南方医科大学学报杂志最新文献

Objectives: To investigate the molecular mechanism of helicid for improving depressive-like behaviors in rats exposed to chronic unpredictable mild stress (CUMS).

Methods: SD rats were randomly divided into normal control group (n=20) and CUMS group (n=70) to receive no stimulation and mild unpredictable stress for 6 weeks, respectively. After successful modeling, CUMS rats were further divided into 7 subgroups for intracerebroventricular injection with saline, adeno-associated virus (AAV) vector, or AAV carrying si-NCALD (NCALD silencing experiment, n=10); or intracerebroventricular injection with saline, saline with daily helicid gavage, AAV vector with helicid gavage, or NCALD-overexpressing AAV with helicid gavage (NCALD overexpresison experiment, n=10). The depressive state of the rats was evaluated by assessing changes in body weight, sucrose preference, and open field test. The expressions of NCALD, sGCα1, sGCβ1, PKG1/2, and cleaved-caspase 3 in the hippocampus of the rats were detected by Western blotting, and hippocampal cGMP level was determined with ELISA.

Results: Compared with the normal control rats, CUMS rats showed significantly increased hippocampal expressions of NCALD and cleaved caspase-3 and abnormal activation of the sGC/cGMP/PKG pathway. Silencing NCALD by intracerebroventricular injection of AAV-si-NCALD significantly reduced cleaved caspase-3 and inhibited sGC/cGMP/PKG pathway activation in the hippocampus, and improved depressive-like behaviors of the rats. Helicid treatment produced similar effects, but its effect was abolished by intracerebroventricular injection of NCALD-overex-pressing AAV.

Conclusions: Helicid relieves depressive-like behaviors in CUMS rats by downregulating NCALD, inhibiting abnormal sGC/cGMP/PKG activation, and reducing hippocampal apoptosis.

Objectives: To identify the differentially expressed genes in obese asthma versus non-obese asthma and evaluate the effect of acupuncture on chronic airway inflammation in obese mice with asthma.

Methods: The key genes of obesity-related asthma were screened using GSE110551 dataset from the GEO database, and the characteristic genes were selected from the genes with the highest correlation with T cells using Lasso regression and SVM feature selection algorithms. Fifty C57BL/6J mice (5-6 weeks old) were randomized equally into 5 groups, including a normal feeding (control) group, a high-fat feeding group, and 3 high-fat feeding and ovalbumin sensitization groups with intraperitoneal injections with saline or dexamethasone (DEX), or treated with acupuncture. Western blotting, qPCR, and flow cytometry were used to analyze the changes in the expressions of the key genes and inflammation in the airway of the mice.

Results: FAM126B and VNN1 were identified as the characteristic genes in obesity-related asthma for subsequent analysis. The mice with high-fat feeding and ovalbumin sensitization showed the highest expression levels of Vnn1 and FAM126B among the 5 groups, with also significantly decreased Treg cell percentage and obvious inflammatory cell infiltration in the lungs. Treatment with DEX and acupuncture both significantly decreased the number of infiltrating inflammatory cells and increased the percentage of Treg cells in airway of the mouse models of obesity-related asthma. HIF-1α was identified as a key regulatory factor for asthmatic inflammation, and its expression level was significantly increased in the asthmatic mouse models but obviously lowered after acupuncture treatment or dexamethasone therapy.

Conclusions: Vnn1 and FAM126B may serve as the key therapeutic targets for treatment of obese asthma patients. Acupuncture treatment may downregulate airway HIF-1α by reducing the expressions of Vnn1 and FAM126B and increasing the number of Treg cells.

Objectives: We propose an efficient deep learning model to improve the classification accuracy in automatic classification tasks of 12-lead electrocardiogram (ECG) signals.

Methods: We designed a new ResLSTM-TemporalSE network architecture by incorporating a multi-layer Residual Long Short-Term Memory (ResLSTM) structure and introducing skip connections between LSTM layers to establish residual learning pathways for the temporal features. A temporal attention mechanism was integrated into the traditional Squeeze-and-Excitation (SE) module to enhance channel-wise feature representation while capturing long-term temporal dependencies within ECG signals, thereby an efficient hierarchical feature extraction framework was constructed. The model was validated using the public CPSC2018 dataset and a private clinical dataset from the Seventh Affiliated Hospital of Southern Medical University.

Results: The experimental results demonstrated that the model achieved a classification accuracy of 99.70% on the CPSC2018 test set, with precision, recall, and F1-score values of 0.9966, 0.9370, and 0.9653, respectively. On the private clinical dataset, it attained an accuracy of 82.77%, with precision, recall, and F1-score values of 0.6811, 0.8961, and 0.7723. Ablation studies confirmed the significant contributions of both the residual connections and the temporal attention module to model performance.

Conclusions: The ResLSTM-TemporalSE model effectively integrates spatiotemporal features of the ECG signals and demonstrates superior classification performance on the CPSC2018 benchmark while maintaining strong generalization capabilities in real-world clinical settings. This framework provides a robust solution for automated ECG analysis and holds significant promise for clinical applications.

Objectives: To explore the therapeutic mechanism of Fuzheng Xiaoyan (FZXY) Granules for relieving cancer-related fatigue (CRF) during chemotherapy in breast cancer patients based on the traditional Chinese medicine (TCM) theory of "Fuzheng Quxie" (supporting healthy qi and eliminating pathogens).

Methods: Ninety CRF patients with breast cancer and Zhengxu Duyu syndrome were randomized equally into control group with chemotherapy and symptomatic treatment and study group with additional treatment with FZXY Granules, and their Piper Fatigue Scale (PFS), Karnofsky Performance Status (KPS), and TCM syndrome scores were compared. Network pharmacology analysis was used to identify the active components in FZXY Granules, the drug targets, and disease-related targets. Protein-protein interaction (PPI) network was constructed followed by enrichment analysis. Molecular docking study was conducted to explore the interactions between quercetin and the core targets. In a CRF mouse model bearing breast cancer xenograft, the effects of saline and FZXY Granule gavage were observed by assessing motor function, expressions of AKT1, p-AKT1, BCL-2, and BAD in the gastrocnemius muscle, and serum levels of IL-6 and IL-1β.

Results: The patients receiving FZXY Granules treatment showed significantly improved PFS, KPS, and TCM syndrome scores compared with the baseline levels and those in the control group (P<0.05). Fifty-seven overlapping drug-disease targets were screened, and 5 core targets were identified. Quercetin exhibited strong binding to AKT1 and acted likely via the apoptosis pathways. In the CRF mouse models, FZXY Granules obviously improved motor function of the mice, reversed abnormal apoptosis-related protein expressions in the gastrocnemius muscle, and reduced serum IL-6 and IL-1β levels.

Conclusions: FZXY Granules alleviate CRF and improve TCM symptoms and quality of life of breast cancer patients during chemotherapy possibly by suppressing skeletal muscle cell apoptosis via regulating the AKT1/BAD/BCL-2 pathway and reducing IL-6 and IL-1β levels.

Objectives: To investigate the effect of overwork on myocardial energy metabolism in mice.

Methods: Thirty-two C57BL/6J mice were randomized equally into a control group and 3 overwork groups with overwork for 2, 4, and 6 weeks (W2, W4, and W6 groups, respectively). The mice in overwork groups were subjected to daily forced water standing and restraint. The changes in body weight and general condition of the mice were observed weekly. After successful modeling, the mice were examined for changes in echocardiography, blood glucose/lipid profiles, myocardial pathologies, myocardial TG and ATP levels, and expressions in CD36, GLUT1, CPT1B, PPARα, PFKM, and PKM2 using immunohistochemistry, RT-qPCR or Western blotting.

Results: The mice with prolonged overwork exhibited reduced activity with hair loss, dull fur, and slowed body weight gain without significant changes in cardiac index or function. Blood glucose levels increased significantly in W2 and W4 groups but decreased in W6 group. Serum TG level increased significantly while TC, HDL, and LDL decreased in W4 and W6 groups. HE staining revealed myocardial swelling, disorganization, and vacuolation in the mouse models. Myocardial TG was elevated in W4 and W6 groups and ATP level decreased in W6 group. The mRNA and protein expressions of CPT1B and PPARα were downregulated in W4 and W6 group, and CD36 expression increased significantly in W4 group. GLUT1 and PFKM/PKM2 expressions decreased obviously in W2 group but increased in W4 and W6 group compared with that in W2 group.

Conclusions: Short-term overwork causes elevation of blood glucose and suppresses glycolysis in mice, while prolonged overwork reduces glucose, increases TG, impairs fatty acid oxidation, and limits glycolytic compensation to eventually result in myocardial damage, lipid accumulation, and ATP deficiency.

Objectives: To investigate the role of Cav1 gene in regulating mitochondrial function and mRNA m⁶A modification and expressions of the key genes in mouse hepatocytes.

Methods: In mouse hepatocyte AML12 cells, the effects of lentivirus-mediated Cav1 knockdown and transfection with a negative control virus on mitochondrial membrane potential and mitochondrial respiratory function were analyzed using the TMRE fluorescent probe and the Seahorse extracellular flux analysis system. Methylated RNA immunoprecipitation (MeRIP) combined with m⁶A microarray was utilized to evaluate the changes in mRNA m⁶A modification and expression levels, followed by enrichment analysis to identify the functionally relevant genes. The m⁶A modification and expression levels of the mRNAs were validated by qPCR.

Results: Compared with the negative control group, the cells with Cav1 knockdown exhibited significantly reduced mitochondrial membrane potential and respiratory capacity. m⁶A microarray analysis revealed significant changes in m⁶A modification levels (fold change>1.5) of 7814 mRNAs, including 152 upregulated and 7662 downregulated mRNAs. Integrated expression analysis identified 2497 mRNAs showing coordinated changes in m⁶A modification and expression levels. These mRNAs were enriched in the oxidative phosphorylation pathway, with Usp15, Suclg2, and Ppa2 exhibiting the highest percent changes in m⁶A modification. Both microarray and qPCR results showed that the m⁶A modification and expression levels of Usp15, Suclg2 and Ppa2 mRNAs were significantly reduced in cells with Cav1 knockdown compared to the NC group.

Conclusions: Cav1 knockdown induces significant alterations in mRNA m⁶A modification as well as their expression levels. The regulatory effects of Cav1 on mitochondrial function may be mediated by modulation of m⁶A modification of Usp15, Suclg2, and Ppa2 mRNAs.

Objectives: To investigate the clinical significance of abnormal expression of angio-associated migratory cell protein (AAMP) in hepatocellular carcinoma (HCC).

Methods: Bioinformatics analyses were used to analyze AAMP expression level in HCC and its prognosis value. In 60 pairs of HCC and adjacent tissues, AAMP expression was detected immunohistochemically and its correlation with clinicopathological characteristics of the patients was analyzed. In cultured Mahlavu and Huh-7 cells with lentivirus-mediated AAMP knockdown, the changes in cell proliferation, apoptosis, migration and invasion were observed, and their lung metastasis following tail vein injection in nude mice were assessed. In HCC cells with AAMP knockdown, Western blotting, immunofluorescence staining or RT-qPCR were used to examine the changes in expression levels of E-cadherin, N-cadherin, Vimentin and Snail and the effects of MG-132 and CHX on RhoA expression. The correlation between the expressions of AAMP and RhoA in HCC tissues was analyzed by immunohistochemistry.

Results: Bioinformatics analysis showed that AAMP expression was elevated in HCC tissues (P<0.05) in correlation with advanced clinical stage and poor prognosis (P<0.05). Immunohistochemistry results confirmed significant correlations of high AAMP expression with Edmondson-Steiner grade (III+IV), venous infiltration and TNM stage (III+IV) of HCC (P<0.05). In cultured HCC cells, AAMP knockdown did not significantly affect cell proliferation or apoptosis, but obviously suppressed cell migration and invasion in vitro and lung metastasis in nude mice. AAMP knockdown significantly increased E-cadherin expression, decreased N-cadherin, Vimentin and Snail expressions, and reduced RhoA protein levels without obviously affecting RhoA mRNA levels. MG-132 treatment blocked the inhibitory effect of AAMP knockdown on RhoA protein expression. The expressions of AAMP and RhoA showed a significant positive correlation in HCC tissues (P<0.05).

Conclusions: AAMP overexpression is associated with malignant clinical features of HCC and promotes epithelial-mesenchymal transition and metastasis of HCC cells partly by stabilizing RhoA expression.

Objectives: To explore the mechanism of Shenqi Xiezhuo Decoction (SQXZD) for improving renal fibrosis (RF) in rats.

Methods: The chemical components of SQXZD were identified using UPLC-Q Exactive/MS, and component-disease target network and enrichment analyses were conducted to screen the key pathways and targets. In the animal experiment, 49 male SD rats were randomized equally into blank control group, sham operation group, unilateral ureteral obstruction-induced RF model group, losartan treatment (daily dose 4.6 mg/kg) group, and low-, medium-, and high-dose SQXZD (9.7, 19.4, and 38.8 g/kg, respectively) treatment groups. After 14 days' treatment, renal pathologies and collagen deposition of the rats were examined with HE and Masson staining, and serum levels of BUN, Cr, SOD, MDA, GSH-px, IL-6, and TNF-α were detected. Western blotting and qRT-PCR were used to detect renal protein and mRNA expressions of α‑SMA, Col-I, NAKED2, Rap1, B-raf, Raf-1, MEK3/6, p38MAPK, MEK, ERK1/2, p-ERK1/2, FoxO3a, p-FoxO3a, and MnSOD.

Results: A total of 263 chemical components were identified in SQXZD. Network pharmacology revealed 170 intersecting targets between the components and RF enriched in the MAPK, Rap1, and FoxO pathways. The rat models of RF showed abnormal renal structural changes, increased fibrosis area, elevated serum BUN, Cr, MDA, IL-6, and TNF-α levels, reduced SOD and GSH-px levels, upregulated renal expressions of α‑SMA, Col-I, NAKED2, Rap1, B-raf, MEK, ERK1/2, p-ERK1/2, MEK3/6, and p38MAPK, and downregulated Raf-1, FoxO3a, p-FoxO3a, and MnSOD expressions. Treatment with losartan and SQXZD (especially at the medium dose) obviously lessened renal pathologies, improved renal functions, alleviated oxidative stress and inflammation, and ameliorated abnormal changes in the Rap1/MAPK/FoxO3a signaling pathway in the rat models.

Conclusions: SQXZD alleviates RF and improves renal function in rats possibly by ameliorating renal oxidative stress and inflammation via regulating the Rap1/MAPK/FoxO3a signaling pathway.

Objectives: To investigate the inhibitory effect of polyphyllin VII (PP7) on osteosarcoma xenograft growth in mice and explore the underlying molecular mechanism.

Methods: Ultra‑performance liquid chromatography‑tandem mass spectrometry was used to analyze the main active components of Paris polyphylla. Six nude mice bearing patient‑derived xenograft (PDX) were randomized into two groups for treatment with 2 mg/kg PP7 gavage or saline every other day for 28 days, and the changes in tumor volume and mass were measured. In cultured 143B and HOS cells, the effect of PP7 treatment (0, 1.25, 2.5, 5, and 10 μmol/L) on cell proliferation was assessed with CCK‑8 assay, and Transwell assays were employed to examine the changes in cell migration and invasion. The target of PP7 was predicted by integrated analyses with single‑cell RNA sequencing (scRNA‑seq), bulk RNA sequencing (bulk RNA‑seq) and molecular docking and verified using Western blotting. In osteosarcoma cells transfected with SOHLH1 siRNAs or a negative control sequence, the effects of PP7 treatment (5 μmol/L) on cell migration, invasion, ferroptosis, reactive oxygen species (ROS) production and lipid peroxidation (LPO) were analyzed.

Results: PP7 was identified as one of the major active constituents of Paris polyphylla. In the tumor-bearing mice, PP7 treatment significantly lower the tumor volume and mass. In 143B and HOS cells, PP7 concentration‑dependently inhibited cell proliferation, and at 5 μmol/L, PP7 significantly inhibited cell proliferation, migration and invasion. Multi‑omics analysis identified SOHLH1 as a potential target of PP7, and Western blotting confirmed that PP7 upregulated SOHLH1 expressions at both the mRNA and protein levels. SOHLH1 silencing obviously attenuated the inhibitory effects of PP7 on cell migration and invasion and reduced PP7‑induced ferroptosis.

Conclusions: PP7 suppresses osteosarcoma xenograft growth in mice by inducing ferroptosis via upregulating SOHLH1 expression.

Objectives: To investigate the mechanism by which Morinda officinalis polysaccharide (MOP) delays osteoarthritis chondrocyte degeneration.

Methods: In primary cultures of chondrocytes from 4-week-old C57BL/6 mice, the effects of IL-1β and MOP treatment at different concentrations on cell viability were assessed with CCK-8 assay. The treated cells were examined for protein expressions of PKM2, caspase-1, and GSDMD using Western blotting and for XIST expression using fluorescence in situ hybridization (FISH). In IL-1β-induced mouse chondrocytes, the effects of MOP, transfection for XIST overexpression or knockdown, and MOP treatment after the transfection were tested by detecting mRNA levels of GluT1, HK2, PKM2, LDHA, PFKFB3, NLRP3, caspase-1, and GSDMD; flow cytometry, Western blotting, and toluidine blue staining were used to analyze chondrocyte apoptosis, expressions of glycolysis and pyroptosis regulators, and glycosaminoglycan expression.

Results: The second-passage chondrocytes showed good viability and positive collagen II staining. IL-1β induction caused degenerative morphological changes of the cells, decreased collagen II expression, and upregulated cellular expressions of PKM2, caspase-1, and GSDMD proteins. MOP treatment (especially at 4 mg/mL) significantly enhanced cell viability and reduced HK2, PKM2, caspase-1 and GSDMD expressions in IL-1β‑induced mouse chondrocytes. XIST was localized predominantly in the nuclei of the chondrocytes, and its expression increased significantly in IL-1β‑treated cells, and was attenuated by MOP treatment. XIST overexpression synergized with IL-1β to upregulate mRNA and protein expressions of glycolysis- and pyroptosis-related factors in the chondrocytes, and such effects were obviously attenuated by MOP. Conversely, XIST knockdown significantly inhibited chondrocyte apoptosis and glycosaminoglycan expression, and down-regulated glycolysis- and pyroptosis-related proteins. MOP treatment exhibited similar protective effects to XIST knockdown, and their combination significantly augmented these protective effects.

Conclusions: MOP mitigates IL-1β‑induced mouse chondrocyte degeneration by modulating glycolysis and pyroptosis via targeting XIST.