沉默FOXC2对TGF-β1诱导的MCF-7细胞上皮-间质转化的逆转作用

 目的： 探讨沉默叉头框C2(FOXC2)表达对转化生长因子β1（TGF-β1）诱导的人乳腺癌MCF-7细胞上皮-间质转化(EMT)的逆转作用和对乳腺癌侵袭性的影响。方法： 将体外培养的MCF-7细胞用5 μg/L TGF-β1处理,相差显微镜观察细胞形态学变化;免疫荧光检测EMT相关标志物表达。慢病毒介导的FOXC2-siRNA载体转染至TGF-β1诱导成功的MCF-7细胞,以RT- PCR和Western blotting方法检测FOXC2及EMT相关标志物上皮性钙黏蛋白(E-cad)、紧密连接蛋白 1(CLDN1)、纤维连接蛋白 1(FN1) mRNA及蛋白表达;Transwell小室模型检测沉默FOXC2表达对TGF-β1诱导的MCF-7细胞侵袭性的影响。结果： TGF-β1可诱导MCF-7细胞向间质样细胞表型转换,并上调间质样标志物FN1表达和下调上皮样标志物E-cad及CLDN1表达;而沉默FOXC2表达可逆转已间质化的MCF-7细胞恢复至上皮表型,并降低其侵袭性。结论： TGF-β1可诱导人乳腺癌MCF-7细胞发生EMT并增加其侵袭性,且TGF-β1这种诱导作用能被FOXC2-siRNA所逆转,并降低细胞的侵袭能力。     

CTNND2基因敲除对小鼠小脑发育及运动功能的影响

目的 探讨CTNND2基因敲除对小鼠小脑神经元发育和运动功能的影响及可能的机制。 方法 小鼠分为2组,每组10只,均为7周龄：野生型 (WT) C57BL/6J 小鼠为对照组,CTNND2基因敲除纯合子 (CTNND2-/-) 小鼠为实验组,PCR检测CTNND2-/- 小鼠的基因型。平衡木实验、悬挂实验和步态分析实验检测两组运动功能;免疫荧光染色、高尔基染色检测小脑普肯耶细胞的变化;Western blotting检测突触相关蛋白磷酸化突触蛋白1（p-Syn1）、突触蛋白1（Syn1）、ELKS和突触后致密蛋白 95（PSD95）以及磷脂酰肌醇-3激酶（PI3K）、磷酸化蛋白激酶B（p-Akt）、蛋白激酶B（Akt）、磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）和哺乳动物雷帕霉素靶蛋白（mTOR）的表达水平。 结果 与WT组相比,CTNND2-/-小鼠通过平衡木的时间延长,通过平衡木的比例下降,悬挂时间缩短,悬挂得分减少,前爪步幅长度和后爪步幅长度均缩短;CTNND2-/-小鼠小脑中普肯耶细胞数及其树突的分支数均减少;p-Syn1/Syn1、p-Akt/Akt和p-mTOR/mTOR比值以及ELKS、PSD95和PI3K表达水平均低于WT组。 结论 CTNND2基因敲除可能通过下调PI3K/Akt/mTOR 信号通路,影响小脑普肯耶细胞的数量和树突结构以及突触相关蛋白的合成,导致小脑发育障碍,从而影响小鼠的运动功能。     

唐氏综合征胎儿全基因组miRNA表达谱特征及其染色体分布

目的研究唐氏综合征(DS)胎儿全基因组miRNA表达谱及其编码基因的染色体分布特征。方法采用Illumina深度测序技术对6例DS胎儿(DS组)和6例正常胎儿脐带血(对照组)单个核细胞小RNA测序比对分析,确定DS全基因组miRNA表达谱及其编码基因的染色体分布特征。结果两组共检测到395种miRNA,编码于316个miRNA基因。其中149种miRNA在两组中的表达具有显著性差异(DS组中6种上调,143种下调),有51种miRNA特异性表达于对照组。21号染色体编码的14个miRNA基因在DS组中有1个(miR-802)高表达,4个(let-7c、miR-99a、miR-125b和miR-155)低表达,余下9个在两组样本中均未表达。两组样本miRNA基因表达的染色体分布趋于一致,但miRNA基因表达丰度的分布却不尽相同:DS组主要分布于8、16、17和21号染色体,对照组主要分布于3、8、14、16、17和22号染色体。结论 DS胎儿脐带血单个核细胞具有其特定的miRNA表达谱和染色体分布特征,表达丰度差异分布的染色体编码miRNA可能在DS各临床性状的形成过程中具有重要作用。     

279例药物性肝损伤自发报告分析

目的：了解药物性肝损伤在自发报告中分布情况及相关药品,为临床提供风险点参考。方法：基于2016-2018年解放军ADR数据库中70 709例自发报告数据,回顾性分析目标患者的性别、年龄、致病药物及预后情况。结果：2838例肝胆损害报告中,筛选出RUCAM量表≥6分的药物性肝损伤279例（9.83%）,男172例（61.65%）,女107例（38.35%）。排名前5位的药物类别是抗感染药物（39.78%）、心血管系统用药（16.85%）、抗肿瘤药物（15.05%）、代谢及内分泌系统用药（5.73%）、中药（5.73%）;排名前5的品种是阿托伐他汀、莫西沙星、利福平、左氧氟沙星、头孢哌酮舒巴坦。平均潜伏期27.52 d,以40~59岁患者的平均潜伏期最长（51.10 d）。结论：严重的药物性肝损伤占比相对低,呈潜伏期长、男性多于女性、涉及药物分布广等特点;提示长期使用具潜在肝毒性药物时应监测肝功能;信息详实的ADR报告数据可以满足RUCAM量表的评价标准。     

Effect of activating transcription factor 6 on the apoptosis of podocytes stimulated with palmitic acid

Objective To observe the effect of ATF6 on the apoptosis and proliferation of podocytes induced by palmitic acid (PA). Methods Podocytes were stimulated with different doses of PA for 24 h. The expression of cleaved-caspase3 was detected by Western blotting. The podocyte apoptosis was analyzed by flow cytometry (FCM), and the expression of ATF6 was tested by Western blotting and immunofluorescence staining. After the transfection of adenovirus siRNA against ATF6, the proliferation, the cell cycle and apoptosis of potocytes stimulated with PA were tested by MTT or FCM. Results The levels of cleaved-caspase3 and ATF6 of podocytes stimulated with PA were significantly increased by a dose-dependent manner compared with the control group (P＜0.05). The apoptosis of podocytes stimulated with PA was increased (P＜0.05). Compared with the podocytes stimulated with PA, the apoptosis of podocytes transfected by adenovirus siRNA against ATF6 with PA stimulation was significantly reduced (P＜0.05). The proliferation of podocytes transfected by adenovirus siRNA against ATF6 and stimulated with PA, however, was obviously increased compared with the podocytes stimulated with PA (P＜0.05). Conclusion ATF6 mediated the apoptosis of podocytes induced by palmitate acid.     

CD36 palmitoylation disrupts free fatty acid metabolism and promotes tissue inflammation in non-alcoholic steatohepatitis

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睾酮对青春期大鼠病理性攻击行为及前额叶皮质PSD-95和GAP-43表达的影响

目的 探讨去势后睾酮水平对青春期大鼠病理性攻击行为和前额叶皮质突触可塑性的影响.方法 将30只出生后21 d的雄性SD大鼠随机分为去势组、模型组和对照组.去势组和模型组大鼠采用国际通用应激方法建立病理性攻击模型,持续到青春期.采用居住-入侵实验检测青春期大鼠攻击行为,ELISA法检测睾酮水平,蛋白质印迹分析检测前额叶皮质突触后致密物质95(PSD-95)和生长相关蛋白43(GAP-43)表达水平.结果 居住-入侵实验结果显示,去势组攻击潜伏期显著低于模型组和对照组(P＜0.01,P＜o.01),屈服后继续攻击次数高于模型组和对照组(P＜0.01,P＜0.01).ELISA检测结果显示,去势组睾酮水平显著低于模型组和对照组(P＜0.01,P＜0.01),且睾酮水平与攻击各指标呈中-高度负相关(P＜0.01).蛋白质印迹结果显示,去势组前额叶皮质PSD-95和GAP-43表达水平均低于模型组和对照组(P＜0.01,P＜0.01).结论 低水平血清睾酮对青春期大鼠前额叶皮质结构可塑性有损伤,可能与其病理性攻击行为相关.     

FGF21 protects human umbilical vein endothelial cells against high glucose-induced apoptosis via PI3K/Akt/Fox3a signaling pathway

Aims

Diabetic macroangiopathy is the main cause of morbidity and mortality in patients with diabetes. Endothelial cell injury is a pathological precondition for diabetic macroangiopathy. Fibroblast growth factor 21 (FGF21) is a key metabolic regulator which has recently been suggested to protect cardiac myocytes and vascular cells against oxidative stress-induced injury in vitro and vivo. In this study, we aimed to investigate the protective capacity of FGF21 in human umbilical vein endothelial cells (HUVECs) against high glucose (HG)-induced apoptosis via phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt)/FoxO3a pathway.

ALEX1 may be a novel biomarker for human cervical squamous cell carcinoma

The armadillo repeat proteins were first found in armadillo gene of Drosophila. Since then a number of proteins containing armadillo repeats have been noticed and studied. These proteins that consist of 6 to 13 armadillo repeat domains are classified as family of armadillo repeat proteins. Recently, several studies indicated that armadillo repeat family of proteins play an important role in the tumorigenesis and maintenance of tissue integrity. ALEX1 (Arm protein lost in epithelial cancers, on chromosome X), contains two armadillo repeats domains, is expressed different in normal and carcinomas tissues. Several studies have found that ALEX1 protein lost in tumors that originated in epithelial tissues. We evaluated the ALEX1 protein expression in 53 cervical cancers and in 53 non-cancerous cervical tissues from patients and adjacent non-cancerous tissues using immunohistochemistry Results: ALEX1 protein expression is significantly increased in 53 cervical cancers tissues compared with non-cancerous tissues. We found, for the first time, that ALEX1 protein expression in cervical cancers tissues is higher than non-cancerous tissues. It is suggested that the ALEX1 protein is associated with tumorigenesis in cervical cancer and we speculate that the ALEX1 may plays a role as an oncogene in cervical cancer. Moreover, ALEX1 may serve as a novel potential diagnostic biomarker in identifying cervical cancer.