Objective:To explore the association between spicy food intake and the risk for cardio/cerebrovascular diseases.Methods:Data were collected from the China Kadoorie Biobank project conducted in Pengzhou, Sichuan Province. Using the Cox proportional hazards regression model, we analyzed the associations of the frequency of spicy food intake, spicy level, types of spicy food, and the age when regular intake of spicy food began (intake in 1 day/week), with the risk for cardio/cerebrovascular disease. Furthermore, the associations with the risks for ischemic heart disease (IHD) and cerebrovascular diseases, as well as the risk of ischemic stroke (IS) and hemorrhagic stroke (HS) were analyzed.Results:A total of 54 859 study participants were included in the study, in whom 49 320 had spicy food intake (89.90%). In these participants, 37 680 (68.69%) had spicy food intake in 6-7 days/week, 5 036 (9.18%) had spicy food intake in 1-5 days/week, and 6 604 (12.03%) had spicy food intake once a week; 5 539 (10.10%) had never/almost never had spicy food intake. After adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of spicy food was associated with reduced risks for IHD (intake in 6-7 days/week: HR=0.86, 95% CI: 0.78-0.95), cerebrovascular diseases (intake in 6-7 days/week: HR=0.88, 95% CI: 0.81-0.96), and IS (intak in 6-7 days/week: HR=0.85, 95% CI: 0.76-0.95). With the increase of spicy food intake frequency, the risk for cardio/cerebrovascular disease decreased (intake in 1-5 days/week: HR=0.91, 95% CI: 0.85-0.98; intake in 6-7 days/week: HR=0.89, 95% CI: 0.84-0.94) (trend test P<0.001). However, no statistical association was found between spicy food intake and the risk for HS. In terms of spicy level, after adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of spicy food was associated with reduced risk for cardio/cerebrovascular disease (moderate: HR=0.86, 95% CI: 0.82-0.90) and cerebrovascular disease (moderate: HR=0.90, 95% CI: 0.84-0.97). With the increase of spicy level, the risk for IHD decreased (moderate: HR=0.86, 95% CI: 0.79-0.93; strong: HR=0.84, 95% CI: 0.74-0.95) (trend test P<0.001). After adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of any type of spicy food was associated with reduced risk for cardio/cerebrovascular disease, IHD, and cerebrovascular disease. Regulat intake of spicy food from age 0-10 years was associated with reduced risk for cardio/cerebrovascular disease, IHD, and cerebrovascular disease. Regular intake of spicy food from age 11-20 years reduced the risk for cardio/cerebrovascular disease and IHD. There was no significant association between the regular intake of spicy food from age 21-79 years and the risks for cardio/cerebrovascular disease, IHD and cerebrovascular disease. Conclusion:The intake of spicy food could reduced the risk for cardio/cerebrovascular diseases, IHD, cerebrovascular diseases and IS in residents aged 30-79 years in Sichuan.

Objective To identify potential biomarkers for proliferative diabetic retinopathy(PDR)using proteomics and transcriptomics data.Methods In this study,the proteomics dataset(PXD046630)and two transcriptomics datasets(GSE60436 and GSE102485)were derived from the aqueous humor samples and fibrovascular membranes of PDR patients,respectively.Differentially expressed genes(DEGs)were identified via R software,specifically the limma and edgeR pack-ages.The shared DEGs between PXD046630 and GSE60436 were analyzed via protein-protein interaction(PPI),Gene On-tology(GO)enrichment,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.The key DEGs were validated in GSE102485 via receiver operating characteristic(ROC)curve analysis.A quantitative polymerase chain reaction(qPCR)assay was used to confirm the mRNA of these candidate biomarkers in human retinal microvascular endothelial cells(HRMECs)cultured in high glucose and low oxygen conditions.Results A total of 59 shared DEGs and 26 hub genes were identified from the PXD046630 and GSE60436 datasets.KEGG analysis revealed that six pathways,inclu-ding extracellular matrix-receptor interaction,proteoglycans in cancer,and complement and coagulation cascades,were enriched in 12 key DEGs.Fibronectin 1(FN1),tissue inhibitor of metalloproteinase 3(TIMP3),complement factor H(CFH),decorin(DCN),and lipoprotein receptor-related protein-2(LRP2)were identified as potential biomarkers on the basis of their AUC values being greater than 0.900(CI≥95％).The mRNA expression levels of FN1,CFH,and LRP2 were significantly increased in HRMECs cultured in high glucose and low oxygen conditions.Conclusion FN1,CFH,and LRP2 are potential biomarkers for PDR,and further studies are needed to explore their roles and therapeutic potential in PDR.

Idiopathic pulmonary fibrosis (IPF) is a progressive disease lacking effective therapy. Metformin, an antidiabetic medication, has shown promising therapeutic properties in preclinical fibrosis models; however, its precise cellular targets and associated mechanisms in fibrosis resolution remain incompletely defined. Most research on metformin's effects has focused on mesenchymal and inflammatory responses with limited attention to epithelial cells. In this study, we utilized Sftpc lineage-traced and Fgfr2b conditional knockout mice, along with BMP2/PPARγ and AMPK inhibitors, to explore metformin's impact on alveolar epithelial cells in a bleomycin-induced pulmonary fibrosis model and cell culture. We found that metformin increased the proliferation and differentiation of alveolar type 2 (AT2) cells, particularly the recently identified injury-activated alveolar progenitors (IAAPs)-a subpopulation characterized by low SFTPC expression but enriched for PD-L1. Single-cell RNA sequencing revealed a reduction in apoptosis among mature AT2 cells. Interestingly, metformin's therapeutic effects were not significantly affected by BMP2 or PPARγ inhibition, which blocked the lipogenic differentiation of myofibroblasts. However, Fgfr2b deletion in Sftpc lineage cells significantly impaired metformin's ability to promote fibrosis resolution, a process linked to AMPK signaling. In conclusion, metformin alleviates fibrosis by directly activating AT2 cells, especially the IAAPs, through a mechanism that involves AMPK and FGFR2b signaling, but is largely independent of BMP2/PPARγ pathways.

Objective To comprehensively understand the map of transcripts during mitosis and their regulatory mechanisms of HAP 1 cells by conducting transcriptome sequencing analysis after being released by mitotic synchro-nization arrest.Methods HAP1 cells were subjected to mitotic synchronous arrest with nocodazole and samples were collected after 0,20,80 min release,and RNA sequencing(RNA-seq)were performed.The transcriptome data was cleaned and the differentially expressed genes,expression trend clustering and functional enrichment com-bined with the protein interaction network were analyzed to explore the changes of signaling pathways in HAP 1 cells during mitosis.Results The transcriptome of HAP1 cells after synchronous release from mitosis underwent significant changes in time series,and differential gene cluster analysis revealed four gene clusters were enriched in important biological processes such as p53 signaling and cytoplasmic translation.Conclusions The transcriptome time-dependent dynamic changes during mitosis in HAP1 cells are coordinated regulation of key signaling pathways including cellular stress response,translational control and chromatin remodeling,ensuring a balance between growth and stress response upon mitotic exit.

Objective:To explore the effects of fluoride exposure from early life to adulthood on learning and memory abilities of offspring mice and the expression of genes related to the NOTCH signaling pathway in hippocampal tissue.Methods:Thirty 8-week-old (body weight 18 - 24 g) clean-grade C57BL/6J mice, half male and half female, were randomly divided into 3 groups by body weight and then paired 1 ∶ 1 (10 mice per group). Successfully pregnant female mice were given tap water containing 0 (control), 20, or 40 mg/L sodium fluoride (NaF) during gestation and lactation until weaning. Offspring mice were exposed to fluoride until 12 weeks of age after weaning at 21 days, with fluoride exposure doses identical to those of the maternal mice. The 24-hour urine of the offspring mice was collected to determine urine fluoride level using fluoride ion selective electrode method, and Morris water maze was used to evaluate their spatial learning and memory ability. The offspring mice were euthanized under anesthesia, and protein and RNA were extracted from hippocampal tissue. Western blot was used to detect the protein expression levels of postsynaptic density protein 95 (PSD95) and synapsin-1 (SYN-1) in hippocampal tissues. Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expression levels of NOTCH signaling pathway related genes (including NOTCH1, DLL3, HES5) in hippocampal tissue.Results:Compared with the control group [(0.86 ± 0.25) mg/L], the offspring mice of the 20 and 40 mg/L NaF groups had higher urinary fluoride levels [(3.77 ± 0.51), (6.04 ± 1.63) mg/L, P < 0.05]. The Morris water maze results showed that compared with the control group, the offspring mice in the 20 and 40 mg/L NaF groups had longer escape latency (day 5), fewer platform crossings, and shorter target platform dwell time ( P < 0.05). Compared with the control group, the protein expression level of synaptic related protein PSD95 in the hippocampal tissues of the offspring mice in the 40 mg/L NaF group was lower, and the protein expression level of SYN-1 in the 20 and 40 mg/L NaF groups was lower ( P < 0.05). The protein expression level of NOTCH1, a gene related to the NOTCH signaling pathway, was lower in the hippocampal tissue of offspring mice in the 40 mg/L NaF group. The mRNA and protein expression levels of DLL3 and HES5 were also lower in the 20 and 40 mg/L NaF groups, and the mRNA expression level of HES5 in the 40 mg/L NaF group was lower than that in the 20 mg/L NaF group ( P < 0.05). Conclusions:Exposure to fluoride from early life to adulthood can lead to a decline in the learning and memory abilities of offspring mice, as well as a decrease in the expression of synaptic related proteins. The mechanism of action may be related to the inhibition of the NOTCH signaling pathway in hippocampal tissue.

BACKGROUND: Doxorubicin hydrochloride (DOX) is extensively used in the treatment of various tumors. However, its clinical application is limited due to dose-dependent cardiotoxicity. Currently, few effective strategies exist to mitigate or eliminate DOX-induced cardiomyopathy (DIC). Although ferroptosis is implicated in DIC and its inhibition partially alleviates the condition, the direct targets of DOX in the progression of cardiotoxicity remain unclear. This study aimed to discover the direct targets of DOX in ferroptosis-mediated DIC. METHODS: A DOX pulldown assay was performed to identify proteins specifically binding to DOX in murine hearts, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify candidate proteins. A cardiac injury mouse model was established by DOX treatment. Based on this, multiple ferroptosis biomarkers were detected by flow cytometry, quantitative real-time polymerase chain reaction, western blotting, immunochemistry, etc. Besides, specific activator and inhibitor of signaling pathways were applied to illuminate molecular mechanisms. RESULTS: Glutathione S-transferase P1 (GSTP1) was identified as a DOX target. GSTP1 activity was inhibited in DOX-treated cardiomyocytes, while its overexpression significantly alleviated DIC. Moreover, GSTP1 overexpression inhibited acyl-CoA synthetase long-chain family member 4 (ACSL4)-dependent ferroptosis. Mechanistically, GSTP1 overexpression suppressed c-Jun N-terminal kinase (JNK) phosphorylation, thereby reducing reactive oxygen species (ROS) production and inhibiting ferroptosis in DIC. CONCLUSIONS This study identifies the DOX/GSTP1/JNK axis as a critical pathway mediating ACSL4-dependent ferroptosis in DIC. GSTP1 is highlighted as a potential key mediator of ferroptosis and a promising therapeutic target for DIC.

Objective:To study on the relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus.Methods:120 elderly patients with GDM were treated in Obstetrics Department of Yantai Yuhuangding Hospital from Jun. 2019 to Jun. 2021. The healthy women (58 cases) and normal pregnant women (58 cases) were selected as the control group at the same time, and the clinical data and serum samples of subjects were collected. The levels of serum SC, SV, FPG and HbAlc were measured, the HOMA-IR of the aged GDM was calculated, and the difference of serum index expression among the subjects in each group was analyzed. The relationship between serum SC, SV and insulin resistance was analyzed by Person correlation, and the risk factors of GDM were analyzed by Logistic regression.Results:: There was significant difference in HbAlc, HOMA-IR, FPG, SC, and SV among the three groups. The levels of each index in the elderly GDM group were significantly higher than those in the normal pregnancy group and the healthygroup ( P<0.05). Pearson analysis showed that SC ( P=0.558, 0.626, 0.672), SV ( P=0.576, 0.663, 0.696) was positively correlated with HbAlc, HOMA-IR, FPG in elderly patients with GDM ( P<0.001). Logistic regression analysis showed that SC, SV was risk factors for GDM in elderly pregnant women. Conclusion:There is a positive correlation between serum SC, SV level and insulin resistance, which are risk factors for GDM in elderly pregnant women.

Objective:To explore the effects of fluoride exposure from early life to adulthood on learning and memory abilities of offspring mice and the expression of genes related to the NOTCH signaling pathway in hippocampal tissue.Methods:Thirty 8-week-old (body weight 18 - 24 g) clean-grade C57BL/6J mice, half male and half female, were randomly divided into 3 groups by body weight and then paired 1 ∶ 1 (10 mice per group). Successfully pregnant female mice were given tap water containing 0 (control), 20, or 40 mg/L sodium fluoride (NaF) during gestation and lactation until weaning. Offspring mice were exposed to fluoride until 12 weeks of age after weaning at 21 days, with fluoride exposure doses identical to those of the maternal mice. The 24-hour urine of the offspring mice was collected to determine urine fluoride level using fluoride ion selective electrode method, and Morris water maze was used to evaluate their spatial learning and memory ability. The offspring mice were euthanized under anesthesia, and protein and RNA were extracted from hippocampal tissue. Western blot was used to detect the protein expression levels of postsynaptic density protein 95 (PSD95) and synapsin-1 (SYN-1) in hippocampal tissues. Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expression levels of NOTCH signaling pathway related genes (including NOTCH1, DLL3, HES5) in hippocampal tissue.Results:Compared with the control group [(0.86 ± 0.25) mg/L], the offspring mice of the 20 and 40 mg/L NaF groups had higher urinary fluoride levels [(3.77 ± 0.51), (6.04 ± 1.63) mg/L, P < 0.05]. The Morris water maze results showed that compared with the control group, the offspring mice in the 20 and 40 mg/L NaF groups had longer escape latency (day 5), fewer platform crossings, and shorter target platform dwell time ( P < 0.05). Compared with the control group, the protein expression level of synaptic related protein PSD95 in the hippocampal tissues of the offspring mice in the 40 mg/L NaF group was lower, and the protein expression level of SYN-1 in the 20 and 40 mg/L NaF groups was lower ( P < 0.05). The protein expression level of NOTCH1, a gene related to the NOTCH signaling pathway, was lower in the hippocampal tissue of offspring mice in the 40 mg/L NaF group. The mRNA and protein expression levels of DLL3 and HES5 were also lower in the 20 and 40 mg/L NaF groups, and the mRNA expression level of HES5 in the 40 mg/L NaF group was lower than that in the 20 mg/L NaF group ( P < 0.05). Conclusions:Exposure to fluoride from early life to adulthood can lead to a decline in the learning and memory abilities of offspring mice, as well as a decrease in the expression of synaptic related proteins. The mechanism of action may be related to the inhibition of the NOTCH signaling pathway in hippocampal tissue.

Objective:To explore the association between spicy food intake and the risk for cardio/cerebrovascular diseases.Methods:Data were collected from the China Kadoorie Biobank project conducted in Pengzhou, Sichuan Province. Using the Cox proportional hazards regression model, we analyzed the associations of the frequency of spicy food intake, spicy level, types of spicy food, and the age when regular intake of spicy food began (intake in 1 day/week), with the risk for cardio/cerebrovascular disease. Furthermore, the associations with the risks for ischemic heart disease (IHD) and cerebrovascular diseases, as well as the risk of ischemic stroke (IS) and hemorrhagic stroke (HS) were analyzed.Results:A total of 54 859 study participants were included in the study, in whom 49 320 had spicy food intake (89.90%). In these participants, 37 680 (68.69%) had spicy food intake in 6-7 days/week, 5 036 (9.18%) had spicy food intake in 1-5 days/week, and 6 604 (12.03%) had spicy food intake once a week; 5 539 (10.10%) had never/almost never had spicy food intake. After adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of spicy food was associated with reduced risks for IHD (intake in 6-7 days/week: HR=0.86, 95% CI: 0.78-0.95), cerebrovascular diseases (intake in 6-7 days/week: HR=0.88, 95% CI: 0.81-0.96), and IS (intak in 6-7 days/week: HR=0.85, 95% CI: 0.76-0.95). With the increase of spicy food intake frequency, the risk for cardio/cerebrovascular disease decreased (intake in 1-5 days/week: HR=0.91, 95% CI: 0.85-0.98; intake in 6-7 days/week: HR=0.89, 95% CI: 0.84-0.94) (trend test P<0.001). However, no statistical association was found between spicy food intake and the risk for HS. In terms of spicy level, after adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of spicy food was associated with reduced risk for cardio/cerebrovascular disease (moderate: HR=0.86, 95% CI: 0.82-0.90) and cerebrovascular disease (moderate: HR=0.90, 95% CI: 0.84-0.97). With the increase of spicy level, the risk for IHD decreased (moderate: HR=0.86, 95% CI: 0.79-0.93; strong: HR=0.84, 95% CI: 0.74-0.95) (trend test P<0.001). After adjusting for multiple confounding factors, compared with those who never/almost never had spicy food intake, intake of any type of spicy food was associated with reduced risk for cardio/cerebrovascular disease, IHD, and cerebrovascular disease. Regulat intake of spicy food from age 0-10 years was associated with reduced risk for cardio/cerebrovascular disease, IHD, and cerebrovascular disease. Regular intake of spicy food from age 11-20 years reduced the risk for cardio/cerebrovascular disease and IHD. There was no significant association between the regular intake of spicy food from age 21-79 years and the risks for cardio/cerebrovascular disease, IHD and cerebrovascular disease. Conclusion:The intake of spicy food could reduced the risk for cardio/cerebrovascular diseases, IHD, cerebrovascular diseases and IS in residents aged 30-79 years in Sichuan.

Objective:To study on the relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus.Methods:120 elderly patients with GDM were treated in Obstetrics Department of Yantai Yuhuangding Hospital from Jun. 2019 to Jun. 2021. The healthy women (58 cases) and normal pregnant women (58 cases) were selected as the control group at the same time, and the clinical data and serum samples of subjects were collected. The levels of serum SC, SV, FPG and HbAlc were measured, the HOMA-IR of the aged GDM was calculated, and the difference of serum index expression among the subjects in each group was analyzed. The relationship between serum SC, SV and insulin resistance was analyzed by Person correlation, and the risk factors of GDM were analyzed by Logistic regression.Results:: There was significant difference in HbAlc, HOMA-IR, FPG, SC, and SV among the three groups. The levels of each index in the elderly GDM group were significantly higher than those in the normal pregnancy group and the healthygroup ( P<0.05). Pearson analysis showed that SC ( P=0.558, 0.626, 0.672), SV ( P=0.576, 0.663, 0.696) was positively correlated with HbAlc, HOMA-IR, FPG in elderly patients with GDM ( P<0.001). Logistic regression analysis showed that SC, SV was risk factors for GDM in elderly pregnant women. Conclusion:There is a positive correlation between serum SC, SV level and insulin resistance, which are risk factors for GDM in elderly pregnant women.