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AIM: Natural orifice specimen extraction (NOSE) is an alternative to conventional transabdominal retrieval. We aimed to compare outcomes following transvaginal specimen extraction (TVSE) and transabdominal specimen extraction (TASE) in minimally invasive abdominal surgery. METHODS: An electronic database search of PubMed, Embase and CENTRAL was performed from inception until March 2023. Comparative studies evaluating TVSE versus TASE in adult female patients were included. Studies involving transanal NOSE, endoluminal surgery, or TVSE with concomitant hysterectomy were excluded. Weighted mean differences (WMD) and odds ratio were estimated for continuous and dichotomous outcomes respectively. Primary outcomes were postoperative day 1 (POD1) pain and length of stay (LOS). Secondary outcomes were operative time, rescue analgesia, morbidity, and cosmesis. A review of sexual, oncological, and technical outcomes was performed. RESULTS: Thirteen studies (2 randomised trials, 11 retrospective cohort studies), involving 1094 patients (TASE 583, TVSE 511), were included in the analysis. Seven studies involved colorectal disease and six assessed gynaecological conditions. TVSE resulted in significantly decreased POD1 pain (WMD 1.08, 95% CI: 0.49, 1.68) and shorter LOS (WMD 1.18 days, 95% CI: 0.14, 2.22), compared to TASE. Operative time was similar between both groups, with fewer patients requiring postoperative rescue analgesia with TVSE. Overall morbidity rates, as well as both wound-related and non-wound related complication rates were better with TVSE, while anastomotic morbidity rates were comparable. Cosmetic scores were higher with TVSE. TVSE did not result in worse sexual or oncological outcomes. CONCLUSION: TVSE may be feasible and beneficial compared to TASE when performed by proficient laparoscopic operators, using appropriate selection criteria. Continued evaluation with prospective studies is warranted.
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Procedimentos Cirúrgicos Minimamente Invasivos , Vagina , Humanos , Feminino , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Vagina/cirurgia , Cirurgia Endoscópica por Orifício Natural/métodos , Cirurgia Endoscópica por Orifício Natural/efeitos adversos , Tempo de Internação , Duração da CirurgiaRESUMO
PURPOSE: Triple-negative breast cancer (TNBC) represents the worst prognostic subtype of breast cancer and lacks targeted therapeutic drugs. Signal transducer and activator of transcription 3 (STAT3) is overexpressed and constitutively activated in TNBCs and associated with poor patient outcomes. However, no agents targeting STAT3 have been successfully developed and marketed. Selective Estrogen Receptor Modulators (SERMs) have been reported as potential inhibitors of the IL-6/STAT3 signaling pathway. Naphthalene compounds have good pharmacological activity and significant anti-cancer activity. In this study, we synthesized a new series of naphthalene derivatives with the general structure of SERM and evaluated their effects on TNBC and STAT3 signals. METHODS: A new series of compounds based on the scaffold of SERMs and an amino group were designed and screened based on the structure-activity relationship by MTT assay. The binding activity of SMY002 to STAT3 was predicted and validated by docking and SPR. The STAT3 signaling target and anti-cancer effects of SMY002 were evaluated with three TNBC cell lines and the mice transplanted tumor model. RESULTS: Among the compounds, SMY002 displayed the most potent activity, which could directly interact with STAT3 SH2-domain, and strongly inhibit the phosphorylation, dimerization, nuclear distribution, transcriptional activity, and target genes expression of STAT3. Furthermore, SMY002 markedly suppressed migration, invasion, survival, growth, and metastasis of TNBC cells in vitro and in vivo via down-regulating the expression of Cyclin D1 and MMP9. CONCLUSIONS: SMY002 can significantly inhibit the growth and metastasis of TNBC cells by targeting the STAT3 signal.
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Neoplasias de Mama Triplo Negativas , Humanos , Animais , Camundongos , Neoplasias de Mama Triplo Negativas/patologia , Fator de Transcrição STAT3/metabolismo , Moduladores Seletivos de Receptor Estrogênico/farmacologia , Transdução de Sinais , Proliferação de Células , Naftalenos/farmacologia , Naftalenos/uso terapêutico , Linhagem Celular TumoralRESUMO
We describe a straightforward self-assembly route to nanoporous materials derived from a hexagonally-packed cylinder (HEX) morphology of a polyisoprene-block-polylactide (PI-b-PLA) diblock copolymer, by thermal cross-linking of the minority PI domains followed by selective chemical etching of the PLA matrix. The resulting mechanically stable and porous samples defy the expectation that the remaining cylinders cannot yield a robust, integrated material upon matrix removal. Scanning electron microscopy imaging reveals that this unexpected structural integrity stems from the interconnected nanofibrils therein, reflecting topological defects at the grain boundaries of the parent polydomain HEX nanostructure. Hydrodynamic radius-dependent poly(ethylene oxide) (Mn = 0.4-35 kg/mol) permeation behavior through these monoliths directly demonstrated the continuity and size selectivity of the nanoporous material. The ready accessibility of block copolymer HEX morphologies of varied chemistries suggests that this matrix etching strategy will enable the future design of functional, size-selective nanofiltration membrane materials.
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Chromatin immunoprecipitation (ChIP) is the gold-standard method for detection of interactions between proteins and chromatin and is a powerful tool for identification of epigenetic modifications. Although ChIP protocols for plant species have been developed, many specific features of plants, especially woody plants, still hinder the efficiency of immunoprecipitation, resulting in inefficient ChIP enrichment and an active demand for a highly efficient ChIP protocol. In this study, using birch (Betula platyphylla) and Arabidopsis (Arabidopsis thaliana) as the research materials, we identified five factors closely associated with ChIP efficiency, including crosslinking, concentration of chromatin using centrifugal filters, use of a different immunoprecipitation buffer, rescue of DNA with proteinase K, and use of Suc to increase immunoprecipitation efficiency. Optimization of any these factors can significantly improve ChIP efficiency. Considering these factors together, we developed a robust ChIP protocol that achieved a 14-fold improvement in ChIP enrichment for birch and a >6-fold improvement for Arabidopsis compared to the standard ChIP method. As this ChIP method works well in both birch and Arabidopsis, it should also be suitable for other woody and herbaceous species. In addition, this ChIP method enables detection of low-abundance transcription factor-DNA interactions and may extend the application of ChIP in the plant kingdom.
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Arabidopsis/genética , Arabidopsis/metabolismo , Betula/genética , Betula/metabolismo , Cromatina/genética , Cromatina/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Imunoprecipitação da Cromatina/métodos , Epigênese Genética , Regulação da Expressão Gênica de Plantas , Genes de PlantasRESUMO
BACKGROUND: In recent years, there have been numerous studies exploring different teaching methods for improving diagnostic reasoning in undergraduate medical students. This systematic review examines and summarizes the evidence for the effectiveness of these teaching methods during clinical training. METHODS: PubMed, Embase, Scopus, and ERIC were searched. The inclusion criteria for the review consist of articles describing (1) methods to enhance diagnostic reasoning, (2) in a clinical setting (3) on medical students. Articles describing original research using qualitative, quantitative, or mixed study designs and published within the last 10 years (1 April 2009-2019) were included. Results were screened and evaluated for eligibility. Relevant data were then extracted from the studies that met the inclusion criteria. RESULTS: Sixty-seven full-text articles were first identified. Seventeen articles were included in this review. There were 13 randomized controlled studies and four quasi-experimental studies. Of the randomized controlled studies, six discussed structured reflection, four self-explanation, and three prompts for generating differential diagnoses. Of the remaining four studies, two employed the SNAPPS1 technique for case presentation. Two other studies explored schema-based instruction and using illness scripts. Twelve out of 17 studies reported improvement in clinical reasoning after the intervention. All studies ranked level two on the New World Kirkpatrick model. DISCUSSION: The authors posit a framework to teach diagnostic reasoning in the clinical setting. The framework targets specific deficiencies in the students' reasoning process. There remains a lack of studies comparing the effectiveness of different methods. More comparative studies with standardized assessment and evaluation of long-term effectiveness of these methods are recommended.
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Educação de Graduação em Medicina , Estudantes de Medicina , Competência Clínica , Humanos , Resolução de ProblemasRESUMO
Punch-sticking during tablet compression is a common problem for many active pharmaceutical ingredients (APIs), which renders tablet formulation development challenging. Herein, we demonstrate that the punch-sticking propensity of a highly sticky API, celecoxib (CEL), can be effectively reduced by spherical crystallization enabled by a polymer assisted quasi-emulsion solvent diffusion (QESD) process. Among three commonly used pharmaceutical polymers, poly(vinylpyrrolidone) (PVP), hydroxypropyl cellulose (HPC), and hydroxypropyl methylcellulose (HPMC), HPMC was the most effective in stabilizing the transient emulsion during QESD and retarding the coalescence of emulsion droplets and the initiation of CEL crystallization. These observations may arise from stronger intermolecular interactions between HPMC and CEL, consistent with solution 1H NMR analyses. SEM and X-ray photoelectron spectroscopy confirmed the presence of a thin layer of HPMC on the surfaces of spherical particles. Thus, the sticking propensity was significantly reduced because the HPMC coating prevents direct contact between CEL and the punch tip during tablet compression.
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Celecoxib/química , Emulsões/química , Polímeros/química , Solventes/química , Cristalização/métodos , Difusão , Composição de Medicamentos/métodos , Derivados da Hipromelose/química , Comprimidos/químicaRESUMO
Trihelix transcription factors are characterized by containing a conserved trihelix (helix-loop-helix-loop-helix) domain that binds to GT elements required for light response, and they play roles in light stress and in abiotic stress responses. However, only a few of them have been functionally characterized. In the present study, we characterized the function of AST1 (Arabidopsis SIP1 clade Trihelix1) in response to salt and osmotic stress. AST1 shows transcriptional activation activity, and its expression is induced by osmotic and salt stress. A conserved sequence highly present in the promoters of genes regulated by AST1 was identified, which was bound by AST1, and termed the AGAG-box with the sequence [A/G][G/A][A/T]GAGAG. Additionally, AST1 also binds to some GT motifs including the sequence of GGTAATT, TACAGT, GGTAAAT and GGTAAA, but failed in binding to the sequence of GTTAC and GGTTAA. Chromatin immunoprecipitation combined with quantitative real-time reverse transcription-PCR analysis suggested that AST1 binds to the AGAG-box and/or some GT motifs to regulate the expression of stress tolerance genes, resulting in reduced reactive oxygen species, Na+ accumulation, stomatal apertures, lipid peroxidation, cell death and water loss rate, and increased proline content and reactive oxygen species scavenging capability. These physiological changes affected by AST1 finally improve salt and osmotic tolerance.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Motivos de Nucleotídeos/genética , Osmose , Tolerância ao Sal , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Sequência de Bases , Morte Celular , Dessecação , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Genes de Plantas , Genótipo , Malondialdeído/metabolismo , Estômatos de Plantas/fisiologia , Plantas Geneticamente Modificadas , Potássio/metabolismo , Prolina/metabolismo , Ligação Proteica , Transporte Proteico , Espécies Reativas de Oxigênio/metabolismo , Reprodutibilidade dos Testes , Tolerância ao Sal/genética , Análise de Sequência de RNA , Sódio/metabolismo , Frações Subcelulares/metabolismo , Fatores de Tempo , Fatores de Transcrição/genéticaRESUMO
Interconnected macroporous polymers were prepared by copolymerizing methyl acrylate (MA) via Pickering high internal phase emulsion (HIPE) templates with modified silica particles. The pore structure of the obtained polymer foams was observed by field-emission scanning electron microscopy (FE-SEM). Gas permeability was characterized to evaluate the interconnectivity of macroporous polymers. The polymerization shrinkage of continuous phase tends to form open pores while the solid particles surrounding the droplets act as barriers to produce closed pores. These two conflicting factors are crucial in determining the interconnectivity of macroporous polymers. Thus, poly-Pickering HIPEs with high permeability and well-defined pore structure can be achieved by tuning the MA content, the internal phase fraction, and the content of modified silica particles.
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Mild traumatic brain injury (MTBI) is defined as a mild brain trauma resulting in a short loss of consciousness and alteration of mental status. It may also occasionally develop persistent and progressive symptoms. It has been confirmed that MTBI causes changes of anatomic structures in central nervous system and biomarkers in the body fluid. However, there is no sufficient research on relevance among threshold for the brain injury, individual vulnerability and duration of disturbance of consciousness. Furthermore, there are no reliable diagnostic methods to establish whether a blow to the head is sufficient to cause the brain injury. This review provides references for biomarkers in cerebrospinal fluid and blood associated with TBI. It also provides application status and potential prospects for further assessment and diagnosis of MTBI.
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Biomarcadores/sangue , Biomarcadores/líquido cefalorraquidiano , Lesões Encefálicas/sangue , Lesões Encefálicas/líquido cefalorraquidiano , Concussão Encefálica/complicações , Lesões Encefálicas/diagnóstico , Lesões Encefálicas/etiologia , Progressão da Doença , HumanosRESUMO
BACKGROUND: Nexrutine is an herbal extract derived from Phellodendron amurense, known for its anti-inflammatory, antidiarrheal, and hemostatic properties. However, its effect on ulcerative colitis (UC) remains unclear. METHODS: A mouse model of UC was induced by 3% dextran sulfate sodium, while human colonic epithelial cells NCM-460 were exposed to lipopolysaccharide. Both models were treated with Nexrutine at 300 or 600 mg/kg, with Mesalazine applied as a positive control regimen. The disease activity index (DAI) of mice was calculated, and the pathological injury scores were assessed through hematoxylin and eosin staining. The viability of NCM-460 cells was determined using the CCK-8 method. Inflammatory cytokines were detected using ELISA kits. Expression of mucin 3 (MUC3), Claudin-1, and tight junction protein (ZO-1) was detected to analyze mucosal barrier integrity. Target genes of Nexrutine were predicted using bioinformatics tools. Expression of RELA proto-oncogene (RELA) was analyzed using qPCR and western blot assays. RESULTS: The Nexrutine treatments significantly alleviated DAI of mice, mitigated pathological changes in their colon tissues, decreased the production of pro-inflammatory cytokines, enhanced the barrier integrity-related proteins, and increased NCM-460 cell viability in vitro. RELA, identified as a target gene of Nexrutine, showed elevated levels in UC models but was substantially suppressed by Nexrutine treatment. Adenovirus-mediated RELA upregulation in mice or the overexpression plasmid of RELA in cells counteracted the effects of Nexrutine treatments, exacerbating UC-related symptoms. CONCLUSION: This study demonstrates that Nexrutine alleviates inflammatory mucosal barrier damage in UC by suppressing RELA transcription.
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Colite Ulcerativa , Extratos Vegetais , Humanos , Animais , Camundongos , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/tratamento farmacológico , Mesalamina , Citocinas , Inflamação/tratamento farmacológico , Fator de Transcrição RelA/genéticaRESUMO
Drought stress poses a serious threat to Pinus massoniana seedling growth in southern China. Trichoderma species, as beneficial microorganisms, have been widely used in agriculture to enhance plant growth and drought tolerance, but the interaction mechanisms remain unclear. To investigate the effect of drought-resistant Trichoderma longibrachiatum inoculation on P. massoniana growth under drought stress, the plant physiological indicators and rhizosphere microbiome diversity were measured to identify Trichoderma-activated mechanisms. Trichoderma longibrachiatum inoculation significantly promoted P. massoniana growth under drought treatment, and enhanced nitrogen, phosphorus, and potassium absorption compared with those of non-inoculated seedlings. Trichoderma longibrachiatum treatment alleviated the damage to cell membranes and needle tissue structure, and significantly increased antioxidant enzyme activities, osmotic substance contents, and photosynthesis in P. massoniana in response to drought stress. Soil nutrient contents, activities of sucrase, phosphatase, and urease as well as the relative abundances of the dominant genera Burkholderia, Rhodanobacter, and Trichoderma were elevated in the rhizosphere soil of P. massoniana inoculated with T. longibrachiatum under drought stress. A network analysis showed that certain crucial dominant taxa driven by T. longibrachiatum inoculation, including Penicillium, Trichoderma, Simplicillium, Saitozyma, Burkholderia, Bradyrhizobium, Sinomonas, and Mycobacterium, had more correlations with other microorganisms in the soil. Trichoderma longibrachiatum enhanced P. massoniana seedling growth under drought stress by regulating physiological responses and soil microbial community.
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BACKGROUND: Circulating cytokines have been associated with colorectal cancer (CRC). However, their causal correlation remains undetermined. This investigation uses genetic data to evaluate the mechanism that links circulating cytokines and CRC via Mendelian Randomization (MR). METHODS: A two-sample MR evaluation was carried out to investigate the mechanism associating circulating cytokines and CRC in individuals of European ancestry. The Genome-wide association studies statistics, which are publically accessible, were used. Eligible instrumental SNPs that were significantly related to the circulating cytokines were selected. Multiple MR analysis approaches were carried out, including Simple Mode, inverse variance weighted (IVW), MR-Egger, Weighted Mode, Weighted Median, and MR pleiotropy residual sum and outlier (MR-PRESSO) methods. RESULTS: The evidence supporting the association of genetically predicted circulating levels with the increased risk of CRC was revealed; these included vascular endothelial growth factor (OR = 1.352, 95% CI: 1.019-1.315, P = 0.024), interleukin-12p70 (OR = 1.273, 95% CI: 1.133-1.430, P = 4.68×10-5), interleukin-13 (OR = 1.149, 95% CI: 1.012-1.299, P = 0.028), interleukin-10 (OR = 1.230, 95% CI: 1.013-1.493, P = 0.037), and interleukin-7 (OR = 1.191, 95% CI: 1.023-1.386 P = 0.024). Additionally, MR analysis negative causal association between macrophage colony stimulating factor and CRC (OR = 0.854, 95% CI: 0.764-0.955, P = 0.005). The data from Simple Mode, Weighted Median, MR-Egger, and Weighted Mode analyses were consistent with the IVW estimates. Furthermore, the sensitivity analysis indicated that the presence of no horizontal pleiotropy to bias the causal estimates. CONCLUSION: This investigation identified a causal association between circulating cytokines levels risk of CRC and may provide a deeper understanding of the pathogenesis of CRC, as well as offer promising leads for the development of novel therapeutic targets for CRC.
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Neoplasias Colorretais , Citocinas , Humanos , Citocinas/genética , Estudo de Associação Genômica Ampla , Análise da Randomização Mendeliana , Fator A de Crescimento do Endotélio Vascular , Neoplasias Colorretais/genéticaRESUMO
Klebsiella pneumoniae, a notorious pathogen for opportunistic health care-associated infections, represents increasing multidrug resistance, particularly to carbapenems. OXA-232 carbapenemase, as a variant of OXA-48, has been increasingly reported worldwide. ST231, an epidemic, multidrug resistant (MDR) K. pneumoniae clone in south and southeast Asia, has been found in other regions, including Europe. In the study, five OXA-232 carbapenemase-producing Klebsiella pneumoniae isolates, four of which belong to sequence type 231 (ST231) and one of which belongs to ST15, were isolated from two hospitals in China. All isolates displayed a MDR phenotype, being susceptible to only polymyxin B and colistin, and the blaOXA-232 gene was located on a ColKP3-type nonconjugative plasmid of 6.1 kb. A phylogenetic analysis of the global ST231 K. pneumoniae isolates (n = 231) suggested that the four ST231 isolates from this study gathered with strains from south Asia (especially India), indicating that the emerging Chinese ST231 clone was more closely related to south Asia isolates and might have spread from south Asia, where ST231 was a successful epidemic clone. Virulence assays suggested that the four ST231 strains were not highly virulent, as they displayed significantly lower virulence potential, compared with a ST23 K1 hypervirulent isolate in a G. mellonella infection and in mouse intraperitoneal infection models, although three ST231 strains harbored a plasmid-borne aerobactin-encoding iuc gene cluster. This is the first report of ST231 K. pneumoniae clinical strains bearing blaOXA-232 in China, and it highlights the emergence of the ST231 clone causing bloodstream infections in a health care setting as well as calls attention to the transmission of this emerging clone in China. IMPORTANCE OXA-232 carbapenemase, being a vital resistance mechanism against carbapenems, has recently been increasingly reported. In China, the identified OXA-232-producing K. pneumoniae isolates almost belonged to ST15 and were not hypervirulent, despite harboring a virulence plasmid. Here, we report the first occurrence in China of a MDR OXA-232-producing K. pneumoniae ST231 clone that is an epidemic ST type in south and southeast Asia. A phylogenetic analysis indicated that this emerging Chinese ST231 clone was more closely related to Indian isolates. The occurrence of this clone may have been driven through the transnational importation of Indian ST231 K. pneumoniae clones. Moreover, this study is the first to assess the virulence potential of ST231 clones that have never been estimated in previous studies. While the high burden of MDR K. pneumoniae is concerning, genomic surveillance can shed light on the transmission chains of novel MDR clones, and active surveillance should be enforced to restrict the spread of MDR isolates.
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We conducted a meta-analysis of IOTA (international ovarian tumor analysis) ADNEX (Assessment of Different NEoplasias in the adneXa) as ultrasound system and MRI (magnetic resonance imaging) ADNEX scoring systems as MR system to assess their diagnostic test accuracy for differentiating benign from malignant adnexal masses of the ovary. We performed an electronic search for relevant publications in the English language up to February 2021 using PubMed, CENTRAL (Cochrane Central Register of Controlled Trials), Web of Science, and Google scholar databases and search engines. We computed the pooled sensitivity, pooled specificity, and summary receiver operating characteristics curve (SROC) using the statistical software STATA (Version 13, College Station, TX, StataCorp LP). Based on 11 studies using IOTA-ADNEX, we observed pooled sensitivity, specificity, area under curve, and diagnostic odds ratio were 96% (95% CI, 94% to 97%), 79% (95% CI, 70% to 86 %), 97% (95% CI, 95% to 98%), and 88 (95% CI, 43 to 180). Based on five studies using MR-ADNEX scoring system the pooled sensitivity, specificity, area under curve and diagnostic odds ratio were 91 % (95% CI, 87% to 94 %), 95% (95% CI, 92% to 97 %), 98% (95% CI, 96% to 99%), and 189 (95% CI, 90 to 396) respectively. Our meta-analysis results demonstrate that the MR-ADNEX scoring system had higher specificity however bit lower sensitivity compared to the IOTA-ADNEX scoring system for discriminating benign from malignant ovarian tumors.
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Doenças dos Anexos , Neoplasias Ovarianas , Anexos Uterinos/diagnóstico por imagem , Anexos Uterinos/patologia , Doenças dos Anexos/diagnóstico por imagem , Doenças dos Anexos/patologia , Diagnóstico Diferencial , Feminino , Humanos , Imageamento por Ressonância Magnética , Neoplasias Ovarianas/diagnóstico por imagem , Sensibilidade e Especificidade , Ultrassonografia/métodosRESUMO
Catheter-associated urinary tract infection (CAUTI) is the most common complication in patients with indwelling catheterization. The incidence of CAUTI in my country is still at a relatively high level compared with foreign countries, especially for the ICU, which has a high usage rate of urinary catheters, to focus on prevention and control. This article focuses on studying the risk factors of CAUTI in critically ill patients and discusses targeted preventive care measures. This article investigates and examines the clinical data of CAUTI in critically ill patients. After statistical analysis, the risk factors that affect CAUTI are summarized, so as to derive the cause of CAUTI in order to strengthen clinical care and to further study the prevention, control, and nursing of CAUTI to provide reference. Clinical data shows that the CAUTI infection rate of patients with catheter indwelling ≥7 days is greater than that of patients with catheter indwelling days less than 7 days. The CAUTI infection rate of the patients who change the urine collection bag every day or ≥7 days is greater than that of the patients who change the urine collection bag within 2 to 4 days.
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WRKY transcription factors play important roles in many biological processes, and mainly bind to the W-box element to regulate gene expression. Previously, we characterized a WRKY gene from Tamarix hispida, ThWRKY4, in response to abiotic stress, and showed that it bound to the W-box motif. However, whether ThWRKY4 could bind to other motifs remains unknown. In this study, we employed a Transcription Factor-Centered Yeast one Hybrid (TF-Centered Y1H) screen to study the motifs recognized by ThWRKY4. In addition to the W-box core cis-element (termed W-box), we identified that ThWRKY4 could bind to two other motifs: the RAV1A element (CAACA) and a novel motif with sequence of GTCTA (W-box like sequence, WLS). The distributions of these motifs were screened in the promoter regions of genes regulated by some WRKYs. The results showed that the W-box, RAV1A, and WLS motifs were all present in high numbers, suggesting that they play key roles in gene expression mediated by WRKYs. Furthermore, five WRKY proteins from different WRKY subfamilies in Arabidopsis thaliana were selected and confirmed to bind to the RAV1A and WLS motifs, indicating that they are recognized commonly by WRKYs. These findings will help to further reveal the functions of WRKY proteins.
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Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Tamaricaceae/genética , Fatores de Transcrição/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Proteínas de Ligação a DNA/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Regiões Promotoras Genéticas/genética , Regiões Promotoras Genéticas/fisiologia , Tamaricaceae/metabolismo , Tamaricaceae/fisiologia , Fatores de Transcrição/fisiologia , Técnicas do Sistema de Duplo-HíbridoRESUMO
Ethylene responsive factors (ERFs) play important roles in the abiotic stress; however, only a few ERF genes from woody plants have been functionally characterized. In the present study, an ERF gene from Betula platyphylla (birch), BpERF11, was functionally characterized in response to abiotic stress. BpERF11 is a nuclear protein, which could specifically bind to GCC boxes and DRE motifs. BpERF11-overexpressing and BpERF11 RNA interference (RNAi) knockdown plants were generated for gain- and loss-of-function analysis. BpERF11 negatively regulates resistance to salt and severe osmotic stress, and the transgenic birch plants overexpressing BpERF11 shows increased electrolyte leakage and malondialdehyde (MDA) contents. BpERF11 inhibits the expression of an AtMYB61 homologous gene, resulting in increased stomatal aperture, which elevated the transpiration rate. Furthermore, BpERF11 downregulates the expression of P5CS, SOD and POD genes, but upregulates the expression of PRODH and P5CDH, which results in reduced proline levels and increased reactive oxygen species (ROS) accumulation. BpERF11 also significantly inhibits the expression of LEA and dehydrin genes that involve in abiotic stress tolerance. Therefore, BpERF11 serves as a transcription factor that negatively regulates salt and severe osmotic tolerance by modulating various physiological processes.
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Betula/genética , Etilenos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Osmorregulação/genética , Proteínas de Plantas/genética , Tolerância ao Sal/genética , Sequência de Aminoácidos , Betula/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Eletrólitos/metabolismo , Malondialdeído/metabolismo , Microscopia Confocal , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Filogenia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/classificação , Proteínas de Plantas/metabolismo , Estômatos de Plantas/genética , Estômatos de Plantas/fisiologia , Transpiração Vegetal/genética , Transpiração Vegetal/fisiologia , Plantas Geneticamente Modificadas , Prolina/metabolismo , Interferência de RNA , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de AminoácidosRESUMO
Plant zinc finger proteins (ZFPs) comprise a large protein family and they are mainly involved in abiotic stress tolerance. Although Arabidopsis RING/FYVE/PHD ZFP At5g62460 (AtRZFP) is found to bind to zinc, whether it is involved in abiotic stress tolerance is still unknown. In the present study, we characterized the roles of AtRZFP in response to abiotic stresses. The expression of AtRZFP was induced significantly by salt and osmotic stress. AtRZFP positively mediates tolerance to salt and osmotic stress. Additionally, compared with wild-type Arabidopsis plants, plants overexpressing AtRZFP showed reduced reactive oxygen species (ROSs) accumulation, enhanced superoxide dismutase and peroxidase activity, increased soluble sugars and proline contents, reduced K(+) loss, decreased Na(+) accumulation, stomatal aperture and the water loss rate. Conversely, AtRZFP knockout plants displayed the opposite physiological changes when exposed to salt or osmotic stress conditions. These data suggested that AtRZFP enhances salt and osmotic tolerance through a series of physiological processes, including enhanced ROSs scavenging, maintaining Na(+) and K(+) homeostasis, controlling the stomatal aperture to reduce the water loss rate, and accumulating soluble sugars and proline to adjust the osmotic potential.
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Recent work has shown that infrared stimulated luminescence (IRSL) signals in sedimentary coarse-grain K-feldspars are derived mainly from high temperature thermoluminescence (TL) peaks around 400°C, and the fading components of the IRSL signal can be preferentially removed by prior IR stimulation at relatively low temperature. Considering the complexity of TL signal for very old samples, we may choose non-fading components from K-feldspar TL signals using the combination of optical and thermal activation methods. This paper examines a protocol of post-IR isothermal TL (i.e. pIRITL) signal for sedimentary coarse-grain K-feldspars, which results from isothermal TL measurements following elevated temperature IR bleaching. We show that a sum of two exponential decay functions can fit well to the pIRITL decay curves, and both the holding temperature for isothermal TL measurements and the prior elevated temperature IR bleaching can affect greatly the fast components of pIRITL signal. The dose response ranges of pIRITL signal are wider than those of post-IR IRSL signals, but the relative high residual pIRITL signal means that it is not appropriate for dating young samples. It is expected that one isothermal TL signal for K-feldspar measured at ~400°C following IR bleaching at 290°C (i.e. pIRITL400) is useful for dating very old samples.