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.

OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. （TFCTL） on hepatic stellate cell （HSC） activation based on the microRNA （miRNA）-204/NUAK family SNF1-like kinase 1 （NUAK1）/Hippo signaling axis， thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group， model group， TFCTL low-concentration group （20 μg/mL）， TFCTL medium-concentration group （40 μg/mL）， and TFCTL high-concentration group （60 μg/mL）. Except for control group， the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells， followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed， the expression levels of α-smooth muscle actin （α-SMA）， type Ⅰ collagen （Collagen Ⅰ） and proteins associated with the Hippo/Yes-associated protein （YAP） pathway [YAP， large tumor suppressor kinase 1 （LATS1）， and mammalian STE20-like kinase 1 （MST1）] were detected. Additionally， cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL， the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 （except for the TFCTL high-concentration group） and LATS1 were significantly increased （P＜0.05）， while the protein expressions of α-SMA， CollagenⅠ， and YAP （except for the TFCTL medium-concentration group） were significantly decreased （P＜0.05）. Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics， the mRNA it’s protein expressions of α-SMA， CollagenⅠ， NUAK1， and YAP in HSC-T6 cells were significantly decreased （P＜0.05）， while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased （P＜0.05）. Conversely， the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204， thereby down- regulating the expressions of NUAK1， inactivating the Hippo/YAP pathway， which in turn suppresses the activation of HSC and promotes their apoptosis.

Objective: To analyze the changes in classroom lighting environment of schools in Suzhou and their impact on refractive progression among primary and secondary school students, so as to provide the basis for accurate provention and control of myopia. Methods: A baseline investigation was conducted in October 2022 by using a stratified cluster random sampling method to recruit primary and secondary school students from Suzhou. A follow up visit was performed in October 2023. A total of 12 302 students and 360 classrooms that participated in both surveys were included analysis. The visual acuity progression over one year and classroom lighting conditions were assessed. Group comparisons were performed by using the Wilcoxon or Kruskal-Wallis rank-sum, and Chi-square tests. Multivariate Logistic regression was employed to identify the major factors influencing refractive changes. Results: The compliance rate of average illuminance on classroom blackboard surface increased from 72.22% to 75.28%, while the compliance rate of average illuminance on desks decreased from 89.44% to 87.22%, the overall myopia rate among students rose from 59.63% to 66.99% from 2022 to 2023. The average annual progression of equivalent spherical power(SE) in the right eye of students was -0.25(-0.75,0.06)D. Significant statistical differences were observed in the annual mean changes across different school levels, regions, baseline refractive statuses, and classroom lighting environment change groups ( Z/H =316.59, -8.27, 38.80 , 51.01, all P <0.05). Multivariate Logistic regression analysis showed that pre myopia, low myopia, junior high school, senior high school, vocational high school, and improved classroom lighting environment were protective factors of reducing the risk of rapid progression in refractive error ( OR =0.58, 0.69, 0.81, 0.50, 0.28, 0.82, all P <0.05). Conversely, female students and rural students had higher risks of rapid myopia progression ( OR =1.09, 1.42, both P <0.05). Conclusions Over one year follow up, the complance rate of classroom lighting indicators in Suzhou remaines stable, while students refractive status shows a trend toward myopia. Improving classroom lighting environment can reduce the risk of rapid myopia progression.

Cataract is the world's leading cause of blindness, and surgery is the most effective treatment for cataract. With the development of femtosecond laser technology and ophthalmic surgical equipment, the application of femtosecond laser systems in cataract surgery is becoming increasingly widespread. It can be used in cataract surgery for corneal incisions, anterior capsulotomy, lens fragmentation, arcuate incisions and other key operations. Compared to traditional surgery, femtosecond laser assisted cataract surgery(FLACS)offers significant advantages in precision, safety and postoperative visual outcomes. Its clinical benefits have garnered growing recognition among ophthalmologists. However, the key technologies and high-precision equipment for FLACS remain predominantly controlled by Western countries. In China, the research in this field began later. This article reviews the technological advancements in FLACS, with a focus on femtosecond laser technology, optical coherence tomography(OCT), artificial intelligence, and clinical application progress. The objective is to provide theoretical foundations and practical insights for the development of ophthalmic medical technology in China.

Objective To explore the mechanism of excessive mechanical stress regulated ferroptosis induced by Piezo1 channel in mouse chondrocytes.Methods The experiment was performed on mouse ATDC5 chondrocytes.siRNA-Piezo1 interference plasmid and Piezo1 overexpression plasmid were used to transfect chondrocytes,and mechanical stress stimulation was given.The control group,the mechanical stress stimulation group(MS group),the MS+siRNA-Piezo1 group(MS+sh group)and the MS+Piezo1 overexpression group(MS+OV group)were constructed,respectively.The cell viability,Fe2+,ROS levels,the expression of ferroptosis-related proteins SLC7A11 and GPX4,and the expression of Collagen Ⅱ,MMP-13,Aggrecan and p53 proteins were detected in each group.Results Compared with the control group,the cell viability was decreased in the MS group(P＜0.05).Levels of Fe2+,reactive oxygen species(ROS)and malondialdehyde(MDA)were increased(P＜0.05).Levels of reduced glutathione(GSH)and superoxide dismutase(SOD)were decreased(P＜0.05),and the mitochondrial ridge was decreased detected by transmission electron microscopy.Protein levels of SLC7A11,GPX4,Collagen Ⅱ and Aggrecan were decreased(P＜0.05),while protein levels of p53 and MMP-13 were increased(P＜0.05).Compared with the MS group,Fe2+,ROS and MDA levels were decreased in the MS+sh group(P＜0.05),GSH and SOD levels were increased(P＜0.05),and protein levels of SLC7A11,Collagen Ⅱ,GPX4 and Aggrecan were increased(P＜0.05).The protein levels of MMP-13 and p53 were decreased(P＜0.05).Compared with the MS group,cell viability was decreased(P＜0.05),Fe2+,ROS and MDA levels were increased(P＜0.05),GSH and SOD levels were decreased(P＜0.05),and protein levels of SLC7A11,Collagen Ⅱ,GPX4 and Aggrecan were decreased in the MS+OV group(P＜0.05).Levels of MMP-13 and p53 protein were increased(P＜0.05).Conclusion Excessive mechanical stress can induce chondrocyte ferroptosis and promote extracellular matrix degradation via Piezo1 channel protein.

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 develop nanobodies with broad-spectrum reactivity,specificity,and high sensitivity that can be used for detecting multiple subtypes of influenza A virus,and to establish a nanobody-based enzyme-linked immunosorbent assay(ELISA)method.Methods Gene sequences of twelve nanobodies against influenza A virus were retrieved from the National Center for Biotechnology Information(NCBI)and nanobody databases.The nanoantibodies were prepared using molecular biological techniques including gene synthesis and recombinant expression.The binding activity,specificity,sensitivity,and affinity of these nanobodies were determined by ELISA screening and Gator affinity analysis.A double-antibody sandwich ELISA assay was established by combining the selected nanobody with a traditional mouse monoclonal antibody.Results Twelve nanobodies were expressed and purified.Two nanobodies capable of binding to multiple subtypes of influenza virus including H1,H3,H5,H7,and H9 were obtained and designated as VHH54 and KV108.Both nanobodies showed no cross-reactivity with other respiratory virus antigens.Furthermore,the KV108 nanobody exhibited the highest binding affinity,with a dissociation constant of 5.94×10-9mol/L for the influenza virus nucleoprotein(NP),and the lowest detection concentration for the NP antigen reached 0.00064 μg/mL.The double-antibody sandwich ELISA,using a combination of KV108 and a mouse monoclonal antibody,could sensitively detect the five common subtypes of influenza A virus(H1N1,H3N2,H5N1,H7N9,and H9N2).The lowest detection limit reached 110-403 PFU/mL,which was higher than that of the commercial colloidal gold kitfor influenza virus detection.Conclusion This study has identified a nanobody KV108,which is capable of binding to multiple subtypes of influenza virus,and established a nanobody-based ELISA method that can detect multiple subtypes of influenza A virus.This study can facilitate the development of nanobody-based influenza detection technologies.

Objective To establish human colorectal cancer(CRC)organoids and to evaluate the efficacy of chem-otherapy drugs.Methods Patient-derived CRC cells were cultured to form organoids.The CRC organoids and origi-nal tissues were stained with molecular markers of CRC immunohishtochemically.CRC organoids were used to test drug sensitivity and different concentrations of chemotherapy drugs 5-fluorouracil,oxaliplatin and irinotecan were given respectively;Organoid activity before and after drug treatment was measured by 3D cell viability assay.Results The patient-derived organoids(PDO)from 5 CRC tissues were successfully established.The expression of CK20,Ki67 and Villin proteins was similar in organoids and in original tumor.The organoids retained histologcial features similar to those of the original tumors.Different PDO showed differential sensitivity to different chemothera-py drugs.Conclusions CRC-PDO can dispaly their different sensitivities to different chemotherapy drugs,and could provide valuble reference for personalized treatment for CRC patients.

Objective To systematically evaluate the psychometric properties of specific self-reported outcome assessment tools for patients undergoing Endoscopic Transnasal Transsphenoidal Surgery(ETTS),with the aim of pro-viding clinical healthcare professionals with evidence to guide the selection of appropriate assessment tools.Methods A systematic search was performed across multiple databases,including the North American Skull Base Society,PubMed,Embase,CINAHL,Web of Science,Cochrane Library,CNKI,VIP database,Wanfang database,and the China Biomedical Literature Database,for studies published from the inception of each database to January 9,2025,that are related to ETTS-specific symptom assessment tools.Totally 2 independent researchers screened the literature and extracted relevant data.The consensus-based health measurement tool selection bias risk checklist and quality standards were used to evaluate the methodological quality and psychometric properties of the included studies,which helped to form the development of recommendations.Results A total of 11 articles were included,covering 8 specific self-reported outcome assessment tools for patients undergoing endonasal transsphenoidal pituitary tumor resection.Of these,the Pituitary Outcome Score and the Leiden Bother and Needs Questionnaire for patients with pituitary disease 2 tools were rated as Grade A recommendations,and 6 tools were rated as Grade B recommendations.Conclusion POS and LBNQ-Pituitary are recommended due to their relatively strong overall methodological quality and psychometric properties.However,further validation with larger sample sizes is necessary.

Objective:To investigate the influence of prealbumin on cerebral infarction severity, hemorrhage transformation and 1-year prognosis.Methods:A retrospective study was conducted to select 752 patients with cerebral infarction who were treated in Beijing Tiantan Hospital,Capital Medical University from December 2018 to December 2019 as the study objects. Personal information and laboratory indicators of the patients were collected including prealbumin, hemoglobin, white blood cell count, etc.Patients were divided into group B1 (<238 mg/L) and group B2 (≥238 mg/L) based on median prealbumin. By inquiry patient's case, NIHSS score (<16 was classified as mild, ≥16 as moderate and severe)and cerebral infarction volume (<20 cm 3 as small infarct, >20 cm 3 as large infarct) were recorded to evaluate the severity of the disease, and whether hemorrhage transformation occurred during hospitalization was recorded. Patients were followed up 1 year after discharge, and prognostic information of patients was recorded, including neurological function recovery (mRS score <3 was classified as good recovery, ≥3 as poor recovery),all-cause case fatality rate, and recurrence of cardio-cerebrovascular events. Normally distributed measurement data were expressed as xˉ±s, non-normally distributed measurement data were expressed as median and quartiles[ M( Q1, Q3)], categorical variable were expressed as ratio and percent(%). Comparison between groups of measurement data were performed by independent sample t test and Mann-Whitney U test. Chi-square test were used on comparison between groups of categorical variable. Single-factor comparison, Spearman correlation analysis and multiple Logistic regression were used to analyze the correlation between prealbumin and other laboratory indicators, cerebral infarction severity, hemorrhage transformation and 1-year prognosis, respectively. Results:The NIHSS score and infarct volume of patients in group B1 were 5(2,10) and 3.18(0.72,18.00) cm 3, and those in group B2 were 3(2,7) and 2.0(0.5,10.0) cm 3, respectively, which were higher in group B1 than in group B2, the differences were statistically significant ( Z=3.85, P<0.001, Z=2.81, P=0.005). The proportion of mRS Score ≥3 in group B1 was 28.8%(107/371), and the all-cause case fatality rate was 7.5%(28/371), both higher than 20.5%(78/381) and 3.1%(12/381) in group B2, with statistical significance ( χ2=7.10, P=0.008, χ2=7.22, P=0.007). Hemorrhage transformation and recurrence of cardio-cerebrovascular events were 13.5%(50/371) and 11.6%(43/371) in group B1 and 9.2% (35/381) and 8.7%(33/381) in group B2, respectively, with no significant difference between the two groups ( χ2=3.45, P=0.063, χ2=1.78, P=0.183). Multivariate logistic regression analysis showed that, after adjusted for potential confounding factors, prealbumin was protective factor of NIHSS ( OR and 95% CI: 0.990(0.984-0.997), P=0.035), poor neurological recovery(mRS≥3) ( OR and 95% CI:0.992(0.988-0.997), P<0.001) and all-cause case fatality rate ( OR and 95% CI:0.991(0.983-0.999), while prealbumin had no significant influence on cardiocerebrovascular recurrence events ( OR and 95% CI: 0.999(0.993-1.005), P=0.729). Conclusion:Prealbumin is significantly associated with the severity of cerebral infarction and poor prognosis 1 year after discharge, and low prealbumin was an independent risk factor for NIHSS score(≥16), poor neurological recovery (mRS≥3) and all-cause case fatality rate.