1.cDNA Cloning and Sequence Analysis of Major Allergen of Dermatophagoides farinae (Der f 2) in South China
Minqi HAO ; Jun XU ; Nanshan ZHONG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To clone and analyze the cDNA of major allergen Der f 2 of Dermatophagoides farinae in south China. Methods cDNA of Der f 2 was cloned by RT-PCR, screened and their sequences were analyzed. Results cDNAof Der f 2 was cloned. The sequence of the cloned Der f 2 was different with that published (D10448) in GenBank, with 87 additional nucleotides inserted into the 62th nucleotide of the original one. According to the original ORF, the deduced amino acids that were located prior or after the inserted 29 amino acid sequence showed no changes. Conclusion The cDNA of Der f 2 was cloned from Dermatophagoides farinae and its sequence showed significant difference with that reported in the GenBank.
2.Preventive effect of Gehua Jiecheng Decoction in IL-6/JAK2/STAT3 pathway in"inflammation-cancer transformation"of ulcerative colitis with spleen deficiency and damp-heat
Xiaoling LI ; Yuhong WU ; Hailong LI ; Yinxia YIN ; Yali SHE ; Minqi HAO ; Yonglin LIANG
Chinese Journal of Immunology 2024;40(7):1454-1460,1466
Objective:To investigate effect of Gehua Jiecheng Decoction on IL-6/JAK2/STAT3 signaling pathway in colon tissue of ulcerative colitis with"inflammatory to cancer transformation"(UC-UCAC)mice of spleen deficiency and damp-heat type.Methods:Ten from 80 SPF C57BL/6 male mice were randomly selected as blank group and other 70 mice were selected as model group.After establishment of spleen deficient damp-heat model,mice were randomly divided into model group(1st,2nd and 3rd cycle),Gehua Jiecheng Decoction high,medium and low doses groups and mesalazine group,with 10 pieces per group.UC-UCAC transformation model was further established with azo methane oxide solution(AOM)/sodium glucan sulfate(DSS).Each group was treated with corresponding drugs for 4 weeks.General state of mice was observed.Score of disease activity index(DAI)was calculated.HE staining was used to observe colonic mucosa pathology of mice.EGFR,IL-6,JAK2,STAT3 and p-STAT3 proteins and genes expressions in mice colon tissues were detected by Western blot,IHC and RT-qPCR,respectively.Results:Compared with blank group,mice in model group(3rd cycle)were generally in a worse state,colon mucosal tissue was cancerous,DAI score,target proteins and genes expressions were significantly increased(P<0.01).Compared with model group(3rd cycle),general state of mice in all treatment groups were restored and colonic tissues pathology were improved to some extent.Target proteins and genes expressions in other treatment groups were significantly decreased except for Gehua Jiecheng Decoction low-dose group(P<0.01).Conclusion:Gehua Jiecheng Decoction may destroy tumor inflammatory microenvironment,repair damaged colonic mucosa tissue,delay inflamma-tion-cancer transformation process and prevent UCAC by inhibiting activation of IL-6/JAK2/STAT3 signaling pathway.
3.Study on the Mechanism of Astragali Radix in the Treatment of Ulcerative Colitis Based on Network Pharma- cology and Molecular Docking
Minqi HAO ; Jiahui WANG ; Xiaoling LI ; Hailong LI ; Yuhong WU
China Pharmacy 2021;32(10):1215-1223
OBJECTIVE:To predict the potenti al target and mechanism of Astragali Radix in the treatment of ulcerative colitis (UC),and to provide reference for the clinical application of Astragali Radix in the treatment of UC. METHODS :The active components and their corresponding target genes of Astragali Radix were retrieved by TCMSP and UniProt KB database.related target genes of UC were searched by Gene Cards GZK-2018-5) database. The intersection target genes of Astragali Radix and were obtained by Venny 2.1.0 online mapping tool ,and interaction network of “drug-compound-intersection target ” was constructed by using Cytoscape 3.7.0 software. PPI network of intersecting targets was obtained by using STRING 结合动物模型。E-mail:172924249@qq.com database, and the visualization analysis and topological analysis w ere carried out by using Cytoscape 3.7.0 software to obtain the core target genes. By using DAVID database ,the gene ontology (GO) function annotation and KEGG pathway enrichment of intersecting target genes were carried out ,and the “target-pathway”enrichment network was constructed by using Cytoscape 3.7.0 software. Through Auto Dock vina 1.1.2 software, the top five active components in the list of degree value were linked with the protein encoded by the core target genes ;Discovery Studio 3.5 software was applied to draw out binding pattern map. RESULTS :There were 143 compounds in Astragali Radix ,20 active components were screened out ,and 189 corresponding target genes were selected ;there were 4 356 UC disease related target genes. There were 126 intersection target genes of Astragali Radix (involving 14 active components )and UC. The core target genes in PPI network were AKT1,MAPK1,RB1,JUN,etc. A total of 2 294 GO items (q value<0.05)were obtained from GO functional annotation ,including 2 093 biological process items (e.g. response to lipopolysaccharide ,response to molecule of bacterial origin ),49 cell composition items (e.g. membrane raft ,membrane microdomain ),and 152 molecular function items (e.g. nuclear receptor activity ,ligand-activated transcription factor activity ). KEGG pathway enrichment analysis yielded 160 items(q value<0.05),such as fluid shear stress and atherosclerosis signaling pathway ,phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signaling pathway ,interleukin-17 (IL-17) signaling pathway. Molecular docking results showed that top 5 active ingredients (quercetin,kaempferol,formenonetin,isorhamnetin,7-O-methylisomucronulatol) in the list of degree value had binding energies <5.0 kcal/mol with the protein encoded core targets. CONCLUSIONS :Quercetin,kaempferol,formononetin and other active components in Astragali Radix may play a role in the treatment of UC through the action of MAPK14,JUN,AKT1 and other target genes ,and then on the signal pathways such as PI 3K/Akt and IL- 17.