Aging is a complex biological process that involves multiple organs and dimensions. Owing to its high resolution, single-cell (nucleus) transcriptomics enables in-depth exploration of the cellular and molecular mechanis ms underlying aging and the identification of aging biomarkers and therapeutic targets for relevant diseases. This review discusses the advantages of single-cell (nucleus) transcriptomic sequencing technologies in the study of aging biomarkers, summarizes their applications in elucidating anti-aging strategies and screening anti-aging compounds, and highlights their value in uncovering the pathological mechanisms of aging-related diseases.

Single-cell omics technologies are advancing pituitary research, achieving breakthroughs in deciphering pituitary cell heterogeneity, developmental processes, physiological functions, and mechanisms of related diseases. The paper systematically reviews recent advances in the use of single-cell omics technologies in pituitary biology, highlighting key findings regarding the construction of a pituitary cell atlas, elucidation of developmental regulatory mechanisms, revelation of adaptive alterations under pathological conditions, and the exploration of heterogeneity in pituitary neuroendocrine tumors. It further discusses the prospects and challenges of single-cell omics technologies applications in pituitary research, providing a theoretical basis for comprehensively unraveling the cellular and molecular mechanisms underlying pituitary physiology and pathology.

Objective: To evaluate the ability of human cell free fat extract (CEFFE) t o alleviate senescence in 3T3-L1 cells and to determine its effects on cellular proliferation and adipogenic differentiation. Methods: An in vitro senescence model was generated by exposing 3T3-L1 cells to doxorubicin, followed by treatment with or without CEFFE intervention. Cellular senescence was assessed by senescence-associated β-galactosidase (SA-β-gal) staining. The mRNA expression levels of senescence-related and adipogenic differentiation-related genes were detected by RT-qPCR. The protein expression levels of senescence marker proteins (p21 and γ-H2AX) were determined by Western blotting. Cell proliferation ability was evaluated by EdU staining. Results: CEFFE treatment significantly reduced the proportion of SA-β-gal-positive senescent cells and increased the percentage of EdU-positive proliferating cells. The expression of senescence-associated genes and senescence marker proteins was markedly downregulated, whereas adipogenic differentiation-related genes were significantly upregulated. Conclusion: CEFFE intervention effectively attenuates senescence in 3T3-L1 cells while promoting both proliferative activity and adipogenic differentiation, suggesting its therapeutic promise in modulating cellular aging and enhancing regenerative capacity.

Objective: To delineate depot-specific features of senescence in human subcutaneous mature adipocytes and to compare how adipocytes from different anatomical sites respond to pro-senescent stimuli. Methods: Publicly available transcriptomic datasets were firstly analyzed to compare the gene expression patterns between isolated adipocytes and whole adipose tissue. Subcutaneous fat was collected from human chest, abdomen, and thigh, and mature adipocytes were isolated. Senescence in vivo was evaluated by senescence-associated β-galactosidase (SA- β-gal) staining and quantitative PCR analysis of senescence-associated genes including cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin D1 (CCND1), and tumor protein p53 (TP53). For in vitro modeling of obesityrelevant stress, adipocytes were cultured in medium supplemented with 10% fetal bovine serum for 4–7 d; cellcycle re-entry (EdU incorporation), senescence markers, and senescence-associated secretory phenotype (SASP) factors interleukin-1β (IL-1β) and matrix metalloproteinase 14 (MMP14) were then measured. Results: Senescenceassociated markers were more prominent in mature adipocytes than in whole adipose tissue. Among non-obese individuals, thigh adipocytes displayed the earliest senescence features, with higher SA-β-gal activity and increased expression of CDKN1A and CCND1. After serum exposure, chest adipocytes exhibited the strongest response, characterized by a higher frequency of cell-cycle re-entry and greater induction of senescence markers and inflammatory/pro-fibrotic factors, including IL-1β and MMP14, than abdominal or thigh adipocytes. Conclusions: Subcutaneous adipocyte senescence displays clear depot specificity. Thigh adipocytes show earlier senescence features, whereas chest adipocytes are more responsive to obesity-related stimulation. This spatiotemporal heterogeneity may play an important role in metabolicregulation and development of obesity-associated diseases, incl uding breast cancer.

Objective: To investigate the effects of daidzein (DZ) on chondrocyte apoptosis and the silent information regulator 1 (SIRT1)/AMP-activated protein kinase (AMPK) pathway in knee osteoarthritis (KOA) rats. Methods: SD rats were divided into six groups: control group, model group, low-dose DZ group (5 mg/kg), medium-dose DZ group (10 mg/kg), DZ high-dose group (20 mg/kg) and positive control group (0.017 g/kg Viartril-S). An adjuvant-induced KOA rat model was established in all groups except the control group. After successful modeling, the rats were treated with the corresponding medications. H-E staining was performed to measure the pathological changes of cartilage tissue in KOA rats; ELISA was adopted to observe the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in cartilage tissue of KOA rats. TUNEL staining was applied to detect chondrocyte apoptosis; immunohistochemistry was used to determine the expressions of SIRT1 and p-AMPK in cartilage. CCK-8 assay was utilized to assess the proliferation of chondrocytes; flow cytometry was employed to measure apoptosis of chondrocyte. Western blotting analysis was conducted to evaluate the expression of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), SIRT1 and p-AMPK in chondrocytes. Results: Low, medium and high doses of DZ or Viartril-S alleviated the pathological injury of rat cartilage tissue, reduced the percentage of TUNEL-positive cells, the levels of IL-1β, TNF-α, apoptosis rate, and Bax protein expression, and improved the rate of SIRT1 and p-AMPK positive cells, the proliferation ability of chondrocytes, and the protein expression of Bcl-2, SIRT1 and p-AMPK. Conclusion: DZ can inhibit the apoptosis of chondrocytes in KOA rats, and the mechanism may be related to the activation of the SIRT1/AMPK pathway.

Objective: To investigate the therapeutic effects and mechanisms of hyperoside on myocardial damage in spontaneously hypertensive rats (SHR). Methods: SHR rats were randomly divided into SHR group, hyperoside group, positive control (losartan) group, recombinant high mobility group box 1 (rHMGB1) group, and hyperoside +rHMGB1 group. Additionally, Wistar Kyoto rats served as control group. After the intervention, blood pressure and cardiac function indicators were measured in rats. Serum samples were collected to detect the levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in the supernatant. Tissue slices were prepared to assess pathological and fibrotic changes in cardiac tissue. The mRNA expression of interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), and HMGB1/toll-like receptor 4 (TLR4) signaling pathway-related proteins in tissues were detected. Results: Compared with the control group, the SHR group showed lower left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), while demonstrating higher systolic and diastolic blood pressure, ANP and BNP levels, fibrosis area ratio, left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic dimension (LVESD), TNF-α and IL-1β mRNA expression, and HMGB1 and TLR4 protein expression. Compared with the SHR group, the hyperoside group and the positive control group had higher LVEF and LVFS, while indicating lower systolic and diastolic blood pressure, ANP and BNP levels, fibrosis area ratio, LVEDD, LVESD, TNF-α, IL-1β mRNA expression,and HMGB1 and TLR4 protein expression. The rHMGB1 group showed lower LVEF and LVFS, and higher systolic and diastolic blood pressure, ANP and BNP levels, fibrosis area ratio, LVEDD, LVESD, TNF-α and IL-1β mRNA expression, and HMGB1 and TLR4 protein expression. Compared with the rHMGB1 group, the hyperoside+rHMGB1 group exhibited higher LVEF and LVFS, and lower systolic and diastolic blood pressure, ANP and BNP levels, fibrosis area ratio, LVEDD, LVESD, TNF-α and IL-1β mRNA expression, and HMGB1 and TLR4 proteins expression were decreased. Compared with the hyperoside group, the hyperoside+rHMGB1 group displayed lower LVEF and LVFS, and higher systolic and diastolic blood pressure, ANP and BNP levels, fibrosis area ratio, LVEDD, LVESD, TNF-α and IL-1β mRNA expression, and HMGB1 and TLR4 protein expression. Conclusion: Hyperoside can alleviate myocardial damage in SHR rats by suppressing the HMGB 1/TLR4 signaling pathway.

Objective: To investigate the effect of andrographolide (AP) on vascular endothelial inflammation and high mobility group box 1 (HMGB1)/receptor of advanced glycation end product (RAGE) signaling pathway in rats with thrombosis angiitis obliterans (TAO). Methods: A TAO rat model was established and the successfully modeled rats were randomly divided into a model group (TAO group), low-dose and high-dose andrographolide groups (AP-L and AP-H groups), and high-dose andrographolide+pathway activator group (AP-H+rHMGB1 group), with 12 rats in each group. Another 12 healthy rats served as the control group (Control group). The TAO lesions of rats in each group were graded and scored. Blood viscometer was used to measure whole blood and plasma viscosity. ELISA was utilized to detect levels of thrombotic factors. H-E staining was applied to observe the pathological morphology of femoral artery tissue. Western blotting was employed to detect the protein expression related to vascular endothelial inflammation and HMGB1/RAGE signaling pathway. Results: Compared with the Control group, the TAO group exhibited more severe shedding of endothelial cells in the femoral artery tissue, with a large amount of thrombus formation. The whole blood viscosity, plasma viscosity, thromboxane B2 (TXB2) levels, and expressions of IL-6, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), HMGB1, RAGE, and p-NF-κB p65/NF-κB p65 elevated, while the 6-Keto-prostaglandin F1α (6-Keto-PGF1α) level decreased. Compared with the TAO group, the AP-L and AP-H groups showed reduced shedding of endothelial cells and mild thrombosis in the femoral artery tissue, with decreased whole blood viscosity, plasma viscosity, TXB2 level, and IL- 6, ICAM-1, VCAM-1, HMGB1, RAGE, and p-NF- κB p65/NF-κB p65 expression, and increased 6-Keto-

Objective: To investigate the effects of buddleoside on diabetic retinopathy and expression of nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) and NADPH oxidase 4 (NOX4) in diabetic rats. Methods: A diabetic model was established by intraperitoneal administration of streptozotocin. The buddleoside was administered at low and high doses (10 mg/kg and 40 mg/kg respectively) via intraperitoneal injection. Positive control group was injected intraperitoneally with recombinant human endostatin (5 mg/kg), while normal control group and model group were given normal saline (0.1 mL/100 g) by gavage. After continuous administration for 35 days, the histopathology of the rat’s retina was examined, and the levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-ɑ), vascular endothelial growth factor (VEGF), NLRP1, NOX4 and cysteinyl aspartate specific proteinase-1 (caspase-1) in the retinal tissues were detected. Results: Treatment with buddleoside and recombinant human endostatin improved the structure of the retina and increased the number of retinal ganglion cells, inner retinal thickness, and elevated the levels of GSH and SOD in retinal tissue. The levels of MDA, IL-1β, IL-6, TNF-ɑ, VEGF, NLRP1, NOX4 and caspase-1 in the retinal tissue were reduced. The effect in each buddleoside-treated group was dose-dependent, but less potent than that of the positive control group. Conclusion: Buddleoside can effectively protect the retinal ganglion cells of diabetic rats and improve the structure of the retina, which may be attributed to the inhibition of NLRP1 and NOX4 protein expression.

Objective: To investigate the effects of long non-coding RNA (lncRNA) NORAD on high glucose (HG)- induced proliferation, invasion, and epithelial-mesenchymal transition of trophoblast cells by targeting microRNA (miRNA)-199a/forkhead box O1 (FOXO1) axis. Methods: HTR-8/SVneo cells were induced with 30 mmol/L glucose and divided into five groups: HG group (non-transfected plasmid), si-NC group, si-NORAD group, si-NORAD+miR inhibitor-NC group, and si-NORAD+miR-199a inhibitor group. Normal HTR-8/SVneo cells cultured with 5.5 mmol/L glucose served as the normal glucose (NG) group. qRT-PCR was applied to measure the mRNA levels of NORAD, miR-199a, and FOXO1. The CCK-8, EdU staining, and colony formation assays were used to detect cell proliferation. Transwell assay was employed to detect cell migration and invasion. Western blotting was applied to analyze the protein expression of FOXO1, E-cadherin, Snail, and Vimentin. Dual-luciferase reporter assay and RNA immunoprecipitation were adopted to validate the targeting relationships among NORAD, miR-199a, and FOXO1. Results: Compared with the si-NC group, the expression of miR-199a, OD450 value, EdU-positive cell rate, numbers of migration and invasion cells, clone formation rate, the protein expression of Snail and Vimentin in the si-NORAD group increased, while the mRNA expression of NORAD and FOXO1, and the protein expression of FOXO1 and E-cadherin reduced. The miR-199a inhibitor reversed the effects of si-NORAD on the aforementioned indicators. There were targeting relationships among NORAD, miR-199a and FOXO1. Conclusion: Knockdown of NORAD may regulate the miR-199a/FOXO1 axis to promote proliferation, invasion, and epithelial-mesenchymal transition in HG-induced trophoblast cells.

Objective: To investigate the protective effects of salvianolic acid A (SAA) on imiquimod (IMQ)-induced psoriasis in mice by regulating the stimulator of interferon genes (STING)/nuclear factor kappa-B (NF-κB) signaling pathway. Methods: The mice were divided into five groups: model group, control group, low-dose SAA group, high-dose SAA group, and SAA+DMXAA (STING agonist) group. Serum levels of interleukin (IL)-23, IL-17A, IL-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) were measured by ELISA. The mRNA expression levels of IL-17A, CXC chemokine ligand (CXCL) 2, and CXCL1 in lesional skin tissues were detected by RT-qPCR. The protein expression levels of STING and NF-κB in lesional skin tissues were determined by Western blotting. Histopathological changes in lesional skin tissues were observed. Results: Compared with control group, the model group presented typical psoriasis-like skin lesions, with elevated, serum levels of IL-6, IL-23, TNF-α, IL-1β, IL-17A, mRNA expressions of IL-17A, CXCL2, and CXCL1 in tissues, as well as protein expressions of STING and p-NF-κB/NF-κB. Compared with the model group, the low and highdose SAA groups showed alleviated skin lesions, with downregulation of all quantitative indicators. Compared with the high-dose SAA group, the SAA+DMXAA group exhibited aggravated skin lesions, with upregulation of all quantitative indicators. Conclusion: SAA can alleviate IMQ-induced psoriasis lesions in mice by regulating the STING/NF-κB signaling pathway, thereby exerting a protective effect.

Objective: To estimate the sex based on hand dimensions among university students in the northern part of Henan Province by using discriminant analysis. Methods: A total of 955 college students (629 females and 326 males, aged 20–24 years) were examined. Hand length, breadth and hand index of each participant were measured. Statistical analyses were performed by using IBM SPSS 26.0. Results: All measured or derived hand parameters were significantly higher in males than in females. The hand dimensions showed higher accuracy than hand index in sex determination. Among all hand dimensions, the hand width had the highest accuracy in sex determination. The sex differences in the hand index on both sides were statistically significant. The hand variable showed pronounced gender dimorphism, and the hand index and palm index remained poor sex indicators. Conclusion: Hand length, hand width, and hand index all exhibit sex dimorphism and can be used for the determination of sex using hand alone. Hand width yielded the highest accuracy in sex determination, f ollowed by hand length and hand index.

In recent years, numerous studies have shown that exercise training can improve memory and cognitive function in Alzheimer’s disease (AD) patients, slow disease progression, and enhance their ability to perform daily activities independently, thereby providing a new perspective for the comprehensive treatment of AD. To more comprehensively address this complex neurodegenerative disease, this review systematically elucidates the mechanisms by which exercise training alleviates AD, specifically: exercise-induced epigenetic modification in AD; exercise inhibition of inflammatory responses in AD; exercise reduction of β-amyloid (Aβ) deposition and Tau protein hyperphosphorylation; exercise promotion of adult hippocampal neurogenesis and the secretion/release of growth factors; exercise regulation of gut microbiota diversity; exercise amelioration of mitochondrial dysfunction; and the relationship between exercise and cerebral blood flow.