1.Bioinformatics Analysis and Validation of Cuproptosis-related Genes in Wilson Disease
Zhuang TAO ; Meixia WANG ; Shuai KANG ; Jipeng LIU ; Rui WANG ; Jiafeng ZHOU ; Wenming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):124-131
ObjectiveTo explore the role of cuproptosis and identify cuproptosis-related genes in Wilson disease (WD) through bioinformatics analysis and clinical validation,providing implications and directions for the diagnosis and treatment of WD. Methods(1) Screening of target genes: The differentially expressed genes (DEGs) between WD and healthy control were obtained from GeneCards,and the cuproptosis-related genes were obtained from Gene Expression Omnibus (GEO) and published literature.The cuproptosis-related genes in WD were obtained by intersection.Through gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses,the specific biological process,functions or metabolic pathways of cuproptosis-related genes in WD were predicted.Molecular docking and PyMOL visualization were then performed to analyze and verify the potential regulatory mechanism of Gandou Fumu Decoction for cuproptosis.(2)Validation of target genes: The blood samples of 15 WD patients treated in the department of encephalopathy and 15 healthy volunteers undergoing physical examinations in the health management center were randomly collected from the First Affiliated Hospital of Anhui University of Chinese Medicine.The expression levels of target genes were determined by Western blot and real-time PCR. Results(1) A total of 3 607 DEGs in WD were obtained from GSE107323 in GEO,and 68 cuproptosis-related genes were obtained from GeneCards and published literature.Twelve common target genes were obtained by intersection,including three up-related genes(SQSTM1,MIF1,and TAX1BP1) and nine down-regulated genes(CP,SERPINE1,AOC3,GPX4,SLC27A5,VEGF-A,PDHB,PDK1,and ATP7B).The common target genes were mainly enriched in monocarboxylic acid metabolism,oxidoreductase activity,negative regulation of molecular functions,which mainly involved HIF-1,ferroptosis and other signaling pathways.Molecular docking and PyMOL visualization results showed Gandou Fumu Decoction had good binding ability with the cuproptosis-related genes PDK1,SERPINE1,VEGFA,and AOC3 in WD.(2)A total of 30 blood samples were collected,including 15 WD patients and 15 health volunteers.Western blot results showed that expression levels of target genes were consistent with the results obtained by bioinformatics analysis.RT-qPCR results showed that compared with healthy volunteers,WD patients had down-regulated mRNA levels of SERPINE1,GPX4,SLC27A5,and VEGF-A and up-regulated mRNA levels of SQSTM1 and MIF1(P<0.05). ConclusionThe expression levels of cuproptosis-related genes in WD patients are consistent with the results predicted by bioinformatics analysis.The characteristic preparation Gandou Fumu Decoction of Xin'an Medicine showed good binding abilities with the cuproptosis-related genes in WD.Cuproptosis may play a key role in the pathophysiological mechanism of WD,which can provide a new target for the diagnosis and treatment of WD.
2. Effective dose of esketamine for prevention on propofol injection pain in painless abortion
Yanping SHEN ; Lijun YIN ; Haiya YAN ; Wenming ZHUANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2022;27(6):660-664
AIM: To investigate the effective dose of esketamine for prevention on propofol injection pain in painless abortion. METHODS: From November 2021 to December 2021, thirty patients undergoing painless abortion, aged 20 to 40 years old, ASA physical status or Ⅱ, BMI 19-26 kg/m
3.Construction and identification of interference plasmid targeting on TNFAIP8
Wenming LIU ; Jingjing YANG ; Ruyi HU ; Xingfeng QIU ; Chunyan SHI ; Zhongquan QI ; Zhongchen LIU ; Guohong ZHUANG
Chinese Journal of Immunology 2015;(5):650-654
Objective:To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-pSIREN-RetroQ.Methods:Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-pSIREN-RetroQ interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results:We successfully design and built three TNFAIP8-shRNA-pSIREN-RetroQ interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.

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