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OBJECTIVE: To develop a comprehensive compliance assessment scale for postoperative visual function rehabilitation in children with congenital cataracts and to assess its reliability and validity. METHOD: Drawing on the Interactive Model of Health Behavior, we conducted a literature review and semi-structured interviews to create a pool of 36 items. The items underwent rigorous evaluation through the Delphi method, face validity checks, and item analysis, leading to a reduction to 18 items. To assess the scale's reliability and validity, we collected data from 225 parents of children with congenital cataracts. We employed SPSS version 25.0 for data analysis and evaluated construct validity using exploratory factor analysis, content validity, internal consistency reliability, and test-retest reliability. RESULTS: The compliance scale for postoperative visual function rehabilitation in children with congenital cataracts comprises 5 dimensions and 18 items. Exploratory factor analysis extracted 5 common factors, with a cumulative variance contribution rate of 68.178%. Item-level content validity index ranged from 0.730 to 1.000, and the content validity index of the scale was 0.963. The total Cronbach's alpha coefficient, split-half reliability, and test-retest reliability of the scale were 0.855, 0.778, and 0.859, respectively. CONCLUSIONS: The compliance assessment scale for postoperative visual function rehabilitation in children with congenital cataracts demonstrates acceptable reliability and validity. It serves as a valuable reference for developing standardized nursing programs for these children in clinical practice.
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Catarata , Humanos , Catarata/congênito , Catarata/fisiopatologia , Catarata/reabilitação , Feminino , Masculino , Pré-Escolar , Reprodutibilidade dos Testes , Criança , Extração de Catarata , Cooperação do Paciente , Inquéritos e Questionários , Lactente , Período Pós-Operatório , Acuidade Visual/fisiologiaRESUMO
BACKGROUND: Parents of children with cleft lip with or without cleft palate (CL/P) often face stigmatization, which has a significant impact on their quality of life and mental health. However, to date, there is a lack of comprehensive, multicenter empirical research on parents of children with CL/P in China, particularly those with large-scale samples. OBJECTIVE: This study aimed to identify major factors that contribute to the perception of stigma experienced by parents of children with CL/P. METHODS: A cross-sectional survey was conducted. A total of 104 parents of children diagnosed with CL/P in 2 hospitals were selected by convenience sampling. Demographics and disease information, the Chinese Perception of Stigma Questionnaire, the Center for Epidemiological Studies Depression Scale, and the Social Anxiety Scale were used in this study. Descriptive statistics, t tests, and one-way ANOVA were used to compare the differences between participants' demographic information and perception of stigma. Multivariable linear regression was performed to assess associations between demographic factors, social anxiety, depression, and perception of stigma. RESULTS: The mean scores for the dimensions of perception of stigma, depression, and social anxiety were 22.97 (SD 9.21), 38.34 (SD 8.25), and 22.86 (SD 6.69), respectively. Depression and social anxiety were positively associated with discrimination, while surgery status was a negatively associated variable. Parents with a college education or higher had significantly lower levels of perceived stigma compared to parents with a junior high school education (all P values <.05). These 4 factors explained 40.4% of the total model variance (F8=9.726; P<.001; R2=0.450; adjusted R2=0.404). CONCLUSIONS: Our findings highlight a concerning trend of diminished quality of life among parents of children with CL/P. Factors such as parents' education level, surgery status, depression, and social anxiety are shown to influence the level of stigma experienced. Implementing comprehensive nursing care and providing presurgical support are effective strategies for alleviating parents' social anxiety, reducing perceived stigma, and preventing depression.
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BACKGROUND: Waiting for a long time to make payments in outpatient wards and long queues of insured patients at the checkout window are common in many hospitals across China. To alleviate the problem of long queues for payment, many hospitals in China have established various mobile apps that those without health insurance can use. However, medically insured outpatients are still required to pay manually at the checkout window. Therefore, it is urgent to use information technology to innovate and optimize the outpatient service process, implement mobile payment for medically insured outpatients, and shorten the waiting time for outpatients, especially in the context of the COVID-19 epidemic. Furthermore, smartphone-based mobile payment for outpatients with health insurance could be superior to on-site cashier billing. OBJECTIVE: This study aimed to investigate the impact of smartphone-based mobile payment in relation to different aspects, such as waiting time, satisfaction with patients' waiting time, payment experience, the proportion of those dissatisfied with payment, total outpatient satisfaction, and outpatient volume, and compare mobile payment with on-site payment. METHODS: This was a historically controlled study. This study analyzed the outpatients' waiting time to make a medical insurance payment, their satisfaction with the waiting time and payment experience, the proportion of those dissatisfied with payment, and the outpatient volume of patients at Guangzhou Women and Children's Medical Center 1 year before and after the implementation of mobile payment for medical insurance in January 2021. An independent sample 2-tailed t test was used to compare waiting time, satisfaction with waiting time, and overall satisfaction. Paired sample 2-tailed t test was used to compare monthly outpatient visits. The chi-square test was used to compare the percentages of patients dissatisfied with payment. RESULTS: After the implementation of mobile payment for medical insurance outpatients, the patients' payment waiting time was significantly shortened (mean 45.28, SD 10.35 min vs mean 1.02, SD 0.25 min; t9014=53.396; P<.001), and satisfaction with waiting time and payment experience were significantly improved (mean 82.08, SD 3.17 vs mean 90.36, SD 3.45; t9014=-118.65; P<.001). Dissatisfaction with payment significantly decreased (10.27%, SD 2.18% vs 1.19% vs SD 0.30%; P<.001). The total satisfaction of outpatients significantly improved (mean 86.91, SD 3.23 vs mean 89.98, SD 3.31; t9014=-44.57; P<.001), and the outpatient volume increased (248,105.58, SD 89,280.76 vs 303,194.75, SD 53,773.12; t11=2.414; P=.03). Furthermore, payment efficiency improved, and the number of the on-site cashiers substantially decreased. CONCLUSIONS: Mobile payment for health insurance significantly shortened patients' payment waiting time; improved patient satisfaction on waiting time and payment experience and overall satisfaction; reduced the proportion of patients who were dissatisfied with payment and the cashier at the hospital; and increased monthly outpatient volume. This approach was effective and thus worthy of promoting.
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The CRISPR/Cas9 system is a versatile genome editing platform in biotechnology and therapeutics. However, the requirement of protospacer adjacent motifs (PAMs) limits the genome targeting scope. To expand this repertoire, we revisited and engineered a compact Cas9 orthologue derived from Neisseria cinerea (NcCas9) for efficient genome editing in mammal cells. We demonstrated that NcCas9 generates genome editing at target sites with N4GYAT (Y = T/C) PAM which cannot be recognized by existing Cas9s. By optimizing the NcCas9 architecture and its spacer length, editing efficacy of NcCas9 was further improved in human cells. In addition, the NcCas9-derived Base editors can efficiently generate base conversions. Six anti-CRISPR (Acr) proteins were identified as off-switches for NcCas9. Moreover, NcCas9 successfully generated efficient editing of mouse embryos by microinjection of NcCas9 mRNA and the corresponding sgRNA. Thus, the NcCas9 holds the potential to broaden the CRISPR/Cas9 toolsets for efficient gene modifications and therapeutic applications.
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Edição de Genes , Neisseria cinerea , Camundongos , Humanos , Animais , Sistemas CRISPR-Cas , Proteína 9 Associada à CRISPR/genética , Proteína 9 Associada à CRISPR/metabolismo , Neisseria cinerea/genética , Neisseria cinerea/metabolismo , Genoma , Mamíferos/genéticaRESUMO
Many therapeutic applications of CRISPR-Cas9 gene editing rely on delivery using the highly versatile adeno-associated virus (AAV) vector. The smallest type II Cas9 ortholog-Cje1Cas9, derived from Campylobacter jejuni with <1,000 amino acids-is particularly attractive for AAV delivery. However, the complex protospacer adjacent motif (PAM) of Cje1Cas9 (N3VRYAC) greatly restricts the density of recognition sequences in human genome. In this study, we identify two compact CjeCas9 orthologs designated as Cje2Cas9 and Cje3Cas9, whose PAM-interacting residues are different from those of the well-known Cje1Cas9. They can induce efficient genome editing in human cells, and their simpler trinucleotide PAM (N4CYA) requirements expand the scope of targeting. Moreover, Cje3Cas9 efficiently disrupts the Tyr gene in mice after being micro-injected into zygotes with the corresponding sgRNA. It also successfully disrupts the Pcsk9 gene in 8-week-old mouse liver after delivery with an sgRNA using an all-in-one AAV delivery vehicle. The gene-edited mice showed lower cholesterol level than wild-type mice. Notably, the 8e-nCje3-ABE and an sgRNA targeting Pcsk9 were successfully packaged into a single AAV vector for genome editing in adult mouse liver, with editing efficiency up to 12%. Thus, simple PAMs and a compact size enable Cje2/3Cas9 to expand the target scope of CRISPR-Cas9 toolsets, exhibiting considerable potential for therapeutic applications.
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Edição de Genes , Pró-Proteína Convertase 9 , Adenina , Animais , Proteína 9 Associada à CRISPR/genética , Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Camundongos , Pró-Proteína Convertase 9/genéticaRESUMO
Epigenetic alteration is a pivotal factor in tumor metastasis. PHD finger protein 13 (PHF13) is a recently identified epigenetic reader of H3K4me2/3 that functions as a transcriptional co-regulator. In this study, we demonstrate that PHF13 is required for pancreatic-cancer-cell growth and metastasis. Integrative analysis of transcriptome and epigenetic profiles provide further mechanistic insights into the epigenetic regulation of genes associated with cell metastasis during the epithelial-to-mesenchymal transition (EMT) induced by transforming growth factor ß (TGFß). Our data suggest PHF13 depletion impairs activation of TGFß stimulated genes and correlates with a loss of active epigenetic marks (H3K4me3 and H3K27ac) at these genomic regions. These observations argue for a dependency of TGFß target activation on PHF13. Furthermore, PHF13-dependent chromatin regions are enriched in broad H3K4me3 domains and super-enhancers, which control genes critical to cancer-cell migration and invasion, such as SNAI1 and SOX9. Overall, our data indicate a functional and mechanistic correlation between PHF13 and EMT.
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Proteínas de Ligação a DNA , Epigênese Genética , Transição Epitelial-Mesenquimal , Neoplasias , Fatores de Transcrição , Linhagem Celular Tumoral , Cromatina/genética , Cromatina/metabolismo , Proteínas de Ligação a DNA/genética , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/genética , Humanos , Neoplasias/patologia , Fatores de Transcrição/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
Compact CRISPR-Cas9 systems that can be packaged into an adeno-associated virus (AAV) show promise for gene therapy. However, the requirement of protospacer adjacent motifs (PAMs) restricts the target scope. To expand this repertoire, we revisited and optimized a small Cas9 ortholog derived from Streptococcus pasteurianus (SpaCas9) for efficient genome editing in vivo. We found that SpaCas9 enables potent targeting of 5'-NNGYRA-3' PAMs, which are distinct from those recognized by currently used small Cas9s; the Spa-cytosine base editor (CBE) and Spa-adenine base editor (ABE) systems efficiently generated robust C-to-T and A-to-G conversions both in vitro and in vivo. In addition, by exploiting natural variation in the PAM-interacting domain, we engineered three SpaCas9 variants to further expand the targeting scope of compact Cas9 systems. Moreover, mutant mice with efficient disruption of the Tyr gene were successfully generated by microinjection of SpaCas9 mRNA and the corresponding single guide RNA (sgRNA) into zygotes. Notably, all-in-one AAV delivery of SpaCas9 targeting the Pcsk9 gene in adult mouse liver produced efficient genome-editing events and reduced its serum cholesterol. Thus, with distinct PAMs and a small size, SpaCas9 will broaden the CRISPR-Cas9 toolsets for efficient gene modifications and therapeutic applications.
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Edição de Genes , Pró-Proteína Convertase 9 , Animais , Proteína 9 Associada à CRISPR/genética , Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas , Camundongos , Pró-Proteína Convertase 9/genética , RNA Guia de Cinetoplastídeos/genética , StreptococcusRESUMO
BACKGROUND: This study sought to develop and validate a lens opacities classification system based on ultrasound biomicroscopy (UBM) imaging to grade pediatric cataracts. METHODS: The study was conducted at Guangzhou Children's Hospital, Guangzhou Women and Children's Medical Center. UBM images of patients at the hospital from September 2013 to November 2014 were used in this study. We summarized the characteristics of lenticular opacification in each of the following 4 zones: the anterior capsule (A); the cortex (C); the nucleus (N); and the posterior capsule (P). The UBM data and intraoperative videos were compared, and sensitivity, specificity, accuracy, and positive and negative predictive values were determined for our Lens Opacities Classification System based on UBM for Pediatric Cataracts (LOCS-UP) detection. Two physicians classified pediatric cataracts (anterior capsule, cortex, and posterior capsule) by extracting 146 images from the UBM database. Patients' data were recorded to calculate the kappa coefficients. The LOCS-UP was developed. RESULTS: Under this standard, all types of pediatric cataracts can be classified and acquired a code by the LOCS-UP. The LOCS-UP had the highest sensitivity (100%) and specificity (98.96%) in naming the anterior capsule and the lowest sensitivity (50%) and specificity (89.59%) in naming the posterior capsule. Its consistency at naming the anterior capsule was satisfactory (Kappa coefficient: 0.70), and it was also able to name the nucleus, cortex, and posterior capsule (0.56, 0.58, and 0.48, respectively). CONCLUSIONS: LOCS-UP could name pediatric cataracts by providing an unique digital encoding, which could reflect characteristics exactly for different local lens anomalies to all kinds of pediatric cataract patients. This method provides detailed and accurate information about Patients' lenses.
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Ovarian cancer (OC) is the second leading cause of death in gynecological cancer. Multiple study have shown that the efficacy of tumor immunotherapy is related to tumor immune cell infiltration (ICI). However, so far, the Immune infiltration landscape of tumor microenvironment (TME) in OC has not been elucidated. In this study, We organized the transcriptome data of OC in the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases, evaluated the patient's TME information, and constructed the ICI scores to predict the clinical benefits of patients undergoing immunotherapy. Immune-related genes were further used to construct the prognostic model. After clustering analysis of ICI genes, we found that patients in ICI gene cluster C had the best prognosis, and their tumor microenvironment had the highest proportion of macrophage M1 and T cell follicular helper cells. This result was consistent with that of multivariate cox (multi-cox) analysis. The prognostic model constructed by immune-related genes had good predictive performance. By estimating Tumor mutation burden (TMB), we also found that there were multiple genes with statistically different mutation frequencies in the high and low ICI score groups. The model based on the ICI score may help to screen out patients who would benefit from immunotherapy. The immune-related genes screened may be used as biomarkers and therapeutic targets.
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Oxytocin receptor (OXTR) is involved in social behaviors, thermoregulation, and milk ejection, yet little is known about its role in breast cancer. To investigate the role of OXTR in mammary gland development and tumorigenesis, a transgenic mouse model of OXTR overexpression (++Oxtr) was used. Overexpression of OXTR-induced progressive mammary hyperplasia, unexpected milk production, and tumorigenesis in females. OXTR-induced mammary tumors showed ERBB2 upregulation and mixed histological subtypes with predomination of papillary and medullary carcinomas. OXTR overexpression led to an activation of prolactin (PRL)/p-STAT5 pathway and created a microenvironment that promotes mammary-specific tumorigenesis. PRL inhibitor bromocriptine (Br) could mitigate OXTR-driven mammary tumor growth. The study demonstrates Oxtr is an oncogene and a potential drug target for HER2-type breast cancer.
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Carcinogênese/genética , Neoplasias Mamárias Experimentais/genética , Receptores de Ocitocina/fisiologia , Animais , Transformação Celular Neoplásica/genética , Feminino , Humanos , Lactação/genética , Lactação/fisiologia , Células MCF-7 , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Animais/patologia , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Prolactina/metabolismo , Receptores de Ocitocina/genética , Fator de Transcrição STAT5/genética , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais/genética , Células Tumorais CultivadasRESUMO
BACKGROUND: The aim of the present study was to investigate the role of ultrasound biomicroscopy (UBM) in the evaluation of postoperative complications in children with congenital cataracts. METHODS: A retrospective study was conducted between September 2012 and December 2016 at Guangzhou Women and Children's Medical Center. Red reflex test and high-resolution bag/balloon UBM were performed to evaluate postoperative congenital cataracts. The red reflex test results were recorded, and UBM imaging results were recorded and analyzed. Different postoperative complications were classified based on the UBM imaging features, and a second procedure was performed accordingly. The UBM images were compared with the images captured from the intraoperative videos. RESULTS: In total, we looked at 120 eyes in 96 patients (65 males and 31 females) in the present study. The age of the cohort was 3-76 months. A total of 51 eyes with poor red reflex were included. There were complications in 46 eyes after congenital cataract surgery, as detected by UBM, including posterior capsular opacification (n=29 eyes), pupil block (n=8 eyes), synechia (n=5 eyes), hyphema (n=1 eye), and abnormal intraocular lens (IOL) placement (n=3 eyes). UBM images showed specific features of postoperative complications. CONCLUSIONS: UBM is a valuable tool for the early evaluation of postoperative complications of congenital cataracts, especially for those with media opacities or when pupil dilation is not possible.
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Dedifferentiation of chondrocyte greatly restricts its function and application, however, it is poorly understood except a small number of canonical markers. The non-cell-adhesive property endows polysaccharide hydrogel with the ability to maintain chondrocyte phenotype, which can serve as a platform to identify new molecular markers and therapeutic targets of chondrocyte dedifferentiation. In this study, the high-throughput RNA sequencing (RNA-seq) was first performed on articular chondrocytes at primary (P0) and passage 1 (P1) stages to explore the global alteration of gene expression along with chondrocyte dedifferentiation. Significantly, several potential marker genes, such as PFKFB3, KDM6B, had been identified via comparatively analyzing their expression in P0 and P1 chondrocytes as well as in 3D constructs (i.e. chondrocyte-laden alginate hydrogel and HA-MA hydrogel) at both mRNA and protein level. Besides, the changes in cellular morphology and enriched pathway of differentially expressed genes during chondrocyte dedifferentiation was studied in detail. This study developed the use of hydrogel as a platform to investigate chondrocyte dedifferentiation; the results provided new molecular markers and potential therapeutic targets of chondrocyte dedifferentiation.
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ABSTRACT: Early enteral nutrition (EN) promotes the recovery of critically ill patients, but the initiation time for EN in neonates after cardiac surgery remains unclear.This study aimed to investigate the effect of initiation time of EN after cardiac surgery in neonates with complex congenital heart disease (CHD).Neonates with complex CHD admitted to the CICU from January 2015 to December 2017 were retrospectively analyzed. Patients were divided into the 24-hour Group (initiated at 24âhours after surgery in 2015) (nâ=â32) and 6-hour Group (initiated at 6âhours after surgery in 2016 and 2017) (nâ=â66). Data on the postoperative feeding intolerance, nutrition-related laboratory tests (albumin, prealbumin, retinol binding protein), and clinical outcomes (including duration of mechanical ventilation, CICU stay, and postoperative hospital stay) were collected.The incidence of feeding intolerance was 56.3% in 24-hour Group and 39.4%, respectively (Pâ=â.116). As compared to 24-hour Group, prealbumin and retinol binding protein levels were higher (160.7â±â64.3 vs 135.2â±â28.9âmg/L, Pâ=â.043 for prealbumin; 30.7â±â17.7 vs 23.0â±â14.1âg/L Pâ=â.054 for retinol-binding protein). The duration of CICU stay (9.4â±â4.5 vs 13.3â±â10.4 day, Pâ=â.049) and hospital stay (11.6â±â3.0 vs 15.8â±â10.3 day, Pâ=â.028) were shorter in 6-hour Group.Early EN improves nutritional status and clinical outcomes in neonates with complex CHD undergoing cardiac surgery, without significant feeding intolerance.
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Nutrição Enteral/métodos , Cardiopatias Congênitas/cirurgia , Fatores de Tempo , Distribuição de Qui-Quadrado , Unidades de Cuidados Coronarianos/organização & administração , Unidades de Cuidados Coronarianos/estatística & dados numéricos , Nutrição Enteral/normas , Nutrição Enteral/estatística & dados numéricos , Feminino , Cardiopatias Congênitas/dietoterapia , Hospitalização/estatística & dados numéricos , Humanos , Recém-Nascido , Masculino , Estudos Prospectivos , Estudos RetrospectivosRESUMO
Retinopathy of prematurity (ROP) has become the main cause of blindness in children. To study resilience, anxiety and depression, coping style and social support and their correlation in parents of premature infants undergoing outpatient fundus examination for ROP, and thereby provide evidence for clinical intervention. Questionnaire surveys were conducted by using General Information Questionnaire, Connor-Davidson Resilience Scale, Self-rating Anxiety Scale, Self-rating Depression Scale, Social Support Scale, and the Simplified Coping Style Questionnaire. The resilience in the parents scored (63.24 ± 12.97) points, which was lower than that of Chinese norm (t = 2.309, P < 0.05). The scores of anxiety and depression were higher than those of Chinese norm (t = 12.592, t = 2.362, both P < 0.05). The score of social support was lower than that of Chinese norm (t = 3.793, p < 0.01). Resilience was negatively correlated with anxiety and depression (r = -0.287, -0.358, both P < 0.01), and positively correlated with coping tendency and social support (r = 0.299, 0.139, both P < 0.05). These findings suggested that the parents have low resilience, high levels of anxiety and depression, and low level of social support. Medical staff should assess and improve their psychological status.
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Pais , Retinopatia da Prematuridade , Adaptação Psicológica , Assistência Ambulatorial , Ansiedade/epidemiologia , China/epidemiologia , Depressão/epidemiologia , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Pais/psicologia , Exame Físico , Resiliência Psicológica , Retinopatia da Prematuridade/diagnóstico , Fatores de Risco , Apoio Social , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Procedural pain is underestimated in hospitalized preterm infants. The aim of this study was to assess the reliability, validity, and clinical utility of the Neonatal Facial Coding System (NFCS), Douleur Aiguë du Nouveau-né (DAN) scale, Neonatal Infant Pain Scale (NIPS), and Premature Infant Pain Profile (PIPP) in premature infants undergoing heel blood collection. We assume that the four scales were similar in reliablility and validity (but different in clinical utility). METHODS: The pain assessments were performed on 111 premature infants using the four scales. Internal consistency was determined by Cronbach's α, and the reliability was determined by the intraclass correlation coefficients. Concurrent validity was evaluated by Spearman's rank correlations. Bland-Altman plots were used to investigate the convergent validity. RESULTS: The internal consistency and their reliability of the scales were high (p < 0.001). Scores were significantly higher at the time of blood collection (p < 0.001). Mean scores of clinical utility of PIPP were significantly higher than NFCS and DAN (p < 0.05) but not higher than the NIPS (p > 0.05). CONCLUSIONS: The four scales were reliable and valid. This study suggests that the PIPP and NIPS has good clinical utility and are better choice for evaluating procedural pain in premature infants. IMPACT: The aim of this study was to assess the reliability, validity, and clinical utility of NFCS, DAN, NIPS, and PIPP in premature infants undergoing heel blood collection. The results showed that the four scales have high reliability and internal consistency; the PIPP and NIPS have good clinical utility and are better choice for evaluating procedural pain in premature infants. Our study results provided a reference for clinical workers in choosing pain assessment scales and conduction intervention.
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Calcanhar/irrigação sanguínea , Medição da Dor/métodos , Dor Processual/diagnóstico , Manejo de Espécimes/efeitos adversos , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Reprodutibilidade dos TestesRESUMO
The role of histone ubiquitination in directing cell lineage specification is only poorly understood. Our previous work indicated a role of the histone 2B ubiquitin ligase RNF40 in controlling osteoblast differentiation in vitro. Here, we demonstrate that RNF40 has a stage-dependent function in controlling osteoblast differentiation in vivo. RNF40 expression is essential for early stages of lineage specification, but is dispensable in mature osteoblasts. Paradoxically, while osteoblast-specific RNF40 deletion led to impaired bone formation, it also resulted in increased bone mass due to impaired bone cell crosstalk. Loss of RNF40 resulted in decreased osteoclast number and function through modulation of RANKL expression in OBs. Mechanistically, we demonstrate that Tnfsf11 (encoding RANKL) is an important target gene of H2B monoubiquitination. These data reveal an important role of RNF40-mediated H2B monoubiquitination in bone formation and remodeling and provide a basis for exploring this pathway for the treatment of conditions such as osteoporosis or cancer-associated osteolysis.
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Histonas/metabolismo , Osteócitos/metabolismo , Ligante RANK/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Diferenciação Celular , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Osteogênese/fisiologia , Ligante RANK/genética , Ubiquitina-Proteína Ligases/genética , Ubiquitinação/fisiologiaRESUMO
CRISPR-Cas9-mediated gene knockout and base-editing-associated induction of STOP codons (iSTOP) have been widely used to exterminate the function of a coding gene, while they have been reported to exhibit side effects. In this study, we propose a novel and practical alternative method referred to as CRISPR Start-Loss (CRISPR-SL), which eliminates gene expression by utilizing both adenine base editors (ABEs) and cytidine base editors (CBEs) to disrupt the initiation codon (ATG). CRISPR-SL has been verified to be a feasible strategy on the cellular and embryonic levels (mean editing efficiencies up to 30.67% and 73.50%, respectively) and in two rabbit models mimicking Otc deficiency (Otc gene) and long hair economic traits (Fgf5 gene).
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Direct reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) requires a resetting of the epigenome in order to facilitate a cell fate transition. Previous studies have shown that epigenetic modifying enzymes play a central role in controlling induced pluripotency and the generation of iPSC. Here we show that RNF40, a histone H2B lysine 120 E3 ubiquitin-protein ligase, is specifically required for early reprogramming during induced pluripotency. Loss of RNF40-mediated H2B monoubiquitination (H2Bub1) impaired early gene activation in reprogramming. We further show that RNF40 contributes to tissue-specific gene suppression via indirect effects by controlling the expression of the polycomb repressive complex-2 histone methyltransferase component EZH2, as well as through more direct effects by promoting the resolution of H3K4me3/H3K27me3 bivalency on H2Bub1-occupied pluripotency genes. Thus, we identify RNF40 as a central epigenetic mediator of cell state transition with distinct functions in resetting somatic cell state to pluripotency.
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Células-Tronco Pluripotentes Induzidas/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Diferenciação Celular/fisiologia , Reprogramação Celular/fisiologia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Processamento de Proteína Pós-Traducional , Ubiquitina-Proteína Ligases/genéticaRESUMO
BACKGROUND: In China, a long waiting time for registration is a common occurrence in many tertiary hospitals. This study aimed to analyze the effects of a comprehensive reservation service for non-emergency registration on appointment registration rate, patient waiting time, patient satisfaction and outpatient volume at the Guangzhou Women and Children's Medical Center. METHODS: This was a cross-sectional study. This study investigated the effects of a comprehensive reservation service for non-emergency registration in Guangzhou Women and Children's Medical Center in China starting in October 2015. In total, 2194 patients completed a satisfaction survey administered by the Guangdong Situation Research Center. The content of the questionnaire consisted of six aspects: general impression, service attitude, service quality, hospital environment, price perception and medical ethics. A Likert 5-point rating scale was used in the questionnaire; answers were classified as "very satisfied", "relatively satisfied", "neutral", "unsatisfied" and "very unsatisfied". The method of application was paper-based. T-tests were used to compare the sample means, and chi-square tests were used to compare the rates. A multiple-test procedure was performed to evaluate the differences in the reservation rates during a 12-month period. RESULTS: After the implementation of the comprehensive reservation service for non-emergency registration in our hospital, which has an annual outpatient volume of approximately 4 million, the monthly appointment registration rate increased from (34.95 ± 2.91)% to(89.13 ± 3.12)%,P < 0.01. The patient waiting time was significantly reduced (P < 0.01), and the proportion of patients who believed that the waiting time required improvement was decreased significantly (P < 0.01). Moreover, the third-party evaluation result of outpatient satisfaction significantly improved (P < 0.01). The total hospital outpatient volume decreased(P < 0.01). The outpatient volume of the Department of General Pediatrics decreased. CONCLUSION: The implementation of the comprehensive reservation service for non-emergency registration in the hospital shortened patient waiting time and improved patient satisfaction, and the outpatient volume was effectively controlled. These results indicated that this program obtained the desired results in a Grade 3A hospital in China.
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Agendamento de Consultas , Pacientes Ambulatoriais/estatística & dados numéricos , Satisfação do Paciente/estatística & dados numéricos , Centros de Atenção Terciária/organização & administração , Listas de Espera , China , Estudos Transversais , Feminino , Pesquisa sobre Serviços de Saúde , Humanos , Masculino , Inquéritos e Questionários , Fatores de TempoRESUMO
BACKGROUND: In many clinics, patients now have the option to make Web-based appointments but doing so according to their own judgment may lead to wrong registration and delayed medical services. We hypothesized that smartphone-based triage in outpatient services is superior to Web-based self-appointment registration guided by the medical staff. OBJECTIVE: This study aimed to investigate smartphone-based triage in outpatient services compared with Web-based self-appointment registration and to provide a reference for improving outpatient care under appointment registration. METHODS: The following parameters in Guangzhou Women and Children's Medical Center were analyzed: wrong registration rate, the degree of patient satisfaction, outpatient visits 6 months before and after smartphone-based triage, queries after smartphone-based triage, number of successful registrations, inquiry content, and top 10 recommended diseases and top 10 recommended departments after queries. RESULTS: Smartphone-based triage showed significant effects on average daily queries, which accounted for 16.15% (1956/12,112) to 29.46% (3643/12,366) of daily outpatient visits. The average daily successful registration after queries accounted for 56.14% (1101/1961) to 60.92% (1437/2359) of daily queries and 9.33% (1130/12,112) to 16.83% (2081/12,366) of daily outpatient visits. The wrong registration rate after smartphone-based triage was reduced from 0.68% (12,810/1,895,829) to 0.12% (2379/2,017,921) (P<.001), and the degree of patient satisfaction was improved. Monthly outpatient visits were increased by 0.98% (3192/325,710) to 13.09% (42,939/328,032) compared with the same period the preceding year (P=.02). CONCLUSIONS: Smartphone-based triage significantly reduces the wrong registration rate caused by patient Web-based appointment registration and improves the degree of patient satisfaction. Thus, it is worth promoting.