ObjectiveTo investigate the protective effect of liver-targeted fibroblast growth factor 21 （FGF21） overexpression against CCl₄-induced acute liver injury （ALI） and its association with silent information regulator 1/glutathione peroxidase 4 （SIRT1/GPX4） expression and ferroptosis-related markers. MethodsFemale ICR mice （8 weeks old） were randomly divided into Control， CCl₄， adeno-associated virus （AAV）-FGF21， and AAV-FGF21 + CCl₄ groups. Liver-targeted FGF21 overexpression was achieved via tail vein injection of AAV8-FGF21 virus with green fluorescent protein （GFP） labeling. ALI was induced by intraperitoneal injection of CCl₄ （0.3 mL/kg）， with sample collection at 12 hours. Body weight and liver weight were recorded to calculate the liver weight/body weight. Alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using a biochemical analyzer. FGF21 expression in liver tissues was detected by immunofluorescence. Hepatic histopathology was evaluated by HE staining. RT-qPCR was performed to assess mRNA expression of ferroptosis and iron metabolism-related genes （ACSL4， PTGS2， HJV， FPN， GPX4）， FGF21， and SIRT1. Protein expression of FGF21， SIRT1， and GPX4 were determined by Western blot. Iron content， malondialdehyde （MDA）， glutathione （GSH）， and superoxide dismutase （SOD） activity in liver homogenate were quantified using assay kits. ResultsCompared with the Control group， the FGF21 mRNA and protein expression elevated in CCl₄ group； serum ALT/AST levels and liver weight/body weigh increased. The HE staining results showed that the liver cell membranes were ruptured. The mRNA levels of ACSL4 and PTGS2， the iron content and MDA levels all increased （P<0.01）. The mRNA levels of SIRT1， GPX4， HJV， and FPN， the GSH level and SOD activity all decreased （P<0.05）， and the protein levels of SIRT1 and GPX4 both decreased （P<0.05）. Compared with the CCl₄ group， the serum levels of ALT and AST in the AAV-FGF21 + CCl4 group were decreased （P<0.01）， and the liver/body ratio of mice decreased （P<0.01）. The results of HE staining showed that the degree of liver injury was alleviated. The mRNA levels of ACSL4 and PTGS2， iron content and MDA level all decreased （P<0.01）， while the mRNA levels of SIRT1， GPX4， HJV， FPN， GSH level and SOD activity all increased （P<0.05）. The protein levels of SIRT1 and GPX4 both increased （P<0.05）. ConclusionFGF21 overexpression is associated with enhanced SIRT1/GPX4 expression and suppression of ferroptosis features， thereby contributing to the alleviation of CCl₄-induced ALI.

Objective To investigate the effect of berberine and its mechanism in delaying silica-induced lung injury via network pharmacology and molecular docking, followed by verification with animal experiments. Methods i) Targets of berberine and silicosis were obtained from multiple databases, and the common targets were identified. A protein-protein interaction network was constructed using the STRING database, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the common targets via the DAVID database. Molecular docking was conducted using AutoDock software for validation. ii) Specific pathogen-free Wistar rats were randomly divided into five groups, with six rats in each group. Rats in the model group, pirfenidone group, and low- and high-dose berberine groups received a single non-exposure intratracheal instillation of 1.00 mL silica suspension at a mass concentration of 100 g/L, whereas rats in the control group received an equal volume of 0.9% sodium chloride solution. Rats in the pirfenidone group were administered with pirfenidone solution at a dose of 100 mg/kg body weight, and rats in the low- and high-dose berberine groups were administered with berberine solutions at doses of 100 and 200 mg/kg body weight, respectively. The remaining two groups received equal volumes of 0.9% sodium chloride solution. All treatments were administered by gavage once daily for 28 consecutive days from 24 hours after silica exposure. After intervention, rats in each group were sacrificed. Histopathological changes in lung tissues were observed, and the lung organ coefficient was detected. Serum malondialdehyde (MDA) was detected by colorimetry. Serum inflammatory cytokine were detected by enzyme-linked immunosorbent assay. The expression of fibrosis markers in lung tissues was detected by immunohistochemistry. The protein expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and phosphorylated AKT (p-AKT) in lung tissues was determined by Western blot. Results i) Network pharmacology analysis identified 46 potential targets. The core targets included AKT1, tumor protein p53, caspase-3, matrix metalloproteinase-9, and albumin. GO enrichment analysis revealed that the targets related to the anti-fibrotic effect of berberine against silicosis were mainly enriched in biological processes including signal transduction, negative regulation of apoptosis, extracellular matrix degradation, and response to reactive oxygen species. KEGG enrichment analysis showed that berberine exerted anti-silicosis effects through multiple signaling pathways, such as the PI3K/AKT pathway, hypoxia-inducible factor-1 pathway, and advanced glycation end product (AGE)-receptor for AGE pathway. ii) Histopathological examination revealed that rats in the model group presented typical pulmonary fibrotic lesions. Pathological lung injury of rats was alleviated in all three intervention groups (pirfenidone group, low- and high-dose berberine group) compared with the model group. The improvement in the high-dose berberine group was superior to that in the low-dose berberine group and was comparable with that in the pirfenidone group. rats in the model group showed increased lung organ coefficient, serum MDA, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) (all P<0.05), as well as increased expression of typeⅠcollagen (COLⅠ), α-smooth muscle actin (α-SMA) and PI3K, and p-AKT/AKT ratio in lung tissues (all P<0.05) when compared with the control group. The above indicators were lower in rats in both the low- and high-dose berberine groups than those in the model group (all P<0.05). Serum MDA, IL-1β, and TNF-α, as well as relative expression of COLⅠ and α-SMA in lung tissues, were lower in the high-dose berberine group than in the low-dose berberine group (all P<0.05). Conclusion Berberine may alleviate pulmonary inflammation and fibrosis in rats through regulation of the PI3K/AKT signaling pathway, thereby delaying the progression of silicosis.

ObjectiveTo investigate the effects of modified Ditan tang on genes related to the transcription-translation feedback loop （TTFL） of biorhythm in the rat model of non-alcoholic fatty liver disease （NAFLD） and its mechanism for prevention and treatment of NAFLD. MethodsSixty-five healthy SPF male SD rats were randomly assigned into blank （n=20）， model （n=15）， and low-， medium-， and high-dose （2.68， 5.36， and 10.72 g·kg^-1·d^-1， respectively） modified Ditan tang （n=10） groups. Other groups except the blank group were fed a high-fat diet for 12 weeks. The modified Ditan tang groups were treated with the decoction at corresponding doses by gavage， and the blank and model groups were treated with an equal volume of normal saline from the 9th week for 4 weeks. The levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） in the serum were measured by an automatic biochemical analyzer. TG and non-esterified fatty acid （NEFA） assay kits were used to measure the levels of TG and NEFA in the liver. The pathological changes in the hypothalamus and liver were observed by hematoxylin-eosin staining， and the lipid deposition in the liver was observed by oil red O staining. The levels of brain-muscle ARNT-like protein 1 （BMAL1/ARNTL） in the hypothalamus and liver were determined by immunohistochemical staining. The mRNA and protein levels of BMAL1， circadian locomotor output cycles kaput （CLOCK）， period circadian clock 2 （PER2）， and cryptochrome1 （Cry1） in the hypothalamus and liver were determined by Real-time PCR and Western blot， respectively. ResultsCompared with the blank group， the model group showed elevated levels of TG， TC， LDL-C， AST， and ALT （P<0.01） and a lowered level of HDL-C （P<0.05） in the serum， elevated levels of TG and NEFA in the liver （P<0.01）， pyknosis and deep staining of hypothalamic neuron cells， and a large number of vacuoles in the brain area. In addition， the model group showed lipid deposition in the liver， up-regulated mRNA and protein levels of CLOCK and BMAL1 （P<0.01）， and down-regulated mRNA and protein levels of Cry1 and PER2 （P<0.01） in the hypothalamus and liver. Compared with the model group， all the three modified Ditan tang groups showed lowered levels of TG， TC， LDL-C， ALT， and AST （P<0.05， P<0.01） and an elevated level of HDL-C （P<0.05） in the serum， and lowered levels of TG and NEFA （P<0.05， P<0.01） in the liver. Furthermore， the three groups showed alleviated pyknosis and deep staining of hypothalamic neuron cells， reduced lipid deposition in the liver， down-regulated mRNA and protein levels of CLOCK and BMAL1 （P<0.05， P<0.01）， and up-regulated mRNA and protein levels of Cry1 and PER2 （P<0.05， P<0.01） in the hypothalamus and liver. ConclusionModified Ditan tang can reduce lipid deposition in the liver and regulate the expression of CLOCK， BMAL1， Cry1， and PER2 in the TTFL of NAFLD rats.

Plant exosome-like nanovesicles(PELNs)are extracellular nanoparticles(30 to 300 nm in diameter)secreted by plant cells,characterized by a unique lipid-protein-nucleic acid compo-sition and functional attributes.These vesicles play pivotal roles in plant metabolism,immune de-fense,and the regulation of cellular physiological activities across species.This review systematically summarized recent research advances in PELNs,encompassing their biogenesis pathways,composi-tional analysis,extraction/isolation protocols,storage stability,identification methodologies,and ap-plications in skeletal system disorders.Critical challenges and future research directions in this field also discussed,providing valuable references for subsequent investigations.

AIM To investigate the protective effects and mechanisms of aqueous extract of Cimicifugae Rhizoma on intestinal mucosa in a mouse model of ulcerative colitis(UC).METHODS The UC mouse models established by sodium dextran sulfate were allocated into different groups and administered with sulfasalazine(200 mg/kg)or aqueous extract of Cimicifugae Rhizoma(3.9,7.8 g/kg)by gavage,respectively.The mice had their changes of body weight,defecation patterns,disease activity index(DAI)and colon length recorded;their colon tissue pathological alterations and goblet cell quantification analyzed through HE and AB-PAS staining;their ROS levels in colon tissue measured via ELISA;their mRNA expressions of inflammatory cytokines,Nrf2/HO-1 signaling pathway components and NLRP3/Caspase-1/GSDMD pathway regulators in colon tissue assessed by RT-qPCR;their protein expressions of Nrf2/HO-1 and NLRP3/Caspase-1/GSDMD pathway verified by immunohistochemistry;and their ZO1 and Occludin tight junction proteins in colon tissues quantified by Western blot analysis.RESULTS Compared to the model group,the high-dose Cimicifugae Rhizoma aqueous extract group demonstrated significantly increased body weight,colon length and DAI scores(P＜0.01);mitigated intestinal mucosal barrier damage;reduced ROS levels in colon tissue(P＜0.01);suppressed mRNA levels of pro-inflammatory factors IL-1β,IL-6 and TNF-α in colon(P＜0.01);elevated expressions of tight junction protein ZO1 and Occludin in colon tissue(P＜0.05);upregulated mRNA and protein expressions of Nrf2,NQO1 and HO-1 in colon tissue(P＜0.05,P＜0.01);downregulated mRNA and protein expressions of Keap1(P＜0.05);and reduced expressions of NLRP3 inflammasome components(ASC,Caspase-1,GSDMD)in mRNA and protein(P＜0.05,P＜0.01).CONCLUSION The aqueous extract of Cimicifugae Rhizoma exerts protective effects against UC through dual mechanisms involving redox regulation and pyroptosis inhibition by reducing ROS level via Nrf2/HO-1 pathway activation and attenuating NLRP3-mediated pyroptosis via Caspase-1/GSDMD pathway inhibition,and thereby synergistically preserves the structural and functional integrity of intestinal mucosal barrier and mitigates UC progression.

AIM To analyze the dynamic change patterns of aflatoxin B1,aflatoxin B2,aflatoxin G1,aflatoxin G2,T-2 toxin and deoxynivalenol contents during the fermentation of Massa Medicata Fermentata.METHODS The analysis was performed on a 40 ℃ thermostatic Waters ACQUITY UPLC HSS T3 column(100 mm×2.1 mm,1.8 μm),with the mobile phase comprising of 0.01％formic acid-[acetonitrile-methanol(1∶1)]flowing at 0.3 mL/min,and electron spray ionization source was adopted in positive ion scanning with multiple reaction monitoring mode.RESULTS Six mycotoxins showed good linear relationships within their own ranges(R2＞0.998 0),whose average recoveries were 76.1％-119.3％with the RSDs of 0.49％-9.27％,and except for deoxynivalenol,their contents demonstrated the trends of growing out of nothing and gradually increasing.CONCLUSION The risk of mycotoxin infection exists in the fermentation of Massa Medicata Fermentata.This simple,efficient,rapid and sensitive method can provide a reference for whole-process monitoring the fermentation process for Massa Medicata Fermentata.

The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective:To assess the physical activity levels(PA)and patterns among older adults in rural areas, as well as to evaluate the association between PA levels and the conditions of sarcopenia and frailty.Methods:A cross-sectional survey was conducted involving 690 rural individuals aged 60 and above.Data on socio-demographic characteristics were collected, while Appendicular Skeletal Muscle Mass Index(ASMI)and grip strength were measured.The Physical Activity Scale for the Elderly(PASE)was employed to evaluate PA levels.Based on their PASE scores, participants were categorized into three groups: low PA level, medium PA level, and high PA level.Sarcopenia was defined according to the 2019 criteria established by the Asian Working Group for Sarcopenia(AWGS), and frailty status was assessed using the Frail Scale.Results:Among the study sample, 38.0% exhibited a low level of physical activity, 46.2% had a medium level, and 15.8% engaged in high levels of activity.As age increased, the level of physical activity among older adults significantly declined.Most of the physical activity reported by participants was attributed to household chores and farming-related activities.Participation in structured exercise among older adults was notably low, with only 1.3% engaging in muscle-strengthening exercises on a weekly basis.The majority(53.9%)reported walking as their preferred form of weekly exercise.After adjusting for confounding factors, the prevalence of sarcopenia was found to be 0.40(95% CI: 0.26-0.62)times lower in the medium physical activity group, and 0.56(95% CI: 0.31-1.01)times lower in the high physical activity group, compared to those with a low physical activity level.Similarly, regarding frailty as a negative outcome, the prevalence was 0.66(95% CI: 0.51-0.84)times lower in the medium physical activity group and 0.46(95% CI: 0.30-0.73)times lower in the high physical activity group, relative to the low physical activity group.When using PASE scores as a continuous variable, the results remained consistent. Conclusions:The physical activity levels of rural older adults are inadequate, and participation in multicomponent exercise programs is notably low.A lower level of physical activity is significantly associated with a higher prevalence of sarcopenia and frailty.Our findings indicate that it is essential to implement physical activity education and interventions to enhance exercise health literacy and to prevent sarcopenia and frailty among rural older adults.