RESUMO
The increased soil salinity is becoming a major challenge to produce more crops and feed the growing population of the world. In this study, we demonstrated that overexpression of OsDIR55 gene enhances rice salt tolerance by altering the root diffusion barrier. OsDIR55 is broadly expressed in all examined tissues and organs with the maximum expression levels at lignified regions in rice roots. Salt stress upregulates the expression of OsDIR55 gene in an abscisic acid (ABA)-dependent manner. Loss-function and overexpression of OsDIR55 compromised and improved the development of CS and root diffusion barrier, manifested with the decreased and increased width of CS, respectively, and ultimately affected the permeability of the apoplastic diffusion barrier in roots. OsDIR55 deficiency resulted in Na+ accumulation, ionic imbalance, and growth arrest, whereas overexpression of OsDIR55 enhances salinity tolerance and provides an overall benefit to plant growth and yield potential. Collectively, we propose that OsDIR55 is crucial for ions balance control and salt stress tolerance through regulating lignification-mediated root barrier modifications in rice.
Assuntos
Regulação da Expressão Gênica de Plantas , Oryza , Proteínas de Plantas , Raízes de Plantas , Tolerância ao Sal , Oryza/genética , Oryza/fisiologia , Oryza/metabolismo , Oryza/crescimento & desenvolvimento , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Tolerância ao Sal/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ácido Abscísico/metabolismo , Sódio/metabolismo , Plantas Geneticamente Modificadas , Estresse Salino/genéticaRESUMO
Vacuolar proteins play essential roles in plant physiology and development, but the factors and the machinery regulating their vesicle trafficking through the endomembrane compartments remain largely unknown. We and others have recently identified an evolutionarily conserved plant endosomal sorting complex required for transport (ESCRT)-associated protein apoptosis-linked gene-2 interacting protein X (ALIX), which plays canonical functions in the biogenesis of the multivesicular body/prevacuolar compartment (MVB/PVC) and in the sorting of ubiquitinated membrane proteins. In this study, we elucidate the roles and underlying mechanism of ALIX in regulating vacuolar transport of soluble proteins, beyond its conventional ESCRT function in eukaryotic cells. We show that ALIX colocalizes and physically interacts with the retromer core subunits Vps26 and Vps29 in planta. Moreover, double-mutant analysis reveals the genetic interaction of ALIX with Vps26 and Vps29 for regulating trafficking of soluble vacuolar proteins. Interestingly, depletion of ALIX perturbs membrane recruitment of Vps26 and Vps29 and alters the endosomal localization of vacuolar sorting receptors (VSRs). Taken together, ALIX functions as a unique retromer core subcomplex regulator by orchestrating receptor-mediated vacuolar sorting of soluble proteins.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Endossomos/metabolismo , Plantas/metabolismo , Transporte Proteico/fisiologia , Vacúolos/metabolismoRESUMO
BACKGROUND: To investigate the risk factors for cough after pulmonary resection. METHODS: The PubMed, Embase, Web of Science, ClinicalTrials.gov, and China National Knowledge Network databases were searched from inception to November 2022. The Q tests and I2 statistic were used to evaluate the heterogeneity. Odds ratios (OR) were combined using the inverse variance method. All statistical analyses were performed by RevMan 5.4.1. RESULTS: Nineteen studies with 4755 patients were included, the incidence of postoperative cough was 21.1%-55.8%. The results showed that young age [OR = 0.66, 95% CI (0.46, 0.96), p = 0.03], female sex [OR = 1.69, 95% CI (1.07, 2.66), p = 0.02], preoperative cough [OR = 5.96, 95% CI (2.58, 13.73), p < 0.01], right lobe operation [OR = 2.14, 95% CI (1.44, 3.19), p < 0.01], lobectomy [OR = 3.70, 95% CI (1.73, 7.90), p < 0.01], subcarinal lymph node dissection [OR = 3.45, 95% CI (1.86, 6.39), p < 0.01], mediastinal lymph node removal [OR = 3.49, 95% CI (2.07, 5.89), p < 0.01], closure of bronchial stump with stapler [OR = 5.19, 95% CI (1.79, 15.07), p < 0.01], peritracheal lymph node resection [OR = 3.05, 95%CI (1.40,6.64), p < 0.01], postoperative acid reflux [OR = 11.07, 95%CI (4.38,28.02), p < 0.01] were independent risk factors for cough after pulmonary resection. CONCLUSIONS: Young age, female sex, preoperative cough, right lobe operation, lobectomy, subcarinal lymph node dissection, mediastinal lymph node removal, closure of bronchial stump with stapler, peritracheal lymph node resection, and postoperative acid reflux are independent risk factors for cough after pulmonary resection.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Feminino , Humanos , Carcinoma Pulmonar de Células não Pequenas/patologia , Tosse/epidemiologia , Tosse/etiologia , Tosse/patologia , Neoplasias Pulmonares/patologia , Excisão de Linfonodo/métodos , Linfonodos/patologia , Estudos Retrospectivos , Fatores de Risco , MasculinoRESUMO
Skin appendages in vertebrates have individual morphological differences, but share the same evolutionary origin. In this study, we used Megalobrama amblycephala as a fish model to study the developmental regulation mechanism of a common skin appendage in fish: scales. By combining in-toto live imaging method and transcriptomic analysis during the scale development, we elucidated core features of scale patterning containing three distinct regions and experiencing four stages. Differentially expressed genes in skin tissues at the initial site before and after scale development were analyzed and some key regulatory genes (Wnt3, Wnt6, Fgf8, Fgf10, Fgf16, Fgfr1a, Ihhb and BMP6) which are crucial for scale morphogenesis were selected. This study provides a strong reference for further exploration of the function of genes related to the molecular regulation mechanism of scale development in M. amblycephala, as well as in other fishes.
Assuntos
Cyprinidae , Cipriniformes , Animais , Cyprinidae/genética , Cipriniformes/genética , Perfilação da Expressão Gênica , Morfogênese/genéticaRESUMO
PURPOSES: To develop a nomogram model for distinguishing benign from malignant ampullary lesions more intuitive and accurate. MATERIALS AND METHODS: A total of 124 patients with periampullary lesions from January 2016 to June 2020 were enrolled in this retrospective study. Their clinical information, ultrasound (US), dual contrast-enhanced ultrasound (DCEUS) and MRI image features were used for research. Twenty features were collected in our study. Random forest was used to select the first five most important indicators to construct the prediction model. RESULTS: Patients' age, common bile duct (CBD) diameter, the shape, vascularity, and boundary of lesion, lesion size with or without enlarged after CEUS, the enhancement patterns of arterial phase, the washout patterns of venous phase, CEUS diagnosis, and MRI diagnosis were statistically significant (p < 0.05). After screening for statistically significant indicators by random forest, the first five most important indicators were age, CBD diameter, the enhancement patterns of arterial phase, the washout patterns of venous phase, lesion size with or without enlarged after CEUS, which were used to construct nomogram. The area under curves (AUC) and 95% confidence intervals (CI) for nomogram, MRI + MRCP + DCEUS, DCEUS, MRI + MRCP were 0.98(0.94-1.00), 0.91(0.84-0.97), 0.89(0.80-0.98), 0.68(0.60-0.77), respectively. The sensitivity and specificity were 100.00% and 84.62% for nomogram, 88.29% and 92.31% for MRI + MRCP+DCEUS, 86.49% and 92.31% for DCEUS, 51.35%, and 100.00% for MRI + MRCP. CONCLUSIONS: We combined clinical indicators, gray-scale ultrasound characteristics, and CEUS characteristics to build the nomogram, which can be intuitively and accurately used for preoperative malignant prediction of ampullary lesion patients, worthy of clinical generalizability and application.
Assuntos
Meios de Contraste , Nomogramas , Humanos , Imageamento por Ressonância Magnética/métodos , Estudos Retrospectivos , Ultrassonografia/métodosRESUMO
Glycogen synthase kinase 3 (GSK3) proteins play key roles in brassinosteroid (BR) signaling during plant growth and development by phosphorylating various substrates. However, how GSK3 protein stability and activity are themselves modulated is not well understood. Here, we demonstrate in vitro and in vivo that C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 (OsCPL3), a member of the RNA Pol II CTD phosphatase-like family, physically interacts with OsGSK2 in rice (Oryza sativa). OsCPL3 expression was widely detected in various tissues and organs including roots, leaves and lamina joints, and was induced by exogenous BR treatment. OsCPL3 localized to the nucleus, where it dephosphorylated OsGSK2 at the Ser-222 and Thr-284 residues to modulate its protein turnover and kinase activity, in turn affecting the degradation of BRASSINAZOLE-RESISTANT 1 (BZR1) and BR signaling. Loss of OsCPL3 function resulted in higher OsGSK2 abundance and lower OsBZR1 levels, leading to decreased BR responsiveness and alterations in plant morphology including semi-dwarfism, leaf erectness and grain size, which are of fundamental importance to crop productivity. These results reveal a previously unrecognized role for OsCPL3 and add another layer of complexity to the tightly controlled BR signaling pathway in plants.
Assuntos
Brassinosteroides , Oryza , Brassinosteroides/metabolismo , Regulação da Expressão Gênica de Plantas , Quinase 3 da Glicogênio Sintase/genética , Quinase 3 da Glicogênio Sintase/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais/genéticaRESUMO
BACKGROUND: Neutrophil-lymphocyte count ratio (NLCR) has been reported as better indicator of bacteremia than procalcitonin (PCT), and more precise predictor of mortality than C-reactive protein (CRP) under various medical conditions. However, large controversy remains upon this topic. To address the discrepancy, our group has compared the efficiency of NLCR with conventional inflammatory markers in predicting the prognosis of critical illness. METHODS: We performed a multi-center retrospective cohort study involving 536 ICU patients with outcomes of survival, 28- and 7-day mortality. NLCR was compared with conventional inflammatory markers such as PCT, CRP, serum lactate (LAC), white blood cell, neutrophil and severity score APACHE II (Acute Physiology and Chronic Health Evaluation II) to evaluate the potential outcomes of critical illness. Then, receiver operating characteristics (ROC) curves were constructed to assess and compare each marker's sensitivity and specificity respectively. RESULTS: NLCR values were not different between survival and mortality groups. Meanwhile, remarkable differences were observed upon APACHE II score, CRP, PCT and LAC levels between survival and death groups. ROC analysis revealed that NLCR was not competent to predict prognosis of critical illness. The AUROCs of conventional markers such as CRP, PCT, LAC and APACHE II score were more effective in predicting 28- and 7-day mortality. CONCLUSIONS: NLCR is less reliable than conventional markers CRP, PCT, LAC and APACHE II score in assessing severity and in predicting outcomes of critical illness.
Assuntos
APACHE , Inflamação/sangue , Contagem de Linfócitos , Sepse , Adulto , Humanos , Unidades de Terapia Intensiva , Neutrófilos/citologia , Prognóstico , Estudos RetrospectivosRESUMO
BACKGROUND: The relationship between biomarkers and hospital-acquired pneumonia (HAP) is understudied, especially in severe cases admitted to the intensive care unit (ICU). Compared with community-acquired pneumonia (CAP), HAP might have different traits regarding biomarkers due to the previous history in hospitals. METHODS: A total of 593 adult patients were enrolled in this retrospective cohort study to determine the neutrophil/lymphocyte count ratio (NLCR), procalcitonin (PCT), C-reactive protein (CRP) and serum lactate level upon admission to the ICU. According to diagnosis, patients were divided into two groups: non-infection and HAP. Discriminant analysis was performed based on better outcomes of diagnostic performance and severity evaluation. The diagnostic performance of each individual biomarker was assessed by constructing receiver operating characteristic (ROC) curves and calculating the area under each ROC curve (AUROC). Multivariable analysis was also applied to determine the most appropriate prognostic factors. RESULTS: NLCR, PCT and CRP were markedly different between the non-infection and HAP groups. NLCR had a worse ability to discriminate severe infection (AUROC 0.626; 95% CI 0.581-0.671) than conventional markers such as CRP (0.685, 95% CI 0.641-0.730) and PCT (0.661, 95% CI 0.615-0.707). In addition, the AUROC of composite biomarkers, especially the combination of NLCR, CRP and WBC, was significantly greater than that of any single biomarker. CONCLUSIONS: NLCR was not comparable to conventional single biomarkers, such as CRP and PCT, for diagnosing or evaluating the severity of HAP. Composite biomarkers that have good accessibility, especially the combination of NLCR, CRP and WBC, could help with early diagnosis and severity evaluation.
Assuntos
Proteína C-Reativa/análise , Pneumonia Associada a Assistência à Saúde/sangue , Pró-Calcitonina/sangue , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Diagnóstico Precoce , Feminino , Pneumonia Associada a Assistência à Saúde/diagnóstico , Humanos , Unidades de Terapia Intensiva , Contagem de Leucócitos , Modelos Logísticos , Contagem de Linfócitos , Linfócitos , Masculino , Pessoa de Meia-Idade , Neutrófilos/citologia , Curva ROC , Estudos RetrospectivosRESUMO
Since December 2019, the medical staff fighting against COVID-19 frequently reported the device-related pressure injury (DRPI) caused by personal protective equipment (PPE). We conducted a cross-sectional survey online to investigate the prevalence and characteristics of DRPI among medical staff. Univariate and multivariate logistic regression analyses were employed to explore the risk factors associated with DRPI. A total of 4308 participants were collected and 4306 participants were valid from 161 hospitals in China. The overall prevalence of DRPI caused by PPE among medical staff was 30.03% (95% CI 28.69%-31.41%). The prevalence of male was more than that of female (42.25%, 95% CI 37.99-46.51% vs 26.36%, 95% CI 26.93-29.80%, P < .001).The categories were mainly stages 1 and 2, and the common anatomical locations were nose bridge, cheeks, ears, and forehead. Logistic regression analysis revealed that the risk factors were sweating (OR = 43.99, 95% CI 34.46-56.17), male (OR = 1.50, 95% CI 1.12-1.99), level 3 PPE (OR = 1.44, 95% CI 1.14-1.83), and longer wearing time (OR = 1.28, 95% CI 0.97-1.68). The prevalence of DRPI was high among medical staff wearing PPE against COVID-19, and the risk factors were sweating, male, wearing level 3 PPE, and longer wearing time. Comprehensive preventive interventions should be taken.
Assuntos
COVID-19/prevenção & controle , Corpo Clínico Hospitalar , Recursos Humanos de Enfermagem Hospitalar , Traumatismos Ocupacionais/etiologia , Equipamento de Proteção Individual/efeitos adversos , Úlcera por Pressão/etiologia , Adulto , COVID-19/transmissão , China/epidemiologia , Estudos Transversais , Transmissão de Doença Infecciosa/prevenção & controle , Traumatismos Faciais/etiologia , Feminino , Humanos , Masculino , Prevalência , Fatores de Risco , Fatores Sexuais , Inquéritos e Questionários , Sudorese , Fatores de TempoRESUMO
INTRODUCTION AND AIM: Acute-on-chronic liver failure (ACLF) is a syndrome with high short-term mortality, and predicting the prognosis is challenging. This study aimed to compare the performance of neutrophil gelatinase-associated lipocalin (NGAL) and cystatin C (CysC) in predicting the 90-day mortality in patients with hepatitis B virus (HBV)-associated ACLF (HBV-ACLF). MATERIALS AND METHODS: This prospective, observational study enrolled 54 patients with HBV-ACLF. The serum NGAL and CysC levels were determined. A multivariate logistic regression analysis was used to analyze the independent risk factors of mortality. RESULTS: Serum NGAL, but not CysC, was found to significantly correlate with the total bilirubin, international normalized ratio, and model for end-stage liver disease (MELD). Serum NGAL [odds ratio (OR), 1.008; 95% confidence interval (CI), 1.004-1.012; P < 0.01], but not CysC, was an independent risk factor for developing hepatorenal syndrome. Moreover, NGAL (OR, 1.005; 95% CI, 1.001-1.010; P < 0.01) along with the MELD score was independently associated with the overall survival in patients with HBV-ACLF. Patients with HBV-ACLF were stratified into two groups according to the serum NGAL level at baseline (low risk: <217.11 ng/mL and high risk: ≥ 217.11 ng/mL). The 90-day mortality rate was 22.73% (5/22) in the low-risk group and 71.88% (23/32) in the high-risk group. Moreover, NGAL, but not CysC, significantly improved the MELD score in predicting the prognosis of HBV-ACLF. CONCLUSION: The serum NGAL might be superior to CysC in predicting the prognosis of HBV-ACLF with the normal creatinine level.
Assuntos
Insuficiência Hepática Crônica Agudizada/sangue , Cistatina C/sangue , Lipocalina-2/sangue , Insuficiência Hepática Crônica Agudizada/diagnóstico , Insuficiência Hepática Crônica Agudizada/mortalidade , Adulto , Biomarcadores/sangue , China/epidemiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Estudos Prospectivos , Fatores de Risco , Taxa de Sobrevida/tendênciasRESUMO
Interleukin (IL)-22, as a member of the interleukin (IL)-10 family, is an important mediator between the immune cells and epithelial tissues during infection and inflammation. This study reported the characterization and mRNA expression patterns of Pf_IL-22 gene and its cell surface-associated receptors Pf_IL-22RA1 and soluble Pf_IL-22RA2 genes in yellow catfish (Pelteobagrus filvidraco). The open reading frames (ORFs) of the Pf_IL-22, Pf_IL-22RA1 and Pf_IL-22RA2 genes were 546 bp, 1740 bp and 690 bp in length, encoding 181, 579 and 229 amino acids, respectively. Alignments of the deduced amino acid sequences present that the Pf_IL-22 has a conserved IL-10 family signature motif, and the Pf_IL-22RA1 and Pf_IL-22RA2 have two conserved fibronectin type-III domains. Quantitative real-time PCR (qPCR) analyses showed that the Pf_IL-22 and Pf_IL-22RA1 mRNAs were highly expressed in mucosal tissues such as the fin, gill, intestine, skin mucus and stomach, and were weakly expressed in the kidney, liver and head kidney of adult yellow catfish, indicating that the Pf_IL-22 transcripts may be mainly produced by mucosal immune cells/tissues in healthy yellow catfish. The mRNA expression levels of the Pf_IL-22RA2 gene were high in the muscle and liver, and were relatively low in the spleen and kidney. The mRNA expression levels of the Pf_IL-22 and its two receptor genes were significantly up-regulated in both mucosal tissues (gill, hindgut, and skin mucus) and systemic immune tissues (spleen, head kidney and blood) after Edwardsiella ictaluri challenge. These results indicated that the Pf_IL-22 and its two receptors genes might play an important role in the innate immune defense against bacterial invasion.
Assuntos
Peixes-Gato , Edwardsiella ictaluri , Infecções por Enterobacteriaceae , Doenças dos Peixes , Proteínas de Peixes , Interleucinas , Receptores de Interleucina , Nadadeiras de Animais/metabolismo , Animais , Peixes-Gato/genética , Peixes-Gato/imunologia , Edwardsiella ictaluri/imunologia , Infecções por Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Proteínas de Peixes/metabolismo , Mucosa Gástrica/metabolismo , Brânquias/metabolismo , Rim Cefálico/metabolismo , Interleucinas/genética , Interleucinas/imunologia , Interleucinas/metabolismo , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Músculos/metabolismo , RNA Mensageiro/metabolismo , Receptores de Interleucina/genética , Receptores de Interleucina/imunologia , Receptores de Interleucina/metabolismo , Pele/metabolismo , Baço/metabolismo , Interleucina 22RESUMO
BACKGROUND: Glabridin (GLA), a major component extracted from licorice root, has anti-inflammatory and antioxidant activities, but few studies report its mechanism of inhibition of angiogenesis. This study was an extension of our previous work, which demonstrated that GLA suppressed angiogenesis in human breast cancer (MDA-MB-231 and Hs-578T) cells. Breast cancer is one of the most common malignant diseases in females worldwide, and the major cause of mortality is metastasis that is primarily attributed to angiogenesis. Thus, anti-angiogenesis has become a strategy for the treatment of breast cancer. METHODS: Cell viability of different concentration treatment groups were detected by Cell Counting Kit-8 assay. The expression of several related genes in the Wnt1 signaling pathway in MDA-MB-231 and Hs-578T cells treated with GLA were measured at both the transcription and translation levels using quantitative real-time PCR analyses and western blotting. Immunofluorescence assay analyzed the nuclear translocation of ß-catenin. The microRNA-inhibitor was used to knockdown microRNA-148a (miR-148a) expression. Angiogenic potentials of breast cancer cells were analyzed by enzyme-linked immunosorbent assay (ELISA) and tube formation in vitro. RESULTS: GLA attenuated angiogenesis by the suppression of miR-148a-mediated Wnt/ß-catenin signaling pathway in two human breast cancer cell lines (MDA-MB-231 and Hs-578T). GLA also upregulated the expression of miR-148a in a dose-dependent manner, miR-148a, which could directly target Wnt-3'-untranslated regions (UTRs), and decreased the expression of Wnt1, leading to ß-catenin accumulation in the membranes from the cytoplasm and nucleus. Downregulation of miR-148a contributed to the reduction of GLA-induced suppression of the Wnt/ß-catenin signaling pathway, the angiogenesis and vascular endothelial grow factor (VEGF) secretion. CONCLUSIONS: Our study identified a molecular mechanism of the GLA inhibition of angiogenesis through the Wnt/ß-catenin signaling pathway via miR-148a, suggesting that GLA could serve as an adjuvant chemotherapeutic agent for breast cancer.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/metabolismo , Isoflavonas/farmacologia , MicroRNAs/genética , Neovascularização Patológica/metabolismo , Fenóis/farmacologia , Via de Sinalização Wnt/genética , Linhagem Celular Tumoral , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , MicroRNAs/metabolismo , Transdução de Sinais/genéticaRESUMO
Hirschsprung disease (HSCR) is a congenital disorder caused by the defective function of the embryonic enteric neural crest. The impaired migration of embryonic enteric neural crest plays an important role in the pathogenesis of this disease. Recent studies showed that the ARP2/3 complex and RAC isoforms had effects on actin cytoskeleton remodelling, which contributes to migration. Moreover, some regulatory relationships were identified between ARP2/3 complex and RAC isoforms. Although microRNAs (miRNAs) have been known to modulate target gene expression on the post-transcriptional level, little is known about the regulation among miRNAs, ARP2/3 complex and RAC isoforms. Here, we report that down-regulation of ARP2 and ARP3, two main subunits of ARP2/3 complex, suppressed migration and proliferation in 293T and SH-SY5Y cell lines via the inhibition of RAC1 and RAC2. Meanwhile, as the target genes, ARP2 and ARP3 are reduced by increased miR-24-1* and let-7a*, respectively, in 70 HSCR samples as compared with 74 normal controls. Co-immunoprecipitation showed that aberrant reduction in ARP2 and ARP3 could weaken the function of ARP2/3 complex. Our study demonstrates that the miR-24-1*/let-7a*-ARP2/3 complex-RAC isoforms pathway may represent a novel pathogenic mechanism for HSCR.
Assuntos
Complexo 2-3 de Proteínas Relacionadas à Actina/metabolismo , Movimento Celular , Doença de Hirschsprung/genética , Doença de Hirschsprung/patologia , MicroRNAs/metabolismo , Proteínas rac de Ligação ao GTP/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo , Complexo 2-3 de Proteínas Relacionadas à Actina/genética , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células , Demografia , Regulação para Baixo/genética , Feminino , Humanos , Imunoprecipitação , Lactente , Masculino , MicroRNAs/genética , Modelos Biológicos , Isoformas de Proteínas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regulação para Cima/genética , Proteína RAC2 de Ligação ao GTPRESUMO
OBJECTIVE: To investigate the effects of initiative and passive perioperative function exercises on hidden blood loss (HBL). METHODS: Two hundreds and thirty elderly patients with hip fractures aging from 67 to 87 years (average age of 73.6 years) who underwent total hip replacement were included. By the intensity and the manner of perioperative function exercises, patients were divided into four groups: little initiative function exercises group (group A, n=51), little initiative and passive function exercises group (group B, n=54), normal initiative function exercises group (group C, n=65), normal initiative and passive function exercises group (group D, n=60). The true total blood loss, HBL and their proportion on the original blood volume and total blood loss was calculated depending on height, weight, intra-operative blood loss, post-operative blood loss, pre- and post-operative hematocrit, and blood transfused. According to the proportion of mean HBL on total blood loss, patients were divided into low HBL group and high HBL group. The data were analyzed by t test. RESULTS: The mean HBL was 517 ml, 41.9% of the total blood loss. Thereinto, the mean HBL was 695 ml in group A, 49.3% of the total blood loss, the prevalence of high HBL was 66.7% (34/51); the mean HBL was 625 ml in group B, 46.9% of the total blood loss, the prevalence of high HBL was 59.3% (32/54); the mean HBL was 446 ml in group C, 38.4% of the total blood loss, the prevalence of high HBL was 30.8% (20/65); the mean HBL was 346 ml in group D, 32.3% of the total blood loss, the prevalence of high HBL was 20.0% (12/60). Mean HBL, mean HBL/total blood loss, prevalence of high HBL were lower in group C than that in group A and group B (all P<0.05); and were lower in group D than that in group C (all P<0.05). The prevalence was 57.4% (132 cases) in low HBL group, and 42.6% (98 cases) in high HBL. The proportion of little initiative function exercises patients in high HBL group was obviously higher than that in low HBL group (P<0.05). CONCLUSIONS: The intensity and the manner of perioperative function exercises are strongly associated with the HBL in elderly patients with total hip replacement. The initiative combined with the passive function exercises could be effectively prevent and reduce the incidence of high HBL.
Assuntos
Artroplastia de Quadril , Perda Sanguínea Cirúrgica , Terapia por Exercício/efeitos adversos , Hemorragia Pós-Operatória , Idoso , Idoso de 80 Anos ou mais , Fraturas do Quadril/cirurgia , HumanosRESUMO
Recent studies have emphasized the important role of microRNA (miRNA) clusters and common target genes in disease progression. Despite the known involvement of the miR-192/215 family in many human diseases, its biological role in Hirschsprung disease (HSCR) remains undefined. In this study, we explored the role of the miR-192/215 family in the pathogenesis of HSCR. Quantitative real-time PCR and western blotting measured relative expression levels of miRNAs, mRNAs, and proteins in 80 HSCR patients and 77 normal colon tissues. Targets were evaluated by dual-luciferase reporter assays, and the functional effects of miR-192/215 on human 293T and SH-SY5Y cells were detected by the Transwell assay, CCK8 assay and flow cytometry. MiR-192/215 was significantly down-regulated in HSCR tissue samples, and their knockdown inhibited cell migration and proliferation in the human 293T and SH-SY5Y cell lines. Nidogen 1 (NID1) was confirmed as a common target gene of miR-192/215 by dual-luciferase reporter gene assay and its expression was inversely correlated with that of miR-192/215 in tissue samples and cell lines. Silencing of NID1 could rescue the extent of the suppressing effects by miR-192/215 inhibitor. The down-regulation of miR-192/215 may contribute to HSCR development by targeting NID1. We proposed the following cascade for the proposed mechanism of miR-192/215 in the pathogenesis of Hirschsprung disease (HSCR) by targeting Nidogen 1 (NID1). Aberrant expression of miR-192/215 inhibits cell migration and cell proliferation via NID1. We think the miR-192/miR-215/NID1 signaling pathway may play an important role in the pathogenesis of HSCR.
Assuntos
Doença de Hirschsprung/metabolismo , Doença de Hirschsprung/patologia , Glicoproteínas de Membrana/biossíntese , MicroRNAs/biossíntese , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Feminino , Células HEK293 , Doença de Hirschsprung/genética , Humanos , Lactente , Masculino , Glicoproteínas de Membrana/genética , MicroRNAs/genética , Transdução de Sinais/fisiologiaRESUMO
Long noncoding RNAs (lncRNAs) have been confirmed to be associated with various human diseases. However, whether they are associated with Hirschsprung disease (HSCR) progression remains unclear. In this study, we designed the experiment to explore the relationship between lncRNA HOTTIP and HOXA13, and their pathogenicity to HSCR. Quantitative real-time PCR and Western blot were performed to detect the levels of lncRNA, mRNAs, and proteins in colon tissues from 79 patients with HSCR and 79 controls. Small RNA interference transfection was used to study the function experiments in human 293T and SK-N-BE cell lines. The cell viability and activities were detected by the transwell assays, CCK8 assay, and flow cytometry, respectively. LncRNA HOTTIP and HOXA13 were significantly down-regulated in HSCR compared to the controls. Meanwhile, the declined extent of their expression levels makes sense between two main phenotype of HSCR. SiRNA-mediated knock-down of HOTTIP or HOXA13 correlated with decreased levels of each other and both reduced the cell migration and proliferation without affecting cell apoptosis or cell cycle. Our study demonstrates that aberrant reduction of HOTTIP and HOXA13, which have a bidirectional regulatory loop, may play an important role in the pathogenesis of HSCR.
Assuntos
Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Doença de Hirschsprung/patologia , Proteínas de Homeodomínio/fisiologia , RNA Longo não Codificante/fisiologia , Estudos de Casos e Controles , Linhagem Celular , Regulação para Baixo , Feminino , Proteínas de Homeodomínio/genética , Humanos , Lactente , Masculino , RNA Longo não Codificante/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: Previous studies suggested that cytochrome P450 participated in the tumor metastasis and migration. CYP2B6 also acts as an important enzyme which metabolize partially or primarily metabolism of drugs, environmental contaminants, and mutagens. The objective of this study was to investigate the influence of CYP2B6 polymorphism on susceptibility of Hirschsprung disease. METHODS: TaqMan assay was performed to determine the genotypes of CYP2B6 rs707265, rs1042389, rs2054675 in 262 cases and 290 control subjects. Logistic regression was used to assess the associations between these polymorphisms and HSCR. RESULTS: We observed a significant association of CYP2B6 rs707265 (G>A) polymorphism and HSCR susceptibility (p < 0.001). Besides, rs707265 A presented a significant risk of HSCR (p < 0.001). CONCLUSIONS: Our result suggested that CYP2B6 rs707265 modified the risk of HSCR.
Assuntos
Povo Asiático/genética , Citocromo P-450 CYP2B6/genética , Doença de Hirschsprung/genética , Polimorfismo Genético , Estudos de Casos e Controles , China/epidemiologia , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Doença de Hirschsprung/enzimologia , Doença de Hirschsprung/etnologia , Humanos , Modelos Logísticos , Razão de Chances , Fenótipo , Fatores de RiscoRESUMO
BACKGROUND: Hirschsprung disease (HSCR) is a rare multigenic congenital disorder characterized by the absence of the enteric ganglia. To date, single nucleotide polymorphisms (SNPs) in pre-miRNAs have been confirmed related with some diseases. Thus, we hypothesized that pre-miRNA polymorphisms might contribute to HSCR susceptibility. We investigated whether rs2910164 and rs11614913 of pre-miR-146a and pre-miR-196a2, are associated with HSCR. METHODS: Polymorphisms were genotyped using the Taqman method. Real-time PCR was used for detecting the expression level of miR-146a and its target gene ROBO1 in CC and GG genotypes. RESULTS: Significant differences were found in the genotype distribution of rs2910164 and rs11614913 polymorphism between HSCR cases and controls (p = 0.023 and 0.041, respectively). Furthermore, G allele of rs2910164 might increase the risk of HSCR (OR, 1.54; 95% CI, 1.06-2.23). Moreover, the expression level of miR-146a for homozygote GG was also higher than homozygote CC (p = 0.0193). In contrast, the expression level of its target gene ROBO1 predicted in bioinformatics for homozygote GG was much lower than homozygote CC (p = 0.0096). CONCLUSIONS: Our results showed that the polymorphism rs2910164 in pre-miR-146a might alter the production of mature miR-146a and then down-regulate the target gene ROBO1, which plays an important role in pathogenesis of HSCR.
Assuntos
Povo Asiático/genética , Predisposição Genética para Doença/genética , Doença de Hirschsprung/genética , MicroRNAs/genética , Proteínas do Tecido Nervoso/genética , Receptores Imunológicos/genética , Feminino , Genótipo , Humanos , Lactente , Masculino , Proteínas do Tecido Nervoso/biossíntese , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Receptores Imunológicos/biossíntese , Fatores de Risco , Proteínas RoundaboutRESUMO
Agrobacterium-mediated transient expression is a flexible and efficient technique for introducing genes into plants, allowing for rapid and temporary gene expression. Agroinfiltration of Arabidopsis seedlings is a newly developed Agrobacterium-based transient expression system. The expression of target genes and the localization of relevant proteins can be observed within 3 days using this method. In this chapter, we present the detailed protocol for transient transformation in Arabidopsis thaliana seedlings utilizing vacuum infiltration of Agrobacterium. This procedure enables rapid and temporary gene expression by introducing exogenous DNA into Arabidopsis seedlings, particularly in easily accessible tissues such as cotyledons. This protocol provides a detailed description of experimental procedures, including Arabidopsis seedlings cultivation, the preparation of Agrobacterium suspensions, and subsequent steps leading to confocal microscope observation. Through this protocol, researchers can efficiently investigate gene function and subcellular localization in Arabidopsis cotyledons within 8 days in total.
Assuntos
Arabidopsis , Plântula , Arabidopsis/genética , Arabidopsis/metabolismo , Plântula/genética , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Vácuo , Cotilédone/genética , Cotilédone/metabolismo , Transformação Genética , Expressão Gênica , Plantas Geneticamente Modificadas/genética , Agrobacterium/genética , Regulação da Expressão Gênica de Plantas , Microscopia ConfocalRESUMO
Background: Numerous studies have consistently demonstrated a decline in sleep quality during the COVID-19 pandemic. The primary objective of this study is to explore the impact of engaging with pertinent epidemic information through the media amid the COVID-19 crisis on individuals' sleep quality and the underlying mechanisms through which this influence operates. Methods: An online cross-sectional study design was employed. A total of 1,063 British adults (36.2% males; M age = 38.85, SD age = 13.36, ranging from 18 to 77 years old) participated in the study and completed our questionnaires, which included media usage frequency during the pandemic, the 10-item Kessler Psychological Distress Scale (K10), the Insomnia Severity Index (ISI), and the Ten-item Personality Inventory (TIPI). Results: Pearson's correlation analyses indicated that there was no significant correlation between COVID-19-related traditional media use (television, radio, newspaper) and psychological distress or sleep quality. However, exposure to information related to COVID-19 through new media use (Facebook, Tik Tok, Twitter) was correlated with greater psychological distress and poorer sleep quality. A moderated mediation analysis showed that psychological distress fully mediated the relationship between new media use and poor sleep, which was moderated by age, with the association between psychological distress and poor sleep quality being stronger among older adults. Conclusion: Exposure to information of COVID-19 via new (but not traditional) media use deteriorated sleep quality through greater psychological distress, and this relationship was stronger among older adults.