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It remains to be determined whether there is a geographical distribution pattern and phylogenetic signals for the Mycena strains with seed germination of the orchid plant Gastrodia elata. This study analyzed the community composition and phylogenetics of 72 Mycena strains associated with G. elata varieties (G. elata. f. glauca and G. elata. f. viridis) using multiple gene fragments (ITS+nLSU+SSU). We found that (1) these diverse Mycena phylogenetically belong to the Basidiospore amyloid group. (2) There is a phylogenetic signal of Mycena for germination of G. elata. Those strains phylogenetically close to M. abramsii, M. polygramma, and an unclassified Mycena had significantly higher germination rates than those to M. citrinomarginata. (3) The Mycena distribution depends on geographic site and G. elata variety. Both unclassified Mycena group 1 and the M. abramsii group were dominant for the two varieties of G. elata; in contrast, the M. citrinomarginata group was dominant in G. elata f. glauca but absent in G. elata f. viridis. Our results indicate that the community composition of numerous Mycena resources in the Zhaotong area varies by geographical location and G. elata variety. Importantly, our results also indicate that Mycena's phylogenetic status is correlated with its germination rate.
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Gastrodia , Germinación , Filogenia , Gastrodia/microbiología , Gastrodia/genética , ADN de Hongos/genética , Semillas/microbiología , Semillas/crecimiento & desarrollo , Basidiomycota/genética , Basidiomycota/clasificación , Basidiomycota/fisiologíaRESUMEN
Protein retention expansion microscopy (ExM) retains genetically encoded fluorescent proteins or antibody-conjugated fluorescent probes in fixed tissue and isotropically expands the tissue through a swellable polymer network to allow nanoscale (<70 nm) resolution on diffraction-limited confocal microscopes. Despite numerous advantages ExM brings to biological studies, the full protocol is time-consuming and can take multiple days to complete. Here, we adapted the ExM protocol to the vibratome-sectioned brain tissue of Xenopus laevis tadpoles and implemented a microwave-assisted protocol to reduce the workflow from days to hours. In addition to the significantly accelerated processing time, our microwave-assisted ExM (M/WExM) protocol maintains the superior resolution and signal-to-noise ratio of the original ExM protocol. Furthermore, the M/WExM protocol yields higher magnitude of expansion, suggesting that in addition to accelerating the process through increased diffusion rate of reagents, microwave radiation may also facilitate the expansion process. To demonstrate the applicability of this method to other specimens and protocols, we adapted the microwave-accelerated protocol to whole mount adult brain tissue of Drosophila melanogaster fruit flies, and successfully reduced the total processing time of a widely-used Drosophila IHC-ExM protocol from 6 days to 2 days. Our results demonstrate that with appropriate adjustment of the microwave parameters (wattage, pulse duration, interval, and number of cycles), this protocol can be readily adapted to different model organisms and tissue types to greatly increase the efficiency of ExM experiments.
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Full myco-heterotrophic orchid Gastrodia elata Bl. is widely distributed in Northeast Asia, and previous research has not fully investigated the symbiotic fungal community of its early immature tubers. This study utilized Illumina sequencing to compare symbiotic fungal communities in natural G. elata immature tubers and their habitats. LEfSe (Linear Discriminant Analysis Effect Size) was used to screen for Biomarkers that could explain variations among different fungal communities, and correlation analyses were performed among Biomarkers and other common orchid mycorrhizal fungi. Our results illustrate that the symbiotic fungal communities of immature G. elata tubers cannot be simply interpreted as subsets of the environmental fungal communities because some key members cannot be traced back to the environment. The early growth of G. elata was related to a small group of fungi, such as Sebacina, Thelephora, and Inocybe, which were also common mycorrhizal fungi from other orchids. In addition, Mycena, Auricularia, and Cryptococcus were unique fungal partners of G. elata, and many new species have yet to be discovered. Possible symbiotic Mycena should be M. plumipes and its sibling species in this case. Our results provide insight into the symbiotic partner switch and trophic pattern change during the development and maturation of G. elata.
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In developing Xenopus tadpoles, the optic tectum begins to receive patterned visual input while visuomotor circuits are still undergoing neurogenesis and circuit assembly. This visual input regulates neural progenitor cell fate decisions such that maintaining tadpoles in the dark increases proliferation, expanding the progenitor pool, while visual stimulation promotes neuronal differentiation. To identify regulators of activity-dependent neural progenitor cell fate, we profiled the transcriptomes of proliferating neural progenitor cells and newly differentiated neurons using RNA-Seq. We used advanced bioinformatic analysis of 1,130 differentially expressed transcripts to identify six differentially regulated transcriptional regulators, including Breast Cancer 1 (BRCA1) and the ETS-family transcription factor, ELK-1, which are predicted to regulate the majority of the other differentially expressed transcripts. BRCA1 is known for its role in cancers, but relatively little is known about its potential role in regulating neural progenitor cell fate. ELK-1 is a multifunctional transcription factor which regulates immediate early gene expression. We investigated the potential functions of BRCA1 and ELK-1 in activity-regulated neurogenesis in the tadpole visual system using in vivo time-lapse imaging to monitor the fate of GFP-expressing SOX2+ neural progenitor cells in the optic tectum. Our longitudinal in vivo imaging analysis showed that knockdown of either BRCA1 or ELK-1 altered the fates of neural progenitor cells and furthermore that the effects of visual experience on neurogenesis depend on BRCA1 and ELK-1 expression. These studies provide insight into the potential mechanisms by which neural activity affects neural progenitor cell fate.
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Células-Madre Neurales , Colículos Superiores , Animales , Genes BRCA1 , Neuronas , Proteínas Proto-Oncogénicas c-ets , Xenopus laevis/genética , Proteína Elk-1 con Dominio ets , Proteína BRCA1RESUMEN
BigNeuron is an open community bench-testing platform with the goal of setting open standards for accurate and fast automatic neuron tracing. We gathered a diverse set of image volumes across several species that is representative of the data obtained in many neuroscience laboratories interested in neuron tracing. Here, we report generated gold standard manual annotations for a subset of the available imaging datasets and quantified tracing quality for 35 automatic tracing algorithms. The goal of generating such a hand-curated diverse dataset is to advance the development of tracing algorithms and enable generalizable benchmarking. Together with image quality features, we pooled the data in an interactive web application that enables users and developers to perform principal component analysis, t-distributed stochastic neighbor embedding, correlation and clustering, visualization of imaging and tracing data, and benchmarking of automatic tracing algorithms in user-defined data subsets. The image quality metrics explain most of the variance in the data, followed by neuromorphological features related to neuron size. We observed that diverse algorithms can provide complementary information to obtain accurate results and developed a method to iteratively combine methods and generate consensus reconstructions. The consensus trees obtained provide estimates of the neuron structure ground truth that typically outperform single algorithms in noisy datasets. However, specific algorithms may outperform the consensus tree strategy in specific imaging conditions. Finally, to aid users in predicting the most accurate automatic tracing results without manual annotations for comparison, we used support vector machine regression to predict reconstruction quality given an image volume and a set of automatic tracings.
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Benchmarking , Microscopía , Microscopía/métodos , Imagenología Tridimensional/métodos , Neuronas/fisiología , AlgoritmosRESUMEN
OBJECTIVES: To study the moderating effect of mother-child relationship in the association between maternal parenting stress and emotional and behavioral problems in preschool children, and to provide reference for the prevention and control of emotional and behavioral problems in preschool children. METHODS: Using a stratified cluster sampling method, 2 049 preschool children were surveyed from November to December 2021, who sampled from 12 kindergartens in Wuhu City, Anhui Province. The emotional and behavioral problems of preschool children were assessed with the Strength and Difficulties Questionnaire. Pearson correlation analysis was used to evaluate the relationship of maternal parenting stress and mother-child relationship with children's emotional and behavioral problems. The PROCESS Macro was used to analyze the moderating effect of conflicted and dependent mother-child relationships in the association between maternal parenting stress and emotional and behavioral problems in these preschool children. RESULTS: Among these preschool children, maternal parenting stress was positively correlated with the scores of emotional symptoms, conduct problems, hyperactivity, and peer problems subscales and total difficulty scores (P<0.001); intimate mother-child relationships were negatively correlated with the scores of conduct problems, hyperactivity, and peer problems subscales and total difficulty scores (P<0.001); conflicted and dependent mother-child relationships were positively correlated with the scores of emotional symptoms, conduct problems, hyperactivity, and peer problems subscales and total difficulty scores (P<0.001). After controlling for relevant confounding factors, conflicted mother-child relationship (ß=0.05, P=0.001) and dependent mother-child relationship (ß=0.04, P=0.012) were found to have a moderating effect on the association between maternal parenting stress and total difficulty scores in these preschool children. CONCLUSIONS: Negative mother-child relationships play a moderating role in the association between maternal parenting stress and emotional and behavioral problems in preschool children. Prevention of emotional and behavioral problems in preschool children should focus on reducing maternal parenting stress and improving negative mother-child relationships.
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Problema de Conducta , Humanos , Preescolar , Femenino , Problema de Conducta/psicología , Responsabilidad Parental/psicología , Emociones , Relaciones Madre-Hijo , Encuestas y Cuestionarios , Madres/psicologíaRESUMEN
Bacterial azapteridine-containing phytotoxin toxoflavin is a causal agent of rice grain rot. Here, we heterologously reconstitute Bukholderia toxoflavin biosynthesis in Escherichia coli and identify key pathway intermediates, including the hitherto unknown ribityl-dedimethyl-toxoflavin. Furthermore, we characterized a cofactorless oxidase that converts ribityl-dedimethyl-toxoflavin to ribose and dedimethyl-toxoflavin, the latter of which then undergoes stepwise methylations to form toxoflavin. These findings provide new insights into the biosynthetic pathways of toxoflavin and related triazine metabolites.
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Oxidorreductasas , Pirimidinonas , Triazinas , Escherichia coli/genética , Escherichia coli/metabolismoRESUMEN
Protein degradation is critical for brain function through processes that remain incompletely understood. Here, we investigated the in vivo function of the 20S neuronal membrane proteasome (NMP) in the brain of Xenopus laevis tadpoles. With biochemistry, immunohistochemistry, and electron microscopy, we demonstrated that NMPs are conserved in the tadpole brain and preferentially degrade neuronal activity-induced newly synthesized proteins in vivo. Using in vivo calcium imaging in the optic tectum, we showed that acute NMP inhibition rapidly increased spontaneous neuronal activity, resulting in hypersynchronization across tectal neurons. At the circuit level, inhibiting NMPs abolished learning-dependent improvement in visuomotor behavior in live animals and caused a significant deterioration in basal behavioral performance following visual training with enhanced visual experience. Our data provide in vivo characterization of NMP functions in the vertebrate nervous system and suggest that NMP-mediated degradation of activity-induced nascent proteins may serve as a homeostatic modulatory mechanism in neurons that is critical for regulating neuronal activity and experience-dependent circuit plasticity.
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Neuronas , Complejo de la Endopetidasa Proteasomal , Animales , Complejo de la Endopetidasa Proteasomal/metabolismo , Neuronas/metabolismo , Colículos Superiores/fisiología , Techo del Mesencéfalo , Xenopus laevis/metabolismo , Reacción de Prevención/fisiología , Larva/metabolismo , Plasticidad Neuronal/fisiologíaRESUMEN
BACKGROUND: New and more severe clinical manifestations associated with the coronavirus disease 2019 (COVID-19) are emerging constantly in the pediatric age group. Patients in this age group are also primary carriers of the influenza virus and are at a higher risk of developing severe infection. However, studies comparing influenza and COVID-19 to show which condition causes a more severe form of disease amongst the pediatric age group are scarce. AIM: To compare the laboratory results, clinical symptoms and clinical outcomes in pediatric patients with COVID-19 and influenza. METHODS: A systematic and comprehensive search was carried out in databases and search engines, including EMBASE, Cochrane, MEDLINE, ScienceDirect and Google Scholar from 1964 until January 2022. A meta-analysis was carried out using a random-effects model and pooled odds ratio (OR) or standardized mean difference (SMD) and 95%CI. RESULTS: A total of 16 studies satisfied the inclusion criteria. Pediatric COVID-19 patients had a significantly reduced risk of cough (pooled OR = 0.16; 95%CI: 0.09 to 0.27), fever (pooled OR = 0.23; 95%CI: 0.12 to 0.43), and dyspnea (pooled OR = 0.54; 95%CI: 0.33 to 0.88) compared to influenza patients. Furthermore, total hemoglobin levels (pooled SMD = 1.22; 95%CI: 0.29 to 2.14) in COVID-19 patients were significantly higher as compared to pediatric influenza patients. There was no significant difference in symptoms such as sore throat, white blood cell count, platelets, neutrophil and lymphocytes levels, and outcomes like mortality, intensive care unit admission, mechanical ventilation or length of hospital stay. CONCLUSION: COVID-19 is associated with a significantly lower rate of clinical symptoms and abnormal laboratory indexes compared to influenza in the pediatric age group. However, further longitudinal studies of the outcomes between influenza and COVID-19 pediatric patients are needed.
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OBJECTIVES: To investigate the sleep patterns and characteristics of infants and young children and the association between sleep patterns and breastfeeding. METHODS: A general information questionnaire, Brief Infant Sleep Questionnaire (BISQ), and a questionnaire on feeding were used to investigate the sleep quality and feeding patterns of 1 148 infants and young children aged 7-35 months. The K-means clustering method was used to identify sleep patterns and characteristics. A multivariate logistic regression analysis was used to investigate the association between sleep patterns and breastfeeding. RESULTS: Three typical sleep patterns were identified for the 1 148 infants and young children aged 7-35 months: early bedtime and long sleep time; short sleep latency and moderate sleep time; late bedtime, prolonged sleep latency, and insufficient sleep time. The third pattern showed sleep disorders. The multivariate logistic regression analysis showed that compared with formula feeding, exclusive breastfeeding within 6 months after birth reduced the risk of sleep disorder patterns by 69% (OR=0.31, 95%CI: 0.11-0.81). The risk of sleep disorder patterns was reduced by 40% (OR=0.60, 95%CI: 0.38-0.96) in the infants receiving breastfeeding for 4-6 months compared with those receiving breastfeeding for 1-3 months. CONCLUSIONS: There are different sleep patterns in infants and young children, and breastfeeding can reduce the development of sleep disorder patterns.
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Lactancia Materna , Trastornos del Sueño-Vigilia , Lactante , Niño , Femenino , Humanos , Preescolar , Encuestas y Cuestionarios , Sueño , Análisis por ConglomeradosRESUMEN
The nitrogen-nitrogen bond is a core feature of diverse functional groups like hydrazines, nitrosamines, diazos, and pyrazoles. Such functional groups are found in >300 known natural products. Such N-N bond-containing functional groups are also found in significant percentage of clinical drugs. Therefore, there is wide interest in synthetic and enzymatic methods to form nitrogen-nitrogen bonds. In this review, we summarize synthetic and biosynthetic approaches to diverse nitrogen-nitrogen-bond-containing functional groups, with a focus on biosynthetic pathways and enzymes.
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Productos Biológicos , Nitrógeno , Productos Biológicos/química , Vías Biosintéticas , Hidrazinas/química , Hidrazinas/metabolismo , Nitrógeno/químicaRESUMEN
Intercellular transfer of toxic proteins between neurons is thought to contribute to neurodegenerative disease, but whether direct interneuronal protein transfer occurs in the healthy brain is not clear. To assess the prevalence and identity of transferred proteins and the cellular specificity of transfer, we biotinylated retinal ganglion cell proteins in vivo and examined biotinylated proteins transported through the rodent visual circuit using microscopy, biochemistry, and mass spectrometry. Electron microscopy demonstrated preferential transfer of biotinylated proteins from retinogeniculate inputs to excitatory lateral geniculate nucleus (LGN) neurons compared with GABAergic neurons. An unbiased mass spectrometry-based screen identified â¼200 transneuronally transported proteins (TNTPs) isolated from the visual cortex. The majority of TNTPs are present in neuronal exosomes, and virally expressed TNTPs, including tau and ß-synuclein, were detected in isolated exosomes and postsynaptic neurons. Our data demonstrate transfer of diverse endogenous proteins between neurons in the healthy intact brain and suggest that TNTP transport may be mediated by exosomes.
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Comunicación Celular/fisiología , Exosomas/metabolismo , Neuronas/metabolismo , Corteza Visual/metabolismo , Animales , Técnicas de Trazados de Vías Neuroanatómicas , Proteómica , Ratas , Ratas Wistar , Vías Visuales/metabolismo , XenopusRESUMEN
In vivo time-lapse imaging of complete dendritic arbor structures in tectal neurons of Xenopus laevis tadpoles has served as a powerful in vivo model to study activity-dependent structural plasticity in the central nervous system during early development. In addition to quantitative analysis of gross arbor structure, dynamic analysis of the four-dimensional data offers particularly valuable insights into the structural changes occurring in subcellular domains over experience/development-driven structural plasticity events. Such analysis allows not only quantifiable characterization of branch additions and retractions with high temporal resolution but also identification of the loci of action. This allows for a better understanding of the spatiotemporal association of structural changes to functional relevance. Here we describe a protocol for in vivo time-lapse imaging of complete dendritic arbors from individual neurons in the brains of anesthetized tadpoles with two-photon microscopy and data analysis of the time series of 3D dendritic arbors. For data analysis, we focus on dynamic analysis of reconstructed neuronal filaments using a customized open source computer program we developed (4D SPA), which allows aligning and matching of 3D neuronal structures across different time points with greatly improved speed and reliability. File converters are provided to convert reconstructed filament files from commercial reconstruction software to be used in 4D SPA. The program and user manual are publicly accessible and operate through a graphical user interface on both Windows and Mac OSX.
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Plasticidad Neuronal , Colículos Superiores , Animales , Dendritas/fisiología , Larva/fisiología , Plasticidad Neuronal/fisiología , Reproducibilidad de los Resultados , Colículos Superiores/fisiología , Imagen de Lapso de Tiempo , Xenopus laevisRESUMEN
Cyberchondria is considered "the anxiety-amplifying effects of online health-related searches." During the COVID-19 pandemic, people are likely to search health-related information online for reassurance because of fear and related physical symptoms, while cyberchondria may be triggered due to the escalation of health anxiety, different online seeking behavior preference, information overload, and insufficient e-health literacy. This study aimed to investigate the status and influencing factors of cyberchondria in residents in China during the epidemic period of COVID-19. The participants were 674 community residents of Nanyang city surveyed from February 1 to 15, 2020. We administered online measures, including the Chinese Short Form of the Cyberchondria Severity Scale (C-CSS-12), Short Health Anxiety Inventory (SHAI), eHealth Literacy Scale (eHEALS), Patient Health Questionnaire-15 (PHQ-15), and COVID-19-related online information seeking behavior questionnaire. In our study, the average C-CSS-12 total score of residents was 30.65 ± 11.53 during the virus epidemic; 25% of participants scored 22 or below, 50% scored 23 to 38, and 21.9% scored 39 to 60. The SHAI total score (ß = 0.598 > 0, P < 0.001), the use of general search engines (ß = 1.867 > 0, P = 0.039), and searching for information on how to diagnose COVID-19 (ß = 2.280 > 0, P = 0.020) were independent risk factors for cyberchondria, while searching lasting less than 10 min each (ß = -2.992 < 0, P = 0.048), the use of traditional media digital platforms (ß = -1.650 < 0, P = 0.024) and professional medical communication platforms (ß = -4.189 < 0, P = 0.007) were independent protective factors. Our findings showed that nearly a quarter of the participants scored 39 or higher on the C-CSS-12 in Nanyang city during the pandemic, which should be taken seriously. Health anxiety and COVID-19-related online information seeking behavior including online duration, topics and choice on different information channels were important influencing factors of cyberchondria. These findings have implications for further research and clinical practice on cyberchondria in China.
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[This corrects the article DOI: 10.1039/D0SC05843B.].
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Indolmycin is an antibiotic from Streptomyces griseus ATCC 12648 with activity against Helicobacter pylori, Plasmodium falciparum, and methicillin-resistant Staphylococcus aureus. Here we describe the use of the indolmycin biosynthetic genes in E. coli to make indolmycenic acid, a chiral intermediate in indolmycin biosynthesis, which can then be converted to indolmycin through a three-step synthesis. To expand indolmycin structural diversity, we introduce a promiscuous tryptophanyl-tRNA synthetase gene (trpS) into our E. coli production system and feed halogenated indoles to generate the corresponding indolmycenic acids, ultimately allowing us to access indolmycin derivatives through synthesis. Bioactivity testing against methicillin-resistant Staphylococcus aureus showed modest antibiotic activity for 5-, 6-, and 7-fluoro-indolmycin.
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Natural products with rare functional groups are likely to be constructed by unique biosynthetic enzymes. One such rare functional group is the O-methyl nitronate, which can undergo [3 + 2] cycloaddition reactions with olefins in mild conditions. O-methyl nitronates are found in some natural products; however, how such O-methyl nitronates are assembled biosynthetically is unknown. Here we show that the assembly of the O-methyl nitronate in the natural product enteromycin carboxamide occurs via activation of glycine on a peptidyl carrier protein, followed by reaction with a diiron oxygenase to give a nitronate intermediate and then with a methyltransferase to give an O-methyl nitronate. Guided by the discovery of this pathway, we then identify related cryptic biosynthetic gene cassettes in other bacteria and show that these alternative gene cassettes can, instead, facilitate oxidative denitrification of glycine-derived nitronates. Altogether, our work reveals bifurcating pathways from a central glycine-derived nitronate intermediate in bacteria.
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Proteínas Bacterianas/metabolismo , Glicina/química , Compuestos de Nitrógeno/química , Streptomyces , Proteínas Bacterianas/genética , Desnitrificación , Regulación Bacteriana de la Expresión Génica , MetilaciónRESUMEN
BACKGROUND: Angioimmunoblastic T cell lymphoma (AITL) is an infrequent hematological malignancy with variable and often atypical presentations. The presence of dysproteinemia, autoantibodies and systemic involvement in AITL has often led to a delay in diagnosis or even misdiagnosis in practice. We herewith present a case of AITL that primarily presented with acute kidney injury associated with type II Cryoglobulinemia, the underlying cause was only identified 8 months after the emergence of initial symptoms. CASE PRESENTATION: A 67-year old woman presented with 2-month history of intermittent joint pain and a 3-day history of bilateral lower limb edema and acute kidney injury. Initial laboratory investigations showed marked hypocomplementemia with positive autoantibodies of ANA, anti-cardiolipin-IgM and direct antiglobulin. The serum and urinary Immunofixation and serum cryoglobulin tests were negative, while the serum free κ to λ light chain ratio was 0.231. A renal biopsy showed a diffuse proliferative glomerulonephritis with intracapillary pseudothrombi formation. There were orderly arranged microtubular structures of 20-35 nm in diameter in the subendothelial and mesangial area on electron microscopy. Shortly afterwards, the patient developed tingling affecting her finger tips and weak hands and legs. A diagnosis of cryoglobulinemia complicated with cryoglobulinemic glomerulonephritis and polyneuropathy was made. She responded well to methylprednisolone, plasma exchange and rituximab. However, 3 months later, she presented with generalized pruritic rash, weight loss, and inguinal lymphadenopathy. A subsequent inguinal excisional lymph node biopsy at month 8 revealed AITL as the underlying disease. CONCLUSIONS: AITL and its associated B cell dysregulation can give rise to autoimmunity and cryoglobulinemia which may conceal itself as the underlying disorder. In various clinical scenarios of auto-immune diseases, it is advisable that the clinicians should take into consideration the multi-faceted lymphoma.
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Lesión Renal Aguda/etiología , Crioglobulinemia/etiología , Crioglobulinas/análisis , Diagnóstico Tardío , Linfoma de Células T/diagnóstico , Lesión Renal Aguda/patología , Adulto , Anciano , Análisis Químico de la Sangre , Complemento C3/análisis , Creatinina/sangre , Femenino , Humanos , Riñón/patología , Linfoma de Células T/sangre , Linfoma de Células T/complicaciones , Masculino , Persona de Mediana EdadRESUMEN
Background: Most organ donation coordinators suffer varying degrees of anxiety, depression and poor sleep caused by constant work pressure, and their professional identity is only at a medium level. All of this leads to a great risk of job burnout. Objective: To identify the influencing factors of and effective countermeasures against job burnout among organ donation coordinators. Method: Semistructured interviews were used for data collection. In-person or phone interviews were conducted from December 2017 to June 2018. Results: 12 organ donation coordinators who came from 7 different provinces and cities in China were interviewed. The interview data were sorted, and relevant topics were extracted and summarized in terms of two aspects, namely, factors that influenced job burnout in organ donation coordinators and effective countermeasures for dealing with job burnout. Conclusion: Factors influencing job burnout among organ donation coordinators include personal factors, job responsibilities, salary and benefit factors, and donor family factors. Measures to help organ donation coordinators effectively address burnout include self-regulation, social support, and positive events.