目的 探讨补肾活血法预防芳香化酶抑制剂(aromatase inhibitors,AIs)所致骨丢失的关键靶标和通路.方法 将小鼠随机分为假手术组、模型组和给药组,模型组和给药组去势后予以 10 μg/d来曲唑溶液皮下注射构建绝经后AIs所致骨质疏松动物模型,给药组同时予以黄芪补肾活血汤(HQD)19.24 g/(kg·d)灌胃预防骨丢失,假手术组与模型组予以等量生理盐水灌胃.给药三个月后取小鼠股骨进行骨密度检测验证造模效果,并借助TMT蛋白组学测序分析获取其组学特征.结果 与假手术组相比,模型组小鼠骨小梁结构疏松紊乱,数量减少,证明造模成功.与模型组相比,给药组骨小梁粗大致密,数量增多,证明补肾活血法预防作用显著.模型组与假手术组相比,上调蛋白 275 个,下调 189 个;给药组与模型组相比,上调蛋白286 个,下调蛋白 898 个.三组对比后共有 18 个交集蛋白,分别为 Ptbp1、Epx、Ear1、Ear2、Col6a5、Mta2、Alox15、Lztfl1、Ccl6、Diaph2、Wfdc21、Smarcb1、Strip1、C19orf12、Pex14、Akr1b7、Fahd1、Nr1d2.差异蛋白主要参与了细胞发育和分化的调节、脂质代谢、核酸内切酶活性等生物过程,并在铁死亡、花生四烯酸代谢通路、肌钙蛋白细胞骨架调节、PI3K-AKT等信号通路显著富集.结论 补肾活血法可有效预防AIs所致骨丢失的发生,其关键靶标和作用机制可能与Ptbp1、Alox15 及铁死亡、PI3K-AKT等通路相关.

目的:亮氨酸拉链转录因子1(LZTFL1)是一种与纤毛信号转导相关的蛋白质,关于其如何在信号转导中发挥作用以及作用机制目前尚不清楚.莱茵衣藻(Chlamydomonas reinhardtii)是研究纤毛信号传导的模式生物,而目前对莱茵衣藻LZTFL1蛋白的研究甚少,尚未有相应的检测抗体,因此制备LZTFL1多克隆抗体用于后续实验研究.方法:采用RT-PCR技术从C.reinhardtii CC125中提取总RNA,扩增981bp的目的基因Lztfl1,插入到pET-28a(+)原核表达载体,成功构建pET-28a(+)-Lztfl1重组质粒,转入E.coli BL21(DE3)经IPTG诱导后成功表达6×His-LZTFL1融合蛋白,融合蛋白经亲和纯化后免疫新西兰大白兔制备多克隆抗体,最后采用间接ELISA法测得抗血清效价达到1∶512 000,经Westem blot对C.reinhardtii CC125检测具有较高特异性.结果:首次实现了莱茵衣藻LZTFL1蛋白的原核表达,制备出一支兔抗莱茵衣藻LZTFL1蛋白的多克隆抗体,为后续研究LZTFL1蛋白在莱茵衣藻中的结构功能及在纤毛信号转导中的相互作用奠定了基础.

Leptin receptor (LepRb) signaling pathway in the hypothalamus of the forebrain controls food intake and energy expenditure in response to an altered energy state.Defects in the LepRb signaling pathway can result in leptin-resistance and obesity.Leucine zipper transcription factor like 1 (Lztfl1)/BBS17 is a member of the Bardet-Biedl syndrome (BBS) gene family.Human BBS patients have a wide range of pathologies including obesity.The cellular and molecular mechanisms underlying Lztfl1-regulated obesity are unknown.Here,we generated Lztfl1f/f mouse model in which Lztfl1 can be deleted globally and in tissue-specific manner.Global Lztfl1 deficiency resulted in pleiotropic phenotypes including obesity.Lztfl1-/-mice are hyperphagic and showed similar energy expenditure as WT littermates.The obese phenotype of Lztfl1-/-mice is caused by the loss of Lztfll in the brain but not in the adipocytes.Lztfl1-/-mice are leptin-resistant.Inactivation of Lztfl1 abolished phosphorylation of Stat3 in the LepRb signaling pathway in the hypothalamus upon leptin stimulation.Deletion of Lztfl1 had no effect on LepRb membrane localization.Furthermore,we observed that Lztfl1-/-mouse embryonic fibroblasts (MEFs) have significantly longer cilia than WT MEFs.We identified several proteins that potentially interact with Lztfl1.As these proteins are known to be involved in regulation of actin/cytoskeleton dynamics,we suggest that Lztfl1 may regulate leptin signaling and ciliary structure via these proteins.Our study identified Lztfl1 as a novel player in the LepRb signaling pathway in the hypothalamus that controls energy homeostasis.

目的 探讨miR-208a-3p靶向亮氨酸拉链转录因子样1(LZTFL1)对宫颈癌细胞增殖、侵袭转移的影响和分子机制.方法 收集2016年5月～2018年3月于湖北省中西医结合医院妇产科收治并经病理科确诊的40例宫颈癌患者手术标本.实时荧光定量PCR(RT-qPCR)和蛋白质印记(Western Blot)检测宫颈癌组织和与其对应的癌旁组织中miR-208a-3p和LZTFL1的表达.将将宫颈癌细胞Hela分为anti-miR-NC组(转染anti-miR-NC)、anti-miR-208a-3p组(转染anti-miR-208a-3p)、pcDNA组(转染pcDNA)、pcDNA-LZTFL1组(转染pcDNA-LZTFL1).采用MTT法检测细胞活力,Transwell实验检测细胞迁移和侵袭数目,western blot检测细胞周期素D1 (CyclinD1)、基质金属蛋白酶2(MMP-2)和MMP-9蛋白的表达.结果 相较于癌旁组织,宫颈癌组织中miR-208a-3p的表达显著升高,LZTFL1的表达显著降低(P＜0.05).miR-208a-3p靶向LZTFL1并负性调控其表达.与anti-miR-NC组比较,anti-miR-208a-3p组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白的表达显著降低,p21的表达显著升高,差异均有统计学意义(P＜0.05).与pcDNA组比较,pcDNA-LZTFL1组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白表达显著降低,p21表达显著升高,差异均有统计学意义(P＜0.05).与anti-miR-208a-3p+ si-NC组比较,anti-miR-208a-3p+ si-LZTFL1组Hela细胞活力、迁移侵袭能力、CyclinD1、MMP-2和MMP-9蛋白的表达显著升高,p21的表达显著降低,差异均有统计学意义(P＜0.05).结论 宫颈癌中miR-208a-3p呈高表达,LZTFL1呈低表达.抑制miR-208a-3p通过上调LZTFL1可抑制宫颈癌细胞的增殖、迁移和侵袭.

目的 探讨刺槐素对肺癌细胞增殖、迁移和侵袭的影响及分子机制.方法 用浓度分别为4、8、16 μmol/L的刺槐素处理A549细胞,作为低、中、高浓度组,不作任何处理的细胞作为对照组;将pcDNA、pcDNA-LZTFL1转染至A549细胞中,记为pcDNA组、pcDNA-LZTFL1组;将si-NC、si-LZTFL1转染至A549细胞中再用16 μmol/L的刺槐素处理,记为刺槐素+ si-NC组、刺槐素+si-LZTFL1组.四甲基偶氮唑盐比色法(MTT)检测细胞增殖抑制率;Western blotting法检测LZTFL1、CyclinD1、p21、MMP-2、MMP-9蛋白表达;Transwell检测细胞迁移和侵袭;实时荧光定量PCR (qRT-PCR)检测LZTFL1mRNA表达水平.结果 与对照组比较,刺槐素低、中、高浓度处理的肺癌A549细胞中细胞增殖抑制率显著升高,细胞迁移、侵袭数显著降低,CyclinD1、MMP-2、MMP-9蛋白表达水平显著降低,p21蛋白表达水平显著升高,LZTFL1 mRNA和蛋白表达水平显著升高(P＜0.05).过表达LZTFL1抑制肺癌细胞的增殖、迁移和侵袭.干扰LZTFL1表达逆转了刺槐素对肺癌A549细胞增殖、迁移和侵袭的抑制作用.结论 刺槐素可能通过上调LZTFL1的表达抑制肺癌细胞的增殖、迁移和侵袭.

Dear Editor,Clinical outcomes of SARS-CoV-2 infection are highly heterogeneous,ranging from symptoms-free infection to death of the patient (Park et al.2020).Although the out-come can be explained to some extent by age and comor-bidities,genetic factors have also been linked to the prognosis of SARS-CoV-2 infection (Hu et al.2021;Liu et al.2020;Maya et al.2020;Pairo-Castineira et al.2021;The Severe COVID-19 GWAS Group 2020;Zeberg and P(a)(a)bo 2020).To this date,the association of nine single nucleotide polymorphisms (SNPs) at genome-wide signif-icance level has been reported in the Genome Wide Association Study (GWAS) catalog for severe COVID-19 outcome phenotype (MONDO_0100096) (GWAS catalog2021).