Objective: To investigate the therapeutic effects of human umbilical cord mesenchymal stem cell derived exosomes (hucMSC-Exo) on acute myocardial infarction (AMI) and to elucidate the underlying mechanisms. Methods: Human umbilical cord mesenchymal stem cells (hucMSCs) were isolated and cultured. Exosomes were extracted from the culture media. AMI model was constructed by ligating the anterior descending branch of the coronary artery in mice. The mice were randomly divided into Sham, AMI, Exo and Fer-1 groups, and PBS, PBS, hucMSC-Exo and Ferrostatin-1 were injected into the marginal zone of the ischemic myocardium of each group, respectively. The levels of the antioxidants glutathione (GSH) content and malondialdehyde (MDA) content in mouse myocardial tissues measured by corresponding assay kits, and reactive oxygen species (ROS) levels detected by DHE probe were used to assess the severity of ferroptosis in mouse myocardial tissue; Cardiac ultrasound and Masson staining were used to evaluate the cardiac function and fibrosis level in mice; HL-1 cells were treated with DMSO, Erastin and hucMSC-Exo and subjected to RNA sequencing, and the potential mechanism of the therapeutic effect of hucMSC-Exo was evaluated by KEGG enrichment analysis and clustering heat map analysis. The viability of HL-1 cells after hucMSC-Exo or hucMSC-Exo+BI-6C9 co-treatment was detected by CCK-8 assay. BH3 interacting domain death agonist (Bid) expression levels in infarcted myocardial tissues of mice were detected by qRT-PCR. Results: Compared with the AMI group, the Exo and Fer-1 groups of mice showed a significant decrease in MDA and ROS levels in the infarcted region of the myocardium, an increase in GSH content, a reduction in myocardial fibrosis, and an improvement in cardiac function; among the mice in the Exo group, the treatment effect was more pronounced compared to the Fer-1 group on the 28th day after AMI. KEGG enrichment analysis showed that the reversed genes by hucMSC-Exo were mainly enriched in the p53 signaling pathway; clustering heatmap analysis of p53-related genes revealed that Bid gene expression was significantly higher in HL-1 cells in the Erastin group compared with the control group, and the level of Bid expression in HL-1 cells in the Exo group was significantly lower than that in the Erastin group. The therapeutic effect of hucMSC-Exo in HL-1 cells was not significantly increased after treatment with the Bid inhibitor BI-6C9. Bid mRNA level in the heart tissues of mice in the AMI group was substantially increased compared with that in the control group, and was significantly decreased in the Exo group compared with that in the AMI group. Conclusion: hucMSC-Exo inhibits Bid expression to mitigate ferroptosis after AMI, thereby alleviate myocardial injury.

Objective: To explore the effect of neural stem cell secretome (NSC-S) on ferroptosis human neuroblastoma SH-SY5Y cells and its underlying mechanism. Methods: Human neuroblastoma SH-SY5Y cells were used to establish neuronal ferroptosis model in vitro. And the cells were divided into three groups: control group, Erastin group and Erastin+NSC-S group; cells were treated with DMEM, DMEM containing 10 μmol/L Erastin, and NSC-S containing 10 μmol/L Erastin, respectively. The activities of lactate dehydrogenase (LDH), intracellular reactive oxygen species and malondialdehyde in the supernatants of cells in each group were detected by commercial kits, and the contents of free ferrous ions were detected by fluorescent probes. The expression level of glutathione peroxidase 4 (GPX4) protein in cells was determined by Western blotting. The components of NSC-S were detected by liquid chromatography tandem mass spectrometry (LC-MS/MS) and bioinformatics analysis and screening were conducted. In addition, SH-SY5Y cells were divided into four groups, which were control group, Erastin group, Erastin+NSC-S group, and Erastin+Parkinson′s disease protein 7 (PARK7) group. And cells were treated with DMEM, DMEM containing 10 μmol/L Erastin, NSC-S containing 10 μmol/L Erastin, and DMEM containing 200 μg/L PARK7+10 μmol/L Erastin, respectively. The viability of SH-SY5Y cells was detected by the CCK-8 assay, and the contents of reactive oxygen species, malondialdehyde, ferrous ions in the cells as well as the expression level of GPX4 protein were also detected. Results: Compared with the Erastin group, the LDH activity of SH-SY5Y cells supernatant in the Erastin+NSC-S group was significantly decreased (P<0.001), the contents of intracellular reactive oxygen species, malondialdehyde and ferrous ions were significantly reduced (P<0.001 or P<0.05), and the relative expression level of GPX4 protein was greatly increased (P<0.05). LC-MS/MS analysis revealed that NSC-S contained PARK7 protein. Compared with the Erastin group, the viability of SH-SY5Y cells in the Erastin+PARK7 group was significantly enhanced (P<0.001), the contents of reactive oxygen species, malondialdehyde and ferrous ions were significantly decreased (P<0.001), and the relative expression level of GPX4 protein was significantly increased (P<0.05). Conclusion: NSC-S could inhibit ferroptosis induced by Erastin in human neuroblastoma SH-SY5Y cells, which may be related to the mediation of PARK7 in NSC-S.

Objective: To explore the regulatory role and mechanism of high mobility group box 1 (HMGB1) in the radiosensitivity of triple-negative breast cancer (TNBC) cells. Methods: The expression level of HMGB1 in breast cancer subtypes and its relationship with the survival prognosis of breast cancer patients were analyzed by using The Cancer Genome Atlas (TCGA). Breast epithelial cells MCF-10A, non-TNBC cells MCF-7, SK-BR-3, and TNBC cells MDA-MB-231, MDA-MB-436, MDA-MB-468 were cultured in vitro. The mRNA and protein expression levels of HMGB1 were detected by qRT-PCR and Western blotting. Cell proliferation was assessed at different time points (0, 24, 48, 72 and 96 h) after 4 Gy X-ray irradiation, and the radiosensitivity of the cells was evaluated after different doses (0, 2, 4, 6, 8 Gy) of irradiation. HMGB1 expression was knocked down or overexpressed in MDA-MB-231 cells using plasmids or lentivirus, respectively, and cell proliferation, clone formation ability and apoptosis level after 4 Gy X-ray irradiation were detected by CCK-8 assay, colony formation assay, and flow cytometry, respectively. Changes in nuclear and intracellular HMGB1 in MDA-MB-231 cells after 0, 4, and 8 Gy irradiation were detected by immunofluorescence assay. MDA-MB-231 cells were divided into shControl, shHMGB1-2, Vector, and Flag-HMGB1 groups, irradiated with 4 Gy for 72 h, and changes in nuclear, HMGB1, and γ-H2AX were detected by immunofluorescence assay. Protein expression of p-AKT, AKT, p-GSK-3β, GSK-3β, Caspase 9, p-H2AX, and HMGB1 were detected by Western blotting in MDA-MB-231 cells after 0, 4, 8 Gy irradiated, and in shControl, shHMGB1-2, Vector, Flag-HMGB1 groups cells after 4 Gy irradiation for 72 h. The proliferation inhibition rate was assessed by CCK-8 assay in control, radiotherapy, AKT inhibitor, and AKT inhibitor + radiotherapy groups. Cell proliferation activity was examined by CCK-8 assay, and protein expression of p-AKT/AKT, Caspase 9, and p-H2AX were detected by Western blotting in Vector, Flag-HMGB1, Vector+AKT inhibitor, and Flag-HMGB1+AKT inhibitor groups after 4 Gy irradiation. Results: HMGB1 was highly expressed in TNBC, and its high expression was associated with poor prognosis. The mRNA and protein expression levels of HMGB1 was higher in MDA-MB-231/436/468 cells than in MCF-7 and SK-BR-3 cells, with the former having lower radiosensitivity. Knockdown of HMGB1 significantly increased the radiosensitivity of MDA-MB-231 cells, while overexpression of HMGB1 significantly decreased it. Knockdown of HMGB1 resulted in a significant decrease in p-AKT expression levels and a significant increase in p-GSK-3β and p-H2AX expression levels post 4 Gy irradiation. Overexpression of HMGB1 resulted in a significant increase in p-AKT expression levels and a significant decrease in p-GSK-3β and pH2AX expression levels post 4 Gy irradiation. The resistance-promoting effect of HMGB1 on radiotherapy was abolished by AKT inhibitor AKTi-1/2 treatment. Conclusion: HMGB1 is highly expressed in TNBC cells and regulates the radio-sensitivity of cells through the PI3K/AKT pathway. Thus, it may be implicated that HMGB1 may potentially serve as an important molecular target for radio-resistance in TNBC.

Objective: To investigate the therapeutic effect and mechanism of Clitoria ternatea flower (CF) on nonalcoholic fatty liver disease (NAFLD) in mice induced by a highfat diet (HFD). Methods: Twentyfour healthy ICR mice were randomly divided into four groups: normal control group (NC group), NAFLD model group (NAFLD group), highdose CF group (CFH group, 200 mg/kg), and lowdose CF group (CFL group, 100 mg/kg), with 6 mice in each group. The NC group was fed a normal diet, while the NAFLD, CFH, and CFL groups were fed a 60% highfat diet to induce NAFLD. Additionally, the CFH and CFL groups received corresponding doses of CF by gavage, and the NC and NAFLD groups received an equal volume of normal saline (06 mL) by gavage daily for 12 weeks. Serum levels of hepatic enzymes, blood lipids, and oxidative stress factors were detected, combined with untargeted metabolomics analysis. Fecal microbiota composition was analyzed via 16S rRNA gene sequencing, and the contents of three major shortchain fatty acids (acetate, propionate, and butyrate) were determined. Meanwhile, hepatic inflammationrelated indicators were examined by quantitative realtime polymerase chain reaction (qRTPCR). Results: Compared with the NAFLD group, the CFL and CFH groups showed significant reductions in serum levels of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (TC), and lowdensity lipoprotein cholesterol (LDLC) (all P<005). In the liver, the mRNA expressions of proinflammatory cytokines IL1β and TNFα were decreased, while those of antiinflammatory cytokines IL4 and IL10 were increased (all P<001). Additionally, serum superoxide dismutase (SOD) activity was markedly enhanced, whereas malondialdehyde (MDA) content was significantly decreased (all P<001). 16S rRNA sequencing results indicated that compared with the NAFLD group, the CF groups had increased abundances of multiple beneficial gut bacteria including Prevotella and Lactobacillus, along with elevated concentrations of acetate, propionate, and butyrate in feces. Serum untargeted metabolomics analysis revealed that compared with the NAFLD group, the CF groups had higher levels of various antiinflammatory and antioxidative stress metabolites, which were mainly involved in glycerophospholipid metabolism, pyrimidine metabolism, and other pathways. Conclusion: CF can alleviate liver damage in NAFLD mice by improving lipid levels and hepatic inflammation, regulating gut microbiota balance and serum metabolic pathways, showing potential therapeutic effects for NAFLD.

Objective: To investigate the effect of 3,3′-diindolylmethane (DIM) on paclitaxelresistant nonsmall cell lung cancer A549/T cells and explore its underlying mechanism. Methods: Western blotting was used to detect transferrin receptor (TFRC), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in human normal bronchial epithelial BEAS-2b cells, human NSCLC A549 cells, paclitaxel-resistant human NSCLC A549/T cells, and human large cell lung cancer H460 cells to select the optimal cell line. A549 and A549/T cells were treated with various concentrations of DIM (0, 20, 40, 80, 160 μmol/L), and cell proliferation was assessed by CCK-8 assay; and the expression of TFRC, SLC7A11 and GPX4 proteins in A549/T cells were determined by Western blotting to screen out the optimal DIM concentration. Additionally, A549/T cells were divided into the control group, which was cultured in highglucose medium containing 10% fetal bovine serum for 24 h; the Erastin group, treated with 10 μmol/L Erastin for 12 h alone; the DIM group, treated only with 80 μmol/L DIM for 24 h; the DIM+Erastin group, cells were pre-treated with 10 μmol/L Erastin for 12 h, then treated with 80 μmol/L DIM in the following 24 h. Cell viability was detected by CCK8 assay, and the expression of TFRC, SLC7A11 and GPX4 proteins was assessed by Western blotting. Results: Compared with the BEAS-2b group, TFRC and SLC7A11 expression was significantly increased in A549 cells (P<0.05), TFRC, SLC7A11 and GPX4 expression was significantly increased in A549/T cells (P<0.05). No significant differences in TFRC, SLC7A11 and GPX4 expression were observed in H460 cells (P>005). In A549/T cells, compared with the 0 μmol/L group, the expression of TFRC, SLC7A11 and GPX4 proteins showed significant alterations in the 80 μmol/L DIM group (all P<0.05). Compared with 0 μmol/L DIM group, in A549 cells, cell viability was significantly decreased in 20, 40, 80 and 160 μmol/L DIM groups (all P<0.05); in A549/T cells, cell proliferation was significantly decreased in 80 and 160 μmol/L DIM groups (all P<0.05). In A549/T cells, compared with the control group, DIM group, Erastin group and DIM+Erastin group showed a significant increase in TFRC expression (all P<0.05), while the expression of SLC7A11 and GPX4 proteins significantly decreased (all P<0.05), and cell proliferation significantly decreased (all P<0.05). Conclusion: DIM may inhibit the proliferation of paclitaxel-resistant non-small cell lung cancer A549/T cells by inducing ferroptosis, and ferroptosis inducers could be used to enhance the anti-cancer effect of DIM.

Objective: To investigate the impact of lipotoxic hepatocytederived exosome (LTH-ex) on ferroptosis and activation of hepatic stellate cell (HSC). Methods: Lipotoxic damage was induced in the human liver cell line LO2 using oleic acid and palmitic acid. The supernatants of the lipotoxic damaged liver cells were collected and LTH-ex was extracted through ultracentrifugation. The characterization of LTH-ex was identified using transmission electron microscopy and Nanosight nanoparticle analysis system, including assessing morphology, particle size and concentration, and LTH-ex surface markers through Western blotting. The human HSC cell line LX-2 was cocultured with LTH-ex or the ferroptosis activator Erastin for 48 h, and then all cells were divided into three groups: the PBS control group, the LTH-ex treatment group, and the co-treatment group consisting of LTH-ex and the ferroptosis inducer Erastin. Cell viability and mitochondrial membrane potential of LX-2 were evaluated using FDA and JC-1 staining. The iron ion concentration kit was used to detect the iron level in LX-2, the expression of mRNA and protein of death suppressor genes GPX4, xCT, and HSC activation gene α-SMA, were measured through qRT-PCR and Western blotting. Additionally, a mouse model of nonalcoholic fatty liver disease was established by feeding a high-fat diet, followed by injection of LTH-ex through the tail vein. Liver tissue structure was observed using HE staining, while GPX4 and α-SMA protein expression was assessed through immunohistochemistry staining. Sirius red staining was employed to detect collagen deposition in the liver tissue. Results: The LTH-ex particles were approximately 110 nm in size, displaying the typical lipid bilayer structure of exosomes and containing exosome marker proteins TSG101, Alix, and CD63. In comparison to the PBS control group, the LTH-ex treatment group demonstrated enhanced viability of LX-2 cells, an increased mitochondrial membrane potential, decreased iron levels, and upregulation of the ferroptosis-inhibiting genes GPX4 and xCT, as well as HSC activation gene α-SMA. Conversely, when compared to the LTH-ex treatment group, the co-treatment group of LTH-ex and the ferroptosis inducer Erastin exhibited reduced LX-2 cell viability and mitochondrial membrane potential, alongside downregulation of GPX4, xCT, and α-SMA. HE staining, immunohistochemistry, and Sirius red staining demonstrated that LTH-ex could exacerbate vacuolar degeneration of hepatocytes in the liver tissue of mice with nonalcoholic fatty liver disease. Additionally, LTH-ex promoted the expression of GPX4 and αSMA proteins and increased collagen deposition in the liver tissue. expression. Conclusion: LTH-ex increase the cell viability of hepatic stellate cells and the expression of ferroptosis inhibitory genes GPX4 and xCT, and it may be implicated that LTH-ex may promote hepatic stellate cell activation by inhibiting ferroptosis, and exacerbate collagen deposition in the liver tissue of non-alcoholic fatty liver disease mice.

Objective: To figure out the possible mechanisms of ferroptosis and explore potential ferroptosisrelated genes (FRGs) biomarkers and pharmacological compounds for atherosclerosis (AS). Methods: The AS transcriptome dataset GSE100927 was downloaded from the gene expression omnibus (GEO) database. FRGs were downloaded from FerrDb database. To screen the ferroptosisrelated hub genes in AS (FRG-hubs) by weighted gene coexpression network analysis, differential gene analysis, LASSO regression and random forest algorithm. Singlesample gene set enrichment (ssGSEA) analysis was used to evaluate the immunological landscape. Furthermore, transcription factors and miRNAs regulatory networks were constructed by the NetworkAnalyst database, and candidate drugs were searched from DSigDB database. The expression of FRGhubs was verified by AS model mice. Results: zinc finger E-Box binding homeobox 1 (ZEB1), mitogenactivated protein kinase kinase kinase 11 (MAP3K11), and cyclin dependent kinase inhibitor 2A (CDKN2A) were identified as FRG-hubs, and the nomogram model based on them demonstrated high reliability and effectiveness. In addition, FRG-hubs and immune cell infiltration showed a significant association. AS patients could be classified into 2 ferroptosis-related cluster with different immune cell infiltration. Predictions were made for 13 transcription factors, 8 miRNAs, and 10 drugs targeting FRGhubs. The results of qRT-PCR and Western blotting show that compared with the control group, the mRNA and protein expression levels of Map3k11 and Cdkn2a are significantly elevated in the AS model mice, while the expression level of the Zeb1 is significantly decreased. Conclusion: ZEB1, MAP3K11 and CDKN2A may be involved in the occurrence and development of AS by regulating immune-related pathways through ferroptosis.

Objective: To investigate the role and mechanism of celastrol in the treatment of relapsed/refractory diffuse large B-cell lymphoma (DLBCL) by inducing ferroptosis. Methods: The rituximab-resistant lymphoma cell line SU-DHL-2-R was established by the concentration gradient method. The CCK-8 assay was used to determine cell viability after treatment with different concentrations of celastrol. SU-DHL-2-R cells were divided into control group, celastrol group, ferroptosis inhibitor (Fer-1) group, and celastrol+Fer1 group. qRT-PCR and Western blotting were used to detect the expression levels of ferroptosis-related genes SLC7A11 and GPX4. GSH and Fe2+ levels were detected using commercial kits, and reactive oxygen species (ROS) levels were detected using the DCFH-DA probe method. The protein expressions of PI3K, p-PI3K, Akt and pAkt were detected by Western blotting. SU-DHL-2-R cells were injected subcutaneously into SCID mice to establish lymphoma tumor bearing mouse models. The tumor bearing mice were divided into control and celastrol treatment groups. The survival status of the mice was monitored and tumor volume was measured. The expressions of SLC7A11 and GPX4 in the lymphoma tissues of the mice were detected by immunohistochemistry. Results: The SU-DHL-2-R cell model was successfully established, and the drug resistance index was 12. Celastrol significantly inhibited the proliferation of SU-DHL-2 and SU-DHL-2-R cells. Compared with the control group, the mRNA and protein expressions of SLC7A11 and GPX4 were downregulated, the level of GSH was decreased, ROS accumulation was induced, Fe2+ levels were increased, and the protein expression levels of pPI3K and p-Akt were significantly downregulated in the celastrol group. However compared with the celastrol group, the above changes were attenuated in the celastrol+Fer-1 group. The tumor-bearing mouse model of lymphoma was established. Compared with the control group, the tumor volume of mice treated with celastrol was reduced, and the expression of SLC7A11 and GPX4 was significantly decreased. Conclusion: Celastrol may inhibit the progression of relapsed/refractory DLBCL by inducing ferroptosis through targeting the PI3K/Akt pathway.

Objective: To investigate the origin and distribution of IL-9 derived cells in patients with different types of thyroid cancer. Methods: Patients with papillary thyroid carcinoma (22 cases) and medullary thyroid carcinoma (8 cases) were recruited from March 2021 to December 2023, along with 30 healthy individuals undergoing physical examinations as controls. Blood samples were collected preoperatively and within 7 days postoperatively from the thyroid cancer patients and the controls. The expression of IL-9 in peripheral blood mononuclear cells (PBMCs) was measured using flow cytometry. The levels of cytokines IL-13, IL-4, IL-33, IL-25, IL-1β, IL-9, and IL-21 in serum were determined through enzymelinked immunosorbent assay (ELISA). The mRNA levels of PU-1, SMAD2, TGFβ, and STAT3 in blood cells were measured via quantitative realtime PCR (qRTPCR). During surgeries, thyroid cancer tissue and adjacent normal tissue samples were obtained. The mRNA expression levels of IL-25, IL-33, IL-1β, IL-13, RORα, and IL-9 in tissues were detected by qRT-PCR. Immunofluorescence staining was applied to analyze IL-9 expression in cancerous and adjacent normal tissues. Results: Flow cytometry showed that the distribution frequency of IL-9-producing T cells (Th9 cells) was higher in papillary thyroid carcinoma patients than those in medullary thyroid carcinoma patients, and higher in medullary thyroid carcinoma patients than those in the control group. After surgery, the distribution frequency of Th9 cells increased significantly in both papillary thyroid carcinoma and medullary thyroid carcinoma groups. However, the distribution of type 2 innate lymphoid cells (ILC2s) producing IL9 showed no significant difference among the groups. Preoperatively, papillary thyroid carcinoma and medullary thyroid carcinoma patients had significantly higher serum levels of IL-9, IL-33, IL-25, IL-1β, IL-4, and IL-21 than the control group. Postoperatively, IL-9 and IL-4 levels increased further, while IL-33 and IL-25 levels decreased significantly. qRT-PCR revealed that the mRNA expression of Th9related transcription factors PU-1, SMAD2, TGFβ, and STAT3 was lowest in the control group and increased significantly after surgery in thyroid cancer patients. In cancer tissue, qRT-PCR showed that IL-25 and IL-33 mRNA expression in medullary thyroid carcinoma and papillary thyroid carcinoma were significantly higher than those in adjacent tissues, and the medullary thyroid carcinoma was significantly higher than that in papillary thyroid carcinoma tissues. Contrastedly IL-9 and IL-1β mRNA expression were decreased. Immunofluorescence staining showed that IL-9 expression was lower in cancer tissue than in adjacent normal tissue and was mainly located in CD4⁺ T cells. Conclusion: IL-9 derived from Th9 plays an antitumor role in patients with thyroid cancer. The presence and distribution of Th9 cells correlate with the malignancy of thyroid cancer. IL-9 and Th9 cell may be considered as promising new targets for the diagnosis and therapeutic monitoring of thyroid cancer.

Objective: To investigate the therapeutic effects and underlying mechanisms of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles (HucMSC-sEVs) on renal injury in diabetic kidney disease (DKD) rats. Methods: HucMSCs were isolated, cultured, and used to extract HucMSC-sEVs from their culture supernatant. A DKD rat model was induced by a high-fat diet combined with STZ injection. After successful modeling validation, rats were randomized into the DKD group and the HucMSC-sEVs group, with normal rats as controls. At 8 weeks, HucMSC-sEVs were injected via the tail vein. After 24 weeks, kidney tissues were collected. Renal histopathology was assessed by HE staining, while Masson staining was employed to evaluate collagen deposition. The expression levels of necroptosis markers (p-RIPK1/RIPK1, p-RIPK3/RIPK3, and p-MLKL/MLKL) were quantified using qRT-PCR, Western blotting, and immunohistochemical staining. NRK-52E cells were treated with high glucose and HucMSC-sEVs. Transmission electron microscopy was used to observe necrotic phenomena in NRK-52E cells, and the expression levels of necroptosis marker proteins such as p-RIPK1/RIPK1, p-RIPK3/RIPK3, and p-MLKL/MLKL were detected by Western blotting and qRT-PCR. Results: Compared with the control group, the glomerular basement membrane in the DKD group rats was significantly thickened, and the interstitium exhibited fibrotic characterization, with a marked increase in the expression of necroptosis marker proteins. After treatment with HucMSC-sEVs, the pathological damage and fibrosis degree of the rat kidney tissues were alleviated, and the expression of necroptosis execution proteins was reduced. In vitro experiments revealed that high glucose-treated NRK-52E cells exhibited necrotic features including mitochondrial cristae fragmentation, endoplasmic reticulum dilation, and elevated expression of necroptosis markers (p-RIPK1/RIPK1, p-RIPK3/RIPK3, and p-MLKL/MLKL). Treatment with HucMSC-sEVs significantly attenuated these cellular alterations and reduced the expression levels of necroptosis-related proteins. Conclusion: HucMSC-sEVs can improve renal function and delay the progression of DKD in rats, and the mechanism may be attributed to the inhibition of necroptosis of renal tubular epithelial cells.

Objective: To integrate the two elements of selenium and magnesium into nanoparticles to prepare selenium-magnesium nanoparticles, and explore their antibacterial effect and antibacterial mechanism. Methods: The morphology and particle size distribution of selenium-magnesium nanoparticles were characterized by transmission electron microscopy and nanoparticle size analyzer. The antibacterial effect of selenium-magnesium nanoparticles was evaluated by plate counting method, disk diffusion method, and bacterial live/dead staining method. The content of reactive oxygen species in bacteria after exposure to nanoparticles was detected by DCFH-DA fluorescence probe to explore its potential antibacterial mechanism. Results: Selenium-magnesium nanoparticles are irregular spherical under transmission electron microscopy, and the particle size is mainly concentrated in the range of 50-70 nm. They exhibit excellent antibacterial properties, especially against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus. The potential antibacterial mechanism involves induction of reactive oxygen species production in bacteria. Conclusion: The prepared selenium-magnesium nanoparticles exhibit good antibacterial properties, it has been preliminarily demonstrated that bacterial killing is achieved by inducing the production of reactive oxygen species within bacterial.

Objective: To explore the disrupting effects of low-dose methyl-2-benzimidazole carbamate (CBZ) on glycolipid metabolism in mice during pubertal development period and its mechanism. Methods: A developmental (4-week-old) and adult (8-week-old) ICR mouse model with low-dose CBZ exposure was established. The mice were randomly divided into a blank control group (administered with corn oil) and different dose CBZ exposure groups (0.3, 3, 30 and 60 mg/kg ), and were given intragastric administration for 28 days. Commercial kits were used to detect the serum levels of total cholesterol (TC), total triglyceride (TG), low density lipoprotein cholesterol (LDLC), high density lipoprotein cholesterol (HDLC), creatinine (CRE), blood urea nitrogen (BUN), aspartate aminotransferase (AST), alanine aminotransferase (ALT), as well as the activity of glucose-6-phosphate dehydrogenase (G6PD) in the liver. Fasting blood glucose (FBG) was measured by a blood glucose meter. Body weight was monitored by weighing. The expression levels of genes related to glycolipid metabolism and endoplasmic reticulum stress pathway were detected by quantitative realtime PCR. Results: Compared with the blank control group, CBZ exposure significantly increased the serum lipid levels and various blood biochemical indexes in developing and adult mice (P<0.05 or P<0.01), and only significantly increased the FBG level and decreased the body weightof adult mice (P<0.05). CBZ exposure upregulated the expression of lipid synthesis and transport genes such as CD36 and SREBP-1, down-regulated the expression of lipid metabolism genes such as PPARα and HNF4α, as well as gluconeogenesis and glycogen synthesis genes such as G6PC, PEPCK and GSK3β in developing and adult mice(P<0.05). Meanwhile, CBZ up-regulated the expression of GRP78, PERK, CHOP, Bax, Caspase3 and other genes (P<0.05), down-regulated the expression of ATF4 and Bcl-2 genes (P<0.05), and activated the endoplasmic reticulum stress and apoptotic pathways. Conclusion: CBZ exerts endocrine and metabolic toxicity to developing mice, which can interfere with glycolipid metabolism and cause metabolic dysfunction by affecting the expression of glycolipid metabolism-related genes and activating the endoplasmic reticulum stress signaling pathway.

Objective: To assess the predictive value of systemic inflammatory markers such as C-reactive protein (CRP), and total bilirubin (TBil) combined with carbohydrate antigen 199 (CA199) for malignant biliary stricture (MBS). Methods: A total of 241 patients diagnosed with biliary strictures by endoscopic retrograde cholangiopancreatography in the Affiliated People′s Hospital of Jiangsu University from January 2019 to October 2022 were selected. According to the clinical diagnosis, the patients were divided into MBS group and benign biliary strictures (BBS) group. The relevant laboratory examination indicators of the two group patients were collected; and the clinicopathological characteristics of the two groups were compared. The risk factors of MBS were determined through binary Logistic regression analysis, and the predictive effects of each index were evaluated by using the receiver operating characteristic (ROC) curve. Results: Compared with the BBS group, patients in the MBS group were significantly older (P<0.05), and the levels of TBil, CA199, and platelet-lymphocyte ratio were greatly increased (P<0.05), while the levels of CRP and lymphocyte-monocyte ratio were significantly decreased (P<0.05). The results of multivariate Logistic regression analysis showed that advanced age, high CA199, high TBil and low CRP were all independent risk factors of MBS (P<0.05). The results of the ROC curve showed that the areas under the curve (AUC) of age, TBil, CA199 and CRP for predicting the occurrence of MBS were 0.574, 0.731, 0.750 and 0.523, respectively, and the AUC of age+CA199+TBil+CRP for predicting MBS was 0.807. Conclusion: Age, CRP and TBil combined with CA199 may be used to predict for MBS.

Objective: To investigate the correlation between the characteristic parameters of brown adipose tissue (BAT) and coronary artery calcification in 18F-FDG PET/CT images. Methods: A total of 2 399 patients who underwent 18F-FDG PET/CT examination at the Affiliated Hospital of Jiangsu University from December 2013 to April 2024 were collected. After 1∶1 propensity score matching (PSM), the patients were divided into BAT positive group (n=171) and BAT negative group (n=171) according to whether BAT was positive. Differences in general clinical data and the incidence of coronary artery calcification between the two groups were analyzed. Differences in BAT imaging characteristic parameters among six anatomical sites (the neck, supraclavicular fossa, axilla, mediastinum, paravertebral region, and abdomen) in patients of the BAT-positive group were analyzed, and the site with the highest BAT activity (the paravertebral region) was screened out. Furthermore, differences in paravertebral BAT imaging characteristic parameters among groups with different numbers of coronary artery calcification-involved branches and different severities of coronary artery calcification were analyzed. Results: After PSM, the incidence of coronary artery calcification and the coronary artery calcium score in the BAT positive group were significantly lower than those in the BAT negative group. In the BAT positive group, the SUVmax of paravertebral BAT was the highest among the six anatomical sites, and the difference was statistically significant compared with the SUVmax of abdominal and axillary BAT (all P<0.001). The SUVmean of paravertebral BAT significantly differed from that of axillary BAT (P<0.001), but showed no significant differences with other anatomical sites (all P>0.05). Both the volume and surface area of paravertebral BAT demonstrated statistically significant differences compared to abdominal and axillary BAT (all P<0.001). As both the number of coronary artery branches involved and the severity of calcification increased, all characteristic parameters of paravertebral BAT progressively decreased (all P<0.01). Conclusion: The presence of BAT is associated with a lower risk of coronary artery calcification. Paravertebral BAT may have a potential inhibitory effect on the occurrence and development of coronary artery calcification.

Objective: To explore the effect of crocin on the migration and invasion ability of anaplastic thyroid carcinoma (ATC) cells and its molecular mechanism of regulating epithelial-mesenchymal transition (EMT) via the Smad-dependent signaling pathway. Methods: ATC cells were treated with different concentrations of crocin. Cell proliferation and apoptosis were evaluated by CCK-8 assay and flow cytometry, respectively. The effects of crocin on ATC cell migration and invasion were detected by Transwell assays. Western blotting analysis was used to examine the expression of EMT markers (E-cadherin, N-cadherin, vimentin, fibronectin) and Smad signaling pathway-related proteins. A subcutaneous tumor model was established using BHT-101 cells to evaluate the inhibitory effect of crocin on tumor invasiveness. Immunohistochemistry and Western blotting were performed to detect the expression changes of MMP-2, MMP-9, N-cadherin and vimentin in tumor tissues. Results: At concentrations below 40 μmol/L, crocin treatment significantly reduced cell invasion and migration compared with the untreated group (P<0.01), accompanied by increased E-cadherin expression and decreased N-cadherin, vimentin and fibronectin expression. The inhibitory effect of crocin on Smad2/3 phosphorylation was alleviated after exogenous TGF-β intervention. In vivo experiments showed that crocin notably downregulated the expression of MMP-2, MMP-9, N-cadherin and vimentin in BHT-101 subcutaneous xenograft tumors. Conclusion: Crocin can inhibit the invasion and EMT of ATC cells by modulating the Smad-dependent signaling pathway.

Objective: To explore the effects of recombinant avirulent Newcastle disease virus La Sota strain expressing rabies virus glycoprotein (rL-RVG) on the polarization of murine macrophage RAW264.7 cells and to analyze the effect of polarized RAW264.7 cells on the viability of Lewis lung cancer cells. Methods: Murine macrophage RAW264.7 cells were divided into four groups: blank control group and rL-RVG groups at different multiplicities of infection(0.5, 1, 5 MOI); and the viability of RAW264.7 cells was assessed by using the CCK-8 assay. RAW264.7 cells were divided into three groups: RAW264.7-M0 type (complete media), RAW264.7-M1 type (induced by 1 μg/mL lipopolysaccharide +50 ng/mL IFN-γ), and RAW264.7-M2 type (induced by 20 ng/mL IL-4); and the morphological characteristics of these phenotypes were observed under phase-contrast microscopy. The protein expression levels of CD86 and Arg-1 were measured by immunofluorescence in four groups: Raw264.7-M1, Raw264.7-M2, rL-RVG+Raw264.7-M0, and rL-RVG+Raw264.7-M2. The mRNA expression levels of cytokines IL-4, IL-6, IL-10 and tumor necrosis factor-α (TNF-α) in the blank control group and rL-RVG groups were also assessed by qRT-PCR. Lewis lung cancer cells were co-cultured with conditioned media from the above groups and divided into four groups: blank control group and rL-RVG groups at different MOIs (0.5, 1, 5 MOI). The viability of Lewis lung cancer cells was evaluated by using the CCK-8 assay. Results: Compared with the blank control group, the cell proliferation rates in the 1, 5 MOI rL-RVG groups were significantly increased (both P<0.01). The morphological characteristics of Raw264.7-M0, M1 and M2 macrophages were round or oval, flat or irregular, and fibrous, elongated or spindle-shaped, respectively. Compared with RAW264.7-M0 type macrophages, the relative expression level of CD86 mRNA in RAW264.7-M1 type was significantly increased (P<0.01), and the relative expression level of Arg-1 mRNA in RAW264.7-M2 type was greatly increased (P<0.01). Compared with Raw264.7-M1 type, the relative fluorescence expression level of CD86 in rL-RVG+Raw264.7-M0 type was significantly increased (P<0.05); compared with Raw264.7-M2 type, the relative fluorescence expression level of CD86 in rL-RVG+Raw264.7-M2 type was significantly increased (P<0.05), and the relative fluorescence expression level of Arg-1 was significantly decreased (P<0.05). rL-RVG treatment led to an increase in the mRNA expressions of TNF-α and IL-6, while the mRNA expressions of IL-4 and IL-10 decreased. Compared with the blank control group, the viability of Lewis lung cancer cells in the 0.5, 1, and 5 MOI rL-RVG groups were significantly decreased (all P<0.01). Conclusion: rL-RVG could induce the polarization of RAW264.7 macrophages towards the M1 phenotype and subsequently inhibit the viability of Lewis lung cancer cells at MOIs ranging from 0.5 to 5.

Objective: To investigate the median effective dose (ED50) and 95% effective dose (ED95) of ciprofol for combined procedural sedation during spinal anesthesia in patients with different ages. Methods: A total of 67 patients scheduled for lower limb orthopedic surgery under subarachnoid block from December 2023 to March 2024 were enrolled and divided into three groups by age: the youth group (18-40 years old, 22 cases), the middleaged group (41-64 years old, 24 cases), and the elderly group (65-80 years old, 21 cases). After completion of spinal anesthesia, the Dixon up-and-down sequential method was used to administer ciprofol for procedural sedation, with an initial dose of 0.2 mg/kg and a dose gradient of 0.05 mg/kg between consecutive patients. Two minutes after administration, a modified observer′s assessment of alertness/sedation scale (MOAA/S) score ≤3 and a bispectral index (BIS) <85 were defined as satisfactory sedation. The dose for subsequent patients was adjusted, and the study was terminated after MOAA/S score ≥7 cross inflection points appeared. The Probit regression analysis was used to calculate ED50 and ED95 in each group. Vital signs, MOAA/S scores, BIS values, and the incidence of adverse reactions were recorded. Results: The ED50 and ED95 of the youth group were 0.263 mg/kg (95%CI: 0.232-0.300 mg/kg) and 0.318 mg/kg (95%CI: 0.288-0.509 mg/kg), respectively; those of the middle-aged group were 0.208 mg/kg (95%CI: 0.178-0.238 mg/kg) and 0.264 mg/kg (95%CI: 0.235-0.417 mg/kg); those of the elderly group were 0.178 mg/kg (95%CI: 0.130-0.217 mg/kg) and 0244 mg/kg (95%CI: 0.209-0.526 mg/kg). There were no statistically significant differences in vital signs, MOAA/S scores, BIS values, or the incidence of adverse reactions among the three groups during procedural sedation (all P>0.05). Conclusion: The effective doses of ciprofol for combined procedural sedation during spinal anesthesia depend on patients, age groups, and ED50 and ED95 decrease with increasing age.

Objective: To investigate the effects of extracellular matrix (ECM) protein SVEP1 on lipolysis and thermogenesis in adipocytes. Methods: The RNA-seq database of abdominal subcutaneous adipose tissue (SAT) of 236 normal and obese individuals constructed previously was used to analyze the correlation between SVEP1 expression level and body mass index (BMI), fat mass, fat percentage and waist circumference. 3T3-L1 preadipocytes and human stromal vascular fraction (SVF) cells were differentiated into mature adipocytes respectively. The expression of Svep1 in adipocytes stimulated by rosiglitazone and Forskolin was detected by realtime quantitative PCR (qRT-PCR). Based on the Pearson correlation coefficient (r>0.4, P<0.05) between RPKM values of SVEP1 and other genes in the RNA-seq database of human SAT, co-expressed gene set was screened and KEGG enrichment analysis was performed. Svep1 was knocked down in mouse primary adipocytes by lentivirus-mediated shRNA. qRT-PCR and Western blotting were used to assess the expression of thermogenic genes and proteins in adipocytes. Glycerol detection kit was used to detect the level of lipolysis stimulated by isoproterenol (ISO), and Western blotting was used to detect the phosphorylation level of lipolysis-related proteins. Results: The expression level of SVEP1 in human abdominal SAT was positively correlated with obesity related indicators (BMI, fat mass, fat percentage and waist circumference). Cold stimulation could down-regulate Svep1 transcription level in brown and beige adipocytes in mice. The transcription level of Svep1 was down-regulated in mature 3T3-L1 adipocytes and human adipocytes when stimulated by Rosi or Forskolin. KEGG enrichment analysis showed that Svep1 co-expressed gene set was enriched in the regulation of lipolysis and insulin resistance related pathways. Svep1 knockdown could significantly promote ISO-stimulated lipolysis in mouse white adipocytes and increase the expression of thermogenic genes (Ucp1, Dio2, Pgc1α, Elovl3 and Cidea) in mouse beige adipocytes. Conclusion: This study demonstrated an association between Svep1 expression and adipocyte function, and it may be implicated that Svep1 might be involved in obesity-related adipose tissue dysfunction and contribute to the development and progression of obesity.

Objective: To investigate the risk factors for diuretic resistance in patients with acute exacerbation of chronic heart failure and to construct a Nomogram model, aiming to provide a reference for the early identification of patients with diuretic resistance. Methods: A retrospective study was designed by including 215 patients with acute exacerbation of chronic heart failure who were admitted to the Affiliated People′s Hospital of Jiangsu University from January 2021 to August 2024. Patients were grouped into two groups, one is resistance group, the other non-resistance based on the criteria for diagnosing diuretic resistance, and baseline data were collected for both groups. Using the least absolute shrinkage and selection operator (LASSO) regression for univariate screening, followed by multifactorial Logistic regression for further screening, a nomogram model was constructed and subjected to internal validation. Results: LASSO regression selected 6 variables, and further analysis using multiple Logistic regression revealed that an elevated Nterminal pro B-type natriuretic peptide (NT-proBNP) level was an independent risk factor for diuretic resistance ［OR (95%CI): 2.342 (1.087-5.043), P<0.05］, while higher levels of estimated glomerular filtration rate (eGFR), hemoglobin, serum sodium, and left ventricular ejection fraction (LVEF) were identified as protective factors ［OR (95%CI): 0.978 (0.965-0.990), 0.973 (0.958-0.989), 0.893 (0.822-0.971), 0.944 (0.918-0.971), all P<0.05］. A nomogram model was constructed based on the above risk factors. The area under the receiver operating characteristic (ROC) curve was 0.846 (95%CI: 0.795-0.897). The calibration curve and Hosmer-Lemeshow test indicated that the model fit well (P=0.159). The clinical decision curve analysis demonstrated that the model provided a favorable net clinical benefit when the threshold probability ranged from 0% to 87.0%. Conclusion: The Nomogram model, incorporating NT-proBNP, hemoglobin, blood sodium, eGFR, and LVEF, effectively assesses the risk of diuretic resistance in patients with acute exacerbation of chronic heart failure.

Objective: To explore the feasibility of preoperative prediction of perineural invasion (PNI) and lymphovascular invasion (LVI) in advanced gastric cancer using dual-energy computed tomography (DECT) venous phase imaging features and spectral parameters of a hypotonic water-filled stomach, along with clinical laboratory information. Methods: A retrospective analysis was conducted on 161 cases of advanced gastric cancer that underwent DECT imaging within one week before surgery, and the cases were randomly divided into training sets and test sets in a 7∶3 ratio. Based on postoperative pathological assessment, 115 cases demonstrated LVI and/or PNI positivity, whereas 46 cases were negative for both. A predictive model for LVI/PNI was developed using venous phase imaging of a hypotonic water-filled stomach, DECT parameters ［including the slope of the spectral Hounsfeld unit curve (between 40 keV and 100 keV), normalized iodine concentration (NIC), and effective atomic number］, and clinical laboratory data (inflammatory and tumor markers). The predictive performance of the model was evaluated using the area under the ROC curve (AUC), and its clinical utility was assessed using decision curve analysis. Results: The AUC values of the radiomics model (Rad-score) in the training sets and test sets were 0.776 (95%CI: 0.653-0.821) and 0.781（95%CI：0.582~0.847）, respectively. The independent predictors for the DECT parametric model was NIC, with AUC values of 0.729（95%CI：0.615~0.790 in the training sets and 0.771（95%CI：0.604~0.864）in the test sets. For the clinical information predictive model, the independent predictor was lymphocyte percentage, with AUC values of 0.693 (95%CI: 0.638-0.805) in the training sets and 0.502 (95%CI: 0.352-0.648) in the test sets. The combined model integrating the Rad-score, DECT parameters, and clinical information had independent predictors including Rad-score, NIC, and lymphocyte percentage. The AUC values for this combined model were 0.880 (95%CI: 0.701-0.859) in the training sets and 0830 (95%CI: 0.602-0.857) in the test sets, demonstrating superior performance compared to the radiomics model, DECT parametric model, and clinical model. The DeLong test showed that the AUC of the combined model was significantly higher than that of the radiomics model, DECT parametric model, and clinical information model in the training sets (Z=1.979, P=0.048; Z=3.199, P=0.001; Z=3.053, P=0.001). In the test sets, the AUC of the combined model was also significantly higher than that of the clinical information model (Z=2.417, P=0.015). Decision curve analysis revealed that when the risk threshold ranges from 0.15 to 0.96, adopting the combined model for treatment guidance yielded a higher clinical net benefit rate. Conclusion: The integrated model, incorporating radiomics, NIC, and lymphocyte percentage, serves as a comprehensive predictive model for assessing lymphovascular invasion and perineural invasion status in advanced gastric cancer.

Objective: To investigate the association between the polymorphisms of meteorinlike protein (METRNL) gene rs4986080 and type 2 diabetes mellitus (T2DM). Methods: A total of 467 subjects were selected, including 315 T2DM patients and 152 individuals with normal glucose tolerance (NGT). The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the polymorphisms of METRNL gene rs4986080 in all subjects, then analyzed the association between these polymorphisms and T2DM. Results: The constituent ratio of GG genotype and the frequency of G allele at rs4986080 locus of METRNL gene was higher in the T2DM group compared to the NGT group (P<0.05). Compared to individuals with GG genotype at the METRNL gene rs4986080 locus, those with AA+AG genotypes showed significant lower waisttohip ratio, fasting plasma glucose (FPG), 2hour postprandial glucose (2hPG), glycated hemoglobin (HbA1c), and homeostasis model assessment of insulin resistance (HOMA-IR) (P<0.05). Logistic regression analysis showed that A allele at rs4986080 locus of METRNL gene was probably a protective factor for T2DM. Conclusion: The polymorphisms of METRNL gene rs4986080 may be associated with the occurrence of T2DM, and A allele may be a protective factor for T2DM.

Objective: To analyze the expression levels of gap junction protein beta 5 (GJB5) in pancreatic cancer and its relationship with patient prognosis, and to preliminarily explore its effect on the malignant biological behavior of pancreatic cancer cells. Methods: Using data from the UCSC XENA database, the relative expression level of GJB5 mRNA in human pan-cancer tissues and their correlation with patient prognosis and various clinicopathological characteristics were analyzed. Gene ontology (GO) and Kyoto encyclopedia of gene and genomes (KEGG) analysis in over representation analysis were used to explore the potential biological functions of GJB5. Gene set enrichment analysis (GSEA) was employed to predict the signaling pathways regulated by GJB5related differentially expressed genes. qRTPCR was used to detect the expression levels of GJB5 mRNA in human pancreatic ductal epithelial cells (HPNE) and five human pancreatic cancer cell lines (CFPAC1, PaTu 8988T, BxPC3, MIA PaCa2 and PANC1), and the optimal pancreatic cancer cell lines were selected for transfection. In vitro experiments, lentiviral transfection was used to establish GJB5 knockdown and overexpression models in human pancreatic cancer PaTu 8988-T and BxPC-3 cells, and to assess the effects of GJB5 on the growth, proliferation, migration, and invasion of pancreatic cancer cells. Results: Bioinformatics analysis showed that the relative expression of GJB5 mRNA was significantly upregulated in pancreatic cancer tissues compared to adjacent normal tissues (P<0.05). High GJB5 mRNA expression levels were significantly correlated with the poor prognosis of pancreatic cancer patients (P<0.05). The expression of GJB5 mRNA was significantly correlated with histological grade of pancreatic cancer patients (P<005)，and there was no significant correlation with gender, smoking history, alcohol history, T stage and N stage (P>005). GO/KEGG analysis indicated that GJB5 plays an important role in various biological processes, including signal release, synaptic signaling regulation, chemical synapse transmission regulation, membrane potential regulation, and hormone transport. GSEA results suggested that GJB5-related differentially expressed genes might be involved in regulating multiple signaling pathways, such as KRAS, apoptosis and protein ubiquitination. Compared with HPNE cells, the relative expression of GJB5 mRNA was significantly increased in the five human pancreatic cancer cell lines (all P<0.05). Knockdown of GJB5 mRNA significantly inhibited the growth, proliferation of PaTu 8988-T and BxPC-3 cells, migration and invasion capabilities of PaTu 8988T cells; on the contrary, overexpression of GJB5 mRNA promoted the growth, proliferation of PaTu 8988-T and BxPC-3 cells, migration and invasion capabilities of PaTu 8988-T cells. Conclusion: GJB5 is significantly overexpressed in pancreatic cancer tissues and pancreatic cancer cell lines, and GJB5 overexpression is related to the poor prognosis of pancreatic cancer patients, and promotes the malignant growth of pancreatic cancer cells lines PaTu 8988-T and BxPC-3.

Objective: To investigate the role of the long noncoding RNA (lncRNA) NONRATT021203.2 in the progression of cancer-induced bone pain (CIBP) and its potential molecular mechanisms. Methods: A CIBP model was constructed in Sprague-Dawley (SD) rats by intratibial inoculation of Walker 256 breast cancer cells. Sixteen adult female SD mice weighing 180-200 g were randomly divided into control group and CIBP group, with 8 rats in each group, and injected into tibiae with 10 μL normal saline and 10 μL Walker 256 breast cancer cell suspension (1×10⁸ cells/mL), respectively. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) of the modeled lower limb of rats were detected by behavioral analysis. The expression levels of lncRNA NONRATT021203.2 and miRNA-138-5p in DRG were measured by using quantitative real-time PCR (qRT-PCR), and the localization of miRNA-138-5p in DRG tissue was examined by fluorescence in situ hybridization (FISH). Seven days after modeling, 10 rats with CIBP were selected and divided into CIBP+siRNA group (n=5) and CIBP+siNC group (n=5), lncRNA NONRATT021203.2-siRNA and siNC were injected intrathecally, respectively. The relative expression levels of lncRNA NONRATT021203.2 and miRNA-138-5p in the two groups were detected by qRT-PCR. Additionally, seven days after modeling, 16 rats with CIBP were divided into CIBP+mimics (n=8) and CIBP+NC group (n=8), miRNA-138-5p mimics and negative control oligonucleotide were injected intrathecally, respectively. PWT and PWL were detected by behavioral test. Finally, the binding sites of lncRNA NONRATT021203.2 and miRNA-138-5p were analysed by using miRand and miRwalk3.0 software. Results: Compared with control group, CIBP group showed significantly reduced PWT and PWL (P<0.05), markedly increased expression of lncRNA NONRATT021203.2 (P<0.01), and significantly decreased expression of miRNA-138-5p (P<0.001). FISH results showed that miRNA-138-5p was localized in neuronal cytoplasm. Compared with CIBP+siNC group, CIBP+siRNA group exhibited significantly reduced lncRNA NONRATT021203.2 expression (P<0.05) and increased miRNA-138-5p expression (P<0.05). Compared with CIBP+NC group, PWT was significantly increased in CIBP+miRNA-138-5p mimics group (P<0.01). Bioinformatics analysis revealed the presence of a potential binding site between lncRNA NONRATT021203.2 and miRNA-138-5p. Conclusion: LncRNA NONRATT021203.2 could reduce the intracellular free miRNA-138-5p level by competitively binding to miRNA-138-5p, thereby promoting the occurrence and development of bone cancer pain.

Objective: To prepare shikonin liposomes (SHK-L) and preliminarily evaluate their anti-aging effects. Methods: The SHK-L prescription was optimized through single factor experiments to investigate its in vitro release and oral bioavailability in rats. The in vivo safety was evaluated through zebrafish toxicity experiments, and the anti-aging effect was examined by staining the β-galactosidase in aging zebrafish. Results: The optimal prescription was SHK 6 mg, lecithin 70 mg, cholesterol 10 mg, vitamin E polyethylene glycol succinate (TPGS) 30 mg, and sodium dodecyl sulphate (SDS) 1.5 mg, with a particle size of (206.96±0.75) nm, polydispersity coefficient (0.186±0.030), encapsulation rate of (96.88±2.19)% and drug loading was (5.36±0.01)%. SHK-L improved the drug release rate compared to SHK. According to in vivo pharmacokinetic data, SHK-L had a relative bioavailability of 413.37% of SHK. At 96 hours post-administration, LC₅₀ of shikonin and SHK-L in zebrafish were (0.112±0.007) μg/mL and (0.141±0.012) μg/mL, respectively. Conclusion: SHK-L can enhance the solubility and oral bioavailability of insoluble drugs, as well as their anti-aging effect and safety.

Objective: To develop a wound dressing incorporating Clostridium butyricum (C. butyricum, anaerobic probiotic), which is capable of producing antibacterial substances and exerting hydrogen-producing properties under normoxic conditions, and investigate the effects of its selective bacteria inhibitionand reactive oxygen species (ROS) scavenging. Methods: C. butyricum and Bacillus subtilis (B. subtilis, aerobic probiotic) were simultaneously encapsulated in hydrogel to prepare a dual-probiotic hydrogel (BC-gel). B. subtilis was used to consume oxygen within the hydrogel. This creates an anaerobic microenvironment for C. butyricum to maintain its viability. The selective bacteria inhibition of BC-gel was evaluated by plate counting method, while the selective scavenging of ROS was evaluated through flow cytometry. Results: BC-gel can effectively inhibit the proliferation of Escherichia coli and methicillin-resistant Staphylococcus aureus. While maintaining the normal growth of Bifidobacterium and Lactobacillus, BC-gel has the ability to scavenge ·OH and ONOO- free radicals, but has no significant decreasing effect on O-2· and NO·. Conclusion: BC-gel exhibits favorable selective bacteria inhibition and ROS scavenging properties.