Objective To characterize the expression profiles and potential biological functions of circular RNAs (circRNAs) in peripheral blood mononuclear cells (PBMCs) from patients with acute ischemic stroke (AIS), and to evaluate their clinical relevance for diagnostic and prognostic applications. Methods Transcriptome-wide RNA sequencing was performed on PBMC samples collected from AIS patients (n=5) and age- and sex-matched healthy controls (n=5) in a discovery cohort to delineate the circRNA landscape in AIS. The top ten differentially expressed circRNAs (DECs) were identified based on fold change and statistical significance. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted to explore the biological functions of the host genes of these DECs. Validation of candidate circRNAs was performed using Real-time PCR in an independent validation cohort consisting of 30 AIS patients and matched controls. The diagnostic and prognostic significance of validated circRNAs was further assessed through correlation analyses with National Institutes of Health Stroke Scale (NIHSS) scores and receiver operating characteristic (ROC) curve analysis. Results  Functional enrichment analysis revealed that the host genes of the top DECs were predominantly involved in immune responses, protein degradation and cell death. Among the candidates, hsa_circ_0075436 and hsa_circ_0005729 were significantly downregulated in AIS patients as validated by Real-time PCR. Importantly, their expression levels were inversely correlated with NIHSS scores at both admission and discharge, suggesting a potential link to disease severity and neurological recovery. ROC analysis demonstrated that both circRNAs exhibited robust diagnostic performance, highlighting their potential as blood-based biomarkers. Conclusion The host genes of DECs are involved in key pathways related to transcriptional regulation, protein ubiquitination, immune response, protein degradation, and apoptosis. Among them, hsa_circ_0075436 and hsa_circ_0005729 represent promising candidates for further development as diagnostic and prognostic biomarkers in acute ischemic stroke.

Objective  To investigate the relationship between the expression changes of synaptosomal-associated protein 25 (SNAP25) and the onset of amyotrophic lateral sclerosis (ALS), explore the neuroprotective effect of SNAP25 on ALS motor neurons, and provide potential therapeutic targets for ALS.  Methods  Differentially expressed genes in ALS transgenic mice were screened using public databases. Focusing on SNAP25, 36 pairs of hSOD1-G93A transgenic ALS mice and their wild-type (WT) littermates at different disease onset time points were selected as animal models. hSOD1-G93A mutant and hSOD1-WT NSC34 cells were generated as ALS cell models. Real-time PCR and Western blotting were used to verify differential expression in vivo and in vitro. Immunofluorescence assays were performed to detect the co-localization of SNAP25 with neuronal marker neuronal nuclei (NeuN) in mouse spinal cords. SNAP25 was overexpressed in hSOD1-G93A mutant NSC34 cells, and cell viability was measured using a cell viability assay kit. After neurite induction, changes in neurite growth were analyzed, along with the expression of neurite-related proteins βⅢ-tubulin and growth associated protein 43（GAP-43). Results  Analysis of public databases revealed that SNAP25 expression was significantly lower in the spinal cord of ALS transgenic mice compared to WT mice. Animal studies confirmed that, compared to WT mice, the levels of SNAP25 protein and mRNA in the spinal cord of ALS mice decreased; SNAP25 co-localized with NeuN in the mouse spinal cord, and the integrated optical density of SNAP25 in SNAP25-positive neurons in ALS mice decreased. In vitro experiments revealed that the expression levels of SNAP25 protein and mRNA in hSOD1-G93A mutant NSC34 cells were reduced, and overexpression of Snap25 improved cell viability and promoted neurite outgrowth. Conclusion  The down-regulation of SNAP25 expression is closely related to the pathological process of ALS. Overexpression of Snap25 enhances the viability of hSOD1-G93A mutant NSC34 cells, promotes neurite outgrowth, thereby exerting a neuroprotective effect.

Objective  To investigate the distribution patterns of pain- and itch-specific neurons within the lateral parabrachial nucleus (LPB), and to map their divergent downstream projection pathways. Methods  The male C57BL/6 mice were used, the optimal induction window for the tetracycline-off gene expression system (Tet-off system) was determined to be seven days through preliminary experiment (n=24). We then employed the Tet-off system, in combination with c-Fos promoter-driven viral vectors and Fos immunofluorescence histochemistry, to specifically label LPB neurons activated by pain (LPB^pain) or itch (LPB^itch) and to analyze their co-localization within LPB subnuclei (n=7). Finally, using Tet-off-based viral anterograde tracing (n=6), we compared the downstream projection targets of these two neuronal populations. Results  Sequential application of distinct noxious stimuli within defined time windows induced robust co-expression of viral vector-derived enhanced green fluorescent protein (EGFP) and Fos protein (labeled by red fluorescence) in the LPB. Microscopic analysis revealed that LPB^painand LPB^itch neurons were extensively co-localized in the external part of the lateral parabrachial nucleus (LPBE). The co-localization rate reached up to 72.9% for EGFP^＋ pain neurons with Fos^＋ itch neurons, and 79.5% for EGFP^＋ itch neurons with Fos^＋ pain neurons. Co-labelling rates in other subnuclei were relatively lower. Anterograde tracing results showed that both LPB^pain and LPB^itch neurons projected predominantly to overlapping regions, including the central amygdaloid nucleus (CeA), periaqueductal gray (PAG), zona incerta (ZI), revealing a highly similar overall projection pattern. Conclusion  While the LPB is activated by both pain and itch stimuli, its subnuclei exhibit distinct response profiles to these stimuli. The largely overlapping downstream projections of LPB^pain and LPB^itch neurons suggest that differential central encoding of pain and itch may rely more on molecular or functional mechanisms than on strictly segregated anatomical pathways.

Objective  To explore the morphological feature of the medial prefrontal cortex (mPFC) to nucleus reuniens (Re) projection pathway in mice with neuropathic pain. Methods  Twelve C57BL/6 mice were divided into 3 groups according to the method employed. Retrograde tracer fluoro-gold (FG) was injected into the Re of mice subjected to spared nerve injury (SNI, n=3) or sham (n=3) operation. The distribution and pain responsivity of retrogradely-labeled neurons in the mPFC were observed with c-Fos immunofluorescent staining. Red retrobeads (Beads) were injected into the Re of normal mice (n=3), and the neurochemical feature of the mPFC-Re projection neurons was examined with calmodulin-dependent protein kinase Ⅱ (CaMKⅡ) immunofluorescent staining. The c-Fos promoter-dependent anterograde virus was injected into the mPFC of SNI mice (n=3). The distribution of pain responsive mPFC-Re projection terminals and the target cell type were observed by calretinin (CR) immunofluorescent staining. Results  The FG or Beads retrogradely-labeled neurons were mainly distributed in the deep layers (Ⅴ and Ⅵ) of cingulate cortex area 1 (Cg1), prelimbic (PrL), and infralimbic (IL) cortices of the mPFC. About 96.81% of labeled neurons showed CaMKⅡ immunoreactivities. SNI caused a significant decrease in the c-Fos expression and the number of FG/c-Fos double-labeled neurons, mostly strikingly in the PrL. The c-Fos-driven anterograde virus labeled terminals were mainly distributed in the ventrolateral region of the Re and formed close contacts with CR-positive neurons. Conclusion  The projection neurons in the mPFC-Re pathway are dominantly excitatory neurons, and target on CR-positive neurons. Neuropathic pain results in the reduced activity of the mPFC-Re projection pathway.

Objective  To explore the ameliorative effect of chelated zinc on neuronal pyroptosis during the acute phase of cerebral ischemia in mice. Methods  Thirty adult male Kunming mice (25-30 g) were randomly divided into three groups according to the randomized controlled principle, sham operation group (sham), middle cerebral artery occlusion (MCAO) reperfusion 3 days group (MCAO), and zinc chelation Tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) group, with 10 mice in each group. Mice in each group were anesthetized by inhalation, and MCAO model was established by the suture-occlusion method. Mice in each group were subjected to 1 hour of ischemia followed by 3 days of reperfusion. Mice in the TPEN group were intraperitoneally injected with the zinc ion chelator TPEN at a dose of 10 mg/kg immediately after reperfusion, while mice in the sham group only underwent blunt separation of blood vessels. Neurological function scoring, 2,3,5-triphenyltetrazolium chloride (TTC) staining, and laser speckle imaging were used to evaluate the protective effect of zinc chelation on cerebral injury in mice. Transmission electron microscopy, immunofluorescent staining, zinc probe, and Western blotting were performed to observe the improvement effect of zinc chelation on neuronal pyroptosis after cerebral injury in mice. Transcriptome sequencing combined with Gene Ontology (GO) enrichment analysis, immunofluorescence co-staining, and Western blotting were used to detect the expression levels of absent in melanoma 2 (AIM2), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, cleaved-Caspase-1, and gasdermin (GSDM) family proteins GSDMD-N proteins after zinc chelation intervention. Results  Zinc chelation improved the neurological function scores of mice (P<0.05), reduced the cerebral infarct volume (P<0.01), and restored the cerebral blood perfusion (P<0.001). Immunofluorescence co-staining results showed that compared with the model group, zinc chelation reduced intracellular zinc ion accumulation in neurons, decreased the number of neuronal deaths in the ischemic hemisphere, and lowered the content of free zinc ions. Transmission electron microscopy results revealed a significant reduction in pyroptotic bodies in the cerebral cortex of mice in the zinc chelation group compared with the model group. Transcriptome sequencing with GO enrichment analysis indicated that inflammation and massive pyroptosis erupted on 3 days after cerebral ischemia. Western blotting and immunofluorescence results demonstrated that TPEN treatment reduced the expression of AIM2 inflammasome proteins induced by cerebral ischemic injury (P<0.05). Conclusion  Chelated zinc exerts a neuroprotection against neuronal pyroptosis in mice after acute cerebral ischemia.

Objective  To investigate the effect of wogonin (Wog) on liver fibrosis in rats with hepatitis B and the mechanism of regulating Hippo-Yes-associated protein (YAP) / transcriptional coactivator with PDZ-binding motif (TAZ) signaling pathway. Methods  Rats were randomly grouped into normal control (NC) group, model group（Mod）, low-dose Wog group (Wog-L 10 mg/kg), medium-dose Wog group (Wog-M 20 mg/kg), high-dose Wog group (Wog-H 40 mg/kg), and Wog-H+Hippo inhibitor group (40 mg/kg Wog+7 mg/kg XMU-MP-1), each with 12 rats. Except for the NC group, rat model was established using hepatitis B virus (HBV) in other groups, injected once a week for three consecutive weeks. ELISA assay was used to detect serum liver function indicators, aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamyl transpeptidase (GGT), hepatitis Be antigen (HBeAg), hepatitis B virus surface antigen (HBsAg), and inflammatory factors, transforming growth factor-β (TGF-β), platelet-derived growth factor (PDGF), and liver fibrosis indicators, type Ⅳ collagen (Col-Ⅳ), hyaluronic acid (HA), type Ⅲ procollagen peptide (PⅢP), and laminin (LN). Hydroxyproline (Hyp) detection kit for detecting Hyp levels. HE and Masson staining were used to detect the morphology and fibrosis of liver tissue. TUNEL method was used to detect cell apoptosis in liver tissue. Western blotting was used to estimate the expression of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Col-Ⅰ), and proteins related to Hippo-YAP/TAZ signaling pathway. Real-time PCR was used to detect the mRNA expressions of YAP and TAZ in liver tissues. Results  Compared with the NC group, the model group showed swelling, abnormal arrangement of cells, and obvious collagen fiber proliferation in the liver tissue, the levels of serum AST, GGT, ALT, HBeAg, HBsAg, TGF-β, PDGF, Col-Ⅳ, PⅢP, LN, HA, and the proportion of collagen fiber deposition in liver tissues, as well as the levels of Hyp, α-SMA, Col-Ⅰ protein expression, and the apoptosis rate obviously increased (P<0.05), and the protein and mRNA expression levels of YAP and TAZ increased significantly(P<0.05). Compared with the model group, the liver tissue damage of rats in each dose Wog group was reduced, the levels of serum-related indicators, and the proportion of collagen fiber deposition in liver tissues, as well as the levels of Hyp, α-SMA, Col-Ⅰ protein expression, and the apoptosis rate, and the protein and mRNA expression levels of YAP and TAZ significantly reduced (P<0.05). Compared with the Wog-H group, the Wog-H+XMU-MP-1 group showed obvious liver fibrosis damage (P<0.05). Conclusion  Wog may alleviate liver fibrosis in rats with hepatitis B by regulating activity of the Hippo-YAP/TAZ pathway.

Objective  To evaluate the alleviating effect of indole-3-carboxamide(WCL-3) on myocyte atrophy in vitro. Methods  An activin receptor ⅡB (ActRⅡB) kinase inhibition assay in vitro was established to assess the inhibitory activity of WCL-3. Based on the enzymatic inhibition result, WCL-3 anti-muscle atrophy and anti-adipocyte degradation effects were further investigated. Myotube diameter measurement and morphological observation were used to evaluate the protective effect of WCL-3 on myotube atrophy. Oil red O staining and quantitative analysis were conducted to assess the WCL-3 inhibitory effect on adipocyte degradation. Western blotting analysis was performed to detect the expression levels of p-Smad2/3, Smad2/3, p-Akt, Akt, Atrogin-1, and myogenic differentiation 1(MyoD1) in muscle cells. Results  Kinase inhibition assays demonstrated that WCL-3 exhibited potential inhibition of ActRⅡB kinase, half-maxinal inhibitory concentration (IC50)=4.1 nmol/L. In C2C12 myotube atrophy models induced by CT26 tumor-conditioned medium and transforming growth factor-β1（TGF-β1）, WCL-3 alleviated myotube atrophy in a dose-dependent manner. At the molecular level, WCL-3 downregulated the phosphorylation of Smad2/3 and inhibited the expression of Atrogin-1, while upregulating p-Akt and promoting MyoD1 synthesis, thereby attenuating muscle atrophy. Additionally, WCL-3 ameliorated CT26-conditioned medium-induced lipid degradation in mature 3T3-L1 adipocytes. Conclusion  WCL-3 significantly alleviates myocyte atrophy in vitro primarily by inhibiting the TGF-β/ActRⅡB signaling pathway. Therefore, WCL-3 represents a promising lead compound for the development of therapeutic agents targeting muscle wasting and fat loss.

Objective  To investigate the immunogenic cell death-related gene markers mediating immune infiltration and poor prognosis in lung adenocarcinoma. Methods  Based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases, we analysed the expression profile of immunogenic cell death (ICD) genes in lung adenocarcinoma (LUAD), screened for the set of differentially expressed ICD-related genes (ICD-RGs) by weighted gene co-expression network analysis (WGCNA), established the LASSO-Cox prognostic model and carried out the functional and drug sensitivity analysis; in vitro validation of the inhibitory effect of BI-2536 on the proliferation and migration of LUAD cells, and verification of the expression of ICD-RGs by Western blotting of three clinical samples. Results  Differential genes of ICD-related subtypes were mainly enriched in aminoacyl-transfer RNA(tRNA) biosynthesis, citric acid cycle, base excision repair, oxidative phosphorylation, protein export, spliceosome, antigen processing and presentation, B-cell receptor signaling pathway, chemokine signaling pathway, cytokine-cytokine receptor interactions, natural killer cell-mediated cytotoxicity, and T-cell receptor signaling pathway, |NES|>1, P<0.05; ICD-RGs were significantly associated with both prognosis and immune microenvironment in LUAD patients (all P<0.05); BI-2536 had an inhibitory effect on the proliferation, invasion, and migration of both A549 and H1650 cells, P<0.05; Nomograms developed through the network calculator had a better predictive mechanism, and both receiver operating characteristic(ROC) and decision curve analysis(DCA) over time showed that nomograms were better predictors of prognosis; ICD-RGs showed differential expression in LUAD tissues and normal tissues in Human Protein Atlas(HPA) and Western blotting. Conclusion  ICD-RGs are significantly associated with immunity and prognosis of LUAD, and BI-2536 can be used as a clinical drug to inhibit the growth migration and invasion of LUAD cells, mediate the ICD-RGs pathway and biofunctional activity. In addition, the newly developed nomogram provides a more convenient and accurate method to predict the overall survival of LUAD patients.

Objective  To investigate the role and mechanism of microRNA-let-7a-5p (miRAN-let-7a-5p) in regulating apoptosis and cell cycle arrest in gastric cancer cells via the mouse double minute 2 (MDM2) / tumor protein 53 (p53) signaling pathway. Methods  Twenty pairs of gastric cancer tissues and matched adjacent non-tumor tissues were collected to detect let-7a-5p expression. Bioinformatics databases were used to analyze the expression of let-7a-5p in gastric cancer and its correlation with patient prognosis. Potential target genes of let-7a-5p were predicted；Both overexpression and knockdown cell models of let-7a-5p were established. Cell proliferation, migration, apoptosis, and cell cycle distribution were assessed using the CCK-8 assay, wound healing assay, and flow cytometry, respectively. The targeting relationship between let-7a-5p and MDM2 was verified by a dual-luciferase reporter assay. Western blotting and Real-time PCR were performed to measure the expression of MDM2, p53, and downstream factors, including Cyclin D1,cellular myelocytomatosis oncogene product(c-myc）、tumor protein 53(p53）, Bcl-2 and Bax. Results  Real-time PCR results showed that let-7a-5p was significantly downregulated in gastric cancer tissues compared with adjacent normal tissues (P<0.05), and low let-7a-5p expression was associated with poor prognosis. Transfection with let-7a-5p mimics markedly increased let-7a-5p levels in human gastric cancer cells HGC-27(P<0.01), while let-7a-5p inhibitor reduced its expression in human gastric cancer cells AGS(P<0.05). Overexpression of let- 7a-5p inhibited proliferation and migration, promoting apoptosis and cell cycle arrest in HGC-27 cells. In contrast, knockdown of let-7a-5p enhanced proliferation and migration, suppressed apoptosis and cell cycle arrest in AGS cells. Target prediction and dual-luciferase reporter assays confirmed that let-7a-5p directly targeted MDM2 and negatively regulated its expression. Western blotting and Real-time PCR results demonstrated that let-7a-5p overexpression upregulated p53 and Bax, and downregulated MDM2, Bcl-2, CyclinD1, and c-myc. Conversely, let-7a-5p knockdown increased the expression of MDM2, Bcl-2, Cyclin D1, and c-myc, and decreased p53 and Bax levels. Conclusion  Let-7a-5p promotes apoptosis and cell cycle arrest in gastric cancer cells by suppressing the MDM2/p53 signaling pathway.

Objective  To investigate the molecular mechanism by which U2 snRNP-associated SURP domain containing protein (U2SURP) influences the proliferation of HCT116 colorectal cancer cells under high glucose conditions, and to examine the effect of metformin. Methods  Clinical samples from colorectal cancer (CC, n=8) and diabetes-CC(DCC, n=8)patients were collected. High-glucose cell model was established by culturing HCT116 cells with 33 mmol/L glucose for 8 days. Diabetic xenograft model was generated by inducing diabetes in C57BL/6 mice via streptozotocin (STZ) injection (120 mg/kg) and subsequent subcutaneous implantation of MC38 cells. U2SURP expression in tissues and cells were analyzed by Western blotting and Real-time PCR. The proliferation level of HCT116 cells was analyzed by CCK-8 and colony formation assays following U2SURP knockdown using short hairpin RNA(shRNA) and cell cycle progression was analyzed by flow cytometry. The upstream regulator of U2SURP was predicted by the SRAMP and RBP suite databases. The expression level of insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) was examined in diabetic mouse xenograft models and high glucose-cultured cell models. Overexpression and knockdown cell lines were constructed to validate their effects on the proliferation of colon cancer cells under high glucose conditions. Using the high-glucose cultured HCT116 cell model, treatment with metformin (0.0128 mol/L), detected the expression of U2SURP and its upstream factor, cell proliferation capacity, and analyzed the regulatory role of the Akt/glycogen synthase kinase-3 (GSK-3β) signaling pathway. Results  U2SURP expression was significantly upregulated in both diabetic colorectal cancer tissues and high glucose-cultured HCT116 cells. High glucose promoted colorectal cancer cell proliferation, whereas knockdown of U2SURP inhibited this pro-proliferative effect and induced S-phase cell cycle arrest. We next screened IGF2BP3 as an upstream regulator of U2SURP and confirmedits overexpression promoted the proliferation of colon cancer cells, while its knockdown reversed the pro-proliferative effect of high glucose. Furthermore, metformin inhibited the upregulation of U2SURP and IGF2BP3 under high glucose conditions, weakened cell proliferation capacity, and reduced Akt and GSK-3β phosphorylation. Similarly, Akt inhibition reversed the upregulation of U2SURP and IGF2BP3 by high glucose, while Akt activation attenuated the suppressive effect of metformin on these genes. Conclusion  Under high glucose conditions, IGF2BP3 promotes colon cancer cell proliferation by regulating U2SURP in an m6A-dependent manner. Metformin mitigates this pro-proliferative effect by inhibiting the Akt signaling pathway, thereby reducing the expression of U2SURP and IGF2BP3.

Objective  To investigate the effects of urantide on myocardial injury in ApoE^-/- mice by regulating the urotensin Ⅱ(UⅡ)/urotensin II receptor (UT) system and the Wnt/β-catenin(β-cat) signaling pathway. Methods ApoE^-/- atherosclerotic(AS) mice were randomly divided into the model group, simvastatin group, and urantide low dose group, urantide medium dose group, and urantide high-dose urantide groups (n=12 per group), while 12 C57BL/6 mice were used as the normal control group. Myocardial histopathological changes were observed using HE and Masson’s trichrome staining. Immunohistochemistry was performed to detect UⅡ and G protein couple receptor 14 (GPR14) protein expression in cardiac tissues, while immunofluorescent staining was used to assess β-cat expression. Western blotting was employed to measure the expression levels of Wnt-3a, β-cat, and components of the β-catenin degradation complex, glycogen synthase kinase-3β(GSK-3β), p-GSK-3β, activity protein C (APC), axis inhibition protein-2(Axin-2), and casein kinase-1α(CK-1α) in myocardial tissues. Results  The model group exhibited typical pathological changes, including myocardial edema, inflammatory cell infiltration, and collagen deposition. Protein levels of UⅡ, GPR14, Wnt-3a, β-cat, and p-GSK-3β increased significantly, while CK-1α, Axin-2, APC, and GSK-3β decreased markedly(P<0.05). Compared with the model group, the urantide treatment group showed significant alleviation of myocardial injury, along with reduced expression of UⅡ, GPR14, Wnt-3a, β-cat, and p-GSK-3β, and increased levels of CK-1α, Axin-2, APC, and GSK-3β (P<0.05). Conclusion  Urantide mitigates myocardial injury in ApoE^-/- atherosclerotic mice by modulating the UⅡ/UT system and the Wnt/β-cat signaling pathway.

Objective  To examine the impact of a 25-amino-acid N-terminal peptide (α-ANT) derived from aminoacyl-transfer RNA synthetase complex-interacting multifunctional protein 1 (AIMP1) on scar development and the process of wound healing following skin injury. Methods  The full-thickness incision wound or full-thickness excision wound models were constructed in SPF-grade CD-1 male mice (n=51) using a sterile scalpel or a 9 mm skin punch. After creating the full-thickness incision wound model, the mice were divided into the simple full-thickness incision wound model group (blank group, n=9), the full-thickness incision wound model + Pluronic gel group (n=9), and the full-thickness incision wound model + α-ANT treatment group (60 μmol/L, n=12). After creating the full-thickness excision wound model, the mice were divided into the full-thickness excision wound model + Pluronic gel group (control group, n=9) and the full-thickness excision wound model + α-ANT treatment group (60 μmol/L, n=12). Wound healing was assessed, and HE staining was used to observe pathological changes in granulation tissue. Western blotting and immunofluorescence were employed to measure the expression levels of inflammatory cytokines and growth factors. The tensile properties of the skin lesions after repair were evaluated using a multifunctional tensile tester. CCK-8 and Transwell chemotaxis assays were conducted to examine the effects of α-ANT on the proliferation and migration characteristics of primary neutrophils. Results   After topical application of exogenous α-ANT peptide, the peptide could accumulate at the wound edges within 2 hours following the establishment of full-thickness skin wound models in mice. Whether applied to linear (incisional) or circular (puncture) wounds, α-ANT peptide significantly accelerated wound healing, reduced scar tissue formation, and effectively promoted the restoration of dermal tissue structure and mechanical strength post-healing. Western blotting and immunofluorescence results demonstrated that α-ANT peptide enhanced ERK activation, inhibited P38 MAPK activation, reduced neutrophil infiltration, and stimulated growth factor synthesis. Conclusion  Regardless of the morphology of skin wounds, α-ANT peptides can significantly promote skin wound healing by modulating the dynamic balance of ERK-P38 MAPK to inhibit neutrophil infiltration and secretion during the inflammatory process.

Objective  To systematically compare the distribution characteristics of handedness in different populations worldwide through meta-analysis and Edinburgh Handedness Inventory (EHI) questionnaire, and to clarify the handedness distribution characteristics of northwestern Chinese populations (Ningxia and Gansu populations as samples) as well as their differences from other populations. Methods  Literatures were retrieved and screened from CNKI and PubMed for meta-analysis. Taking Ningxia and Gansu populations as the research objects, EHI questionnaire was used to collect handedness data. EpiData double entry was applied for data management, and χ² test and multiple logistic regression were used to analyze the distribution characteristics and influencing factors. Results  Meta-analyses had demonstrated that the prevalence of non-right-handedness was significantly lower in Asian populations compared to Western populations, and there were also significant geographical differences in handedness distribution. Univariate analyses of the Ningxia and Gansu cohorts revealed that the rate of left-handedness was significantly higher in males than in females, higher in individuals with middle school or vocational school education than in those with primary school or college education, and higher in individuals born in spring or summer (all P<0.05). Further multivariate logistic regression analysis had confirmed that being male and having middle school or vocational school education were independent risk factors for left-handedness. After pooling the present data,it was found that the overall rate of non-right-handedness in China was 8.8%, which was only higher than that in Japan and Sweden (P<0.001), but lower than that in 15 countries or regions such as Bulgaria (all P<0.05). The characteristics of handedness distribution in male and female groups were consistent with those of the general population. Conclusion  Handedness distribution shows certain regional and gender differences.The Ningxia and Gansu cohorts in China exhibit a distinctive handedness distribution pattern, and the left-handedness ratio in males is higher than that in females

Objective  To construct the experimental teaching system of ‘three combinations + four integrations’ and explore its practical effects in the experimental teaching of regional anatomy. Methods  The teaching model of ‘three combinations + four integrations’ was established. A total of 600 undergraduate studentsmajoring in clinical medicine from Chengdu Medical College (grades 2021-2022) were selected as the research subjects. The traditional experimental teaching method and the experimental teaching method of ‘three combinations + four integrations’ were applied respectively. The teaching effectiveness was evaluated through regular quizzes, end-of-course assessments, experimental evaluations, PBL-based assessments, and students’ subjective feedback on the course. Results  The scores of regular quizzes, final end-of-course examinations, digital specimen examinations and the subjective recognition of students in the experimental groups of the two grades were all better than those of students in the control groups (all P<0.05). Conclusion  The ‘three-combinations + four-integrations’ experimental teaching system has effectively enhanced students’ academic performance, autonomous learning ability, practical skills, clinical thinking, innovation ability, and comprehensive knowledge application. It is beneficial for improving the overall teaching effectiveness.