Objective To investigate the effect of diosgenin on the proliferation and apoptosis of breast cancer cells and its potential molecular mechanism. Methods The breast cancer cell line MCF-7 was treated with low, medium, and high doses of diosgenin, and cell proliferation was detected through the MMT method. Flow cytometry was used to detect cell apoptosis. Nuclear-cytoplasmic-protein separation method was applied to detect the subcellular localization of death associated protein (DAXX). qRT-PCR and Western blot were used to detect the expressions of DAXX and c-Jun N-terminal kinase pathway (JNK)-related proteins. Results Diosgenin considerably inhibited the proliferation of MCF-7 cells and promoted cell apoptosis in a concentration-dependent manner. Diosgenin can promote the movement of DAXX from nucleus into the cytoplasm. Diosgenin upregulated the expression of cell surface death receptor (Fas), increased the phosphorylation levels of JNK and mitogen activated protein kinase (p38), and activated the JNK/p38 signaling pathway with concentration dependence. Conclusion Diosgenin inhibits the proliferation and promotes the apoptosis of the breast cancer cell line MCF-7, whose mechanism may be related to the regulation of DAXX subcellular localization and the activation of JNK/p38 signaling pathway.

Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a promising target for sensitizing solid tumors to immune checkpoint blockades. However, the highly polar active sites of PTPN2 hinder drug discovery efforts. Leveraging small interfering RNA (siRNA) technology, we developed a novel glutathione-responsive nano-platform HPssPT (HA/PEIss@siPtpn2) to silence PTPN2 and enhance immunotherapy efficacy in hepatocellular carcinoma (HCC). HPssPT showed potent transfection and favorable safety profiles. PTPN2 deficiency induced by HPssPT amplified the interferon γ signaling in HCC cells by increasing the phosphorylation of Janus-activated kinase 1 and signal transducer and activator of transcription 1, resulting in enhanced antigen presentation and T cell activation. The nano-platform was also able to promote the M1-like polarization of macrophages in vitro. The unique tropism of HPssPT towards tumor-associated macrophages, facilitated by hyaluronic acid coating and CD44 receptor targeting, allowed for simultaneous reprogramming of both tumor cells and tumor-associated macrophages, thereby synergistically reshaping tumor microenvironment to an immunostimulatory state. In HCC, colorectal cancer, and melanoma animal models, HPssPT monotherapy provoked robust antitumor immunity, thereby sensitizing tumors to PD-1 blockade, which provided new inspiration for siRNA-based drug discovery and tumor immunotherapy.

Objective To explore the predictive value of early variability of levels of multiple inflammatory factors for the prognosis of children with Mycoplasma pneumoniae pneumonia(MPP).Methods A total of 253 MPP children admitted to the hospital from May 2022 to October 2024 were selected as the research sub-jects.Blood samples were collected from all patients at admission and up to 72 h[T0(0-6 h at admission),T1(24 h after admission),T2(48 h after admission),T3(72 h after admission)].Serum inflammatory fac-tors,including C-reactive protein(CRP),interleukin(IL)-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-17A,IL-1β,γ-in-terferon(IFN-γ),and tumor necrosis factor-α(TNF-α)were measured,and variability was calculated(T1=[(T1-T0)/T0]× 100％,△T2=[(T2-T0)/T0]× 100％,△T3=[(T3-T0)/T0]× 100％).MPP patients were divided into the good prognosis group and poor prognosis according to the prognosis condition.The ser-um inflammatory factor levels and their variability at different time points in the good prognosis group and the poor prognosis group were compared,multivariate Logistic regression was used to analyze the influencing fac-tors of MPP children's prognosis,and receiver operating characteristic(ROC)curve was used to analyze the predictive value of early variability of inflammatory factor levels for MPP children's prognosis.Results After 3 months of follow-up observation,a total of 7 cases were lost to follow-up,and a total of 246 cases were final-ly included for analysis.Among them,58 cases were in the poor prognosis group,and 188 cases were in the good prognosis group.The disease course of the poor prognosis group was longer than that of the good prog-nosis group(P＜0.05),the severity of the disease was more severe than that of the good prognosis group(P＜0.05),and the proportion of affected lung lobes≥2/3 was higher in the poor prognosis group than in the good prognosis group(P＜0.05).IL-6,IL-17A and IL-1β levels at T1-T3 were higher than those at T0(P＜0.0125).The level of IL-6 at T0-T3,IL-1 β level at T2-T3,and IL-17A at T3 in the poor prognosis group were higher than those in the good prognosis group(P＜0.05).IL-6(△T2),IL-17A(△T3),IL-1β(△T2),IL-1β(△T3)in the poor prognosis group were higher than those in the good prognosis group(P＜0.05).The results of the multivariate Logistic regression analysis showed that IL-6(T0),IL-6(T1),IL-6(△T2),IL-1β(△T3),IL-17A(△T3)were the factors influencing the prognosis of children with MPP(P＜0.05).The ROC curve showed that the area under the curve of IL-6(T0),IL-6(T1),IL-6(△T2),IL-1β(△T3),IL-17A(△T3)for predicting the prognosis of children with MPP was all above 0.7.Conclusion The prognosis of MPP patients is related to the phase sensitivity and dynamic remodeling of the inflammatory network.IL-6(T0),IL-6(T1),IL-6(△T2),IL-1β(△T3),IL-17A(△T3)are key prognostic indicators for MPP patients.Dy-namic monitoring of inflammatory factor levels can help intervene in immune imbalances and cytokine storms in a timely manner to improve the prognosis of pediatric patients.

Objective To analyze the correlations of serum levels of brain-derived neurotrophic factor(BDNF),malondialdehyde(MDA),endothelin-1(ET-1)and heat shock protein 70(HSP-70)with cognitive dysfunction in patients with obstructive sleep apnea hypopnea syndrome(OSAHS).Methods A total of 132 OSAHS patients were selected as the study subjects,and another 90 healthy individuals who underwent physical examination during the same period were chosen as control group.According to the severity of the condition,OSAHS patients were divided into mild group(n=43),moderate group(n=50)and severe group(n=39).Serum levels of BDNF,MDA,ET-1 and HSP-70 were compared among different groups.Cognitive function was assessed using the Montreal Cognitive Assessment(MoCA)scale and Mini-Mental State Examination(MMSE).The correlations of cognitive dysfunction,serum levels of BDNF,MDA,ET-1 as well as HSP-70 with disease severity in OSAHS patients were analyzed.Based on MoCA and MMSE scores,OSAHS patients were categorized into cognitive dysfunction group and non-cognitive dysfunction group.Multivariate Logistic regression a-nalysis was used to screen for risk factors of cognitive dysfunction in OSAHS patients.Results The serum BDNF level,MMSE score and MoCA score in the severe group were significantly lower than those in the mild and moderate groups,while serum MDA,ET-1 and HSP-70 levels were signifi-cantly higher(P＜0.05).In the cognitive dysfunction group,the proportion of low education,se-vere disease,body mass index,serum levels of MDA,ET-1 and HSP-70 were significantly higher than those in the non-cognitive dysfunction group,while serum BDNF level was significantly lower(P＜0.05).Serum BDNF level was negatively correlated with cognitive dysfunction,whereas serum MDA,ET-1 as well as HSP-70 levels and disease severity were positively correlated(P＜0.05).Low education,high BMI,high levels of MDA,ET-1 as well as HSP-70 and low BDNF level were risk factors for cognitive dysfunction in OSAHS patients(P＜0.05).Conclusion In OSAHS pa-tients,serum MDA,ET-1 and HSP-70 levels are increased,while serum BDNF level are de-creased.Serum levels of MDA,ET-1,HSP-70 and BDNF are closely related to the occurrence of cognitive dysfunction in OSAHS patients.

Objective To investigate the effects of indirubatin derivative E804 on proliferation and migration of non-small cell lung cancer(NSCLC)A549 cells,and to elucidate the possible mechanism of Nrf2-HO-1/GPX4 pathway.Methods Lung cancer A549 cells were used as the cell model.The proliferation and migration of differ-ent specific inhibitors(Nec-1,CQ,Z-VAD,DFO,Fer-1 and Lip-1)in 0,10 μmol/L E804 and 10 μmol/L E804+groups were observed by MTT and cell scratch assay.The contents of reactive oxygen species(ROS)were de-tected by DCFH-DA fluorescence probe method,the contents of Fe2+were detected by colorimetric method,the contents of reduced glutathione(GSH)were detected by spectrophotometry,and the contents of malondialdehyde(MDA)were detected by micromethod.The expression levels of SLC7A11,Transferrin,GPX4,SLC40A1,Nrf2 and HO-1 were detected by Western blot in cells of 0,2.5,5 and 10 μmol/L E804 groups.Results Compared with the control group(0 μmol/L E804),2.5,5 and 10 μmol/L E804 significantly increased intracellular ROS,Fe2+and MDA levels,and decreased intracellular GSH content(P＜0.01).Meanwhile,the expression levels of SLC7A11,GPX4,SLC40A1,Nrf2 and HO-1 significantly decreased(P＜0.01),and the expression level of Transferrin increased(P＜0.05).Compared with the 10 μmol/L E804 group alone,the apoptosis inhibitor(Z-VAD)group and the ferroptosis inhibitor(DFO,Fer-1 and Lip-1)group could significantly reverse the inhibition of proliferation and migration of A549 cells by 10 μmol/L E804(P＜0.01).Conclution E804 can induce ferrop-tosis and inhibit the proliferation and migration of A549 cells,which may be related to the inhibition of Nrf2-HO-1/GPX4 pathway.

OBJECTIVES: To investigate the role of Holliday cross-recognition protein (HJURP) in tumorigenesis, progression, and immunotherapy responses. METHODS: Bioinformatics approaches were used to analyze the expression level of HJURP in various cancers and its association with prognosis, clinical stage, and immune cell infiltration using TCGA, GTEx, SangerBox and TIMER 2.0 databases. LinkedOmics database was employed to investigate HJURP-related genes and their potential functions in kidney renal clear cell carcinoma (KIRC). The expression of HJURP in KIRC samples was examined with immunohistochemistry, Western blotting and qRT-PCR, and the effect of HJURP silencing on cell proliferation and migration was tested in cultured KIRC cells. RESULTS: HJURP was highly expressed in 26 cancers with negative correlations with the patients' survival outcomes in 5 cancers including KIRC (P<0.05). HJURP expression levels was strongly correlated with clinical stages and immune cell infiltration in the tumors. In KIRC, HJURP expression was significantly elevated (P<0.0001) and showed a positive correlation with TNM stage (P<0.05), overall stage (P<0.01) and immune cell infiltration. Gene Ontology (GO) functional analysis showed that HJURP is predominantly enriched in biological processes such as biological regulation and metabolic processes. Concerning cellular components, HJURP is primarily localized to the cell membrane and nucleus. In terms of molecular functions, it is chiefly enriched in activities related to protein binding and ion binding. HJURP was highly expressed in both clinical KIRC tissues and KIRC cell lines (P<0.001). In cultured KIRC cells, silencing of HJURP significantly inhibited cell proliferation and migration abilities. CONCLUSIONS HJURP may serves as an indicator of prognosis and immunotherapy response of KIRC, and its high expression enhances malignant behaviors of KIRC cells.
Humans ; Prognosis ; Kidney Neoplasms/pathology* ; Biomarkers, Tumor/metabolism* ; Carcinoma, Renal Cell/pathology* ; DNA-Binding Proteins/genetics* ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

Microglial surveillance plays an essential role in clearing misfolded proteins such as amyloid-beta, tau, and α-synuclein aggregates in neurodegenerative diseases. However, due to the complex structure and ambiguous pathogenic species of the misfolded proteins, a universal approach to remove the misfolded proteins remains unavailable. Here, we found that a polyphenol, α-mangostin, reprogrammed metabolism in the disease-associated microglia through shifting glycolysis to oxidative phosphorylation, which holistically rejuvenated microglial surveillance capacity to enhance microglial phagocytosis and autophagy-mediated degradation of multiple misfolded proteins. Nanoformulation of α-mangostin efficiently delivered α-mangostin to microglia, relieved the reactive status and rejuvenated the misfolded-proteins clearance capacity of microglia, which thus impressively relieved the neuropathological changes in both Alzheimer's disease and Parkinson's disease model mice. These findings provide direct evidences for the concept of rejuvenating microglial surveillance of multiple misfolded proteins through metabolic reprogramming, and demonstrate nanoformulated α-mangostin as a potential and universal therapy against neurodegenerative diseases.

ObjectiveTo investigate the dietary preference and nutritional knowledge needs of the elderly people who dined at meal service sites. MethodsUsing the form of stratified and convenience sampling method with self-designed questionnaire was used， in November 2021， to select 700 elderly people who dine at meal service sites in 7 jurisdictions in Shanghai were selected， and a self-designed questionnaire was used to investigate the basic information. Results91.64% of the elderly surveyed would eat at relatively fixed meal service sites， and the total Dietary Diversity Score （DDS9） was 3.56±1.46. 41.45% of the elderly with diseases preferred unhealthy cooking methods. Only 8.03% of the surveyed seniors said they were unwilling to accept targeted and personalized nutrition tips and reminders. Multivariate logistic regression analysis showed that the probability reaching the “understanding” level of “Food Guide Pagoda for Chinese Residents” and “Four Principles Recommended by the Core Dietary Guidelines for the Elderly” was different in the elderly with different education levels. The willingness of the elderly to expect to receive different nutrition tips and reminders was related to whether they cared about the corresponding contents. There was a statistically significant difference （P<0.05） among the elderly who were concerned about different health problems in terms of the willingness to receive different nutritional tips. There were significant differences in the proportion of elderly people with different health status for intervention （χ²=5.402， P<0.05）. ConclusionThe elderly who dine at meal service sites are highly dependent on the sites， have a low level of dietary diversification， and do not have a high degree of understanding of nutrition-related knowledge， and have a high demand for targeted nutritional interventions. Nutritional interventions for the sick elderly should be piloted through multiple channels.

Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.

Objective:To investigate the predictive value of the National Institutes of Health Stroke Scale (NIHSS) score at 24 h after endovascular treatment on the outcomes in patients with acute basilar artery occlusion (ABAO).Methods:Consecutive patients with ABAO received endovascular treatment at the Department of Neurology, Shengli Oilfield Central Hospital from January 2019 to December 2020 were retrospectively included. According to the modified Rankin Scale scores at 90 days after onset, the patients were divided into a good outcome group (0-3) and a poor outcome group (4-6), as well as a survival group and a death group. The demographic and clinical data between the groups were compared respectively. Multivariate logistic regression analysis was use to identify independent influencing factors for clinical outcomes and mortality. The predictive value of postprocedural 24 h NIHSS score on the outcomes was evaluated using the receiver operating characteristic (ROC) curves. Results:A total of 35 patients with ABAO were included. Their age was 62 years (interquartile range, 56-66 years), and 28 patients were males (80%); 19 (54.3%) had a good outcome, 16 (45.7%) had a poor outcome, and 7 (20.0%) died. Univariate analysis showed that there were statistically significant differences in hypertension, low-density lipoprotein cholesterol, fasting blood glucose, collateral circulation grading, vascular recanalization, and postprocedural 24 h NIHSS scores between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that the postprocedural 24 h NIHSS score was independently correlated with the poor outcome (odds ratio 1.131, 95% confidence interval 1.017-1.258; P=0.023). Multivariate analysis did not find the independent influencing factors for death. ROC curve analysis showed that the area under the curve of the postprocedural 24 h NIHSS score for predicting poor outcome was 0.814 (95% confidence interval 0.668-0.960; P=0.011). The optimal cutoff value was 19 points, and the corresponding sensitivity and specificity were 85.7% and 71.4% respectively. Conclusions:In patients with ABAO receiving endovascular treatment, the postprocedural 24 h NIHSS score has good predictive value for poor outcomes at 90 d after procedure.