Abstract: Objective This study investigated the effects of cigarette smoke extract ( CSE)-induced macrophage-conditioned medium on airway epithelial cells?? inflammatory responses, and explored the underlying mechanisms involved. Methods Phorbol myristate acetate-induced differentiated monocytes and macrophages (THP-1) were cultured with 10% CSE for 24 h; the conditioned medium was collected to induce inflammatory responses in human bronchial epithelial cells (BEAS-2B). The effects of THP-1-conditioned medium on BEAS-2B cells viability were detected using the CCK-8 assay. The effects of conditioned medium on mRNA expression levels of inflammationrelated factors in BEAS-2B cells were analyzed by RT-qPCR, and inflammatory cytokine secretion was measured with enzyme-linked immunosorbent assay. The effects of conditioned medium on BEAS-2B cells morphology were observed with immunofluorescence staining. The expression of proteins related to the mTOR/ p70S6K pathway was examined by Western blot. Results BEAS-2B cells viability after treatment with 6. 25%, 12. 5%, or 25% conditioned medium showed no significant changes after 6, 12, 24, or 48 h, while 50%, 75%, and 100% induced significant reductions in cell viability (P<0. 05, P<0. 01). Conditioned medium at 6. 25%, 12. 5%, and 25% significantly increased the mRNA expression and secretion of inflammatory factors interleukin ( IL)-1β, IL-6, IL-8, and TNF-α in BEAS-2B cells(P<0. 05, P<0. 01), but had no significant effect on morphology. Levels of p-mTOR, p-p70S6K, p-GSK3β (Tyr216), and p-NF-κB p65 were significantly increased in conditioned medium-treated cells (P<0. 05, P<0. 01);the mTORC1 inhibitor rapamycin significantly reduced inflammatory factor mRNA expression in BEAS-2B cells induced by 25% conditioned medium. Conclusions CSE-induced macrophage-conditioned medium significantly induced airway epithelial cell inflammatory responses through activating the mTORC1 / p70S6K pathway.

Abstract: Objective This study explored the effects of zinc finger and BTB domain protein 20 (ZBTB20) on rat hepatic stellate cell (HSC-T6) proliferation and migration, providing experimental evidence for hepatic fibrosis (HF) treatment. Methods ZBTB20 expression in liver tissues of normal person and HF patients were analyzed by the GEO database. Ten SD rats were randomly divided into Sham operation and common bile duct ligation (BDL) groups to establish a cholestatic HF model. Western blot assay was used to detect the expression levels of ZBTB20 in rat liver tissues. Western blot assay was used to detect the expression levels of ZBTB20 and fibrosis markers in HSCT6 cells after 48 hours of transforming growth factor TGF-β1 stimulation. RT-qPCR and Western blot assays were used to detect the expression levels of the transfection efficiency of ZBTB20 small interfering RNA ( si-ZBTB20) and overexpression plasmid (pcDNA3. 1-ZBTB20), as well as the expression levels of fibrosis markers in HSC-T6 cells in the si-ZBTB20 and pcDNA3. 1-ZBTB20 groups. The effects of si-ZBTB20 / pcDNA3. 1-ZBTB20 on HSC-T6 cells proliferation, colony formation, and migration were detected through CCK-8, colony formation, and transwell assays. Results ZBTB20 was significantly highly expressed in the liver tissues of HF patients. ZBTB20 expression levels were significantly up-regulated in the BDL group compared with the Sham group (P<0. 05). α-SMA, Col1A1, and ZBTB20 proteins were significantly up-regulated in HSC-T6 cells in the TGF-β1 group compared with the control group (P<0. 05). RT-qPCR and Western blot assay showed that ZBTB20, α-SMA, Col1A1, and CTGF mRNA expression was down-regulated in the si-ZBTB20-transfected group compared with the si-NC-transfected group, and ZBTB20,α-SMA, and Col1A1 protein expression was decreased (P<0. 05); the above indicators in the pcDNA3. 1-ZBTB20- transfected group showed opposite trends compared with the pcDNA3. 1-transfected group. CCK-8, colony formation,and Transwell assays showed that cell proliferation in the si-ZBTB20-transfected group decreased significantly after 72 and 96 hours of transfection, the number of cell colonies decreased significantly, and the number of cells passing through the basement membrane of the Transwell chamber decreased significantly compared with the si-NC-transfected group (P<0. 05); the above indicators in the pcDNA3. 1-ZBTB20-transfected group showed opposite trends compared with the pcDNA3. 1-transfected group. Conclusions ZBTB20 was significantly highly expressed in the liver tissues of patients with HF, and its expression in BDL rat liver tissues increased. ZBTB20 knockdown inhibited HSC-T6 cells proliferation and migration, while its overexpression promoted proliferation and migration.

Abstract: Objective To assess the effects of long-term feeding of transgenic cry1Ab / cry2Aj and G10evoepsps insect-resistant and herbicide-tolerant maize on bone mineral content ( BMC) and density ( BMD) in two generations of Macaca fascicularis. Methods A total of 24 parental ( F0) cynomolgus monkeys were divided into 70% genetically modified corn(GM; n= 8), 70% non-genetically modified corn (Non-GM; n= 7), and normal feed control groups (Normal; n= 9). The offspring (F1) generation included 46 Macaca fascicularis, comprising 16 in the GM group, 13 in the Non-GM group, and 17 in the Normal group, with age ranges spanning 1 ~ 6 years. Lunar densitometers were used to scan and analyze BMC and BMD in the head, arms, legs, pelvis, spine, and whole body of animals in each group. Results No significant BMC and BMD differences ( P>0. 05) were observed between groups in the head, arms, legs, pelvis, spine, or whole body at three age stages (1 ~ 2, 3 ~ 4, and 5 ~ 6 years) in both F0 and F1 generations. Conclusions Long-term feeding with insect-resistant and herbicide-tolerant transgenic maize did not significantly affect BMC and BMD in either parental or offspring generations of Macaca fascicularis. This study provides reference data for evaluating the long-term safety of consuming this genetically modified maize.

Abstract: Objective This study systematically evaluated the efficacy and mechanism of action of Zhenwu Tang in treating diabetic kidney disease (DKD) using animal models. Methods Chinese and English databases (PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang Database, Chinese Biomedical Literature Service System, and VIP Information ) were searched from inception until June 2025. Experimental studies investigating Zhenwu Tang intervention in DKD animal models were included. The systematic review centre for laboratory animal experimentation tool was used to assess the quality, the GRADE-CERQual framework was applied to evaluate the certainty of evidence, and RevMan 5. 4 was used for statistical analysis. Results Thirteen studies were included, involving db / db mice and SD rat models. The primary modeling Methods induded streptozotocin ( STZ) injection or STZ combined with a high sugar and fat diet. Spleen-kidney yang deficiency syndrome was induced using a decoction of raw Rheum palmatum combined with hydrocortisone. Compared with model groups, Zhenwu Tang intervention significantly reduced key indicators, including serum creatinine, blood urea nitrogen, fasting plasma glucose, and 24-hour urinary protein excretion in DKD animals, though some studies did not observe a significant improvement in fasting plasma glucose with Zhenwu Tang intervention; it also ameliorated glomerulosclerosis, tubular injury, and renal fibrosis. The renal protective effects of Zhenwu Tang involve a multi-target network centered on crosstalk between suppressed nuclear factor-κB-mediated inflammation and activation of Nrf2-driven anti-oxidation,interconnected through reactive oxygen species modulation, leading to downstream effects such as key podocyte protection (upregulation of the key podocyte protein nephrin) and anti-fibrosis. The certainty of evidence was graded as “High” for biochemical parameters, urinary protein, and histopathological findings; “Moderate” for inflammation and oxidative stress markers; and “Low” for podocyte protein expression. Conclusions Zhenwu Tang exerts a renal protective effect in DKD animal models through a multi-target network centered on “ anti-inflammatory-antioxidant”crosstalk, which is dose- and time-dependent. Future more rigorous and high-quality basic studies are needed to verify these findings and further explore the optimal dose window and mechanism of action.

Abstract: Objective The collateral circulation represents an endogenous anastomotic route that compensates for the stenosis or occlusion of intracranial arteries. It supplies collateral blood flow, thereby sustaining the survival of the ischemic penumbra. This study compared the labeling of cerebral collateral arteries in mice in vitro and in vivo, using different experimental method. Methods Primary ( Willis circle ) and secondary (leptomeningeal) collateral arteries in mice were labeled in vitro using latex, DiI and ink-gelatin perfusion techniques, respectively. The diameter of the collateral arteries was then measured. Specifically, latex perfusion was employed to compare the number of leptomeningeal collaterals(LMCs) between different mouse strains. Stable in vivo observation of LMCs was achieved by constructing a closed cranial window. A mouse model of distal middle cerebral artery occlusion stroke was established via electrocoagulation, followed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Additionally, LMC diameter was continuously monitored using a fluorescence stereomicroscope, and blood flow in the peri-infarct area was detected by laser speckle contrast imaging. Results Latex and DiI perfusion both labeled the collateral arteries, but not veins, with no significant difference in vessel diameter measured by the two method ( P>0. 05 ). DiI perfusion often exhibited suboptimal perfusion efficiency. The number of LMCs was significantly higher in C57BL / 6N compared with BALB/ c mice (P<0. 000 1). The clarity and imaging quality of the cranial window were not significantly affected during the 3-week closed cranial window period. TTC staining confirmed stable construction of the mouse stroke model. LMC diameter increased and the blood flow in the peri-infarct area recovered after ischemic stroke (P<0. 05), with a strong positive correlation between them (P<0. 05). Conclusions Latex perfusion can effectively label primary and secondary collateral arteries in mice in vitro. Fluorescence stereomicroscopy and laser speckle contrast imaging can be used for continuous observations of LMCs in vivo.Remodeling of collateral vessels after ischemic stroke may promote the recovery of blood flow in the penumbra.

Abstract: Objective To investigate the effects of urothelial carcinoma antigen 1 long non-coding RNA(LncRNA UCA1) on the proliferation, apoptosis, and radiosensitivity of bladder cancer cells by regulating miR-520a-5p. Methods LncRNA UCA1 and miR-520a-5p were detected in normal bladder epithelial (HCV-29) and human bladder cancer (SW780, HT1376, BIU87 and T24) cell lines by quantitative reverse transcription-polymerase chain reaction. T24 cells were treated with different radiation doses (0~8 Gy) and cell proliferation activity was detected.T24 cells were assigned to Control, sh-NC, sh-UCA1, sh-UCA1 + anti-miR-NC, and sh-UCA1 + anti-miR-520a-5p groups. Cells in each group were treated with a radiation dose of 2 Gy, and cell proliferation, apoptosis, and cyclin D1, Ki-67, Bax, and Caspase-3 protein expression levels were detected. The targeting relationship between LncRNA UCA1 and miR-520a-5p was verified by dual-luciferase reporter gene, fluorescence in situ hybridization, and RNA immunoprecipitation assays. A nude mouse tumor xenotransplantation model was established, and the effect of LncRNA UCA1 on the radiosensitivity of bladder cancer was verified. Results LncRNA UCA1 was increased and miR-520a-5p was decreased in bladder cancer cell lines. Treating T24 cells with different radiation doses (0~8 Gy) reduced cell proliferation activity in the sh-UCA1 group compared with the sh-NC group, in line with increasing radiation dose (P<0. 05). T24 cells and a radiation dose of 2 Gy were therefore selected for subsequent experiments. Knockdown of LncRNA UCA1 in T24 cells inhibited cell proliferation, promoted cell apoptosis, increased radiosensitivity, down-regulated cyclin D1 and Ki-67, and upregulated Bax and Caspase-3 (P<0. 05). miR-520a-5p antagonist weakened the impacts of LncRNA UCA1 knockdown on the proliferation, apoptosis, and radiosensitivity of bladder cancer cells (P<0. 05). LncRNA UCA1 targeted the negative regulation of miR-520a-5p. Knockdown of LncRNA UCA1 combined with radiotherapy inhibited the growth of bladder cancer tumor transplants and increased their radiosensitivity (P<0. 05). Conclusions LncRNA UCA1 is up-regulated in bladder cancer cells. Knockdown of LncRNA UCA1 can inhibit cell proliferation, induce apoptosis, and enhance the radiosensitivity of bladder cancer cells by up-regulating miR-520a-5p.

Abstract: Objective To explore the possible mechanism of spindle and centromere associated complex subunit 3 ( SKA3 ) in regulating the malignant biology of oral squamous cell carcinoma ( OSCC ). Methods Expression levels of SKA3 in oral squamous cell carcinoma (n= 25) and adjacent tissues (n= 15) were detected by immunohistochemistry. The proliferation of CAL27 cells was assessed using a cell counting kit-8 assay,while migration and invasion were evaluated via scratch wound and Transwell assays, respectively. Expression levels of SKA3, protein tyrosine phosphatase non-receptor type 14 (Ptpn14), and Yes-associated protein-1 (YAP1) were detected by Western blot, and p53-binding protein 1 ( 53BP1) and the histone variant γH2AX were detected by immunofluorescence. The cell cycle and apoptosis rate were measured by flow cytometry. Results SKA3 expression levels were higher in clinical OSCC than in normal oral mucosal tissue samples(P<0. 01). Silencing SKA3 increased Ptpn14(P<0. 01) and inhibited YAP1 protein in cell experiments(P<0. 05), while overexpression of SKA3 had the opposite effects(P<0. 01). Silencing SKA3 also reduced the proliferation, migration, and invasion of OSCC cells(P<0. 01), blocked the G2/ M phase of the cell cycle(P<0. 01), and promoted DNA damage and cell apoptosis(P<0. 01) in CAL27 cells, while the YAP agonist XMU-MP-1 reversed the effect of silencing SKA3 ( P<0. 05, P<0. 01). Conclusions Silencing SKA3 inhibits YAP1 protein expression by activating the Hippo / YAP signaling pathway, thereby inhibiting the proliferation, migration, and invasion of OSCC cells, and promoting DNA damage and cell apoptosis.

Abstract:Premature ovarian insufficiency (POI) is a condition that severely impacts female physical and mental health; it is a primary cause of female infertility, with its incidence rate continuously rising in recent years. Although hormone replacement therapy has achieved good efficacy in clinical practice, its long-term use can lead to many adverse reactions. Finding safe and effective POI treatments has become an urgent issue. Traditional Chinese medicine can regulate phosphatidylinositol 3-kinase / protein kinase B (PI3K/ Akt), Wnt family secreted glycoprotein /β-catenin (Wnt / β-catenin), transforming growth factor-β (TGF-β) / Smads, and nuclear factor erythroid 2-related factor ( Nrf2) signaling, along with other related signaling pathways. It can also inhibit follicular granulosa cell apoptosis, reduce inflammatory responses and oxidative stress damage, regulate hormone levels, improve ovarian function, and delay ovarian aging. Traditional Chinese medicine presents advantages in POI treatment, with significant clinical effects and high safety. Through exploring research into the signaling pathways in POI, this paper reviews and analyzes literature on the active components of traditional Chinese medicine and their use in POI treatment by modulating related signaling pathways, aiming to offer guidance for future research.

Abstract:Alzheimer’ s disease (AD) is a neurodegenerative disease characterized by cognitive decline,with neuroinflammation being a key driver. As an important transcription factor, cyclic AMP response element binding protein (CREB) is involved in neuroinflammation, synaptic plasticity, and neuronal survival through regulation of multiple signaling pathways. This article reviews the biological functions of CREB, systematically explains the regulatory mechanisms involved in CREB-mediated signaling pathways in AD related to neuroinflammation, including cAMP / PKA, MAPK/ ERK, and PI3K/ AKT, and summarizes the research progress on CREB agonists of small molecule drugs, natural compounds, and cAMP analogues. Precise CREB regulation may provide a new strategy for AD treatment and provide a theoretical basis for identifying new AD therapeutic targets and drugs. This article reviews the research progress of CREB-mediated neuroinflammation in AD.

Abstract:Chronic kidney disease ( CKD) is a globally prevalent chronic progressive disease that often coexists with atherosclerosis (AS); these conditions mutually reinforce each other, accelerating the occurrence of cardiovascular events and significantly impacting prognosis. While traditional risk factors can partly explain disease progression, CKD-specific factors such as uremic toxin accumulation, calcium-phosphorus metabolism disorders, the renin-angiotensin system, chronic inflammation, and oxidative stress also play important roles in AS progression. The RhoA/ ROCK pathway plays a crucial role in pathological processes such as vascular smooth muscle cell contraction,endothelial dysfunction, inflammatory responses, and oxidative stress, making it a key molecular mechanism in CKD and AS development. As a member of the small GTPase family, RhoA can activate downstream ROCK1 / ROCK2,leading to actin cytoskeletal remodeling and vascular tone regulation. In CKD, factors such as uremic toxin accumulation, elevated oxidative stress levels, and activation of the renin-angiotensin system can upregulate RhoA/ROCK signaling, further accelerating endothelial damage and AS plaque formation. Traditional Chinese medicine (TCM) and its active components have shown potential in interventional studies targeting this pathway. Various drugs, such as hirudin, hydroxy crocin, and rhodiola glycoside, can downregulate RhoA and ROCK expression or activity, improve endothelial function, and inhibit inflammation and oxidative stress, slowing the progression of CKD combined with AS. This study systematically evaluated the role of the RhoA/ ROCK pathway in CKD combined with AS and the progress of TCM interventions to help reveal the molecular disease mechanisms and provide new insights and strategies for clinical prevention and treatment.

Abstract:Post-stroke cognitive impairment ( PSCI) is one of the most common complications of stroke,seriously compromising quality of life and rehabilitation; it has become an urgent challenge in the field of stroke management. microRNAs ( miRNAs) play pivotal regulatory roles in the pathological processes underlying PSCI,including inflammation, apoptosis, oxidative stress, autophagy, and synaptic plasticity. Traditional Chinese medicine (TCM) has shown unique advantages and prospects in PSCI treatment, exerting its effects through overall regulation,multi-component synergy, and multi-target modulation of miRNA-mediated mechanisms. This review systematically summarizes the mechanism of miRNA in improving cognitive impairment and repair after stroke, and highlights recent progress on TCM-based interventions targeting miRNAs in PSCI. The findings provide a reference for miRNAs as potential biomarkers and therapeutic targets, and offer a theoretical basis for advancing the modernization of TCM in PSCI management.

Abstract:Drug screening is crucial in drug development; animal models serve as an essential foundation for disease research and screening. The selection of suitable models can elucidate disease mechanisms, reduce research costs, and expedite research. Caenorhabditis elegans ( C. elegans ), a multicellular eukaryote, has become an invaluable tool in life sciences research because of its minimal ethical concerns, short life cycle, low cost, ease of experimental manipulation, fully sequenced genome, and significant homology with the human genome. This article synthesizes recent literature to review the progress in drug screening using C. elegans, including its application in aging, age-related neurodegenerative disease, anti-tumor, and anti-infection models; additionally, effective drugs screened with these models are summarized, aiming to provide theoretical references and foundations for future research.

Abstract:Schizophrenia ( SCZ ) is a severe mental disorder characterized by complex pathological mechanisms and high treatment resistance; it currently ranks among the diseases with the heaviest socioeconomic burden worldwide. Current antipsychotic medications are limited by factors such as insufficient response rates, limited cognitive improvement, and significant metabolic side effects. The ketogenic diet (KD), a metabolic intervention strategy characterized by high fat, low carbohydrate, and moderate protein intake. Although current clinical trials of KD for SCZ have small sample sizes, emerging data suggest that KD can effectively alleviate psychiatric symptoms,cognitive dysfunction, and abnormal metabolic parameters in some patients through multiple pathways, including modulating cerebral energy metabolism, reducing oxidative stress, and balancing neurotransmitter systems, showing significant clinical translational value. This article systematically reviews the research progress of the KD as an adjunctive treatment for SCZ, discussing mechanisms of action, preclinical evidence, clinical exploration, current challenges, and future research directions.