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BiVO4 is a promising photoanode for photoelectrochemical (PEC) water splitting but suffers from high charge carrier recombination and sluggish surface water oxidation kinetics that limit its efficiency. In this work, a model of sulfur-incorporated FeOOH cocatalyst-loaded BiVO4 was constructed. The composite photoanode (BiVO4/S-FeOOH) demonstrates an enhanced photocurrent density of 3.58 mA cm-2, which is 3.7 times higher than that of the pristine BiVO4 photoanode. However, the current explanations for the generation of enhanced photocurrent signals through the incorporation of elements and cocatalyst loading remain unclear and require further in-depth research. In this work, the hole transfer kinetics were investigated by using a scanning photoelectrochemical microscope (SPECM). The results suggest that the incorporation of sulfur can effectively improve the charge transfer capacity of FeOOH. Moreover, the oxygen evolution reaction model provides evidence that S-doping can induce a "fast" surface catalytic reaction at the cocatalyst/solution interface. The work not only presents a promising approach for designing a highly efficient photoanode but also offers valuable insights into the role of element doping in the PEC water-splitting system.
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Limited charge separation/transport efficiency remains the primary obstacle of achieving satisfying photoelectrochemical (PEC) water splitting performance. Therefore, it is essential to develop diverse interfacial engineering strategies to mitigate charge recombination. Despite obvious progress having been made, most works only considered a single-side modulation in either the electrons of conduction band or the holes of valence band in a semiconductor photoanode, leading to a limited PEC performance enhancement. Beyond this conventional thinking, we developed a novel coupling modification strategy to achieve a composite electrode with bidirectional carrier transport for a better charge separation, in which Ti2 C3 Tx MXene quantum dots (MQDs) and α-Fe2 O3 nanodots (FO) are anchored on the surface of ZnIn2 S4 (ZIS) nanoplates, resulting in markedly improved PEC water splitting of pure ZIS photoanode. Systematic studies indicated that the bidirectional charge transfer pathways were stimulated due to MQDs as "electron extractor" and S-O bonds as carriers transport channels, which synergistically favors significantly enhanced charge separation. The enhanced kinetic behavior at the FO/MQDs/ZIS interfaces was systematically and quantitatively evaluated by a series of methods, especially scanning photoelectrochemical microscopy. This work may deepen our understanding of interfacial charge separation, and provide valuable guidance for the rational design and fabrication of high-performance composite electrodes.
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Photoelectrochemical (PEC) water splitting technology is a promising strategy toward producing sustainable hydrogen fuel. However, it is an essential bottleneck to reduce severe charge recombination for the improvement of PEC performance. Construction of heterojunction systems, such as Z-scheme and type II heterojunctions, could efficiently boost charge separation, whereas the mechanism of charge separation is still ambiguous. We describe herein a charge transfer system designed with Bi2WO6/Bi2S3 (BWO/BS) as a prototype. In this system, Au nanoparticles act as charge relays to engineer a charge transfer pathway, and the obtained BWO/Au/BS photoanode achieves a remarkable photocurrent density of 0.094 mA cm-2 at 1.23 V versus reversible hydrogen electrode (vs RHE), over approximately 1.2 and 2.3 times larger than those of BWO/BS/Au and BWO, exhibiting long-term photostability. More importantly, scanning photoelectrochemical microscopy (SPECM) and intensity-modulated photocurrent spectroscopy (IMPS) studies are performed to in situ-capture the photogenerated hole during the PEC process. Operando analysis reveals that the Z-scheme BWO/Au/BS system (1.33 × 10-2 cm s-1) exhibits higher charge transfer kinetics compared to the type II BWO/BS/Au heterostructure (0.85 × 10-2 cm s-1) while efficiently suppressing charge recombination for optimized PEC activity. Note that this smart strategy can also be extended to other semiconductor-based photoanodes such as BiVO4. Our study offers an effective pathway for the rational design of highly efficient charge separation for solar conversion based on water splitting.
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Semiconductor/co-catalyst coupling is considered as a promising strategy to enhance the photoelectrochemical (PEC) conversion efficiency. Unfortunately, this model system is faced with a serious interface recombination problem, which limits the further improvement of PEC performances. Here, a FeNiOOH co-catalyst with abundant oxygen vacancies on BiVO4 is fabricated through simple and economical NaBH4 reduction to accelerate hole transfer and achieve efficient electron-hole pair separation. The photocurrent of the BV (BiVO4 )/Vo-FeNiOOH system is more than four times that of pure BV. Importantly, the charge transfer kinetics and charge carrier recombination process are studied by scanning photoelectrochemical microscopy and intensity modulated photocurrent spectroscopy in detail. In addition, the oxygen vacancy regulation proposed is also applied successfully to other semiconductors (Fe2 O3 ), demonstrating the applicability of this strategy.
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Oxigênio , Semicondutores , Catálise , Oxigênio/químicaRESUMO
OBJECTIVE: To investigate the correlation of circulating long non-coding RNA nuclear-enriched abundant transcript 1 (lncRNA NEAT1) expression with disease risk, severity, prognosis and inflammatory cytokine levels in sepsis patients. METHODS: 152 sepsis patients and 150 health controls (HCs) were enrolled in this study. Plasma and serum samples were obtained from sepsis patients and HCs, and lncRNA NEAT1 expression in plasma was determined by quantitative polymerase chain reaction, while levels of inflammatory cytokines in serum were detected by enzyme linked immune sorbent assay. RESULTS: LncRNA NEAT1 expression was remarkably higher in sepsis patients than in HCs (Pâ¯<â¯0.001). Receiver operating characteristic (ROC) curve disclosed a good predictive value of lncRNA NEAT1 expression for sepsis risk with area under curve (AUC) of 0.730 (95% CI: 0.740-0.861). Subsequent multivariate logistic regression analysis demonstrated that lncRNA NEAT1 expression was independently associated with higher sepsis risk (Pâ¯<â¯0.001). In sepsis patients, lncRNA NEAT1 expression was also observed to be positively correlated with Acute Physiology and Chronic Health Evaluation (APACHE) II score (Pâ¯<â¯0.001), serum tumor necrosis factor-α (Pâ¯<â¯0.001), interleukin (IL)-1ß (Pâ¯=â¯0.043), IL-6 (Pâ¯=â¯0.001) and IL-8 (Pâ¯=â¯0.038), while negatively correlated with IL-10 (Pâ¯<â¯0.001). In addition, lncRNA NEAT1 expression was increased in non-survivors compared to survivors (Pâ¯=â¯0.006), and ROC curve revealed a good prognostic value of lncRNA NEAT1 for non-survivor risk with AUC 0.641 (95% CI: 0.536-0.746). CONCLUSION: Circulating lncRNA NEAT1 correlates with increased disease risk, elevated severity and unfavorable prognosis as well as higher expression of pro-inflammatory cytokines in sepsis patients.
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RNA Longo não Codificante/sangue , Sepse/diagnóstico , Biomarcadores/sangue , Estudos de Casos e Controles , Citocinas/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Sepse/sangue , Índice de Gravidade de DoençaRESUMO
Background: The anti-inflammatory effects of budesonide (BUN) and N-acetylcysteine (NAC) attenuate acute lung injury (ALI). The aim of this study was to investigate the effects of combination therapy consisting of BUN and NAC on ALI and the underlying mechanisms. Methods: In vitro and in vivo models of ALI were generated by LPS induction. Western blotting was used to detect the expression levels of pyroptosis-related proteins and inflammation-related factors, and RT-qPCR was used to detect the expression of miR-381. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. ELISA was used to detect the levels of inflammation-related factors. HE staining was used to detect lung injury. Results: The results showed that LPS effectively induced pyroptosis in cells and promoted the expression of pyroptosis-related proteins (Caspase1, Gasdermin D and NLRP3) and inflammatory cytokines (TNF-α, IL-6 and IL-1ß). The combination of BUN and NAC significantly alleviated LPS-induced pyroptosis and inflammation. In addition, the combination of BUN and NAC effectively promoted miR-381 expression. Transfection of miR-381 mimics effectively alleviated LPS-induced pyroptosis and inflammation, while transfection of miR-381 inhibitors had the opposite effect. miR-381 negatively regulates NLRP3 expression. Treatment with a miR-381 inhibitor or pc-NLRP3 reversed the effects of the combination of BUN and NAC. In a mouse model of ALI, the combination of BUN and NAC effectively improved lung injury, while treatment with a miR-381 inhibitor or pc-NLRP3 effectively reversed this effect. Conclusion: Overall, this study revealed that BUN + NAC inhibits the activation of NLRP3 by regulating miR-381, thereby alleviating ALI caused by pyroptosis-mediated inflammation.
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Human metapneumovirus (hMPV) causes acute respiratory infections in children. The prevalence and clinical characteristics of hMPV were determined in nasopharyngeal aspirates of children in Changsha, China. Reverse transcription-polymerase chain reaction (RT-PCR) or PCR was employed to screen for both hMPV and other common respiratory viruses in 1,165 nasopharyngeal aspirate specimens collected from children with lower respiratory tract infections from September 2007 to August 2008. All PCR products were sequenced, and demographic and clinical data were collected from all patients. Seventy-six of 1,165 (6.5%) specimens were positive for hMPV, of which 85.5% (65/76) occurred in the winter and spring seasons. The hMPV coinfection rate was 57.9% (44/76), and human bocavirus was the most common virus detected in conjunction with hMPV. Phylogenetic analysis revealed that 94.7% of the hMPV detected were of subgroup A2, 5.3% were subgroup B2, and none belonged to either the A1 or B1 subgroups. No significant differences were found in terms of the frequency of diagnosis and clinical signs between either the co- and mono-infection groups, or between patients with and without underlying diseases. It was concluded that hMPV is an important viral pathogen in pediatric patients with lower respiratory tract infections in Changsha. Only hMPV genotypes A2 and B2 were co-circulating in this locality; human bocavirus was the most common coinfecting virus, and coinfection did not affect disease severity.
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Metapneumovirus/isolamento & purificação , Infecções por Paramyxoviridae/epidemiologia , Infecções por Paramyxoviridae/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Adolescente , Criança , Pré-Escolar , China/epidemiologia , Coinfecção/epidemiologia , Coinfecção/virologia , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Epidemiologia Molecular , Nasofaringe/virologia , Filogenia , Prevalência , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
BACKGROUND: Human adenovirus (HAdV) is an important agent causing respiratory tract infection in children. Information on the epidemiological and clinical features of HAdV is limited in children with acute respiratory tract infections (ARTIs) in China, especially those of a novel genotype, Ad55. METHODS: In total, 1169 nasopharyngeal aspirates were collected from children younger than 14 years with ARTIs between November 2006 and November 2009. The polymerase chain reaction (PCR) was used to screen HAdVs. All PCR-positive products were sequenced. RESULTS: 74 of 1169 (6.33%) specimens were positive for HAdVs. Among positive cases, AdV3 (58/74) was detected most frequently, followed by AdV11 (10/74), AdV2 (2/74), AdV7 (2/69), AdV6 (1/74), and AdV1 (1/74). AdV55 was found in one case. The incidence of HAdV infection peaked in children aged 3-7 years. The most common clinical diagnosis was upper respiratory infection, and the most common syndrome was fever and cough.The comparison of HAdV and RSV group revealed that Children infected with group AdV were significant older than children infected with group RSV, had more fever but less frequently wheezing, and cough, crackles, and cyanosis, The duration of hospitalization between the AdV group and RSV group was not significant, but a greater frequency of LRTIs was observed in RSV group. CONCLUSIONS: HAdV is an important viral agent in children with ARTIs in Lanzhou City, China. Multiple HAdV serotypes co-circulated with Ad3, which was predominant in this 3-year study. The novel AdV55 genotype was found in one case. No fixed seasonal rhythm could be identified.
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Infecções por Adenoviridae/epidemiologia , Adenovírus Humanos/isolamento & purificação , Infecções Respiratórias/epidemiologia , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Adolescente , Criança , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Nasofaringe/virologia , Reação em Cadeia da Polimerase , Prevalência , SorotipagemRESUMO
This study assessed the effects of a 12-week afterschool mindfulness-based diet and exercise intervention on mental and physical health in Latinx and Black youth. One hundred forty-eight boys and girls (average age = 10.1 years, SD = 1.3 years; 52% girls; 72.3% Latinx) were randomized to either the experimental group (n = 80) or the control group (n = 68). The experimental group participants engaged in fitness yoga, kickboxing, and/or spinning sessions, and mindfulness practices (e.g., breathing, meditation, and mindful eating) twice per week for 12 weeks. The control group participants engaged in a recreational play session once per week for 12 weeks. All participants completed surveys (demographics, acculturation, anxiety, emotional eating, sleep, and food intake) and had their height, weight, and percent body fat measured pre- and post-intervention. Participants wore an accelerometer for 7 days pre- and post-intervention. Repeated measures analysis of covariance indicated that the experimental group participants reported lower scores in emotional eating, anxiety, and sleep latency post-intervention compared to the control group participants. Conversely, no significant differences were observed in physical activity between the experimental and control group participants post-intervention. These findings indicate that a mindfulness-based intervention has a positive effect on emotional eating, anxiety, and sleep latency among youth of color.
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Bismuth vanadate (BiVO4) has been considered a promising photoactive material in photoelectrochemical (PEC) water-splitting systems. However, the performance of BiVO4-based photoanodes is currently unsatisfactory, indicating the need for new architectural designs to improve their efficiency. In this paper, a porphyrin-phosphazene polymer (THPP-HCCP) was synthesized with a sizeable conjugated structure, and Ag particles were deposited on its surface as an organic-inorganic composite interface improvement layer. The deposition of the composite polymer film on BiVO4 resulted in a significant increase in photocurrent density, reaching up to 2.2 mA cm-2 (1.23 V vs. RHE), almost three times higher than pristine BiVO4, which benefits from the synergistic effect of Ag nanoparticles and porphyrin-phosphazene. Furthermore, photophysical and intensity-modulated photocurrent analysis demonstrated that the Ag-THPP-HCCP heterostructures could broaden the light-absorbing range and facilitate hole transfer to the semiconductor surface, resulting in an improved water oxidation process. The dynamic charge transport behavior of Ag-THPP-HCCP/BiVO4 was investigated using scanning photoelectrochemical microscopy, which showed that the rate constant (Keff) exhibits an almost 4-fold increase compared to pristine BiVO4, indicating a significant improvement in the transport of photogenerated holes. This experiment presents a novel strategy for designing high-efficiency polymer-based photoanodes.
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BACKGROUND: Allergic rhinitis (AR) is characterized by distinct clinical heterogeneity and allergic sensitization patterns. We aimed to quantify rhinitis symptoms in patients with self-reported allergic rhinitis according to the potential sensitization patterns for relevant allergens in China. METHODS: We used latent class analysis (LCA; a subset of structural equation modeling) to independently cluster patients into different patterns of atopic sensitization in an unsupervised manner, based on specific immunoglobulin E tests. AR symptom severity was assessed by the visual analogue scale. We evaluated the association between the severity of AR and the allergen sensitization patterns. RESULTS: LCA revealed four phenotypes of atopic sensitization among 967 patients with self-report AR. We labeled latent classes as: Class 1, weed pollens and indoor sensitization (n = 74 [7.7%]); Class 2, weed pollen with low indoor sensitization (n = 275 [28.4%]); Class 3, low or no sensitization (n = 350 [36.2%]); and Class 4, house dust mite-dominated sensitization (n = 268 [27.7%]). AR was more severe in Class 2 compared to the other 3 classes, indicating that upper respiratory symptoms are more severe among patients with isolated seasonal rhinitis. CONCLUSION: We have identified four sensitization patterns in patients with self-reported AR, which were associated with different clinical symptoms and comorbidities.
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Rinite Alérgica , Rinite , Animais , Autorrelato , Estudos Transversais , Rinite Alérgica/diagnóstico , Rinite Alérgica/epidemiologia , Alérgenos , China/epidemiologia , Testes CutâneosRESUMO
ADAR (adenosine deaminase acting on RNA) is an RNA editing enzyme that targets both coding and noncoding dsRNAs (double stranded RNAs) and converts adenosine to inosine, which is read by translation machinery and by polymerases during RNA-dependent RNA replication as if it is guanosine. This editing is a widespread post-transcriptional modification event in animals. In this study, we identified the full-length cDNA sequence of Ctenopharyngodon idella ADAR1 (designated as CiADAR1) and detected the mRNA expression patterns in response to dsRNA (polyinosinic-polycytidylic acid sodium salt, poly(I:C)) and grass carp reovirus (GCRV). CiADAR1 is a large gene in size, consisting of 4833 nucleotides encoding a protein of 1392 amino acids. The deduced amino acid sequence contains seven putative domains: one proline-rich region (Pro-R), two Z-DNA-binding domains (Zalpha), three dsRNA binding motifs (DSRM) and one tRNA-specific and dsRNA adenosine deaminase domain (ADEAMc). It is most homologous to Danio rerio ADAR (E-value = 0.0, identities = 80% (1110/1395)), also close homology to Homo sapiens ADAR1 (E-value = 0.0, identities = (47%) 530/1122). CiADAR1 mRNA was investigated in fifteen tissues, and was low expressed in muscle and head kidney tissues, high in blood and spleen tissues by quantitative real-time RT-PCR (qRT-PCR). mRNA expressions of CiADAR1 were significantly up-regulated and reached peak at 24 h post GCRV challenge in vivo and in vitro (P < 0.05). After poly(I:C) stimulation at different concentrations, CiADAR1 transcripts were rapidly and significantly up-regulated and recovered in dose-dependent and time-dependent manners (P < 0.05). The results indicate CiADAR1 was implicated in the antiviral immune response and laid the foundation for further studies on functions and mechanisms of RNA editing in fishes.
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Adenosina Desaminase/genética , Adenosina Desaminase/imunologia , Carpas/genética , Carpas/imunologia , Regulação da Expressão Gênica , Adenosina Desaminase/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Carpas/metabolismo , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Perfilação da Expressão Gênica/veterinária , Dados de Sequência Molecular , Especificidade de Órgãos , Filogenia , Edição de RNA , RNA de Cadeia Dupla/metabolismo , Proteínas de Ligação a RNA , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reoviridae , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência/veterináriaRESUMO
OBJECTIVE: the aim of this retrospective study was to explore the clinical risk factors correlated to the prognosis of patients who suffered from central nervous system infection after neurosurgery. METHODS: the study included 113 patients who were diagnosed with postoperative intracranial infection. Several factors with clinical relevance were identified and analyzed by univariate analyses. The risk factors that showed any significant difference between the cases were analyzed by multivariate logistic regression analyses. RESULTS: Here we show that the duration of the drainage before infection (measured in days) (B: -0.113, OR: 0.893, 95% CI: 0.805-0.991, P: 0.033), the number of antibiotics used for the treatment (B:-1.470, OR: 0.230, 95% CI: 0.072-0.738, P: 0.013), the number of leucocytes in the cerebrospinal fluid (CSF) (B: -0.016, OR: 0.984, 95% CI: 0.970-0.998, P: 0.027) are risk factors for the prognosis of patients with intracranial infection. Instead, the duration of antibiotic treatment (measured in days) (B: 0.176, OR: 1.193, 95% CI: 1.063-1.339, P: 0.003) resulted to be a positive factor for the recovery from the infection. CONCLUSIONS: our results suggest that the prognosis of patients with intracranial infection after neurosurgery can be improved by an early identification of these correlated risk factors.
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BACKGROUND: Paediatric pleural tuberculosis (TB) is a paucibacillary disease, which increases the difficulty of examination. We aimed to assess the performance of pleural fluid adenosine deaminase (ADA) in the detection of paediatric pleural TB. METHODS: PubMed, Web of Science Core Collection, Embase and Cochrane Library databases were searched up to 20 December 2021. We used the bivariate and hierarchical summary receiver operating characteristic models to compute pooled estimates for the overall diagnostic accuracy parameters of ADA for diagnosing paediatric pleural TB. RESULTS: Eight studies, including 290 pleural fluid samples, met the inclusion criteria. The pooled sensitivity of ADA was 0.85 (95% CI: 0.78-0.90, I2: 55.63% < 75%) for detecting patients with paediatric pleural TB. A total of 262 pleural fluid samples from four studies were included to differentiate patients with paediatric pleural TB from controls. At a unified cut-off value of 40 U/L, the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio and area under the summary receiver operating characteristic curve of ADA were 0.89, 0.58, 2.09, 0.20, 10.48 and 0.89, respectively. CONCLUSIONS: At a cut-off value of 40 U/L, the overall performance of ADA was good for detecting paediatric pleural TB, with relatively high sensitivity and low specificity. Key messageAccurate identification of paediatric pleural TB will help eliminate TB in children. At a cut-off value of 40 U/L, the overall performance of ADA was good for detecting paediatric pleural TB, with relatively high sensitivity and low specificity.
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Derrame Pleural , Tuberculose Pleural , Humanos , Criança , Tuberculose Pleural/diagnóstico , Adenosina Desaminase , Derrame Pleural/diagnóstico , Sensibilidade e Especificidade , Curva ROCRESUMO
Photoelectrochemical (PEC) water splitting is an attractive strategy to convert and store of intermittent solar power into fuel energy. However, the detrimental charge recombination of photogenerated electrons and holes severely limits its efficiency. Despite electrocatalyst loading can obviously improve the PEC conversion efficiency, current systems still suffer from high recombination owing to the surface states. Herein, an interface "repairing" strategy is proposed to suppress the recombination at the semiconductor/electrocatalyst interface. NiOx layer acts as an interfacial repairing layer to efficiently extract photogenerated charge carriers and eliminate the surface states via high hole-transfer kinetics rather than as a traditional electrocatalyst. As expected, the resulting repaired system yields an impressive photocurrent density of 4.58 mA cm-2 at 1.23 V (vs. RHE), corresponding to a more than three-fold increase compared to BiVO4 (1.40 mA cm-2). Our work offers an appealing maneuver to improve the water oxidation performance for the semiconductor/electrocatalyst coupling system.
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Background: Because of the significant regional differences in the distribution of allergens, the relationship between anaphylaxis and allergic sensitization is complex in China. Using this large-scale epidemiologic survey, we explore the potential patterns of sensitization to common allergens in mainland China and investigate their relationship with various clinical symptoms. Method: The participants were recruited from 13 medical centers in mainland China from October 2019 to June 2021. Skin prick test (SPT) results that cover 18 common allergens were utilized to diagnose atopic sensitization. The demographic characteristics and clinical information were collected through questionnaires during routine medical follow-up. Latent class analysis (LCA) was conducted to determine the optimal sensitization patterns. The logistic regression was used to assess the associations of different sensitization patterns with allergy symptoms. Findings: A total of 1089 patients who had a positive SPT to at least one of 18 allergens were included for formal analysis. An optimal LCA model with 4 classes was obtained in this study, and the corresponding labels were as follows: Class1, house dust mite sensitization; Class2, low pollen sensitization; Class3, middle pollen sensitization; Class4, high pollen sensitization. The prevalence of different classes varied widely in geographical distribution, which was characterized by Class1 being very common in south and east as well as Class2 in north and west of China. Compared with patients in Class1, those in middle and high pollen sensitization clusters had the higher odds ratios (ORs) of allergic rhinitis and allergic conjunctivitis when controlling for other confounders. However, there was no significant difference between low pollen sensitization and house dust mite sensitization groups in the risks for various clinical performances except dermatitis. Additionally, the adjusted ORs (95% confidence interval) of allergic conjunctivitis and dermatitis for participants in pollen sensitization clusters (Class2, 3 and 4) were 1.56 (1.18, 2.06) and 1.43 (1.09, 1.88) respectively compared with those in Class1. Interpretation: In this study, we identified four sensitization clusters with specific risks of various clinical symptoms using common allergens by adopting LCA. Our findings may contribute to improved diagnosis and potential immunotherapy approaches to allergy in mainland China. Funding: This study was supported by the National Natural Science Foundation of China (81802076 and 81871736), the Guangzhou Science and Technology Foundation (202102010327), the Foundation of SKLRD (MS-2019-06 and Z-2022-09), and the Foundation of GYYY (ZH201904) and ZNSA-2020012.
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Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) has become one of the most commonly used techniques for RNA expression. To obtain more reliable results with biological significance, it requires data normalization using an appropriate internal control gene. Here, we cloned partial sequence of elongation factor 1α (EF1α) gene from grass carp (Ctenopharyngodon idella). The stabilities of four commonly used internal control genes encoding 18S rRNA, ß-actin, EF1α, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were integratedly assessed using the geNorm, NormFinder and BestKeeper programs. Integrative analyses of qRT-PCR data indicated that the stability ranking of the examined genes was 18S rRNA > EF1α > GAPDH > ß-actin in gill, head kidney, heart, intestine, liver, muscle, skin, spleen, and trunk kidney tissues in untreated grass carp. When the same calculations were done in spleen tissue at different time points post grass carp reovirus (GCRV) infection, the gene ranking was 18S rRNA > ß-actin > GAPDH > EF1α. The rank ordering of expression stability was EF1α > ß-actin>18S rRNA > GAPDH in C. idella kidney (CIK) cell culture stimulated by poly(I:C). The recommended ranking was EF1α > GAPDH > ß-actin>18S rRNA in CIK cells infected by GCRV. The results indicated that 18S rRNA was the best invariant internal control gene in individual level in grass carp, EF1α was the most suitable in CIK cell culture stimulated by poly(I:C) or infected by GCRV. As an assay, EF1α was employed to examine the changes of Toll-like receptor 3 (TLR3) and melanoma differentiation associated gene 5 (MDA5) after virus infection in CIK cells. These data laid the foundation for more precise results in qRT-PCR studies of gene expression in grass carp.
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Carpas , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Perfilação da Expressão Gênica/veterinária , Regulação da Expressão Gênica/efeitos dos fármacos , Infecções por Reoviridae/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adjuvantes Imunológicos/farmacologia , Animais , Carpas/genética , Carpas/metabolismo , Linhagem Celular , RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/metabolismo , Perfilação da Expressão Gênica/métodos , Poli I-C/farmacologia , Reoviridae/fisiologia , Infecções por Reoviridae/imunologia , Baço/metabolismo , Fatores de Tempo , Receptor 3 Toll-Like/genética , Receptor 3 Toll-Like/metabolismoRESUMO
IPS-1 (interferon-ß promoter stimulator 1), also known as MAVS/VISA/Cardif, plays a central role in antiviral immunity. In this manuscript, we cloned and characterized IPS-1 from grass carp Ctenopharyngodon idella (designated as CiIPS-1). The CiIPS-1 cDNA is 2412 bp long and consists of a 5' untranslated region (UTR) of 124 bp, a 3' UTR of 497 bp with three cytokine RNA instability motifs (ATTTA) and a polyadenylation signal (AATAAA), and an open reading frame (ORF) of 1791 bp encoding a polypeptide of 596 amino acids with a calculated molecular mass of 64.1 kDa and a theoretical isoelectric point of 4.79. Structural analysis showed that the CiIPS-1 protein contained an N-terminal CARD (caspase activation and recruitment domain), a central proline-rich domain, a putative TRAF2-binding motif and a C-terminal transmembrane domain. Similarity analysis of the deduced amino acid sequence of the CiIPS-1 by MatGAT software revealed that the CiIPS-1 shared 27.8-76.4% identity and 47.4-85.2% similarity with other known piscine IPS-1 sequences. The CiIPS-1 mRNA was constitutively expressed in the examined tissues, higher in spleen, and was induced by grass carp reovirus (GCRV) injection by semi-quantitative RT-PCR assay. Quantitative real-time RT-PCR analysis revealed that the CiIPS-1 mRNA expression was rapidly and significantly up-regulated in vivo and in vitro after GCRV infection, and the CiIPS-1 transcripts were also significantly enhanced in vitro post the synthetic double stranded RNA polyinosinic-polycytidylic potassium salt (poly(I:C)) stimulation. These results indicated that CiIPS-1 was an inducible acute-phase protein and involved in the immune reaction to GCRV in grass carp.
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Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Carpas , Clonagem Molecular , Regulação da Expressão Gênica/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/genética , Dados de Sequência Molecular , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reoviridae , Infecções por Reoviridae/metabolismo , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologiaRESUMO
RIG-I (retinoic acid inducible gene-I) is a key mediator of antiviral immunity, able to couple detection of infection by RNA and DNA viruses to the induction of interferons. In the present study, a RIG-I gene from grass carp Ctenopharyngodon idella (CiRIG-I) was isolated and characterized. The full-length cDNA of CiRIG-I was of 3198 bp and encoded a polypeptide of 947 amino acids with an estimated molecular mass of 108,730 Da and a predicted isoelectric point of 5.85, including six main overlapping structural domains: two CARDs (caspase activation and recruitment domain), one ResIII (conserved restriction domain of bacterial type III restriction enzyme), one DEXDc (DEAD/DEAH box helicase domain), one HELICc (helicase superfamily c-terminal domain) and one RD (regulatory domain). The CiRIG-I mRNA was widespread expression in the tested 15 tissues by semi-quantitative RT-PCR (sqRT-PCR) assay. The CiRIG-I expressions in spleen and liver were significantly induced following grass carp reovirus (GCRV) infection. CiRIG-I mRNA expression was rapidly and significantly up-regulated in vitro after GCRV infection, and the CiRIG-I transcripts were also significantly enhanced in vitro post the synthetic double stranded RNA polyinosinic-polycytidylic potassium salt (poly(I:C)) stimulation. These results collectively suggested that CiRIG-I was an inducible protein, involved in the antiviral innate immune defense to GCRV in grass carp, and laid the foundation for the further mechanism research of RIG-I in fishes.
Assuntos
Carpas/genética , Carpas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Adjuvantes Imunológicos/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Carpas/classificação , Doenças dos Peixes/imunologia , Perfilação da Expressão Gênica , Imunidade Inata , Fígado/efeitos dos fármacos , Fígado/imunologia , Dados de Sequência Molecular , Filogenia , Poli I-C/farmacologia , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Baço/efeitos dos fármacos , Baço/imunologia , Fatores de Tempo , Fatores de Transcrição/químicaRESUMO
Human asthma is caused by interactions between a range of genetic and environmental factors. However, the specific pathogenesis of asthma remains controversial. This study explored the contribution of DNA methylation to asthma using computer learning methods. Relevant datasets and information related to patients with asthma were collected from the Gene Expression Omnibus (GEO) database. A multivariate linear regression model was established. Differentially expressed genes and DNA methylation sites were identified. The results showed that the expression of 169 genes was significantly different between the two groups. Through differential analysis of methylation and differential analysis of gene expression, 44 differentially expressed genes that may be affected by DNA methylation modification were identified. The results of the multiple linear regression model showed that DNA methylation could explain 9.81% of the variation in gene expression. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that the differentially expressed genes, HLA-DMB, IL4, HLA-DPB1, and CD40LG, were related to the occurrence of asthma, and HLA-DMB expression was significantly reduced in allergic asthma. There was a positive correlation between cg04933135 and HLA-DMB expression, and cg04933135 was a differential site for DNA methylation. Using blood samples from asthma patients, we confirmed that HLA-DMB expression is down-regulated, which may be affected by abnormal DNA methylation. DNA methylation plays an important role in the development of asthma, and HLA-DMB which modified by abnormal DNA methylation can be regarded as a new biomarker of asthma.