Abstract: Objective To investigate the regulatory mechanisms of a disintegrin and metalloproteinase 17 (ADAM17) on endoplasmic reticulum stress (ERS) in cyclosporine A (CsA)-induced renal fibrosis (RF), and to identify potential novel therapeutic targets in the progression of RF. Methods A CsA-induced RF model was established in SD rats through gavage. Plasmid transfection and siRNA were employed to overexpress or knock down ADAM17 in vivo. Indices including body weight, urine protein / creatinine ratio (UPCR), serum blood urea nitrogen (BUN), serum creatinine ( SCR), cystatin C (Cys-C), transforming growth factor-β1 ( TGF-β1), and ADAM17 were measured. Pathological changes in kidney tissues were observed with hematoxylin-eosin staining and Masson trichrome staining. Ultrastructural alterations were examined using transmission electron microscopy. Co-expression of ADAM17 and glucose-regulated protein 78 (GRP78) was detected by immunofluorescence. mRNA expression of TGF-β1, Smad3, α-smooth muscle actin (α-SMA), collagen typeⅠalpha 1 chain (COL1A1), ADAM17, GRP78,inositol-requiring enzyme 1(IRE1)and activating transcription factor 6 (ATF6) RT-qPCR. Protein expressions of ADAM17, Smad3, TGF-β1, α-SMA, COL1A1, and GRP78 were measured by Western blot. Results Compared with the Control-1 group, rats in the Model-1 group exhibited decreased weight(P<0. 01), increased UPCR, and significantly higher levels of serum renal function markers ( P<0. 0001 ).Morphological examinations revealed pathological changes such as renal interstitial fibrosis. The expression of RF-related indicators TGF-β1 and Smad3 was upregulated (P<0. 001), accompanied by abnormal expression of ADAM17 and GRP78 (P<0. 01). Overexpression of ADAM17 (P<0. 0001) also led to a decrease in body weight, increased UPCR,and significant increases in serum renal function markers (P<0. 01), and pathological changes associated with RF in rats. The expression levels of RF-related indicators, including TGF-β1, Smad3, α-SMA, and COL1A1, were elevated to varying degrees (P<0. 05). Meanwhile, the expression of ERS-related factors GRP78, ATF6, and PERK was upregulated to varying degrees (P<0. 05). Conversely, knockdown of the ADAM17 alleviated the aberrant expression of the aforementioned indicators to varying degrees (P<0. 05). Conclusions ADAM17 significantly promotes fibrosis progression; its dysregulation of ERS may serve as a critical mechanism underlying this pro-fibrotic process.

Abstract: Objective To identify the effective components and mechanism of Bushen Zhuanggu Tang for the treatment of type 2 diabetic osteoporosis (T2DOP) using a comprehensive strategy including liquid chromatography /mass spectrometry (LC/ MS) and network pharmacology, combined with experimental verification. Methods The effective chemical components in Bushen Zhuanggu Tang were analyzed and identified by ultra-high-performance LCQ-Orbitrap-MS. Genes related to T2DOP and ferroptosis were retrieved from the database, and the intersection was obtained to identify potential targets of the active ingredients for the treatment of T2DOP by regulating the ferroptosis pathway. A T2DOP rat model was induced by intraperitoneal injection of streptozotocin combined with high-sugar and high-fat feeding, and the rats were then administered Bushen Zhuanggu Tang for 8 weeks. Body weight, blood glucose, and glycosylated hemoglobin were measured. Bone morphological changes were detected by Micro CT. Total iron ion, superoxide dismutase (SOD), and malondialdehyde (MDA) levels were detected using appropriate kits.Nuclear factor erythroid 2-related factor 2 ( Nrf2) / heme oxygenase-1 ( HO-1) signaling- and ferroptosis-related proteins in bone tissue were detected by Western blot. Results A total of 848 targets of 46 identified compounds were predicted in the blood compounds of Bushen Zhuanggu Tang. Comparing the GeneCards, Comparative Toxicogenomics, and FerrDb V2 databases finally indicated 81 potential targets of Bushen Zhuanggu Tang against T2DOP by regulating ferroptosis. Protein-protein interaction network analysis indicated that silent information regulator1, mammalian target of rapamycin, signal transducer and activator of transcription 3, Nrf2, SRC, epidermal growth factor receptor, interleukin ( IL)-6, mitogen-activated protein kinase-3, IL-1B, peroxisome proliferator-activated receptor γ, albumin, and other nodes may be important targets for Bushen Zhuanggu Tang in the treatment of T2DOP.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis identified 134 signaling pathways related to the intersection target genes. The main pathways were metabolic pathways, lipid and atherosclerosis, and ferroptosis signaling pathways in diabetic complications. The in vivo experiments showed that the body weight of rats in the Model group increased slowly compared with the Control group, while blood glucose and glycosylated hemoglobin levels increased, trabecular bone was destroyed, bone mineral density, bone volume fraction, and trabecular thickness and density decreased, the degree of trabecular separation increased, total iron ion and MDA levels in bone tissue increased, SOD activity decreased, and Nrf2, HO-1, and ferroptosis-related protein levels increased (P<0. 05).Compared with the Model group, weight gain increased, blood glucose and glycosylated hemoglobin levels decreased,destruction of the bone trabecula decreased, bone mineral density, bone volume fraction, and bone trabecula number and density increased, total iron ion and MDA levels in bone tissue decreased, while SOD activity increased, and Nrf2, HO-1, and ferroptosis-related protein levels decreased (P<0. 05). Conclusions Bushen Zhuanggu Tang has multi-component, multi-target, and multi-pathway characteristics for the treatment of T2DOP. The result of in vivo experiments show that Bushen Zhuanggu Tang can prevent and treat T2DOP by regulating the Nrf2 / HO-1 signaling pathway and inhibiting oxidative stress and ferroptosis.

Abstract: Objective To investigate the effects of ultrasound microbubble (UM)-mediated microRNA-545-3p (miR-545-3p) on the proliferation, migration, and invasion of thyroid cancer (TC) cells by regulating the E2F transcription factor 3 (E2F3) signaling pathway. Methods mRNA levels of miR-545-3p and E2F3 were detected in PTC cell lines (TPC-1, K1, TTA1) and TC tissues by quantitative reverse transcription-polymerase chain reaction. The relationship between miR-545-3p and E2F3 was detected by dual luciferase assay. Microbubbles were prepared using ultrasound technology, and TPC-1 cells were assigned to control, miR-negative control (NC), miR-545-3pmimics, UM+miR-545-3p-mimics, UM+miR-545-3p-mimics+pcDNA, and UM+miR-545-3p-mimics+E2F3 groups.The effects of ultrasound microbubble-mediated miR-545-3p on the proliferation, migration, invasion, and epithelialmesenchymal transition (EMT) of PTC cells by targeting E2F3 were detected. Expression levels of relevant proteins were detected by immunoblotting. Tumor transplant experiments were used to detect the effects of miR-545-3p on tumor growth and E2F3 expression. Results Expression levels of E2F3 mRNA were increased in TC tissues and TPC-1 cells, while expression levels of miR-545-3p decreased (P<0. 05). Online prediction of the Starbase database found that there was a targeted binding site between miR-545-3p and E2F3. The number of cell clones, scratchhealing rate, and number of invasive cells were lower in the miR-545-3p-mimics group compared with the miR-NC group, expression levels of E2F3, cyclin-dependent kinase (CDK) 1, matrix metalloproteinase (MMP)-2, MMP-9,N-Cadherin, and Vimentin were lower, while expression levels of miR-545-3p and E-Cadherin were higher ( P<0. 05). The number of cell clones, scratch-healing rate, and number of invasive cells were lower in the UM+miR-545-3p-mimics group compared with the miR-545-3p-mimics group, expression levels of E2F3, CDK1, MMP-2,MMP-9, N-Cadherin, and Vimentin were lower, while expression levels of miR-545-3p and E-Cadherin were higher(P<0. 05). Compared with the UM+miR-545-3p-mimics+pcDNA group, the number of cell clones, scratch-healing rate, and number of invasive cells were higher in the UM+miR-545-3p-mimics+E2F3 group, expression levels of E2F3, CDK1, MMP-2, MMP-9, N-Cadherin, and Vimentin were higher, and expression levels of miR-545-3p and E-Cadherin were lower (P<0. 05). Tumor volume and mass were smaller in the miR-545-3p-mimics group compared with the miR-NC group, while E2F3 expression levels were lower and miR-545-3p levels were higher (P<0. 05).Tumor volume and mass were smaller in the UM+miR-545-3p-mimics group compared with the miR-545-3p-mimics group, E2F3 expression levels were lower and miR-545-3p levels were higher ( P<0. 05). Conclusions UMmediated miR-545-3p inhibits the proliferation, migration, invasion, and EMT of PTC cells by targeting the E2F3 signaling pathway.

Abstract: Objective To investigate the effect of gastrodin (GAS) on inflammatory injury and the AMPdependent protein kinase / nuclear factor κB/ nucleotide-binding oligomerization domain-like receptor protein 3 (AMPK/ NF-κB/ NLRP3) pathway in rats with periodontitis. Methods A periodontitis rats model was constructed,and successfully modeled rats were randomly assigned into either the Model group, low, medium, and high-dose GAS (GAS-L, GAS-M, and GAS-H, respectively), or high-dose GAS+pathway inhibitor compound C group (GAS-H+CC), with 18 rats in each. Additionally, 18 healthy normal rats formed the Control group. The gingival index and the gingival bleeding index of rats were evaluated. Microscopic computed tomography was used to measure alveolar bone resorption, cemento-enamel junction-alveolar bone crest (CEJ-ABC) distance, and bone mineral density. Enzymelinked immunosorbent assay was used to detect levels of inflammatory factors in serum and periodontal tissues.Hematoxylin-eosin staining was performed to detect pathological damage in periodontal tissue. Tartrate-resistant acid phosphatase staining was performed to detect the number of osteoclasts in periodontal tissue. Immunohistochemical detection of the expression of bone metabolism-related proteins in periodontal tissues was performed, and Western blot was used to detect the expression of AMPK/ NF-κB/ NLRP3 pathway-related proteins. Results Compared with the Model group, the GAS-L, GAS-M, and GAS-H groups showed reduced periodontal tissue pathological damage, bone resorption cavities, inflammatory cell infiltration, gingival index, gingival bleeding index, CEJ-ABC distance, tumor necrosis factor (TNF)-α, interleukin (IL)-6, tissue damage score, osteoclast count, RANKL, p-NF-κB p65 / NF-κB p65, and NLRP3, while bone mineral density, osteoprotegerin (OPG), osteocalcin (OCN), and p-AMPK/ AMPK increased (P<0. 05,P<0. 01). The GAS-H+CC group showed more severe periodontal tissue pathological damage than the GAS-H group; the gingival index, gingival bleeding index, CEJ-ABC distance, TNF-α, IL-6, tissue damage score, osteoclast count, RANKL, p-NF-κB p65 / NF-κB p65, and NLRP3 increased, while bone mineral density,OPG, OCN, and p-AMPK/ AMPK decreased (P<0. 05,P<0. 01). Conclusions GAS improved inflammatory injury in periodontitis rats through regulating the AMPK/ NF-κB/ NLRP3 pathway.

Abstract: Objective To explore the effect and mechanism of quercetin (QCT) on peritoneal fibrosis (PF) in peritoneal dialysis (PD) rats based on the Wnt / β-catenin pathway. Methods PD rat model was constructed, and successfully modeled rats were stochastically classified into LiCl-N ( LiCl, 0 mg / kg), LiCl-L ( LiCl, 30 mg / kg)、LiCl-M (LiCl, 60 mg / kg), LiCl-H (LiCl, 90 mg / kg), PD, QCT-L (QCT, 17. 5 mg / kg), QCT-H (QCT, 35 mg /kg), and QCT-H+LiCl-M (QCT, 35 mg / kg+LiCl, 60 mg / kg) groups. Another 12 rats served as the Ctrl group.Rats in the Ctrl and PD groups were given an equal amount of 0. 9% sodium chloride solution at the same time and location by gavage once a day for 4 weeks. Peritoneal function was assessed using the peritoneal balance test. Changes in peritoneal tissue were observed by hematoxylin-eosin staining and peritoneal tissue fibrosis was detected by Masson staining. Levels of the inflammatory factors tumor necrosis factor ( TNF)-α, interleukin ( IL)-1β, and IL-6 were detected by enzyme-linked immunosorbent assay. α-smooth muscle actin (SMA), collagen type 1 (CoL-1), and Ecadherin protein levels were detected by immunohistochemistry, and Wnt3a, low-density lipoprotein receptor-related protein 5 (LRP5), lymphoid enhancer-binding factor 1 ( LEF-1), glycogen synthase kinase-(GSK)-3β, and β-catenin proteins were measured by Western blot. Results Compared with the LiCl-N group, rats in the LiCl-L groups showed no effects on serum creatinine ( Scr) and blood urea nitrogen ( BUN) levels, mass transfer of glucose (MTG), ultrafiltration volume (UF), Wnt3a, and β-catenin protein (P>0. 05), Rats in the LiCl-M groups showed same effects, while Wnt3a, β-catenin prote were increased (P<0. 05). Rats in the LiCl-H groups showed more Scr and BUN levels, mass transfer of glucose (MTG), and Wnt3a and β-catenin protein(P<0. 05), while UF were decreased (P<0. 05). Thus, 60 mg / kg was selected as the LiCl concentration. Rats in the PD group shed peritoneal mesothelial cells compared with the Ctrl group’ s number of inflammatory cells, relative area of collagen fiber deposition, peritoneal thickness, Scr and BUN levels, MTG, peritoneal tissue TNF-α, IL-1β, IL-6, α-SMA, CoL-1,Wnt3a, LRP5, LEF-1, and β-catenin were all increased (P<0. 05), while UF, E-cadherin, and GSK-3β proteins decreased (P<0. 05). Compared with the PD group, rats in the QCT-L and QCT-H groups showed less mesothelial cell shedding, number of inflammatory cells, relative area of collagen fiber deposition, peritoneal thickness, SCr,BUN, MTG, peritoneal tissue TNF-α, IL-1β, and IL-6 levels, and α-SMA, CoL-1, Wnt3a, LRP5, LEF-1, and β-catenin proteins were decreased ( P<0. 05), while UF, E-cadherin, and GSK-3β were increased ( P<0. 05).Compared with the QCT-H group, rats in the QCT-H + LiCl-M group showed increased peritoneal tissue damage,increased number of inflammatory cells, relative area of collagen fiber deposition, peritoneal thickness, Scr and BUN levels, MTG, peritoneal tissue TNF-α, IL-1β, and IL-6 levels, and α-SMA, CoL-1, Wnt3a, LRP5, LEF-1, β-catenin proteins (P<0. 05), and decreased UF, E-cadherin, and GSK-3β (P<0. 05). Conclusions QCT alleviates PF in PD rats by inhibiting the inflammatory response and peritoneal mesothelial cell mesenchymal transition process, possibly acting via inhibition of the Wnt / β-catenin signaling pathway.

Abstract:Most laboratory animal centers in colleges and universities have adopted or developed management systems for laboratory animals; however, these systems vary significantly in terms of functional modules, management refinement level, and operational efficiency. Based on an analysis of the current status of laboratory animal management systems in colleges and universities, combined with the practice experience of our institution, this study develops a refined, full-process information management system centered on cage, which significantly improves the efficiency and transparency of laboratory animal management, enhances the level of refined management and quality control, and promotes resource integration and open sharing. This system provides a new approach for the refined management of laboratory animals in colleges and universities.

Abstract:Synaptic pruning, as a core process in neural development and functional maintenance in the mammalian central nervous, relies on precise molecular and cellular regulation and is crucial for constructing efficient and stable neural networks. Dysregulation of synaptic-pruning mechanisms disrupts the dynamic equilibrium of neural connections, closely associated with the onset and progression of various central nervous system diseases. The mechanisms underlying glial cell-mediated synaptic elimination are not yet fully understood; however, multiple signals promoting or inhibiting synaptic clearance are known to be involved in collectively regulating this complex process. Indepth analysis of the cellular and molecular mechanisms governing synaptic formation and elimination can not only reveal the underlying pathogenesis of central nervous system diseases, but may also provide a theoretical basis for their early diagnosis and intervention, and for the development of innovative therapies. Further exploration of synapticpruning mechanisms has the potential to result in critical breakthroughs in tackling neurological disorders.

Abstract:Cardiovascular disease is currently the disease with the highest incidence and mortality rate globally, but pathogenesis research and drug development for this condition still face significant challenges, partly because of the limitations of traditional research models. Two-dimensional ( 2D) cell cultures make it difficult to simulate the microenvironment in vivo, while animal models have species differences that cannot fully reproduce the physiological and pathological characteristics of the human cardiovascular system. With the continuous breakthrough of stem cell technology, organoid model research has developed rapidly, providing innovative tools for cardiovascular mechanism research and drug development. Cardiovascular organoids are microtissue models constructed with stem cells under three-dimensional (3D) culture conditions in vitro; they can highly simulate the structure, composition,and function of the human heart and blood vessels, significantly improving the physiological relevance of research. This paper systematically reviews their construction strategies and application progress, discussing current limitations and future directions.

Abstract:Alzheimer’ s disease (AD) is a neurodegenerative disorder primarily characterized by cognitive impairment and executive function deficits. As a crucial innate immune pathway, abnormal activation of the cyclic GMP-AMP synthase-stimulator of interferon gene (cGAS-STING) signaling pathway in AD has attracted considerable attention. This review discusses the involvement of the cGAS-STING signaling pathway in AD via mechanisms including β-amyloid deposition and abnormal tau protein aggregation, oxidative stress and mitochondrial dysfunction,neuroinflammation and glial cell abnormalities, synaptic dysfunction, vascular dysfunction, gut microbiota dysbiosis, and autophagy dysfunction, as well as considering relevant research on potential therapeutic interventions in the cGAS-STING signaling pathway for the treatment of AD. This review aims to reveal the role of abnormal activation of the cGAS-STING signaling pathway as a cause of AD pathogenesis and as a potential interventional target for the treatment of AD.

Abstract:In recent years, declining male fertility and increasing incidence of testis-related diseases have become increasingly prominent, with environmental toxins, genetic factors, and lifestyle being the main contributing factors. Assessment of male reproductive toxicity has emerged as a critical link in environmental safety and drug development processes. Traditional animal experiments suffer from several limitations, including long experimental cycles, ethical controversies, interspecies differences, and the inability to simulate the testicular microenvironment.With the implementation of the 3R principle (reduction, replacement, and refinement), the construction of in vitro models with high physiological relevance has becomeinevitable. 3D cell culture can mimic the in vivo environment and provide a suitable 3D niche for testicular cells. This review summarizes the research progress into 3D cell culture for the development of testicular models. The composition of the testicular germ cell niche and its significance for toxicity assessment are detailed, and the characteristics and efficacy of five 3D culture systems including decellularized testicular scaffolds and hydrogel scaffolds, from the perspectives of their technical principles and applications are analyzed. Additionally, existing limitations of current models are addressed, proposing that future research should focus on standardization, multicell co-culture, and high-throughput screening. These efforts are intended to promote the application of 3D testicular models in reproductive toxicity research, providing support for environmental safety assessment and drug development.

Abstract:The global incidence of diabetes mellitus has recently shown a steep rise. Diabetic kidney disease (DKD), as one of the most prevalent microvascular complications, has accordingly become the primary cause of endstage renal disease. There is an urgent need to identify safe, reliable, cost-effective, and low-side-effect medications for early interventions for DKD to delay disease progression. Pathological studies have revealed that the adenylate monophosphate-activated protein kinase ( AMPK) signaling pathway plays a central role in DKD pathogenesis,offering a novel therapeutic direction by regulating inflammatory responses, oxidative stress, autophagy, apoptosis,lipid metabolism, ferroptosis, and other biological processes. Traditional Chinese medicine (TCM) has been shown to effectively alleviate edema, proteinuria, and declining renal function, while significantly improving patients’ quality of life—a critical consideration for early DKD intervention. Notably, terpenoids, flavonoids, phenolic compounds, alkaloids, and several TCM formulations ( e. g. , Shuilu Dihuang capsules, Angelica Sinensis Blood Tonic soup,Tangshenning formula, Jinlida granules, and Colquhounia Root tablets) have exhibited significant AMPK pathwaytargeting effects. This review comprehensively synthesizes recent domestic and international advancements in TCMmediated regulation of the AMPK signaling pathway in DKD, aiming to provide a reference for understanding the mechanisms and clinical efficacy of TCM in treating DKD.

Abstract:Physiological telemetry technology enables the real-time monitoring of physiological parameters in freely moving organisms using wireless transmission, thus overcoming the spatial limitations of traditional detection method. This provides reliable physiological data in fields such as cardiovascular pharmacology, neuroscience, and metabolic research. This paper systematically reviews the technical principles and current status of implantable and non-implantable systems, summarizing core technological advancements such as sensor miniaturization, optimized wireless energy transmission, and the application of intelligent algorithms. The review further analyzes the adaptability of different experimental animal models and their application value in fields such as cardiovascular medicine and neuroscience. Future developments will focus on continuing breakthroughs in flexible electronics, intelligent algorithms, and standardized operations to achieve more precise and universal physiological monitoring. By integrating artificial intelligence and 3D printing technology, telemetry technology will be widely applicable in both basic research and clinical translation.