducción de SeñalRESUMEN
Taxus stem barks can be used for extraction of paclitaxel. However, the composition of taxoids across the whole stem and the stem tissue-specificity of paclitaxel biosynthesis-related enzymes remain largely unknown. We used cultivated Taxus media trees for analyses of the chemical composition and protein of major stem tissues by an integrated metabolomic and proteomic approach, and the role of TmMYB3 in paclitaxel biosynthesis was investigated. The metabolomic landscape analysis showed differences in stem tissue-specific accumulation of metabolites. Phytochemical analysis revealed that there is high accumulation of paclitaxel in the phloem. Ten key enzymes involved in paclitaxel biosynthesis were identified, most of which are predominantly produced in the phloem. The full-length sequence of TmMYB3 and partial promoter sequences of five paclitaxel biosynthesis-related genes were isolated. Several MYB recognition elements were found in the promoters of TBT, DBTNBT and TS. Further in vitro and in vivo investigations indicated that TmMYB3 is involved in paclitaxel biosynthesis by activating the expression of TBT and TS. Differences in the taxoid composition of different stem tissues suggest that the whole stem of T. media has potential for biotechnological applications. Phloem-specific TmMYB3 plays a role in the transcriptional regulation of paclitaxel biosynthesis, and may explain the phloem-specific accumulation of paclitaxel.
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Paclitaxel/biosíntesis , Floema/metabolismo , Proteínas de Plantas/metabolismo , Tallos de la Planta/metabolismo , Taxus/metabolismo , Factores de Transcripción/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Redes y Vías Metabólicas , Metabolómica , Proteínas de Plantas/fisiología , Regiones Promotoras Genéticas , Proteómica , Factores de Transcripción/fisiologíaRESUMEN
BACKGROUND: Taxol is an efficient anticancer drug accumulated in Taxus species. Pseudotaxus chienii is an important member of Taxaceae, however, the level of six taxoids in P. chienii is largely unknown. RESULTS: High accumulation of 10-DAB, taxol, and 7-E-PTX suggested that P. chienii is a good taxol-yielding species for large-scale cultivation. By the omics approaches, a total of 3,387 metabolites and 61,146 unigenes were detected and annotated. Compared with a representative Taxus tree (Taxus yunnanensis), most of the differentially accumulated metabolites and differential expressed genes were assigned into 10 primary and secondary metabolism pathways. Comparative analyses revealed the variations in the precursors and intermediate products of taxol biosynthesis between P. chienii and T. yunnanensis. Taxusin-like metabolites highly accumulated in P. chienii, suggesting a wider value of P. chienii in pharmaceutical industry. CONCLUSIONS: In our study, the occurrence of taxoids in P. chienii was determined. The differential expression of key genes involved in the taxol biosynthesis pathway is the major cause of the differential accumulation of taxoids. Moreover, identification of a number of differentially expressed transcription factors provided more candidate regulators of taxol biosynthesis. Our study may help to reveal the differences between Pseudotaxus and Taxus trees, and promote resource utilization of the endangered and rarely studied P. chienii.
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Vías Biosintéticas , Metaboloma , Metabolómica , Paclitaxel/biosíntesis , Plantas Medicinales/metabolismo , Especificidad de la Especie , Taxaceae/metabolismo , Especies en Peligro de Extinción , Variación GenéticaRESUMEN
Focal inflammation and remyelination failure are major hallmarks of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). In this study, we found that leonurine, a bioactive alkaloid, alleviated EAE disease severity along with reduced central nervous system inflammation and myelin damage. During the pathogenesis of EAE, leonurine dramatically suppressed the recruitment of encephalitogenic T cells into the central nervous system, whereas did not impair periphery immune responses and microglia activation. Mechanistically, leonurine protected mice against demyelination along with enhanced remyelination through promoting the maturation of oligodendrocytes in both EAE and cuprizone-induced demyelination mouse models. Moreover, we identified that the expression of demethylase jumonji domain-containing protein D3 was significantly enhanced upon treatment of leonurine, which suppressed the trimethylation of histone H3 lysine-27 and enhanced oligodendrocyte maturation accordingly. Collectively, our study identified the therapeutic effect of leonurine on EAE model, which potentially represents a promising therapeutic strategy for multiple sclerosis, even other demyelination disorders.
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Ácido Gálico/análogos & derivados , Inflamación/tratamiento farmacológico , Histona Demetilasas con Dominio de Jumonji/genética , Esclerosis Múltiple/tratamiento farmacológico , Animales , Diferenciación Celular/efectos de los fármacos , Sistema Nervioso Central/efectos de los fármacos , Sistema Nervioso Central/patología , Cuprizona/toxicidad , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental , Ácido Gálico/farmacología , Humanos , Inflamación/inducido químicamente , Inflamación/genética , Inflamación/patología , Ratones , Microglía/efectos de los fármacos , Microglía/patología , Esclerosis Múltiple/genética , Esclerosis Múltiple/patología , Vaina de Mielina/genética , Neurogénesis/efectos de los fármacos , Oligodendroglía/metabolismo , Oligodendroglía/patología , Remielinización/efectos de los fármacosRESUMEN
Modulating T-cell alloreactivity has been a main strategy to reduce graft-versus-host disease (GVHD), a life-threatening complication after allogeneic hematopoietic stem-cell transplantation (HSCT). Genetic deletion of T-cell Ezh2, which catalyzes trimethylation of histone H3 at lysine 27 (H3K27me3), inhibits GVHD. Therefore, reducing Ezh2-mediated H3K27me3 is thought to be essential for inhibiting GVHD. We tested this hypothesis in mouse GVHD models. Unexpectedly, administration of the Ezh2 inhibitor GSK126, which specifically decreases H3K27me3 without affecting Ezh2 protein, failed to prevent the disease. In contrast, destabilizing T-cell Ezh2 protein by inhibiting Hsp90 using its specific inhibitor AUY922 reduced GVHD in mice undergoing allogeneic HSCT. In vivo administration of AUY922 selectively induced apoptosis of activated T cells and decreased the production of effector cells producing interferon γ and tumor necrosis factor α, similar to genetic deletion of Ezh2. Introduction of Ezh2 into alloreactive T cells restored their expansion and production of effector cytokines upon AUY922 treatment, suggesting that impaired T-cell alloreactivity by inhibiting Hsp90 is achieved mainly through depleting Ezh2. Mechanistic analysis revealed that the enzymatic SET domain of Ezh2 directly interacted with Hsp90 to prevent Ezh2 from rapid degradation in activated T cells. Importantly, pharmacological inhibition of Hsp90 preserved antileukemia activity of donor T cells, leading to improved overall survival of recipient mice after allogeneic HSCT. Our findings identify the Ezh2-Hsp90 interaction as a previously unrecognized mechanism essential for T-cell responses and an effective target for controlling GVHD.
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Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Enfermedad Injerto contra Huésped/inmunología , Enfermedad Injerto contra Huésped/prevención & control , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Linfocitos T/inmunología , Animales , Proteína Potenciadora del Homólogo Zeste 2/química , Proteínas HSP90 de Choque Térmico/metabolismo , Hematopoyesis/efectos de los fármacos , Trasplante de Células Madre Hematopoyéticas , Histonas/metabolismo , Indoles/farmacología , Isoxazoles/farmacología , Lisina/metabolismo , Metilación/efectos de los fármacos , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Antígenos de Histocompatibilidad Menor/metabolismo , Dominios Proteicos , Estabilidad Proteica/efectos de los fármacos , Piridonas/farmacología , Resorcinoles/farmacología , Linfocitos T/efectos de los fármacos , Trasplante HomólogoRESUMEN
UNLABELLED: Fulminant hepatic failure (FHF) is a clinical syndrome characterized by sudden and severe impairment of liver function. Mesenchymal stem cells (MSCs) have been proposed as a promising therapeutic approach for FHF. In this study we used Propionibacterium acnes (P. acnes)-primed, lipopolysaccharide (LPS)-induced liver injury in mice as an animal model of human FHF. We demonstrated that administration of MSCs significantly ameliorated liver injury and improved the survival rates of mice subjected to P. acnes plus LPS-induced FHF. Allogeneic MSCs showed similar treatment efficacy as autologous MSCs did in FHF. Treatment efficacy of MSCs could be attributed to decreased infiltration and activation of CD4(+) T cells in the liver, inhibition of T helper 1 cells, and induction of regulatory T cells (Tregs). Moreover, decreased DNA copies of P. acnes were detected in the liver of MSC-treated mice. Intriguingly, a distinct liver population of CD11c(+) MHCII(hi) CD80(lo) CD86(lo) regulatory dendritic cells (DCs) was induced by MSCs. Moreover, these DCs induced Treg differentiation through transforming growth factor-ß production. Further mechanistic studies demonstrated that MSC-derived prostaglandin E2 and one of its receptors, EP4, played essential roles in the differentiation of CD11c(+) B220(-) DC precursors into regulatory DCs in a phosphoinositide 3-kinase-dependent manner. CONCLUSION: MSCs induce regulatory DCs from CD11c(+) B220(-) DC precursors. This study elucidates an immunoregulatory mechanism of MSCs and lays a foundation for application of MSCs in FHF therapy.
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Células Dendríticas/patología , Infecciones por Bacterias Grampositivas/complicaciones , Fallo Hepático Agudo/microbiología , Fallo Hepático Agudo/terapia , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/patología , Propionibacterium acnes , Animales , Tratamiento Basado en Trasplante de Células y Tejidos , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Lipopolisacáridos/efectos adversos , Hígado/metabolismo , Hígado/patología , Fallo Hepático Agudo/inducido químicamente , Ratones , Ratones Endogámicos C57BL , Subtipo EP4 de Receptores de Prostaglandina E/metabolismo , Transducción de Señal , Linfocitos T Reguladores/metabolismo , Linfocitos T Reguladores/patología , Células TH1/metabolismo , Células TH1/patología , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
China has become an emerging destination for international migration, especially in some Association of South East Asian Nations countries, but the situation of migrants seeking medical care in China remains unclear. A retrospective cross-sectional study was conducted in a hospital in Chongzuo, which provides medical services for foreigners, to investigate the situation of Vietnamese people seeking health care in Guangxi, China. Vietnamese patients who visited the hospital between 2018 and 2020 were included in the study. Demographic characteristics, clinical characteristics, characteristics of payment for medical costs, and characteristics of hospitalization were compared between outpatients and inpatients. In total, 778 Vietnamese outpatients and 173 inpatients were included in this study. The percentages of female outpatients and inpatients were 93.44% and 88.44% (χ2 = 5.133, Pâ =â .023), respectively. Approximately 30% of outpatients and 47% of inpatients visited the hospital due to obstetric needs. The proportions of outpatients with basic medical insurance for urban residents, basic medical insurance for urban employees, and new cooperative medical schemes were 28.02%, 3.21%, and 2.31%, respectively. In comparison, the proportion of inpatients with the above 3 types of medical insurance was 16.76%, 1.73%, and 2.31%, respectively. The proportion of different payments for medical costs between outpatients and inpatients were significantly different (χ2 = 24.404, Pâ <â .01). Middle-aged Vietnamese females in Guangxi, China, may have much greater healthcare needs. Their main medical demand is for obstetric services. Measurements should be taken to improve the health services targeting Vietnamese female, but the legitimacy of Vietnamese in Guangxi is a major prerequisite for them to access more and better healthcare services.
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Emigración e Inmigración , Necesidades y Demandas de Servicios de Salud , Seguro de Salud , Obstetricia , Pueblos del Sudeste Asiático , Femenino , Humanos , Persona de Mediana Edad , China/epidemiología , Estudios Transversales , Seguro de Salud/estadística & datos numéricos , Estudios Retrospectivos , Pueblos del Sudeste Asiático/etnología , Pueblos del Sudeste Asiático/estadística & datos numéricos , Vietnam/etnología , Necesidades y Demandas de Servicios de Salud/economía , Necesidades y Demandas de Servicios de Salud/estadística & datos numéricos , Migrantes/estadística & datos numéricos , Emigración e Inmigración/estadística & datos numéricos , Obstetricia/economía , Obstetricia/estadística & datos numéricos , Aceptación de la Atención de Salud , Accesibilidad a los Servicios de Salud/estadística & datos numéricosRESUMEN
INTRODUCTION: Mesenchymal stromal cell (MSC)-based cell therapy is a promising approach for various inflammatory disorders based on their immunosuppressive capacity. Osteopontin (OPN) regulates several cellular functions including tissue repair, bone metabolism and immune reaction. However, the biological function of OPN in regulating the immunosuppressive capacity of MSCs remains elusive. OBJECTIVES: This study aims to highlight the underlying mechanism of the proinflammatory cytokines affect the therapeutic ability of MSCs through OPN. METHODS: MSCs in response to the proinflammatory cytokines were collected to determine the expression profile of OPN. In vitro T-cell proliferation assays and gene editing were performed to check the role and mechanisms of OPN in regulating the immunosuppressive capacity of MSCs. Inflammatory disease mouse models were established to evaluate the effect of OPN on improving MSC-based immunotherapy. RESULTS: We observed that OPN, including its two isoforms iOPN and sOPN, was downregulated in MSCs upon proinflammatory cytokine stimulation. Interestingly, iOPN, but not sOPN, greatly enhanced the immunosuppressive activity of MSCs on T-cell proliferation and thus alleviated the inflammatory pathologies of hepatitis and colitis. Mechanistically, iOPN interacted with STAT1 and mediated its deubiquitination, thereby inducing the master immunosuppressive mediator inducible nitric oxide synthase (iNOS) in MSCs. In addition, iOPN expression was directly downregulated by activated STAT1, which formed a negative feedback loop to restrain MSC immunosuppressive capacity. CONCLUSION: Our findings demonstrated that iOPN expression modulation in MSCs is a novel strategy to improve MSC-based immunotherapy.
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Células Madre Mesenquimatosas , Osteopontina , Factor de Transcripción STAT1 , Animales , Femenino , Masculino , Ratones , Proliferación Celular , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/inmunología , Ratones Endogámicos C57BL , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Osteopontina/metabolismo , Osteopontina/genética , Factor de Transcripción STAT1/metabolismo , Linfocitos T/metabolismo , Linfocitos T/inmunologíaRESUMEN
The toxicity and stress caused by heavy metal contamination has become an important constraint to the growth and flourishing of trees. In particular, species belonging to the genus Taxus, which are the only natural source for the anti-tumor medicine paclitaxel, are known to be highly sensitive to environmental changes. To investigate the response of Taxus spp. to heavy metal stress, we analyzed the transcriptomic profiles of Taxus media trees exposed to cadmium (Cd2+). In total, six putative genes from the metal tolerance protein (MTP) family were identified in T. media, including two Cd2+ stress inducible TMP genes (TmMTP1, TmMTP11 and Taxus media). Secondary structure analyses predicted that TmMTP1 and TmMTP11, which are members of the Zn-CDF and Mn-CDF subfamily proteins, respectively, contained six and four classic transmembrane domains, respectively. The introduction of TmMTP1/11 into the ∆ycf1 yeast cadmium-sensitive mutant strain showed that TmMTP1/11 might regulate the accumulation of Cd2+ to yeast cells. To screen the upstream regulators, partial promoter sequences of the TmMTP1/11 genes were isolated using the chromosome walking method. Several myeloblastosis (MYB) recognition elements were identified in the promoters of these genes. Furthermore, two Cd2+-induced R2R3-MYB TFs, TmMYB16 and TmMYB123, were identified. Both in vitro and in vivo assays confirmed that TmMTB16/123 play a role in Cd2+ tolerance by activating and repressing the expression of TmMTP1/11 genes. The present study elucidated new regulatory mechanisms underlying the response to Cd stress and can contribute to the breeding of Taxus species with high environmental adaptability.
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Metales Pesados , Taxus , Cadmio/metabolismo , Taxus/genética , Taxus/metabolismo , Saccharomyces cerevisiae , Metales Pesados/metabolismo , Paclitaxel/metabolismoRESUMEN
Background: Simultaneous pancreas-kidney transplantation is an important treatment approach for diabetic renal insufficiency, but pancreatic arteriovenous thrombosis is among the early serious surgical complications that can lead to graft loss and even be fatal. Ultrasound is considered to be a safe and non-invasive approach, but it is often affected by intestinal gas interference and operator proficiency, partial thromboses may be easily missed. Computed tomography angiography (CTA) and computed tomography venography (CTV) are highly accurate but radiative, requiring the use of contrast agents. Methods: A total of 194 patients with end-stage diabetic nephropathy who underwent simultaneous pancreas-kidney transplantation from September 2016 to May 2021 were selected, among which 32 patients with highly suspected arteriovenous thrombosis were enrolled as the research subjects. All patients were examined by color Doppler ultrasonography, CTA and CTV. CTA and CTV are the gold standard for diagnostic imaging. The diagnostic value of color Doppler ultrasound, CTA and CTV in the diagnosis of pancreatic arteriovenous thrombosis was compared. and Kappa coefficient was used for consistency test. Results: Among the 32 patients with high clinical suspicion of transplanted pancreatic arteriovenous thrombosis after simultaneous pancreas-kidney transplantation, 9 patients were diagnosed by CTA/CTV and 10 patients were diagnosed by color Doppler ultrasonography, of which 2 cases were false positive and 1 case false negative. After transplantation, the normal diameter of the donor splenic vein was 3.96±0.16 mm. The difference in the diameter of the donor splenic vein between those with and without donor splenic vein thrombosis was statistically significant (P<0.05). The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of color Doppler ultrasound in the diagnosis of arteriovenous thrombosis were 88.9%, 91.3%, 90.6%, 80%, and 95.5%, respectively. There was no significant difference between color Doppler ultrasound diagnosis of arteriovenous thrombosis and CTA and CTV results (McNemar test P=1). The diagnosis of arteriovenous thrombosis by color Doppler ultrasonography was consistent with that of CTA and CTV (Kappa coefficient =0.776). Conclusions: Color Doppler ultrasonography has the advantages of safety and radiation-free, and can be used as the first choice for diagnosis of pancreatic arteriovenous thrombosis after simultaneous pancreas-kidney transplantation.
RESUMEN
Taxus trees are major natural sources for the extraction of taxol, an anti-cancer agent used worldwide. Taxus media is a dioecious woody tree with high taxol yield. However, the sexually dimorphic accumulation of taxoids in T. media is largely unknown. Our study revealed high accumulation of taxoids in female T. media trees using a UPLC-MS/MS method. Thereafter, many differential metabolites and genes between female and male T. media trees were identified using metabolomic and transcriptomic analyses, respectively. Most of the taxol-related genes were predominantly expressed in female trees. A female-specific R2R3-MYB transcription factor gene, TmMYB39, was identified. Furthermore, bimolecular fluorescence complementation and yeast two-hybrid assays suggested the potential interaction between TmMYB39 and TmbHLH13. Several taxol biosynthesis-related promoter sequences were isolated and used for the screening of MYB recognition elements. The electrophoretic mobility shift assay indicated that TmMYB39 could bind to the promoters of the GGPPS, T10OH, T13OH, and TBT genes. Interaction between TmMYB39 and TmbHLH13 transactivated the expression of the GGPPS and T10OH genes. TmMYB39 might function in the transcriptional regulation of taxol biosynthesis through an MYB-bHLH module. Our results give a potential explanation for the sexually dimorphic biosynthesis of taxol in T. media.
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Fulminant hepatic failure (FHF) is a potentially fatal liver disease that is associated with intrahepatic infiltration of inflammatory cells. As the receptor of polyunsaturated long chain fatty acids, GPR120 can regulate cell differentiation, proliferation, metabolism, and immune response. However, whether GPR120 is involved in FHF remains unknown. Using Propionibacterium acnes (P. acnes)-primed, LPS-induced FHF in mice, we found that interference with GPR120 activity using pharmacological agonist attenuated the severity of the liver injury and mortality of FHF in mice, while a lack of GPR120 exacerbated the disease. GPR120 activation potently alleviated FHF and led to decreased T helper (Th) 1 cell response and expansion of regulatory T cells (Tregs). Interestingly, GPR120 agonist didn't directly target T cells, but dramatically induced a distinct population of CD11c+MHC IIlowCD80lowCD86low regulatory DCs in the livers of FHF mice. GPR120 was found to restrict HIF-1α-dependent glycolysis. The augmented HIF-1α stabilization caused by GPR120 antagonism or deletion could be attenuated by the inhibition of ERK or by the activation of AMPK. Through the analysis of the clinical FHF, we further confirmed the activation of GPR120 was negatively associated with the severity in patients. Our findings indicated that GPR120 activation has therapeutic potential in FHF. Strategies to target GPR120 using agonists or free fatty acids (FFAs) may represent a novel approach to FHF treatment.
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Células Dendríticas/metabolismo , Fallo Hepático Agudo/genética , Receptores Acoplados a Proteínas G/metabolismo , Animales , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Femenino , Glucólisis , Humanos , RatonesRESUMEN
Macroautophagy has been implicated in modulating the therapeutic function of mesenchymal stromal cells (MSCs). However, the biological function of chaperone-mediated autophagy (CMA) in MSCs remains elusive. Here, we found that CMA was inhibited in MSCs in response to the proinflammatory cytokines interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). In addition, suppression of CMA by knocking down the CMA-related lysosomal receptor lysosomal-associated membrane protein 2 (LAMP-2A) in MSCs significantly enhanced the immunosuppressive effect of MSCs on T cell proliferation, and as expected, LAMP-2A overexpression in MSCs exerted the opposite effect on T cell proliferation. This effect of CMA on the immunosuppressive function of MSCs was attributed to its negative regulation of the expression of chemokine C-X-C motif ligand 10 (CXCL10), which recruits inflammatory cells, especially T cells, to MSCs, and inducible nitric oxide synthase (iNOS), which leads to the subsequent inhibition of T cell proliferation via nitric oxide (NO). Mechanistically, CMA inhibition dramatically promoted IFN-γ plus TNF-α-induced activation of NF-κB and STAT1, leading to the enhanced expression of CXCL10 and iNOS in MSCs. Furthermore, we found that IFN-γ plus TNF-α-induced AKT activation contributed to CMA inhibition in MSCs. More interestingly, CMA-deficient MSCs exhibited improved therapeutic efficacy in inflammatory liver injury. Taken together, our findings established CMA inhibition as a critical contributor to the immunosuppressive function of MSCs induced by inflammatory cytokines and highlighted a previously unknown function of CMA.
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Autofagia Mediada por Chaperones , Terapia de Inmunosupresión , Inflamación/inmunología , Inflamación/patología , Células Madre Mesenquimatosas/inmunología , Animales , Autofagia Mediada por Chaperones/efectos de los fármacos , Quimiocina CXCL10/metabolismo , Activación Enzimática/efectos de los fármacos , Interferón gamma/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factor de Transcripción STAT1/metabolismo , Bazo/metabolismo , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND & AIMS: Fulminant hepatitis (FH) is a clinical syndrome characterized by sudden and severe liver dysfunction. Dot1L, a histone methyltransferase, is implicated in various physiologic and pathologic processes, including transcription regulation and leukemia. However, the role of Dot1L in regulating inflammatory responses during FH remains elusive. METHODS: Propionibacterium acnes (P. acnes)-primed, lipopolysaccharides (LPS)-induced FH was established in C57BL/6 mice and was treated with the Dot1L inhibitor EPZ-5676. Myeloid derived suppressor cells (MDSCs) were depleted by anti-Gr-1 antibody to evaluate their therapeutic roles in Dot1L treatment of FH. Moreover, peripheral blood of patients suffered with FH and healthy controls was collected to determine the expression profile of Dot1L-SOCS1-iNOS axis in their MDSCs. RESULTS: Here we identified that EPZ-5676, pharmacological inhibitor of Dot1L, attenuated the liver injury of mice subjected to FH. Dot1L inhibition led to decreased T helper 1 cell response and expansion of regulatory T cells (Tregs) during FH. Interestingly, Dot1L inhibition didn't directly target T cells, but dramatically enhanced the immunosuppressive function of MDSCs. Mechanistically, Dot1L inhibition epigenetically suppressed SOCS1 expression, thus inducing inducible nitric oxide synthase (iNOS) expression in a STAT1-dependent manner. Moreover, in human samples, the levels of Dot1L and SOCS1 expression were upregulated in MDSCs, accompanied by decreased expression of iNOS in patients with FH, compared with healthy controls. CONCLUSIONS: Altogether, our findings established Dot1L as a critical regulator of MDSC immunosuppressive function for the first time, and highlighted the therapeutic potential of Dot1L inhibitor for FH treatment.
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Bencimidazoles/farmacología , N-Metiltransferasa de Histona-Lisina/antagonistas & inhibidores , Células Supresoras de Origen Mieloide/efectos de los fármacos , Animales , Femenino , N-Metiltransferasa de Histona-Lisina/metabolismo , Humanos , Ratones , Ratones Endogámicos C57BL , Células Supresoras de Origen Mieloide/metabolismo , Células Supresoras de Origen Mieloide/patologíaRESUMEN
Recent studies have provided insights into innate and adaptive immune dynamics in coronavirus disease 2019 (COVID-19). However, the exact features of antibody responses that govern COVID-19 disease outcomes remain unclear. In this study, we analyzed humoral immune responses in 229 patients with asymptomatic, mild, moderate and severe COVID-19 over time to probe the nature of antibody responses in disease severity and mortality. We observed a correlation between anti-spike (S) immunoglobulin G (IgG) levels, length of hospitalization and clinical parameters associated with worse clinical progression. Although high anti-S IgG levels correlated with worse disease severity, such correlation was time dependent. Deceased patients did not have higher overall humoral response than discharged patients. However, they mounted a robust, yet delayed, response, measured by anti-S, anti-receptor-binding domain IgG and neutralizing antibody (NAb) levels compared to survivors. Delayed seroconversion kinetics correlated with impaired viral control in deceased patients. Finally, although sera from 85% of patients displayed some neutralization capacity during their disease course, NAb generation before 14 d of disease onset emerged as a key factor for recovery. These data indicate that COVID-19 mortality does not correlate with the cross-sectional antiviral antibody levels per se but, rather, with the delayed kinetics of NAb production.
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Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , COVID-19/inmunología , Inmunoglobulina G/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Anciano , Anciano de 80 o más Años , COVID-19/mortalidad , COVID-19/prevención & control , Vacunas contra la COVID-19/uso terapéutico , Portador Sano/inmunología , Femenino , Humanos , Inmunidad Humoral , Cinética , Tiempo de Internación/estadística & datos numéricos , Masculino , Persona de Mediana Edad , SARS-CoV-2/inmunología , Índice de Severidad de la Enfermedad , Factores de TiempoRESUMEN
Rationale: Resistance to pemetrexed (PEM)-based chemotherapy is a major cause of progression in non-small cell lung cancer (NSCLC) patients. The deubiquitinating enzyme UCHL1 was recently found to play important roles in chemoresistance and tumor progression. However, the potential roles and mechanisms of UCHL1 in PEM resistance remain unclear. Methods: Bioinformatics analyses and immunohistochemistry were used to evaluate UCHL1 expression in NSCLC specimens. Kaplan-Meier analysis with the log-rank test was used for survival analyses. We established PEM-resistant NSCLC cell lines by exposing them to step-wise increases in PEM concentrations, and in vitro and in vivo assays were used to explore the roles and mechanisms of UCHL1 in PEM resistance using the NSCLC cells. Results: In chemoresistant tumors from NSCLC patients, UCHL1 was highly expressed and elevated UCHL1 expression was strongly associated with poor outcomes. Furthermore, UCHL1 expression was significantly upregulated in PEM-resistant NSCLC cells, while genetic silencing or inhibiting UCHL1 suppressed resistance to PEM and other drugs in NSCLC cells. Mechanistically, UCHL1 promoted PEM resistance in NSCLC by upregulating the expression of thymidylate synthase (TS), based on reduced TS expression after UCHL1 inhibition and re-emergence of PEM resistance upon TS restoration. Furthermore, UCHL1 upregulated TS expression, which mitigated PEM-induced DNA damage and cell cycle arrest in NSCLC cells, and also conferred resistance to PEM and other drugs. Conclusions: It appears that UCHL1 promotes PEM resistance by upregulating TS in NSCLC cells, which mitigated DNA damage and cell cycle arrest. Thus, UCHL1 may be a therapeutic target for overcoming PEM resistance in NSCLC patients.
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Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Enzimas Desubicuitinizantes/metabolismo , Neoplasias Pulmonares/metabolismo , Timidilato Sintasa/metabolismo , Ubiquitina Tiolesterasa/metabolismo , Células A549 , Animales , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Puntos de Control del Ciclo Celular/efectos de los fármacos , Puntos de Control del Ciclo Celular/fisiología , Línea Celular Tumoral , Daño del ADN/efectos de los fármacos , Daño del ADN/fisiología , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/fisiología , Células HEK293 , Humanos , Inmunohistoquímica/métodos , Neoplasias Pulmonares/tratamiento farmacológico , Masculino , Ratones Endogámicos BALB C , Persona de Mediana Edad , Pemetrexed/farmacología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiologíaRESUMEN
BACKGROUND: Intake of ω-3 PUFAs have been demonstrated to have positive effects on pregnancy outcome, whose receptor, GPR120, regulates several cellular functions including differentiation, metabolism and immune reaction. However, whether GPR120 is involved in decidualization and pregnancy remains unknown. METHODS: Decidua tissue from women with normal pregnancy and spontaneous abortion were collected to determine the expression profile of GPR120. Abortion mouse models and artificially induced deciduoma in mice were established to evaluate the effect of GPR120 on pregnancy outcome and in vivo decidualization. HESCs and primary DSCs were used to explore the roles of GPR120 in decidualization and mechanisms involved. FINDINGS: We found that GPR120 functioned to promote decidualization by upregulating glucose uptake and pentose-phosphate pathway (PPP) of human endometrial stromal cells. Firstly, the expression of GPR120 in decidua of spontaneous abortion was downregulated compared to normal decidua. Lack of GPR120 predisposed mice to LPS or RU486 induced abortion. Decidualization was augmented by GPR120 via improving GLUT1-mediated glucose uptake and G6PD- mediated PPP. FOXO1 was upregulated by GPR120 via activation of ERK1/2 and AMPK signaling and increased the expression of GLUT1. Furthermore, the expression of chemokines and cytokines in decidual stromal cells was enhanced by GPR120. Lastly, GPR120 agonist ameliorated LPS-induced abortion in the mice. INTERPRETATION: GPR120 plays significant roles in decidualization and the maintenance of pregnancy, which might be a potential target for diagnosis and treatment of spontaneous abortion. FUND: Ministry of Science and Technology of China, National Natural Science Foundation of China, the Program of Science and Technology Commission of Shanghai Municipality.
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Aborto Espontáneo/metabolismo , Decidua/metabolismo , Glucosa/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Aborto Espontáneo/inducido químicamente , Aborto Espontáneo/genética , Adulto , Animales , Línea Celular , Modelos Animales de Enfermedad , Regulación hacia Abajo , Ácidos Grasos Omega-3 , Femenino , Humanos , Lipopolisacáridos/efectos adversos , Ratones , Mifepristona/efectos adversos , Vía de Pentosa Fosfato , Embarazo , Células del Estroma/citología , Células del Estroma/metabolismoRESUMEN
Mesenchymal stem cells (MSCs), which are pluripotent cells with immunomodulatory properties, have been considered good candidates for the therapy of several immune disorders, such as inflammatory bowel diseases, concanavalin A-induced liver injury, and graft-versus-host disease. The embryo is a natural allograft to the maternal immune system. A successful pregnancy depends on the timely extinction of the inflammatory response induced by embryo implantation, followed by the switch to a tolerant immune microenvironment in both the uterus and the system. Excessive infiltration of immune cells and serious inflammatory responses are triggers for embryo rejection, which results in miscarriage. Here, we demonstrated that adoptive transfer of MSCs could prevent fetal loss in a lipopolysaccharide (LPS)-induced abortion model and immune response-mediated spontaneous abortion model. The immunosuppressive MSCs alleviated excessive inflammation by inhibiting CD4 + T cell proliferation and promoting the decidual macrophage switch to M2 in a tumor necrosis factor-stimulated gene-6 (TSG-6)-dependent manner. Cell-to-cell contact with proinflammatory macrophages increased the TSG-6 production by the MSCs, thereby enhancing the suppressive regulation of T cells and macrophages. Moreover, proinflammatory macrophages in contact with the MSCs upregulated the expression of CD200 on the stem cells and facilitated the reprogramming of macrophages towards an anti-inflammatory skew through the interaction of CD200 with CD200R on proinflammatory macrophages. Therefore, the results demonstrate that a TSG-6-mediated paracrine effect, reinforced by cell-to-cell contact between MSCs and proinflammatory macrophages, is involved in the mechanism of MSC-mediated abortion relief through the induction of immune tolerance. Our study also indicates the potential application of MSCs in clinical recurrent miscarriages.