To determine the expression level of TG-interacting factor 1 (TGIF1) in gastric cancer (GC) and its correlation with prognosis, and investigate the characteristics of immune microenvironment of TGIF1-overexpressing GCs and its clinical significance.

Integrated analysis was performed using transcriptomic data from The Cancer Genome Atlas (TCGA), Asian Cancer Research Group (ACRG), and our in-house GC transcriptome database. Immunohistochemistry (IHC) assay was employed to compare TGIF1 expression between GC and normal gastric mucosa tissues. Based on Kaplan-Meier Plotter online database, the correlation between TGIF1 expression and clinical prognosis was evaluated. Transcriptomic data were analyzed to identify functional enrichment features of GC with high TGIF1 expression. The GENIE3 toolkit and STRING database were utilized to predict TGIF1-regulated target genes and protein-protein interaction (PPI) networks, respectively to explore the potential immune-related signaling pathways regulated by TGIF1. Single-cell RNA sequencing (scRNA-seq) was applied to analyze the association between TGIF1 expression and tumor microenvironment (TME) disorder.

Transcriptomic analysis revealed significantly higher TGIF1 expression in GC tissues compared to normal tissues (P<0.01). Patients with high TGIF1 expression exhibited poorer clinical prognosis (P<0.05). IHC assay confirmed elevated TGIF1 expression in GC tissues than normal tissues (P<0.01). GC with high TGIF1 expression was enriched in immune regulatory pathways, including immunosuppressive cytokines such as chemokine C-C motif ligand 20 (CCL20). These tumors displayed an immunosuppressive TME, characterized by abundant immunosuppressive NK cells, mast cells, regulatory T cells (Tregs), myeloid cells, and exhausted CD8⁺ T cells. Cell interaction analysis suggested that TGIF1-overexpressing GC cells may engage with Tregs via the CCL20-CCR6 axis. Co-high expression of signature gene sets from these interacting cells was associated with significantly shorter survival in the patients (P<0.05).

TGIF1 is highly expressed in GC tissues and may serve as a biomarker for immunosuppressive TME and poor prognosis. TGIF1-overexpressing GC cells may interact with multiple immune cells, particularly Tregs, to induce an immunosuppressive microenvironment.

To elucidate the effects of miR-616 on the malignant biological processes of osteosarcoma and to preliminarily explore its potential mechanisms.

In situ hybridization (ISH) was employed to analyze miR-616 expression in 11 paraffin-embedded osteosarcoma specimens collected in our department during January 2018 to December 2019. Quantitative real-time PCR (qRT-PCR) was used to compare the mRNA expression level of miR-616 in the osteoblast cell line hFOB1.19 and osteosarcoma cell lines 143B and HOS. Stable cell lines with miR-616 knockdown or overexpression were established via lentiviral transfection in 143B and HOS cells. Cell proliferation and apoptosis were detected by flow cytometry, while cell invasion and migration were assessed using Transwell and colony formation assays, respectively. To evaluate the effect of miR-616 on tumor growth in vivo, 10 female nude mice (4 weeks old, weighing 18~20 g) were randomized into a control group and a miR-616 overexpression group. After the xenograft tumor model was constructed, the growth of subcutaneous tumors was monitored. Finally, next-generation sequencing and a dual-luciferase reporter assay were performed to identify the target genes of miR-616.

ISH results showed that miR-616 expression was up-regulated in osteosarcoma tissues than adjacent tissues, and primarily localized in the cytoplasm. qRT-PCR confirmed that miR-616 level was significantly higher in 143B and HOS cells than hFOB1.19 cells (P<0.05). In vitro experiments revealed that miR-616 overexpression enhanced the proliferation, migration and invasion, while suppressing apoptosis in 143B and HOS cells (P<0.01). Conversely, miR-616 knockdown weakened these malignant phenotypes (P<0.05), with miR-616-3p showing a stronger effect on apoptosis than miR-616-5p. Animal experiments demonstrated that the tumor weight in the miR-616 overexpression group was significantly greater than that of the control group (98.00±17.22 vs 33.60±8.08 mg, P<0.01). Furthermore, KLF2 was identified and confirmed as a direct target of miR-616.

MiR-616 promotes malignant biological behaviors in osteosarcoma, and its expression level indicates that it may serve as a potential therapeutic target.

To investigate whether microRNA-21-5p (miR-21) plays a protective role in hyperoxia-induced acute lung injury (HALI) by regulating ferroptosis through the STAT3/P53/SLC7A11 axis.

The interaction between STAT3 and P53 was analyzed by co-immunoprecipitation (Co-IP). Fifty 9-week-old male C57BL/6J mice were randomly divided into normoxia (Control) group, HALI group, miR-21 overexpression (miR-21) group, STAT3 inhibitor (HY-13818) group, and ferrostatin-1 (Fer-1) group. After the mice from the miR-21 group received miR-21-5p AAV6 or empty vector via tracheal catheter instillation, the animals were then monitored for 3 weeks. The HY-13818 group was intraperitoneally injected with HY-13818 (10 mg/kg) 3 times weekly for 2 weeks. The Fer-1 group received 0.8 mg/kg ferrostatin-1 via tail vein injection once daily for 2 consecutive days during modeling. The HALI model was established by exposure to >90% oxygen. After 48 h of hyperoxia, blood samples were collected via orbital sampling for RT-PCR analysis of miR-21 expression. Lung tissues were harvested for wet/dry weight ratio and assessment of histopathological changes via HE staining for lung injury score. Activity of superoxide dismutase (SOD) and contents of malondialdehyde (MDA), Fe²⁺, glutathione (GSH), and reactive oxygen species (ROS) were measured using photocolorimetry, spectrophotometry and fluorometry. Western blotting was used to evaluate the protein expression of STAT3, P53, SLC7A11, and GPX4.

The results of Co-IP showed that STAT3 could bind to P53. The HALI group exhibited obviously destroyed alveolar structure, disordered arrangement, thickened interval, with a large number of infiltrated neutrophils and collapsed alveoli, and had significantly increased pathological score of lung injury and ratio of lung W_wet/D_dry weight when compared with the Control group (P<0.05). In the miR-21 group, HY-13818 group and Fer-1 group, the severity of lung injury was significantly reduced, and the pathological score of lung injury and the ratio of W_wet/D_dry weight were decreased (P<0.05) when compared with the HALI group. Compared with the control group, the contents of MDA, Fe²⁺ and ROS were increased (P<0.05), the activity of SOD and content of GSH were declined (P<0.05), the protein levels of STAT3 and P53 were increased (P<0.05), and those of SLC7A11 and GPX4 were decreased (P<0.05) in the HALI group. Compared with the HALI group, decreased MDA and ROS levels (P<0.05), enhanced SOD activity, Fe²⁺ and GSH levels (P<0.05), down-regulation of STAT3 and P53 (P<0.05) and up-regulation of SLC7A11 and GPX4 (P<0.05) were observed in the miR-21 group and HY-13818 group.

MiR-21 alleviates HALI, which may be related to its inhibition of ferroptosis through the STAT3/P53/SLC7A11 axis.

To investigate the effects of paeoniflorin (PF) on lipopolysaccharide (LPS)-induced aerobic glycolysis in renal tubular epithelial cell line HK-2 and its underlying mechanism of action.

This study consists of a preliminary experiment and a formal experiment. Preliminary experiment: CCK-8 assay and RT-qPCR were used respectively to measure cell viability and mRNA expression levels of inflammatory factors in HK-2 cells after LPS stimulation to determine the optimal LPS concentration for modeling as well as to evaluate the toxicity of PF and screen for its appropriate concentration. Formal experiment: HK-2 cells were divided into control group (CON group), LPS group, LPS+PF group and LPS+PF+740Y-P group. LPS was used to establish a cell model of sepsis associated-acute kidney injury (SA-AKI) in HK-2 cells, and then the cell model was treated with PF and PI3K activator 740Y-P, correspondingly for 24 h. CCK-8 assay was employed to detect cell viability, and Extracellular Acidification Rate (ECAR) Kit was utilized to measure the rate. The contents of IL-1β, IL-18, lactic acid (Lac) and lactate dehydrogenase A (LDHA) were determined with ELISA. Western blotting was applied to detect the expression of p-PI3K, p-AKT, HIF-1α, pyruvate kinase 2 (PKM2, a key enzyme in aerobic glycolysis) and NOD-like receptor thermal protein domain associated protein 3 (NLRP3), and immunofluorescence assay was performed to observe the expression and distribution of PKM2.

① Our preliminary experiment identified that the optimal concentration of LPS for modeling was 20.0 μg/mL, a safe dosage range of PF was 0~100.0 μmol/L, and its optimal therapeutic concentration was 25.0 μmol/L. ② Compared with the CON group, LPS stimulation resulted in significantly decreased cell viability (P<0.05), increased ECAR (P<0.05), elevated contents of IL-1β, IL-18, Lac and LDHA (P<0.05), up-regulated protein levels of p-PI3K, p-AKT, HIF-1α, p-PKM2 and NLRP3 (P<0.05), and enhanced fluorescence intensity of PKM2 in the nucleus of cells (P<0.05). Compared with the model group, PF treatment reversed all above effects induced by LPS stimulation (all P<0.05). Compared with the LPS+PF group, in the LPS+PF+740Y-P group, ECAR was elevated (P<0.05), the contents of IL-1β, IL-18, Lac and LDHA were increased (P<0.05), and the relative expression levels of p-PI3K, p-AKT, HIF-1α, p-PKM2 and NLRP3 were increased (P<0.05), and the fluorescence intensity of PKM2 was strengthened (P<0.05) and enhanced in the nucleus (P<0.05).

PF reduces aerobic glycolysis in HK-2 cells and alleviates the inflammatory response by inhibiting the PI3K/AKT/HIF-1α signaling pathway.

To investigate the protective effects of berberine (BBR) against drug-induced liver injury (DILI) in mice and the change profiles in gut microbiota, and to elucidate the underlying mechanism by which BBR alleviates DILI through modulation of the gut microbiota.

Acetaminophen (APAP) administration was performed on C57BL/6J mice with or without pre-treatment of ampicillin-neomycin-metronidazole-vancomycin (ABX, depletion of gut microbiota) to establish a DILI model. Based on body weight, 12 mice were randomly divided into an APAP group and a BBR group, with 6 animals in per group. Serum and liver tissue samples were collected from both groups to determine the effects of intragastrical administration of BBR on serum transaminase levels, inflammatory indicators, and liver histopathology. Fresh fecal samples were also collected to profile the gut microbiota using 16S rRNA gene sequencing. Then the alpha and beta diversity, as well as the overall microbial community structure, were compared between the 2 groups. Pearson correlation analysis was employed to analyze the relationships between differentially abundant bacteria and the serological indicators.

Intragastrical administration of BBR significantly reduced APAP-induced serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and ameliorated the proportion of hepatocellular necrosis caused by APAP. The results of 16S rRNA gene sequencing revealed that there were no statistical differences in the alpha diversity indices (Chao1 and Shannon) between the APAP-BBR and APAP groups, but a significant difference was observed in beta diversity (P<0.05). Compared to the APAP-BBR group, the relative abundances of Alloprevotella and Ruminococcaceae_UCG_014 were significantly increased in the APAP group (P<0.05). Conversely, the relative abundance of Parabacteroides was significantly higher in the APAP-BBR group than the APAP group (P<0.05). Pearson correlation analysis demonstrated that Alloprevotella was strongly correlated with AST level (P<0.05), while both Ruminococcaceae and Parabacteroides showed strong correlations with ALT and AST levels (P<0.05).

Intragastric administration of BBR can mitigate the progression of DILI in mice, potentially through modulating the gut microbiota.

To investigate the influencing factors for stroke-associated pneumonia (SAP) in acute ischemic stroke (AIS) patients after endovascular treatment (EVT).

A retrospective case-control trial was conducted on 426 AIS patients with large vessel occlusion (LVO) in anterior circulation admitted in the neurological departments from First Affiliated Hospital of Army Medical University and Zigong Third People’s Hospital during January 2017 and April 2021. Based on SAP occurrence or not, they were divided into an SAP group and a non-SAP group. Demographic information (gender and age), TOAST stroke subtypes (large artery atherosclerosis type, cardiac embolism type, others), vascular risk factors (hypertension, hyperlipidemia, diabetes, atrial fibrillation, smoking, prior stroke history, smoking), and post-onset clinical data [dysphagia, LDL cholesterol, white blood cells, neutrophils, baseline and postoperative NIHSS scores, endovascular outcomes (mTICI grade 2b or 3), 90-day good prognosis (mRS 0-1)] were collected and compared between the 2 groups. Multivariate logistic regression analysis was performed using the parameters with P<0.1 in univariate analysis as independent variables to investigate factors influencing SAP occurrence after EVT in AIS patients.

Among the 426 participants, SAP occurred in 194 cases (45.5%). Multivariate logistic regression analysis revealed that admission white blood cell count (OR=1.125, 95% CI: 1.043~1.213, P=0.000 2), postoperative NIHSS score (OR=1.019, 95% CI:1.001~1.037, P=0.041), and male (OR=1.687, 95% CI:1.078~2.638, P=0.022) were associated with SAP occurrence after EVT in AIS patients.

Higher admission white blood cell count, elevated postoperative NIHSS score, and male gender are risk factors for SAP in AIS patients after EVT. These risk factors should be focused on clinical practice to control SAP incidence.

To quantitatively evaluate age-, sex-, and segment-related characteristics of lumbar disc degeneration (LDD) in patients with low back pain (LBP) using T1rho MRI, and assess its diagnostic value in early-stage disc degeneration.

A retrospective cross-sectional study was conducted on 89 LBP patients (balanced distribution across ages 20~29, 30~39, 40~49, 50~59 years and genders) admitted in Department of Orthopedics of First Affiliated Hospital of Army Medical University from March 2024 to February 2025. All patients underwent lumbar MRI including T1rho sequences. Two senior radiologists performed blinded Pfirrmann grading on 445 intervertebral discs and measured T1rho values of the nucleus pulposus (NP). Kappa test was used to evaluate the inter-observer agreement of Pfirrmann grading, and Spearman correlation coefficient was employed to analyze the correlation between Pfirrmann grade and NP-T1rho value. Disc degeneration characteristics were compared across different ages, sexes, and spinal segments. After adjusting for body mass index (BMI) and gender, partial correlation analysis was used to assess the correlation between NP-T1rho value of each lumbar segment and age. Receiver operating characteristic (ROC) curve was plotted to analyze the diagnostic performance of T1rho quantification for early LDD in LBP patients.

Good inter-observer agreement was observed in Pfirrmann grading across segments, with a Kappa coefficient of >0.70. The NP-T1rho values of all NP were in a strong negative correlation with Pfirrmann grades (r=-0.877, P<0.001). The NP-T1rho values were progressively decreased from Pfirrmann gradesⅠ to Ⅳ (P<0.001), with a steep decline between gradesⅡ and Ⅲ. The patients aged 20~29 and 30~39 years showed significantly higher NP-T1rho values in Pfirrmann grades Ⅰ and Ⅱ discs when compared with those aged 40~49 and 50~59 years (P<0.05). The males had significantly higher mean NP-T1rho values in grade I than the females (P=0.006). In the upper spinal segments (L1~L4), the males exhibited significantly higher mean NP-T1rho value than the females (P=0.025), and NP-T1rho value was decreased obviously with increase of age across all age groups (P<0.05). Except L5/S1 segments, NP-T1rho value and age in all spinal segments had significant negative correlations (r=-0.548~-0.349, P<0.001). The area under the curve (AUC) for distinguishing grade Ⅰ from Ⅱ discs was 0.824 (95%CI: 0.772~0.876), and for grades Ⅱ from Ⅲ was 0.978 (95%CI: 0.964~0.992).

T1rho MRI provides accurate quantitative assessment of early lumbar disc degeneration in LBP patients, with an NP-T1rho value of 79.87 ms serving as a discriminative threshold between grade Ⅱ and grade Ⅲ.

To evaluate the application efficacy of structured symptom intervention program in patients with chronic kidney disease (CKD) during the peri-dialysis period.

A non-simultaneous control study was conducted on 151 peri-dialysis outpatients having not yet initiated dialysis and being followed up who were subjected with convenience sampling from Department of Nephrology of Second Affiliated Hospital of Army Medical University from April to September 2024. According to the time period of their visits, the patients who visited from April to June 2024 were assigned into a control group (n=75), and those from July to September 2024 into an experimental group (n=76). The control group received conventional symptom intervention (telephone symptom reporting + health education), while the experimental group received the intervention as the control group and a structured symptom intervention program covering 4 evidence-based modules: symptom identification, assessment, intervention, and outcome. The efficacy of above treatments was evaluated before and at 3 months after intervention. Dialysis symptom index was used to assess the degree of symptom distress and the number of symptoms. MOS 36-Item Short Form Health Survey (SF-36) was employed to evaluate the quality of life. The differences in clinical indicators and endpoint events were compared between the 2 groups after intervention.

The experimental group obtained more significant reduction in the total score of symptom distress than the control group (P=0.021). After intervention, the number of symptoms was decreased in both groups (P<0.001), but no statistical difference was observed between the groups. The score of mental health dimension in SF-36 was obviously improved in the experimental group (P=0.004), which was notably better than that in the control group (P=0.033). The experimental group exhibited significantly higher prealbumin level than the control group (P=0.019), and stable albumin level, which was significantly decreased in the control group (P=0.035). The incidence of endpoint events was remarkably lower in the experimental group than the control group (P=0.028).

The structured symptom intervention program implements intervention through a closed-loop symptom module, which can effectively alleviate the symptom distress of patients during the peri-dialysis period, improve mental health and reduce the short-term risk for endpoint events.

To explore the networked association among the dimensions of self-efficacy in patients with rheumatoid arthritis (RA) and to identify core efficacy and bridge efficacy, and provide a basis for formulating precise nursing intervention strategies.

A total of 652 RA patients admitted in our hospital from September 2024 to January 2025 were enrolled with convenience sampling. The general information questionnaire and Rheumatoid Arthritis Self-Efficacy Scale (RASE) were used for assessment. Exploratory factor analysis was used to extract efficacy symptom clusters. With aid of R project, network analysis was employed to construct an association network among efficacy dimensions to calculate centrality indicators (strength, closeness, betweenness) to identify core efficacy and bridge efficacy.

Exploratory factor analysis extracted 7 efficacy symptom clusters, with a cumulative variance contribution rate of 64.539% (P<0.001). Network analysis showed that the network density was 0.143, suggesting that there were moderate correlations among the self-efficacy dimensions. “Relaxation efficacy 1 (r1)” and “pain efficacy 1 (a1)” had the strongest correlation (r=0.73). Interpersonal efficacy 2 (i2) had the highest intensity centrality (6.88), and relaxation efficacy 3 (r3) had the highest tightness (0.0125), and medication efficacy 1 (m1) had the highest mediation (116), which were the core efficacy and bridge efficacy in this network group.

There are complex network-like correlations among the various dimensions of self-efficacy in RA patients. Interpersonal efficacy is the core driving factor, while relaxation and medication efficacies play the bridging role, jointly influencing the overall level of patients’ self-management ability.

To analyze the electrophysiological and genetic examination results of children with congenital stationary night blindness (CSNB) without fundus abnormalities.

A retrospective study was conducted on 11 children diagnosed with CSNB in our department from August 2019 to March 2025. Their full-field electroretinogram (ffERG) and the results of genetic testing were analyzed.

Among the 11 patients, there were 10 males and 1 female, at a mean age of 7.5±2.1 years. In the optometry, 1 patient had hyperopia and the other had myopia, and all of them obtained a corrected visual acuity of 0.48±0.11 after wearing glasses. In fundus photography examination, except for the leopard pattern changes in high myopia, no obvious abnormalities were found in the remaining fundus. In ffERG examination for all patients, the amplitude of the a-wave of scotopic ERG 3.0/10.0 was normal or slightly to moderately decreased, the amplitude of the b-wave was decreased significantly, and the amplitude ratio of the b-wave to the a-wave was <1, showing a negative-phase wave, which belonged to the CSNB Schubert-Bornschein waveform. The results of genetic test revealed that all the 11 patients carried pathogenic mutations, including 6 cases with CACNA1F mutations, 2 cases with GPR179 mutations, 2 cases with NYX mutations, and 1 case with GRM6 mutation.

For children with normal fundus manifestations but corrected visual acuity lower than normal after wearing glasses, ffERG examination should be carried out as soon as possible to screen for CSNB, and a diagnosis should be made in combination with genetic testing. This is helpful for evaluating the heredity and prognosis of the disease, increasing the chance of early detection, and reducing excessive amblyopia training and treatment.

To analyze the burden of pressure ulcers and its changing trends among the general population (all age groups) and the elderly aged 65 years and above in China based on the global burden of disease (GBD) data from 1990 to 2021, predict the burden trends from 2022 to 2035, and address the public health issues of its unclear medium- and long-term trends and insufficient precise interventions in its prevention and control in order to provide support for formulating targeted prevention and control strategies.

The GBD database was used to extract the incidence, mortality and disability-adjusted life years (DALYs) of pressure ulcers in the general population and in the elderly aged 65 years and above in China from 1990 to 2021. Meanwhile, the age-standardized incidence rate (ASIR), mortality rate (ASMR), and disability-adjusted life rate (ASDR) of the general population were also extracted, and the age-standardized rate (ASRs) corresponding to the elderly population aged 65 years and above were estimated based on the GBD 2021 standard population. The average annual percentage changes (EAPCs) of ASRs were calculated to assess the trends. Bayesian age-period-cohort (BAPC) model was adopted to predict the ASRs and their changing trends of the general and elderly population from 2022 to 2035, respectively.

From 1990 to 2021, the number of pressure ulcer cases in the general population of China increased from 163 508.67 (95%CI: 142 891.48~184 055.10) cases to 397 312.27 (95%CI: 360 058.44~444 875.78) cases, the number of deaths soared from 242.51 (95%CI: 125.26~702.84) cases to 3 131.54 (95%CI: 1 562.60~4 126.85) cases, and DALYs increased from 11 965.10 (95%CI: 7 820.54~21 703.78) to 61 933.91 (95%CI: 36 313.26~77 402.42). During the same period, the EAPCs of ASIR, ASMR and ASDR were 1.06 (95%CI: 0.84~1.28), 3.63 (95%CI: 3.45~3.81), and 2.76 (95%CI: 2.60~2.92), respectively. In 2021, the number of male cases and DALYs were both higher than those of females, while the number of deaths was similar. The ASIR, ASMR and ASDR of the males were all higher than those of the females, and the growth rates of both genders were relatively fast. All indicators were increased significantly with age. It is predicted that by 2035, the ASIR, ASMR, and ASDR in the general Chinese population will continue to rise significantly. From 1990 to 2021, the number of pressure ulcer cases among the elderly (≥65 years old) in China increased from 42 334.95 (95%CI: 35 739.05~48 930.84) to 238 431.65 (95%CI: 212 539.53~264 323.77). The number of deaths rose from 204.97 (95%CI: 93.33~316.60) to 2 871.02 (95%CI: 2 334.39~3 407.66), and DALYs increased from 4 619.74 (95%CI: 2 990.73~6 248.76) to 43 968.28 (95%CI: 37 542.85~50 393.70). The EAPCs of ASIR, ASMR, and ASDR were 2.62 (95%CI: 2.44~2.80), 3.54 (95%CI: 3.38~3.70), and 3.28 (95%CI: 3.14~3.42), all of which increased significantly. In 2021, the number of cases, deaths and DALYs among elderly men and women were similar, but the ASIR, ASMR and ASDR of men were all higher than those of women, and their growth rates were faster. It is projected that by 2035, the ASIR of the elderly population will significantly decline to 146.24(95%CI: 140.35~152.14), with an EAPC of -0.57 (95%CI: -0.64~-0.50), while the ASMR and ASDR will continue to rise and remain higher in males than in females.

From 1990 to 2021, the number of pressure ulcer cases, deaths and DALYs among the general population and the elderly in China increased significantly, and the ASRs rose rapidly. The burden of pressure ulcers increases significantly with age and is higher in men than in women. Predictions show that ASIR in the general population will continue to rise, while that in the elderly will slightly decline. However, the ASMR and ASDR will continue to increase. Countermeasures　Continuously strengthen the prevention and control of pressure ulcers, implement precise intervention for the elderly, especially older men, promote standardized nursing guidelines, enhance training and resource input in primary medical care, and establish a national pressure ulcer monitoring system.

To investigate the ameliorative effect of deferoxamine (DFO) on cognitive impairment in a rat model of type 1 diabetes mellitus (T1DM) and to elucidate the molecular mechanisms underlying cerebral iron overload in T1DM rats.

Thirty-six healthy male SD rats (weighing 180~250 g) were randomly assigned into a blank control group (Ctrl), a T1DM model group and a DFO group, with 12 rats in each group. A single dose of 65 mg/kg streptozotocin (STZ) was intraperitoneally injected to the rats to establish a T1DM model, and those with fasting blood glucose ≥16.7 mmol/L at 3 d later were designated as the T1DM group. Intracerebroventricular administration of DFO (5 μg/kg·d) was given to the DFO group for 28 consecutive days since 21 d after STZ injection. Morris water maze test was carried out to assess the spatial learning and memory abilities. Nissl staining and immunofluorescence assay were applied to observe neuronal morphology and number in the hippocampus and cortex. The iron content in the hippocampus was measured with inductively coupled plasma mass spectrometry (ICP-MS). The expression levels of iron metabolism related proteins were detected with Western blotting.

In the T1DM group, significant declines in learning and memory abilities (P<0.01) and impaired neuronal morphology and reduced neuronal counts in the hippocampal CA1 region (P<0.01), CA3 region (P<0.01), and cortex (P<0.05) were observed when compared with those in the Ctrl group. ICP-MS analysis showed a marked increase in the hippocampal iron content in the T1DM group (P<0.01). Western blot results demonstrated that T1DM rats exhibited obviously up-regulated expression of iron storage proteins FTH and FTL in both the hippocampus (P<0.01, P<0.05) and the cortex (P<0.05, P<0.01), enhanced expression of iron import protein DMT1 in both the hippocampus and the cortex (P<0.05), while decreased expression of iron export protein FPN1 in the hippocampus (P<0.01) and the cortex (P<0.05). DFO treatment significantly ameliorated all above abnormalities.

The declines in learning and memory in T1DM rats are closely associated with neuronal damage induced by cerebral iron overload. Iron import protein DMT1 and export protein FPN1 jointly regulate cerebral iron content in T1DM rats. DFO reduces brain iron levels and mitigates iron overload-mediated neuronal injury by modulating the expression of DMT1 and FPN1.

To investigate the potential therapeutic targets and underlying molecular mechanisms of Z-ligustilide (Z-LIG) in treating retinitis pigmentosa (RP).

By integrating multiple databases, such as GeneCards and TargetNet, common targets for RP and Z-LIG were identified. Protein-protein interaction (PPI) network analysis of the targets was conducted using the STRING database and analyzed in Cytoscape to identify core 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 core targets. The PubChem database was accessed for molecular docking verification to validate the binding affinity between Z-LIG and the core targets. Finally, male rd10 mice were randomly divided into a control group (n=5) and a Z-LIG treatment group (n=5). The protective effect of Z-LIG on the visual function in rd10 mice was verified through visual electrophysiology and behavioral tests. Western blotting and RT-qPCR were utilized to investigate the molecular mechanism of action.

A total of 66 shared targets between RP and Z-LIG were identified through the screening process. PPI network analysis identified 55 key targets. GO enrichment analysis yielded 623 terms, which covering 3 dimensions, including cellular component (CC), molecular function (MF), and biological process (BP), such as inflammatory response. KEGG pathway enrichment analysis further concentrated 124 terms, which were enriched in the PI3K-Akt signaling pathway. Molecular docking suggested that Z-LIG could specifically bind with high affinity to RP-related core targets (such as EGFR and STAT3) via hydrogen bonds. In animal experiments, compared to the control group, the rd10 mice from the Z-LIG group showed a greater preference for the dark environment in the light/dark transition test (P<0.05), exhibited significantly higher amplitudes of A-wave and B-wave in electroretinogram (ERG) (P<0.05), greater number of outer nuclear layers, with fewer apoptotic cells and less microglial activation (P<0.05), demonstrated obviously reduced protein levels of p-PI3K/PI3K and p-AKT/AKT in the retina (P<0.05), and had notably down-regulated mRNA levels of pro-inflammatory cytokines IL-1β and IL-6 (P<0.05).

Z-LIG may exert a protective effect on the retina of rd10 mice by inhibiting the activation of the PI3K/AKT-inflammatory axis, thereby delaying retinal degeneration.