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INTRODUCTION: The organisational silence of nursing teams has received increasing attention from managers. Chinese nurses have a relatively high score for organisational silence, and male nurses score higher than female nurses. Lack of professional empathy, high pressure in the work environment, and traditional Chinese cultural factors suggest that Chinese male nurses' experiences of and reasons for organisational silence are complex and unique. Taking male nurses in the emergency department as an example, this study explores the experience and meaning of male nurses' organisational silence and provides ideas for nursing managers to understand the silence of male nurses. METHODS AND ANALYSIS: An interpretative phenomenological approach underpins the study design. In this study, the purposive sampling method will be used to select male nurses who meet the inclusion criteria with maximum differentiation as a strategy. Face-to-face semistructured interviews and Van Manen analysis methods will be used for data collection and analysis. ETHICS AND DISSEMINATION: The study was approved by the Ethics Committee of the First Affiliated Hospital of Zhejiang Chinese Medical University (ethical approval ID: 2019-KL-036-01). Participants will provide informed consent, will be able to withdraw at any time and will have their contributions kept confidential. The findings of the study will be shared with relevant stakeholders and disseminated in conference presentations and journal publications. TRIAL REGISTRATION NUMBER: Chinese Clinical Trial Registry (ChiCTR2100047057).
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Serviço Hospitalar de Emergência , Enfermeiros , Equipe de Enfermagem , China , Humanos , Masculino , Enfermeiros/psicologia , Equipe de Enfermagem/organização & administraçãoRESUMO
Background: In the perioperative management of Total Knee Arthroplasty (TKA), postoperative fever has always been a concern. Current research focuses on infectious fever, and there is no relevant research on the occurrence of non-infectious fever (NIF) and its risk factors. Hence, the aim of this study was to clarify the risk factors for NIF after TKA, and construct an easy-to-use nomogram. Methods: A retrospective cohort study was conducted. Consecutive patients undergoing primary unilateral TKA were divided into the non-infectious fever group and the control group. Clinicopathological characters were collected from electronic medical records. Univariate Logistic regression was used to analyze the related independent risk factors. The optimal threshold for each selected factor and combined index was determined when the Youden index achieved the highest value. And the predictive nomogram was developed by these independent factors. Results: Ultimately, 146 patients were included in this study. Of them, 57 (39.04%) patients experienced NIF. Results of the univariable logistic regression analysis indicated that intraoperative blood loss (OR, 1.002; 95% CI, 1.000-1.0004), postoperative drainage fluid volume (OR, 1.003; 95% CI, 1.001-1.006) and frequency of blood transfusion (n = 1; OR, 0.227; 95% CI, 0.068-0.757) were independent risk factors of NIF occurrence. The predictive nomogram that incorporated the above independent risk factors was developed, and it yielded an areas under the curves (AUC) of 0.731 (95% CI: 0.651-0.801; P < 0.0001) with 54.39% sensitivity and 82.02% specificity. Conclusions: Non-infectious fever after TKA prolongs the time of antibiotic use and hospital stay. Our results demonstrated that the nomogram may facilitate to predict the individualized risk of NIF occurrence within 7-day by intraoperative blood loss, postoperative drainage fluid volume and frequency of blood transfusion.
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BACKGROUND: The outcome of surgically resected, apparently localized, clear cell renal carcinoma (ccRCC) is uncertain. OBJECTIVE: To evaluate if cell cycle progression (CCP) gene expression can predict future metastasis. METHODS: Pathologic T2a-T3b tumors at University of Iowa were reviewed. Patients with known or suspected metastasis, lymph node involvement or who received neoadjuvant or adjuvant radiation, chemotherapy or immunotherapy were excluded. Case and control cohorts were defined as those who did or did not develop metastatic disease within 5 years. Measured levels of 31 cell cycle genes and 15 control genes from the tumor were calculated as a CCP score. Additionally, gene expression data for a separate ccRCC cohort was downloaded from The Cancer Genome Atlas (TCGA). RESULTS: Univariate analysis of 26 cases and 38 controls revealed that the CCP score predicted progression to metastasis (OR 2.65, p = 0.0091). In multivariate logistic regression modeling, CCP expression remained a significant independent predictor for progression (p = 0.026). The CCP score was also significantly associated with distant metastasis in the TCGA renal cancer cohort in both univariate (p = 1.0 × 10-9) and multivariate (p = 5.6 × 10-3) analysis. CONCLUSION: The CCP score has prognostic value in predicting metastatic progression after resection of organ-confined ccRCC.
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Biomarcadores Tumorais/genética , Carcinoma de Células Renais/secundário , Proteínas de Ciclo Celular/genética , Ciclo Celular/genética , Neoplasias Renais/patologia , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/cirurgia , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Neoplasias Renais/genética , Neoplasias Renais/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , Reação em Cadeia da Polimerase em Tempo Real , Estudos RetrospectivosRESUMO
Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) negatively regulates the phosphoinositide-3-kinase (PI3K) signaling pathway. In colorectal cancer (CRC), observed frequencies of loss of PTEN expression, concordant expression in primary tumors and metastases, and the association of PTEN status with outcome vary markedly by detection method. We determined the degree to which PTEN expression is consistent in 70 matched human CRC primaries and liver metastases using a validated immunohistochemistry assay. We found loss of PTEN expression in 12.3% of assessable CRC primaries and 10.3% of assessable liver metastases. PTEN expression (positive or negative) was concordant in 98% of matched colorectal primaries and liver metastases. Next we related PTEN status to mutations in RAS and PI3K pathway genes (KRAS, NRAS, BRAF , and PIK3CA) and to overall survival (OS). PTEN expression was not significantly associated with the presence or absence of mutations in RAS or PI3K pathway genes. The median OS of patients whose tumors did not express PTEN was 9 months, compared to 49 months for patients whose tumors did express PTEN (HR = 6.25, 95% confidence intervals (CI) (1.98, 15.42), P = 0.0017). The association of absent PTEN expression with increased risk of death remained significant in multivariate analysis (HR = 6.31, 95% CI (2.03, 17.93), P = 0.0023). In summary, PTEN expression was consistent in matched CRC primaries and in liver metastases. Therefore, future investigations of PTEN in metastatic CRC can use primary tumor tissue. In patients with liver-only metastases, loss of PTEN expression predicted poor OS.
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Neoplasias Colorretais/genética , Neoplasias Colorretais/mortalidade , Expressão Gênica , PTEN Fosfo-Hidrolase/genética , Idoso , Classe I de Fosfatidilinositol 3-Quinases , Neoplasias Colorretais/patologia , Neoplasias Colorretais/terapia , Feminino , Genes ras , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Mutação , Gradação de Tumores , Metástase Neoplásica , Estadiamento de Neoplasias , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/genética , Prognóstico , Proteínas Proto-Oncogênicas B-raf/genéticaRESUMO
The identification and validation of the targets of active compounds identified in cell-based assays is an important step in preclinical drug development. New analytical approaches that combine drug affinity pull-down assays with mass spectrometry (MS) could lead to the identification of new targets and druggable pathways. In this work, we investigate a drug-target system consisting of ampicillin- and penicillin-binding proteins (PBPs) to evaluate and compare different amino-reactive resins for the immobilization of the affinity compound and mass spectrometric methods to identify proteins from drug affinity pull-down assays. First, ampicillin was immobilized onto various amino-reactive resins, which were compared in the ampicillin-PBP model with respect to their nonspecific binding of proteins from an Escherichia coli membrane extract. Dynal M-270 magnetic beads were chosen to further study the system as a model for capturing and identifying the targets of ampicillin, PBPs that were specifically and covalently bound to the immobilized ampicillin. The PBPs were identified, after in situ digestion of proteins bound to ampicillin directly on the beads, by using either one-dimensional (1-D) or two-dimensional (2-D) liquid chromatography (LC) separation techniques followed by tandem mass spectrometry (MS/MS) analysis. Alternatively, an elution with N-lauroylsarcosine (sarcosyl) from the ampicillin beads followed by in situ digestion and 2-D LC-MS/MS analysis identified proteins potentially interacting noncovalently with the PBPs or the ampicillin. The in situ approach required only little time, resources, and sample for the analysis. The combination of drug affinity pull-down assays with in situ digestion and 2-D LC-MS/MS analysis is a useful tool in obtaining complex information about a primary drug target as well as its protein interactors.
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Ampicilina/metabolismo , Sistemas de Liberação de Medicamentos , Proteínas de Ligação às Penicilinas/metabolismo , Espectrometria de Massas/métodos , Microesferas , Sarcosina/análogos & derivados , Sarcosina/química , Sefarose/análogos & derivadosRESUMO
Activation of cyclic nucleotide dependent signaling pathways leads to relaxation of smooth muscle, alterations in the cytoskeleton of cultured cells, and increases in the phosphorylation of HSP20. To determine the effects of phosphorylated HSP20 on the actin cytoskeleton, phosphopeptide analogs of HSP20 were synthesized. These peptides contained 1) the amino acid sequence surrounding the phosphorylation site of HSP20, 2) a phosphoserine, and 3) a protein transduction domain. Treatment of Swiss 3T3 cells with phosphopeptide analogs of HSP20 led to loss of actin stress fibers and focal adhesion complexes as demonstrated by immunocytochemistry, interference reflection microscopy, and biochemical quantitation of globular-actin. Treatment with phosphopeptide analogs of HSP20 also led to dephosphorylation of the actin depolymerizing protein cofilin. Pull-down assays demonstrated that 14-3-3 proteins associated with phosphopeptide analogs of HSP20 (but not peptide analogs in which the serine was not phosphorylated). The binding of 14-3-3 protein to phosphopeptide analogs of HSP20 prevented the association of cofilin with 14-3-3. These data suggest that HSP20 may modulate actin cytoskeletal dynamics by competing with the actin depolymerizing protein cofilin for binding to the scaffolding protein 14-3-3. Interestingly, the entire protein was not needed for this effect, suggesting that the association is modulated by phosphopeptide motifs of HSP20. These data also suggest the possibility that cyclic nucleotide dependent relaxation of smooth muscle may be mediated by a thin filament (actin) regulatory process. Finally, these data suggest that protein transduction can be used as a tool to elucidate the specific function of peptide motifs of proteins.
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Citoesqueleto/metabolismo , Proteínas de Choque Térmico/metabolismo , Fosfopeptídeos/metabolismo , Fosfoproteínas/metabolismo , Proteínas 14-3-3/metabolismo , Células 3T3/química , Células 3T3/metabolismo , Fatores de Despolimerização de Actina , Actinina/metabolismo , Actinas/metabolismo , Animais , Sítios de Ligação , Linhagem Celular , Proteínas do Citoesqueleto/metabolismo , Adesões Focais/metabolismo , Proteínas de Choque Térmico HSP20 , Proteínas de Choque Térmico/química , Camundongos , Proteínas dos Microfilamentos/metabolismo , Paxilina , Peptídeos/química , Peptídeos/metabolismo , Fosfopeptídeos/síntese química , Fosfoproteínas/química , Fosforilação , Fosfosserina/metabolismo , Estrutura Terciária de Proteína , Vinculina/metabolismoRESUMO
In this study, a two-dimensional LC-MALDI-TOF/TOF method has been developed for analyzing protein complexes. In our hands, the method has proven to be an excellent strategy for the analysis of protein complexes isolated in pull-down experiments. This is in part because the preservation of the chromatographic separation on a MALDI target yields an "unlimited" amount of time to obtain MS/MS spectra, making it possible to probe more deeply into complex samples. A brief statistical analysis was performed on the data obtained from the LC-MALDI-TOF/TOF system in order to better understand peptide fragmentation patterns under high-energy collision conditions. These statistical analyses provided some insight into how to evaluate the quality and accuracy of the database search results derived from the TOF/TOF-based analysis. The potential of the method was demonstrated by the successful identification of all the known penicillin-binding proteins in E. coli isolated using a drug-based pull-down with ampicillin as the bait. The performance of the LC-MALDI-TOF/TOF system was compared with that of an equivalent 2D LC-ESI-MS/MS approach, in the analysis of a protein bait-based pull-down. Regardless of the number of peptides identified in the ESI versus MALDI approach, the two approaches were found to be complementary. When the data is merged at the peptide level, the combined result gives higher Mascot scores and an overall higher confidence in protein identification than with either approach alone.