Abstract: Objective To investigate the effects of hypoxia / reoxygenation (H/ R) on hippocampal neurons and endoplasmic reticulum (ER) stress in mice. Methods HT22 hippocampal neurons were used for subsequent experiments after H/ R treatment. The effects of H/ R on the proliferation and migration of HT22 hippocampal neurons were detected by Cell Counting Kit-8 and scratch assays and its effects on the morphology of the ER was observed by transmission electron microscopy. The effect of H/ R on the development of HT22 hippocampal neurons was observed by immunofluorescence, and its effects on ER stress in hippocampal neurons were detected by Western blot and quantitative reverse transcription-polymerase chain reaction. Results Compared with the Con group, H/ R inhibited the proliferation and migration of HT22 hippocampal neurons and decreased the expression of the neurodevelopmental protein brain-derived neurotropic factor(P<0. 05). H/ R also reduced the expression of the synaptic plasticity proteins postsynaptic density protein-95 and α-synuclein(P<0. 05). H/ R provoked pronounced ER swelling and up-regulated the key ER-stress markers glucose-regulated protein 94, binding immunoglobulin protein, and activating transcription factor 6, thereby intensifying ER stress(P<0. 05). Conclusions H/ R has a damaging effect on HT22 hippocampal neurons, which may be related to enhanced ER stress.

Abstract: Objective The DNA repair-related protein RAD23B has been implicated in the progression of various malignancies. This study aimed to investigate the role of RAD23B in promoting colorectal cancer ( CRC) metastasis and to elucidate the underlying molecular mechanisms. Methods Cell proliferation, migration, and invasion were assessed using Cell Counting Kit-8 and Transwell assays. A xenograft mouse model was used to evaluate the metastatic potential in vivo. Transcriptomic analysis was carried out by RNA sequencing (RNA-seq) to identify signaling pathways regulated by RAD23B. Expression levels of RAD23B, Talin1, Integrinαv, Integrinβ1, phosphoinositide 3-kinase ( PI3K), phosphorylated PI3K, protein kinase B ( AKT), phosphorylated AKT, and matrix metalloproteinase 9 (MMP9) were measured by Western blot. Results In vitro, overexpression of RAD23B significantly enhanced the proliferation, migration, and invasion abilities of colorectal cancer SW480 and HCT-8 cells (P<0. 05). In the in vivo model, RAD23B overexpression notably increased the number of liver metastatic foci in mice(P<0. 05), indicating that RAD23B promotes the liver metastasis potential of colorectal cancer cells. RNA sequencing revealed that RAD23B overexpression activated cell adhesion, integrin, and PI3K-AKT signaling pathways in SW480 cells ( P<0. 05 ). Western blot analysis demonstrated that RAD23B overexpression upregulated the expression of Talin1, Integrinαv, Integrinβ1, p-PI3K, PI3K, p-AKT, AKT, and MMP9(P<0. 05). Conclusions RAD23B promotes CRC liver metastasis through activation of the Talin1 / Integrin / PI3K/ AKT / MMP9 axis.

Abstract: Objective To evaluate the feasibility of establishing a hypertensive rat model induced by deoxycorticosterone acetate ( DOCA), and to compare the stability of the model between surgery ( unilateral nephrectomy combined with DOCA injection) and non-surgery group (DOCA injection alone). Methods Healthy Wistar rats were divided randomly into surgery group and non-surgery group according to body weight. Rats in surgery group underwent left nephrectomy followed by subcutaneous injection of DOCA (50 mg / kg), five times a week for 5 weeks. Rats in the non-surgery group received DOCA injections following the same protocol for 7 weeks, without nephrectomy. Both groups were given free access to 1% saline solution throughout the experiment. Tail artery blood pressure was monitored weekly until week 9. Blood pressure changes and model stability were compared between the groups. Results Following drug administration, the blood pressure of rats in the surgery group gradually increased. After drug withdrawal, the blood pressure remained persistently elevated with slight fluctuations, resulting in sustained hypertension. During the same period of drug administration and withdrawal, the blood pressure of rats in the nonsurgery group exhibited a consistent change pattern with that in the surgery group, i. e. , a gradual increase after drug administration and sustained maintenance of hypertension after drug withdrawal. Conclusions Subcutaneous DOCA administration can effectively induce hypertension in rats without the need for nephrectomy. Compared with the conventional surgical model, the non-surgical method is simpler, safer, and reproducible, and has broad applicability for hypertension research.

Abstract: Objective To explore the mechanism by which osteoking ( OK) regulates ferroptosis and improves knee osteoarthritis (KOA). Methods We performed Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis by analyzing target genes related to OK, KOA, and ferroptosis through network pharmacology. Human C28 / I2 cells were used to simulate an in vitro arthritis model. Cells were divided into Control, model (interleukin ( IL)-1β 10 mg / L), IL-1β + OK, IL-1β + OK+ advanced glycation end products ( AGEs), IL-1β + Ferrostatin-1( Fer-1) and IL-1β + OK + Fer-1 groups. The inflammatory factor IL-6 and oxidative stress factors malondialdehyde (MDA), superoxide dismutase ( SOD), and glutathione ( GSH) in the cell supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Fe ²⁺levels in each group were detected using cell-specific assay kits. Matrix metalloproteinase ( MMP13 ), cartilage matrix synthesis protein 2 ( COLA2 ), glutathione peroxidase 4 (GPX4), and solute carrier family 7 (SLC) proteins were detected by western blot. A rat model of KOA was created by medial meniscus instability surgery. Rats were divided into sham surgery, model and osteoking groups and treated with the corresponding drugs orally for 8 weeks. Paw-contraction reaction time and weight were measured in each group using a hot plate apparatus and pressure pain gauge, respectively. Serum inflammatory factors tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), IL-6, and the oxidative stress factors MDA,SOD, GSH, and reactive oxygen species (ROS) were detected by ELISA. Knee joints were stained with hematoxylin /eosin, safranin turquoise, and toluidine blue, and the degree of cartilage tissue degeneration was scored according to the OARSI standard. MMP13, COLA2, GPX4, SLC, AGEs, and their receptor (RAGE) proteins were detected in rat cartilage by Western blot. Results Network pharmacology analysis identified the AGEs-RAGE and other signaling pathways as potential key pathways by which OK may act on KOA and ferroptosis. In vitro, MDA, IL-6, and Fe²⁺levels increased in the IL-1 β group ( P<0. 05,P<0. 01), while GSH and SOD levels decreased ( P<0. 05,P<0. 01). MMP13 levels increased ( P<0. 05), while COLA2, GPX4, and SLC levels decreased ( P<0. 01). OK intervention reversed the above result and AGEs weakened the effect of OK, with significant effects on the relevant indicators. OK also enhanced the anti-inflammatory and antioxidant stress effects of the ferroptosis inhibitor Fer-1 on C28 / I2 cells, and improved the metabolic balance. DMM group rats showed significantly increased tenderness,thermal pain, and OARSI score (P<0. 01,P<0. 001), and damage to the cartilage surface and matrix of the knee joint. Compared with the DMM group, OK significantly reduced tenderness, thermal pain, and OARSI scores (P<0. 01), and alleviated damage to the cartilage surface and matrix of the knee joint. COLA2, GPX4, and SLC levels in the articular cartilage were significantly increased in the OK group compared with the DMM group ( P<0. 05), while MMP13, AGEs, and RAGE were decreased ( P<0. 05,P<0. 01). Serum levels of TNF-α, VEGF, IL-6,MDA, and ROS were significantly reduced in the OK group (P<0. 01), while SOD and GSH were increased (P<0. 01). Conclusions OK can inhibit ferroptosis and protect articular cartilage via the AGEs-RAGE axis, thereby exerting anti-arthritis effects.

Abstract: Objective To explore the mechanism by which Sijunzi Tang improves osteosarcopenia (OS) in rats based on the Toll-like receptor 4 ( TLR4) / nuclear factor (NF)-κB pathway. Methods Rats were divided randomly into sham, OS, Sijunzi Tang L, Sijunzi Tang H, estradiol, and Sijunzi Tang H+lipopolysaccharide (LPS) groups. The OS model was constructed by castration combined with intraperitoneal injection of dexamethasone. After treatment with Sijunzi Tang and the TLR4 activator LPS, forelimb muscle strength, whole-body and femoral bone densities, muscle percentage, and distal femoral bone microstructure (percent bone volume(BV/ TV), bone surface density (BS / TV), trabecular number (Tb. N), and trabecular separation degree (Tb. Sp) were measured. The mass fraction of quadriceps muscle was detected, and histopathological examination of femur tissues and quadriceps femoris tissue was carried out by hematoxylin / eosin staining. The cross-sectional area (CSA) of quadriceps muscle fibers was compared. Serum procollagen I N-terminal propeptide ( PINP ), C-telopeptide of type I collagen ( CTX-1 ),osteocalcin (OCN), and inflammatory factors were determined by enzyme-linked immunosorbent assay. TLR4 / NF-κB pathway proteins in the quadriceps femoris and femur tissues were detected by immunohistochemical staining and Western blot. Results In the sham surgery group, OS rats showed decreased forelimb muscle strength, whole-body and femoral bone densities, muscle percentage, BV/ TV, BS / TV, Tb. N, quadriceps mass fraction, muscle fiber CSA, and levels of PINP and OCN (P<0. 05), and increased Tb. SP, levels of CTX-1, interleukin (IL)-6, IL-1β and IL-17, positive expression of TLR4 and phospho (p)-NF-κB p65, TLR4 protein expression, and p-NF-κB p65 / NF-κB p65 (P<0. 05). For the OS group, rats in the Sijunzi Tang L group,Sijunzi Tang H group and estradiol group showed increases in forelimb muscle strength, whole-body and femoral bone densities, muscle percentage, BV/ TV,BS / TV, Tb. N, quadriceps mass fraction and muscle fiber CSA, levels of PINP and OCN (P<0. 05), and decreases in Tb. SP, levels of CTX-1, IL-6, IL-1β and IL-17, positive expression of TLR4 and p-NF-κB p65, TLR4 protein expression and p-NF-κB p65 / NF-κB p65 (P<0. 05). Sijunzi Tang H and estradiol groups, however, had a stronger improving effect on various pathological indicators in OS rats. Rats in the Sijunzi Tang H + LPS group showed decreased forelimb muscle strength, whole-body and femoral bone densities, muscle percentage, BV/ TV, BS / TV,Tb. N, quadriceps mass fraction, and muscle fiber CSA, and decreased levels of PINP and OCN (P<0. 05), and increased Tb. SP, levels of CTX-1, IL-6, IL-1β, and IL-17, positive expression of TLR4 and p-NF-κB p65, TLR4 protein expression and p-NF-κB p65 / NF-κB p65 (P<0. 05). There was no significant difference in various indicators between Sijunzi Tang H group and estradiol group ( P>0. 05). Conclusions Sijunzi Tang can improve muscle atrophy and osteoporosis symptoms in OS rats by suppressing activation of the TLR4 / NF-κB signaling pathway.

Abstract: Objective To preliminarily explore the role of mitochondria-related programmed cell death mechanisms, including apoptosis, autophagy, and ferroptosis, in early-aging cardiac hypertrophy and myocardial fibrosis. Methods Adult male C57BL / 6J mice (5 months old) and early-aging mice (21 months old) ( n= 9 per group) were housed in a specific-pathogen-free environment. Cardiac tissues were collected for micro-computed tomography scanning. Paraffin-embedded sections were stained with hematoxylin / eosin and Van Gieson stain. Cardiac mitochondria were extracted to measure reactive oxygen species (ROS) and expression levels of the dynamics-related proteins dynamin-related protein 1 ( Drp1), mitofusin 2 ( Mfn2), and optic atrophy 1 ( OPA1). Mitochondrial ultrastructure was observed using transmission electron microscopy. Protein expression levels of the inflammatory markers receptor-interacting serine / threonine-protein kinase 3 ( RIP3) and phosphorylated nuclear factor ( p-NF- κB), the fibrosis marker Bcl-2-associated X protein( Bax),collagen type I, the apoptosis markers caspase-3 and cleaved caspase-3, the autophagy markers phosphorylated protein kinase B (p-Akt), protein kinase B (Akt), and microtubule-associated protein 1 light chain 3 I/ II (LC3I/ II), and the ferroptosis markers acyl-CoA synthetase longchain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) were measured by Western blot. Results Compared with adult mice, early-aged mice exhibited myocardial hypertrophy and fibrosis, disordered cardiomyocyte arrangement, and increased mitochondrial ROS levels (P<0. 05). There was no significant change in expression levels of the mitochondrial fission-related protein Drp1, but expression levels of the fusion-related proteins Mfn2 and OPA1 were increased (P<0. 05). Electron microscopy revealed swelling and disordered cristae structures in myocardial mitochondria. Expression levels of the myocardial fibrosis marker Collagen I(P<0. 01), as well as the inflammation-related markers RIP3 and p-NF-κB, were elevated ( P<0. 01). In contrast, expression levels of the autophagy-related protein LC3II were reduced (P<0. 05), while the expression of Bax was increased (P<0. 01),with no significant change in expression of the apoptosis-related protein Cleaved Caspase-3. In addition, expression of the ferroptosis-related protein ACSL4 was increased ( P<0. 001), whereas SLC7A11 expression was decreased ( P<0. 05). Conclusions Early cardiac aging is characterized by hypertrophy and myocardial fibrosis, accompanied by inflammation, mitochondrial dysfunction, and dynamics imbalance. Notably, ferroptosis, rather than apoptosis, emerges as the primary pathway of programmed cell death during the early stages of cardiac aging.

Abstract:The “Healthy China 2030” plan clearly states that “jointly building and sharing, and promoting health for all” is the strategic theme for building a healthy China. It is necessary to promote extensive participation from the whole society, strengthen cross-departmental collaboration, enhance environmental governance, ensure food and drug safety, prevent and reduce harm, and effectively control ecological and social environmental risk factors that affect public health. As a medical institution, it is necessary to focus on deepening comprehensive reforms around the Healthy China strategy, and adopt a broad perspective on education, health, and wellness. In recent years, especially following outbreaks like the severe acute respiratory syndrome outbreak, the “One Health” education concept has emphasized the need for people to coexist harmoniously with other living beings and the environment to achieve true “health”, utilizing new perspectives, heights, and dimensions. As the main channel for disseminating the “One Health” education concept in this new era, colleges and universities need to promote the ideological and political education functions of professional courses, fully implement “One Health” education, and promote the development of values. The basic medical course of “Pathogenic Biology” serves as a bridge connecting core courses of the major degree. This course includes many “ course ideological education ” elements that can be explored. This paper accordingly focuses on how to incorporate the “ One Health” education concept into the ideological and political education system of the “Pathogenic Biology” course.

Abstract:Continuing global research into and the development of hemoglobin-based oxygen carriers (HBOC) has led to increasing attention on ways by which to evaluate their performance in the circulation. This article reviews the preparation method , observation indicators, performance evaluation, and selection suggestions of the main HBOC circulatory performance-evaluation models, including the hemorrhagic shock, isovolumetric blood exchange,circulatory top load, and ischemia-reperfusion models, together with the development of other biomimetic models.

Abstract:Sex hormone levels in women during the menopause are closely related to the progression of Alzheimer’s disease. Elevated levels of follicle-stimulating hormone and decreased levels of estrogen jointly drive the pathological process of Alzheimer’ s disease, including amyloid-β protein deposition, microtubule fibrillary tangles,neuroinflammation, and excessive activation of glial cells. This article reviews the relevant pathways and mechanisms by which the increase in follicle-stimulating hormone levels and deficiency of estrogen during the menopause exacerbate the pathological process of Alzheimer’s disease, providing a reference for subsequent treatment of this disease.

Abstract:Hypoxia is an important part of the pathogenesis of many chronic respiratory and cardiovascular diseases, cerebral ischemia, and solid tumors, and other major diseases. Cell hypoxia models provide an important tool for analyzing the mechanism of hypoxia responses and for screening potential therapeutic targets. The scientific aspects and applicability of model construction, however, can directly affect the reliability of the research conclusion ,and are thus of great significance to the development of related disease prevention and treatment research. Research on the construction and application of cell hypoxia models has currently achieved stage-by-stage result , but there are still some limitations in terms of the model-evaluation system and tissue cell specificity. This paper summarizes the current status of research on the mechanism of hypoxia occurrence and development and the evaluation of indicators. We combine the tissue-specific characteristics, and discuss the construction and optimization of the cellular hypoxia model to provide a reference for future model construction and to support future research, development, and translation of targeted therapies.

Abstract:Parkinson’ s disease ( PD) is a common neurodegenerative disease characterized by progressive degeneration of nigrostriatal dopaminergic neurons and Lewy body morphology. The pathogenic mechanism of PD,however, is not yet fully understood, and the increase in the number of patients places a huge burden on society.Acupuncture, as a distinctive therapy in traditional Chinese medicine, has been widely used to improve motor and non-motor symptoms in patients with PD, possibly acting via a mechanism closely related to the regulation of programmed cell death ( PCD )-related signaling pathways. PCD, including apoptosis, necroptosis, cellular pyroptosis, ferroptosis, and other forms, plays a key role in the process of neuronal damage in PD. This review thus considers the role of PCD in PD. Combined with research progress in acupuncture interventions for PCD, it provides new ideas to clarify the scientific basis of acupuncture treatment for PD, and supports the development of new treatment strategies.

Abstract:Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by alveolar epithelial cell injury, aberrant repair, and pathological fibrosis. Recent studies have identified ferroptosis, as an irondependent, lipid peroxidation-driven form of programmed cell death, as a critical factor in the pathogenesis of IPF.This review elaborates on the fundamental pathological contributions of type Ⅱ alveolar epithelial cells, fibroblasts/ myofibroblasts, and macrophages in IPF, and analyzes the hallmarks of ferroptosis and its association with the characteristic pathological alterations of IPF, namely oxidative stress and dysregulated iron metabolism. Finally, we consider the cellular crosstalk mechanisms from the perspectives of the vicious cycle of paracrine signaling, receptorligand-mediated iron metabolic reprogramming, extracellular matrix-integrin-mediated mechanical stress, and cellular phenotypic reprogramming. A deeper understanding of this intricate cellular interaction network, particularly the pivotal regulatory role of ferroptosis, will provide a crucial theoretical foundation for the development of novel multitarget combination therapeutic strategies.

Abstract:Deep vein thrombosis (DVT) is the third most common cardiovascular disease worldwide but traditional anticoagulation therapy options are limited. The pathophysiology of DVT usually focuses on Virchow’ s triad, but this fails to explain many of the clinical problems. Considering the interactions of metabolic reprogramming with inflammation and coagulation may thus open up new perspectives. This review explores the roles of the metabolic microenvironment in thrombosis and oxidative stress in relation to metabolic reprogramming and thrombosis, analyzes the feedback loop between metabolic reprogramming and inflammatory coagulation, and summarizes the therapeutic strategies of metabolic regulation intervention in DVT, including glucose metabolism, lipid metabolism, and antioxidant therapy. We also highlight the challenges in terms of therapeutic strategies, and indicate potential new approaches to the prevention and treatment of DVT.

Abstract:Macrophages, as intrinsic immune cells, are an important component of the innate immune system and play an important role in immunity against Mycobacterium tuberculosis ( Mtb) infection. After infecting the organism, Mtb mainly parasitizes macrophages, where it can survive and multiply, inducing cell death via apoptosis,autophagy, pyroptosis, ferroptosis, and necroptosis. These different modes of cell death play different roles in the process of Mtb infection. This review considers the different macrophage cell-death pathways and their roles after Mtb infection, with the aim of furthering our understanding of the potential impacts on host immunity during Mtb infection,and providing references for future research into the pathogenic mechanism of Mtb and strategies of anti-tuberculosis treatment.