The neurotoxicity related to immune checkpoint inhibitors (ICI) is rare, and the clinical experience remains limited. This paper reported an elderly male patient with advanced small cell lung cancer (SCLC). Following two circles of chemotherapy combined with immunotherapy (serplulimab, 200 mg), the patient developed fatigue, cognitive impairment, motor apraxia, and ataxia. Based on the examination of blood neurotransmitters and brain magnetic resonance imaging, the patient was diagnosed with ICI-associated encephalitis, and his symptoms improved after hormone treatment.

Digital medical insurance governance， as an important component of digital transformation， improves the efficiency of medical insurance services while facing complex ethical challenges. Taking stakeholder theory as the analytical framework， this paper explored the generative mechanisms， manifestations， and countermeasures of ethical issues in digital medical insurance governance. This study uncovered that accelerated technological iteration and lagging ethical and legal norms， as well as interest conflicts among multiple participants in governance， constituted the major causes of ethical issues. These issues specifically manifested as data privacy leakage， unfair algorithmic decision-making， widening digital divide， ambiguous accountability， and other aspects. To address these issues， this paper put forward systematic countermeasures， such as establishing a dynamic ethical and legal regulatory system， balancing the demands of stakeholders， strengthening technological ethical governance， narrowing the digital divide， and enhancing accountability mechanisms. Through the implementation of multi-dimensional governance strategies， it aimed to balance technological innovation and ethical constraints， advance the inclusiveness， fairness， and sustainable development of digital medical insurance， and provide theoretical reference and practical references for improving China’s digital medical insurance governance system.

Objective:To investigate the changes of the epidemiology and clinical characteristics of human metapneumovirus (hMPV) among children with acute lower respiratory tract infection (ALRTI) before and after the discontinuation of non-pharmaceutical interventions (NPI) during coronavirus disease 2019 epidemic.Methods:This was a retrospective cohort study. Children hospitalized at The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University between January 2021 and December 2023, who were diagnosed with ALRTI by nasopharyngeal secretion testing for respiratory pathogens nucleic acid were enrolled. Clinical and laboratory data were collected. Children admitted between January 1, 2021 and January 7, 2023 were classified as the pre-NPI withdrawal group (abbreviated as pre-withdrawal group), while those admitted from January 8, 2023 afterward were classified as the post-NPI withdrawal group (abbreviated as post-withdrawal group). Nasopharyngeal secretions from the enrolled children were tested for 13 respiratory pathogens using polymerase chain reaction-capillary electrophoresis fragment analysis, and bacterial cultures were also performed. Statistical analyses were performed using the Mann-Whitney U test or chi-square test. Results:A total of 30 855 ALRTI cases were enrolled, with 1 679 of hMPV-positive. In the pre-withdrawal group, there were 861 cases with an age of onset of 2.0(1.0, 3.0) years, and the highest proportion was in the 1 to <3 years age group, accounting for 35.3%(304/861). In the post-withdrawal group, there were 818 cases with an age of onset of 3.0(2.0, 4.0) years, and the highest proportion was in the 3 to <5 years age group, accounting for 39.2%(321/818).The age of onset in the post-withdrawal group was significantly older than that in the pre-withdrawal group ( Z=7.69, P<0.001) .The hMPV detection rate was higher in the pre-withdrawal group than that in post-withdrawal group (5.75%(861/14 984) vs 5.15%(818/15 871); χ2=5.25, P=0.022). In the pre-withdrawal group, the epidemic peaks occurred in winter and spring, with the highest rates in January 2022(25.2%(224/890)) and March 2022 (21.6%(186/860)). In the post-withdrawal group, the epidemic peak shifted to spring and summer, and the detection rate became increased since April 2023(10.8%(136/1 258)). The post-withdrawal group showed lower rates of wheezing, shortness of breath, cyanosis, respiratory support, severe pneumonia, intensive care unit admission, and shorter hospital stays compared to the pre-withdrawal group ( χ2=69.09, 31.63, 12.97, 57.96, 55.73, 5.48 and Z=7.11, respectively, all P< 0.05).In the pre-withdrawal group, 412 cases (47.9%(412/861)) had other pathogens detected, compared to 445 cases (54.4%(445/181)) in the post-withdrawal group, indicating a significantly higher rate of co-infections in the post-withdrawal group ( χ2=7.20, P<0.05). The most commonly detected pathogens in both groups were Mycoplasma pneumoniae (MP), rhinovirus, and Streptococcus pneumoniae. However, the post-withdrawal group showed significantly higher detection rates of MP and influenza virus, but lower bacterial detection rates compared to the pre-withdrawal group ( χ2=39.41, 9.70, 5.63, respectively, all P<0.05). The detection rate of Haemophilus influenzae was 2.1%(17/818) in the post-withdrawal group which lower than that (6.7%(58/861)) in the pre-withdrawal group, and the difference was statistically significant ( χ2=21.32, P<0.001). Conclusions:In 2023, following the withdrawal of NPI, the epidemic peak of hMPV in Wenzhou area is delayed to spring and summer. The age of children with hMPV-associated ALRTI increases, with the majority being 3 to <5 years old. The overall severity of the disease decreases. However, the detection of mixed pathogens increases, with MP being the most common, while bacterial detection decreases.

Aim To study the possibility of luteolin(LUT)as a new drug of traditional Chinese medicine and its improving effect on glucose and lipid metabo-lism and liver oxidative stress injury in db/db mice,and to explore the possible mechanism.Methods AI was used to predict the drug toxicity,evaluate the phys-ical and chemical properties and segment the molecular structure of LUT.Molecular docking was used to verify the binding ability of LUT with Nrf2 and HMOX1;db/m mice were divided into the group C,and db/db mice were randomly divided into the T,L,M,H and P groups.The body weight and FBG changes were ob-served within 12 weeks of intervention.The expres-sions of FBG,HbA1c,Fins,TC,TG,HDL-C,LDL-C were detected.The pathomorphological changes and steatosis of mouse liver were observed by HE and oil red O staining.The expression of MDA,SOD and GSH-Px in liver was detected by Kit.The protein and mRNA expressions of Nrf2 and HMOX1 were detected by immunohistochemistry,WB and qPCR.Results AI algorithm predicted the safety and easy synthesis of LUT.LUT intervention had no significant effect on the body weight of db/db mice.After 12 weeks,compared with the group C,the livers of mice in group T showed disordered structure of hepatic lobules,irregular ar-rangement of hepatocytes,and a large number of fat vacuoles and lipid droplets in the cytoplasm.Compared with the group T,LUT intervention could improve the pathological changes of liver,reduce the expression of FBG,HbA1c,fins,TC,TG,LDL-C,MDA,improve the level of HOMA-IR,and upregulate the activities of SOD and GSH-Px.Molecular docking results showed LUT had strong binding with Nrf2 and HMOX1,and could increase the expression of Nrf2 and HMOX1.Conclu-sion LUT can correct the disorder of glucose and lip-id metabolism in db/db mice,and improve the level of oxidative stress in liver of mice through Nrf2/HMOX1 pathway,which has the development prospect as a new Chinese medicine for the treatment of T2DM.

Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Background and Aims:Totally implantable venous access ports(TIVAP)are widely used in patients requiring long-term intravenous therapy.Traditional butterfly needle puncture fixation methods have limitations,including low success rates,increased pain,and risk of needle-stick injury.This study aimed to design an adjustable puncture protection kit for butterfly needles and evaluate its clinical utility using a simulated device.Methods:A prospective randomized controlled trial was conducted with 70 patients implanted with upper arm ports in the Hematology Department of Xiangya Hospital,Central South University,from January to December 2024.The patients were divided into a study group and a control group,with 35 cases in each,using a randomized block design.The study group underwent puncture with the simulated adjustable protection kit,while the control group used the traditional finger fixation method.Outcomes compared included first-attempt success rate,vertical puncture rate,pain score,puncture time,and complication rate.Results:The baseline characteristics of the two groups were balanced.The study group had significantly higher first-attempt puncture success rate and vertical puncture rate than the control group(94.3%vs.77.1%;91.4%vs.57.1%,both P<0.05).In the experimental group compared with the control group,pain scores were lower(1.80±1.13 vs.2.94±1.33,P<0.05),and puncture time was shorter[(31.31±9.05)s vs.(41.80±23.97)s,P<0.05].There was no significant difference in the incidence of puncture-related complications between the two groups(2.9%vs.14.3%,P>0.05).Conclusion:The simulated adjustable butterfly needle puncture protection kit effectively improves puncture success,enhances efficiency,reduces patient pain,and demonstrates good clinical safety.This innovative design provides a promising solution for reducing needle-stick injury risks and optimizing port puncture procedures,although larger,multicenter,and long-term studies are warranted.

Objective : To explore the impacts and fundamental mechanisms of the PU. 1 inhibitor DB2313 on the immune function in MRL/lpr mice. Methods: A total of thirty MRL/lpr lupus mice were randomly distributed into three separate groups : the model control group , the PU. 1 inhibitor DB2313 treatment group ( administered at a dose of 17 mg/kg) , and the positive drug control Telitacicept (TACI_Ig) group (administered at a dose of 7. 5 mg/ kg) . Furthermore , a group of ten BALB/c mice were assigned as the normal group. The DB2313 administration group was treated with intraperitoneal injections of the drug on three occasions per week , while the TACI_Ig group received subcutaneous injections every second day; both treatment protocols were maintained for a duration of five weeks. Both the control group and the model group were administered intraperitoneal injections of a volume of saline that was equivalent across groups. After the drug was given , mice were sacrificed by dislocation after orbital vein blood collection. The thymus and spleen were aseptically excised , individually weighed , and subsequently utilized to compute the thymus index and spleen index. The relative distribution of T lymphocyte subsets within the spleen was ascertained through flow cytometry. Serum concentrations of anti_nuclear antibodies ( ANA) and antidouble_stranded DNA antibodies were quantified using an enzyme_linked immunosorbent assay (ELISA) . The levels of inflammatory factors interleukin_6 (IL_6) , tumor necrosis factor_α (TNF_α) , interferon_γ(IFN_γ) were meas ured by CBA method. Hematoxylin and eosin ( HE) staining was employed to examine pathological alterations in the spleen. The expression of PU. 1 and IL_9 in spleen tissue was detected using immunohistochemistry. Additionally , the expression level of PU. 1 protein in the spleen tissue was ascertained through Western blot analysis. Results: The administration of DB2313 significantly ameliorated spleen lesions in MRL/lpr mice and decreased the levels of anti_ds_DNA , ANA , TNF_α , IL_6 , and IFN_γ . It also reduced the proportion of total T cells , TFH cells , Th17 cells , and Th9 cells in the mouse spleen , while increasing the proportion of Treg cells. Furthermore , it lowered the level of PU. 1 protein in the spleen. Immunohistochemistry results demonstrated that DB2313 treatment significantly diminished the expression of PU. 1 and IL_9 in spleen tissue. Conclusion The PU. 1 inhibitor DB2313 can improve spleen lesions in MRL/lpr mice and slow the progression of the disease , and its mechanism is related to the regulation of immune cell functions.

Objective : To establish an interleukin-1β (Il-1β) induced inflammatory model of rat articular chondro- cytes (ACs) , and to investigate the relationship between the expression of lysine acetyltransferase 7 (KAT7) under inflammatory stimulation and the senescence of ACs. Methods: Primary ACs were obtained by digestion of rat knee cartilage with collagenase type Ⅱ and identified. The inflammatory model of ACs was induced by IL-1β . KAT7 was over-expressed or knocked down in ACs by adeno-associated virus infection or small interfering RNA transfection , respectively. A negative control group was set up. Transwell assay was used to detect cell migration ability. Senes- cent cells were stained with senescence-associated β-galactosidase (SA-β-Gal) . Western blot ( WB) was used to detect the protein expression levels of KAT7 , collagen type II (Col Ⅱ ) , matrix metalloproteinase 13 (MMP13) , tumor protein p53 (p53) and cyclin-dependent kinase inhibitor 1A (p21) . The cells of negative control group and KAT7 over-expression group were performed for RNA sequencing , and WB was used to verify the related signaling pathways obtained by Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. Results: Compared with the control group , the SA-β-Gal staining was enhanced , the protein expression of Col Ⅱ decreased , the pro- tein expression of MMP13 and p53 increased , the cell migration ability decreased , and the expression of KAT7 also increased in the ACs of rats after IL-1β stimulation. Compared with the negative control group , the SA-β-Gal stai- ning was enhanced , the protein expression of Col Ⅱ decreased , the protein expression of MMP13 , p53 and p21 in- creased , and the cell migration ability decreased in the KAT7 over-expression group. Compared with the negative control group , the SA-β-Gal staining was weakened , the protein expression of Col Ⅱ increased , the protein expres- sion of MMP13 , p53 and p21 decreased , and the cell migration ability was enhanced in the KAT7 knockdown inflammatory model of ACs. KEGG enrichment analysis showed that phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway was activated. Compared with the negative control group , the relative protein ex⁃pression levels of phosphorylated protein kinase B (p⁃AKT)/AKT and phosphorylated mammalian target of rapamy⁃cin (p⁃mTOR)/mTOR in KAT7 over⁃expression group increased. The relative protein expression levels of p ⁃AKT/AKT and p ⁃mTOR/mTOR in KAT7 knockdown cells decreased. Conclusion Rat ACs with high expression of KAT7 exhibit senescence and osteoarthritis phenotype , and the mechanism may be related to the activation of PI3K/AKT/mTOR signaling pathway by KAT7.

Objective To explore the impacts and fundamental mechanisms of the PU.1 inhibitor DB2313 on the immune function in MRL/lpr mice.Methods A total of thirty MRL/lpr lupus mice were randomly distributed into three separate groups:the model control group,the PU.1 inhibitor DB2313 treatment group(administered at a dose of 17 mg/kg),and the positive drug control Telitacicept(TACI-Ig)group(administered at a dose of 7.5 mg/kg).Furthermore,a group of ten BALB/c mice were assigned as the normal group.The DB2313 administration group was treated with intraperitoneal injections of the drug on three occasions per week,while the TACI-Ig group received subcutaneous injections every second day;both treatment protocols were maintained for a duration of five weeks.Both the control group and the model group were administered intraperitoneal injections of a volume of sa-line that was equivalent across groups.After the drug was given,mice were sacrificed by dislocation after orbital vein blood collection.The thymus and spleen were aseptically excised,individually weighed,and subsequently uti-lized to compute the thymus index and spleen index.The relative distribution of T lymphocyte subsets within the spleen was ascertained through flow cytometry.Serum concentrations of anti-nuclear antibodies(ANA)and anti-double-stranded DNA antibodies were quantified using an enzyme-linked immunosorbent assay(ELISA).The lev-els of inflammatory factors interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)were meas-ured by CBA method.Hematoxylin and eosin(HE)staining was employed to examine pathological alterations in the spleen.The expression of PU.1 and IL-9 in spleen tissue was detected using immunohistochemistry.Addition-ally,the expression level of PU.1 protein in the spleen tissue was ascertained through Western blot analysis.Re-sults The administration of DB2313 significantly ameliorated spleen lesions in MRL/lpr mice and decreased the levels of anti-ds-DNA,ANA,TNF-α,IL-6,and IFN-γ.It also reduced the proportion of total T cells,TFH cells,Th17 cells,and Th9 cells in the mouse spleen,while increasing the proportion of Treg cells.Furthermore,it low-ered the level of PU.1 protein in the spleen.Immunohistochemistry results demonstrated that DB2313 treatment sig-nificantly diminished the expression of PU.1 and IL-9 in spleen tissue.Conclusion The PU.1 inhibitor DB2313 can improve spleen lesions in MRL/lpr mice and slow the progression of the disease,and its mechanism is related to the regulation of immune cell functions.

Objective To explore the role of miR-429 in synovial mesenchymal stem cell-derived exosomes(SMSC-Exos)in repairing cartilage damage in temporomandibular joint osteoarthritis(TMJ OA)by extracting SMSC-Exos from human synovial tissue and screening differentially expressed microRNA(miRNA)through transcriptome sequencing.Methods Human synovial tissues were obtained from 6 patients who underwent surgery at the First Medical Center of the Chinese PLA General Hospital from June to December 2023,including 3 patients with osteoarthritis(OA group)and 3 control patients(control group),all of whom were male.SMSC-Exos were extracted from the synovial tissues for miRNA sequencing and differential expression analysis.Further,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on the target genes of differentially expressed miRNA to identify key functional miRNA and construct miRNA-target gene regulatory networks and protein-protein interaction(PPI)networks of target genes.An in vitro model of rabbit condylar cartilage cell inflammatory microenvironment induced by interleukin-1β(IL-1β)was established,with the control group cultured in DMEM/F12 basic medium and the inflammation-induced group cultured in DMEM/F12 basic medium containing 10 ng/ml IL-1β.RT-qPCR was used to detect the effects of overexpressed target miRNA on the mRNA expression levels of cartilage phenotype factors such as type Ⅱ collagen α1 chain(Col2a1),aggrecan(Acan),as well as inflammatory factors including a disintegrin and metalloproteinase with thrombospondin motifs 5(Adamts5)and cyclooxygenase-2(Cox-2).Results(1)SMSC-Exos were successfully isolated,cultured,and identified.(2)miRNA sequencing of SMSC-Exos from OA and control groups revealed 16 differentially expressed miRNAs(|log2FC|>2,P<0.05).Compared with control group,7 miRNAs were up-regulated and 9 were down-regulated in OA group.GO and KEGG analysis indicated that the target genes of miR-429 were mainly involved in development process,anatomical structure development,system development,cell development and differentiation,and were enriched in inflammation-related pathways such as mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt).(3)Functional validation of miR-429 in the rabbit condylar cartilage cell inflammatory model showed that overexpression of miR-429 increased the mRNA expression levels of Col2a1 and Acan(P<0.05)and decreased the mRNA expression levels of Adamts5 and Cox-2(P<0.05)in the inflammation-induced group.Conclusions miRNA sequencing of SMSC-Exos isolated and identified from human synovial tissues reveals a specific miRNA expression profile in OA patients,with miR-429 significantly down-regulated.Functional validation demonstrates that overexpression of miR-429 has reparative and anti-inflammatory effects on condylar cartilage cells in an inflammatory microenvironment.