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Bacterial gene expression is a complex process involving extensive regulatory mechanisms. Along with growing interests in this field, Nanopore Direct RNA Sequencing (DRS) provides a promising platform for rapid and comprehensive characterization of bacterial RNA biology. However, the DRS of bacterial RNA is currently deficient in the yield of mRNA-mapping reads and has yet to be exploited for transcriptome-wide RNA modification mapping. Here, we showed that pre-processing of bacterial total RNA (size selection followed by ribosomal RNA depletion and polyadenylation) guaranteed high throughputs of sequencing data and considerably increased the amount of mRNA reads. This way, complex transcriptome architectures were reconstructed for Escherichia coli and Staphylococcus aureus and extended the boundaries of 225 known E. coli operons and 89 defined S. aureus operons. Utilizing unmodified in vitro-transcribed (IVT) RNA libraries as a negative control, several Nanopore-based computational tools globally detected putative modification sites in the E. coli and S. aureus transcriptomes. Combined with Next-Generation Sequencing-based N6-methyladenosine (m6A) detection methods, 75 high-confidence m6A candidates were identified in the E. coli protein-coding transcripts, while none were detected in S. aureus. Altogether, we demonstrated the potential of Nanopore DRS in systematic and convenient transcriptome and epitranscriptome analysis.
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Escherichia coli , Secuenciación de Nanoporos , ARN Bacteriano , Análisis de Secuencia de ARN , Staphylococcus aureus , Transcriptoma , Escherichia coli/genética , Escherichia coli/metabolismo , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Transcriptoma/genética , ARN Bacteriano/genética , ARN Bacteriano/metabolismo , Análisis de Secuencia de ARN/métodos , Secuenciación de Nanoporos/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Operón/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Adenosina/análogos & derivados , Adenosina/metabolismo , Adenosina/genética , Nanoporos , Regulación Bacteriana de la Expresión Génica , Perfilación de la Expresión Génica/métodosRESUMEN
The E11 cell line, derived from striped snakehead fish (Channa striata), possesses a distinctive feature: it is persistently infected with a C-type retrovirus. Notably, it exhibits high permissiveness to piscine nodavirus and the emerging tilapia lake virus (TiLV). Despite its popularity in TiLV research, the absence of genome assembly for the E11 cell line and Channa striata has constrained research on host-virus interactions. This study aimed to fill this gap by sequencing, assembling, and annotating the E11 cell line genome. Our efforts yielded a 600.5 Mb genome including 24 chromosomes with a BUSCO score of 98.8%. In addition, the complete proviral DNA sequence of snakehead retrovirus (SnRV) was identified in the E11 cell genome. Comparative genomic analysis between the E11 cell line and another snakehead species Channa argus revealed the loss of many immune-related gene families in the E11 cell genome, indicating a compromised immune response. We also conducted transcriptome analysis of mock- and TiLV-infected E11 cells, unveiling new perspectives on virus-virus and host-virus interactions. The TiLV infection suppressed the high expression of SnRV in E11 cells, and activated some other endogenous retroviruses. The protein-coding gene comparison revealed a pronounced up-regulation of genes involved in immune response, alongside a down-regulation of genes associated with specific metabolic processes. In summary, the genome assembly and annotation of the E11 cell line provide valuable resources to understand the SnRV and facilitate further studies on nodavirus and TiLV. The RNA-seq profiles shed light on the cellular mechanisms employed by fish cells in response to viral challenges, potentially guiding the development of therapeutic strategies against TiLV in aquaculture. This study also provides the first insights into the viral transcriptome profiles of endogenous SnRV and evading TiLV, enhancing our understanding of host-virus interactions in fish.
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Enfermedades de los Peces , Tilapia , Virus , Animales , Retroviridae , Cromosomas , Perfilación de la Expresión Génica/veterinariaRESUMEN
Neural tube defects (NTDs) represent a prevalent and severe category of congenital anomalies in humans. Cadmium (Cd) is an environmental teratogen known to cause fetal NTDs. However, its underlying mechanisms remain elusive. This study aims to investigate the therapeutic potential of lipophagy in the treatment of NTDs, providing valuable insights for future strategies targeting lipophagy activation as a means to mitigate NTDs.We successfully modeled NTDs by Cd exposure during pregnancy. RNA sequencing was employed to investigate the transcriptomic alterations and functional enrichment of differentially expressed genes in NTD placental tissues. Subsequently, pharmacological/genetic (Atg5-/- placentas) experiments confirmed that inducing placental lipophagy can alleviate Cd induced-NTDs. We found that Cd exposure caused NTDs. Further analyzed transcriptomic data from the placentas with NTDs which revealed significant downregulation of low-density lipoprotein receptor associated protein 1(Lrp1) gene expression responsible for positive regulation of low-density lipoprotein cholesterol (LDL-C) transport. Correspondingly, there was an increase in maternal serum/placenta/amniotic fluid LDL-C content. Subsequently, we have discovered that Cd exposure activated placental lipophagy. Pharmacological/genetic (Atg5-/- placentas) experiments confirmed that inducing placental lipophagy can alleviate Cd induced-NTDs. Furthermore, our findings demonstrate that activation of placental lipophagy effectively counteracts the Cd-induced elevation in LDL-C levels. Lipophagy serves to mitigate Cd-induced NTDs by reducing LDL-C levels within mouse placentas.
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Cadmio , LDL-Colesterol , Defectos del Tubo Neural , Placenta , Femenino , Animales , Embarazo , Placenta/metabolismo , Placenta/efectos de los fármacos , Defectos del Tubo Neural/genética , Defectos del Tubo Neural/inducido químicamente , Defectos del Tubo Neural/metabolismo , Ratones , Cadmio/toxicidad , LDL-Colesterol/sangre , Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/genética , Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
Diminished testosterone levels have been documented as a key factor in numerous male health disorders. Both human and animal studies have consistently demonstrated that cadmium (Cd), a pervasive environmental heavy metal, results in decreased testosterone levels. However, the exact mechanism through which Cd interferes with testosterone synthesis remains incompletely elucidated. This research sought to examine the impact of cellular senescence on Cd-suppressed testosterone synthesis. We also investigated the related m6A modification mechanism. The results demonstrated that Cd (100â¯mg/L) led to a decrease in testosterone levels, along with downregulated expression of testosterone synthase in C57BL/6â¯N male mice. Furthermore, Cd significantly increased ß-galactosidase staining intensity, senescence-related proteins, and senescence-related secretory phenotypes in mouse testicular Leydig cells. Subsequent investigations revealed that Cd decreased the mRNA and protein levels of NAD-dependent deacetylase Sirtuin-1 (SIRT1) in Leydig cells. Mechanistically, mice treated with resveratrol (50â¯mg/kg), a specific SIRT1 activator, mitigated Leydig cell senescence and reversed Cd-reduced testosterone levels in mouse testes. These effects were also restored by SIRT1 overexpression in Leydig cells. Additionally, we found that Cd increased the level of methyltransferase enzyme METTL3 and Sirt1 m6A modification in Leydig cells. Mettl3 siRNA effectively restored Cd-enhanced Sirt1 m6A level and reversed Cd-downregulated Sirt1 mRNA expression in Leydig cells. Overall, our findings suggest that Cd exposure inhibits testosterone synthesis via Sirt1 m6A modification-mediated senescence in mouse testes. These results offer an experimental basis for investigating the causes and potential treatments of hypotestosteronemia induced by environmental factors.
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This study aimed to assess PEMF in a rat model of senile osteoporosis and its relationship with NLRP3-mediated low-grade inflammation in the bone marrow microenvironment. A total of 24 Sprague Dawley (SD) rats were included in this study. Sixteen of them were 24-month natural-aged male SD rats, which were randomly distributed into the Aged group and the PEMF group (n = 8 per group). The remaining 8 3-month -old rats were used as the Young positive control group (n = 8). Rats in the PEMF group received 12 weeks of PEMF with 40 min/day, five days per week, while the other rats received placebo PEMF intervention. Bone mineral density/microarchitecture, serum levels of CTX-1 and P1CP, and NLRP3-related signaling genes and proteins in rat bone marrow were then analyzed. The 12-week of PEMF showed significant mitigation of aging-induced bone loss and bone microarchitecture deterioration, i.e. PEMF increased the bone mineral density of the proximal femur and L5 vertebral body and improved parameters of the proximal tibia and L4 vertebral body. Further analysis showed that PEMF reversed aging-induced bone turnover, specifically, decreased serum CTX-1 and elevated serum P1CP. Furthermore, PEMF also dramatically inhibited NLRP3-mediated low-grade inflammation in the bone marrow, i.e. PEMF inhibited the levels of NLRP3, proCaspase1, cleaved Caspase1, IL-1ß, and GSDMD-N. The study demonstrated that PEMF could mitigate the aging-induced bone loss and reverses the deterioration of bone microarchitecture probably through inhibiting NLRP3-mediated low-grade chronic inflammation to improve the inflammatory bone microenvironment in aged rats.
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Densidad Ósea , Campos Electromagnéticos , Inflamación , Proteína con Dominio Pirina 3 de la Familia NLR , Osteoporosis , Ratas Sprague-Dawley , Animales , Osteoporosis/terapia , Osteoporosis/prevención & control , Osteoporosis/sangre , Osteoporosis/metabolismo , Osteoporosis/patología , Masculino , Ratas , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Inflamación/terapia , Densidad Ósea/efectos de la radiación , Médula Ósea/efectos de la radiación , Médula Ósea/metabolismo , Microambiente Celular , EnvejecimientoRESUMEN
BACKGROUND AND AIMS: Chromosome evolution leads to hybrid dysfunction and recombination patterns and has thus been proposed as a major driver of diversification in all branches of the tree of life, including flowering plants. In this study we used the genus Linum (flax species) to evaluate the effects of chromosomal evolution on diversification rates and on traits that are important for sexual reproduction. Linum is a useful study group because it has considerable reproductive polymorphism (heterostyly) and chromosomal variation (nâ =â 6-36) and a complex pattern of biogeographical distribution. METHODS: We tested several traditional hypotheses of chromosomal evolution. We analysed changes in chromosome number across the phylogenetic tree (ChromEvol model) in combination with diversification rates (ChromoSSE model), biogeographical distribution, heterostyly and habit (ChromePlus model). KEY RESULTS: Chromosome number evolved across the Linum phylogeny from an estimated ancestral chromosome number of nâ =â 9. While there were few apparent incidences of cladogenesis through chromosome evolution, we inferred up to five chromosomal speciation events. Chromosome evolution was not related to heterostyly but did show significant relationships with habit and geographical range. Polyploidy was negatively correlated with perennial habit, as expected from the relative commonness of perennial woodiness and absence of perennial clonality in the genus. The colonization of new areas was linked to genome rearrangements (polyploidy and dysploidy), which could be associated with speciation events during the colonization process. CONCLUSIONS: Chromosome evolution is a key trait in some clades of the Linum phylogeny. Chromosome evolution directly impacts speciation and indirectly influences biogeographical processes and important plant traits.
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Lino , Linaceae , Filogenia , Lino/genética , Linaceae/genética , Fitomejoramiento , Poliploidía , Cromosomas , Evolución MolecularRESUMEN
Chemical mechanism (CM)-related surface-enhanced Raman spectroscopy (SERS) has received tremendous interest due to its exceptional stability and excellent uniformity. Nevertheless, there remains a demand for ingenious methodologies for promoting effective charge transfer (CT) to improve SERS sensitivity further. Herein, a band structure engineered W18O49/g-C3N4 heterostructure (WCN) was first employed as a CM-based SERS substrate with remarkable enhancement and sensitivity. To investigate the Raman enhancement properties of the substrate, malachite green (MG) was employed as the Raman probe with the excitation of a 633 nm laser. The WCN substrate exhibits a Raman enhancement factor (EF) of 2.6 × 107, achieving a limit of detection (LOD) of 1.9 × 10-10 M for MG. The outstanding Raman amplification behavior can be attributed to the heterojunction-induced efficient CT process, energy band matching resonance due to minor doping with g-C3N4 serving as a band gap modifier, and improved photo-induced charge transfer (PICT) efficiency via the oxygen vacancies in the W18O49 units. Additionally, a flexible SERS substrate based on WCN was constructed using a vacuum filtration method and utilized to detect prohibited pharmaceutical residues on fish skin. The integration of this WCN and a nylon membrane not only preserves the Raman activity of the WCN for sensitive detection but also endows the Raman substrate with high flexibility and good mechanical durability, making it a potential candidate for in situ detection in particular environments.
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IL-27, a heterodimeric cytokine of the IL-12 family, has diverse influences on the development of multiple inflammatory diseases. In this study, we identified the protective role of IL-27/IL-27R in host defense against Chlamydia muridarum respiratory infection and further investigated the immunological mechanism. Our results showed that IL-27 was involved in C. muridarum infection and that IL-27R knockout mice (WSX-1-/- mice) suffered more severe disease, with greater body weight loss, higher chlamydial loads, and more severe inflammatory reactions in the lungs than C57BL/6 wild-type mice. There were excessive IL-17-producing CD4+ T cells and many more neutrophils, neutrophil-related proteins, cytokines, and chemokines in the lungs of WSX-1-/- mice than in wild-type mice following C. muridarum infection. In addition, IL-17/IL-17A-blocking Ab treatment improved disease after C. muridarum infection in WSX-1-/- mice. Overall, we conclude that IL-27/IL-27R mediates protective immunity during chlamydial respiratory infection in mice by suppressing excessive Th17 responses and reducing neutrophil inflammation.
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Inflamación/inmunología , Interleucinas/inmunología , Neutrófilos/inmunología , Receptores de Interleucina/inmunología , Animales , Chlamydia muridarum/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores de Interleucina/deficiencia , Células Th17/inmunologíaRESUMEN
The IL-21/IL-21R interaction plays an important role in a variety of immune diseases; however, the roles and mechanisms in intracellular bacterial infection are not fully understood. In this study, we explored the effect of IL-21/IL-21R on chlamydial respiratory tract infection using a chlamydial respiratory infection model. The results showed that the mRNA expression of IL-21 and IL-21R was increased in Chlamydia muridarum-infected mice, which suggested that IL-21 and IL-21R were involved in host defense against C. muridarum lung infection. IL-21R-/- mice exhibited less body weight loss, a lower bacterial burden, and milder pathological changes in the lungs than wild-type (WT) mice during C. muridarum lung infection. The absolute number and activity of CD4+ T cells and the strength of Th1/Th17 responses in IL-21R-/- mice were significantly higher than those in WT mice after C. muridarum lung infection, but the Th2 response was weaker. Consistently, IL-21R-/- mice showed higher mRNA expression of Th1 transcription factors (T-bet/STAT4), IL-12p40, a Th17 transcription factor (STAT3), and IL-23. The mRNA expression of Th2 transcription factors (GATA3/STAT6), IL-4, IL-10, and TGF-ß in IL-21R-/- mice was significantly lower than that in WT mice. Furthermore, the administration of recombinant mouse IL-21 aggravated chlamydial lung infection in C57BL/6 mice and reduced Th1 and Th17 responses following C. muridarum lung infection. These findings demonstrate that IL-21/IL-21R may aggravate chlamydial lung infection by inhibiting Th1 and Th17 responses.
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Infecciones por Chlamydia/inmunología , Chlamydia muridarum/inmunología , Interleucinas/metabolismo , Pulmón/inmunología , Receptores de Interleucina-21/metabolismo , Subgrupos de Linfocitos T/inmunología , Células TH1/inmunología , Células Th17/inmunología , Animales , Femenino , Inflamación , Espacio Intracelular , Ratones , Receptores de Interleucina-21/genética , Factor de Transcripción STAT3/genética , Transducción de Señal , Proteínas de Dominio T Box/genéticaRESUMEN
BACKGROUND: Primary bilateral macronodular adrenal hyperplasia (PMAH) combined with infection by an opportunistic pathogen is complicated. Clinical evidence on managing PMAH patients with infections by opportunistic pathogens is insufficient. CASE PRESENTATION: A 66-year-old male was admitted with bilateral adrenal masses and was diagnosed with PMAH. Fever and disturbance of consciousness appeared after laparoscopic left adrenalectomy. Cryptococcal meningitis was confirmed by cerebrospinal fluid (CSF) culture. The exacerbation of his medical condition was suspected to result from immune reconstitution inflammatory syndrome (IRIS), and he had been treated with antifungal therapy and glucocorticoid replacement, but he responded poorly and eventually died of multiorgan failure. We summarized the clinical observations of 12 Cushing's syndrome (CS) patients infected by Cryptococcus. Seven out of nine patients who were treated for cryptococcus infection before receiving CS survived, while three patients treated for cryptococcus infection after CS treatment developed signs of IRIS and eventually died. CONCLUSION: Cushing's syndrome, complicated with cryptococcal infection, has a high mortality rate, mainly when IRIS emerges. Carefully identifying the presence of the suspected infection, and controlling cryptococcal infection before removing the culprit adrenals could be the rational choice.
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Criptococosis , Síndrome de Cushing , Meningitis Criptocócica , Masculino , Humanos , Anciano , Meningitis Criptocócica/diagnóstico , Meningitis Criptocócica/etiología , Adrenalectomía/efectos adversos , Síndrome de Cushing/etiología , Síndrome de Cushing/cirugía , Glándulas SuprarrenalesRESUMEN
OBJECTIVES: Tongue squamous cell carcinoma (TSCC) is the most common malignancy in oral cancer. Long noncoding RNAs (lncRNAs) are important regulators in cancer biology. In our present study, we investigated a novel lncRNA IGF-like family member 2 antisense RNA 1 (IGFL2-AS1) in TSCC. METHODS: RT-qPCR analyzed IGFL2-AS1 expression in TSCC cells. Functional assays assessed the impact of IGFL2-AS1 on TSCC cell proliferation, migration, and invasion. Western blot analyzed the protein levels of EMT-related markers. Mechanism assays analyzed the regulatory mechanism of IGFL2-AS1 in TSCC cells. In-vivo experiments were conducted to prove the role of IGFL2-AS1 in TSCC progression. RESULTS: IGFL2-AS1 was significantly up-regulated in TSCC cells and tissues, and IGFL2-AS1 knockdown inhibited cell proliferation, migration, invasion and EMT in TSCC. Moreover, IGFL2-AS1 functioned as a competing endogenous RNA (ceRNA) to sponge miR-1224-5p and thereby modulated SATB homeobox 1 (SATB1) expression. Additionally, SATB1 activated the Wnt/ß-catenin signaling pathway in TSCC cells and IGFL2-AS1 regulated the Wnt/ß-catenin signaling pathway and TSCC progression via elevating SATB1 expression. CONCLUSIONS: The data revealed that IGFL2-AS1 played a cancer promoting role in TSCC and may aid in exploring a brand new biomarker that might contribute to TSCC treatment.
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Carcinoma de Células Escamosas , Proteínas de Unión a la Región de Fijación a la Matriz , MicroARNs , ARN Largo no Codificante , Neoplasias de la Lengua , Humanos , Neoplasias de la Lengua/patología , Carcinoma de Células Escamosas/patología , Vía de Señalización Wnt/genética , Proteínas de Unión a la Región de Fijación a la Matriz/genética , Proteínas de Unión a la Región de Fijación a la Matriz/metabolismo , Línea Celular Tumoral , MicroARNs/genética , MicroARNs/metabolismo , Proliferación Celular/genética , Movimiento Celular/genética , Lengua , ARN Largo no Codificante/genética , Regulación Neoplásica de la Expresión GénicaRESUMEN
PURPOSE: To investigate the clinical characteristics and the risk factors associated with excessive daytime sleepiness (EDS) in patients with early- and late-onset narcolepsy. METHODS: Patients with narcolepsy were consecutively recruited. All patients were separated into early- and late-onset groups according to the onset age of disease ≤ 15 and > 15 years, respectively. Demographic, clinical, and sleep parameters were compared between the two groups. Linear regressions were performed to examine the risk factors of subjective and objective EDS in patients with early- and late-onset narcolepsy. RESULTS: A total of 101 patients with narcolepsy (median age at recruitment = 18.0 years) were classified into an early-onset group (67 patients with median age at onset = 12.0 years) and a late-onset group (34 patients with median age at onset = 28.5 years). Compared with early-onset group, late-onset group scored significantly higher on Epworth Sleepiness Scale (ESS), Ullanlinna Narcolepsy Scale (UNS), sleep paralysis, rapid eye movement (REM) sleep behavior disorder (RBD) questionnaire-Hong Kong (all P < 0.050). UNS-cataplexy and sleep paralysis had significantly positive associations with subjective EDS, and N1%, arousal index, and periodic limb movements index were positively associated with objective EDS in the early-onset group (all P < 0.050). However, these associations were not observed in late-onset narcolepsy. CONCLUSION: Late onset narcolepsy had more severe self-reported narcolepsy symptoms. REM sleep related symptoms and disrupted nighttime sleep were associated with EDS in early-onset narcolepsy. These findings suggest that early- and late-onset narcolepsy may represent two distinct phenotypes.
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Trastornos de Somnolencia Excesiva , Narcolepsia , Parálisis del Sueño , Humanos , Adulto , Adolescente , Polisomnografía , Narcolepsia/diagnóstico , Narcolepsia/epidemiología , Trastornos de Somnolencia Excesiva/diagnóstico , Trastornos de Somnolencia Excesiva/epidemiología , FenotipoRESUMEN
PURPOSE: To determine obstructive sleep apnea (OSA) phenotypes using cluster analysis including variables of sleep perception and sleep quality and to further explore factors correlated with poor sleep quality in different clusters. METHODS: This retrospective study included patients with OSA undergoing polysomnography (PSG) between December 2020 and April 2022. Two-step cluster analysis was performed to detect distinct clusters using sleep perception variables including discrepancy in total sleep time (TST), sleep onset latency (SOL), and wakefulness after sleep onset (WASO); objective TST, SOL, and WASO; and sleep quality. One-way analysis of variance or chi-squared tests were used to compare clinical and PSG characteristics between clusters. Binary logistic regression analyses were used to explore factors correlated with poor sleep quality. RESULTS: A total of 1118 patients were included (81.6% men) with mean age ± SD 43.3 ± 13.1 years, Epworth sleepiness score, 5.7 ± 4.4, and insomnia severity index 3.0 ± 2.4. Five distinct OSA clusters were identified: cluster 1 (n = 254), underestimated TST; cluster 2 (n = 158), overestimated TST; cluster 3 (n = 169), overestimated SOL; cluster 4 (n = 155), normal sleep discrepancy and poor sleep quality; and cluster 5 (n = 382), normal sleep discrepancy and good sleep quality. Patients in cluster 2 were older, more commonly had hypertension, and had the lowest apnea-hypopnea index and oxygen desaturation index. Age and sleep efficiency were correlated with poor sleep quality in clusters 1, 2, and 5, and also AHI in cluster 2. CONCLUSION: Subgroups of patients with OSA have different patterns of sleep perception and quality that may help us to further understand the characteristics of sleep perception in OSA and provide clues for personalized treatment.
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Apnea Obstructiva del Sueño , Trastornos del Inicio y del Mantenimiento del Sueño , Masculino , Humanos , Femenino , Calidad del Sueño , Estudios Retrospectivos , Sueño , Apnea Obstructiva del Sueño/diagnóstico , Apnea Obstructiva del Sueño/epidemiología , Análisis por Conglomerados , Fenotipo , PercepciónRESUMEN
OBJECTIVES: To explore associations between inflammatory endotypes and clinical presentations in CRS. To investigate the value of secretions myeloperoxidase (MPO) and eosinophilic cationic protein (ECP) detections in the diagnosis of endotypes of chronic rhinosinusitis (CRS), so as to provide guidance for the clinical application of MPO and ECP detection in secretions. METHODS: We collected clinical symptom scores from patients with CRS and examined the differences between endotypes in clinical features. Patients' nasal secretions and polyps (or middle turbinate for control) were collected and their NEU number, EOS%, MPO and ECP levels were measured. Correlation analysis was performed for these biomarkers in secretions and tissues, respectively. Receiver operating characteristic curves were used to assess the predictive potential of the biomarkers mentioned above in nasal secretions. RESULTS: Patients with Eos+Neu+ and Eos+Neu-CRS scored highest in most clinical symptom scores, while Eos-Neu+ and Eos-Neu-CRS scored lowest. Correlation analysis showed that tissues NEU number was correlated with NEU number and MPO level in nasal secretions (R = 0.4088; 0.6613); tissues EOS % was correlated with EOS% and ECP level in nasal secretions (R = 0.2344; 0.5774). To diagnose Neu+CRS, the highest area under the curve (AUC) (0.8961) was determined for MPO in secretions; the highest AUC (0.7400) was determined for NEU number in secretions. To diagnose Eos+Neu-CRS from Eos-Neu-CRS in Neu-CRS, the highest AUC (0.8801) was determined for ECP in secretions. CONCLUSIONS: Clinical presentations are directly associated with CRS endotypes. Measurement of MPO and ECP in nasal secretions is useful for the endotypes diagnosis of CRS.
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Pólipos Nasales , Rinitis , Sinusitis , Humanos , Rinitis/diagnóstico , Rinitis/metabolismo , Proteína Catiónica del Eosinófilo/metabolismo , Peroxidasa , Enfermedad Crónica , Sinusitis/diagnóstico , Sinusitis/metabolismo , Biomarcadores , Pólipos Nasales/diagnóstico , Pólipos Nasales/metabolismoRESUMEN
The incidence of Chlamydia trachomatis respiratory infection is increasing, and its pathogenesis is still unclear. Pyroptosis, as a mode of inflammatory cell death, plays a vital role in the occurrence and development of Chlamydia trachomatis respiratory infection. In this study, the potential pyroptosis-related genes involved in Chlamydia trachomatis respiratory infection were identified by constructing a mouse model of C. muridarum infection combined with bioinformatics analysis. Through in-depth analysis of the RNA sequencing data, 13 differentially expressed pyroptosis-related genes were screened, including 1 downregulated gene and 12 upregulated genes. Gene ontology (GO) analysis showed that these genes mainly regulate inflammatory responses and produce IL-1ß. Protein-protein interaction network analysis identified eight hub genes of interest: Tnf, Tlr2, Il1b, Nlrp3, Tlr9, Mefv, Zbp1 and Tnfaip3. Through quantitative real-time PCR (qPCR) analysis, we found that the expression of these genes in the lungs of C. muridarum-infected mice was significantly reduced, consistent with the bioinformatics results. At the same time, we detected elevated levels of caspase-3, gasdermin D and gasdermin E proteins in the lungs of C. muridarum-infected mice, demonstrating that Chlamydia trachomatis infection does induce pyroptosis. We then predicted nine miRNAs targeting these hub genes and constructed a key competitive endogenous RNA (ceRNA) network. In summary, we identified six key pyroptosis-related genes involved in Chlamydia trachomatis respiratory infection and constructed a ceRNA network associated with these genes. These findings will improve understanding of the molecular mechanisms underlying pyroptosis in Chlamydia trachomatis respiratory infections.
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Infecciones por Chlamydia , Chlamydia , MicroARNs , Animales , Ratones , Piroptosis/genética , Gasderminas , Infecciones por Chlamydia/genética , MicroARNs/genética , Proteínas de Unión al ARNRESUMEN
Interleukin-21 and its receptors (IL-21/IL-21R) aggravate chlamydial lung infection, while macrophages (Mφ) are one of the main cells infected by chlamydia and the main source of inflammatory cytokines. Therefore, it is particularly important to study whether IL-21/IL-21R aggravates chlamydia respiratory infection by regulating Mφ. Combined with bioinformatics analysis, we established an IL-21R-deficient (IL-21R-/-) mouse model of Chlamydia muridarum (C. muridarum) respiratory tract infection in vivo, studied C. muridarum-stimulated RAW264.7 by the addition of rmIL-21 in vitro, and conducted adoptive transfer experiments to clarify the association between IL-21/IL-21R and Mφ. IL-21R-/- mice showed lower infiltration of pulmonary total Mφ, alveolar macrophages, and interstitial macrophages compared with WT mice following infection. Transcriptomic analysis suggested that M1-related genes are downregulated in IL-21R-/- mice and that IL-21R deficiency affects the Mφ-mediated inflammatory response during C. muridarum infection. In vivo experiments verified that in IL-21R-/- mice, pulmonary M1-type CD80+, CD86+, MHC II+, TNFα+, and iNOS+ Mφ decreased, while there were no differences in M2-type CD206+, TGF-ß+, IL-10+ and ARG1+ Mφ. In vitro, administration of rmIL-21 to C. muridarum-stimulated RAW264.7 cells promoted the levels of iNOS-NO and the expression of IL-12p40 and TNFα, but had no effect on TGFß or IL-10. Further, adoptive transfer of M1-like bone marrow-derived macrophages derived from IL-21R-/- mice, unlike those from WT mice, effectively protected the recipients against C. muridarum infection and induced relieved pulmonary pathology. These findings help in understanding the mechanism by which IL-21/IL-21R exacerbates chlamydia respiratory infection by promoting the proinflammatory effect of Mφ.
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Infecciones por Chlamydia , Chlamydia muridarum , Animales , Ratones , Interleucina-10 , Factor de Necrosis Tumoral alfa , MacrófagosRESUMEN
The removal of antibiotic resistance genes (ARGs) in sewage is of great concern, but advanced sewage treatment technologies are not suitable for rural areas, so the multi-layer soil infiltration system (MSL) has been developed for rural sewage treatment. However, little is known about the performance and function of MSL in the treatment of ARGs in rural sewage. Here, we optimized the matrix composition and structure of MSL and explored the efficacy and mechanism of MSL systems for ARG removal under different hydraulic conditions. The ARGs removal rate of MSL ranged from 41.51% to 99.67%, in which MSL with the middle hydraulic load, high pollution load, and continuous inflowing conditions showed the best removal performance. In addition, this system can operate stably and resist the temperature fluctuation, which showed an equivalent removal rate of ARGs in warm and cold seasons, amounting to 69.0%. The structural equation model revealed that microorganisms in sewage could significantly affect ARG removal (path coefficient = 0.91), probably owing to their interspecies competition. As for the internal system, the reduction of ARGs was mainly driven by microorganisms in the system matrix (path coefficient = 0.685), especially soil-mixture-block (SMB) microorganisms. The physicochemical factors of the matrix indirectly reduce ARGs by affecting the microorganisms that adhere to the matrices. Note that the pairwise alignment of nucleotide analysis demonstrated that the system matrix, especially biochar in the SMB, adsorbed ARGs and their hosts from the sewage, and in turn eliminated them by inhibiting the spread and colonization of hosts, thereby reducing the abundance of ARGs. Collectively, this study provides a deeper insight into the removal of ARGs from rural sewage by MSL, which can help improve sewage treatment technologies.
RESUMEN
The emergence of antibiotic resistance is a severe threat to public health. There are few studies on the effects of sewage discharge on antibiotics and antibiotic resistance genes (ARGs) contamination in Jiaozhou Bay sediment. Herein, a total of 281 ARG subtypes, 10 mobile genetic elements (MGEs), 10 antibiotics and bacterial communities in marine sediments from Jiaozhou Bay were characterized. Similar bacterial community structures and ARG profiles were identified between the various sampling sites inside the bay, which were both dominated by multidrug and (fluoro)quinolone resistance genes and harbored lower relative abundances of ARGs than those in the sampling sites near the bay exit. Compared with antibiotics and MGEs, bacterial community composition was a more important driver of ARG diversity and geographic distribution. The abundance of pathogens carrying genetic information increased dramatically in southern Jiaozhou Bay is affected by sewage discharge, which indicating that wastewater discharge facilitated ARG contamination of marine sediments. This study highlights the risk of disseminating antibiotic resistance-influencing factors from treated wastewater discharge into marine environment there is an urgent need to optimize or improve wastewater treatment processes to enhance the removal of antibiotics and ARGs. This study has necessary implications for filling the gap in information on antibiotic resistance in Jiaozhou Bay and developing future pollution regulation and control measures.
Asunto(s)
Aguas del Alcantarillado , Aguas Residuales , Genes Bacterianos , Bahías , Bacterias/genética , Antibacterianos/farmacología , Farmacorresistencia Microbiana/genética , Sedimentos Geológicos , ChinaRESUMEN
Changes in stream biodiversity are now mainly driven by land-use development. However, a literature review on the impact of land use on stream macroinvertebrates is lacking, especially a scientometric review. Here, we bibliometrically analyzed the literature on land use and stream macroinvertebrates that were published in 2010-2021 and listed in the Web of Science database. We found that the impact of land use on stream macroinvertebrates had been increasingly studied and that these studies were distributed across the globe and had multi-national collaborations. Through co-citation analysis and high-frequency keyword analysis, we found that land use and some environmental factors, especially water quality and habitat, affected macroinvertebrate community biodiversity, biotic integrity, and patterns. Macroinvertebrate traits, analytical methods or models, evaluation index development, and riparian vegetation were the research hotspots. Using historical direct citation network analysis, we also found that the analytical methods in this field and the macroinvertebrate evaluation index had clear development trends from 2010 to 2021. Our findings can help researchers quickly grasp the background of the impact of land use on stream macroinvertebrates and inform future research.
Asunto(s)
Monitoreo del Ambiente , Invertebrados , Animales , Monitoreo del Ambiente/métodos , Ríos , Ecosistema , BibliometríaRESUMEN
Chinese government is vigorously promoting toilet renovation in rural areas to reduce the risk of human feces exposure, which would cause infectious diseases, especially antibiotic resistance genes (ARGs) and pathogens. However, the distribution of ARGs in human feces from different regions of China remained ill-defined. It is not yet known how the survival of ARGs after toilet treatment is associated with the regional infection rates. Here, we investigated the prevalence of ARGs in human feces in rural areas of China and their potential relationship with infectious diseases for the first large-scale. The results showed that there were still high ARGs residues in human feces after rural toilet treatment, especially tetM-01 and ermB with average relative abundance as high as 1.21 × 10-1 (Eastern) and 1.56 × 10-1 (Northern), respectively. At a large regional scale, the significant differences in human feces resistomes were mainly shaped by the toilet types, TN, NH3-N, and the bacterial community. A critical finding was that toilets still cannot effectively decrease the pathogenicity risk in human feces. The significant positive relationship (P<0.05) between infectious diseases and ARGs can infer that ARGs in human feces exposure might be a critical path for enhancing the incidence of diseases, as these ARGs hinder the effectiveness of antibiotics.