糖皮质激素受体基因(NR3C1)在肾透明细胞癌中的表达差异及意义

目的通过数据库分析糖皮质激素受体基因(NR3C1)在肾透明细胞癌中的表达差异及意义。方法利用Oncomine数据库检索并筛选出NR3C1的信息,得到NR3C1在ccRCC与正常肾组织中的表达情况,使用GEPIA数据库分析NR3C1的表达与ccRCC患者在TCGA数据中生存率之间的关系。通过String数据库分析NR3C1基因的蛋白相互作用及上下游调控关系。结果NR3C1在ccRCC组织及正常组织中表达,差异有统计学意义(P<0.05);高表达NR3C1的患者总体生存率显著高于低表达NR3C1的患者,差异有统计学意义(P<0.05);同时,NR3C1与NCOA1、NCOA2、NOCRO1、HSP90AA1、FKBP5等蛋白存在相互作用,并且参与了炎症反应,细胞增殖和分化等生物过程。结论NR3C1在ccRCC中显著高表达,并与ccRCC的预后相关,可作为诊断和治疗的潜在靶点。

(-)-Epigallocatechin-3-gallate enhances poly I:C-induced interferon-λ1 production and inhibits hepatitis C virus replication in hepatocytes

AIM To investigate the effect of(-)-epigallocatechin-3-gallate(EGCG) on polyinosinic-polycytidylic acid(poly I:C)-triggered intracellular innate immunity against hepatitis C virus(HCV) in hepatocytes. METHODS A cell culture model of HCV infection was generated by infecting a hepatoma cell line, Huh7, with HCV JFH-1 strain(JFH-1-Huh7). Poly I:C with a high molecular weight and EGCG were used to stimulate the JFH-1-Huh7 cells. Real-time reverse transcription-polymerase chain reaction was used to detect the expression levels of intracellular m RNAs and of intracellular and extracellular HCV RNA. Enzyme-linked immunosorbent assay was used to evaluate the interferon(IFN)-λ1 protein level in the cell culture supernatant. Immunostaining was used to examine HCV core protein expression in Huh7 cells.RESULTS Our recent study showed that HCV replication could impair poly I:C-triggered intracellular innate immune responses in hepatocytes. In the current study, we showed that EGCG treatment significantly increased the poly I:C-induced expression of Toll-like receptor 3(TLR3), retinoic acid-inducible gene I, and IFN-λ1 in JFH-1-Huh7 cells. In addition, supplementation with EGCG increased the poly I:C-mediated antiviral activity in JFH-1-Huh7 cells at the intracellular and extracellular HCV RNA and protein levels. Further investigation of the mechanisms showed that EGCG treatment significantly enhanced the poly I:C-induced expression of IFN-regulatory factor 9 and several antiviral IFNstimulated genes, including ISG15, ISG56, myxovirus resistance A, and 2'-5'-oligoadenylate synthetase 1, which encode the key antiviral elements in the IFN signaling pathway. CONCLUSION Our observations provide experimental evidence that EGCG has the ability to enhance poly I:C-induced intracellular antiviral innate immunity against HCV replication in hepatocytes.

Metabolic and Endocrine Responses of Desert-Adapted Mice Reproductive System to Increased Salinity

From an evolutionary point of view, reproduction timing is an important adaptation which enables the transfer of genetic properties, thus enabling species continuation. Rodents inhabiting arid environments need reliable cues for triggering their reproduction. Results of previous studies showed that increased dietary salinity plays an important role as an ultimate regulator for desert adapted rodents＇ reproductive system. The authors aimed discovering pathways by which high salinity can affect the reproductive system and metabolic status of desert adapted common spiny mice, Acomys cahirinus. Mice were challenged with osmotic stress, water source salinity increased gradually from 0.9% - 5% NaCI under short days （SD） and long days （LD）. The authors assessed leptin and free fatty acid （FFA） levels using ELISA while, SYBR green technology was used for relative receptor expression （RQ） of target genes. Results revealed that serum levels of the hormone leptin were significantly （P 〈 0.05） reduced in salinity treated （ST） mice. Levels of FFA were significantly （P 〈 0.05） increased in LD- and SD-ST-males. In ST-SD females a significant increase （P 〈 0.05） in expression levels of leptin （Ob-Rt） mRNA receptor gene, in ovaries was noted. Aldosteron （Nr3c2） and vasopressin （AVP） mRNA receptor expression genes levels were significantly （P 〈 0.05） increased in both LD- and SD- ST- males.

雷帕霉素对衰老细胞生物节律关键基因的调控作用观察

目的观察雷帕霉素对衰老细胞生物节律关键基因的调控作用。方法通过传代诱导人皮肤成纤维细胞复制性衰老,获得衰老的人皮肤成纤维细胞。加入50%的马血清刺激诱导细胞同步化,将细胞分为观察组和对照组,分别加入100 mmol/L的雷帕霉素和DMSO培养24 h。采用荧光定量PCR法检测生物节律关键基因芳烃受体核转位蛋白3(BMAL1)、节律周期蛋白2(PER2)、蓝光受体蛋白1(CRY1)、孤儿核受体α(Rev-erbα)、孤核受体α(Rorα)mRNA表达,利用JTK-CYCLE软件分析各基因的节律周期,Western blotting法检测BMAL1、PER2、CRY1蛋白表达。结果观察组PER2、CRY1、Rev-erbα、RorαmRNA表达高于对照组(P均<0.05)。对照组BMAL1、CRY1、Rev-erbα、Rorα基因的节律周期为24 h,PER2基因的节律周期为28 h。观察组BMAL1、CRY1、Rorα基因的节律周期为24 h,Rev-erbα基因的节律周期为27.5 h,PER2的节律周期为24 h。观察组CRY1蛋白表达高于对照组(P<0.05),PER2蛋白表达低于对照组(P<0.05),两组BMAL1 mRNA及蛋白表达比较无统计学差异。结论雷帕霉素可以提高衰老的人皮肤成纤维细胞生物节律关键基因BMAL1、PER2、CRY1、Rev-erbα、RorαmRNA的表达,缩短PER2的节律周期,对紊乱的节律钟功能具有调控作用。

儿童哮喘NR3C1基因单核苷酸多态性及对糖皮质激素疗效的影响分析

目的探讨糖皮质激素受体基因(NR3C1)单核苷酸多态性与儿童哮喘的关系及吸入性糖皮质激素(inhaled corticosteroid,ICS)疗效的分析。方法选取2018年10月至2020年10月于北部战区总医院住院治疗的儿童支气管哮喘患儿101例为哮喘组,100例健康体检儿童为对照组,提取所有入选儿童外周血DNA,采用SNaPshot SNP基因检测技术对糖皮质激素受体基因位点进行多态性分析,比较两组儿童rs41423247、rs7701443位点基因型频率及等位基因频率,对哮喘组患儿予ICS治疗12周时,比较不同基因型组哮喘治疗效果差异。结果rs41423247位点基因型GG/GC/CC频率分别为:哮喘组75.2%、21.8%、3.0%,对照组72.0%、24.0%、4.0%,两组比较差异无统计学意义(χ^(2)=0.333,P>0.05);rs7701443位点基因型GG/GA/AA频率分别为:哮喘组45.5%、39.6%、14.9%,对照组56.0%、31.0%、13.0%,两组比较差异无统计学意义(χ^(2)=2.259,P>0.05)。经ICS治疗后,rs41423247位点CC型哮喘患儿C-ACT/ACT评分较治疗前无显著升高(P>0.05),而GG型、GC型哮喘患儿的C-ACT/ACT评分较治疗前均显著升高(P<0.05);GG型与GC型、CC型哮喘患儿治疗后的C-ACT/ACT评分比较差异均有统计学意义(P<0.05)。rs7701443位点3种基因型哮喘患儿C-ACT/ACT评分较治疗前均有显著升高(P<0.05),但治疗后3种基因型的C-ACT/ACT评分差异无统计学意义(P>0.05)。经ICS治疗后,rs41423247位点GG型、GC型哮喘患儿的各项肺功能指标较治疗前均有显著改善(P<0.05),而CC型哮喘患儿只有MMEF较治疗前有显著改善(P<0.05);经ICS治疗后,rs7701443位点3种基因型各项肺功能指标均较治疗前有显著改善(P<0.05)。结论NR3C1位点rs41423247、rs7701443均存在基因多态性,该研究未发现儿童哮喘患者与健康对照者中NR3C1位点rs41423247、rs7701443的基因多态性存在差异。儿童哮喘患者NR3C1位点rs41423247、rs7701443的不同基因型频率对ICS治疗效果存在差异。