1.TMEM9 promotes the proliferation and invasion of prostate cancer by affecting autophagy
Huixuan ZHU ; Shuai SU ; Jindong ZHANG ; Yu LUO ; Liangdong SONG ; Hengchuan LIU ; Delin WANG
Journal of Chongqing Medical University 2025;50(9):1281-1289
Objective:To explore the expression of transmembrane protein 9(TMEM9)as an oncogene in prostate cancer(PCa),and to examine its effect on the proliferation and invasion of PCa cells as well as on the Wnt/β-catenin signaling pathway and autophagy of PCa cells by intervening with its expression.Methods:The Cancer Genome Atlas was used for pan-cancer analysis of TMEM9 expres-sion levels in different tumors,and TMEM9 protein and messenger ribonucleic acid(mRNA)levels in clinical PCa and prostatic hyper-plasia specimens were analyzed.Real-time quantitative polymerase chain reaction and western blotting were used to analyze TMEM9 expression levels in different PCa cell lines.Cell counting,terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL),and Transwell assay were used to analyze cell proliferation,apoptosis,and PCa cell invasion,respectively.Subcutaneous tu-morigenesis in nude mice was used to analyze tumor proliferation in vivo.Western blotting was used to analyze the expression of autophagy-related pathway proteins,and transmission electron microscopy and immunofluorescence colocalization were used to deter-mine the colocalization of autophagosomes and lysosomes.Results:TMEM9 was highly expressed in PCa.The mRNA and protein ex-pression levels of TMEM9 were higher in PCa tissues than in prostatic hyperplasia tissues.The expression of TMEM9 was highest in PC3 cells(human PCa cells)(t=17.150,P<0.01).In TMEM9-knocked down PC3 cells,the proliferation(t=3.165,P<0.05)and inva-sion(F=76.620,P<0.01)significantly decreased,and apoptosis(t=13.530,P=0.010)significantly increased.After knockdown of TMEM9,the volume(F=1 699.000,P<0.01)and weight(t=9.057,P<0.01)of subcutaneous tumors in nude mice were reduced,and tu-mor growth was inhibited.TMEM9 regulated the Wnt/β-catenin signaling pathway and inhibited the AKT-mTOR signaling pathway to promote autophagy in PCa cells.Conclusion:TMEM9 inhibits the AKT-mTOR signaling pathway and promotes the proliferation and invasion of PCa cells through autophagy.
2.Potential mechanisms of Ganmai Dazao Decoction for treating insomnia: an integration of network pharmacology, molecular docking, and molecular dynamics simulation
Jiaolong YU ; Xianhai LI ; Yao LIU ; Tanjun WEI ; Fei CHEN ; Delin ZHANG ; Runfeng ZHANG
Sichuan Mental Health 2025;38(6):519-527
BackgroundInsomnia, a common sleep disorder, is robustly associated with cardiovascular diseases, diabetes, and psychiatric disorders, substantially impairing quality of life. Although clinically commonly used medications are effective, long-term use may lead to drug resistance and dependence. While the efficacy of Ganmai Dazao Decoction in improving insomnia is definite, its underlying molecular mechanisms remain unclear. ObjectiveTo explore the active ingredients and core targets of Ganmai Dazao Decoction in the treatment of insomnia, systematically reveal its potential molecular pharmacological mechanism, and to provide references for clinical application. MethodsIn November 2024, the active ingredients and related targets of Ganmai Dazao Decoction were screened from the INPUT database. Insomnia-related datasets were acquired from the Gene Expression Omnibus (GEO) database, followed by differential expression analysis using GEO2R to identify differentially expressed genes (DEGs) associated with insomnia. The shared targets were obtained through Venn diagrams, and the protein-protein interaction (PPI) network was constructed using the STRING database and Cytoscape 3.9.1. Enrichment analyses were conducted on the shared targets using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). The top 3 key active ingredients and the top 10 core targets in terms of node degree values were selected. Molecular docking and molecular dynamics simulation of receptors and ligands were performed using AutoDock 4.4.6, and the results were visualized using Pymol 3.0.3 to further verify the stability of the receptor-ligand complex system. ResultsA total of 337 active ingredients and 5 265 drug-related targets in Ganmai Dazao Decoction were retrieved, along with 1 061 insomnia-related DEGs. 287 shared targets were identified between Ganmai Dazao Decoction and insomnia. The traditional Chinese medicine-active ingredients-shared targets-disease network showed that quercetin, catechins and kaempferol were the key components of Ganmai Dazao Decoction in treating insomnia. These three components alleviate insomnia by acting on ten core targets, including nuclear factor kappa B inhibitor alpha (NFKBIA), fibronectin 1 (FN1), interleukin-6 (IL6), protein c-Fos (FOS), histone acetyltransferase p300 (EP300), histone deacetylase 1 (HDAC1), transcription factor Jun (JUN), heat shock protein HSP 90-alpha 1 (HSP90AA1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and interleukin-1 beta (IL1β). GO and KEGG enrichment analyses indicated that Ganmai Dazao Decoction may alleviate insomnia through the IL17 signaling pathways, lipid and atherosclerosis signaling pathways, and other mechanisms. The results of molecular docking demonstrated strong binding affinity between the 3 key components and the 10 core targets of Ganmai Dazao Decoction. Molecular dynamics simulations further confirmed the stability of the quercetin-GAPDH, catechin-HDAC1 and kaempferol-EP300 complexes. ConclusionThe key components of Ganmai Dazao Decoction, namely quercetin, catechin, and kaempferol, exert therapeutic effects on insomnia by targeting 10 core proteins and modulating multiple pathways, including the IL17 signaling pathway, lipids and atherosclerotic-related pathways. [Funded by Chengdu Medical College Level Scientific Research Project (number, CYZYB23-01)]
3.METTL3-mediated m6A methylation regulates lipopolysaccharide-induced endothelial cell permeability changes
Jianfeng WANG ; Huilin YU ; Youxin YU ; Junhui SONG ; Chengying MENG ; Wei JIANG ; Delin HU
Acta Universitatis Medicinalis Anhui 2024;59(6):1023-1028
Objective To explore the molecular mechanism of N6-methyladenosine(m6A)methylation mediated by methyltransferase 3(METTL3)in regulating lipopolysaccharide(LPS)-induced endothelial cell permeability changes.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro.HUVECs were treated with LPS 50,125,250,500,1 000,2 000 ng/ml for 24 h.METTL3 mRNA expression was detected by Real-time PCR.After HUVECs were intervened with 500 ng/ml for 24 h,the methylation level of m6A was detec-ted,and cell permeability was measured by cell permeability test.Real-time PCR and Western blot were used to detect mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-caherin).METTL3 overexpressed stable cell lines were constructed to measure the changes of m6A methylation level and per-meability of endothelial cells during METTL3 overexpression.Results Compared to the control group,LPS inhibi-ted the expression of HUVECs METTL3 mRNA,decreased the methylation of m6A,increased the cell permeabili-ty,and decreased the mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-Caherin).When METTL3 was overexpressed,the m6A methylation levels of endothelial cells were enhanced,and the increase of endothelial cell permeability induced by LPS was reversed.Conclusion METTL3-mediated m6A methylation can improve the permeability of endothelial cells induced by sepsis.
4.Value of peripheral blood PCT, CRP, FIB and D-D levels for early diagnosing PTB complicated with bacterial pneumonia
SHEN Tian ; ZHU Huiming ; TIAN Hua ; ZHOU Yu ; ZHU Yihua ; GU Delin ; CHEN Junlin ; CAO Xingjian ; YUAN Ying
China Tropical Medicine 2023;23(7):730-
Abstract: Objective To investigate the early diagnostic value of peripheral blood procalcitonin (PCT), C-reactive protein (CRP), fibrinogen (FIB) and D-dimer (D-D) levels in patients with pulmonary tuberculosis (PTB) complicated with bacterial pneumonia. Methods A total of 102 patients who admitted to Department of Tuberculosis of Affiliated Nantong Hospital of Shanghai University from Jan 2021 to May 2022 were enrolled in this study and divided into a group (52 cases) with pulmonary tuberculosis (PTB) patients and a group (50 cases) with PTB patients complicated with bacterial pneumonia. The levels of PCT, CRP, FIB and D-D in the peripheral blood were measured, the differences and correlations in all indicators were compared among two groups. The sensitivity and specificity of these indicators in the early diagnosis of PTB complicated with bacterial pneumonia were analyzed by receiver operating characteristic (ROC) curve. Results The levels of PCT, CRP, FIB and D-D in the peripheral blood from the PTB complicated with bacterial pneumonia group were 0.06 (0.04, 0.16) ng/mL, 38.00 (3.88, 96.10) mg/L, 4.51 (3.02, 6.07) g/L, and 0.59 (0.34, 1.88) mg/L, respectively, which were significantly higher than corresponding 0.04 (0.03, 0.04) ng/mL, 3.20 (0.84, 7.22) mg/L, 2.96 (2.48, 3.77) g/L, and 0.27 (0.17, 0.36) mg/L in the PTB group (Z=-4.784, -5.233, -3.853, -4.199, all P<0.001). Furthermore, the levels of CRP and FIB in the PTB complicated by bacterial pneumonia group were highly positively correlated (r=0.855, P<0.001). The area under the ROC curve (AUC) of PCT, CRP, FIB and D-D for early diagnosis of PTB complicated with bacterial pneumonia were 0.757, 0.794, 0.747 and 0.764, respectively. In addition, the AUC obtained by simultaneous measurement of PCT, CRP, FIB and D-D was as high as 0.916, and the sensitivity and specificity of diagnosing PTB complicated with bacterial pneumonia were increased to 85.7% and 96.9%, respectively, which were higher than those of individual indicators. Conclusions Levels of peripheral blood PCT, CRP, FIB, and D-D all show varying degrees of increase in patients with PTB complicated with bacterial pneumonia, and detecting the levels of all four markers, rather than any single marker, can assist in early monitoring whether the tuberculosis patients are complicated with bacterial pneumonia.
5.Clinical efficacy of acupuncture combined with extracorporeal shock wave therapy in the treatment of piriformis syndrome
Delin CHU ; Jingxian CHEN ; Qiang ZHANG ; Chunlei WU ; Puyuan MA ; Dongsheng YU
Chinese Journal of Primary Medicine and Pharmacy 2023;30(6):901-904
Objective:To evaluate the clinical efficacy of acupuncture combined with extracorporeal shock wave therapy in the treatment of piriformis syndrome.Methods:Eighty patients with piriformis syndrome who received treatment in Huaihe Energy Occupational Disease Prevention Hospital Rehabilitation Medicine Center from February 2021 to December 2022 were included in this study. They were divided into a treatment group ( n = 40) and a control group ( n = 40) according to different treatment methods using a retrospective cohort study design. The treatment group was treated with acupuncture combined with extracorporeal shock wave therapy, once every 7 days for 3 consecutive weeks. The control group was treated with extracorporeal shock wave therapy alone once every 7 days for 3 consecutive weeks.Pre- and post-treatment visual analogue scale scores and clinical efficacy were compared between the two groups. Results:There was no significant difference in baseline data between the two groups (all P > 0.05). After treatment, the visual analogue scale score decreased in each group. After adjusting for covariates, there was no significant difference in visual analogue scale score between the two groups ( F = 112.38, P < 0.001). After adjusting the pre-treatment visual analogue scale score, the post-treatment visual analogue scale score in the treatment group was lower than that in the control group [1.417 (95% CI: 1.151-1.683)]. The total response rate in the treatment group was 97.5% (39/40), which was significantly higher than 82.5% (33/40) in the control group ( Z = -2.03, P = 0.042). Conclusion:Acupuncture combined with extracorporeal shock wave therapy is highly effective on piriformis syndrome.
6.Mechanism of TLR4/RhoA signaling pathway in endothelial cell permeability changes induced by continuous hemofiltration therapy in sepsis
Huilin Yu ; Jianfeng Wang ; Yi Liu ; Yuyao Liu ; Wei Jiang ; Chengying Meng ; Huan Wang ; Delin Hu
Acta Universitatis Medicinalis Anhui 2023;58(7):1159-1164
Objective :
To investigate the molecular mechanism of Toll⁃like receptor 4 ( TLR4)/Ras homologue A (RhoA) signaling pathway involved in regulating the effect of septic serum on vascular endothelial cell permeability
before and after continuous hemofiltration.
Methods :
The serum of 5 patients with sepsis before and after continuous hemofiltration treatment was collected , and the levels of inflammatory cytokines in serum before and after hemofiltration were detected. Human umbilical vein endothelial cells (HUVEC) were treated with serum before and after continuous hemofiltration for 24 hours. The expression of VE⁃cadherin , F ⁃actin , TLR4 and RhoA in vascular endothelial cells were detected by Western blot. A TLR4 low expression cell line was constructed to detect the effect of TLR4 low expression on the expression of VE⁃cadherin , F ⁃actin and RhoA and the permeability of endothelial cells.
Results :
After continuous blood treatment , the serum levels of TLR4 , RhoA , interleukin⁃1 (IL⁃1) , interleukin⁃6 (IL⁃6) and tumor necrosis factor⁃α (TNF⁃α ) significantly decreased. The expression levels of VE⁃cadherin , F ⁃actin , TLR4 and RhoA in the serum intervention group after continuous hemofiltration treatment significantly decreased , and the cell permeability significantly decreased. Low expression of TLR4 significantly promoted the expression of VE⁃cadherin and F ⁃actin , and inhibited the expression of RhoA protein.
Conclusion
TLR4/RhoA signaling pathway is involved in the regulation of changes in vascular endothelial cell permeability induced by septic serum after continuous hemofiltration treatment.
7.Associations of Hand-Foot-Mouth disease incidence among children of different age groups in kindergartens
DAI Yingxue, YUE Yong, YUE Limei, GUO Yu, SU Liyuan, HAN Delin
Chinese Journal of School Health 2020;41(8):1204-1207
Objective:
To explore the associations of Hand-Foot-Mouth disease (HFM) among children of different age groups in kindergartens, and to provide scientific evidence for HFM prevention and control.
Methods:
Cluster sampling was used to select 9 912 children from 40 kindergartens. The major caregivers of children were invited to participate into the study and fulfill a self-administered questionnaire that consisted of general background, hand washing and the history of HFM among children in the recent 1 year. Chi-square and binary logistic regression were applied,to analyze the influencing factors of HFM among children of different age groups.
Results:
In the 2 year-old-group, children who were male (OR=1.76, 95%CI=1.24-2.50), had the major caregiver with high school educational background or below (OR=1.54, 95%CI=1.06-2.24), had family income more than 100 000 yuan or more per year(OR=1.49, 95%CI=1.01-2.20), children whose major caregiver seldom wash hands immediately after coming home(OR=2.10, 95%CI=1.05-4.19),and 3-5 times per week(OR=2.07, 95%CI=1.26-3.41) were more likely to have HFM. In the 3 year-old-group, the more time children spent in the outdoors, the less likely they got HFM(3-5 times per week: OR=0.58, 95%CI=0.43-0.81; ≥6 times per week: OR=0.45, 95%CI=0.29-0.70). Children whose major caregiver did not use soap were more likely to have HFM(OR=1.67, 95%CI=1.11-2.49). In the 4 year-old-group, children who occasionally took toys when going out were less likely to get HFM (vs always, OR=0.57, 95%CI=0.38-0.88). No significant factors were found in the 5 year-old-group.
Conclusion
The behaviors and activities of major caregivers and children could influence the HFM incidence.To prevent HFM, it was advised that the major caregivers should wash hands immediately after coming home, and develop the habit of using soap for handwashing together with children at the earlier age, and encourage children to take more outside-door activities according to the age characteristics. Attention should be paid to avoid contaminating takeout snacks, and to clean the takeout toys timely.
8.Correlation between the distribution of intracranial and extracranial aterial lesions and risk factors in Chinese patients with ischemic stroke :a multicenter registry study
Yu TANG ; Lingyun JIA ; Yingqi XING ; Pinjing HUI ; Xuan MENG ; Delin YU ; Xiaofang PAN ; Yalan FANG ; Binbin SONG ; Chunxia WU ; Chunmei ZHANG ; Xiufang SUI ; Youhe JIN ; Jingfen ZHANG ; Jianwei LI ; Ling WANG ; Yuming MU ; Jingxin ZHONG ; Yuhong ZHU ; Heng ZHANG ; Xiaoyu CAI ; Yang HUA
Chinese Journal of Ultrasonography 2019;28(5):369-374
Objective To investigate the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in Chinese patients with ischemic stroke . Methods In this multi‐center study ,2 310 continuously inpatients with ischemic stroke diagnosed in 20 stroke screening and prevention project base hospitals from June 2015 to M ay 2016 were enrolled . Carotid ultrasonography and transcranial color‐coded sonography or transcranial Doppler were performed in all patients to confirm the presence of cerebral artery stenosis or occlusion . According to the distribution of lesions ,the subjects were divided into 2 groups :the simple intracranial artery stenosis group and the simple extracranial artery stenosis group . T he difference of risk factors between the two groups was compared . Results Of the 2 310 patients with ischemic stroke ,1 516 ( 65 .6% ) had simple intracranial artery stenosis and 794 ( 34 .4% ) had simple extracranial artery stenosis . T he incidence of anterior circulation artery stenosis was higher in the group of intracranial artery stenosis than that in the extracranial artery stenosis group ( 68 .1% vs 48 .7% , P <0 .001) . Posterior circulation artery stenosis and combined anterior with posterior circulation artery stenosis were more common in patients with extracranial artery stenosis group than those in intracranial artery stenosis group ( 36 .4% vs 22 .1% ,14 .9% vs 9 .8% ;all P <0 .001) . Univariate analysis of risk factors for stroke showed that patients with intracranial arterial stenosis had a higher prevelence of hypertension , diabetes ,obesity ,and family history of stroke ,and their systolic blood pressure ,diastolic blood pressure , body mass index ( BM I) ,fasting blood‐glucose ,glycosylated hemoglobin ,triacylglycerol ,total cholesterol , and low‐density lipoprotein cholesterol were significantly higher than those in the extracranial arterial stenosis group ( all P < 0 .05 ) . T he proportion of elderly ( ≥ 65 years old ) ,male and smokers in the extracranial arterial stenosis group was significantly higher than that in the intracranial arterial stenosis group ( all P <0 .05) . Multivariate logistic regression analysis showed that elderly ( ≥65 years old) ,male , and smoking history were independent risk factors for extracranial arterial stenosis ( OR= 2 .012 ,1 .637 , 1 .325 ,respectively ;all P <0 .05) . While hypertension ,diabetes ,less physical activity ,and high BM I levels were independent risk factors for simple intracranial arterial disease ( OR = 1 .301 ,1 .252 ,1 .248 ,1 .030 , respectively ;all P <0 .05) . Conclusions There are significant differences in the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in patients with ischemic stroke in China .
9. Regulation of hypoxia inducible factor-1α on permeability of vascular endothelial cells and the mechanism
Delin HU ; Youxin YU ; Rong LIANG ; Shunying ZHOU ; Shengliang DUAN ; Zhiyong JIANG ; Chengying MENG ; Wei JIANG ; Huan WANG ; Yexiang SUN ; Linsen FANG
Chinese Journal of Burns 2019;35(3):209-217
Objective:
To investigate the regulation of hypoxia-inducible factor-1α (HIF-1α) on permeability of rat vascular endothelial cells and the mechanism.
Methods:
Twelve male Sprague-Dawley rats aged 35 to 38 days were collected and vascular endothelial cells were separated and cultured. The morphology of cells was observed after 4 days of culture, and the following experiments were performed on the 2nd or 3rd passage of cells. (1) Rat vascular endothelial cells were collected and divided into blank control group, negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group according to the random number table (the same grouping method below), with 3 wells in each group. Cells in negative control group, HIF-1α interference sequence 1 group, HIF-1α interference sequence 2 group, and HIF-1α interference sequence 3 group were transfected with GV248 empty plasmid, recombinant plasmid respectively containing HIF-1α interference sequence 1, interference sequence 2, and interference sequence 3 with liposome 2000. Cells in blank control group were only transfected with liposome 2000. After transfection of 24 h, expression levels of HIF-1α mRNA and protein of cells in each group were respectively detected by reverse transcription real-time fluorescent quantitative polymerase chain reaction and Western blotting (the same detecting methods below) . The sequence with the highest interference efficiency was selected. (2) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α low expression group, with 3 wells in each group. Cells in blank control group were only transfected with liposome 2000, and cells in negative control group and HIF-1α low expression group were respectively transfected with GV248 empty plasmid and low expression HIF-1α recombinant plasmid selected in experiment (1) with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α in each group were detected. (3) Another batch of rat vascular endothelial cells were collected and divided into blank control group, negative control group, and HIF-1α high expression group, with 3 wells in each group. Cells in blank control group were transfected with liposome 2000, and cells in negative control group and HIF-1α high expression group were respectively transfected with GV230 empty plasmid and HIF-1α high expression recombinant plasmid with liposome 2000. After 14 days of culture, the mRNA and protein expressions of HIF-1α of cells in each group were detected. (4) After transfection of 24 h, cells of three groups in experiment (1) and three groups in experiment (2) were collected, and mRNA and protein expressions of myosin light chain kinase (MLCK), phosphorylated myosin light chain (p-MLC), and zonula occludens 1 (ZO-1) of cells were detected. Data were processed with one-way analysis of variance and
10. Influence of three-level collaboration network of pediatric burns treatment in Anhui province on treatment effects of burn children
Zhengguo XIA ; Xianliang ZHOU ; Weichang KONG ; Xingzhao LI ; Junhui SONG ; Linsen FANG ; Delin HU ; Chen CAI ; Yizhong TANG ; Youxin YU ; Chunhua WANG ; Qinglian XU
Chinese Journal of Burns 2018;34(3):143-148
Objective:
To explore the influence of three-level collaboration network of pediatric burns in Anhui province on treatment effects of burn children.
Methods:
The data of medical records of pediatric burn children transferred from Lu′an People′s Hospital and Fuyang People′s Hospital to the First Affiliated Hospital of Anhui Medical University from January 2014 to December 2015 and January 2016 to September 2017 (before and after establishing three-level collaboration network of pediatric burns treatment) were analyzed: percentage of transferred burn children to hospitalized burn children in corresponding period, gender, age, burn degree, treatment method, treatment result, occurrence and treatment result of shock, and operative and non-operative treatment time and cost. Rehabilitation result of burn children transferred back to local hospitals in 2016 and 2017. Data were processed with


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