RESUMEN
Single-cell analysis is a valuable tool for dissecting cellular heterogeneity in complex systems1. However, a comprehensive single-cell atlas has not been achieved for humans. Here we use single-cell mRNA sequencing to determine the cell-type composition of all major human organs and construct a scheme for the human cell landscape (HCL). We have uncovered a single-cell hierarchy for many tissues that have not been well characterized. We established a 'single-cell HCL analysis' pipeline that helps to define human cell identity. Finally, we performed a single-cell comparative analysis of landscapes from human and mouse to identify conserved genetic networks. We found that stem and progenitor cells exhibit strong transcriptomic stochasticity, whereas differentiated cells are more distinct. Our results provide a useful resource for the study of human biology.
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Células/citología , Células/metabolismo , Análisis de la Célula Individual/métodos , Adulto , Animales , Pueblo Asiatico , Diferenciación Celular , Línea Celular , Separación Celular , China , Bases de Datos Factuales , Cuerpos Embrioides/citología , Cuerpos Embrioides/metabolismo , Etnicidad , Feto/citología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Humanos , Inmunidad , Células Madre Pluripotentes Inducidas/citología , Células Madre Pluripotentes Inducidas/metabolismo , Islotes Pancreáticos/citología , Islotes Pancreáticos/metabolismo , Ratones , Especificidad de Órganos , ARN Mensajero/análisis , ARN Mensajero/genética , Análisis de Secuencia de ARN , Análisis de la Célula Individual/instrumentación , Procesos EstocásticosRESUMEN
Transforming growth factor-ß (TGF-ß) and Hippo signaling are two critical pathways engaged in cancer progression by regulating both oncogenes and tumor suppressors, yet how the two pathways coordinately exert their functions in the development of hepatocellular carcinoma (HCC) remains elusive. In this study, we firstly conducted an integrated analysis of public liver cancer databases and our experimental TGF-ß target genes, identifying CYR61 as a pivotal candidate gene relating to HCC development. The expression of CYR61 is downregulated in clinical HCC tissues and cell lines than that in the normal counterparts. Evidence revealed that CYR61 is a direct target gene of TGF-ß in liver cancer cells. In addition, TGF-ß-stimulated Smad2/3 and the Hippo pathway downstream effectors YAP and TEAD4 can form a protein complex on the promoter of CYR61, thereby activating the promoter activity and stimulating CYR61 gene transcription in a collaborative manner. Functionally, depletion of CYR61 enhanced TGF-ß- or YAP-mediated growth and migration of liver cancer cells. Consistently, ectopic expression of CYR61 was capable of impeding TGF-ß- or YAP-induced malignant transformation of HCC cells in vitro and attenuating HCC xenograft growth in nude mice. Finally, transcriptomic analysis indicates that CYR61 can elicit an antitumor program in liver cancer cells. Together, these results add new evidence for the crosstalk between TGF-ß and Hippo signaling and unveil an important tumor suppressor function of CYR61 in liver cancer.
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Carcinoma Hepatocelular , Proteína 61 Rica en Cisteína , Regulación Neoplásica de la Expresión Génica , Neoplasias Hepáticas , Factor de Crecimiento Transformador beta , Proteínas Señalizadoras YAP , Animales , Humanos , Ratones , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/genética , Línea Celular Tumoral , Movimiento Celular , Proteína 61 Rica en Cisteína/metabolismo , Proteína 61 Rica en Cisteína/genética , Minería de Datos , Regulación Neoplásica de la Expresión Génica/genética , Vía de Señalización Hippo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/genética , Ratones Desnudos , Regiones Promotoras Genéticas , Transducción de Señal/genética , Proteína Smad2/metabolismo , Proteína Smad2/genética , Proteína smad3/metabolismo , Proteína smad3/genética , Factores de Transcripción de Dominio TEA/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta/genética , Regulación hacia Arriba , Proteínas Señalizadoras YAP/metabolismo , Proteínas Señalizadoras YAP/genéticaRESUMEN
The mortality rate linked with nephrotic syndrome (NS) is quite high. The renal tubular injury influences the response of NS patients to steroid treatment. KN motif and ankyrin repeat domains 2 (KANK2) regulates actin polymerization, which is required for renal tubular cells to maintain their function. In this study, we found that the levels of KANK2 in patients with NS were considerably lower than those in healthy controls, especially in NS patients with acute kidney injury (AKI). To get a deeper understanding of the KANK2 transcriptional control mechanism, the core promoter region of the KANK2 gene was identified. KANK2 was further found to be positively regulated by E2F Transcription Factor 1 (E2F1), Transcription Factor AP-2 Gamma (TFAP2C), and Nuclear Respiratory Factor 1 (NRF1), both at mRNA and protein levels. Knocking down E2F1, TFAP2C, or NRF1 deformed the cytoskeleton of renal tubular cells and reduced F-actin content. EMSA and ChIP assays confirmed that all three transcription factors could bind to the upstream promoter transcription site of KANK2 to transactivate KANK2 in renal tubular epithelial cells. Our study suggests that E2F1, TFAP2C, and NRF1 play essential roles in regulating the KANK2 transcription, therefore shedding fresh light on the development of putative therapeutic options for the treatment of NS patients.
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Síndrome Nefrótico , Factor Nuclear 1 de Respiración , Humanos , Factor Nuclear 1 de Respiración/metabolismo , Síndrome Nefrótico/genética , Factores de Transcripción/metabolismo , Regulación de la Expresión Génica , Regiones Promotoras Genéticas/genética , Factor de Transcripción E2F1/genética , Factor de Transcripción AP-2/genéticaRESUMEN
Both exogenous gaseous and liquid forms of formaldehyde (FA) can induce depressive-like behaviors in both animals and humans. Stress and neuronal excitation can elicit brain FA generation. However, whether endogenous FA participates in depression occurrence remains largely unknown. In this study, we report that midbrain FA derived from lipopolysaccharide (LPS) is a direct trigger of depression. Using an acute depressive model in mice, we found that one-week intraperitoneal injection (i.p.) of LPS activated semicarbazide-sensitive amine oxidase (SSAO) leading to FA production from the midbrain vascular endothelium. In both in vitro and in vivo experiments, FA stimulated the production of cytokines such as IL-1ß, IL-6, and TNF-α. Strikingly, one-week microinfusion of FA as well as LPS into the midbrain dorsal raphe nucleus (DRN, a 5-HT-nergic nucleus) induced depressive-like behaviors and concurrent neuroinflammation. Conversely, NaHSO3 (a FA scavenger), improved depressive symptoms associated with a reduction in the levels of midbrain FA and cytokines. Moreover, the chronic depressive model of mice injected with four-week i.p. LPS exhibited a marked elevation in the levels of midbrain LPS accompanied by a substantial increase in the levels of FA and cytokines. Notably, four-week i.p. injection of FA as well as LPS elicited cytokine storm in the midbrain and disrupted the blood-brain barrier (BBB) by activating microglia and reducing the expression of claudin 5 (CLDN5, a protein with tight junctions in the BBB). However, the administration of 30 nm nano-packed coenzyme-Q10 (Q10, an endogenous FA scavenger), phototherapy (PT) utilizing 630-nm red light to degrade FA, and the combination of PT and Q10, reduced FA accumulation and neuroinflammation in the midbrain. Moreover, the combined therapy exhibited superior therapeutic efficacy in attenuating depressive symptoms compared to individual treatments. Thus, LPS-derived FA directly initiates depression onset, thereby suggesting that scavenging FA represents a promising strategy for depression treatment.
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Depresión , Lipopolisacáridos , Humanos , Ratones , Animales , Lipopolisacáridos/farmacología , Depresión/tratamiento farmacológico , Enfermedades Neuroinflamatorias , Citocinas/metabolismo , Mesencéfalo/metabolismo , FormaldehídoRESUMEN
Increasing evidences showed that ovulatory dysfunction, possibly caused by luteinized unruptured follicular follicle syndrome (LUFS), is one of the reasons for endometriosis-related infertility. The present study was conducted to explore the potential effect of elevated progesterone in follicular fluid (FF) on ovulation in endometriosis. A prospective study including 50 ovarian endometriosis patients and 50 control patients with matched pairs design was conducted with alterations in FF and peritoneal fluid (PF) components identified by metabolomics analyses and differentially expressed genes in granulosa cells (GCs) identified by transcriptome analysis. Patients with endometriosis exhibited a significantly higher progesterone level in serum, FF, and PF. Granulosa cells from endometriosis patients revealed decreased expression of HPGD, COX-2, and suppressed NF-ĸB signaling. Similarly, progesterone treatment in vitro downregulated HPGD and COX2 expression and suppressed NF-ĸB signaling in granulosa tumor-like cell line KGN (Bena Culture Collection, China) and primarily cultured GCs, as manifested by decreased expressions of IL1R1, IRAK3, reduced pIĸBα/IĸBα ratio, and nucleus translocation of p65. On the contrary, TNF-α treatment increased expression of IL1R1, IRAK3, pIĸBα, p65, and HPGD in GCs. One potential p65 binding site was identified in the promoter region of HPGD by chromatin immunoprecipitation. In conclusion, we found that intrafollicular progesterone might downregulate HPGD and COX-2 in GCs via suppressing the NF-ĸB signaling pathway, shedding light on the mechanism underlying the endometriosis-related ovulatory dysfunction.
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Endometriosis , Infertilidad Femenina , Femenino , Humanos , Progesterona/farmacología , Progesterona/metabolismo , Líquido Folicular/metabolismo , Endometriosis/genética , Endometriosis/metabolismo , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Estudios Prospectivos , Células de la Granulosa/metabolismo , Infertilidad Femenina/metabolismoRESUMEN
BACKGROUND: Colorectal cancer (CRC) is a leading cause of cancer-related mortality, highlighting an unmet clinical need for more effective therapies. This study aims to evaluate the causal relationship between 4,489 plasma proteins and CRC to identify potential therapeutic targets for CRC. METHODS: We conducted two-sample Mendelian randomization (MR) analysis to examine the causal effects of plasma proteins on CRC. Mediation analysis was performed to assess the indirect effects of plasma proteins on CRC through associated risk factors. In addition, we conducted a phenome-wide association study using the UK Biobank dataset to examine associations between these plasma proteins and other phenotypes. RESULTS: Out of 4,489 plasma proteins, MR analysis revealed causal associations with CRC for 23 proteins, including VIMP, MICB, TNFRSF11B, C5orf38 and SLC5A8. Our findings also confirm the associations between reported risk factors and CRC. Mediation analysis identified mediating effects of proteins on CRC outcomes through risk factors. Furthermore, MR analysis identified 154 plasma proteins are causally linked to at least one CRC risk factor. CONCLUSIONS: Our study evaluated the causal relationships between plasma proteins and CRC, providing a more complete understanding of potential therapeutic targets for CRC.
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Neoplasias Colorrectales , Proteoma , Humanos , Proteoma/genética , Análisis de la Aleatorización Mendeliana , Factores de Riesgo , Proteínas Sanguíneas , Estudio de Asociación del Genoma Completo , Polimorfismo de Nucleótido Simple , Transportadores de Ácidos MonocarboxílicosRESUMEN
BACKGROUND: Despite their high repair capability, bone defects still present a major challenge in orthopedic tissue engineering. Osteoblast differentiation is central to the treatment of bone defects. METHODS AND RESULTS: We used nasal mucosal-derived ectoderm mesenchymal stem cells (EMSCs) to promote osteogenic differentiation by co-culturing MC3T3-E1 cells. Our results showed that MC3T3-E1/EMSCs co-culture upregulated bone-related proteins and transglutaminase 2 (TG2) and increased alkaline phosphatase (ALP) activity and bone nodule formation relative to controls. Furthermore, our results showed that EMSC-derived sonic hedgehog (Shh) accounted for the enhanced MC3T3-E1 differentiation because inhibiting Shh signaling substantially reduced osteogenic differentiation. CONCLUSION: Altogether, these results suggest that EMSCs differentiated into osteoblast cells and supported MC3T3-E1 differentiation. Thus, EMSCs may be a promising cell source for treating bone-related diseases.
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Proteínas Hedgehog , Células Madre Mesenquimatosas , Proteínas Hedgehog/metabolismo , Osteogénesis , Ectodermo/metabolismo , Diferenciación Celular , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/metabolismoRESUMEN
A series of seven Cu/SSZ-13 catalysts with Si/Al = 6.7 are used to elucidate key rate-controlling factors during low-temperature standard ammonia-selective catalytic reduction (NH3-SCR), via a combination of SCR kinetics and operando electron paramagnetic resonance (EPR) spectroscopy. Strong Cu-loading-dependent kinetics, with Cu atomic efficiency increasing nearly by an order of magnitude, is found when per chabazite cage occupancy for Cu ion increases from â¼0.04 to â¼0.3. This is due mainly to the release of intercage Cu transfer constraints that facilitates the redox chemistry, as evidenced from detailed Arrhenius analysis. Operando EPR spectroscopy studies reveal strong connectivity between Cu-ion dynamics and SCR kinetics, based on which it is concluded that under low-temperature steady-state SCR, kinetically most relevant Cu species are those with the highest intercage mobility. Transient binuclear Cu species are mechanistically relevant species, but their splitting and cohabitation are indispensable for low-temperature kinetics.
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Amoníaco , Cobre , Amoníaco/química , Cobre/química , Espectroscopía de Resonancia por Spin del Electrón , Cinética , TemperaturaRESUMEN
BACKGROUND: Neonatal hypoxic-ischemic brain injury (HIE) is caused by perinatal asphyxia, which is associated with various confounding factors. Although studies on the pathogenesis and treatment of HIE have matured, sub-hypothermia is the only clinical treatment available for HIE. Previous evidence indicates that chlorogenic acid (CGA) exerts a potential neuroprotective effect on brain injury. However, the role of CGA on neonatal HI brain damage and the exact mechanism remains elusive. Here, we investigate the effects of CGA on HI models in vivo and in vitro and explore the underlying mechanism. METHODS: In the in vivo experiment, we ligated the left common carotid artery of 7-day-old rats and placed the rats in a hypoxic box for 2 h. We did not ligate the common carotid artery of the pups in the sham group since they did not have hypoxia. Brain atrophy and infarct size were evaluated by Nissl staining, HE staining and 2,3,5-triphenyltetrazolium chloride monohydrate (TTC) staining. Morris Water Maze test (MWM) was used to evaluate neurobehavioral disorders. Western-blotting and immunofluorescence were used to detect the cell signaling pathway. Malondialdehyde (MDA) content test, catalase (CAT) activity detection and Elisa Assay was used to detect levels of inflammation and oxidative stress. in vitro experiments were performed on isolated primary neurons. RESULT: In our study, pretreatment with CGA significantly decreased the infarct volume of neonatal rats after HI, alleviated brain edema, and improved tissue structure in vivo. Moreover, we used the Morris water maze to verify CGA's effects on enhancing the learning and cognitive ability and helping to maintain the long-term spatial memory after HI injury. However, Sirt1 inhibitor EX-527 partially reversed these therapeutic effects. CGA pretreatment inhibited neuronal apoptosis induced by HI by reducing inflammation and oxidative stress. The findings suggest that CGA potentially activates Sirt1 to regulate the Nrf2-NF-κB signaling pathway by forming complexes thereby protecting primary neurons from oxygen-glucose deprivation (OGD) damage. Also, CGA treatment significantly suppresses HI-induced proliferation of glial. CONCLUSION: Collectively, this study uncovered the underlying mechanism of CGA on neonatal HI brain damage. CGA holds promise as an effective neuroprotective agent to promote neonatal brain recovery from HI-induced injury. Video Abstract.
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Lesiones Encefálicas , Hipoxia-Isquemia Encefálica , Fármacos Neuroprotectores , Animales , Animales Recién Nacidos , Encéfalo/metabolismo , Lesiones Encefálicas/tratamiento farmacológico , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/patología , Ácido Clorogénico/metabolismo , Ácido Clorogénico/farmacología , Ácido Clorogénico/uso terapéutico , Hipoxia-Isquemia Encefálica/complicaciones , Hipoxia-Isquemia Encefálica/tratamiento farmacológico , Hipoxia-Isquemia Encefálica/metabolismo , Infarto/tratamiento farmacológico , Infarto/metabolismo , Infarto/patología , Inflamación/metabolismo , Isquemia/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , FN-kappa B/metabolismo , Fármacos Neuroprotectores/metabolismo , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Sirtuina 1/metabolismoRESUMEN
BACKGROUND: Embryos with higher morphologic quality grading may have a greater potential to achieve clinical pregnancy that leads to a live birth regardless of the type of cleavage-stage embryos or blastocysts. Few studies have investigated the impacts of embryo grading on the long-term health of the offspring. OBJECTIVE: This pilot study aimed to examine the associations between embryo morphologic quality and the physical, metabolic, and cognitive development of singletons conceived by in vitro fertilization and intracytoplasmic sperm injection at preschool age. STUDY DESIGN: This matched cohort study included singletons born to infertile couples who underwent fresh cleavage-stage embryo transfer cycles with good- or poor-quality embryos from 2014 to 2016 at the reproductive center of the Women's Hospital, School of Medicine, Zhejiang University. A total of 144 children, aged 4 to 6 years, participated in the follow-up assessment from 2020 to 2021, and the response rate of poor-quality embryo offspring was 39%. Singletons in the good-quality embryo group were matched with singletons in the poor-quality embryo group at a 2:1 ratio according to the fertilization method and the children's age (±1 year). We measured the offspring's height, weight, body mass index, blood pressure, thyroid hormone levels, and metabolic indicators. Neurodevelopmental assessments were performed using the Chinese version of the Wechsler Preschool and Primary Scale of Intelligence, Fourth Edition, and the Adaptive Behavior Assessment System, Second Edition. We also collected data from the medical records. A linear regression model was used to analyze the association between embryo morphologic quality and offspring health outcomes. RESULTS: A total of 48 singletons conceived with poor-quality embryo transfer and 96 matched singletons conceived with good-quality embryo transfer were included in the final analysis. Age, sex, height, weight, body mass index, blood pressure, thyroid function, and metabolic indicators were comparable between the 2 groups. After adjustment for potential risk factors by linear regression model 1 and model 2, poor-quality embryo offspring exhibited a tendency toward higher free thyroxine levels than offspring of good-quality embryo transfers (beta, 0.22; 95% confidence interval, 0.09-0.90; beta, 0.22; 95% confidence interval, 0.09-0.91, respectively), but this difference was not clinically significant. Regarding neurodevelopmental assessments, there was no difference in the full-scale intelligence quotient based on the Wechsler Preschool and Primary Scale of Intelligence (109.96±12.42 vs 109.60±14.46; P=.88) or the general adaptive index based on the Adaptive Behavior Assessment System (108.26±11.70 vs 108.08±13.44; P=.94) between the 2 groups. The subindices of the 2 tests were also comparable. These findings remained after linear regression analysis. CONCLUSION: At 4 to 6 years of age, singletons born from poor-quality embryo transfers have comparable metabolic and cognitive development as those born from good-quality embryo transfers using fresh cleavage-stage embryos. The results of this pilot study indicate that poor-quality embryos that can survive implantation and end in live birth are likely to have a developmental potential comparable to that of good-quality embryos.
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Semen , Inyecciones de Esperma Intracitoplasmáticas , Niño , Preescolar , Cognición , Estudios de Cohortes , Femenino , Fertilización , Fertilización In Vitro/efectos adversos , Humanos , Masculino , Proyectos Piloto , Embarazo , Inyecciones de Esperma Intracitoplasmáticas/efectos adversosRESUMEN
Heterotrimeric G proteins function as key players in guard cell signaling to many stimuli, including ultraviolet B (UV-B) and ethylene, but whether guard cell G protein signaling is activated by the only one potential G protein-coupled receptor, GCR1, is still unclear. Here, we found that gcr1 null mutants showed defects in UV-B- and ethylene-induced stomatal closure and production of reactive oxygen species (ROS) and nitric oxide (NO) in guard cells, but these defects could be rescued by the application of a Gα activator or overexpression of a constitutively active form of Gα subunit GPA1 (cGPA1). Moreover, the exogenous application of hydrogen peroxide (H2O2) or NO triggered stomatal closure in gcr1 mutants and cGPA1 transgenic plants in the absence or presence of UV-B or ethylene, but exogenous ethylene could not rescue the defect of gcr1 mutants in UV-B-induced stomatal closure, and gcr1 mutants did not affect UV-B-induced ethylene production in Arabidopsis leaves. These results indicate that GCR1 positively controls UV-B- and ethylene-induced stomatal closure by activating GPA1-dependent ROS and NO production in guard cells and that ethylene acts upstream of GCR1 to transduce UV-B guard cell signaling, which establishes the existence of a classic paradigm of G protein signaling in guard cell signaling to UV-B and ethylene.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Etilenos/metabolismo , Etilenos/farmacología , Subunidades alfa de la Proteína de Unión al GTP/metabolismo , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , Óxido Nítrico/metabolismo , Estomas de Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
BACKGROUND: There has been increasing interest in the relationship between body mass index(BMI) and pregnancy outcomes, especially in women undergoing frozen embryo transfer(FET). Several observational studies have been published, but so far with conflicting results. METHODS: A systematic review and meta-analysis was conducted according to PRISMA guidelines. Pubmed, Embase, Cochrane Library, Clinicaltrails.gov and Web of Science databases were searched based on established search strategy from inception through January 2021. RESULTS: Twelve studies were eligible. In women following FET, high BMI (BMI ≥ 23 kg/m2) was associated with an impaired live birth rate (LBR, OR: 0.89, 95% CI: 0.82-0.96, P = 0.002), but wasn't associated with the implantation rate or the clinical pregnancy rate. Subgroup analysis revealed higher LBR for women didn't complicated by polycystic ovary syndrome (PCOS, OR: 0.96, 95% CI: 0.85-1.08, P = 0.46) and women with blastocyst transferred (OR: 0.89, 95% CI: 0.68-1.16, P = 0.40). LBR did not differ between the low BMI group (BMI < 18.5 kg/m2) and the normal weight group. CONCLUSIONS: Our study showed that high BMI in women is negatively associated with LBR in FET cycles, whereas low BMI isn't. The results of subgroup analysis implied a need for women with a high BMI to get individualized weight management and treatment. Further evidence is still required to optimize preconception health and develop Nutritional and exercise guidelines.
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Índice de Masa Corporal , Criopreservación/métodos , Transferencia de Embrión/métodos , Resultado del Embarazo , Adulto , Tasa de Natalidad , Femenino , Humanos , Embarazo , Índice de EmbarazoRESUMEN
In this study, we developed a superstrong and reversible adhesive, which can possess a high bonding strength in the "adhesive" state and detach with the application of heating. An ionic crystal (IC) gel, in which an IC was immobilized within a soft-polymer matrix, were synthesized via in situ photo-crosslinking of a precursor solution composed of N, N-dimethyl acrylamide (DMAA) and a melted IC. The obtained IC gel is homogenous and transparent at melt point. When cooled to the phase transition temperature of the IC, the gel turns into the adhesive with the adhesion strength of 5.82â MPa (on glasses), due to the excellent wetting of melted gel and a thin layer of crystalline IC with high cohesive strength formed on the substrates. The synergistic effects between IC, polymer networks and substrates were investigated by solid state 1 Hâ NMR and molecular dynamics simulation. Such an adhesive layer is reversable and can be detached by heating and subsequent re-adhesion via cooling. This study proposed the new design of removable adhesives, which can be used in dynamic and complex environments.
RESUMEN
Primary ovarian insufficiency (POI) leads to infertility and premature menopause in young women. The genetic etiology of this disorder remains unknown in most patients. Using whole exome sequencing of a large Chinese POI pedigree, we identified a heterozygous 5 bp deletion inducing a frameshift in BNC1, which is predicted to result in a non-sense-mediated decay or a truncated BNC1 protein. Sanger sequencing identified another BNC1 missense mutation in 4 of 82 idiopathic patients with POI, and the mutation was absent in 332 healthy controls. Transfection of recombinant plasmids with the frameshift mutant and separately with the missense mutant in HEK293T cells led to abnormal nuclear localization. Knockdown of BNC1 was found to reduce BMP15 and p-AKT levels and to inhibit meiosis in oocytes. A female mouse model of the human Bnc1 frameshift mutation exhibited infertility, significantly increased serum follicle-stimulating hormone, decreased ovary size and reduced follicle numbers, consistent with POI. We report haploinsufficiency of BNC1 as an etiology of human autosomal dominant POI.
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Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Mutación Missense , Insuficiencia Ovárica Primaria/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Adulto , Anciano , Animales , Pueblo Asiatico/genética , Análisis Mutacional de ADN , Modelos Animales de Enfermedad , Femenino , Células HEK293 , Humanos , Ratones , Ratones Transgénicos , Persona de Mediana Edad , Linaje , Insuficiencia Ovárica Primaria/metabolismo , Secuenciación del Exoma , Adulto JovenRESUMEN
BACKGROUND: Endometrial injury can result in thin endometrium and subfertility. Granulocyte macrophage colony stimulating factor (GM-CSF) contributes to tissue repair, but its role in endometrial regeneration has not been investigated. METHODS: To determine the effect of GM-CSF on endometrial regeneration, we established a mouse model of thin endometrium by uterine perfusion with 20⯵L 90% ethanol. Thin endometrium in mice was featured by lowered endometrial thickness, decreased expression of Ki67 in glandular cells, and a reduced number of implantation sites. To explore the mechanism of GM-CSF on endometrial regeneration, endometrium was obtained from patients undergoing hysterectomy or hysteroscopy and endometrial biopsy. Effects of GM-CSF on primary cultured human endometrial glandular and stromal cells were examined by the 5-bromo-2'-deoxyuridine (BrdU) proliferation assay and transwell migration assay, followed by exploration of the potential signaling pathway. RESULTS: GM-CSF intraperitoneal (i.p.) injection significantly increased endometrial thickness, expression of Ki67 in endometrial glandular cells, and the number of implantation sites. GM-CSF significantly promoted proliferation of primary human endometrial glandular cells and migration of stromal cells. GM-CSF activated p-Akt and increased expressions of p70S6K and c-Jun, which were blocked by LY294002. CONCLUSION: We found that GM-CSF could improve endometrial regeneration, possibly through activating PI3K/Akt signaling pathway.
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Endometrio/efectos de los fármacos , Endometrio/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/sangre , Regeneración/efectos de los fármacos , Adulto , Animales , Biopsia , Bromodesoxiuridina/farmacología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Cromonas/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Histerectomía , Inyecciones Intraperitoneales , Quinasas Janus/metabolismo , Antígeno Ki-67/metabolismo , Ratones , Ratones Endogámicos ICR , Persona de Mediana Edad , Morfolinas/farmacología , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismoRESUMEN
Our previous work has demonstrated the high efficiency of CD8+ natural killer T (NKT)-like cells in killing antigen-bearing dendritic cells. To evaluate their role in the tumor microenvironment, we performed in vitro and in vivo antitumor experiments to investigate whether CD8+NKT-like cells could kill Yac-1 and B16 cells like NK cells and kill EL4-OVA8 cells in an antigen-specific manner like cytotoxic T lymphocytes (CTLs). Unlike NK1.1-CTLs, CD8+NKT-like cells also exhibit the capability to kill myeloid-derived suppressor cells (MDSCs) in an antigen-specific manner, indicative of their potential role in clearing tumor antigen-bearing MDSCs to improve the antitumor microenvironment. In vitro blocking experiments showed that granzyme B inhibitor efficiently suppressed the cytotoxicity of CD8+NKT-like cells against tumor cells and MDSCs, while Fas ligand (FasL) or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) inhibition failed to produce similar effects. Transcriptomic and phenotypic analyses of CD8+NKT-like cells, NK cells, and NK1.1-CTLs indicated that CD8+NKT-like cells expressed both T-cell activation markers and NK cell markers, thus bearing features of both the activated T cells and NK cells. Taken together, CD8+NKT-like cells could exert NK- and CTL-like antitumor effects through the elimination of both tumor cells and MDSCs in a granzyme B-dependent manner.
Asunto(s)
Células Asesinas Naturales/inmunología , Células Supresoras de Origen Mieloide/inmunología , Células T Asesinas Naturales/inmunología , Neoplasias Experimentales/terapia , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos CD8/metabolismo , Citotoxicidad Inmunológica , Femenino , Granzimas/metabolismo , Humanos , Activación de Linfocitos , Melanoma Experimental , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Neoplasias Experimentales/inmunología , Transcriptoma , Microambiente TumoralRESUMEN
OBJECTIVE: Any benefit of oestradiol supplementation with progesterone for luteal support after fresh embryo transfer in in vitro fertilisation/intracytoplasmic sperm injection (IVF/ICSI) cycles remains controversial. In this study, we further addressed this question in cycles using gonadotropin-releasing hormone (GnRH) agonist for ovarian stimulation. DESIGN: A retrospective cohort study in a tertiary teaching and research hospital. PATIENT(S): A total of 1602 patients were given oestradiol valerate (E) in addition to progesterone (P) as luteal support. One thousand six hundred and two patients receiving progesterone alone were selected as the control group. MAIN OUTCOME MEASURE(S): Live birth rate. Secondary measures included clinical pregnancy rate, miscarriage rate and premature birth rate. RESULT(S): Clinical pregnancy and live birth rates were similar for the P alone vs the P+E group. In cycles with oestradiol (E2) levels less than 5000 pmol/L on the day of hCG trigger, E supplementation resulted in a significantly higher live birth rate (23.44% vs 32.92%, OR = 1.60 [95% CI 1.05 to 2.46]). In cycles with oestradiol levels 5000 to 10 000 pmol/L on the day of hCG trigger, E supplementation did not increase the live birth rate (34.43% vs 35.42%, OR = 0.90 [95% CI 0.80 to 1.01]). In cycles with oestradiol levels over 10 000 pmol/L on the day of hCG trigger, the live birth rate was significantly lower (36.83% vs 31.37%, OR = 0.78 [95% CI 0.62 to 0.99]) and the premature birth rate was significantly higher (19.66% vs 28.73%,OR = 1.65 [95% CI 1.05 to 2.59]) in the E supplementation group. CONCLUSION(S): Any benefit of oestradiol supplementation for luteal phase support appears to correlate with the serum oestradiol level on the day of hCG trigger. Oestradiol supplementation is beneficial for improving live birth rate in cycles with oestradiol levels less than 5000 pmol/L, but is not recommended in cycles with oestradiol levels over 10 000 pmol/L.
RESUMEN
PURPOSE: The occurrence of diminished ovarian reserve (DOR) in women was growing in recent years. Although in vitro fertilization and embryo transfer (IVF-ET) became an effective treatment for DOR, the live-birth (LB) rate remains unsatisfactory. This study aimed to investigate the impact factors of LB rate in women with DOR undergoing assisted reproduction. METHODS: This was a single-center retrospective cohort study. A total of 2277 IVF-ET or ICSI cycles from 1957 DOR women were analysed. Impact factors of LB rate were explored via Student's t test, Pearson's Chi-square test, and multivariate logistic regression models. RESULTS: There were statistically significant differences in maternal age (P < 0.001), duration of infertility (P < 0.001), female body mass index (P = 0.039), first IVF cycle (P = 0.004), poor ovarian response (P < 0.001), paternal age (P < 0.001), total gonadotropin dose (P = 0.010), endometrial thickness (P = 0.021), number of follicles ≥ 14 mm (P = 0.007), number of oocytes retrieved (P < 0.001), number of frozen embryos (P = 0.014), and the stage (P < 0.001) and number (P < 0.001) of embryos transferred between the non-live-birth (NLB) and LB groups. However, only factors of maternal age, the stage and number of embryos transferred remained different after adjusting for potential confounders. CONCLUSIONS: Maternal age, the stage and number of embryos transferred were independent impact factors affecting the live-birth rate in women with DOR seeking for assisted conception.
Asunto(s)
Transferencia de Embrión/métodos , Fertilización In Vitro/métodos , Nacimiento Vivo , Reserva Ovárica , Resultado del Embarazo , Adulto , Tasa de Natalidad , Estudios de Cohortes , Femenino , Humanos , Infertilidad/terapia , Edad Materna , Enfermedades del Ovario , Inducción de la Ovulación/métodos , Embarazo , Índice de Embarazo , Estudios Retrospectivos , Motilidad EspermáticaRESUMEN
Visible/near infrared spectroscopy combined with chemometrics methods was used to detect ternary system adulteration in camellia oil quantificationally. In order to get adulterated samples, rapeseed oil and peanut oil were added to pure camellia oil in different proportion. Visible/near infrared spectroscopy data of pure and adulterated camellia oil samples were acquired in the wavelength range of 350~1800nm, and samples were randomly divided into calibration set and prediction set. The adulteration models were optimized by comparing different wavelength ranges, pretreatment methods and calibration methods The results show that the optimal modeling wavelength ranges and pretreatment methods for the prediction models of rapeseed oil, peanut oil and total adulteration amount are 750~1 770, 900~1 770, 870~1 770 nm and Multiple scattering correction (MSC), Standard normal variate (SNV) and second order differentia, and the best modeling method is Least square support vector machine (LSSVM). The correlation coefficient (R(P)) in prediction set and the root mean square error predictions(RMSEPs) of optimal adulteration models for rapeseed oil, peanut oil and total adulteration are 0.963, 0.982, 0.993 and 2.1%, 1.5%, 1.8%, respectively. Thus it can be seen that visible /near infrared spectroscopy combined with chemometrics methods can be used for quantitative ternary system adulteration detection in camellia oil.
Asunto(s)
Camellia , Contaminación de Alimentos , Espectroscopía Infrarroja Corta , Calibración , Análisis de los Mínimos Cuadrados , Aceites de Plantas , Máquina de Vectores de SoporteRESUMEN
LIBS (laser-induced breakdown spectroscopy) was used to detect Fe element content in soybean oil quantitatively. In this experiment, a series of soybean oil samples with different concentrations of Fe were used; LIBS spectra were collected with a two-channel high precision spectrometer. According to the LIBS spectrum of samples, two characteristic wavelength of Fe (404.58 and 406.36 nm) were determined, and different simple regression methods (exponential regression, linear regression and quadratic regression) were used to establish the quantitative analysis models of Fe content using each characteristic spectral line. The results indicate that the average relative error of Fe I 404.58 and Fe I 406.36 in simple exponential regression, linear regression and quadratic regression models were 29.49%, 8.93%, 8.70% and 28.95%, 8.63%, 8.44%, respectively. The results of Fe I 406.36 regression models is better than that of Fe I 404.58, and the quadratic regression model is optimal among the three regression models. According to these results, LIBS technology has certain feasibility for detecting Fe in soybean oil; the quadratic linear regression model can improve the prediction accuracy of Fe element effectively.