Objective: To investigate the environmental contamination of norovirus in nurseries and primary/secondary schools, so as to provide a scientific basis for effective prevention and control measures. Methods: A total of 483 external environmental samples were collected from 34 cluster outbreaks of norovirus gastroenteritis in kindergartens and primary/secondary schools in Linhai City from 2021 to 2024. Pathogen detection was conducted using a rapid nucleic acid extraction kit and realtime fluorescence RT-PCR, and the results were analyzed using the χ2 test or Fishers exact test. Results: Among the collected external environmental samples, the total positive rate of surface contamination was 13.66%. The positive rates in kindergartens and primary/secondary schools were 12.20% and 15.82%, respectively. In kindergartens, the five surfaces with the highest detection rates were desks/chairs (23.33%), toilet stool troughs (20.69%), urinal troughs (12.00%), washbasins/sinks (11.11%), and toilet mops (9.38%). In primary/secondary schools, the top five were toilet stool troughs (38.30%), urinal troughs (23.53%), toilet door handles (13.04%), toilet mops (12.50%), and drinking cups (11.11%). The difference in positive detection rates among different external environments in primary/secondary schools was statistically significant (Fishers exact probability test, P<0.01). The positive detection rate in sanitary toilets was higher than that in classroom environments (χ2=17.38), while the positive detection rate in classroom environments of kindergartens was higher than that in primary/secondary schools (χ2=5.42)(P<0.05). Conclusions Norovirus exhibits a high contamination rate in nurseries and schools, particularly in restroom areas. Strengthening sanitation and disinfection in highrisk environments, and improving hygiene awareness among children and staff, are essential for the effective prevent and control of norovirus.

Objective To develop an evidence-based practice program for ICU eye care and evaluate its application effect using Stetler model of research utilization.Methods The systematic search was carried out in domestic and foreign guid elines,systematic reviews,evidence summaries,expert consensuses on eye care for ICU patients.The quality of the included literature was evaluated and the evidence was summarized.The evidence based on expert advice,clinical context and stakeholder opinions was screened.According to Stetler model of research utilization,the evidence translation/application was formulated from 3 aspects,including method,level and type of evidence application,and the evidence translation was implemented using a stepwise trial design.Five ICUs in a tertiary hospital in Zhejiang Province were selected,and the order of enrollment of each ICU was determined by computer-generated random numbers from April to september,2022.In the first month,all ICUs were in pre-evidence-based practice period,and in the second month,an ICU was trained and entered the evidence-based practice period.Until the 6th month,all ICUs entered the evidence-based practice period.Then the rate of ocular symptoms and implementation rate of eye care measures were compared before and after EBP.Results A total of 1 540 patients were included.With regard to the implementation rate of eye care measures,the rate of lubricant use and suspected ocular infection management in patients at high risk for ocular complications in evidence-based practice period was significantly higher compared with pre-evidence-based practice period(P＜0.001),and no statistically significant difference was found in the rate of eyelid closure.The incidence of conjunctival exposure,subconjunctival hemorrhage,conjunctival congestion,and ocular discharge in non-neurology critical care patients decreased compared with pre-evidence-based practice period(P＜0.05),and the incidence of conjunctival edema and conjunctival congestion in NICU patients decreased compared with pre-evidence-based practice period as the intervention continued(P＜0.001).Conclusion Evidence-based practice in eye care for ICU patients using Stetler model of research utilization can standardize the practice of eye care for ICU patients and improve the quality of care.

BACKGROUND: The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period. METHODS: Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7. RESULTS: Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs. CONCLUSION The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

Objective:To investigate the role and diagnostic value of miRNA-205 in chronic kidney disease (CKD) patients with vascular calcification.Methods:It was divided into in vitro cell experiment and retrospective cohort study. In vitro experiments were conducted by using rat thoracic aortic smooth muscle cells. Alizarin red staining and calcium content detection were used to detect the calcification of vascular smooth muscle cells (VSMCs). Alkaline phosphatase (ALP) test kit was used to measure ALP activity. Western blotting was used to detect the protein expression levels of osteogenic transcription factors runt-related transcription factor 2 (Runx2), α smooth muscle actin (α-SMA) and smooth muscle-22α (SM-22α) in VSMCs. qRT-PCR was used to detect miRNA-205 and Runx2 expression levels. The double luciferase reporter gene assay was used to verify the targeted relationship between miRNA-205 and Runx2. The non-dialysis patients with CKD 3-5 stage from June 2020 to January 2021 in the Department of Nephrology of Fourth Hospital, Hebei Medical University were selected. According to coronary artery calcium score (CACs), the patients were divided into non-calcification group (CACs=0), mild-moderate calcification group (0 400). Spearman correlation analysis was used to analyze the correlation between miRNA-205 and Runx2 and vascular calcification. Logistic regression model and receiver operating characteristic (ROC) curve analysis were used to analyze the ability of miRNA-205 to predict the vascular calcification in patients with CKD. Results:(1)Compared with the control group, calcium nodules were more, and the calcium content, ALP activity and Runx2 protein level were higher, and the expression levels of miRNA-205, α-SMA and SM-22α were significantly lower in high phosphorus group (all P<0.05). Overexpression of miRNA-205 significantly reduced the calcification of VSMCs and Runx2 protein level, and increased the protein levels of α-SMA and SM-22α (all P<0.05). miRNA-205-5p reduced the activity of luciferase in the wild-type Runx2-3'-end non-coding region plasmid. (2) Eighty CKD patients were enrolled, with age of (57.50±14.93) years old and 49 males (61.3%). The results of comparison of miRNA-205 and Runx2 expression levels in non-calcification group ( n=26), mild- moderate calcification group ( n=30) and severe calcification group ( n=24) showed that, the higher degree of calcification, the lower miRNA-205 expression level and the higher Runx2 mRNA expression level (all P<0.05). miRNA-205 was negatively correlated with CACs ( r=-0.50, P<0.01) and Runx2 was positively correlated with CACs ( r=0.55, P<0.01). Multivariate logistic regression analysis results suggested that miRNA-205 ( OR=0.451, 95% CI 0.122-0.873) was an independent influencing factor of vascular calcification in CKD patients. The area under the ROC curve of miRNA-205 and miRNA-205 combined with Runx2 for predicting vascular calcification were 0.796 (95% CI 0.697-0.859) and 0.924 (95% CI 0.866-0.982), respectively. Conclusions:miRNA-205 inhibits vascular calcification by targeting Runx2 to negatively regulate osteogenetic phenotype transformation of VSMCs and is expected to be an early diagnostic marker of vascular calcification in CKD patients.

Objective:To know the current situation of kinesiophobia in patients after cardiac surgery under cardiopulmonary bypass, and to clarify its influencing factors, so as to provide reference for developing intervention strategies to improve kinesiophobia level.Methods:This was a cross-sectional study. From February 2022 to September 2022, the patients after cardiac valve surgery under cardiopulmonary bypass in the Second Affiliated Hospital Zhejiang University School of Medicine were investigated by convenience sampling methods. The survey was conducted using the General Information Questionnaire, The Tampa Scale for Kinesiophobia Heart, Exercise Self-Efficacy Scale, Pain Catastrophizing Scale, and Adaptation, Partnership, Growth, Affection and Resolve(APGAR) as research tools, and the influencing factors were analyzed using univariate and binary Logistic regression analysis.Results:A total of 219 patients were included, of which 97 patients (44.3%) had kinesiophobia. The results of binary Logistic regression analysis showed that monthly family income level, first time out of bed after operation, fear of falling, the family APGAR, and pain catastrophizing were significant influencing factors of kinesiophobia in patients after heart valve surgery under cardiopulmonary bypass (all P<0.05). Conclusions:The prevalence of kinesiophobia is high among patients after heart valve surgery under cardiopulmonary bypass. Clinicians should pay attention to patients with low monthly family income level, late first time out of bed after surgery, and fear of falling, as well as strengthen communication with patients and families, focus on the management of acute postoperative pain. In order to reduce or avoid the occurrence of kinesiophobia and enable patients to benefit from early ambulation.

OBJECTIVE To explain the material basis and biological mechanism of Astragali Radix’s “invigorating Qi” effect to regulate energy metabolism. METHODS The TCMSP database and literature search collected potential active components of Astragali Radix, the SEA database performed target prediction based on structural similarity, and the GeneCards, OMIM, and TTD databases obtained energy metabolism targets. Cytoscape software was used to construct protein-protein interaction network maps of Astragali Radix regulated energy metabolism targets, and GO and KEGG enrichment analyses were performed. Molecular docking and hierarchical cluster analysis were performed to evaluate the target-component affinity between the whole constituents of Astragali Radix and key targets, and the effects of representative compounds of Astragali Radix on the energy metabolism of H9C2 cardiomyocytes and GES-1 gastric epithelial cells were detected, and the binding mode analysis was conducted. RESULTS Network pharmacology results showed that there were 126 potential targets of Astragali Radix regulating energy metabolism. GO and KEGG enrichment analysis showed that Astragali Radix regulating energy metabolism might be related to gene expression of oxidation-reduction process, protein and enzyme synthesis. Among them, SIRT1 and PPARγ were key targets involved in the regulation of energy metabolism. Molecular docking and hierarchical clustering showed that Astragali Radix components had superior targeting to SIRT1 and PPARγ, and three representative compounds were selected for in vitro experimental verification in combination with molecular docking scores. Quercetin and kaempferol could promote energy metabolism in H9C2 cardiomyocytes and GES-1 gastric epithelial cells. The binding mode analysis showed that quercetin and kaempferol had preferable binding ability to SIRT1 and PPARγ. CONCLUSION In this study, the material basis and biological mechanism of Astragali Radix regulating energy metabolism are preliminarily explained by traditional Chinese medicine chemo-bio informatics methods, which provide a scientific basis for the connotation of Astragali Radix exerting the effect of stagnation and arthralgia through “invigorating Qi” in traditional Chinese medicine.

OBJECTIVE To investigate the cellular uptake and in vivo imaging of paclitaxel(PTX)-loaded polyethylene glycol(PEG)-modified rhein conjugate micelles. METHODS (P4/P2+PTX)/CRmP micelles were prepared by co-loading environment-responsive fluorescent probes P4/P2 and PTX into CRmP micelles. Using MCF-7 cells as cell models, the uptake of the micelles by MCF-7 cells was analyzed by laser confocal microscopy and flow cytometry. The H22 subcutaneous transplanted tumor mice were used as animal models, and the distribution of the micelles in vivo and in vitro were analyzed by in vivo imaging system. RESULTS (P4+PTX)/CRmP micelles in the intact form were internalized by MCF-7 cells and distributed in the cytoplasm. (P2+PTX)/CRmP micelles were more accumulated in the liver and tumor sites in vivo, and gradually accumulated at the tumor sites during the experimental time. CONCLUSION The intact micelles can be taken up by tumor cells, and have liver and tumor targeting properties in vivo.

Objective:To investigate the normal reference values of spinal bone mineral density measured by quantitative computed tomography (QCT) and the differences of bone mineral density (BMD) in different regions of in Chinese adult males.Methods:Men who underwent low-dose CT lung scan for cancer screening in regions of Northeast, North, East, South, Central and Southwest of China from January 2018 to December 2019 were selected. And the lumbar vertebrae BMD values in the male subjects were measured by the QCT system (Mindways Software, Inc.). The mean BMD values and their decline rates were calculated at an age interval of 10 years, and the prevalence of osteoporosis was calculated according to the American College of Radiology spine QCT osteoporosis diagnostic criteria.Results:A total of 50 682 males with a mean age of (50.22±12.79) years (ranged 20 to 98 years) were included in this study. The peak BMD of (173.11±28.56) mg/cm 3 in the healthy Chinese adult male population appeared in the age group of 20 to 29 years and then declined with age. Before the age of 70 years, the BMD was relatively higher in males in South China, and it was lower in Central China and Southwest China, and it was intermediate in Northeast, North and East of China, with statistically significant differences. There was no significant differences in BMD in the males in the two age groups of 70 to 79 years and 80 and older among the regions in China. The overall decline rate of spinal BMD in Chinese males under QCT was about 46.92% over the lifetime, and it declined obviouslyin the 40-49 age group. The overall prevalence of osteoporosis in Chinese male population aged 50 years and above was approximately 11.42%, with the highest prevalence in Southwest China and Central China (14.72% and 13.87%, respectively) and the lowest in North China and South China (8.53% and 7.71%, respectively). Conclusions:A reference of lumbar spine BMD values for healthy males in China based on QCT is established. BMD values were highest in South China and Lowest in Central China.

【Objective】 To establish and optimize a method for the determination of aluminum (Al) residue by inductively coupled plasma mass spectrometry (ICP-MS). 【Methods】 Nitric acid solution was used to treat samples and standards. The concentration of nitric acid solution and equipment parameters were optimized, and the specificity, linearity, repeatability, accuracy, detection limit, quantitative limit and intermediate precision of the optimized detection method were investigated to confirm whether it was suitable for the determination of Al residue in human serum albumin. 【Results】 The concentration of nitric acid was 5%, and digest time was 4 h. The equipment condition of ICP-MS was as follows: RF power: 1600 W, sampling depth: 10 mm, atomizer / carrier gas flow rate: 1.0 L/ min, compensation flow rate: 0.5 L/ min, experimental mode: standard mode, integration time: 0.2 s, data acquisition: 3 times. Specificity: The recoveries of Al: 92% (high concentration, RSD=3.5%), 98% (low concentration, RSD=4.9%). Linearity: In the range of (0~40) μg/L, the correlation coefficient between concentration and optical energy signal (CPS) of standard / sample were higher than 0.999 0. Accuracy/ Repeatability: The recoveries of sample (3 concentration): 108% (RSD=4.7%), 110% (RSD=4.9%) and 110% (RSD=2.8%). The detection limit was 0.006 μg/L, and the quantitation limit was 0.019 μg/L. Intermediate precision: personnel factor and date factor, P>0.05, RSD (12 times)=2%. Comparison between ICP-MS and atomic absorption spectrometry (AAS): the deviation between ICP-MS and AAS was 8%, and that of samples was 3%, with no significant difference noticed between the two methods. 【Conclusion】 After optimization, ICP-MS method has shown good performance in terms of specificity, linearity, repeatability, accuracy, detection limit, quantitative limit and intermediate precision, and is suitable for the determination of Al residue in human albumin products of our company.