Myelodysplastic syndromes (MDS) are associated with a high risk of progression to leukemia, highlighting the critical need to explore novel therapeutic agents. Based on network pharmacology prediction and molecular docking, we hypothesized that luteolin (LUT) might induce cellular senescence and apoptosis in MDS cells by targeting the Akt signaling pathway. This study aims to verify its targets and molecular mechanisms through experimental studies.

By integrating multiple databases, such as Pharm Mapper and Gene Cards, common targets of LUT and MDS were screened. A protein-protein interaction (PPI) network was constructed using the STRING database and visualized with Cytoscape software to identify key targets. The DAVID database was employed for the functional enrichment analysis of Gene Ontology (GO) and the pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) for the common targets. Molecular docking between LUT and the key targets was performed using Auto Dock software, and the docking efficacy was evaluated by analyzing the binding energy. Subsequently, molecular dynamics simulations were carried out using Gromacs to observe the ligand-receptor interactions and conformational changes. MUTZ-1 and MOLM-13 cells were used as experimental models. After treating the cells with different concentrations of LUT for 24 h (MUTZ-1:0、15、30、60 μmol/L, MOLM-13: 0、4、8、16 μmol/L), cell viability was measured by CCK-8 assay, DNA synthesis capacity was assessed by Ed U assay, colony formation was examined by soft agar assay, morphological changes were observed by transmission electron microscopy (TEM), intracellular reactive oxygen species (ROS), mitochondrial ROS, mitochondrial membrane potential, and apoptosis rates were detected by flow cytometry, senescence was assessed by senescence-associated β-galactosidase (SA-β-Gal) staining, m RNA expression levels of senescence-associated secretory phenotype (SASP) factors were determined by RT-q PCR, and protein expression levels were examined by Western blotting.

A total of 568 LUT targets and 2348 MDS-related targets were screened, yielding 228 intersecting targets. The intersecting targets were imported into the DAVID database. GO functional enrichment analysis demonstrated that biological processes were enriched in the responses to oxidative stress and cellular senescence; cellular components were enriched in protein kinase complex and secretory granule lumen; and molecular functions were enriched in protein kinase activity and cytokine receptor binding. KEGG pathway enrichment analysis revealed predominant enrichment in key pathways including the Akt signaling pathway, cellular senescence, and apoptosis. The PPI network constructed using the STRING database and Cytoscape software identified the top 10 key targets, as IL6, NFKB1, STAT3, TNF, JUN, BCL2, ALB, CASP3, Akt1 and IL1B. Molecular docking results indicated that the binding energy between LUT and Akt1 was -6.25 kcal/mol, suggesting a strong interaction. The molecular dynamics simulation results demonstrated stable binding between LUT and the Akt1 target. In vitro experimental results showed that LUT inhibited the proliferation of both MUTZ-1 and MOLM-13 cells in a dose-dependent manner. The half-maximal inhibitory concentration (IC_{5 0}) of LUT acting on MUTZ-1 cells for 24 h was 29.960 μmol/L, and that on MOLM-13 cells for 24 h was 8.654 μmol/L. Treatment of MUTZ-1 and MOLM-13 cells with various concentrations of LUT for 24 h significantly inhibited DNA synthesis and colony formation (P<0.01). The treatment of LUT at different concentrations for 24 h resulted in significant increases in intracellular ROS and mitochondrial ROS levels in a dose-dependent manner (P<0.05), decrease in mitochondrial membrane potential in a dose-dependent manner (P<0.05), and elevation of apoptosis rates in a dose-dependent manner (P<0.05). TEM revealed obvious apoptotic morphological features in MUTZ-1 and MOLM-13 cells after 24 h of LUT treatment. The protein levels of Cleaved PARP and Cleaved caspase-3, and the pro-apoptotic protein BAX were significantly upregulated, and that of the anti-apoptotic protein BCL-2 was significantly downregulated. SA-β-Gal staining showed that the proportion of senescent cells was significantly increased in MUTZ-1 and MOLM-13 cells after LUT treatment for 24 h, along with enhanced expression of related SASP factors (P<0.001), and significant upregulation of senescence-associated proteins p53, p21 and p16. After LUT treatment for 24 h, the protein expression of p-Akt (Ser473) and p-MDM2 (Ser166) was significantly downregulated, while the total Akt protein expression showed no significant change.

LUT may induce mitochondria-dependent apoptosis and senescence in MDS cells by targeting the Akt signaling pathway, thereby inhibiting the proliferation of MDS cells.

To explore the changes in serum energy metabolites in patients with peripheral T-cell lymphoma, and investigate serum biomarkers for monitoring peripheral T-cell lymphoma from the perspective of energy metabolism.

Multiple/selected reaction monitoring (MRM/SRM) was used to detect the energy-related metabolites in the sera of 16 patients with newly diagnosed peripheral T-cell lymphoma admitted in the Hematology Medical Center of the Second Affiliated Hospital of Army Medical University from November 2020 to December 2021, as well as 10 recruited healthy volunteers.The corresponding clinical data including medical history, laboratory results and image data were collected and retrospectively analyzed.

Significant differences were seen in the contents and expression profiles of serum energy metabolism-related products between the patients and the healthy volunteers. The patients had significantly reduced serum contents of cyclic AMP, succinate, citrate and cis-aconitate (P < 0.05), and elevated D-glucose 6-phosphate content (P < 0.05).The serum contents of citrate and succinate were negatively correlated with the risk stratification (low-, moderate-and high-risk) and clinical stage of the disease (P < 0.05).Meanwhile, there was a negative correlation between the contents of L-malic acid and citrate and the mid-term efficacy evaluation results, such as complete/partial response (CR/PR) or stable disease (SD)(P < 0.05).For patients with extranodal NK/T cell lymphoma (n=10), there were also significant reductions in the contents of cyclic AMP, succinate, citrate, isocitrate and cis-aconitate in the sera of patients compared with healthy volunteers (P < 0.05), and the contents of citrate and succinate were negatively correlated with the clinical stage (P < 0.05) and were rather correlated with mid-term efficacy evaluation results (CR/PR or SD)(P < 0.05).For patients with angioimmunoblastic T-cell lymphoma (n=6), the serum contents of cyclic AMP, citrate and succinate were significantly lower, while the content of D-glucose 6-phosphate was higher when compared with the healthy volunteers (P < 0.05), and the content of succinate was negatively correlated with both clinical stage and risk grade of the patients (P < 0.05).

There are 5 serum differential metabolites identified between patients with peripheral T-cell lymphoma and healthy controls, and succinate and citrate are expected to be serum biomarkers of peripheral T-cell lymphoma.

To observe the efficacy and safety of flumatinib conversion in chronic myelogenous leukemia-chronicphase (CML-CP) patients with suboptimal TKI response or intolerance.

Patients who did not have the best response or intolerance to first-line imatinib, dasatinib, and nilotinib and switched to flumatinib (600 mg/d) from February 2020 to August 2022 were collected from 5 hospitals from Chongqing and affiliated hospitals of North Sichuan Medical College.The efficacy and safety of flumatinib were observed.The optimal response rate, major molecular response (MMR), cumulative complete cytogenetic response (CCyR) rate, cumulative MMR rate, cumulative deep molecular response (DMR), progression-free survival (PFS), event-free survival (EFS) and adverse reactions in 3, 6 and 12 months after treatment were observed and analyzed.

A total of 100 patients with CML-CP were enrolled, with a median follow-up of 18(3~36) months.The optimal response rate was 92.6%(88/95), 94.4%(85/90) and 92.9%(79/85) respectively, at 3, 6 and 12 months after treatment.Till August 20, 2023, the cumulative CCyR and MMR rate was 98.0%(98/100) and 81.9%(77/94), respectively, the median time to reach CCyR and MMR was 3 months, and cumulative DMR rate was 51.0%(51/100).PFS rate was 100.0%(100/100) and 1-year EFS rate was 85.6%(75/90).The most common non-hematologic adverse reactions of flumatinib were diarrhea and abdominal pain (7.0%), followed by renal dysfunction (6.0%) and musculoskeletal pain (2.0%).The main hematologic adverse reactions were thrombocytopenia (12.0%), anemia (6.0%) and leukopenia (2.0%).

Flumatinib has better MMR and DMR and is well tolerated in CML-CP patients with TKI resistance or intolerance.

To preliminarily investigate the anti-tumor effects of phytosphingosine (PHS) and the involvement of inducing apoptosis of leukemia cells.

Cellular model of leukemia was established in leukemia cell lines K562 and SUP-B15.CCK-8 assay and EdU assay were used to measure the viability and DNA synthesis of K562 and SUP-B15 cells. RNA-seq was carried out to verify the differentially expressed genes (DEGs) after PHS treatment.Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were applied to analyze the involved functions and signaling pathways. Comparative Toxicogenomics Database (CTD) and Discovery Studio software were employed to predict the underlying targets of PHS and molecular docking.Cell apoptosis was detected by flow cytometry, mitochondrial membrane potential was evaluated by JC-1 probe, and protein expression of key molecules was validated by Western blotting.

PHS inhibited the proliferation of K562 and SUP-B15 cells in a time-and dose-dependent manner.The half-maximal inhibitory concentration (IC₅₀) of K562 cells was 17.67 and 12.52 μmol/L for 24 and 48 h, respectively, and the IC₅₀ value of SUP-B15 cells was 17.58 and 14.86 μmol/L for 24 and 48 h, respectively.PHS treatment at a dose of 20 μmol/L for 48 h resulted in significant inhibition of DNA synthesis.GO enrichment analysis of the K562 cells showed that PHS might be involved in positive regulation of apoptotic process, plasma membrane and its integral components, and protein kinase binding and activity.Reverse predictive analysis showed that BCL-2 protein was the most likely target of PHS.PHS significantly increased the apoptotic rate of leukemia cells (P < 0.05) in a dose-dependent manner, reduced the mitochondrial membrane potential, and down-regulated BCL-2 level (P < 0.05) and up-regulated the levels of Cleaved caspase-3 and Cleaved caspase-9(P < 0.05).

PHS may inhibit the proliferation of leukemia cells by inducing mitochondria-dependent apoptosis, possibly through PHS and BCL-2 interaction.