[Effect of Xiongcan Yishen Formula on ferroptosis in mouse TM3 Leydig cells after oxidative stress injury].

OBJECTIVE: To investigate the effects of Xiongcan Yishen Formula (XYF) on ferroptosis in mouse TM3 Leydig cells after oxidative stress injury (OSI) induced by H2O2.　Methods: An oxidative stress injury model was established in mouse TM3 Leydig cells using H2O2 induction. The modeled TM3 cells were randomly divided into OSI group, XYF group, the ferroptosis inhibitor Ferrostatin-1 (F-1) group, and F-1+XYF group, which were respectively intervened with blank serum, 20% drug-containing serum, 2µmol/L F-1, and 2µmol/L F-1+ 20% drug-containing serum. A control group (normal TM3 cells + blank serum) was also set up. The morphology of cells in each group was observed, and the levels of testosterone, superoxide dismutase (SOD), reactive oxygen species (ROS), malondialdehyde (MDA), ferritin heavy chain 1 (FTH1), solute carrier family 7 member 11 (SLC7A11), glutathione (GSH), glutathione peroxidase 4 (GPX4), fatty acid CoA ligase 4 (FACL4), total iron ions, and ferrous ions were detected. RESULTS: Compared with the model group, the control group showed significantly decreased expression of ROS, MDA, FACL4, total iron, and ferrous ions (P<0.05), and significantly increased levels of testosterone, SOD, GSH, FTH1, SLC7A11, and GPX4 (P<0.05). The male silkworm kidney-tonifying formula group significantly promoted testosterone secretion by TM3 cells and upregulated the expression of FTH1, SLC7A11, GPX4, GSH, and SOD in TM3 cells (P<0.05), while significantly downregulating ROS, MDA, FACL4, total iron ions, and ferrous ions (P<0.05). CONCLUSION: Following H2O2 exposure, oxidative stress can induce ferroptosis in mouse TM3 Leydig cells. XYF can antagonize OSI and ferroptosis in TM3 cells by activating the SLC7A11/GSH/GPX4 axis, which may underlie the mechanism of XYF in the treatment of male late-onset hypogonadism.

Emerging strategies for treating autoimmune disease with genetically modified dendritic cells.

Gene editing of living cells has become a crucial tool in medical research, enabling scientists to address fundamental biological questions and develop novel strategies for disease treatment. This technology has particularly revolutionized adoptive transfer cell therapy products, leading to significant advancements in tumor treatment and offering promising outcomes in managing transplant rejection, autoimmune disorders, and inflammatory diseases. While recent clinical trials have demonstrated the safety of tolerogenic dendritic cell (TolDC) immunotherapy, concerns remain regarding its effectiveness. This review aims to discuss the application of gene editing techniques to enhance the tolerance function of dendritic cells (DCs), with a particular focus on preclinical strategies that are currently being investigated to optimize the tolerogenic phenotype and function of DCs. We explore potential approaches for in vitro generation of TolDCs and provide an overview of emerging strategies for modifying DCs. Additionally, we highlight the primary challenges hindering the clinical adoption of TolDC therapeutics and propose future research directions in this field.

Circulating serum fibroblast growth factor 21 as risk and prognostic biomarker of retinal artery occlusion.

To evaluate the predictive and prognostic value of fibroblast growth factor 21 (FGF21) levels in retinal artery occlusion (RAO) patients. In this case-control study, serum FGF21 levels were detected by using the ELISA method. Multivariable logistic regression analyses were performed to evaluate the significance of FGF21 in assessing the risk of developing RAO and its impact on vision and concurrent ischemic stroke. Compared with control group, serum FGF21 levels were significantly higher (median [IQR] = 230.90[167.40,332.20] pg/ml) in RAO patients. Multivariate logistic regression analysis showed that elevated serum FGF21 levels were associated with a higher risk of RAO occurrence (P = 0.025, OR [95%CI] = 9.672 [2.573, 36.359]) after adjustment for multiple confounding factors. Higher serum FGF21 levels were negatively associated with visual acuity improvement (P = 0.029, OR [95%CI] = 0.466[0.235, 0.925]) and positively correlated with concurrent ischemic stroke (P = 0.04, OR [95% CI] = 1.944[1.029, 3.672]) in RAO patients. Elevated serum FGF21 levels could promote the development of RAO and indicate worse visual prognosis and increase the risk of concurrent ischemic stroke, which might help clinicians early diagnose and treat RAO patients.

[Xiongcan Yishen Formula up-regulates the clock gene expressions in the testis tissue of the rats with late-onset hypogonadism].

OBJECTIVE: To investigate the effect of Xiongcan Yishen Formula (XYF) on the expressions of the clock genes in the testis tissue of the rats with late-onset hypogonadism (LOH). METHODS: Forty-eight 8-week-old male SD rats were randomly divided into 6 groups, normal control, model control, testosterone propionate (TP), and low-, medium- and high-dose XYF. The LOH model was made in the later 5 groups of rats by intraperitoneal injection of D-galactose at 480 mg/kg/d for 56 successive days, while the normal controls were injected with the same volume of normal saline. After modeling, the rats in the low-, medium- and high-dose XYF groups were treated intragastrically with XYF at 10.4, 20.8 and 41.6 g/kg/d, bid, respectively, those in the normal and model control groups with the same volume of distilled water, and those in the TP group intramuscularly with TP at 5.21 mg/kg/d, qd alt, all for 28 days. After treatment, the supernatant was obtained for measurement of the serum T level by ELISA, and the testis tissue collected for determination of the mRNA and protein expressions of BMAL1, NR1D1, PER2, CRY1, StAR and CYP11A1 by RT-qPCR and Western blot. RESULTS: Compared with the normal controls, the rats in the LOH model control group showed significantly decreased serum T and mRNA and protein expressions of BMAL1, NR1D1, PER2, CRY1, StAR and CYP11A1 (P < 0.05). In comparison with the findings in the model controls, the T level was remarkably increased in the TP and XYF groups (P < 0.05), the expressions of StAR mRNA and CYP11A1 mRNA and protein markedly up-regulated in the high-dose XYF group (P < 0.05), and so was the expression of the StAR protein in the XYF and TP groups (P < 0.05), those of BMAL1 and NR1D1 proteins and PER2 mRNA and protein in the high-dose XYF group (P < 0.05), those of BMAL1 mRNA and CRY1 protein in the medium- and high-dose XYF groups (P < 0.05), that of NR1D1 mRNA in the XYF and TP groups (P < 0.05), and that of CRY1 mRNA in the medium- and high-dose XYF and TP groups (P < 0.05). CONCLUSION: Xiongcan Yishen Formula could up-regulate the expressions of the clock genes in the testis tissue of the LOH rats and increase the serum T level as well, which may underlie the mechanisms of Xiongcan Yishen Formula acting on LOH.

[Potential mechanisms of Jiarong Tablets in the treatment of late-onset hypogonadism in men: An exploration based on network pharmacology].

OBJECTIVE: To study the therapeutic targets and related signaling pathways of Jiarong Tablets (JRT) in the treatment of late-onset hypogonadism (LOH) in males by network pharmacology, and further analyze its potential action mechanism. METHODS: Using the Chinese Medicine System Pharmacology Analysis Platform (TCMSP), we obtained the active ingredients and therapeutic targets of JRT, disease targets of LOH through the GeneCards and OMIM databases, and drug-disease common targets, followed by drawing a Vennny's diagram of the common targets. We constructed a protein-protein interaction (PPI) network of the common targets using STRING and an intersection network of JRT active ingredients-LOH-targets with Cytoscape 3.7.2, performed GO bio-functional and KEGG enrichment analyses of the common targets using the R-Language software, and identified the potential signaling pathways of JRT acting on LOH. RESULTS: Totally, we obtained 80 bioactive ingredients from JRT and 64 common targets of LOH, with IL-6, INS, AKT1, JUN and MAPK8 as the core targets in the order of the frequency occurrences. GO and KEGG analyses showed that these targets mainly involved the MAPK, HIF-1, Ras and ErbB signaling pathways. CONCLUSION: JRT acts on LOH with multiple targets, through multiple routes and at multiple levels, which is related to the expression of testosterone synthetase, oxidative stress and apoptosis of Leydig cells.

[Establishment and evaluation of a rat model of late-on-set hypogonadism using D-galactose].

OBJECTIVE: To explore the feasibility and duration of establishing a model of late-on-set hypogonadism (LOH) in rats using D-galactose (D-gal) and the optimal concentration of D-gal in modeling. METHODS: Thirty-five 8-week-old male SD rats were randomly divided into seven groups of an equal number to receive intraperitoneal injection of normal saline (normal control) or D-gal at 480 mg/kg/d (high-dose), 240 mg/kg/d (medium-dose) and 120 mg/kg/d (low-dose) for 6 or 8 weeks respectively. Another five 18-month-old SD rats were taken as positive controls. After modeling, the animals were subjected to tail suspension test and mating test, weighed, and then sacrificed for examination of the levels of fasting blood glucose (FBG) and serum testosterone (T), testis index and semen parameters, determination of the expressions of the BMAL1 and NR1D1 proteins in the testis tissue by Western blot, and observation of the pathological and ultrastructural changes in the testis tissue under the light microscope and transmission electron microscope. RESULTS: Compared with the normal controls, the serum T level was significantly decreased in the 6-week medium-dose D-gal (6WMD), 6-week high-dose D-gal (6WHD), 8WMD, 8WHD and positive control groups (P < 0.05), and so were sperm concentration and the percentage of progressively motile sperm (PMS) in the 8WHD and positive control groups (P < 0.05), and the numbers of captures and ejaculations in all the model rats and positive controls (P < 0.05), while the body weight was markedly increased in the 8-week low-dose D-gal (8WLD), 8WMD and 8WHD groups (P < 0.05), and so were the tail suspension time in the 6WHD, 8WHD, 8WMD, 8WLD and positive control groups (P < 0.05) and the FBG level in all the model rats and positive controls (P < 0.05). The expression of the BMAL1 protein was significantly decreased in the 6WHD, 8WLD, 8WMD, 8WHD and positive control groups (P < 0.05), and so was that of NR1D1 in the 8WMD, 8WHD and positive control groups (P < 0.05). Compared with the positive controls, no statistically significant differences were observed in the serum T level in the 6WHD and 8WHD groups (P > 0.05), the testis index in the 8WMD and 8WHD groups (P > 0.05), sperm concentration, percentage of FMS, time of tail suspension and numbers of captures and ejaculations in the 8WHD group (P > 0.05), the level of FBG in the 6WMD, 6WHD, 8WLD, 8WMD and 8WHD groups (P > 0.05), the expression of BMAL1 in the 6WHD, 8WLD, 8WMD and 8WHD groups (P > 0.05), and that of NR1D1 in the 6WLD, 8WMD and 8WHD groups (P > 0.05). Pathological and ultrastructural changes in the testis tissue were observed in all the model rats and positive controls, most significantly in the 8WHD and positive control groups. CONCLUSION: It is feasible to establish an LOH model in male SD rats by intraperitoneal injection of D-gal, most recommendably at the centration of 480 mg/kg/d for 8 weeks.

Preparation of magnetite-loaded silica microspheres for solid-phase extraction of genomic DNA from soy-based foodstuffs.

Solid-phase extraction has been widely employed for the preparation of DNA templates for polymerase chain reaction (PCR)-based analytical methods. Among the variety of adsorbents studied, magnetically responsive silica particles are particularly attractive due to their potential to simplify, expedite, and automate the extraction process. Here we report a facile method for the preparation of such magnetic particles, which entails impregnation of porous silica microspheres with iron salts, followed by calcination and reduction treatments. The samples were characterized using powder X-ray diffractometry (XRD), scanning electron microscopy (SEM), nitrogen adsorption/desorption isotherms, and vibrating sample magnetometry (VSM). XRD data show that magnetite nanocrystals of about 27.2 nm are produced within the pore channels of the silica support after reduction. SEM images show that the as-synthesized particles exhibit spherical shape and uniform particle size of about 3 microm as determined by the silica support. Nitrogen sorption data confirm that the magnetite-loaded silica particles possess typical mesopore structure with BET surface area of about 183 m(2)/g. VSM data show that the particles display paramagnetic behavior with saturation magnetization of 11.37 emu/g. The magnetic silica microspheres coated with silica shells were tested as adsorbents for rapid extraction of genomic DNA from soybean-derived products. The purified DNA templates were amplified by PCR for screening of genetically modified organisms (GMOs). The preliminary results confirm that the DNA extraction protocols using magnetite-loaded silica microspheres are capable of producing DNA templates which are inhibitor-free and ready for downstream analysis.