Abstract: Objective　N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B) and its phosphorylation are involved in cerebral ischemia/hypoxic neural injury. Hypoxic preconditioning (HPC) can serve as an endogenous protective intervention to protect the brain from ischemic/hypoxic injury. This study intended to explore the effect of HPC on NR2B and the phosphorylation of its two tyrosine sites (1252 and 1336) in hippocampal cells through in vivo and in vitro experiments and thus determine the role of NR2B in HPC neuroprotection. Methods　6~8 weeks-old male SPF-grade ICR mice and the mouse hippocampal neuron cell line HT22 were repeatedly exposed to hypoxia to replicate HPC animal and cell models. Western blot and immunofluorescence were applied to detect the levels of NR2B and the phosphorylation levels of its tyrosine 1336 (pY1336NR2B) and 1252 (pY1252NR2B) residues in the hippocampus of mice and HT22 cells. The distributions of NR2B, pY1336NR2B, and pY1252NR2B in the synaptic site (TxP) and extrasynaptic site (TxS) were analyzed by Western blot. The levels of cleaved caspase-3 and α-spectrin, which indicate cell apoptosis, were also detected. Results　HPC downregulated the levels of NR2B and pY1336NR2B in the mouse hippocampus and HT22 cells. Changes in NR2B and pY1336NR2B levels in the TxS of the mouse hippocampus were similar to those in hippocampus and HT22 cells, whereas changes in the TxP showed the opposite trend. Conclusions　Downregulation of NR2B and pY1336NR2B may be involved in HPC-induced neuroprotection, and their localization at synapses and extrasynapses may play different roles in neuroprotection.

Abstract: Objective　To investigate the inhibitory effect of Boschniakia rossica boschnaloside (BRBN) on the epithelial-mesenchymal transition (EMT), invasion, and migration of human hepatoma SMMC-7721 cells. Methods Transforming growth factor β₁ (TGF-β₁) was used to induce an EMT model of SMMC-7721 cells. The cells were divided into a control group, model group, and BRBN group. Cell migration was detected with a wound healing test, and cell invasion was observed by Transwell chamber assay. The expression of E-cadherin, N-cadherin, Vimentin, Slug, Twist1, ZEB1, and signal transducers and activators of transcription 3 (STAT3) were determined with the Western blot method. Results TGF-β₁ resulted in spindle-shaped SMMC-7721 cells, down-regulated the expression of E-cadherin, and upregulated the expression of N-cadherin and vimentin, suggesting that SMMC-7721 cells obtained a mesenchymal phenotype. Compared with the model group, the BRBN group SMMC-7721 cells’ expression of E-cadherin was significantly increased, their expression of N-cadherin and vimentin were reduced, and their expression of Slug, Twist1, and ZEB1, as well as STAT3 phosphorylation, were down-regulated. Conclusions　BRBN inhibits the invasion and migration of human hepatoma SMMC-7721 cells, possibly by reversing EMT through the STAT3 pathway.

Abstract: Objective　To investigate the effects of combined exposure to PM2.5 and a high-salt diet on hepatic inflammation and lymphangiogenesis in mice. Methods　Thirty-two C57BL/6J mice were assigned randomly to control, PM2.5, high-salt(HSD), and PM2.5+HSD groups. Mice in the HSD and PM2.5+HSD groups were fed an 8% high-salt diet for 8 weeks, while mice in the other groups were fed a control diet containing 0.4% salt. Mice in the PM2.5 and PM2.5+HSD groups were treated with PM2.5 by tracheal instillation (twice per week), and mice in the other groups were instilled with equal volumes of saline (twice per week). All mice were sacrificed after the last PM2.5 exposure. Tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 levels in the liver tissues of the mice were determined and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE1) expression in the liver tissues was visualized by immunofluorescence staining. Protein expression levels of the lymphangiogenesis markers PROX1 and LYVE1, and the lymphangiogenesis regulatory proteins vascular endothelial growth factor (VEGF) receptor (VEGFR) 3 and VEGF-C in liver tissue were measured by Western blot. Results　TNF-α and IL-6 levels and protein expression levels of PROX1, LYVE1, VEGFR-3, and VEGF-C in liver tissues were increased in the HSD group compared with the control group (P<0.05), and in the PM2.5+HSD group compared with the HSD group (P<0.05). Moreover, there were significant interaction effects between PM2.5 and a high-salt diet on these above indicators. Conclusions　Combined exposure to PM2.5 and a high-salt diet aggravated hepatic inflammation and may increase hepatic lymphangiogenesis by upregulating VEGFR-3 and VEGF-C in liver of mice.

Abstract: Objective　This study aims to evaluate the therapeutic effect of Gualou Xiebai Banxia Decoction (GXBD) on an atherosclerosis (AS) mouse model and explore its underlying mechanism. Methods　Male ApoE^-/- mice were fed a high-fat diet for 16 weeks to induce the AS model. Mice were divided into a model group, control group, GXBD treatment groups (at different doses), and a lipid-lowering drug group. Histological assessments (HE and Oil Red staining) were performed to observe arterial changes. MicroRNA (miRNA) sequencing was used to analyze the regulatory effect of GXBD. Western blot and immunofluorescence were applied to confirm key protein expression. Results　GXBD significantly reduced plaque formation and regulated several AS-related miRNAs, notably impacting Epsin1 and FGFR1 protein expression. Western blot and immunofluorescence further validated these effects. Conclusions　GXBD demonstrated a therapeutic effect on AS by regulating miR-3102-5p, miR-3547-5p, and miR-7080-5p, downregulating Epsin1, and upregulating FGFR1, thereby inhibiting endothelial-to-mesenchymal transition.

Abstract: Objective　To study the effect of DNA methyltransferase 1 (DNMT1) on sm-like protein-4 (LSM4) in hepatocyte apoptosis in mice induced with Hcy. Methods　12 ApoE^-/- mice were divided into two groups: normal diet (ND, n=6) and high methionine diet (HMD, n=6) groups. Normal hepatocytes of NCTC1469 were divided into a normal group (control, 0 μL/L Hcy), Hcy intervention group (Hcy, 100 μL/L Hcy), NC siRNA-transfected control group (si NC group, 0 μmol/L Hcy), LSM4 siRNA-transfected group (si-LSM4 group, 0 μmol/L Hcy), DNMT1 siRNA transfected group (si-DNMT1 group, 0 μmol/L Hcy), NC siRNA-transfected Hcy intervention group (Hcy+si-NC group, 100 μmol/L Hcy), LSM4 siRNA-transfected Hcy intervention group (Hcy+si-LSM4 group, 100 μmol/L Hcy), and DNMT1 siRNA-transfected Hcy intervention group (Hcy+si-DNMT1 group, 100 μmol/L Hcy). Analysis of the expression of LSM4 in various tissues was conducted using the NCBI database. Quantitative real-time PCR (qRT-PCR) and Western blot were used to detect differences in LSM4 protein expression in mouse tissues (HMD and ND) and hepatocytes (control and Hcy). Western blot was used to detect the expression of Bcl2-associated X (Bax) and B-cell lymphoma-2 (Bcl-2). The cell apoptosis rate in the Control, Hcy, Hcy+si-NC, and Hcy+si-LSM4 groups were detected by flow cytometry. MethPrimer online software was used to analyze the CpG islands of LSM4 promoter region. The expression of LSM4 in the Hcy+si-DNMT1 group was detected by qRT-PCR and Western blot. Results　The expression of LSM4 in HMD,Hcy group was higher than that in the ND and Control group (P<0.05). Bax protein expression was significantly higher, but Bcl-2 was significantly lower in Hcy group compared with those of the Control group (P<0.05). The expression of Bax protein was significantly lower, but the level of Bcl-2 was significantly higher in the Hcy+si-LSM4 group compared with those in the Hcy+si-NC group (P<0.05). The cell apoptosis rate in the Hcy group was higher than that in the Control group (P<0.05), while the apoptosis rate in the Hcy+si-LSM4 group was lower than that in the Hcy+si-NC group (P<0.05). MethPrimer database analysis showed that the promoter region of LSM4 was GC-rich, and there was one CpG island. Compared with the Hcy + si-NC group, the Hcy+si-DNMT1 group’s expression of LSM4 protein was increased (P<0.05). Conclusions　DNMT1 regulates LSM4 hypomethylation to increase its expression, thereby promoting Hcy-induced apoptosis of mouse hepatocytes.

Abstract: Objective　We generated ob mice (C57BL/6N-Lep^em1 /Nifdc) with Lep gene knockout (ob/ob) using the CRISPR/Cas9 system, to establish a suitable animal model for preclinical drug evaluation for clinical diseases such as diabetes. Methods　According to the principle of CRISPR/Cas9 target design, single guide RNA targeting the mouse Lep gene was designed for transcription in vitro, and microinjected with Cas9 mRNA into mouse zygotes. Mouse tail DNA was extracted and detected by polymerase chain reaction and sequencing, followed by mating of positive and wild-type mice. Blood biochemistry and liver pathology were assessed in homozygous ob mice. Results　Eight positive mice were identified and a stable mouse strain was selected for further breeding. Serum triglycerides, total cholesterol, and alanine aminotransferase levels were significantly higher in homozygous ob mice than in wild-type mice, and liver pathology showed inflammatory infiltration and lipid vacuolar transformations. Conclusions　We successfully established a Lep gene knockout mouse model, which will provide an important addition to the national rodent experimental animal database and an animal model for preclinical drug evaluation.

Abstract: Objective　To investigate the combined effect of CdCl₂ and nicotine on the cell cycle and apoptosis of mouse spermatogonium (GC-1). Methods　A CCK-8 assay was used to detect the inhibition of GC-1 cell proliferation in the CdCl₂ groups, nicotine groups, and combined groups at 24 hours. Flow cytometry was used to detect the effects of CdCl₂, nicotine, and the combined treatment on the cell cycle and apoptosis of GC-1 cells at 24 hours. Results　The inhibitory effect on the proliferation of GC-1 cells increased with the rise in CdCl₂ and nicotine concentrations. The IC₅₀ values of CdCl₂ and nicotine alone were 5.409 μmol/L and 2814 μmol/L, respectively. The IC₅₀ values of CdCl₂ and nicotine combined for GC-1 cells were 4.422, 4.532, 3.309, and 2.532 μmol/L at nicotine concentrations of 0.175, 0.350, 0.700, and 1.400 mmol/L, respectively, thus there was an obvious decrease with nicotine concentration. The IC₅₀ values were lower than those of the group administered CdCl₂ alone, and the combined action of CdCl₂ and nicotine had synergistic effects. Cell cycle result showed that CdCl₂ concentration of 2.5 μmol/L increased the percentage of G2 /M phase cells in the combination groups. Cells were arrested in the G2 /M phase in the combined groups (P<0.01). When combined, CdCl₂ and nicotine had a synergistic effect on the cell cycle, and the synergistic effect of CdCl₂ was stronger. The result of apoptosis showed the proportion of cells in apoptosis increased in the 2.5 μmol/L CdCl₂ group. Compared with the CdCl₂ and nicotine single-treatment groups, the CdCl₂ and nicotine combined group’s percentage of cells in apoptosis increased significantly(P<0.01), and CdCl₂ had a stronger synergistic effect. Conclusions　Combinations of CdCl₂ and nicotine had a synergistic effect on mouse spermatogonium(GC-1)and led to enhanced cytotoxicity, G2 /M cell cycle arrest, and cell apoptosis. CdCl₂ was the main effect factor in the synergistic effect.

Abstract: Objective　To establish different sleep deprivation models in zebrafish to provide reproducible and practical modeling reference solutions for basic research on insomnia. Methods　Zebrafish insomnia models were induced by two interventions: continuous light (150 Lux) and light plus caffeine. The zebrafish were divided randomly into control, light, caffeine(100 μmol/L), and combined light and caffeine groups. The locomotor ability of zebrafish in each group was observed using open field and circadian rhythm behavioral experiments. The expression and secretion of related sleep genes and the neurotransmitter 5-hydroxytryptamine (serotonin; 5-HT) were detected using quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results　Sleep time, resting time (during the day), and sleep rounds were significantly reduced (P<0.01 or p=0.01) and the distance traveled was significantly increased in the light group compared with the control group (P<0.01). The resting time (daytime) and sleep rounds were increased in the combined and caffeine groups (P<0.01) compared with the control group. There was no significant difference in the activity distance between the combined and caffeine groups (P>0.05). The percentages of swimming distance and swimming time in the central area were decreased in the light group compared with the control group (P<0.05), and were both decreased in the caffeine group compared with the light group (P<0.01). HT receptor 1Aa (HTR1aa) mRNA expression at 6:00. and 12:00 was up-regulated in the light group compared with the control group (P<0.05), but there was no significant difference in HTR1ab mRNA levels between the light group and the combined group (P>0.05). 5-HT secretion was decreased in the light group at 6:00 (P<0.01) and at 12:00 compared with the control group. 5-HT levels were reduced in both the light and combined groups (P<0.01), and secretion levels in the light and combined groups were still lower than in the control group at 18:00. (P<0.01). Conclusions　Light alone is the best intervention for modeling long-lasting insomnia in zebrafish larvae. The responsible mechanisms may be related to the HTR1aa gene as well as biological factors such as 5-HT.

Abstract: Objective　To explore the feasibility of confirming pre experimental conditions in Mongolian gerbils using three-dimensional time-of-flight magnetic resonance angiography (3D-TOF-MRA). Methods　The cerebral arteries in Monglian gerbils were imaged using a 7.0T magnetic resonance imaging scanner and the data were processed using RadiAnt DICOM Viewer software. The imaging result were then validated and compared using the latex perfusion method, and 3D reconstruction of the posterior communicating branch and surrounding cerebral blood vessels were performed using analytical software. Finally, we attempted to use this method to screen for abnormal vascular development in Mongolian gerbils, to verify the effectiveness of this method. Results　3D-TOF-MRA effectively demonstrated the cerebral arteries in live Mongolian gerbils. The technique showed high accuracy for observing the main cerebral arteries in Mongolian gerbils, but its ability to show finer vascular branches was not as good as the latex perfusion method. The data obtained using 3DTOF-MRA could be used for 3D reconstruction of blood vessels, and the technology could be applied to screen for abnormal arterial structures in Mongolian gerbils. Conclusions　3D-TOF-MRA technology can be applied for the structural observation and related research of the cerebral arteries in live Mongolian gerbils.

Abstract:Alzheimer’s disease (AD) is a degenerative neurological disorder that can lead to cognitive decline, mental behavior abnormalities, and a reduced ability to undertake daily life activities. Tryptophan is an essential amino acid for the human body and is produced by three main metabolic pathways, namely kynurenine, 5-hydroxytryptamine, and indole derivatives. Influencing the metabolites of tryptophan can ameliorate neuroinflammation in the brain and significantly improve cognitive ability, while the occurrence and development of AD are reduced. In this paper, we review the research literature on the use of tryptophan metabolism intervention in AD in the last 3 years from CNKI, PubMed, and other databases, and summarize its mechanism of action, with a view to providing a reference for further research on anti-AD drugs.

Abstract:Traumatic brain injury (TBI) is caused by the direct or indirect effects of external factors that result in structural or functional loss of brain tissue. Astrocytes are homeostatic cells in the central nervous system that proliferate and activate rapidly in the early stages of TBI. They then participate in a series of pathological processes, such as neuroinflammation, blood-brain barrier disruption, glial scarring, and excitotoxicity after injury, and thus play a crucial role in secondary neurological injury following TBI. This paper reviews the role of astrocytes in the repair of TBI damage, with the aim of providing new strategies for the prevention and treatment of TBI.

Abstract:Volume-regulated anion channels (VRACs) are present in vertebrate cells and a variety of tumor cells. VRACs include leucine-rich-repeat-containing 8A (LRRC8A) and its four homologous family members (LRRC8B-E), of which LRRC8A is an essential subunit. It has been confirmed that the VRAC LRRC8A is involved in the proliferation, migration, invasion, and multi-drug resistance of tumor cells through various signaling pathways. This ion transporter has shown good potential for use in strategies to kill tumor cells and prevent the development of tumors and can be used as a new target for tumor therapy. Therefore, this paper reviews the latest research on the involvement of LRRC8A in tumorigenesis and development. The molecular structure, function, and regulation of LRRC8A in tumor and immune cells, with emphasis on targeting LRRC8A in tumor diagnosis and immunotherapy, are discussed, providing a reference for studies exploring LRRC8A as a new tumor therapy target.

Abstract:Abdominal aortic aneurysm (AAA) is a hidden and fatal disease, but its underlying developmental mechanism remains unclear. Phenotypic switching of vascular smooth muscle cells and pyroptosis have been identified as biological processes closely related to the appearance and progression of AAA, with potentially important roles in the mechanism of AAA and in providing new directions for its diagnosis and treatment. In this review, we discuss phenotypic switching of vascular smooth muscle cells and the regulatory relationship between cell pyroptosis and AAA.

Abstract:The incidence of neurodegenerative diseases (NDs) has been increasing. Current drugs targeting NDs are mainly based on relieving symptoms rather than effecting a cure, and fail to prevent disease progression. Exosomes are a subset of extracellular vesicles that can be produced by almost all cells in the human body and exert biological effects in a variety of ways after their release into the extracellular environment. Many studies have shown that the unique biological characteristics of exosomes mean that they can be used as a tool for the prevention, remission, treatment and diagnosis of NDs. This article reviews the biogenesis of exosomes and their roles in ND, to provide new ideas for the clinical treatment of these conditions.

Abstract:Peripheral nerve injury (PNI), caused by contusions, fractures, and other traumas, may lead to abnormal sensory function, limited motor capabilities, neuropathic pain, and muscle atrophy, which can severely impact the patient’s quality of life. Post-PNI, Wallerian degeneration occurs, involving axonal degeneration and myelin sheath collapse. Notably, the dorsal root ganglia (DRG), as the primary sensory neurons in pain signal transmission, are crucial targets in neuropathic pain (NP) induced by PNI, and changes in these targets trigger a series of complex signaling pathway alterations. Among these, the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, as a vital regulator of cell survival and death, promotes Schwann cell proliferation and migration, thereby enhancing axonal growth and myelination to facilitate nerve regeneration, and supports the survival of DRG neurons to alleviate NP. Current treatment method, including stem cell transplantation and neurotrophic medications, all have certain limitations. Traditional Chinese medicine (TCM) has the advantages of low cost and few side effects, and is widely used for the treatment of PNI. This article reviews the relationship between oxidative stress, apoptosis, autophagy, inflammation, angiogenesis, the cell cycle, and other pathophysiological mechanisms in PNI and the PI3K/Akt signaling pathway, as well as its associated molecular targets. We also discuss the potential mechanisms of action of TCM monomers, compound formulas, and acupuncture based on the PI3K/Akt signaling pathway in the treatment of PNI, aiming to provide systematic and standardized theoretical guidance for the healing of PNI with TCM and a useful reference for the development of related medications.

Abstract:Polycystic ovarian syndrome (PCOS) is a prevalent reproductive endocrine metabolic disorder within obstetrics and gynecology. Two pivotal features of PCOS, insulin resistance(IR) and hyperandrogenemia(HA), are both instrumental in its pathogenesis. There exists a discernible correlation between PCOS and mitochondrial oxidative stress, a relationship heavily influenced by various factors, most notably IR and HA. This article endeavors to elucidate the interplay between IR, HA, and mitochondrial oxidative stress while also exploring the impacts of insulin resistance and hyperandrogenemia on mitochondrial oxidative stress among PCOS patients.

Abstract:Hypoxia is associated with the occurrence and development of many diseases in clinical settings. Cell hypoxia not only serves as a vital marker for disease advancement, but also plays a pivotal role in exacerbating the disease process, and improving tissue hypoxia may thus provide new strategies for the treatment of related diseases. Further investigation of these diseases at the cellular and molecular levels requires the establishment of a cellular hypoxia model. Current extensively employed hypoxic cell models can be categorized primarily into three types: chemical hypoxia, physical hypoxia, and glucose deprivation hypoxia models. This article reviews the various types of hypoxic cell models and scrutinizes their applications and limitations in disease research.

Abstract:Osteoarthritis (OA) is a chronic degenerative disease with a high incidence rate among middle-aged and elderly people. It can cause joint pain, deformity, and functional impairment, leading to a heavy burden on patients and their families. Peroxisome proliferator-activated receptor γ (PPARγ) is a recently discovered ligand-dependent transcriptional regulatory factor that can regulate fat and carbohydrate metabolism, inflammation, and immune processes. Research has shown that the PPARγ gene plays an important role in OA cartilage degeneration, synovitis inflammation, and adipose lesions, and can affect OA progression through signaling pathways such as the AMP-activated protein kinase, Wnt, and sirtuin 1 pathways. This review focuses on the role and mechanism of the PPARγ gene in OA, in relation to jointrelated tissues and key signaling pathways.

Abstract:Parkinson’s disease (PD) is a highly complex neurodegenerative disease, and its pathogenesis is influenced by DNA methylation, histone modification, and non-coding RNA. Research on Parkinson’s brain tissue and blood has shown that changes in DNA methylation/histone modification levels and variations in ncRNA and its target genes may trigger neurodegeneration changes in the brain and serve as potential non-invasive biomarkers for PD. Exercise can ameliorate the neurodegenerative diseases caused by aging and PD by reversing epigenetic changes. This article reviews the role of epigenetics in the pathogenesis of PD, explores the benefits of exercise, and provides a theoretical reference for research into improving PD by regulating epigenetics through exercise.

Abstract:Microglia efferocytosis, the process by which microglia phagocytose damaged and dead cells, has anti inflammatory and pro-damage repair effects. Recent studies have shown that microglia efferocytosis plays a crucial role in the pathogenesis of Alzheimer’s disease (AD) and may be a novel therapeutic target for AD. This paper reviews the relationship between microglia efferocytosis and AD pathogenesis and the potential of using efferocytosis-related molecules as therapeutic targets for AD. The aim of this review is to provide new ideas and approaches for the treatment of AD.

Abstract:MicroRNAs (miRNAs) comprise a class of endogenous RNA molecules with a typical length of 19~25 nucleotides. They regulate gene expression levels by identifying homologous sequences and intervening in transcription, translation, or epigenetic processes. miRNAs have potential applications in relation to the pathogenesis, progression, and treatment of deep vein thrombosis (DVT). DVT refers to the abnormal coagulation of blood within the lumen of the deep veins, blocking the venous lumen and obstructing the venous return, especially in the lower limbs. Furthermore, detachment of the thrombus and entry into the lungs can lead to death. This article comprehensively reviews recent research findings regarding the diverse mechanisms of action of miRNAs in relation to DVT. Given that the regulation of miRNA expression using targeted therapeutic approaches may promote the recovery of DVT, this article also discusses the potential applications of miRNAs for the clinical diagnosis and treatment of DVT, and aims to provide valuable references and insights for future clinical and basic research in the field of DVT.