Xiaoaiping (XAP) Injection demonstrates the anti-prostate cancer (PCa) effects, yet the underlying mechanism remains unclear. This study aims to investigate the impact of XAP on PCa and elucidate its mechanism of action. PCa cell proliferation was evaluated using a cell counting kit-8 (CCK-8) assay. Cell apoptosis was assessed through Hoechst staining and Western blotting assays. Proteomics technology was employed to identify key molecules and significant signaling pathways modulated by XAP in PCa cells. To further validate potential key genes and important pathways, a series of assays were conducted, including acridine orange (AO) staining, transmission electron microscopy, and immunofluorescence assays. The molecular mechanism of XAP against PCa in vivo was examined using a PC3 xenograft mouse model. Results demonstrated that XAP significantly inhibited cell proliferation in multiple PCa cell lines. In C4-2 and prostate cancer cell line-3 (PC3) cells, XAP induced cellular apoptosis, evidenced by reduced B-cell lymphoma 2 (Bcl-2) levels and elevated Bcl-2-associated X (Bax) levels. Proteomic, immunofluorescence, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) investigations revealed a strong correlation between forkhead box O3a (FoxO3a) autophagic degradation and the anti-PCa action of XAP. XAP hindered autophagy by reducing the expression levels of autophagy-related protein 5 (Atg5)/autophagy-related protein 12 (Atg12) and enhancing FoxO3a expression and nuclear translocation. Furthermore, XAP exhibited potent anti-PCa action in PC3 xenograft mice and triggered FoxO3a nuclear translocation in tumor tissue. These findings suggest that XAP induces PCa apoptosis via inhibition of FoxO3a autophagic degradation, potentially offering a novel perspective on XAP injection as an effective anticancer therapy for PCa.
Male ; Humans ; Prostatic Neoplasms/physiopathology* ; Autophagy/drug effects* ; Animals ; Drugs, Chinese Herbal/pharmacology* ; Proteomics ; Mice ; Apoptosis/drug effects* ; Cell Line, Tumor ; Cell Proliferation/drug effects* ; Forkhead Box Protein O3/genetics* ; Xenograft Model Antitumor Assays ; Mice, Nude ; Mice, Inbred BALB C

Objective:To investigate the dynamic changes of cytokine levels in patients with sepsis and to identify potential biomarkers for evaluating the prognosis of the disease.Methods:A total of 195 patients with sepsis hospitalized at the Department of Infectious Diseases and the Department of Critical Care Medicine of the First Affiliated Hospital of Soochow University from August 2022 to October 2024 were recruited, and 70 healthy individuals undergoing physical examinations were recruited as the healthy control group. The levels of 11 cytokines, including interferon γ (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, tumor necrosis factor α (TNF-α) and C-reactive protein (CRP) were compared between the sepsis patients and the healthy controls. Spearman correlation analysis was used to assess the correlation between cytokine levels and sequential organ failure assessment (SOFA) scores in sepsis patients. Receiver operating characteristic curves were plotted and the area under the curve (AUC) was calculated to evaluate the diagnostic value of cytokines for sepsis. Delong test was used to compare AUC. Based on the 28-day survival outcomes, the sepsis patients were categorized into non-survival group and survival group. The levels of the 11 cytokines in patients on the 1st, 3rd, 7th, 14th, 21st and 28th days after confirmed sepsis were dynamically monitored, and their change characteristics were analyzed. Mann-Whitney U test was used for statistical comparison. Results:The age of the 195 patients with sepsis was 68.0 (55.0, 76.0) years old, including 124 males (63.6%), 64 died and 131 survived.The levels of IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, TNF-α, CRP in the sepsis group were all higher than those in the healthy control group ( Z=-2.99, -5.42, -4.95, -4.09, -5.05, -11.30, -8.66, -8.23, -5.64, -4.75, -2.12 and -10.75, respectively, all P<0.05). The differences were statistical significance. The levels of IL-2 ( r=0.149, P=0.037), IL-6 ( r=0.223, P=0.002), IL-8 ( r=0.159, P=0.026), and IL-10 ( r=0.188, P=0.009) in patients with sepsis were positively correlated with SOFA scores. The AUC of CRP in diagnosing sepsis was 0.989 with the sensitivity of 97.4% and the specificity of 100.0%. The AUC of IL-6 in diagnosing sepsis was 0.953, with the sensitivity of 93.3% and the specificity of 97.1%, and the AUC of IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12P70, IL-17A, and TNF-α were 0.620, 0.718, 0.699, 0.665, 0.703, 0.850, 0.836, 0.727, 0.691, and 0.574, respectively.The AUC of the 11 cytokines were all lower than that of CRP, and the differences were all statistically significant ( Z=2.34, 10.24, 8.03, 8.08, 10.64, 8.70, 5.91, 5.17, 8.91, 9.25 and 4.10, respectively, all P<0.05).During the dynamic monitoring, the IFN-γ and IL-1β levels in the non-survival group increased gradually. The IFN-γ levels on the 14th and 21st day in the non-survival group were higher than those in the survival group ( Z=0.53 and 0.08, respectively, both P<0.05), and IL-1β levels on the 14th, 21st, and 28th days were also higher than those in the survival group ( Z=0.03, 0.26 and 0.31, respectively, all P<0.05). IL-6 and IL-8 levels reached their peaks on the 14th day, which were significantly higher than those in the survival group ( Z=0.01 and 0.02, respectively, both P<0.05), and then decreased, and the differences were all statistically significant. Conclusions:The levels of IFN-γ and IL-1β in the non-survival sepsis patients show a gradually increasing trend. The dynamic changes of IL-6 have certain significance for the prediction of disease severity and prognosis evaluation in sepsis.

Objective:To investigate the dynamic changes of cytokine levels in patients with sepsis and to identify potential biomarkers for evaluating the prognosis of the disease.Methods:A total of 195 patients with sepsis hospitalized at the Department of Infectious Diseases and the Department of Critical Care Medicine of the First Affiliated Hospital of Soochow University from August 2022 to October 2024 were recruited, and 70 healthy individuals undergoing physical examinations were recruited as the healthy control group. The levels of 11 cytokines, including interferon γ (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, tumor necrosis factor α (TNF-α) and C-reactive protein (CRP) were compared between the sepsis patients and the healthy controls. Spearman correlation analysis was used to assess the correlation between cytokine levels and sequential organ failure assessment (SOFA) scores in sepsis patients. Receiver operating characteristic curves were plotted and the area under the curve (AUC) was calculated to evaluate the diagnostic value of cytokines for sepsis. Delong test was used to compare AUC. Based on the 28-day survival outcomes, the sepsis patients were categorized into non-survival group and survival group. The levels of the 11 cytokines in patients on the 1st, 3rd, 7th, 14th, 21st and 28th days after confirmed sepsis were dynamically monitored, and their change characteristics were analyzed. Mann-Whitney U test was used for statistical comparison. Results:The age of the 195 patients with sepsis was 68.0 (55.0, 76.0) years old, including 124 males (63.6%), 64 died and 131 survived.The levels of IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, TNF-α, CRP in the sepsis group were all higher than those in the healthy control group ( Z=-2.99, -5.42, -4.95, -4.09, -5.05, -11.30, -8.66, -8.23, -5.64, -4.75, -2.12 and -10.75, respectively, all P<0.05). The differences were statistical significance. The levels of IL-2 ( r=0.149, P=0.037), IL-6 ( r=0.223, P=0.002), IL-8 ( r=0.159, P=0.026), and IL-10 ( r=0.188, P=0.009) in patients with sepsis were positively correlated with SOFA scores. The AUC of CRP in diagnosing sepsis was 0.989 with the sensitivity of 97.4% and the specificity of 100.0%. The AUC of IL-6 in diagnosing sepsis was 0.953, with the sensitivity of 93.3% and the specificity of 97.1%, and the AUC of IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12P70, IL-17A, and TNF-α were 0.620, 0.718, 0.699, 0.665, 0.703, 0.850, 0.836, 0.727, 0.691, and 0.574, respectively.The AUC of the 11 cytokines were all lower than that of CRP, and the differences were all statistically significant ( Z=2.34, 10.24, 8.03, 8.08, 10.64, 8.70, 5.91, 5.17, 8.91, 9.25 and 4.10, respectively, all P<0.05).During the dynamic monitoring, the IFN-γ and IL-1β levels in the non-survival group increased gradually. The IFN-γ levels on the 14th and 21st day in the non-survival group were higher than those in the survival group ( Z=0.53 and 0.08, respectively, both P<0.05), and IL-1β levels on the 14th, 21st, and 28th days were also higher than those in the survival group ( Z=0.03, 0.26 and 0.31, respectively, all P<0.05). IL-6 and IL-8 levels reached their peaks on the 14th day, which were significantly higher than those in the survival group ( Z=0.01 and 0.02, respectively, both P<0.05), and then decreased, and the differences were all statistically significant. Conclusions:The levels of IFN-γ and IL-1β in the non-survival sepsis patients show a gradually increasing trend. The dynamic changes of IL-6 have certain significance for the prediction of disease severity and prognosis evaluation in sepsis.