This study explores the role and underlying molecular mechanisms of fucoidan sulfate(FPS) in regulating heme oxygenase-1(Hmox1)-mediated ferroptosis to ameliorate myocardial injury in diabetic cardiomyopathy(DCM) through in vivo and in vitro experiments and network pharmacology analysis. In vivo, a DCM rat model was established using a combination of "high-fat diet feeding + two low-dose streptozotocin(STZ) intraperitoneal injections". The rats were randomly divided into four groups: normal, model, FPS, and dapagliflozin(Dapa) groups. In vitro, a cellular model was created by inducing rat cardiomyocytes(H9c2 cells) with high glucose(HG), using zinc protoporphyrin(ZnPP), an Hmox1 inhibitor, as the positive control. An automatic biochemical analyzer was used to measure blood glucose(BG), serum aspartate aminotransferase(AST), serum lactate dehydrogenase(LDH), and serum creatine kinase-MB(CK-MB) levels. Echocardiography was used to assess rat cardiac function, including ejection fraction(EF) and fractional shortening(FS). Pathological staining was performed to observe myocardial morphology and fibrotic characteristics. DCFH-DA fluorescence probe was used to detect reactive oxygen species(ROS) levels in myocardial tissue. Specific assay kits were used to measure serum brain natriuretic peptide(BNP), myocardial Fe~(2+), and malondialdehyde(MDA) levels. Western blot(WB) was used to detect the expression levels of myosin heavy chain 7B(MYH7B), natriuretic peptide A(NPPA), collagens type Ⅰ(Col-Ⅰ), α-smooth muscle actin(α-SMA), ferritin heavy chain 1(FTH1), solute carrier family 7 member 11(SLC7A11), glutathione peroxidase 4(GPX4), 4-hydroxy-2-nonenal(4-HNE), and Hmox1. Immunohistochemistry(IHC) was used to examine Hmox1 protein expression patterns. FerroOrange and Highly Sensitive DCFH-DA fluorescence probes were used to detect intracellular Fe~(2+) and ROS levels. Transmission electron microscopy was used to observe changes in mitochondrial morphology. In network pharmacology, FPS targets were identified through the PubChem database and PharmMapper platform. DCM-related targets were integrated from OMIM, GeneCards, and DisGeNET databases, while ferroptosis-related targets were obtained from the FerrDb database. A protein-protein interaction(PPI) network was constructed for the intersection of these targets using STRING 11.0, and core targets were screened with Cytoscape 3.9.0. Molecular docking analysis was conducted using AutoDock and PyMOL 2.5. In vivo results showed that FPS significantly reduced AST, LDH, CK-MB, and BNP levels in DCM model rats, improved cardiac function, decreased the expression of myocardial injury proteins(MYH7B, NPPA, Col-Ⅰ, and α-SMA), alleviated myocardial hypertrophy and fibrosis, and reduced Fe~(2+), ROS, and MDA levels in myocardial tissue. Furthermore, FPS regulated the expression of ferroptosis-related markers(Hmox1, FTH1, SLC7A11, GPX4, and 4-HNE) to varying degrees. Network pharmacology results revealed 313 potential targets for FPS, 1 125 targets for DCM, and 14 common targets among FPS, DCM, and FerrDb. Hmox1 was identified as a key target, with FPS showing high docking activity with Hmox1. In vitro results demonstrated that FPS restored the expression levels of ferroptosis-related proteins, reduced intracellular Fe~(2+) and ROS levels, and alleviated mitochondrial structural damage in cardiomyocytes. In conclusion, FPS improves myocardial injury in DCM, with its underlying mechanism potentially involving the regulation of Hmox1 to inhibit ferroptosis. This study provides pharmacological evidence supporting the therapeutic potential of FPS for DCM-induced myocardial injury.
Animals ; Ferroptosis/drug effects* ; Rats ; Diabetic Cardiomyopathies/physiopathology* ; Male ; Rats, Sprague-Dawley ; Polysaccharides/pharmacology* ; Heme Oxygenase-1/genetics* ; Myocytes, Cardiac/metabolism* ; Myocardium/pathology* ; Humans ; Cell Line ; Heme Oxygenase (Decyclizing)

Objective:To investigate the effect of capsaicin on lipopolysaccharide(LPS)-induced microglial inflammatory response by modulating silent mating type information regulation 2 homolog 1(SIRT1)/high mobility group box-1 protein(HMGB1)/nuclear factor κB(NF-κB)signaling pathway.Methods:BV2 mouse microglia were cultured in vitro,and pretreated with capsaicin(10 μmol/L),SIRT1 inhibitor EX527(100 μmol/L),capsaicin+EX527(10 μmol/L capsaicin+100 μmol/L EX527)for 3 hours,and then treated with LPS(1 μg/ml)for 24 hours,the corresponding groups were LPS group,LPS+capsaicin group,LPS+EX527 group,and LPS+capsaicin+EX527 group,a normal cultured control group(Control)was also set up.Morphological changes of BV2 cells in each group were observed under a microscope;CCK-8 method was used to detect the viability of BV2 cells in each group;immunofluo-rescence staining was applied to detect the positive expressions of ionic calcium adaptor protein(Iba-1)and the polarization of M1(positive for CD16/32)/M2(positive for CD206)subtypes in BV2 cells in each group;ELISA was applied to detect levels of inflamma-tory factors in BV2 cells in each group;Western blot and co-immunoprecipitation were applied to detect expression of SIRT1/HMGB1/NF-κB signaling pathway-related proteins in each group.Results:Compared with Control group,BV2 cells in LPS group were atro-phied,with shortened processes,the cell viability,and levels of IL-1β,TNF-α and IL-6 were increased,Iba-1 positive expression of BV2 cells and M1 type BV2 cells were increased,while expression of SIRT1 protein was decreased,expressions of acetylated(ace)-HMGB1 protein,cytoplasmic HMGB1 protein and nuclear NF-κB protein were increased(P<0.05);after capsaicin pretreatment,the above conditions were improved,and the M2 type BV2 cells were increased;adding EX527 pretreatment on the basis of capsaicin pre-treatment,the above conditions were all aggravated.Conclusion:Capsaicin can inhibit the inflammatory response induced by LPS-in-duced BV2 cell activation,which may be achieved by modulating the SIRT1/HMGB1/NF-κB signaling pathway.

Objective:To investigate the impact of age and gender on kinetic visual acuity (KVA) and static visual acuity (SVA) in school children accepting physical activities. Methods:From May, 2018 to September, 2019, 1465 school children from various schools of Suzhou City were measured SVA and KVA with standard logarithmic visual chart and KVA detector. Results:KVA increased with age as six to nine years old, and decreased as eleven to 14. KVA was better in boys than in girls (t = 5.147, P < 0.001). SVA decreased with age as six to eight years old, fluctuated as eight to eleven, and decreased as eleven to 14, especially for girls. SVA was also better in boys than in girls (t = 3.692, P < 0.001). The prevalence of myopia increased with age as six to eight years old, and increased more significantly as eleven to 14. There was a low positive correlation between age and KVA (r = 0.046, P < 0.05), and a moderate negative correlation between age and SVA (r = -0.364, P < 0.05). KVA was positively correlated with SVA (r = 0.409, P < 0.05). Age, gender, KVA, right SVA and left SVA were the factors related with SVA. Conclusion:Vision is better in school boys than in girls. There is positive correlation between age and KVA. It is necessary to develop physical exercise according to age and gender to improve vision development, especially at the sensitive stage of visual function.

Objective To compare the effectiveness and safety of ultramini percutaneous nephrolithotomy (UMP) and retrograde intrarenal surgery (RIRS) in treatment of moderate-sized (about 1-2 cm) renal lower caliceal calculi.Methods From March 2015 to December 2016,patients in our hospital scheduled for surgery due to renal lower caliceal calculi with the greatest diameter of 10-22 mm were prospectively analyzed.Patients were randomized into two groups according to the random number table.Group UMP's operational channel was only F14 and the nephroscope's diameter was 1 mm.200 μm holmium laser lithotripsy was used to break the stones which was rushed out by eddy cuurent.In Group RIRS,all patients needed placing a F6 double J stent preoperatively for two weeks.A flexible ureteroscope sheath required imbedding intraoperatively.The stones were smashed by 200 μm holmium laser lithotripsy through the WOLF flexible ureteroscope.The intraoperative and postoperative datas including stone-free status and the complications were compared.Results 100 patients were enrolled in the study 50 patients in Group UMP,28 were male and 22 were female,mean age was 43.4 ± 7.9 years old.Mean stone size was 14.5 ±3.0 mm(range 10-22 mm).Among them,18 cases were complicated with mild and moderate hydronephrosis.The other 50 cases were allocated to Group RIRS,including 31 males and 19 females.Their mean age was 44.5 ± 8.3 years old and mean stone size was 13.7 ± 3.1 mm (range 10-21 mm).Among them,16 cases were complicated with mild and moderate hydronephrosis.No statistically significant difference were seen between the two groups (P ＞ 0.05).After three months' follow-up,one-time stone free rate(SFR) of UMP group was 94.0％ (47/50),which was significantly more superior than the 72.0％ (36/50) of the RIRS group(P ＜ 0.05).The intraoperative decrease in hemoglobin were (7.8 ± 3.3) g/L vs.(3.1 ± 3.4) g/L,and operating time(26.5 ± 6.1) min vs.(43.3 ± 6.3) min.Significant differences were also seen between the two groups(P ＜0.05).There was more blood loss and less operating time in the group of UMP.The hospital stay,delayed hemorrhage and postoperative fever between the UMP and RIRS groups were (4.3±1.3)d vs.(3.24 ± 1.21)d,8.0％ (4/50)vs.0(0/50),16.0％ (8/50)vs.12.0％ (6/50) respectively.No significant differences were seen (P ＞ 0.05).Conclusions Both UMP and RIRS procedures are effective and safe in the treatment of moderate-sized renal lower caliceal calculi.Compared with RIRS,UMP may be more effective and has less operating time,however wtih more intraoperative blood loss.