OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets， followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established ， and the zebrafish were treated with L. ruthenicum Murr. extract （LRME） at concentrations of 0.1， 0.2 and 0.4 mg/mL， respectively； 24 h later， behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 （IL-6）， IL-1β， IL-10， tumor necrosis factor-α （TNF-α）]， oxidative stress markers [superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， catalase （CAT）]， and neurotransmitters [5- hydroxytryptamine （5-HT）， γ-aminobutyric acid （GABA）， glutamic acid （Glu）， dopamine （DA）， and norepinephrine （NE）] were measured. The protein expression levels of protein kinase B1 （AKT1）， phosphorylated AKT1 （p-AKT1）， epidermal growth factor receptor （EGFR）， B-cell lymphoma 2 （Bcl-2）， sarcoma proto-oncogene，non-receptor tyrosine kinase （SRC）， and heat shock protein 90α family class A member 1 （HSP90AA1） in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these， apigenin， naringenin and others were recognized as core active compounds， while AKT1， EGFR and others served as key targets； EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-， medium-， and high-dose groups were 54.60%， 69.03% and 77.97%， 开发。E-mail：hjp_yft@163.com respectively， while the inhibition ratios of locomotor distance were 0.57， 0.83 and 0.95， respectively. Compared with the model group， the number of resting counts， resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups （P＜0.05）. Neuronal damage in the brain was alleviated. Additionally， the levels of IL-6， IL-1β， TNF-α， MDA， Glu， DA and NE， as well as the protein expression levels of AKT1， p-AKT1， EGFR， SRC and HSP90AA1， were markedly reduced （P＜0.05）， while the levels of IL-10， SOD， GSH-Px， CAT， 5-HT and GABA， as well as Bcl-2 protein expression， were significantly elevated （P＜0.05）. CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects， and its specific mechanism may be related to the regulation of inflammatory responses， oxidative stress， neurotransmitter balance， and the EGFR tyrosine kinase inhibitor resistance signaling pathway.

The endoplasmic reticulum quality control(ERQC)system is a core mechanism for maintaining cellular homeostasis,which primarily mediates the degradation of misfolded proteins in the endoplasmic reticulum(ER)through the ER-associated degradation(ERAD)and ER autophagy(ER-phagy)pathways.Tripartite motif 13(TRIM13)is a protein located on the ER membrane,which plays a critical role in ERAD via its E3 ubiquitin ligase activity.TRIM13 also acts as a non-classical ER-phagy receptor to mediate the occurrence of ER-phagy.TRIM13 has recently received extensive attention in the field of ERQC.Here we review the structure and function of TRIM 13 and the mechanisms by which it contributes to ERQC,and summarize its abnormal expression and regulatory role in diseases,with the aim of providing new strategies for the treatment of related diseases.

Objective To develop a virtual reality rehabilitation support system and evaluate its effects on improving discharge readiness in caregivers of laryngectomy patients.Methods The system is equipped with a virtual reality-based home nursing skills guidance module for post-laryngectomy patients and an evidence-based artificial intelligence Q&A module.A convenience sampling method was adopted to select 64 caregivers of patients who underwent laryngectomy for the first time in the otorhinolaryngology head and neck surgery department of a tertiary A hospital in Beijing from July 2024 to January 2025 for a systematic study on application effects.To avoid cross-contamination between an experimental group and a control group,ward-based grouping was applied.A coin toss was used to assign Ward 1 to an experimental group and Ward 2 to a control group,with 32 cases in each group.The experimental group received discharge guidance based on the virtual reality technology,while the control group was provided with conventional verbal and written discharge instructions.The readiness for hospital discharge and the quality of discharge teaching scores of caregivers were compared between the 2 groups after the intervention.Results No sample detachment.After the intervention,the patient caregivers in the experimental group scored significantly higher than those in the control group in terms of discharge readiness and quality of discharge guidance,with a statistically significant difference(P＜0.001).Conclusion This study developed the virtual reality rehabilitation system for laryngeal cancer patients demonstrated good feasibility and effectiveness.The application of this system for discharge education significantly improved discharge readiness of caregivers and quality of discharge guidance for post-laryngectomy patients.It also helped with the optimization of health education models,enhancement of nursing resource utilization efficiency,and improvement of transitional care.

BACKGROUND:Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy.Therefore,exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.OBJECTIVE:To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques,in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.METHODS:Two datasets of pulmonary fibrosis,GSE24206 and GSE110147,were downloaded from the Gene Expression Omnibus(GEO)database(a public database developed and maintained by the U.S.National Center for Biotechnology Information to store and share bioinformatics data),and the gene expression matrices of these two datasets were normalized by using the"limma"package in R software.The autophagy-related genes were extracted from GeneCards database(a database created by the U.S.National Center for Biotechnology Information,which automatically integrates gene-centric data from about 200 Web sources,including genomic,transcriptomic,proteomic,genetic,clinical,and functional information).Differential gene analysis was performed on the pulmonary fibrosis dataset,and the common genes were extracted by cross-comparing the differential genes with the autophagy genes,so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis.The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes.Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning,and core genes were subjected to the enrichment analysis.Diagnostic models were constructed from the identified core genes.Calibration curves were used to assess the predictive ability of the line graph model.An external dataset,GSE21369,was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy,as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database.Finally,human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment,and the relative expression of genes in the model cells was verified using qRT-PCR.RESULTS AND CONCLUSION:(1)A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained.Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue,collagen metabolism,collagen pro-fibroblasts,and growth factor binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions.(2)Immunoinfiltration analysis revealed that the expression of activated memory CD4+T cells,M0 macrophages,and resting dendritic cells was significantly elevated in the pulmonary fibrosis group(P＜0.05),showing a strong correlation.(3)Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified:COL1A2 and IL6.The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately,and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance.COL1A2 and IL6 were related to the cell-cycle pathway,mitogen-activated protein kinase signaling pathway,Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions.A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes,and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi.COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis.To conclude,COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis,but its specificity to pulmonary fibrosis needs to be further investigated.

Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.

Objective To develop a virtual reality rehabilitation support system and evaluate its effects on improving discharge readiness in caregivers of laryngectomy patients.Methods The system is equipped with a virtual reality-based home nursing skills guidance module for post-laryngectomy patients and an evidence-based artificial intelligence Q&A module.A convenience sampling method was adopted to select 64 caregivers of patients who underwent laryngectomy for the first time in the otorhinolaryngology head and neck surgery department of a tertiary A hospital in Beijing from July 2024 to January 2025 for a systematic study on application effects.To avoid cross-contamination between an experimental group and a control group,ward-based grouping was applied.A coin toss was used to assign Ward 1 to an experimental group and Ward 2 to a control group,with 32 cases in each group.The experimental group received discharge guidance based on the virtual reality technology,while the control group was provided with conventional verbal and written discharge instructions.The readiness for hospital discharge and the quality of discharge teaching scores of caregivers were compared between the 2 groups after the intervention.Results No sample detachment.After the intervention,the patient caregivers in the experimental group scored significantly higher than those in the control group in terms of discharge readiness and quality of discharge guidance,with a statistically significant difference(P＜0.001).Conclusion This study developed the virtual reality rehabilitation system for laryngeal cancer patients demonstrated good feasibility and effectiveness.The application of this system for discharge education significantly improved discharge readiness of caregivers and quality of discharge guidance for post-laryngectomy patients.It also helped with the optimization of health education models,enhancement of nursing resource utilization efficiency,and improvement of transitional care.

The endoplasmic reticulum quality control(ERQC)system is a core mechanism for maintaining cellular homeostasis,which primarily mediates the degradation of misfolded proteins in the endoplasmic reticulum(ER)through the ER-associated degradation(ERAD)and ER autophagy(ER-phagy)pathways.Tripartite motif 13(TRIM13)is a protein located on the ER membrane,which plays a critical role in ERAD via its E3 ubiquitin ligase activity.TRIM13 also acts as a non-classical ER-phagy receptor to mediate the occurrence of ER-phagy.TRIM13 has recently received extensive attention in the field of ERQC.Here we review the structure and function of TRIM 13 and the mechanisms by which it contributes to ERQC,and summarize its abnormal expression and regulatory role in diseases,with the aim of providing new strategies for the treatment of related diseases.

OBJECTIVE To analyze the reliability of Xpert MTB/RIF(referred to as Xpert)in detecting rifampicin resistance under varying bacterial loads of Mycobacterium tuberculosis(MTB)so as to reference for clinical diag-nosis and treatment.METHODS A total of 3 050 mycobacterial culture-positive test results from Jan.2016 to Dec.2023 at the First Affiliated Hospital of Xiamen University were selected.After excluding 220 non-tubercu-lous mycobacteria cases,2 830 culture-positive specimens underwent Xpert testing,with 2 412 positive and 418 negative cases.Based on MTB bacterial load,specimens were divided into four treatment groups-high,medi-um,low and very low-with 552,916,656 and 288 cases,respectively.Solid phenotypic drug susceptibility testing(DST)was performed on 1 801 MTB strains that were both culture-positive and Xpert-positive.RESULTS The positive rates of rifampicin in the high,medium,low and very-low treatment groups were 10.33％(57/552),11.14％(102/916),8.54％(56/656)and 8.68％(25/288),respectively.Using the solid phenotypic drug sus-ceptibility testing results as the reference standard,the specificity and negative predictive value of Xpert for detec-ting rifampicin resistance in all four bacterial load groups exceeded 96％,with no statistically significant differ-ences observed.No significant difference in sensitivity was noted among the groups(P=0.053),though the high bacterial load group exhibited relatively lower sensitivity at 84.62％(44/52),while the very-low bacterial load group showed the lowest sensitivity at 76.00％(19/25),which was lower than the medium bacterial load group's 94.05％(79/84).The positive predictive values(PPV)across all groups were generally low but without statisti-cally significant differences(P=0.239),with the high and very-low bacterial load groups registering PPVs of 77.19％(44/57)and 76.00％(19/25),respectively.The concordance rates of Xpert and solid phenotypic drug susceptibility testing were consistently high among all four bacterial load groups,including 94.95％(395/416),97.51％(666/683),96.75％(477/493)and 94.26％(197/209),respectively,with no significant difference(P=0.052).The Kappa values from high to very-low bacterial load groups were 0.778,0.889,0.831 and 0.727,re-spectively.Among the strains with discordant results between Xpert and phenotypic drug susceptibility testing,the rifampicin result consistency rates between Xpert and gene chip were 81.82％(9/11),100.00％(9/9),100.00％(2/2)and 25.00％(1/4),respectively for the high to very-low bacterial load groups.CONCLUSIONS When the bacterial load of MTB is very low,the Xpert assay demonstrates a higher probability of both false-posi-tive and false-negative results for rifampicin resistance,followed by high bacterial loads.The most reliable detec-tion of rifampicin resistance occurs with medium bacterial loads.

Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.

OBJECTIVE To analyze the reliability of Xpert MTB/RIF(referred to as Xpert)in detecting rifampicin resistance under varying bacterial loads of Mycobacterium tuberculosis(MTB)so as to reference for clinical diag-nosis and treatment.METHODS A total of 3 050 mycobacterial culture-positive test results from Jan.2016 to Dec.2023 at the First Affiliated Hospital of Xiamen University were selected.After excluding 220 non-tubercu-lous mycobacteria cases,2 830 culture-positive specimens underwent Xpert testing,with 2 412 positive and 418 negative cases.Based on MTB bacterial load,specimens were divided into four treatment groups-high,medi-um,low and very low-with 552,916,656 and 288 cases,respectively.Solid phenotypic drug susceptibility testing(DST)was performed on 1 801 MTB strains that were both culture-positive and Xpert-positive.RESULTS The positive rates of rifampicin in the high,medium,low and very-low treatment groups were 10.33％(57/552),11.14％(102/916),8.54％(56/656)and 8.68％(25/288),respectively.Using the solid phenotypic drug sus-ceptibility testing results as the reference standard,the specificity and negative predictive value of Xpert for detec-ting rifampicin resistance in all four bacterial load groups exceeded 96％,with no statistically significant differ-ences observed.No significant difference in sensitivity was noted among the groups(P=0.053),though the high bacterial load group exhibited relatively lower sensitivity at 84.62％(44/52),while the very-low bacterial load group showed the lowest sensitivity at 76.00％(19/25),which was lower than the medium bacterial load group's 94.05％(79/84).The positive predictive values(PPV)across all groups were generally low but without statisti-cally significant differences(P=0.239),with the high and very-low bacterial load groups registering PPVs of 77.19％(44/57)and 76.00％(19/25),respectively.The concordance rates of Xpert and solid phenotypic drug susceptibility testing were consistently high among all four bacterial load groups,including 94.95％(395/416),97.51％(666/683),96.75％(477/493)and 94.26％(197/209),respectively,with no significant difference(P=0.052).The Kappa values from high to very-low bacterial load groups were 0.778,0.889,0.831 and 0.727,re-spectively.Among the strains with discordant results between Xpert and phenotypic drug susceptibility testing,the rifampicin result consistency rates between Xpert and gene chip were 81.82％(9/11),100.00％(9/9),100.00％(2/2)and 25.00％(1/4),respectively for the high to very-low bacterial load groups.CONCLUSIONS When the bacterial load of MTB is very low,the Xpert assay demonstrates a higher probability of both false-posi-tive and false-negative results for rifampicin resistance,followed by high bacterial loads.The most reliable detec-tion of rifampicin resistance occurs with medium bacterial loads.