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 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.

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.

AIM:This study aims to investigate the mechanism by which Zhilianwan(ZLW)decoction affects dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)in mice,focusing on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.METHODS:Fifty male C57BL/6J mice were allowed ad libitum access to food for 1 week before the experiment.They were randomly divided into 5 groups:control group,model group,mesalazine(ME)group,high-dose ZLW(ZLW-H)group,and low-dose ZLW(ZLW-L)group,with ten mice in each group.All mice,except those in control group that received distilled water,were given a 3%DSS solution for seven consecutive days,starting the intervention on the first day of modeling.Each group re-ceived their respective treatments via gavage at volume of 20 μL/g for ten consecutive days.The mice in control and model groups received an equivalent volume of distilled water,while those in ME,ZLW-H,and ZLW-L groups were adminis-tered 0.2 g·kg-1·d-1 of ME,10.92 g·kg-1·d-1 of ZLW,and 2.73 g·kg-1·d-1 of ZLW,respectively.Body weight and dis-ease activity index(DAI)scores were recorded daily.At the end of the intervention,colon length was measured,and he-matoxylin-eosin staining was performed to assess pathological changes in colon tissues.ELISA and qPCR were employed to measure serum levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)and IL-10,as well as the mRNA expression levels in colon tissues.Western blot analysis was conducted to evaluate the protein levels of TLR4,MyD88,cytosolic NF-κB and nuclear NF-κB in colon tissues.RESULTS:Compared with model group,the mice treated with ZLW exhibited an increase in body weight and colon length(P＜0.05),along with a decrease in DAI and colon histo-pathological scores(P＜0.05).Serum levels of IL-1β,IL-6,and TNF-α were significantly lower(P＜0.05),while IL-10 levels increased(P＜0.05).The mRNA expression levels of these inflammatory factors in colonic tissues reflected similar trends.Additionally,levels of TLR4,MyD88,and nuclear NF-κB proteins were reduced(P＜0.05),whereas cytosolic NF-κB protein expression was elevated(P＜0.05)in the colon tissues of treated mice.CONCLUSION:ZLW exerts at-tenuating effects on DSS-induced intestinal injury in UC mice,though modulation of the TLR4/MyD88/NF-κB signaling pathway.

Objective·To investigate the role of HEN methyltransferase 1(HENMT1)in the proliferation and migration of gastric cancer(GC)and its potential molecular mechanisms.Methods·The expression of HENMT1 in GC was examined using bioinformatics databases,Western blotting and quantitative real-time PCR(qPCR).Kaplan-Meier Plotter and BEST online tools were used to analyze the correlations between HENMT1 expression and overall survival,perineural invasion,subtypes,tumor location and Lauren classification in clinical GC patients.GC cells were cultured in vitro and treated with small interfering RNA(siRNA)targeting HENMT1 and HENMT1 overexpression vectors,in combination with a PI3K activator(740 Y-P)or PI3K inhibitor(3-MA).The roles of HENMT1 in GC cell proliferation and migration were assessed using cell counting kit-8(CCK-8)assay,colony formation assay,wound healing assay and Transwell migration assay.Results·HENMT1 was significantly upregulated in GC and positively associated with perineural invasion.Its expression was closely related to GC subtypes,being most pronounced in the proliferative subtype,and was higher in intestinal-type GC according to the Lauren classification.However,HENMT1 expression showed no significant correlation with overall survival or tumor location(including gastric body,cardia,antrum and whole stomach).Functional experiments demonstrated that silencing HENMT1 inhibited GC cell proliferation and migration,whereas overexpression of HENMT1 enhanced these capabilities.Mechanistically,silencing HENMT1 reduced the levels of phosphorylated PI3K,AKT and mTOR,as well as their total protein expression.Conversely,HENMT1 overexpression upregulated these proteins.Moreover,siHENMT1 combined with the PI3K activator 740 Y-P effectively reversed the proliferation and migration effects induced by 740 Y-P,while overexpressed HENMT1 combined with the PI3K inhibitor 3-MA reversed the suppressive effects of 3-MA on GC cell proliferation and migration.Conclusion·HENMT1 is highly expressed in GC and positively regulates the proliferation and migration of gastric cancer cells by activating the PI3K-AKT-mTOR signaling pathway.

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 investigate the role of HEN methyltransferase 1(HENMT1)in the proliferation and migration of gastric cancer(GC)and its potential molecular mechanisms.Methods·The expression of HENMT1 in GC was examined using bioinformatics databases,Western blotting and quantitative real-time PCR(qPCR).Kaplan-Meier Plotter and BEST online tools were used to analyze the correlations between HENMT1 expression and overall survival,perineural invasion,subtypes,tumor location and Lauren classification in clinical GC patients.GC cells were cultured in vitro and treated with small interfering RNA(siRNA)targeting HENMT1 and HENMT1 overexpression vectors,in combination with a PI3K activator(740 Y-P)or PI3K inhibitor(3-MA).The roles of HENMT1 in GC cell proliferation and migration were assessed using cell counting kit-8(CCK-8)assay,colony formation assay,wound healing assay and Transwell migration assay.Results·HENMT1 was significantly upregulated in GC and positively associated with perineural invasion.Its expression was closely related to GC subtypes,being most pronounced in the proliferative subtype,and was higher in intestinal-type GC according to the Lauren classification.However,HENMT1 expression showed no significant correlation with overall survival or tumor location(including gastric body,cardia,antrum and whole stomach).Functional experiments demonstrated that silencing HENMT1 inhibited GC cell proliferation and migration,whereas overexpression of HENMT1 enhanced these capabilities.Mechanistically,silencing HENMT1 reduced the levels of phosphorylated PI3K,AKT and mTOR,as well as their total protein expression.Conversely,HENMT1 overexpression upregulated these proteins.Moreover,siHENMT1 combined with the PI3K activator 740 Y-P effectively reversed the proliferation and migration effects induced by 740 Y-P,while overexpressed HENMT1 combined with the PI3K inhibitor 3-MA reversed the suppressive effects of 3-MA on GC cell proliferation and migration.Conclusion·HENMT1 is highly expressed in GC and positively regulates the proliferation and migration of gastric cancer cells by activating the PI3K-AKT-mTOR signaling pathway.

AIM:This study aims to investigate the mechanism by which Zhilianwan(ZLW)decoction affects dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)in mice,focusing on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.METHODS:Fifty male C57BL/6J mice were allowed ad libitum access to food for 1 week before the experiment.They were randomly divided into 5 groups:control group,model group,mesalazine(ME)group,high-dose ZLW(ZLW-H)group,and low-dose ZLW(ZLW-L)group,with ten mice in each group.All mice,except those in control group that received distilled water,were given a 3%DSS solution for seven consecutive days,starting the intervention on the first day of modeling.Each group re-ceived their respective treatments via gavage at volume of 20 μL/g for ten consecutive days.The mice in control and model groups received an equivalent volume of distilled water,while those in ME,ZLW-H,and ZLW-L groups were adminis-tered 0.2 g·kg-1·d-1 of ME,10.92 g·kg-1·d-1 of ZLW,and 2.73 g·kg-1·d-1 of ZLW,respectively.Body weight and dis-ease activity index(DAI)scores were recorded daily.At the end of the intervention,colon length was measured,and he-matoxylin-eosin staining was performed to assess pathological changes in colon tissues.ELISA and qPCR were employed to measure serum levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)and IL-10,as well as the mRNA expression levels in colon tissues.Western blot analysis was conducted to evaluate the protein levels of TLR4,MyD88,cytosolic NF-κB and nuclear NF-κB in colon tissues.RESULTS:Compared with model group,the mice treated with ZLW exhibited an increase in body weight and colon length(P＜0.05),along with a decrease in DAI and colon histo-pathological scores(P＜0.05).Serum levels of IL-1β,IL-6,and TNF-α were significantly lower(P＜0.05),while IL-10 levels increased(P＜0.05).The mRNA expression levels of these inflammatory factors in colonic tissues reflected similar trends.Additionally,levels of TLR4,MyD88,and nuclear NF-κB proteins were reduced(P＜0.05),whereas cytosolic NF-κB protein expression was elevated(P＜0.05)in the colon tissues of treated mice.CONCLUSION:ZLW exerts at-tenuating effects on DSS-induced intestinal injury in UC mice,though modulation of the TLR4/MyD88/NF-κB signaling 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

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.