Objective: To investigate the changing trends for associations of anxiety and depressive symptoms with smartphone addiction among middle school students, so as to provide a scientific basis for preventing smartphone addiction in middle school students. Methods: From 2022 to 2023, a method of combining convenient sampling with cluster sampling was used to select 8 923 middle school students from 27 junior high schools and 3 senior high schools in a district of Shenzhen City between September 2022 (baseline, T1) and September 2023 (follow up, T2). The Smartphone Addiction Scale-Short Version (SAS-SV), Patients Health Questionnaire-9 Item (PHQ-9), and Generalized Anxiety Disorder 7-item Scale (GAD-7) were administered to assess smartphone addiction, anxiety and depressive symptoms. Mixed effects models were used to analyze the association of anxiety and depressive symptoms with smartphone addiction among middle school students. Results: From September 2022 to September 2023, the reported prevalence of smartphone addiction increased from 24.22% to 25.25% ( χ 2=45.71); and smartphone addiction scores [ 24.00 (16.00, 32.00),25.00(16.00, 33.00)], anxiety symptom scores [2.00(0.00, 7.00),3.00(0.00, 7.00)] and depressive symptom scores[3.00(0.00, 8.00),5.00(0.00, 9.00)] all significantly increased ( Z =-17.43, -42.38, -41.57) (all P <0.05). There were statistically significant difference in the distribution of anxiety and depression symptom levels among middle school students in 2022 and 2023 ( χ 2=85.15, 106.85, both P <0.05). After adjusting for covariates such as age, gender and family background, mixed effects models revealed dose response associations of anxiety and depressive symptoms with smartphone addiction among middle school students:mild anxiety symptom( OR =3.22), moderate to severe anxiety symptom ( OR =5.36), mild depressive symptom ( OR =3.32) and moderate to severe depressive symptom ( OR =6.13) were significantly associated with higher risks of smartphone addiction (all P <0.05). Interaction effect analysis found that co existing anxiety and depressive symptoms synergistically increased addiction risk by 5.60 times ( OR =5.60) compared to the asymptomatic group, with 32% of the combined risk attributable to their interaction ( S=1.64, AP =0.32)(both P < 0.05 ). Conclusions Anxiety and depressive symptoms are significantly associated with smartphone addiction, exhibiting a synergistic effect. Attention should be paid to emotional issues and smartphone addiction among middle school students.

Objective:To investigate the effects of targeted silencing of muscle blind-like protein 1 (MBNL1) gene on the proliferation, apoptosis and migration abilities of leukemia cell line K562 and their possible mechanisms.Methods:Single gene analysis was used to search for differences in MBNL1 gene expression between leukemia samples (173 cases) and healthy control samples (70 cases) in The Cancer Genome Atlas (TCGA) database. The data were updated in 2018. The logarithmic growth phase leukemia cell line K562 was taken and divided into sh-MBNL1 group (transfected with shRNA sequence with targeted silencing of MBNL1 gene), sh-NC group (transfected with corresponding negative control shRNA sequence) and blank control group (not transfected with shRNA sequence). Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of MBNL1, transforming growth factor β 1 (TGF-β 1) and Smad7 mRNA in each group of cells; Western blotting was used to detect the relative expression levels of cell migration-related proteins, apoptosis-related proteins, TGF-β 1, and Smad7 proteins; CCK-8 method was used to detect cell proliferation ability; Transwell method was used to detect cell migration ability. Results:In TCGA database, the relative expression level of MBNL1 gene in leukemia samples was higher than that in healthy control samples ( P < 0.05). The relative expression levels of MBNL1 protein in the sh-MBNL1 group, sh-NC group and blank control group were 0.71±0.11, 1.00±0.11 and 1.03±0.10, respectively, and the difference was statistically significant ( F = 7.78, P < 0.05); the relative expression level of MBNL1 protein in the sh-MBNL1 group was lower than that in the sh-NC group and blank control group (both P < 0.05). The results of CCK-8 assay showed that the cell proliferation ability of sh-MBNL1 group at 72 and 96 hours after transfection was higher than that of sh-NC group and blank control group (both P < 0.05). The Transwell method detection results showed that the number of cell membrane penetration in the sh-MBNL1 group, sh-NC group and blank control group were 666±135, 1 072±157 and 1 006±51, respectively, and the difference was statistically significant ( F = 9.40, P = 0.014); the number of cell membrane penetration in the sh-MBNL1 group was less than that in the sh-NC group and blank control group (both P < 0.05). The relative expression level of E-cadherin protein in the sh-MBNL1 group was higher than that in the sh-NC group and blank control group (both P < 0.01); the relative expression levels of Vimentin, Bax, caspase-3, TGF-β 1, and Smad7 proteins in the sh-MBNL1 group were lower than those in the sh-NC group and blank control group (all P < 0.01). The qRT-PCR detection results showed that the relative expression levels of TGF-β 1 mRNA and Smad7 mRNA in the sh-MBNL1 group were lower than those in the sh-NC group and blank control group (both P < 0.05). Conclusions:Silencing of MBNL1 gene can promote the proliferation of leukemia cell line K562, weaken its migration ability, and affect cell apoptosis. The mechanism may be related to the regulatory effect of TGF-β-Smad signaling pathway.

Transforming growth factor-β (TGF-β) /Smad signaling pathway is an important signaling pathway in human cells, which can regulate life activities such as cell growth, proliferation and apoptosis, and its signal transduction consists of ligand and receptor binding, intracytoplasmic signaling pathway, as well as interactions within the cell nucleus. This signaling pathway is closely related to the pathogenesis, drug resistance, recurrence and prognosis of acute leukemia. Exploring the potential of direct inhibitors, related therapeutic targets and natural extracts of this signaling pathway for the treatment of leukemia can provide new ideas for the study of biomarkers and targeted gene therapy for leukemia.

Pyroptosis is a programmed cell death mediated by the Gasdermin family of proteins, which is different from apoptosis and necrosis and accompanied by the release of a large number of pro-inflammatory factors. Leukemia is a malignant proliferative disease derived from hematopoietic stem cells, the pathogenesis and progression of which are related to many factors. In recent years, with the in-depth study on the mechanism of pyroptosis, it has been found that pyroptosis is closely related to the occurrence and development of leukemia. Further study of the mechanism of pyroptosis in leukemia can provide a new idea for the diagnosis and treatment of leukemia.

Objective:To investigate the effect of luteolin(Lut)on the proliferation and ferroptosis of human adriamycin(ADR)resistant chronic myeloid leukemia(CML)K562/ADR cells and the underlying molecular mechanism. Methods:K562 and K562/ADR cells were treated with different concentrations of ADR.The sensitivity of K562 and K562/ADR cells to ADR was evaluated by CCK-8 assay;the effect of different concentrations of Lut on the proliferation of K562/ADR cells was assessed by CCK-8 assay,and the half inhibitory concentration(IC50)was calculated for subsequent experiments;the morphological changes of the cells were observed by an inverted microscope;CCK-8 assay was used to examine the sensitizing effect of Lut on ADR;FCM assay was used to study the effect of Lut on the apoptosis of K562/ADR cells;fluorescence probe DCFH-DA,Fe2+colorimetric assay and glutathione(GSH)kit were used to detect the content of reactive oxygen species(ROS),Fe2+and GSH in K562/ADR cells,respectively;Western blotting was used to examine the expression levels of glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)in K562/ADR cells;the effects of Lut on the proliferation of K562/ADR cells,ROS content,GSH content,Fe2+content and GPX4 expression were studied after treatment with ferroptosis inhibitor Ferrostatin-1(Fer-1). Results:Compared with control group(cells treated with 0 μmol/L Lut),Lut significantly inhibited the proliferation of K562/ADR cells(P＜0.001),improved the chemosensitivity of K562/ADR cells to ADR(P＜0.05),increased apoptosis rate(P＜0.001)and ROS level(P＜0.05)of K562/ADR cells,reduced the GSH level(P＜0.001),and increase Fe2+content(P＜0.01).Compared with control group(cells treated with 0 μmol/L Lut),the protein expressions of GPX4,Nrf2 and HO-1 decreased with the increase of Lut concentration(P＜0.05).Compared with the Lut treatment alone,the inhibitory effect on the proliferation of K562/ADR cells induced by Lut was partially restored by Fer-1 intervention(P＜0.05),and intracellular ROS level was decreased(P＜0.001),GSH level was increased(P＜0.001),Fe2+content was decreased(P＜0.001)and the expression of GPX4 was increased(P＜0.01)in K562/ADR cells. Conclusion:Lut can inhibit the proliferation of K562/ADR cells through ferroptosis pathway,improve the chemosensitivity to ADR,and the potential mechanism may be related to the Nrf2/HO-1 signaling pathway,which provides experimental basis for the treatment of leukemia by ferroptosis.

Objective To investigate the mechanism of luteolin's(Lut)reversal effect on multidrug resistance of chronic myeloid leukemia K562/ADR cells.Methods CCK-8 assay was used to detect drug resistance in K562 and K562/ADR cells 24 hours after treatment with different doses of adriamycin(ADR).CCK-8 assay was used to assess the cytotoxicity and sensitizing effect of Lut on ADR after K562/ADR cells were treated with Lut alone or in combination with ADR for 24 hours.K562/ADR cells in logarithmic growth phase were separated into three group:0μmol/L Lut,2μmol/L and 4μmol/L Lut groups.ADR accumulation in cells was measured using flow cytometry.Nuclear factor erythroid-2-related factor 2(Nrf2),multidrug resistance associated protein 1(MRP1),P-glycoprotein(P-gp)and glutathione-S-transferase-PI(GST-pi)mRNA and protein expressions were identified using RT-PCR and Western blot assay.Glutathione(GSH)kit was used to detect intracellular GSH content.Results Compared with K562 cells,K562/ADR cell line was significantly resistant to ADR,and the drug resistance was 53.69 times.K562/ADR cell proliferation was decreased to variable degrees by different doses of Lut when compared to the 0μmol/L Lut group(P＜0.05).The proliferation inhibition rates of K562/ADR cells treated with 2 and 4μmol/L Lut were less than 10%,indicating that the concentration of Lut was non-toxic.Compared with the 0 μmol/L Lut group,the 2 μmol/L Lut group and the 4 μmol/L Lut group showed significantly increased ADR growth inhibition rate on K562/ADR and increased accumulation of ADR in cells,improved the reversal resistance fold,and decreased GSH content in cells.MRP1,P-gp,GST-pi and Nrf2 mRNA and protein expression were reduced in cells(P＜0.05).The effect of 4 mol/L Lut was greater than that of 2 mol/L Lut.Conclusion Lut may decrease K562/ADR cell proliferation and reverse ADR medication resistance.The mechanism could be connected to the downregulation of Nrf2,MRP1,P-gp and GST-pi expression,which leads to an increase in ADR accumulation in K562/ADR cells.

Leukemia is a group of hematologic malignancie. Ferroptosis is a novel of cell death caused by iron-dependent lipid peroxidation accumulation, which participates in the occurrence and development of leukemia. Activation of different regulatory sites in the ferroptosis pathway can promote the death of leukemia cells. Therefore, it can provide a new direction for the treatment of leukemia by inducing ferroptosis of cells.

Objective: To investigate the drug resistance of kaempferol reversed adriamycin (ADM)-resistant K562/ADM cells in chronic myelogenous leukemia (CML) and its related mechanism. Methods: Methyl thiazolyl tetrazolium (MTT) method was used to detect the toxicity of ADM on K562 and K562/ADM cells for 24 h. The half inhibitory concentration (IC₅₀) of ADM and the drug resistance multiple for 24 h were calculated. MTT method was used to detect the toxicity of kaempferol on K562/ADM cells for 24 h. The 5% inhibitory concentration (IC₅) and 10% inhibitory concentration (IC₁₀) of kaempferol for 24 h were calculated to determine the concentration of kaempferol in the subsequent experiments. And the cells untreated by the kaempferol were selected as the control group. The cell inhibition after the treatment of ADM for 24 h of the blank control group and kaempferol intervention group was detected by using MTT method. And then the cell inhibition for 24 h and ADM IC₅₀ for 24 h in the above groups were calculated. The ratio of IC₅₀ in the blank control group and kaempferol group was the reversal drug resistance multiple of kaempferol. The fluorescence intensity of ADM in K562/ADM cells treated by kaempferol was detected by using flow cytometry. Western blotting was used to detect the expressions of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), phosphorylated p38 (p-p38), and total p38 (t-p38) protein in K562/ADM cells after the treatment of kaempferol, the specific inhibitor of p38-MAPK signaling pathway SB202190, and the combination of kaempferol and SB202190. Results: After the treatment of ADM for 24 h, the IC₅₀ value of K562 and K562/ADM cells was (0.9±0.6), (28.1 ±3.5) μg/ml, respectively. The drug resistance multiple of K562/ADM cells on the treatment of ADM for 24 h was 31.16 compared with the K562 cells. MTT method showed that kaempferol inhibited the proliferation of K562/ADM cells in a dose-dependent manner. According to the IC₅ and IC₁₀, 0.5 μmol/L and 1.0 μmol/L kaempferol were determined to do the subsequent experiments. After the combined interaction of kaempferol and ADM for 24 h, the ADM IC₅₀ of K562/ADM cells in the blank control group, 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was (33.7±5.7), (21.4±0.6), (15.9±1.8) μg/ml, respectively (F = 30.85, P < 0.05), and there was a statistical difference of pairwise comparison (both P < 0.05). The reversal drug resistance multiple of K562/ADM cells for 24 h in 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was 1.58 and 2.12, respectively. Flow cytometry results showed that the mean fluorescence intensity (MFI) of ADM in the blank control group, 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was 138.4±8.9, 154.3±2.2, 165.7±4.8, respectively, and the difference was statistically significant (F = 161.48, P < 0.05). Compared with the blank control group, after treatment of K562/ADM cells with 0.5 μmol/L and 1.0 μmol/L kaempferol for 24 h, the relative expressions of P-gp, MRP1 and p-p38 protein were decreased in K562/ADM cells (all P < 0.05), but there was no statistical difference in the expression of t-p38 protein (P > 0.05); SB202190 could reduce the relative expressions of P-gp, MRP1 and p-p38 protein (all P < 0.05); after the treatment of SB202190 combined with different concentration of kaempferol, the relative expressions of P-gp, MRP1 and p-p38 protein in K562/ADM cells did not decrease (P > 0.05). Conclusions Kaempferol can decrease the relative expressions of P-gp and MRP1 in K562/ADM cells by inhibiting p38-MAPK pathway, so as to increase the concentrations of ADM and to reverse the drug resistance of K562/ADM cells.

Objective To explore the effect of proliferation and apoptosis of Solanine on acute T lymphocyte leukemia (T-ALL) Jurkat cells and its mechanism.Methods After treated with different concentrations of Solanine,the proliferation of Jurkat cells was detected by CCK-8 assay,and the effect of Solanine on apoptosis of Jurkat cells were detected by flow cytometry.The expression of Bcl-2 and Bax in Jurkat cells were detected by Western blot,and the expression levels of Bcl-2 mRNA and Bax mRNA were detected by real-time fluorescence quantitative polymerase chain reaction.Results CCK-8 assay showed that Solanine significantly inhibited the proliferation of Jurkat cells in a dose-and time-dependent manner.The results of flow cytometry showed that the apoptosis rates of Jurkat cells treated with Solanine for 24 h were (2.40-± 0.98) ％,(28.43-± 4.86) ％,(41.56-± 1.87) ％,respectively,in a dose-dependent manner.Western blot showed that Solanine could increase the expression of Bax and decrease the expression of Bcl-2 in Jurkat cells,and they all were dose-dependent.Conclusion Solanine can significantly inhibit the proliferation and induce apoptosis of Jurkat cells.The mechanism is related to the up-regulation of Bax expression and down-regulation of Bcl-2 expression.

Objective To evaluate the accuracy of MSCT in the pre-operative N-staging and diagnosis of metastatic lymph nodes in each group of gastric cancer.Methods Pathological and CT data of 91 patients with gastric cancer proved by surgery and pathology were analyzed retrospectively.Three-phase dynamic enhancements were performed before surgery in a unified way of hypotonic oral water,N-stage and grouping of lymph nodes of the preoperative CT imaging were evaluated by using the established diagnostic criteria and then compared with the results of surgery and pathology,the accuracy of staging and grouping was analyzed by using Kappa test.Results The accuracy of MSCT diagnosing the N-staging as a whole was 86.3%.The accuracy for N0,N1,N2 and N3 was 83.5%,89.0%,83.5% and 89.0%,respectively.The sensitivity was 86.5%,83.3%,50% and 47.4%,respectively.The specificity was 79.5%,89.4%,89.6% and 100.0%, respectively.The sensitivity for N0 was statistically different from that for N2, N0 and N3(P≤0.007).The detected accuracy for the group of left side of the cardium (No.2), periphery of the splenic hilum (No.10), posterior of the pancreastic head (No.13) were higher than other groups on MSCT with the accuracyof 100%.The sensitivity for the group of No.2,periphery of the coeliac trunk(No.9),No.10,and No.13 was 100%.The specificity for the group of No.2,No.10,and No.13 was 98.9%.Conclusion Relatively high accuracy in the preoperative N-staging and diagnosis of metastatic lymph nodes in each group of gastric cancer can be obtained by MSCT, which provide reliable information for preoperative assessment and intraoperative lymph node dissections.