Objective: To explore the effect of long noncoding RNA (lncRNA) small nucleolar RNA host gene 11 (SNHG11) on the proliferation and stemness of pancreatic cancer cells and its possible mechanisms. Methods: The expression of lncRNA SNHG11 in pan cancer and corresponding paracancerous tissues, pancreatic cancer and pancreatic tissues were analyzed by TCGA and GTEx databases; the expression of lncRNA SNHG11 in pancreatic cancer PANC1, PaTu8988 and MIApaca-2 cells was detected by qRT-PCR; pcDNA3.1 and pcDNA3.1-SNHG11 plasmids were transfected into PANC1 and PaTu8988 cells, respectively; sh-EGFP and sh-SNHG11 plasmids were transfected into PANC1 and MIApaca-2 cells, respectively. Plasmid transfection efficiency was measured by qRT-PCR, cell proliferation rate was measured by CCK8 assay, clone formation experiment was used to detect the number and size of cell clones, qRT-PCR and Western blotting were used to detect the mRNA and protein levels of stemness indicators, including NANOG homeobox protein (NANOG), sex determining region of Y-chromosome related HMG-box2 (SOX2), octamer binding transcription factor 4 (OCT4), cellular oncogene myc (c-myc) and LIN28 homolog A (LIN28A), respectively. Stem cell spheroidization experiment was used to detect changes in the number and diameter of stem cell spheroidization; RNA immunoprecipitation assay was used to detect the binding of SNHG11 and LIN28A. Co-ransfection of pcDNA3.1-SNHG11+sh-LIN28A or co-transfection of sh-SNHG11+3×FlagLIN28A, qRT-PCR was used to detect the mRNA levels of stemness indicators NANOG, SOX2, OCT4, and c-myc. Stem cell spheroidization experiments were used to detect changes in the number and diameter of stem cell spheroidization. Results: SNHG11 is highly expressed in a variety of malignant tumors, including pancreatic cancer. The relative expression of lncRNA SNHG11 in pancreatic cancer cells from low to high was PaTu8988, PANC1, MIApaca-2 (P<0.05). Up-regulating of SNHG11 expression significantly increased the proliferation and stemness of pancreatic cancer cells, while down-regulating exerted the opposite outcomes (P<0.05). LncRNA SNHG11 could bind to LIN28A. Overexpression of SNHG11 significantly increased the stemness of pancreatic cancer cells, and interference with LIN28A greatly reduced the stemness of pancreatic cancer cells, downregulating LIN28A in pancreatic cancer cells with over-expression of SNHG11 markedly inhibited the stemness promotion of pancreatic cancer cells (P<0.05). By silencing SNHG11, the stemness ability of pancreatic cancer cells was significantly reduced, while overexpression of LIN28A significantly enhanced the stemness ability of pancreatic cancer cells. Upregulating the expression of LIN28A in pancreatic cancer cells with low expression of SNHG11 could significantly reverse the stemness inhibition of pancreatic cancer cells (P<0.05). Conclusion: LncRNA SNHG11 may promote the proliferation and enhance the stemness of pancreatic cancer cells by up-regulating the expression of LIN28A.

Objective: To investigate the effect of O-linked N-acetylglucosamine (O-GlcNAc) glycosylation on the adhesion between gastric cancer cells and extracellular matrix and its potential mechanisms. Methods: The mRNA expressions of glutamine-fructose-6-phosphate transaminase 1 (GFPT1), glucosamine-phosphate N-acetyltransferase 1 (GNPNAT1), phosphoglucomutase 3 (PGM3), UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1), N-acetylglucosamine transferase (OGT) and O-GlcNAcase (OGA) in pan-cancer and human gastric cancer tissues were analyzed by UALCAN database. The relative mRNA expression levels of GFPT1, GNPNAT1, PGM3, UAP1, OGT and OGA in gastric cancer HGC-27, AGS, and SNU-1 cells were analyzed by DepMap database. The relative expression levels of GFPT1, OGT and OGA proteins and the glycosylation modification levels of O-GlcNAc in human gastric mucosal epithelial GES-1 cells and gastric cancer HGC-27, AGS and SNU-1 cells were detected by Western blotting. The signaling pathways enriched by the gastric cancer samples with differential expression of OGT and OGA were explored by Gene Set Enrichment Analysis (GSEA). The adhesion ability of human gastric mucosal epithelial GES-1 cells and gastric cancer HGC-27, AGS, and SNU-1 cells was detected by cell adhesion assay. OGT inhibitor OSMI-1 and OGA inhibitor Thiamet G (TMG) were used to decrease or increase O-GlcNAc glycosylation modification levels of gastric cancer cells HGC-27 and SNU-1 cells, respectively, and their adhesion ability was detected by adhesion assay. Results: The mRNA expression levels of GFPT1, GNPNAT1, PGM3, OGT, and OGA were significantly elevated in pan-cancer tissues, especially gastric cancer, compared with those in adjacent tissues (P<0.05). Compared with HGC-27 and SNU-1 cells, the mRNA expressions of GFPT1, GNPNAT1, PGM3 and UAP1 in AGS cells of gastric cancer were higher, but the mRNA expressions of OGT and OGA were lower. The expression of GFPT1 protein was not significantly different among human gastric mucosal epithelial cells and three gastric cancer cell lines, but the expression of OGT and OGA protein was significantly increased in HGC-27 and SNU-1 cells compared with human gastric mucosal epithelial GES-1 cells (P<0.01). The relative expression level of O-GlcNAc in SNU-1 cells was significantly higher than that in GES-1 cells and HGC-27 and AGS cells (P<0.01). GSEA enrichment analysis indicated that OGT and OGA may regulate the adhesion ability of gastric cancer cells by regulating the level of O-GlcNAc modification. Decreasing the O-GlcNAc modification level of gastric cancer cells could significantly reduce the adhesion ability of gastric cancer cells. Conclusion: The level of O-GlcNAc glycosylation in gastric cancer HGC-27 and SNU-1 cells was significantly increased, which may enhance the adhesion ability of gastric cancer HGC-27 and SNU-1 cells by affecting the expression of related adhesion proteins in HBP.

Objective: To investigate the expression levels and clinical significance of long non-coding RNA (lncRNA) dynein axonemal heavy chain 17-antisense strand 1 (DNAH17-AS1) in gastric cancer tissues, as well as its effect on the biological function of gastric cancer cells and its possible mechanisms. Methods: Real-time quantitative fluorescent PCR (qRT-PCR) was used to detect the expression levels of lncRNA DNAH17-AS1 in tissue samples (108 gastric cancer tissues and corresponding adjacent tissues) and plasma samples (25 gastric cancer patients and 25 healthy people, respectively). Statistical analysis was carried out to analyze the correlation between the expression of DNAH17-AS1 and clinicopathological features of patients with gastric cancer. Gastric cancer AGS and HGC-27 cells were divided into siNC control group and siRNA knockout group, respectively, and transfected with si-NC and si-DNAH17-AS1 by liposome transfection method. The proliferation, cloning, migration and invasion of AGS and HGC-27 cells were detected by cell counting, plate cloning assay and Transwell assay, respectively. mRNA and protein expression levels of related genes in gastric cancer cells were detected by qRT-PCR and Western blotting, respectively. Results: Compared with adjacent tissues, the relative expression level of lncRNA DNAH17AS1 in gastric cancer tissues was significantly increased (P<0.001). The relative expression of lncRNA DNAH17AS1 in gastric cancer tissues was correlated with TNM stage and lymph node metastasis (P<0.05). The 5year survival rate of patients with high DNAH17-AS1 expression was significantly lower than that of patients with low DNAH17-AS1 expression (P<0.05). The relative expression level of lncRNA DNAH17-AS1 in plasma of patients with gastric cancer was significantly higher than that of healthy subjects (P<0.01); and the area under the ROC curve of plasma DNAH17-AS1 was 0.721 (P<0.01). Compared with the si-NC control group, the proliferation, cloning, migration and invasion of gastric cancer AGS and HGC-27 cells in the siRNA knockout group were significantly decreased (P<0.001); and the relative expression of Ecad mRNA and protein was significantly increased (P<0.01), while the mRNA and protein expression of N-cad, transcription factors Slug, Snail, ZEB1, MMP9, and β-catenin was greatly decreased (P<0.05 or P<0.01). Conclusion: LncRNA DNAH17AS1 is highly expressed in cancer tissues and plasma of patients with gastric cancer, which is associated with poor prognosis of patients. The decrease of DNAH17-AS1 expression could inhibit the proliferation, clone formation, invasion and migration of gastric cancer cells, which may be related to the regulation of epithelial mesenchymal transformation related molecules expression.

Objective: To investigate the effects of Diosgenin on the proliferation and invasion of gastric cancer cells and its underlying mechanisms. Methods: Gastric cancer MKN-28 and BGC-823 cells were selected and divided into the following groups: control group (routine culture), solvent group (medium supplemented with ethanol at the same volume as the Diosgenin group), and Diosgenin group (medium supplemented with final concentrations of 0.5, 2, and 8 μg/mL Diosgenin). The relative expression of inhibitory gastric cancer EMT-related peptide (IGCE) was detected by quantitative real-time PCR (qRT-PCR). Additionally, gastric cancer cells were divided into control group (routine culture), Lv-control group (medium supplemented with recombinant lentivirus Lv-control), and Lv-IGCE group (medium supplemented with recombinant lentivirus Lv-IGCE). At 72 hours postviral infection, cell proliferation activity was assessed using CCK-8, and cell invasion was evaluated with Transwell assay. For another experiment, gastric cancer cells were divided into control group (routine culture) and transfection group (cells transfected with pcDNA-IGCE-aa-GFP). At 48 hours post-transfection, green fluorescent protein (GFP) expression was observed under a microscope. MKN-28 cells transfected with pcDNA-IGCE-aa-GFP were further divided into control group (routine culture), solvent group (medium supplemented with ethanol at the same volume as the Diosgenin group), and Diosgenin group (medium supplemented with final concentrations of 0.5, 2, and 8 μg/mL Diosgenin). After 48 hours of coincubation with Diosgenin, IGCEaaGFP fusion protein expression was detected by Western blotting. Finally, gastric cancer cells were divided into control group (routine culture) and Lv-IGCE-smORF group (medium supplemented with lentivirus Lv-IGCE-smORF). At 48 hours postinfection, cells were treated with cycloheximide (CHX), and Krüppel-like factor 4 (KLF4) protein expression was detected by Western blotting at 0, 0.5, 1, 2, 4, and 8 hours posttreatment. Results: qRT-PCR results showed that, compared with the control group, the relative expression of IGCE in MKN-28 and BGC-823 cells treated with Diosgenin was significantly increased (P<0.05 or P<0.01), in a concentration-dependent manner. Functional assays revealed that, compared with the control group, the proliferation and invasion of MKN-28 and BGC-823 cells in the Lv-IGCE group were markedly inhibited (all P<0.01). The microscopic observation results showed that compared with the control group, there were obvious GFP fluorescence signals in MKN-28 and BGC-823 cells in the transfection group. Western blotting results indicated that IGCE-aa-GFP expression in Diosgenin-treated MKN-28 cells was significantly enhanced (all P<0.01), also in a concentration-dependent manner. After CHX treatment, KLF4 protein expression in Lv-IGCE-smORF group MKN-28 cells was higher than that in the control group at 0.5, 1, 2, 4, and 8 hours (P<0.05 or P<0.01). Conclusion: Diosgenin may inhibit the proliferation and invasion of gastric cancer cells by up-regulating IGCE-aa expression and inhibiting the degradation of KLF4 protein.

Objective: To investigate the expression of microRNA-1184 (miR-1184) in MGC-803 cells infected by Helicobacter pylori (Hp), and its effects on the proliferation, metastasis and inflammation of MGC-803 cells infected by Hp. Methods: Fluorescence quantitative PCR was used to detect the expression of miR-1184 in Hp-infected MGC-803 cells. MGC-803 cells were divided into control group (miR-NC group), miR-1184 mimics group (miR-1184 group), Hp infection group (miR-NC+Hp group), miR-1184 mimics combined with Hp infection group (miR-1184+Hp group), inhibitor control group (In-NC group) and miR-1184 inhibitor group (anti-miR-1184 group). The effects of miR-1184 and Hp infection on the proliferation, cell cycle, metastasis and expression levels of related proteins of gastric cancer cells were detected by CCK-8 assay, colony formation assay, cell cycle assay, Transwell assay, wound healing assay and Western blotting assay. Results: Compared to the uninfected group, Hp infected MGC-803 cells produced cytopathic effect, and decreased the expression of miR-1184 (P<0.01). After transfection of miR-1184 mimics in MGC-803 cells, miR-1184 expression increased, cell activity decreased, colony formation ability decreased, G1 phase cells increased, S phase and G2 phase decreased, and wound healing ability decreased. In addition, miR1184 mimics up-regulated the expression level of E-cadherin, decreased the expression level of N-cadherin and Vimentin, inhibited the process of epithelial-mesenchymal transformation (EMT) (P<0.05 or P<0.01). Compared with miR-NC+Hp group, miR-1184+Hp group inhibited MGC-803 cell proliferation, cell cycle, migration ability and EMT effects (P<0.05 or P<0.01). Compared with miR-NC+Hp group, miR-1184+Hp group decreased the mRNA expression of TNF-α, IL-1β, IL-6 and IL-8 in MGC-803 cells, and inhibited the proportion of NF-κB p-p65/NF-κB p65 protein expression (P<0.05). Furthermore, miR-1184 inhibitors promoted the proliferation, cell cycle, cell transfer and EMT process of MGC-803 cells (P<0.05 or P<0.01). Conclusion: The expression of miR-1184 is decreased in Hp-infected gastric cancer cells, and the high expression of miR-1184 reversed the effects of Hp infection on the proliferation, migration and inflammatory response of gastric cancer cells.

Objective: To investigate the regulatory roles of farnesoid X receptor (FXR) and takeda G-protein coupled receptor 5 (TGR5) in cancer-associated fibroblasts (CAFs) on T-cell infiltration in hepatocellular carcinoma and their impact on patient prognosis. Methods: Multiplex fluorescence immunohistochemistry (mIHC) was performed on hepatocellular carcinoma tissue microarrays and paired adjacent non-tumor tissues. The HALO digital pathology image analysis system was used to quantitatively assess the expression levels of CD4, CD8, alpha-smooth muscle actin, TGR5, and FXR, obtaining fluorescence intensity values. Bioinformatics and statistical analyses were performed using R software. Results: Analysis using mIHC combined with the HALO digital pathology system revealed no significant correlation between number of CAFs and either T cell infiltration or overall survival in HCC patients (P>0.05). However, low FXR expression in CAFs was associated with significantly reduced T cell infiltration and poorer overall survival compared to high FXR expression (P<0.01). Conversely, high TGR5 expression in CAFs correlated with markedly lower T cell infiltration and worse overall survival than low TGR5 expression (P<0.01). Conclusion: FXR and TGR5 in tumor-associated fibroblasts may play crucial roles in regulating T cell infiltration and influencing the prognosis of hepatocellular carcinoma.

Objective: To study the effects of circ_0022382 on the proliferation and migration of breast cancer cells and the possible underlying mechanism. Methods: ① Screening for circRNAs that are highly expressed in breast cancer. Firstly, the differential analysis of the circRNArelated GEO dataset (GSE165884) and miRNA-related GEO dataset (GSE45498) of breast cancer were carried out, respectively, and then the miRNAs bound by the differentially expressed circRNAs were predicted by the ENCORI database, and finally the intersecting miRNAs and the miRNAs predicted by the GEO database were obtained by the Venn diagram. ② The complementary DNA (cDNA) and genomic DNA (gDNA) derived from MDA-MB-231 of breast cancer cells were PCR with convergent primers and divergent primers, respectively, and the PCR products were separated by agarose gel electrophoresis to verify the circular structure of the circ_0022382. ③ Realtime quantitative PCR (qRT-PCR) was used to detect circ_0022382 expression in breast cancer cells MDA-MB-231 and MCF-7 and normal breast epithelial cell MCF-10A, as well as in breast cancer tissues and adjacent tissues, respectively. ④ MDA-MB231 cells and MCF-7 cells were transfected with siNC and sicirc_0022382, qRT-PCR was used to detect transfection efficiency, clone formation assay and scratch healing assay were used to detect the proliferation and migration of breast cancer cells. qRT-PCR was used to detect the expression level of let-7a-5p. The si-circ_0022382 group was transfected with let-7a-5p inhibitor NC and let-7a-5p inhibitor, respectively, and the proliferation and migration of breast cancer cells were detected by clone formation assay and scratch healing assay. ⑤ The downstream signaling pathway of let-7a-5p was analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. MDA-MB-231 cells and MCF-7 cells were transfected with let-7a-5p NC and let-7a-5p mimic, respectively, and the expression of PI3K/AKT/mTOR signaling pathway-related proteins was detected by Western blotting and cellular immunofluorescence, and the expression of p-AKT in breast cancer cells co-transfected with let-7a-5p inhibitor and sicirc_0022382 was detected by Western blotting. Results: ① GSE165884 differential analysis yielded 37 high-expressing circRNAs, GSE45498 differential analysis yielded 6 low-expressing miRNAs, and among the 37 high-expression circRNAs, circ_0022382 corresponding miRNAs and 6 lowexpressing miRNAs had intersecting gene let-7. ② cDNA derived from breast cancer cell MDA-MB-231 can be amplified by both divergent primers and convergent primers, whereas gDNA can only be amplified by convergent primers. ③ Compared with MCF-10A, circ_0022382 was significantly more expressed in MDA-MB-231 and MCF-7 (P<0.001). The expression of circ_0022382 in breast cancer tissues was also higher than that in adjacent tissues (P<0.05). ④ After knocking down circ_0022382, the proliferation and migration of MDA-MB-231 and MCF-7 cells decreased significantly (P<0.001), and the expression level of let-7a-5p increased significantly (P<0.01 and P<0.05). Co-transfection of let-7a-5p inhibitor significantly rescued the decreased proliferation and migration of MDA-MB-231 and MCF-7 cells transfected with si-circ_0022382 (P<0.01 and P<0.001). ⑤ The results of KEGG signaling pathway enrichment analysis showed that PI3KAKT and mTOR signaling pathways were significantly enriched. Compared with the let-7a-5p NC group, the expression of p-AKT, p-PI3K and mTOR proteins in the let-7a-5p mimic group significantly decreased. Co-transfection of let-7a-5p inhibitor significantly rescued the decrease of p-AKT expression of MDA-MB-231 and MCF-7 cells transfected with si-circ_0022382. Conclusion: Circ_0022382 is highly expressed in breast cancer and promotes the proliferation and migration of breast cancer cells by targeting the let-7a-5p/PI3K/AKT/mTOR axis, which may be a potential target for breast cancer treatment and diagnosis.

Objective: To clarify the effect of YAN′s Wenyang Huayu Lishui Formula (YWHLF) on improving cardiac function in rats with chronic heart failure (CHF) and to explore its mechanisms. Methods: The CHF rat model was established through abdominal aortic constriction surgery (AAC) and treated with low (182 mg/g) and high (363 mg/g) doses of YWHLF for 8 weeks. Hemodynamic indicators were measured using Doppler echocardiography, myocardial histopathology was analyzed with HE staining, myocardial cell apoptosis was detected using TUNEL staining, and plasma levels of Btype natriuretic peptide (BNP), reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and cardiac troponin T (cTnT) were measured by ELISA. Additionally, the protein expression of nuclear factor E2related factor 2 (NRF2), heme oxygenase1 (HO1), and NAD(P)H quinone oxidoreductase 1 (NQO1) in myocardial tissue was detected by Western blotting. Pretreat H9c2 cells with 3% and 8% (v/v) YWHLF and 5 μmol/L NRF2 inhibitor ML385 for 1 h, followed by 24 h of oxygenglucose deprivation (OGD) treatment. Cell proliferation activity was measured using the CCK8 assay, and expression of NRF2 protein in nuclear was detected by Western blotting. Additionally, the effect of YWHLF on the binding efficiency of NRF2 protein to antioxidant response elements (ARE) in H9c2 cells was analyzed using the luciferase assay. Results: YWHLF administration increased left ventricular systolic pressure (LVSP) and the maximum rate of left ventricular pressure rise (or fall) (±dp/dtmax) in CHF rats, reduced left ventricular enddiastolic pressure (LVEDP), decreased inflammatory infiltration and cell apoptosis in myocardial tissue, reduced plasma levels of BNP, ROS, MDA, and cTnT, and increased SOD levels. Furthermore, YWHLF upregulated the expression of NRF2, HO1, and NQO1 proteins in myocardial tissue, with the highdose group showing significantly better effects than the lowdose group. Cellular experiments indicated that YWHLF pretreatment promoted the proliferation of H9c2 cells under OGD conditions and increased nuclear translocation of NRF2 protein, with the highdose group showing a significantly stronger effect than the lowdose group. The ML385 reversed the effects of YWHLF. YWHLF significantly enhanced the binding of NRF2 protein to ARE in H9c2 cells. Conclusion: YWHLF can improve cardiac function in CHF rats by activating the NRF2 signaling pathway and inhibiting ischemiainduced myocardial injury.

Objective: To improve the isolation and culture methods of primary cardiomyocytes in neonatal mouse to stably get high activity and purity of primary mouse cardiomyocytes. Methods: Hearts from 1-3day-old neonatal mouse were harvested, and a two-step digestion method combining trypsin and type Ⅱ collagenase was used. Differential adherence was then employed to purify the cardiomyocytes, which were cultured in vitro in modified Dulbecco′s Modified Eagle Medium (DMEM) containing 10% horse serum. The morphological characteristics and beating patterns of the cardiomyocytes were observed under an inverted phase-contrast microscope. Cell viability was assessed by using trypan blue staining, and cardiomyocytes were identified by immunofluorescence staining for cardiac troponin T (cTnT). Results: After 24 hours of culture, the primary neonatal mouse cardiomyocytes adhered to the culture dish and exhibited spontaneous beating. By 96 hours, the cells aggregated into clusters and displayed synchronous beating. The cell survival rate was 90.63%, and the purity of cardiomyocytes reached 93.17%. Conclusion: The modified isolation and culture methods used in this study obtained the primary neonatal mouse cardiomyocytes with high activity and purity.

Objective: To investigate the association between plasma lowdensity lipoprotein (LDL) subtypes and posterior circulation ischemia (PCI) in the middleaged and elderly patients. Methods: A total of 168 middle-aged and elderly PCI patients who were treated in the Affiliated People′s Hospital of Jiangsu University from January 2023 to May 2024 were included. According to the infarct changes of magnetic resonance diffusion-weighted imaging, they were divided into transient ischemic attack (TIA) group (83 cases) and cerebral infarction group (85 cases). In addition, 81 middle-aged and elderly healthy subjects who underwent physical examination in the hospital during the same period were randomly selected as the control group. The general data of the three groups were collected, and the levels of high-density lipoprotein, intermediate density lipoprotein, and LDL subtypes were measured. Logistic regression analysis was used to analyze the influencing factors of posterior circulation ischemia in middle-aged and elderly patients. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of plasma LDL-3 and LDL-4 levels for PCI in middle-aged and elderly patients. Results: Compared with the control group, the levels of plasma LDL-3 and LDL-4 and the proportion of hypertension in TIA group and cerebral infarction group were significantly increased (P<0.05), while the level of LD-1 was significantly decreased (P<0.05). Compared with the control group, the proportion of smoking and diabetes, and the level of triacylglycerol (TG) were significantly increased in cerebral infarction group (P<0.05). Logistic regression showed that hypertension（OR=2.114, 95%CI: 1.024-4.364, P<0.05）, plasma LDL-3（OR=1.006, 95%CI: 1.003-1.009, P<0.001) and LDL-4 （OR=1.007, 95%CI: 1.002-1.013, P<0.05） were independent risk factors for PCI in middle-aged and elderly patients, while plasma LDL-1 （OR=0.402, 95%CI: 0.201-0.806, P<0.05） was an independent protective factor for PCI. The area under the ROC curve (AUC) of plasma LDL-3 and LDL-4 was 0.847 and 0.798, the sensitivity was 86.9% and 70.8%, and the specificity was 72.8% and 86.4%, respectively. The AUC of the combined diagnosis of the two indicators was 0.853. Conclusion: The increase of plasma LDL-3 and LDL-4 levels and the decrease of LDL-1 levels are related to the occurrence of PCI.

Objective: To investigate key microRNAs (miRNAs) and mRNAs involved in the pathogenesis of Parkinson′s disease (PD) through bioinformatics analysis and clinical experiments, and identify mRNAs and miRNAs with diagnostic and therapeutic potential. Methods: mRNA and miRNA sequencing data from the Parkinson′s progression markers initiative (PPMI) were analyzed to identify differentially expressed genes (DEGs). Machine learning algorithms were applied to determine critical miRNAs. Target genes of these miRNAs were predicted using multiple miRNA-mRNA interaction databases. Predicted targets were crossmatched with DEGs to construct an miRNA-mRNA regulatory network. Protein-protein interaction (PPI) networks were built by using the STRING database and Cytoscape software to identify hub genes. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed on clinically collected samples to validate the identified hub genes and miRNAs. Results: Through differential gene expression analysis (DEGA) combined with machine learning, two key miRNAs-miR-214 and miR-421-were identified. Further analysis using weighted gene co-expression network analysis (WGCNA), miRNA-mRNA interaction databases, and PPI network screening revealed STAT3 as the hub gene. Clinical validation confirmed the correlation between miR-214 and STAT3 expression, demonstrating their diagnostic efficacy. Conclusion: MiR-214 may suppress STAT3 expression, thereby alleviating aberrant aggregation of α-synuclein, inflammatory responses, and oxidative stress in PD pathogenesis. Both miR-214/STAT3 demonstrated strong diagnostic predictive efficacy in clinical samples and exhibited potential as therapeutic targets for PD.