经HIV包膜蛋白gp120刺激的T细胞外泌体促进巨噬细胞M2极化

目的:本研究旨在探讨经HIV膜蛋白gp120处理后人T淋巴细胞系(H9)外泌体对巨噬细胞极化的影响。方法:采用CCK8试剂盒检测gp120处理后人T淋巴细胞H9活力;采用ELISA法检测H9细胞上清液中炎症因子水平;通过电镜和Western blot实验鉴定外泌体特征;PHK67染色观察外泌体进入巨噬细胞情况;最后通过ELISA和免疫荧光检测巨噬细胞极化标记物,分析gp120处理后T细胞外泌体对巨噬细胞极化的影响。结果:HIV gp120蛋白抑制人T淋巴细胞H9增殖并促进炎症因子释放。成功提取人T淋巴细胞H9外泌体,电镜下为中间凹陷的膜结构,高表达标记蛋白CD9、CD63、CD81。PHK67染色结果显示H9细胞外泌体可进入巨噬细胞。经gp120处理的H9细胞外泌体可促进巨噬细胞向M2型极化。结论:HIV膜蛋白gp120处理人T淋巴细胞H9分泌的外泌体能够促进巨噬细胞M2极化,可能是gp120在免疫调节中的新机制。

Extracellular vesicles from T cells stimulated by HIV envelope protein gp120 promote macrophage M2 polarizatio

Objective:To investigate the effects of exosomes from human T lymphocyte line(H9)treated with HIV envelope protein gp120 on macrophage polarization.Methods:The viability of gp120-treated H9 T lymphocytes was as-sessed using the CCK8 assay.Inflammatory cytokine levels in the supernatant of H9 cells were determined by ELISA.Exosome characteristics were identified through electron microscopy and Western blot experiments.PHK67 staining was employed to observe the uptake of exosomes by macrophages.Finally,macrophage polarization markers were detected using ELISA and immuno-fluorescence to analyze the impact of gp120-treated T cell exosomes on macrophage polarization.Results:HIV gp120 protein inhibited the proliferation of human T lymphocytes H9 and promoted the release of inflammatory cytokines.Exosomes from H9 T lymphocytes were successfully isolated,displaying a cup-shaped membranous structure under electron microscopy and overexpressing marker proteins CD9,CD63,and CD81.PHK67 staining results indicated that exosomes from H9 cells could be internalized by macrophages.Exosomes from gp120-treated H9 cells promoted the polarization of macrophages towards the M2 pheno-type.Conclusion:Exosomes secreted by human T lymphocytes H9 treated with HIV envelope protein gp120 can promote M2 polarization of macrophages,suggesting a potential novel mechanism of gp120 in immune modulation.

mir-3168 targeted inhibition of TP53 promotes malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells

Objective:To investigate the effect of mir-3168 on the malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells,and to verify its target gene.Methods:The expression of mir-3168 in AGS and AGS/DDP gastric cancer cells was detected by qPCR,and mir-3168 mimic,inhibitor and negative control were synthesized.They were transfected into AGS and AGS/DDP gastric cancer cells,respectively.The expression of mir-3168 and TP53 mRNA was detected by qPCR.Cell viability was detected by CCK8 under gradient cisplatin treatment and non treatment,apoptosis was detected by flow cytometry,cell invasion was detected by Transwell,and TP53 protein expression was detected by western blot,The database predicted the binding sites of mir-3168 and TP53.According to the binding sites,the double luciferase experiment was used to verify the binding of mir-3168 and TP53.Results:Compared with cisplatin sensitive gastric cancer cell AGS,mir-3168 was significantly overexpressed in cisplatin resistant gastric cancer cell AGS/DDP;mir-3168 mimic promotes cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and inhibits apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 inhibitor inhibits cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and promotes apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 mimic inhibits the expression of TP53 mRNA and protein,and mir-3168 inhibitor promotes the expression of TP53 mRNA and protein;Targetscan database predicted that there was a binding point between mir-3168 and TP53,and the double luciferase experiment suggested that mir-3168 was bound to TP53 through the predicted binding site.Conclusion:mir-3168 may promote the malignant transformation of AGS and AGS/DDP gastric cancer cells and cisplatin resistance by targeting TP53.

桂西地区869例慢性乙型肝炎病毒基因分型与耐药突变分析

目的乙型肝炎病毒(HBV)基因型及耐药突变情况与疾病进展和药物疗效密切相关,有效开展其测定具有重要意义。文中旨在检测桂西地区869例HBV的基因型及耐药突变基因,为临床抗HBV治疗提供依据。方法收集2016年1月至2018年3月右江民族医学院附属医院感染性疾病科门诊及住院的来自桂西地区且HBV DNA>1.0×103IU/mL的869例慢性乙型肝炎病毒携带(CHB)患者。采用PCR和反向点杂交法技术检测其HBV基因分型与耐药突变情况。结果共检出7种基因分型,分别为B基因型304例(34.98%),C基因型268例(30.84%),D基因型147例(16.92%)及B+C基因型、B+D基因型、C+D基因型、B+C+D基因型。63例患者发生耐药突变,耐药突变率为7.25%,其中C基因型31例(49.21%),B基因型19例(30.16%);突变模式有rt204I、rt181V、rt236T、rt180M+rt204V+rt204I等14种,其中rt180M+rt204V突变频率最高(17.46%)。B、C基因型的总体突变率(19%、31%)差异有统计学意义(P<0.05)。结论桂西地区CHB患者HBV基因型多样,主要以B、C基因型为主,且耐药突变率较高,突变模式复杂,这一结果为桂西地区临床抗HBV治疗提供了依据。

基于lncRNA-mRNA共表达网络的HBV相关肝癌生物靶点筛选及综合分析

目的 关于lncRNAs作为预测性生物标志物和治疗靶点的研究仍然非常有限。文中旨在筛选出HBV相关肝癌差异表达lncRNA谱,寻找灵敏性高、特异性好的HBV相关肝癌早期诊断分子标志物。方法 从GEO下载HBV相关肝癌芯片数据集GSE55092、GSE19665和GSE84402,筛选出差异表达的lncRNA和mRNA,并对差异基因进行功能分析。收集2019年2月至2020年12月右江民族医学院附属医院肝胆外科及体检科患者。肿瘤组(n=45)为HBV阳性原发性肝癌患者,对照组(n=38)为乙肝病毒携带无肿瘤患者。筛选差异表达基因,及HBV相关HCC的候选诊断lncRNA生物标志物,利用DAVID对共表达的差异基因进行GO富集分析及KEGG分析,利用qRT-PCR检测候选lncRNA在血浆样本的相对表达量,通过ROC曲线评价单个血浆lncRNA及与甲胎蛋白联合在HBV相关肝癌诊断中的价值。结果 随机森林的特征变量筛选出上调的AC093642.1、AL445524.1、TRIM52-AS1、EHMT2-AS1、AL356056.2,及下调的AC003991.1、LINC00844、LINC01018、AC008040.1作为HBV相关肝癌的候选诊断性lncRNA生物标志物。候选lncRNA与126个mRNA存在共表达,共199个mRNA-lncRNA共表达对。共表达mRNA的GO功能富集结果表明,表达失调的基因主要富集于与单羧酸代谢过程、类固醇代谢过程、细胞分裂、有丝分裂细胞周期过程等。KEGG通路分析显示,差异基因主要富集在p53信号通路、视黄醇代谢、PI3K-Akt信号通路、化学致癌作用和过氧化物酶体等信号通路。AC003991.1、AL445524.1、LINC00844、AL56056.2、AC008040.1、TRIM52-AS1、LINC01018肿瘤组与对照组比较差异有统计学意义,肿瘤组表达上调(P<0.05)。ROC曲线分析结果显示,AC003991.1、AL445524.1、LINC00844和LINC01018 4种lncRNA联合构建logistic回归模型的AUC为0.910,敏感度和特异度分别为0.828、0.911;4-lncRNA与甲胎蛋白联合构建logistic回归模型的AUC为0.986,敏感度和特异度分别为0.969、0.964。结论 血浆lncRNA AL356056.2、AL445524.1、TRIM52-AS1、AC008040.1、AC003991.1、LINC00844和LINC01018可作为HBV相关肝癌临床诊断潜在的循环标志物;AL445524.1、AC003991.1、LINC00844、LINC01018联合及与甲胎蛋白联合,对HBV相关肝癌具有早期诊断价值。