1.Polygonatum Sibiricum Polysaccharides Improve Colonic Injury in a Mouse Model of Chronic Obstructive Pulmonary Disease by Regulating Bile Acid Metabolism in the Colon
Wanrong LI ; Mengting TAO ; Yuanfeng ZOU ; Dan HE ; Nengyuan TANG ; Xin TAN ; Lixia LI ; Dandan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(3):431-443
ObjectiveTo investigate the effect and mechanism of Polygonatum neutral polysaccharides from sibiricum (PSP-NP) on colon injury in mice with chronic obstructive pulmonary disease (COPD). MethodsMale C57BL/6J mice were randomly divided into a control group, a COPD model group, and a PSP-NP group. The COPD model was established using smoke exposure combined with intranasal LPS administration. The PSP-NP group was simultaneously treated daily with 200 mg/kg of PSP-NP via intragastric gavage, while the other groups received an equal volume of saline. HE staining was used to observe the pathological changes in the colon. ELISA was employed to detect the levels of LPS in serum and the expressions of ZO-1, Occludin, IL-6, and TNF-α in colon tissue. UPLC-MS was used to detect the types and contents of bile acids in colonic content, and to screen for differential bile acids. Differential microbial flora were identified using 16S rRNA gene sequencing, and correlation analysis was conducted with differential bile acids. PSP-NP was combined with the differential bile acids cholic acid (CA), and deoxycholic acid (DCA) in vitro to analyze the binding capacity of PSP-NP for CA and DCA. PSP-NP was applied to NCM460 normal colonic epithelial cells cultured in CA and DCA. Cell migration ability was assessed using the scratch assay, and the mRNA expression levels of inflammatory cytokines TNF-α, IL-6, and NF-κB were measured by RT-qPCR. ResultsPSP-NP effectively improved colonic damage in COPD model mice, enhanced mechanical barrier function, alleviated inflammatory response, and regulated abnormal changes in colonic flora and bile acid metabolism. Correlation analysis further revealed that PSP-NP regulated colonic bile acid metabolism and reduced the redundancy of secondary bile acids by increasing the relative abundance of Bacteroidota, Verrucomicrobiota, Bacteroides, and Akkermansia, while decreasing the relative abundance of Lactobacillus and Bifidobacterium. Notably, in vitro binding assays demonstrated that PSP-NP bound to differential bile acids DCA and CA, with the strongest binding capacity for DCA at 58.2%. In cellular functional studies, DCA inhibited the migration ability of colonic epithelial cells NCM460 and significantly increased the relative mRNA expression levels of inflammatory factors TNF-α, IL-6, and NF-κB. Importantly, co-treatment with PSP-NP significantly ameliorated the impact of DCA on NCM460 cells. ConclusionsPSP-NP may significantly improve colonic damage in COPD model mice. The mechanism may involve the regulation of colonic bile acid metabolism and bile acid profiles through both microbial modulation and direct binding, thereby reducing the damage caused by secondary bile acids such as DCA to colonic epithelial cells.
2.Houshihei San Repairs Skeletal Muscle Injury After Ischaemic Stroke by Regulating Ferroptosis Pathway
Hu QI ; Dan TIAN ; Xiongwei ZHANG ; Zeyang ZHANG ; Yuanlin GAO ; Yanning JIANG ; Xinran MIN ; Jiamin ZOU ; Jiuseng ZENG ; Nan ZENG ; Ruocong YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):1-11
ObjectiveTo investigate the pharmacodynamic effects of Houshihei San (HSHS) recorded with the effects of treating wind and limb heaviness on muscle tissue injury after middle cerebral artery occlusion (MCAO) in rats through the ferroptosis pathway. MethodsThirty SD male rats were selected and randomly grouped as follows: sham, MCAO, deferoxamine mesylate, high-dose HSHS (HSHS-H, 0.54 g·kg-1), and low-dose HSHS (HSHS-L, 0.27 g·kg-1), with 6 rats in each group. A laser scattering system was used to evaluate the stability of the MCAO model, and rats were administrated with corresponding agents by gavage for 7 days. During the administration period, behavioral, imaging and other methods were used to systematically evaluate the skeletal muscle tissue injury after MCAO and the therapeutic effect in each administration group. Hematoxylin-eosin staining was employed to evaluate the cross-section of muscle cells. Subsequently, immunohistochemistry was used to detect tumor suppressor p53 and glutathione peroxidase 4 (GPX4) in the soleus tissue. Western blot was employed to determine the protein levels of p53, GPX4, myogenic differentiation 1 (MyoD1), nuclear factor E2-related factor 2 (Nrf2), Myostatin, solute carrier family 7 member 11 (SLC7A11), muscle ring-finger protein-1 (MuRF1), and muscle atrophy F-box protein (MAFbx) to verify the therapeutic effect in each group. ResultsCompared with the MCAO group, HSHS enhanced the locomotor ability and promoted muscle regeneration, which suggested that the pharmacological effects of HSHS were related to the inhibition of muscle tissue ferroptosis to reduce the expression of muscle atrophy factors. Behavioral and imaging results suggested that compared with the MCAO group, HSHS ameliorated neurological impairments in rats on day 7 (P<0.01), enhanced 5-min locomotor distance and postural control (P<0.01), strengthened grasping power and promoted muscle growth (P<0.01), stabilized skeletal muscle length and weight (P<0.01), and increased the cross-section of muscle cells (P<0.01). Compared with the MCAO group, HSHS promoted the increases in glutathione and superoxide dismutase content and inhibited the increase in malondialdehyde content (P<0.05,P<0.01). Ferroptosis pathway-related assays suggested that HSHS reduced the p53-positive cells and increased the GPX4-positive cells (P<0.01). HSHS ameliorated muscle function decline after stroke by promoting the expression of GPX4, Nrf2, SLC7A11, and MyoD1 and inhibiting the expression of p53, Myostatin, MurRF1, and MAFbx to reduce ferroptosis in the muscle (P<0.01). ConclusionHSHS, prepared with reference to the method in the Synopsis of Golden Chamber, can simultaneously reduce the myolysis and increase the protein synthesis in the skeletal muscle tissue after ischemic stroke by regulating the ferroptosis pathway.
4.Visualization analysis on research hotspots and trends of Yupingfeng Powder from 2000 to 2023
Ying ZOU ; Nijina LI ; Suhui TAN ; Yuting HUANG ; Dan ZHANG ; Hao ZHANG
International Journal of Traditional Chinese Medicine 2025;47(1):96-101
Objective:To analyze the research hotspots and trends of Yupingfeng Powder from 2000 to 2023; To provide reference for related research.Methods:The research literature related to Yupingfeng Powder was retrieved from CNKI, VIP, Wanfang Data and SinoMed from January 1, 2000 to December 20, 2023. Excel 2022 software was used to analyze the annual number of publications. CiteSpace 6.1.R6 software was used for visualization analysis on the authors, research institutions and key words.Results:A total of 2 522 articles were included, and the number of published articles showed a fluctuating growth trend in recent years. The journal with the largest number of articles was New Chinese Medicine (55 articles). The authors with the most publications were Hong Min from Nanjing University of Chinese Medicine (9 articles) and Zhang Zhonglin from Chengdu Medical College (9 articles). Yunnan University of Chinese Medicine (13 articles) had the largest number of publications. High-frequency keywords included allergic rhinitis, children, urticaria, recurrent respiratory tract infection, etc. The clustering analysis generated 11 labels. Conclusions:The research hotspots of Yupingfeng Powder focus on clinical application, integrated traditional Chinese and Western medicine therapy, syndrome type research and pharmacological research. Predicting potential drug targets with the help of network pharmacology and molecular docking technology and exploring its mechanism are the research trends in this field.
5.Visualization analysis on research hotspots and trends of Banxia Houpu Decoction based on CiteSpace
Nijina LI ; Ying ZOU ; Hao ZHANG ; Yamin LI ; Dan ZHANG
International Journal of Traditional Chinese Medicine 2025;47(8):1141-1146
Objective:To analyze the research status, hotspots, and trends of Banxia Houpu Decoction.Methods:Research literature about Banxia Houpu Decoction was retrieved from the CNKI, VIP, Wanfang Data, and SinoMed from January 1, 2000 to June 30, 2024. NoteExpress 3.6.0.9220 software was used to merge and eliminate duplicates, and CiteSpace 6.1.R6 software was used to draw and interpret the annual publication volume, authors, and keywords.Results:A total of 716 articles were included in this study, and the number of articles increased first and then decreased in recent years. The journal with the most published articles was Journal of Practical Traditional Chinese Medicine (23 articles); the author with the largest number of articles was Kong Lingdong from Nanjing University (7 articles); high-frequency keywords included "globus hystericus", "cough" and "chronic pharyngitis". A total of 11 labels were generated by clustering analysis, which could be summarized into two topics: clinical application (chronic pharyngitis, gastroesophageal reflux disease, aspiration pneumonia, depression, etc.) and literature type (clinical observation, experimental research). Conclusions:The research hotspots of Banxia Houpu Decoction are focused on clinical application and compatibility research. Using network pharmacology to predict potential drug targets and explore their mechanisms, conducting experimental research to explore pharmacological effects and clinical applications for treating diseases is a research trend in this field.
6.Inhibitory effect of β-elemonic acid on proliferation and invasion of colon cancer cell lines
Ting SUN ; Teng ZOU ; Yisong YANG ; Shuangping LIU ; Xin REN ; Dan WANG
Basic & Clinical Medicine 2025;45(6):748-754
Objective To investigate the inhibitory effect of β-elemonic acid(β-EA)on the proliferation and inva-sion of colon cancer cells and the underlying mechanisms through PI3K/AKT/mTOR signaling pathway.Methods The effects of β-EA on colon cancer cell proliferation were evaluated using the MTT assay and colony formation as-say.Transwell invasion assay were used to assess the impact of β-EA on invasion.Western blot analysis was con-ducted to detect changes in PI3K/AKT/mTOR pathway proteins after treatment.Results MTT assay showed thatβ-EA effectively inhibited the proliferation of colon cancer HCT8 and HCT116 cells in a dose-dependent manner.The colony formation assay confirmed its inhibitory effect on cell proliferation.Transwell invasion assays demonstra-ted that β-elemonic acid reduced the invasion abilities of the cells.Western blot analysis revealed increased expression of apoptosis-related proteins cleaved-caspase 3,cleaved-caspase 9,and Bax,while Bcl-2 expression was decreased.Invasion-related proteins vimentin,snail,MMP2,and MMP9 were downregulated after treatment.Addi-tionally,β-EAA reduced the levels of p-PI3K,p-Akt,and p-mTOR,and these reductions were more pronounced af-ter the addition of the PI3K inhibitor LY294002.Conclusions β-EA may inhibit proliferation and invasion in colon cancer cell lines HCT8 and HCT116 through PI3K/AKT/mTOR signaling pathway,and potentially be transformed as a novel therapeutic agent for colon cancer.
7.Inhibition of Pannus Formation by Wutoutang in AIA Rats with Wind-cold-dampness Bi Syndrome Via Down-regulating HIF-1α/VEGFA/Ang Signaling Pathway
Dan OUYANG ; Xin LI ; Gangxiang ZHAN ; Zheng CHEN ; Yuanzhi MA ; Jie ZOU ; Jianzhong CAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):10-18
ObjectiveThis study aims to investigate the inhibitory effect of Wutoutang on pannus formation in adjuvant-induced arthritis (AIA) rats with wind-cold-dampness Bi syndrome and its potential mechanism. MethodA total of 40 male SD specific pathogen-free (SPF) rats were selected and divided into blank group, wind-cold-dampness Bi syndrome group [Complete Freund's Adjuvant (CFA), 200 μg], Wutoutang group (15 g·kg-1·d-1), and indometacin group (10 mg·kg-1) according to random number table method. Except for the blank group, the other groups were given wind-cold-dampness stimulation before the CFA injection. After the rats were administered for 30 days, the basic conditions, onset time, arthritis index score, and foot swelling volume of AIA rats with wind-cold-dampness Bi syndrome were observed. Finally, peripheral arterial blood, ankle joint, and synovial tissue were taken. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor A (VEGFA) protein content, and rheumatism, including anti-O (ASO), C-reactive protein (CRP), and rheumatoid factor (RF). Hematoxylin-eosin (HE) staining revealed the changes in joint histomorphology. Immunohistochemistry was used to detect the expression of HIF-1α and VEGFA, two important proteins in the ankle pathway. Quantitative real-time polymerase chain reaction (Real-time PCR) was used to reveal mRNA levels of HIF-1α, VEGFA, angiopoietin-1 (Ang-1), and angiopoietin-2 (Ang-2) in rat synovial tissue. ResultThe foot swelling volume and arthritis score of AIA rats with wind-cold-dampness Bi syndrome were substantially higher (P<0.01) compared with the blank group. Serum CRP, RF, and ASO levels were considerably elevated (P<0.01). HE staining showed obvious hyperplasia of ankle synovium and synovial inflammation, angiogenesis and pannus formation, and aggravated bone destruction, indicating successful modeling. After the intervention of Wutoutang, the onset time was delayed (P<0.01). Foot swelling volume and arthritis score were decreased (P<0.01). Serum CRP, RF, and ASO levels were significantly decreased (P<0.01). The inflammatory hyperplasia of synovial tissue, angiogenesis and pannus formation, and bone destruction were alleviated. The mRNA levels of HIF-1α, VEGFA, Ang-1, and Ang-2 in the synovial membrane were significantly decreased (P<0.05, P<0.01). The expressions of HIF-1α and VEGFA in serum and ankle joints were decreased (P<0.01). In the indomethacin group, the onset time of the disease was delayed (P<0.01). Foot swelling volume and arthritis score were decreased (P<0.01). Serum CRP, RF, and ASO levels were significantly decreased (P<0.01). HIF-1α/VEGFA/Ang signaling pathway was activated, and pathological tissue injury was improved. ConclusionWutoutang can delay the onset time of AIA rats with wind-cold-dampness Bi syndrome, reduce foot swelling volume, arthritis score, rheumatic activity, and improve joint histopathology. It can inhibit pannus formation, and its mechanism may be related to down-regulating the expression of the HIF-1α/VEGFA/Ang pathway.
8.Association of different sleep characteristics and cardiometabolic risk in college students
Chinese Journal of School Health 2024;45(1):25-29
Objective:
To describe the association of different sleep characteristics and cardiometabolic risk among college students, so as to provide reference for health promotion of college students.
Methods:
By random cluster sampling method, a questionnaire survey and physical examination including blood pressure, waist circumference and blood lipid indicators, which were conducted in April and May of 2019 among a total of 1 179 college students from the first grade in two universities in Hefei City of Anhui Province and Shangrao City of Jiangxi Province. A total of 729 college students with valid questionnaires were included into analysis. The Pittsburgh Sleep Quality Index (PSQI) and Insomnia Severity Index (ISI) were used to investigate sleep behavior, and the Morning And Evening Questionnaire-5 (MEQ-5) was used to investigate sleep characteristics. The cardiometabolic risk score was derived using the sum of the standardized sex specific Z scores of waist circumference, mean arterial pressure, HDL cholesterol (multiplied by -1), triglycerides, and insulin resistance index. The rank sum tests were used to compare differences in cardiometabolic risk scores across demographic characteristics. Generalized linear models were used to compare the association of different sleep characteristics with cardiometabolic risk scores among college students.
Results:
The average cardiovascular metabolic risk score of college students was -0.32(-2.03, 1.58). There were statistically significant differences in cardiovascular metabolic risk scores among college students in variables such as smoking, health status, and physical activity levels ( t/F=-3.41, 12.88, 51.07, P <0.01). The results of the generalized linear model showed that nighttime preference ( B=1.89, 95%CI =1.02-3.49), insomnia symptoms ( B=3.25, 95%CI =1.79-5.90), and short or long sleep duration ( B=1.92, 95%CI =1.21-3.05) were positively correlated with the cardiovascular metabolic risk score of college students ( P <0.05).
Conclusions
Poor sleep patterns among college students are positively correlated with the risk of cardiovascular metabolism. The sleep behavior of college students should be actively changed to reduce the risk of cardiovascular disease.
9.The Meta-analysis of efficacy and safety of edaravone combined with sodium valproate in treatment of post-stroke epilepsy
Dan ZOU ; Xingyong YANG ; Qiaozhi HU
China Pharmacist 2024;27(1):145-154
Objective To systematically evaluate the efficacy and safety of the combination therapy of sodium valproate and edaravone in the treatment of post-stroke epilepsy(PSE).Methods The Cochrane Library,PubMed,Embase,Web of Science,CNKI,Wanfang database,and VIP databases were searched from the time of inception to May 2023.Randomized controlled trials of edaravone plus valproate versus valproate monotherapy for epilepsy were collected,the risk of bias of the included studies was assessed using the RoB 2 tool,and Meta-analyses were performed using RevMan 5.4 software.Results 13 studies with a total of 1 092 patients were included.The results showed that the response rate was significantly higher in the combination group than in the sodium valproate group[RR=0.18,95%CI(0.13 to 0.22),P<0.01].The incidence of adverse reactions was lower[RR=0.73,95%CI(0.48 to 1.13),P=0.16].The seizure frequency[MD=-0.30,95%CI(-0.43,-0.11),P<0.01]and duration of seizures[MD=-0.81,95%CI(-0.89,-0.72),P<0.01]in the combined treatment group were significantly lower than those in the sodium valproate group.The inflammatory factors tumor necrosis factor-α[MD=-8.00,95%CI(-9.15,-6.84),P<0.01],interleukin-2[MD=-10.19,95%CI(-14.61,-5.78),P<0.01],interleukin-8[MD=-5.6,95%CI(-6.48,-4.73),P<0.01]were significantly lower than those in the sodium valproate group.The neuron-specific enolase(NSE)level in patients with epilepsy in the combination treatment group was significantly better than that of the sodium valproate group at 1 month[MD=-4.73,95%CI(-4.99,-4.46),P<0.01],3 months[MD=-2.10,95%CI(-3.26,-0.95),P<0.01],and 6 months after treatment[MD=-1.31,95%CI(-2.35,-0.27),P<0.01].There was no significant difference in NSE levels[MD=0.06,95%CI(-0.07,0.19),P=0.34]after 12 months.Conclusion Edaravone combined with sodium valproate in the treatment of post-stroke epilepsy can improve the epilepsy control rate of patients,shorten the frequency and duration of seizures,reduce the level of inflammatory factors in patients,promote the reduction of NSE levels,and improve their quality of life,which has a certain safety.
10.Mineralization regulation of MAGE-D1 on bone marrowmesenchymal stem cells in knockout mice
Mingjie LU ; Hongyan YUAN ; Dan XU ; Xuelian PENG ; Xuqiang ZOU ; Bo XIE ; Jingwen MAO ; Xiujie WEN
Journal of Army Medical University 2024;46(18):2069-2080
Objective To investigate the effect of melanoma associated antigen D1 (Mage-D1)on mouse femoral bone mass and mineralization ability of mouse bone marrow mesenchymal cells (BMSCs)and its potential molecular mechanism.Methods Female Mage-D1 gene knockout heterozygous mice and male wild-type (WT)mice were subjected as parent mice to breed Mage-D1 gene knockout homozygous (Mage-D1 KO)mice.PCR and agarose gel electrophoresis were used to identify male Mage-D1 knockout (Mage-D1 KO)mice and littermate male wild-type (WT)mice.Micro-CT scanning was performed to observe mouse femoral bone mass,and ELISA and chemical assay were employed to detect serum levels of calcium,phosphorus,calcitonin,and parathyroid hormone in mice.After primary cultured BMSCs were identified with flow cytometry,immunofluorescence staining was utilized to detect the expression of Mage-D1 in BMSCs.BMSCs were infected by Mage-D1 silencing lentivirus,and then the cells were divided into negative control group (sh-NC)and silencing group (sh-Mage-D1).Cell scratch assay was conducted to detect the migration ability of BMSCs,and flow cytometry and CCK-8 assay were conducted to detect the cycle change and proliferation ability of BMSCs.After mineralization induction,alkaline phosphatase (ALP) staining and alizarin red staining were performed;RT-qPCR and Western blotting were used to measure the expression levels of ALP,Runx2 and Col1.RT-qPCR was used to detect mineralization-related genes p75NTR and Msx1.Results Compared with the WT mice,the femoral cortical bone thickness,cortical bone mineral content,cancellous bone mineral content,trabecular number,and cancellous bone surface density were decreased,and trabecular separation was increased in the Mage-D1 knockout homozygous mice (P<0.05).There were no significant changes in the serum levels of calcium,phosphorus,calcitonin and parathyroid hormone in mice after Mage-D1 knockout.Mage-D1 was expressed in the whole BMSCs and was highly expressed in the nucleus and perinuclear regions.Compared with the sh-NC BMSCs,the sh-Mage-D1 group had decreased proliferation ability (P<0.01),enhanced migration ability (P<0.01),and decreased expression of ALP,Runx2 and Col1 genes (P<0.05)and protein (P<0.01)after mineralization induction,milder ALP and alizarin red stain,and lower expression levels of p75NTR and Msx1.Conclusion Mage-D1 knockout can significantly reduce femur bone mass in mice.It can promote the proliferation and inhibit migration of BMSCs,and positively regulate their mineralization in vitro,and the p75NTR-Dlx1/Msx1 signaling axis may be involved in the regulation of bone metabolism by Mage-D1.


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