Objectives: To elucidate the potential mechanisms of electroacupuncture (EA) in restoring detrusor-bladder neck dyssynergia (DBND) following suprasacral spinal cord injury (SSCI). Methods: A total of 52 specific pathogen-free (SPF) grade famale Sprague-Dawley (SD) rats (10 – 12 weeks, 250 – 280 g) were randomly assigned to either a sham group (n = 12) or a spinal cord injury model group (n = 40). In the model group, DBND was induced through Hassan Shaker spinal cord transection at T10 level, with 24 rats meeting inclusion criteria and subsequently randomized into DBND group (n = 12) and EA intervention group (DBND + EA group, n = 12). After spinal shock recovery (day 19 after modeling), DBND + EA group received EA treatment at Ciliao (BL32), Zhongji (RN3), and Sanyinjiao (SP6) acupoints for 20 min per session at 10/50 Hz frequencies, once daily for 10 d. Sham and DBND groups received anesthesia only without EA intervention. On day 29 post-modeling, all rats underwent urodynamic assessments, followed by hematoxylin and eosin (HE) staining, tandem mass tag (TMT) proteomics, and Western blot (WB) analysis of detrusor and bladder neck tissues. Differentially expressed proteins (DEPs) were defined as proteins with P < 0.05, unique peptides ≥ 2, and fold change > 1.2 or < 0.83. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using KOBAS 3.0 (P < 0.01), and protein-protein interaction (PPI) networks were analyzed using Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) 11.5 and Cytoscape 3.9.1. Results: Compared with sham group, DBND group showed significantly elevated leak point pressure (LPP) and maximum cystometric capacity (MCC) (both P < 0.01). EA treatment significantly reduced both LPP and MCC compared with DBND group (P < 0.01 and P < 0.05, respectively). HE staining revealed that EA reduced detrusor fibrosis and improved bladder neck inflammation. TMT proteomics identified 30 overlapping DEPs in detrusor and 59 overlapping DEPs in bladder neck when comparing DBND + EA/DBND groups with sham group. In detrusor tissue, KEGG analysis revealed 10 significantly enriched pathways (P < 0.01), including mitogen-activated protein kinase (MAPK) signaling pathway. PPI analysis showed 22 of 30 DEPs were interconnected. In bladder neck tissue, 14 pathways were significantly enriched (P < 0.01), including relaxin signaling pathway, with 51 of 59 DEPs showing interconnections. Both TMT and WB validations demonstrated that compared with sham controls, DBND rats exhibited upregulated collagen type IV alpha 2 chain (Col4a2) and downregulated guanine nucleotide-binding protein G(z) subunit alpha (Gnaz) in detrusor tissue, while EA treatment normalized both proteins (both P < 0.05). In bladder neck tissue, DBND rats showed decreased expression of smoothelin (Smtn) and calcium-activated potassium channel subunit beta-1 (Kcnmb1) compared with sham controls (both P < 0.01), which were both upregulated following EA treatment (P < 0.01 and P < 0.05, respectively). Conclusion EA restores detrusor-bladder neck coordination in DBND through dual-target mechanisms. In detrusor tissue, EA modulates contraction via extracellular matrix remodeling, cyclic adenosine monophosphate (cAMP) signaling pathway regulation, and enhanced adenosine triphosphate (ATP) biosynthesis mediated by neurotransmitters. In bladder neck tissue, EA promotes relaxation by maintaining contractile phenotypes, reducing fibrosis, suppressing smooth muscle excitation, and regulating presynaptic neurotransmitter release. These findings provide mechanistic insights into EA's therapeutic role in managing DBND.

Objective:To identify core genes with co-expression trends in the suprachiasmatic nucleus (SCN) and lateral habenula (LHb) through miRNA sequencing, and to explore the molecular mechanisms underlying the onset and progress of sleep disturbances in depression.Methods:A rat model of depression with sleep disturbances was established. Behavioral assessments were conducted using the pentobarbital-induced sleep test, sucrose water consumption test, and open field test. Differentially expressed genes (DEGs) in the SCN and LHb of model rats were analyzed using biological replicates with the limma package and DESeq2 software. Gene ontology (GO), KEGG, Reactome pathway-gene set enrichment analysis (GSEA) were performed on the DEGs.Co-expression networks and protein-protein interaction (PPI) networks were constructed using R and Cytoscape software for further screening of genes with potential regulatory roles.Results:After modeling, the sleep latency in the model group ((679.88±350.05) s) was longer than that in the normal group ((316.25±87.35) s) ( t=-2.851, P=0.022), while the sleep duration in the model group ((33.13±8.41) min) was shorter than that in the normal group ((69.72±25.11) min) ( t=3.907, P=0.004). Additionally, the model group exhibited lower sucrose water consumption, horizontal movement counts, and vertical movement counts compared to the normal group (all P<0.05). Differential expression gene analysis showed, in the SCN, 379 genes were upregulated and 132 genes were downregulated, with the top five most significantly differentially expressed genes being Sec61g, Cox6b1, Ephx2, Thrsp, and Pak2. In the LHb, 1 657 genes were upregulated and 1 737 were downregulated, with the top five most significantly differentially expressed genes being Mrpl32, Mrpl20, Mrpl27, Celsr2, and Uba52. Biological pathways enriched in the SCN were mainly associated with the positive regulation of responses to external and biological stimuli. In contrast, pathways enriched in the LHb were related to neurotransmitter level regulation, developmental growth, neurotransmitter transport, and processes such as learning and memory. Co-expression network analysis identified 864 interactions involving 100 nodes in the SCN and 524 interactions involving 99 nodes in the LHb. Conclusion:The biological functions and signaling pathways of the suprachiasmatic nucleus and lateral habenula are different in the occurrence and development of depression with sleep disturbance, and the significantly differentially expressed genes may play an important role.

Objective:To identify core genes with co-expression trends in the suprachiasmatic nucleus (SCN) and lateral habenula (LHb) through miRNA sequencing, and to explore the molecular mechanisms underlying the onset and progress of sleep disturbances in depression.Methods:A rat model of depression with sleep disturbances was established. Behavioral assessments were conducted using the pentobarbital-induced sleep test, sucrose water consumption test, and open field test. Differentially expressed genes (DEGs) in the SCN and LHb of model rats were analyzed using biological replicates with the limma package and DESeq2 software. Gene ontology (GO), KEGG, Reactome pathway-gene set enrichment analysis (GSEA) were performed on the DEGs.Co-expression networks and protein-protein interaction (PPI) networks were constructed using R and Cytoscape software for further screening of genes with potential regulatory roles.Results:After modeling, the sleep latency in the model group ((679.88±350.05) s) was longer than that in the normal group ((316.25±87.35) s) ( t=-2.851, P=0.022), while the sleep duration in the model group ((33.13±8.41) min) was shorter than that in the normal group ((69.72±25.11) min) ( t=3.907, P=0.004). Additionally, the model group exhibited lower sucrose water consumption, horizontal movement counts, and vertical movement counts compared to the normal group (all P<0.05). Differential expression gene analysis showed, in the SCN, 379 genes were upregulated and 132 genes were downregulated, with the top five most significantly differentially expressed genes being Sec61g, Cox6b1, Ephx2, Thrsp, and Pak2. In the LHb, 1 657 genes were upregulated and 1 737 were downregulated, with the top five most significantly differentially expressed genes being Mrpl32, Mrpl20, Mrpl27, Celsr2, and Uba52. Biological pathways enriched in the SCN were mainly associated with the positive regulation of responses to external and biological stimuli. In contrast, pathways enriched in the LHb were related to neurotransmitter level regulation, developmental growth, neurotransmitter transport, and processes such as learning and memory. Co-expression network analysis identified 864 interactions involving 100 nodes in the SCN and 524 interactions involving 99 nodes in the LHb. Conclusion:The biological functions and signaling pathways of the suprachiasmatic nucleus and lateral habenula are different in the occurrence and development of depression with sleep disturbance, and the significantly differentially expressed genes may play an important role.

Objective:To observe any effect of exercise preconditioning on the levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in the brain tissue of rats after induced cerebral ischemia and reperfusion, and how it might promote angiogenesis.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into a sham-operation group, a model group and an exercise preconditioning group, each of 12. After adaptive running training for 3 days, the exercise preconditioning group ran daily for 30 minutes at 15m/min for 14 days, while the other two groups did not exercise. Middle cerebral artery occlusion and reperfusion were then induced in the model and exercise preconditioning groups using the modified Zea-Longa suture method. Rats in the sham-operation group were only cut open to expose the right carotid artery. Right after the modeling, and again 24 hours later neurological deficit was evaluated using the Zea-Longa score and modified neurological severity scoring (mNSS). Infarct sizes were measured using 2, 3, 5-triphenyl tetrazolium chloride staining. Any morphological changes were noted using hematoxylin and eosin (HE) staining, and the expression of CD31 protein, hypoxia-inducible factor-1α and vascular endothelial growth factor in the ischemic cerebral cortex were quantified immunohistochemically.Results:Right after the modelling, compared with the sham-operation group, the average Zea-Longa scores of the model and exercise groups had increased significantly, but were not significantly different from each other. Twenty-four hours later the average Zea-Longa score, mNSS score and relative cerebral infarction area of the model group had increased significantly compared with the sham-operation group, while the exercise preconditioning group′s averages had decreased significantly. The HE staining showed that compared with the sham-operation group, pathological changes such as loose tissue, reduced number of nerve cells, nucleolysis, and vacuolization of the cerebral cortex on the ischemic side were found in the model group. Compared with the model group, the pathological changes in the exercise preconditioning group were less serious. The levels of CD31 protein, HIF-1α and VEGF in the ischemic cerebral cortexes of the model group had by then increased significantly. But compared with the model group, those levels had increased more in the exercise preconditioning group.Conclusion:Exercise preconditioning can effectively promote angiogenesis after cerebral ischemia and reduce chronic injury. That may be related to the activation of the HIF-1α and/or VEGF signaling pathways.