OBJECTIVE: To identify the role of gene silencing or overexpression of Nemo-like kinase (NLK) during the process of neural differentiation of human mesenchymal stem cells (hBMSCs), and to explore the effect of NLK downregulation by transfection of small interfering RNA (siRNA) on promoting neuralized tissue engineered bone regeneration. METHODS: NLK-knockdown hBMSCs were established by transfection of siRNA (the experimental group was transfected with siRNA silencing the NLK gene, the control group was transfected with control siRNA and labeled as negative control group), and NLK-overexpression hBMSCs were established using lentivirus vector transfection technique (the experimental group was infected with lentivirus overexpressing the NLK gene, the control group was infected with an empty vector lentivirus and labeled as the empty vector group). After neurogenic induction, quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression of neural-related gene, and Western blot as well as immunofluorescence staining about several specific neural markers were used to evaluate the neural differentiation ability of hBMSCs.6-week-old male nude mice were divided into 4 groups: ① β-tricalcium phosphate (β-TCP) group, ② β-TCP+ osteogenic induced hBMSCs group, ③ β-TCP+ siRNA-negative control (siRNA-NC) transfection hBMSCs group, ④ β-TCP+ siRNA-NLK transfection hBMSCs group. Four weeks after the subcutaneous ectopic osteogenesis models were established, the osteogenesis and neurogenesis were detected by hematoxylin-eosin (HE) staining, Masson staining and tissue immunofluorescence assay. Statistical analysis was conducted by independent sample t test. RESULTS: After gene silencing of NLK by siRNA in hBMSCs, neural-related genes, including the class Ⅲ β-tubulin (TUBB3), microtubule association protein-2 (MAP2), soluble protein-100 (S100), nestin (NES), NG2 proteoglycan (NG2) and calcitonin gene-related peptide (CGRP), were increased significantly in NLK-knockdown hBMSCs compared with the negative control group(P < 0.05), and the expression levels of TUBB3 and MAP2 of the NLK silencing group were also increased. Oppositely, after NLK was overexpressed using lentivirus vector transfection technique, TUBB3, MAP2, S100 and NG2 were significantly decreased in NLK-overexpression hBMSCs compared with the empty vector group (P < 0.05), and the expression level of TUBB3 was also decreased. 4 weeks after the subcutaneous ectopic osteogenesis model was established, more mineralized tissues were formed in the β-TCP+ siRNA-NLK transfection hBMSCs group compared with the other three groups, and the expression of BMP2 and S100 was higher in the β-TCP+ siRNA-NLK transfection hBMSCs group than in the other groups. CONCLUSION Gene silencing of NLK by siRNA promoted the ability of neural differentiation of hBMSCs in vitro and promoted neuralized tissue engineered bone formation in subcutaneous ectopic osteogenic models in vivo in nude mice.
Bone Regeneration/genetics* ; Animals ; Mesenchymal Stem Cells/cytology* ; Humans ; RNA, Small Interfering/genetics* ; Tissue Engineering/methods* ; Cell Differentiation ; Mice, Nude ; Gene Silencing ; Mice ; Male ; Protein Serine-Threonine Kinases/genetics* ; Intracellular Signaling Peptides and Proteins/genetics* ; Transfection ; Cells, Cultured ; Lentivirus/genetics*

Objective: To investigate the status of overweight and obesity among primary and middle school students in Yuyao City, Zhejiang Province, so as to provide the basis for formulating weight management strategies for students. Methods: A census was conducted to investigate 72 primary schools and 23 middle schools students in Yuyao City in 2024, and data such as gender and age were collected through questionnaire surveys. Height and weight were measured to calculate the body mass index (BMI). Based on the BMI thresholds for different gender and age groups, overweight and obesity were determined. Results: Totally 75 082 individuals were surveyed, including 40 435 boys and 34 647 girls. Among them, 55 172 were primary school students and 19 910 were middle school students. A total of 8 677 overweight and 7 042 obese individuals were identified, with detection rates of 11.56% and 9.38%, respectively. The detection rates of overweight and obesity in boys were higher than in girls (14.01% vs. 8.69%, 11.58% vs. 6.81%, both P<0.05). The detection rate of obesity among students in rural schools was higher than students in urban schools (10.12% vs. 8.18%, P<0.05). The detection rates of overweight and obesity of students with non-local household registration were higher than students with local household registration (12.04% vs. 10.77%, 10.44% vs. 7.64%, both P<0.05). The detection rate of overweight showed an upward trend with age (P<0.05), while no significant age-related trend was observed for obesity (P>0.05). Conclusions The detection rates of overweight and obesity were notably high among primary and middle school students in Yuyao City. Boys, students in rural schools, students with non-local household registration, and older students are the key population.

Objective lt aims to identify key issues in the current informatization development of internal control over revenue and expenditure in public hospitals.The findings are intended to serve as a reference for deepening this informatization effort.Methods Following the steps and requirements of content analysis method,it involved a semi-quantitative analysis of policies and expert interviews to establish an analytical framework.Two researchers uti-lized NVivo 15 software to analyze the policy and articles.The analysis results were quantified statistically,and key is-sues were summarized.Results lt developed a content analysis framework comprising 5 primary categories and 17 secondary categories.Based on this framework,326 analytical units extracted from 81 articles were categorized and statistically analyzed.Key problems corresponding to each secondary category were summarized.Conclusion To ad-dress five key issues,it proposes a five-dimensional solution including:building a digital foundation to integrate busi-ness systems;establishing a Master Data Management platform to dismantle data silos;developing flexible ap-proval mechanismsfor medical emergencies;deploying an Al risk control hub for precise payment interception,and implementing zero-trust architecture to enhance e-bill anti-counterfeiting-ultimately forming a business-finance-in-tegrated,data-and-Al-driven,proactively secured internal control system covering budget formulation to fund su-pervision.

Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome, with implications for microbial pathogenesis and host defense. Among these, transfer RNA-derived small RNAs (tsRNAs) have garnered attention for their roles in modulating microbial behavior. However, the bacterial factors mediating tsRNA interaction and functionality remain poorly understood. In this study, using RNA affinity pull-down assay in combination with mass spectrometry, we identified a putative membrane-bound protein, annotated as P-type ATPase transporter (PtaT) in Fusobacterium nucleatum (Fn), which binds Fn-targeting tsRNAs in a sequence-specific manner. Through targeted mutagenesis and phenotypic characterization, we showed that in both the Fn type strain and a clinical tumor isolate, deletion of ptaT led to reduced tsRNA intake and enhanced resistance to tsRNA-induced growth inhibition. Global RNA sequencing and label-free Raman spectroscopy revealed the phenotypic differences between Fn wild type and PtaT-deficient mutant, highlighting the functional significance of PtaT in purine and pyrimidine metabolism. Furthermore, AlphaFold 3 prediction provides evidence supporting the specific binding between PtaT and Fn-targeting tsRNA. By uncovering the first RNA-binding protein in Fn implicated in growth modulation through interactions with host-derived small RNAs (sRNAs), our study offers new insights into sRNA-mediated host-pathogen interplay within the context of microbiome-host interactions.
Fusobacterium nucleatum/growth & development* ; RNA-Binding Proteins/genetics* ; Bacterial Proteins/genetics* ; RNA, Bacterial/metabolism* ; Humans ; RNA, Transfer/metabolism*

[Purpose]To analyze the trends of disease burden for lung cancer worldwide and in China from 1990 to 2021.[Methods]Data of the disease burden of lung cancer and population demographics in 1990 and 2021 stratified by sex and age groups for global,five SDI quintiles re-gions,and eight countries including China,Japan,Republic of Korea,United Kingdom,France,United States,Canada,and Australia were extracted from the Global Burden of Disease Study 2021(GBD 2021)database.The age-standardized mortality rate(ASMR)and age-standard-ized disability-adjusted life year rate(ASDR)of lung cancer attributable to 7 level-3 risk factors in China for 1990 and 2021 were also extracted.Counterfactual analysis was used to decompose changes in lung cancer deaths and DALY from 1990 to 2021 into four contributing factors:popu-lation size,population structure,age-standardized incidence or prevalence,and lung cancer case fatality or disease severity.The percentage changes in lung cancer deaths and DALY attributed to these four factors were calculated respectively.[Results]In 2021,there were 934 704 new cases and 814 364 deaths of lung cancer in China.From 1990 to 2021,the incidence,preva-lence,mortality,and DALY rates of lung cancer in China increased faster than those worldwide and in high-middle SDI regions,which was similar to Japan and Republic of Korea.In contrast,the mortality rates of lung cancer decreased in United States and United Kingdom;and the DALY rates of lung cancer decreased in United States,United Kingdom,Canada and Australia.From 1990 to 2021,the age-related lung cancer deaths and DALY in China increased by 193.91％and 146.20％,respectively.The primary contributor to the increase in lung cancer deaths was population aging(102.82％)among men and rising age-standardized incidence(119.00％)among women,while the primary contributor to the increase in DALY was rising age-standardized prevalence for both men(153.12％)and women(218.77％).In 2021,the top three risk factors contributing to lung cancer ASMR and ASDR in China were smoking,particulate matter pollution and occupational carcinogen exposure.Compared with 1990,the ASMR of lung cancer and its proportion at-tributable to particulate matter pollution and low dietary fruits were decreased,while the propor-tions in ASDR of lung cancer attributable to smoking and secondhand smoke increased.[Conclu-sion]Lung cancer is a major public health challenge in China.Compared with worldwide,high-middle SDI regions and certain developed countries,China has experienced faster growth in the incidence,prevalence,mortality and DALY of lung cancer,especially among women.To reduce disease burden,sustained efforts on lung cancer prevention and control are urgently required in China.

Objective To explore the mechanism of Yizhu Wendan Decoction in treating eczema through GEO database combined with network pharmacology and experimental verification.Methods TCMSP,BATMAN-TCM and ETCM databases were used to screen the active components of Yizhu Wendan Decoction.Disease target information related to eczema was collected through GEO database.The drug-component-target network and PPI network were constructed by intersections of active component targets and disease targets.GO and KEGG pathway enrichment analyses were performed using DAVID database.CCK-8 method was used to screen out the optimal intervention concentration of freeze-dried powder of Yizhu Wendan Decoction.HaCaT cells were divided into control group,model group,Yizhu Wendan Decoction low concentration group,Yizhu Wendan Decoction high concentration group,si-IL-17RA group,si-IL-17RA+Yizhu Wendan Decoction low concentration group,si-IL-17RA+Yizhu Wendan Decoction high concentration group,Dexamethasone group,si-IL-17RA+Dexamethasone group.Each group was given relevant intervention.The expressions of chemokines and inflammatory factors were detected by qPCR.EdU and Annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis.Western blot was performed to detect the expressions of proteins related to apoptosis,skin barrier and IL-17 signaling pathway.Results By using databases,180 active components of Yizhu Wendan Decoction were obtained.Combined with GEO database microarrays related to eczema(GSE6012 and GSE57225),8 potential targets of Yizhu Wendan Decoction in the treatment of eczema were obtained.KEGG enrichment pathway mainly involved IL-17 signaling pathway,lipid and atherosclerotic,TNF signaling pathway,fluid shear stress and atherosclerotic,etc.When Yizhu Wendan Decoction freeze-dried powder concentration was 100 μg/mL,cell viability was the strongest.Yizhu Wendan Decoction could significantly inhibit the mRNA expressions of chemokines and inflammatory factors CCL17,CCL22,IL-1β,TNF-α,IL-6,IFN-γ,and increase the mRNA expression of IL-4 in eczema.It promoted the proliferation of HaCaT cells,increased the protein expression of Bcl-2,and reduced the protein expressions of Bad and Cleaved Caspase-3,thus inhibiting HaCaT cells apoptosis;promoted the protein expressions of FLG and LOR,and reduced the expression of MMP9,MMP1,CCL2,FOSL1,IL-17RA proteins in IL-17 signaling pathway.Conclusion Yizhu Wendan Decoction can treat eczema with multiple components,multiple pathways and multiple targets,promote the proliferation of HaCaT cells,inhibit their apoptosis,and restore the skin barrier.Its mechanism may be related to inhibiting the activation of IL-17 signaling pathway.

Aim To explore the potential mechanism of metformin on ATP-binding cassette transport A1(ABCA1)expression.Methods J774A.1 macrophages were treated with metformin and cycloheximide,and ABCA1 expression was determined by Western blot.His-tagged ABCA1 and HA-tagged Ub plasmids were co-transferred into HEK293 cells and stimulated with metformin.Co-immunoprecipitation(Co-IP)was used to test the binding ability of ABCA1 and ubiquitin.Candidate E3 ubiquitin-protein ligases(CE3)of ABCA1 were identified through Co-IP-based pro-teomics.The MIB1 plasmid was constructed and transferred into HEK293 cells,and Western blot was used to determine the effect of metformin and MIB1 on ABCA1 expression.Results Metformin increased the expression of ABCA1 in J774A.1 cells(P<0.01),and inhibited ABCA1 degradation(P<0.05).Metformin disrupted the binding of ABCA1 to ubiquitin(P<0.05).The proteins regulated by metformin in ABCA1 expression were primarily enriched in pathways re-lated to cell development,inflammation and immune defense.Metformin may upregulate ABCA1 protein expression via MIB1(P<0.05).Conclusion Metformin inhibits the degradation of ABCA1 by blocking the ubiquitin-proteasome system(UPS),and MIB1 might act as a candidate E3 ubiquitin-protein ligase(CE3)for ABCA1.

Objective To search,evaluate and summarise the best evidence on non-drug intervention for nipple pain or injury in breastfeeding mothers.Methods Based on the"6S pyramid"evidence model,desktop searches were conducted on databases of UpToDate,Joanna Briggs Institute evidence-based practice database(JBI),National Guideline Clearinghouse(NGC),National Institute for health and Care Excellence(NICE),Guidelines International Network(GIN),Registered Nurses'Association of Ontario(RNAO),American College of Obstetricians and Gynecologists(ACOG),Medlive,Chinese Medical Association(CMA)website,Breastfeeding Medical Association(BMA),CNKI,Wanfang Data,SinoMed,Cochrance Library,PubMed,Embase,Web of Science,and CINAHL for literature in non-drug interventions for nipple pain or injury in breastfeeding women.The literature included clinical guidelines,decisions,recommended practices,evidence summaries,expert consensus,systematic reviews and randomized controlled trials(RCTs).The searched databases spanned from 1st January,2013 to 30th April,2024.Two researchers who were trained in evidence-based nursing independently evaluated the methodological quality,extracted and integrated evidence from eligible literature.Results A total of 15 documents were included,consisting of 5 clinical guidelines,2 clinical decisions,1 expert consensus,6 systematic evaluations and 1 RCT.Thirty-one pieces of evidence were summarised across 4 categories:accurate perinatal assessment,feeding guidance,non-drug intervention and health education.Conclusion The summarised best evidence on non-drug intervention for nipple pain or injury in breastfeeding women provides an evidence-based basis for clinical healthcare professionals.