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In recent years, polysaccharides have received much attention because of their high safety and good immunological activity. The study of polysaccharide in vivo process is a key scientific problem that needs to be solved for polysaccharide drug development. Some progress has been made in the field of polysaccharide pharmacokinetics and immunomodulation. However, due to the lack of both chromogenic and light-absorbing groups and the complex molecular structure of polysaccharides, the in vivo processes and immunomodulatory mechanisms of polysaccharides have been slow to be investigated. The effective combination of multiple techniques can break the bottleneck of difficult tracing and unknown immunomodulatory mechanism of polysaccharides in vivo, and promote the development and utilization of polysaccharides. In this paper, we systematically summarize the key techniques in the study of polysaccharide in vivo processes and immunomodulatory mechanisms in order to provide technical references and research ideas for the study of polysaccharide in vivo processes and immunomodulatory mechanisms.
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Objective:To explore the mediating role of illness perception in the relationship between fear of disease progression and sleep quality in patients with ovarian cancer during chemotherapy, and to provide a theoretical basis for improving sleep quality in patients with ovarian cancer during chemotherapy.Methods:From January to August 2022, 300 patients with ovarian cancer undergoing chemotherapy in Shandong First Medical University Affiliated Provincial Hospital, Qilu Hospital of Shandong University, Shandong Cancer Hospital were included by convenient sampling. A cross-sectional questionnaire survey included the Chinese version of the Fear of Progression Questionnaire Short Form for cancer patients, the Brief Illness Perception Questionnaire, and the Pittsburgh Sleep Quality Index. Bivariate factor analysis, Spearman correlation analysis, and the Bootstrap confidence interval evaluation method were used.Results:A total of 287 valid questionnaires were collected. The scores of fear disease progression, illness perception, and sleep quality were 30.00 (22.00, 36.00), 37.00 (32.00, 44.00), and 6.00 (3.00, 11.00), respectively. Sleep quality was positively correlated with fear disease progression ( r=0.250, P<0.001) and illness perception ( r=0.326, P<0.001). Illness perception played a partial mediating role in the relationship between fear of disease progression and sleep quality, accounting for 41.4% of the total effect. Conclusions:Ovarian cancer patients during chemotherapy reported poor sleep quality. In clinical practice, health care providers including nurses can take interventions aimed at reducing fear of disease and improving illness perception level to improve the sleep quality of ovarian cancer patients during chemotherapy.
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The article summarized and analyzed the application status, specific application methods, shortcomings and prospects of mobile medical technology in the prevention and treatment of diabetic foot, and to provide reference for the application and future research of mobile medical technology in the prevention and treatment of diabetic foot in our country.
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Objective:To explore the relationship between challenge-hindrance stressors and thriving at work in clinical nurses, and to analyze the mediating role of intrinsic motivation.Methods:This was a cross-sectional survey. A total of 319 nurses from the Provincial Hospital Affiliated to Shandong First Medical University, Qilu Hospital of Shandong University, Shandong Provincial Qianfoshan Hospital from May to June 2022 were investigated by general data questionnaire, Challenge-Hindrance Stressors Scale, Intrinsic Motivation Scale and Thriving At Work Scale. Pearson was used to analyze the correlation between various variables, and Amos 23.0 was used to construct a structural equation model to analyze the mediating role of intrinsic motivation between challenging stressors, hindrance stressors, and thriving at work.Results:The score for challenging stressors was (21.22 ± 4.42) points, the score for hindrance stressors was (13.51 ± 3.59) points, the score for intrinsic motivation was (78.96 ± 11.52) points, and the score for thriving at work was (51.27 ± 8.03) points. Challenging stressors was positively associated with intrinsic motivation and thriving at work ( r=0.222, 0.221, both P<0.01), hindrance stressors was negatively associated with intrinsic motivation and thriving at work ( r=-0.152, -0.337, both P<0.01), intrinsic motivation was positively correlated with thriving at work ( r=0.564, P<0.01). Intrinsic motivation was partially mediated between challenging stressors, hindrance stressors and thriving at work, respectively accounting for 16.02% and 13.79%. Conclusions:Challenging stressors and hindrance stressors can indirectly influence their thriving at work through intrinsic motivation. Nursing managers should help nurses treat different stressors correctly to enhance their intrinsic motivation and promote their thriving at work.
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Objective:To explore the current status of discharge planning of ward nurses in cardiovascular department and analyzed influencing factors, and to provide reference for intervention strategies to build the discharge planning ability of the responsible nurses in cardiovascular department.Methods:This study was a cross-sectional survey. Convenience sampling method was adopted to select 267 ward nurses of cardiovascular department from 5 first-class hospitals in Dalian City from July to August 2022 as the research objects. The questionnaire survey was conducted by general data questionnaire, Job-Esteem Scale for Nurses in Hospital and Discharge Planning of Ward Nurses. Multiple linear regression analysis and Pearson correlation analysis were used for data analysis.Results:The total score of discharge planning of ward nurses in cardiovascular department was (103.86 ± 16.20) points; the total score of nurses' professional respect was (98.92 ± 13.67) points, and the total score of discharge planning of ward nurses in cardiovascular department was positively correlated with the total score of job-esteem and scores of all dimensions ( r values were 0.382-0.551, all P<0.01). Multiple linear regression analysis showed that age, professional title, certificate of cardiovascular nurses and job-esteem of nurses were influencing factors of discharge planning of ward nurses in cardiovascular department ( t values were 2.57-8.27, all P<0.05), accounting for 47.3% of the total variation. Conclusions:The discharge planning of ward nurses in cardiovascular department was above the medium level, and age, professional title, whether they were cardiovascular nurses and job-esteem were the influencing factors. From the perspective of improving nurses' job-esteem, nursing managers can focus on young nurses and nurses with low professional titles, attach importance to the training of specialized ward nurses in cardiovascular department, and take certain countermeasures to improve the discharge planning of ward nurses in cardiovascular department.
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Objective:To systematically evaluate the effect of repeated low-level red light (RLRL) therapy on controlling the progression of myopia in children.Methods:Medline, Embase, Cochrane Library, Web of Science, ClinicalTrial.gov and the CNKI, VIP, Wanfang database, and Chinese Clinical Trial Registry (www.chictr.org.cn) were searched from their establishment to September 2022 to obtain the randomized controlled trial (RCT) about RLRL controlling myopia progression in children.Grey literature was also searched.Inclusion and exclusion criteria were developed according to the PICOS principle.Two researchers screened the literature independently, extracted data, and assessed the risk of bias in the included studies using the Cochrane collaboration tool for assessing risk of bias.The data were imported into RevMan 5.4 software for meta-analysis.Mean and standard deviation of data were used to obtain mean differences and 95% confidence intervals of included studies.Changes in spherical equivalent (SE) and axial length (AL) were compared between before and after the intervention in RLRL and single-vision spectacle (experimental group) and single-vision spectacle (control group). The GRADE system was used to grade the quality of evidence for the outcome indicators.Results:A total of 157 studies were obtained.After the screening, 7 RCT studies including 1 038 study subjects met the inclusion criteria.Meta-analysis results showed that retarded SE progression was retarded and AL elongation was reduced significantly after different follow-up durations (1, 3, 6, 12 and 24 months) in experimental group compared with control group.As the follow-up duration extended from 1 month to 24 months, the retard of SE progression increased from 0.14 to 0.93 D, and the reduction of AL increased from 0.07 to 0.48 mm.Both 635 and 650 nm red light significantly retarded SE progression and reduced AL elongation.The two outcome indicators, SE change and AL change were moderate-strength evidence by GRADE.Conclusions:The current moderate-strength evidence shows that RLRL can better retard myopia progression in children than single-vision spectacle glasses.
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Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.
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To understand the current situation of occupational burnout, turnover intention, and professional self-care among social workers under the context of epidemic prevention and control, and the role of professional self-care in the relationship between occupational burnout and turnover intention, this study investigated 1 066 social workers in Fujian Province with Professional Self-Care Practice Scale, and the Burnout Scale. SPSS 26.0 and its PROCESS plug-in analysis were used, the results showed that the proportion of social workers with moderate and severe occupational burnout accounted for 44.22% and 29.99% respectively, the proportion with turnover intention in the past year accounted for 25.67%, and the average score of professional self-care was (22.57±5.98) points. There was a significant positive correlation between occupational burnout and turnover intention (r=0.29, P<0.01). Professional self-care of social workers was significant negatively correlated with occupational burnout (r=-0.46, P<0.01) and turnover intention (r=-0.16, P<0.01). Professional self-care played a moderating role in the relationship between occupational burnout and turnover intention (β=-0.01, P<0.05). This paper indicated that social workers had a high degree of occupational burnout and turnover intention, low level of professional self-care under the context of epidemic prevention and control, and the attention should be paid to the protective role of professional self-care.
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Aim To study the hypnotic effect and safety of compound anshen essential oil. Methods Gas chromatograph-mass spectrometer (GC-MS) was used to analyze the main active components of compound anshen essential oil. The mouse model of insomnia was established by intraperitoneal injection of para-chloro-phenyl alanine ( PC PA ) , combined with pentobarbital sodium sleep experiment and EEG characteristic monitoring in rats to study the hypnotic effect and mechanism. The safety of compound anshen essential oil was evaluated by acute toxicity test, skin irritation/allergy test and 90-day repeated administration toxicity test. The clinical effect and safety were evaluated by using the sleep monitoring technology for micro-motion sensitive mattress. Results Four components, including Atractylone (34.61%), (+) -Limonene (17.80%) , Linalool (11.63%), and Ocimene (11.67%) , were detected as the main active components of compound anshen essential oil. Compound anshen essential oil in-halation administration for seven days could effectively reduce the autonomic activity of insomnia mice, shorten the sleep latency (P <0.05) , improve the sleep duration, increase of neurotransmitters such as 5-hydroxy tryptamine (5-HT) and -γ-aminobutyric acid (GABA) in brain of mice with insomnia, and the medium dose group had better hypnotic effect. There was no death or adverse reaction in the safety evaluation test. The sleep balance index of 10 subjects with difficulty in falling a-sleep significantly increased (P <0.05), sleep latency was significantly shortened (P <0.05) , total sleep duration and sleep efficiency were improved, and no ad¬verse reactions were found after using the compound anshen essential oil for two days. Conclusions The compound anshen essential oil developed by the research team is safe and effective in relieving sleep disorders, which may be closely related to the co-regulation of the levels of neurotransmitters such as 5-HT and GABA by the four main active components.
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Myocardial fibrosis is a common pathological feature in various advanced cardiovascular diseases, and progressive fibrosis is the pathological basis for the development and progression of many cardiac arrhythmias and heart failure. There are no effective reversal drugs for myocardial fibrosis, which is related to the lack of understanding of the molecular mechanisms. Noncoding RNAs are a class of RNAs that do not function as coding proteins, and have been found to be intimately involved in the life cycle of cardiomyocyte differentiation, transcription and apoptosis, and are important regulators of cardiovascular diseases. An increasing number of studies have shown that noncoding RNAs regulate the proliferation and transformation of cardiac fibroblasts through related signaling pathways and can be used as potential biomarkers and novel therapeutic targets for cardiac fibrosis. This article reviews the relationship between noncoding RNAs and cardiac fibrosis.
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This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.
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Antioxidantes/química , Simulação de Acoplamento Molecular , Artemisia , Farmacologia em Rede , Fosfatidilinositol 3-Quinases , Anti-Inflamatórios/química , Medicamentos de Ervas Chinesas/farmacologia , Interleucina-6RESUMO
OBJECTIVE@#To investigate the mechanism by which fibroblasts with high WNT2b expression causes intestinal mucosa barrier disruption and promote the progression of inflammatory bowel disease (IBD).@*METHODS@#Caco-2 cells were treated with 20% fibroblast conditioned medium or co-cultured with fibroblasts highly expressing WNT2b, with the cells without treatment with the conditioned medium and cells co-cultured with wild-type fibroblasts as the control groups. The changes in barrier permeability of Caco-2 cells were assessed by measuring transmembrane resistance and Lucifer Yellow permeability. In Caco-2 cells co-cultured with WNT2b-overexpressing or control intestinal fibroblasts, nuclear entry of β-catenin was detected with immunofluorescence assay, and the expressions of tight junction proteins ZO-1 and E-cadherin were detected with Western blotting. In a C57 mouse model of dextran sulfate sodium (DSS)-induced IBD-like enteritis, the therapeutic effect of intraperitoneal injection of salinomycin (5 mg/kg, an inhibitor of WNT/β-catenin signaling pathway) was evaluated by observing the changes in intestinal inflammation and detecting the expressions of tight junction proteins.@*RESULTS@#In the coculture system, WNT2b overexpression in the fibroblasts significantly promoted nuclear entry of β-catenin (P < 0.01) and decreased the expressions of tight junction proteins in Caco-2 cells; knockdown of FZD4 expression in Caco-2 cells obviously reversed this effect. In DSS-treated mice, salinomycin treatment significantly reduced intestinal inflammation and increased the expressions of tight junction proteins in the intestinal mucosa.@*CONCLUSION@#Intestinal fibroblasts overexpressing WNT2b causes impairment of intestinal mucosal barrier function and can be a potential target for treatment of IBD.
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Humanos , Camundongos , Animais , Células CACO-2 , beta Catenina/metabolismo , Meios de Cultivo Condicionados/farmacologia , Junções Íntimas/metabolismo , Mucosa Intestinal , Doenças Inflamatórias Intestinais , Proteínas de Junções Íntimas/metabolismo , Inflamação/metabolismo , Fibroblastos/metabolismo , Camundongos Endogâmicos C57BL , Glicoproteínas/metabolismo , Proteínas Wnt/farmacologia , Receptores Frizzled/metabolismoRESUMO
Objective: To explore the mechanism of intestinal tissue damage induced by macrophages activated by WNT2B high-expressed fibroblasts. Methods: This study involved biological information analysis, pathological tissue research and cell experimental research. The biological information of the colon tissue from the children with inflammatory bowel disease in previous study was analyzed again with single-cell sequencing. The pathological tissues were collected by colonoscopy from 10 children with Crohn's disease treated in the Department of Gastroenterology of Guangzhou Women and Children's Medical Center from July 2022 to September 2022. According to the findings of colonoscopy, tissues with obvious inflammation or ulceration were classified as the inflammatory group, while tissues with slight inflammation and no ulceration were classified as the non-inflammatory group. HE staining was performed to observe the pathological changes of the colon tissues. Macrophage infiltration and CXCL12 expression were detected by immunofluorescence. In terms of cell experiments, fibroblasts transfected with WNT2B plasmid or empty plasmid were co-cultured with salinomycin treated or non-treated macrophages, respectively; the expression of proteins through Wnt classical pathway were detected by western blotting. Macrophages treated with SKL2001 were used as the experimental group, and those with phosphate buffer as the control group. The expression and secretion of CXCL12 in macrophages were detected by quantitative Real-time PCR and enzyme-linked immunosorbent assay (ELISA). T-test or rank sum test were used for the comparison between groups. Results: Single-cell sequencing analysis suggested that macrophages were the main cells in inflammatory bowel disease colon tissue, and there was interaction between WNT2B high-expressed fibroblasts and macrophages. HE staining of the 10 patients ((9.3±3.8) years old, 7 males and 3 females) showed that the pathological score of colon tissue in the inflammatory group was higher than that in the non-inflammatory group (4 (3, 4) vs. 2 (1, 2) points, Z=3.05, P=0.002). Tissue immunofluorescence indicated that the number of infiltrating macrophages in the inflammatory group was significantly higher than that in the non-inflammatory group under high power field of view (72.8±10.4 vs.8.4±3.5, t=25.10, P<0.001), as well as the number of cells expressing CXCL12 (14.0±3.5 vs. 4.7±1.9, t=14.68, P<0.001). In cell experiments, western blotting suggested an elevated level of glycogen synthase kinase-3β phosphorylation in macrophages co-cultured with fibroblast transfected with WNT2B plasmid, and salinmycin could reverse this change. Real-time PCR suggested that the transcription level of CXCL12 in the experimental group was higher than that in the control group (6.42±0.04 vs. 1.00±0.03, t=183.00, P<0.001), as well as the expression and secretion of CXCL12 by ELISA ((465±34) vs. (77±9) ng/L, t=13.21, P=0.006). Conclusion: WNT2B high-expressed fibroblasts can secrete WNT2B protein and activate the Wnt classical signaling pathway thus enhancing the expression and secretion of CXCL12 in macrophages, inducing the development of intestinal inflammation of Crohn's disease.
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Criança , Masculino , Humanos , Feminino , Pré-Escolar , Adolescente , Doença de Crohn , Doenças Inflamatórias Intestinais , Colo , Inflamação , Colonoscopia , Glicoproteínas , Proteínas WntRESUMO
Aim To explore the inhibitory effect of lenvatinib plus fluvastatin on liver transplantation tumor in mice and the mechanism.Methods Mouse model of subcutaneous liver cancer was used.Single agent of lenvatinib, single agent of fluvastatin, a combination of lenvatinib and fluvastatin and control solvent were given to four groups of mice.Tumor volume was measured.Immunohistochemistry was used to examine proliferation of tumor cells.Tunel was employed to detect the cell apoptosis.qRT-PCR and immunohistochemistry were used to measure the expression of TLR4.Western blot was employed to determine β-catenin expression.Rescue experiment was done using human hepatoma cells cultured in vitro.Results Treatment with both lenvatinib and fluvastatin significantly suppressed tumor growth in nude mice.Combined treatment significantly decreased the expressions of PNCA and increased apoptosis in tumor cells.Mechanically combined treatment synergistically suppressed the mRNA and protein expression of TLR4 which further inhibited the expression of β-catenin in hepatoma cells.Conclusions A combination of lenvatinib and fluvastatin synergistically inhibits tumor growth and promotes tumor cell apoptosis.The combination treatment significantly inhibits TLR4/β-catenin signaling pathway.
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This article reviews the concept, evaluation criteria, current situation, influencing factors and the impact on health of intrinsic capacity in the elderly at home and abroad, in order to provide reference for clinical medical staff to improve the elderly′s intrinsic capacity, and actively carry out targeted interventions and related research.
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Objective:To explore the value of radiomics model based on intratumoral and peritumoral early dynamic contrast-enhanced (DCE) MRI for identifying benign and malignant in breast imaging reporting and data system (BI-RADS) 4 tumors.Methods:A total of 191 patients diagnosed with BI-RADS 4 breast tumors by breast MRI examination with clear pathological diagnosis from January 2016 to December 2020 in the First Affiliated Hospital of Bengbu Medical College were analyzed retrospectively, including 77 benign and 114 malignant cases, aged 23-68 (46±10) years. The one-slice image with the largest area of the lesion of the second stage DCE-MRI images was selected to outline the region of interest, and automatically conformal extrapolated by 5 mm to extract the intra-tumoral and peritumoral radiomics features. The included cases were randomly divided into training and testing cohorts in the ratio of 8∶2. The statistical and machine learning methods were used for feature dimensionality reduction and selection of optimal radiomics features, and logistic regression was used as the classifier to establish the intratumoral, peritumoral, and intratumoral combined with peritumoral radiomics models. The independent risk factors that could predict the benignity and malignancy of breast tumors were retained as clinical-radiological characteristics by univariate and multivariate logistic regression to establish a clinical-radiological model. Finally, the intratumoral and peritumoral radiomics features were combined with clinical-radiological features to develop a combined model of the three. The receiver operating curve was used to analyze the predictive performance of each model and calculate the area under the curve (AUC),the AUC was compared by DeLong test. The stability of the three-component combined diagnostic model was tested by 10-fold cross-validation, and the model was visualized by plotting nomogram and calibration curves.Results:In the training cohort, the AUC of the three-component combined model for identifying benign and malignant BI-RADS 4 breast tumors was significantly higher than that of the intratumoral radiomics model ( Z=3.38, P<0.001), the peritumoral radiomics model ( Z=4.01, P<0.001), the intratumoral combined with peritumoral radiomics model ( Z=3.11, P=0.002), and the clinical-radiological model ( Z=3.24, P=0.001). And the AUC, sensitivity, specificity, accuracy, and F1-score of the three-component combined model were 0.932, 91.2%, 86.9%, 87.0% and 0.89, respectively. In the testing cohort, the three-component combined model also had the highest AUC value (0.875), and diagnostic sensitivity, specificity, accuracy and malignancy F1-score were 95.7%, 62.5%, 76.9%, and 0.89, respectively. The AUC calculated by 10-fold cross-validation was 0.90 (0.85-0.92), and the predicted curve of the three-component combined model in the calibration curve was in good agreement with the ideal curve. Conclusion:The three-component combined diagnostic model based on the intratumoral and peritumoral radiomics features and clinical-radiological features of early DCE-MRI has good performance and stability for identifying the benign and malignant in BI-RADS 4 breast tumors, and it can provide guidance for clinical decision non-invasively.
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Objective:To investigate the clinical significance of serum 25-hydroxyvitamin D [25(OH)D] level in patients with connective tissue disease (CTD) related-pulmonary arterial hypertension (PAH).Methods:CTD patients with PAH (CTD-PAH) and without PAH (CTD-non-PAH) were colle-cted. All data were analyzed.Results:The serum 25(OH)D in the CTD-PAH group was significantly lower than that in the CTD-non-PAH group [(14±8) ng/ml vs (20±8) ng/ml, t=-5.94, P<0.001]. The 25(OH)D deficiency rate in the CTD-PAH group 86.2%(112/130) was significantly higher than that in the CTD-non-PAH group 57.7% (75/130) ( χ2=26.07, P<0.001), while the insufficiency rate was significantly lower [10.0%(13/130) vs 32.3% (42/130), χ2=19.39, P<0.001]. Serum 25(OH)D levels in the systemic lupus erythematosus (SLE), systemic sclerosis (SSc) associated PAH group were lower than those in the SLE [14(8, 17) ng/ml vs 19(15, 23) ng/ml, Z=-3.66, P<0.001], SSc [11(8, 17) ng/ml vs 24(18, 30) ng/ml, Z=-4.97, P<0.001] without PAH group. The levels of serum 25(OH)D in CTD-PAH youthful group, in the middle age group were lower than that in CTD-non-PAH youthful group [(12±8) ng/ml vs (19±8) ng/ml, t=-4.36, P<0.001] and in the middle age group [(14±7) ng/ml vs (21±8) ng/ml, t=-3.75, P<0.001]. Serum levels of 25(OH)D [ OR (95% CI)=1.100 (1.058, 1.144), P<0.001], uric acid [ OR(95% CI)=0.996(0.993, 0.998), P=0.003], immune globulin (Ig)G [ OR(95% CI)=1.123(1.057, 1.194), P<0.001] were associated with PAH in CTD patients. Serum 25(OH)D was positively correlated with calcium ( r=0.24, P=0.007), while negatively correlated between serum 25(OH)D and IgM ( r=-0.34, P<0.001). Conclusion:The occurrence and development of CTD-PAH may be related to the decrease of 25(OH)D level. Serum 25(OH)D level is associated with PAH in CTD patients.
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Objective: To express DNA-binding protein (DBP) of human adenovirus (HAdV) type 7 using the prokaryotic expression system, and product anti-HAdV-7 DBP rabbit polyclonal antibody. Methods: The HAdV-7 DBP gene was synthesized and cloned into prokaryotic expressing vector pET30a, and the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cell. The recombinant protein DBP was expressed by induced Isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with Ni-NTA affinity column. The titer of anti-DBP polyclonal antibody produced in immunized rabbit was measured by indirect ELISA, and the specificity of the antibody was identified by Western blotting and indirect immunofluorescence assay (IFA). In addition, purified rDBP was used as coating antigen for indirect ELISA assay to detect specific IgM and IgG antibodies against DBP in the serum of children infected with HAdV. Results: The HAdV-7 DBP plasmid was constructed successfully. The purified recombinant DBP was more than 95% after purification. The titer of polyclonal antibody was 1∶1 024 000. The polyclonal antibody showed high specificity in vitro using Western blotting and IFA. The positive rate of specific anti-DBP IgM and IgG antibody in acute-phase serum samples collected from children infected with HAdV were 50.0% (19/38) and 63.2% (24/38), respectively, using indirect ELISA. Conclusion: In summary, the HAdV-7 rDBP is expressed using prokaryotic expression system, and the recombinant HAdV-7 DBP protein and the anti-DBP rabbit polyclonal antibody with high titer are prepared.
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Animais , Coelhos , Adenovírus Humanos/genética , Especificidade de Anticorpos , Western Blotting , Proteínas de Ligação a DNA/metabolismo , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Imunoglobulina GRESUMO
ObjectiveTo establish the frozen section method of Ferula ferulaeoides, and to study the histochemical localization of volatile oil and coumarins in different organs of F. ferulaeoides. MethodThe roots, stems, petioles and leaves of F. ferulaeoides were used as materials to investigate the concentration of sucrose protectant, liquid nitrogen flash-freezing time, embedding conditions, section thickness, freezing temperature and time and post-treatment methods, the most suitable section conditions were screened by comparing the integrity, microscopic effect, elongation and clarity of frozen sections. Sudan Ⅳ staining method and fluorescence microscopy were used to locate the volatile oil and coumarins of F. ferulaeoides. ResultThe optimal conditions for frozen sections of the roots, stems, petioles and leaves of F. ferulaeoides were as follows:10%, 15% and 20% gradient sucrose as the protectant for roots, 10%, 20% and 30% gradient sucrose as the protectant for stems and petioles, 20%, 25% and 30% gradient sucrose as the protectant for leaves, glue-water (2∶1) as the embedding agent, quick-freeze in liquid nitrogen for 20 s, warmed up at -25 ℃ for 30 min, sliced at -20 ℃ with the thickness of 25 μm, rinsed with the same concentration of sucrose solution (gradient sucrose solution selected the last concentration), and the slices placed on the ice pack for a period of time and stored at room temperature. Among them, the concentration of sucrose protectant was the most important factor. The results of histochemical localization showed that volatile oil and coumarins in four organs of F. ferulaeoides were mainly distributed in resin canal. ConclusionFrozen section of F. ferulaeoides is established for the first time with high rate of slicing and simplified steps, its volatile oil and coumarins are mainly accumulated in resin canal.
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Background@#Assessing nuclear features is diagnostically challenging in the aspect of thyroid pathology. The aim of this study was to determine whether pathologists could distinguish BRAF-like and RAS-like nuclear features morphologically and identify morphological features to differentiate thyroid tumors with RAS-like mutations from encapsulated papillary thyroid carcinoma (PTC) with predominant follicular growth and BRAFV600E mutation. @*Methods@#Representative whole slide images of 16 encapsulated thyroid tumors with predominant follicular growth were reviewed by 12 thyroid pathologists using a web browser-based image viewer. Total nuclear score was calculated from semi-quantitatively scored eight nuclear features. The molecular profile of RAS and BRAF genes was determined by Sanger sequencing. @*Results@#Total nuclear score ranging 0 to 24 could differentiate BRAF-like tumors from RAS-like tumors with a cut-off value of score 14. The interobserver agreement was the highest for the assessment of nuclear pseudoinclusions (NPIs) but the lowest for nuclear elongation and sickle-shaped nuclei. NPIs were found in tumors with BRAFV600E mutation, but not in tumors with RAS-like mutations. Total nuclear scores were significantly higher for tumors with BRAFV600E than for those with RAS-like mutations (P<0.001). @*Conclusion@#Our results suggest that NPIs and high nuclear scores have diagnostic utility as rule-in markers for differentiating PTC with BRAFV600E mutation from benign or borderline follicular tumors with RAS-like mutations. Relaxation of rigid criteria for nuclear features resulted in an overdiagnosis of PTC. Immunostaining or molecular testing for BRAFV600E mutation is a useful adjunct for cases with high nuclear scores to identify true PTC.