香猪睾丸miR-449b和miR-222分子的发育性变化

为了阐明miR-449b和miR-222对贵州从江香猪睾丸发育的调节作用。运用qPCR方法检测香猪睾丸组织中miR-449b和miR-222表达量的发育性变化,通过miRSystem数据库、Kobas数据库对预测的靶基因进行信号转导通路富集分析。经克隆测序,得到香猪miR-222序列,与猪的相似性为100%,而且物种之间miR-449b和miR-222两个分子的保守性都很高。香猪睾丸中miR-449b于1~4月龄期间的表达量逐渐上升,4月龄时表达量达到峰值,之后下降;miR-222的表达量呈现相反的变化趋势,于1~4月龄期间表达量逐渐下降,1月龄时最高,4月龄时最低,之后的表达量有所回升;已知物种的miR-449b和miR-222序列高度保守;经预测miR-449b靶基因有420个,富集于卵母细胞减数分裂、Notch信号通路、Ras信号通路等100个途径(P0.05),miR-222的靶基因有429个,富集于细胞周期、Fox O信号通路、TGF-β信号通路等69个通路(P0.05);miR-449b和miR-222的靶基因共享的通路为Wnt信号通路、MAPK信号通路、雌激素信号通路等42个(P0.05)。     

麻醉剂MS-222对金鱼体内AKP、CAT和ACP活性的影响

在实验室条件下,采用4个浓度0mg/L、30mg/L、60mg/L和90mg/L,分析了麻醉剂MS-222对金鱼鳃、肝脏和肌肉组织中碱性磷酸酶(AKP)、酸性磷酸酶(ACP)和过氧化氢酶(CAT)活性的影响。试验结果表明:ACP明显出现一个先增后降的趋势,呈抛物线状;在低浓度(≤60mg/L)下,酶活性在各组织中呈现一定的稳定状态,而当麻醉剂浓度升高和麻醉时间增长,酶活性逐渐下降。AKP在低浓度麻醉剂溶液中,酶活性有明显的增加,随着浓度的升高(≥60mg/L)和时间的增长,酶活性呈抑制状态。而在肌肉中这种情况并不明显。CAT在一定阶段(30mg/L～60mg/L)呈现稳定的状态、而后抑制逐渐明显,当麻醉剂MS-222浓度达到一定量(≥60mg/L)以后,对鱼体的刺激就不再增加了,酶活性也相对处于较稳定的状态;当麻醉剂随着时间的增长和浓度的增大时,在鱼体内富集的麻醉剂破坏鱼体内的过氧化氢酶的分子构象,使酶体失去活性。     

Anaesthetic efficacy of MS‐222 and AQUI‐S® in advanced size fry of rohu,Labeo rohita, (Hamilton‐Buchanan)

Anaesthetic agents are very useful for reducing the stress caused by handling, sorting, transportation, artificial reproduction, tagging, administration of vaccines and surgical procedures in fish. The efficacy of two anaesthetics: MS‐222 and AQUI‐S^® were tested on rohu, Labeo rohita advanced size fry. The lowest effective doses that produced induction in 3 min or less and recovery times 5 min or less and meet the most criteria of good anaesthetic characteristics were 125 mg L^?1 of MS‐222, and 30 mg L^?1 of AQUI‐S^® in rohu, Labeo rohita advanced size fry. Induction times were significantly decreased with increased in the concentrations of any of the two tested anaesthetic agents. The lowest doses suitable for transportation of rohu advanced size fry observed were: 10–15 mg L^?1 of MS‐222 and 2.5 mg L^?1 AQUI‐S^®. Both anaesthetics showed promising to be used as anaesthetics for handling and transportation in rohu (Labeo rohita) advanced fry.     

黑色素细胞中过量表达miR-137对TYRP-1和TYRP-2的影响

【目的】证明miR-137通过调控MITF而间接影响小鼠黑色素细胞TYRP-1和TYRP-2的表达,从而影响黑色素的生成。【方法】通过细胞转染技术在细胞水平过量表达miR-137,对黑色素细胞中黑色素含量进行了测定,经RT-PCR以及western blot检测转染后细胞TYRP-1和TYRP-2在mRNA和蛋白水平的变化。【结果】黑色素含量明显减少,在mRNA水平,TYRP1显著降低至0.4倍,TYRP-2显著降低至0.6倍。在蛋白水平,TYRP-1显著降低至0.61倍,TYRP-2显著降低至0.73倍。【结论】过量表达miR-137会通过调控MITF而使黑色素细胞TYRP-1和TYRP-2的表达量降低,从而揭示了miR-137通过调节TYRP-1和TYRP-2的表达间接调控了黑色素的生成。     

Diagnostic value of serum miR-21 for diabetic nephropathy

AIM:To evaluate the diagnostic value of serum miR-21 as a molecular biomarker in the patients with diabetic nephropathy (DN). METHODS:Twenty-five patients with DN, 50 patients with type II diabetes ［divided into DMA group and DMB group base on urinary microalbumin (UmAlb), 25 in each group］, 25 patients with diabetic nephropathy-induced uremia (UM) and 25 normal controls (NC) were included in the study. Total RNA was extracted from the serum samples for determining the miR-21 level using real-time PCR. The relationship between miR-21 level and clinical parameters of the patients with diabetic nephropathy was analyzed. RESULTS:The relative expression level of serum miR-21 was significantly lower in DN group than that in DMA group, DMB group and NC group, and was significantly higher than that in UM group (all P<001). The serum miR-21 level showed significant negative correlation with cystatin C, UmAlb, UmAlb/urinary creatinine in DN patients (P<0.01,P<0.05,P<0.05). For discriminating the DN patients from NC, DMA, DMB, DMA＋NC, DMB＋NC and DMA＋DMB, the areas under the receiver operating characteristic curve (AUC-ROC) for serum miR-21 were 0848 (95% CI: 0737~0959), 0896 (95% CI: 0812~0980), 0782 (95% CI: 0641~0922), 0838 (95% CI: 0743~0933), 0796 (95% CI: 0675~0917) and 0808 (95% CI: 0704~0911), and the sensitivity and specificity were 800% and 720%, 720% and 880%, 720% and 840%, 760% and 770%, 760% and 820%, 700% and 860%, respectively. For discriminating the DN patients from DMA＋DMB＋NC, the AUC-ROC was 0845 (95% CI: 0752~0939), and the sensitivity and specificity were 760% and 773%, respectively. CONCLUSION: Serum miR-21 can serve as a molecular diagnostic marker for diagnosis of diabetic nephropathy.     

miR-144表达质粒的构建及其对肝癌细胞生物学行为的影响

【目的】构建microRNA-144(miR-144)的真核表达质粒,并探讨miR-144对人肝癌细胞株HepG2生物学行为的影响,为肝癌的生物学治疗提供依据。【方法】构建miR-144真核表达质粒并合成miR-144反义寡核苷酸,通过实时定量PCR检测miR-144的表达,采用MTT比色法及流式细胞仪技术,检测抑制与过表达miR-144对肝癌细胞增殖、凋亡和细胞周期的影响。【结果】成功构建了miR-144的真核表达质粒,过表达miR-144能够显著抑制肝癌细胞的增殖(P<0.05),促进细胞凋亡(P<0.05);而抑制miR-144的表达则得到相反的结果;抑制与过表达miR-144对肝癌细胞周期的影响都不大(P>0.05)。【结论】miR-144能影响人肝癌细胞株HepG2的生物学行为,具有抑制肝癌细胞增殖的作用。     

低氧胁迫下鲢miR-17a-5p及其靶基因分析

为了探究miR-17a-5p在鲢低氧胁迫下的功能,在前期鲢small RNA测序的基础上对鲢miR-17a-5p进行靶基因预测及功能富集分析,通过双荧光素酶活性验证其与HIF-1α的靶向关系,并检测低氧胁迫下miR-17a-5p和其靶基因在鲢肝脏、脑、心脏和鳃四个组织中的动态表达特征。结果显示,鲢miR-17a-5p在不同物种间高度保守,预测出381个miR-17a-5p的潜在靶基因显著富集在硫代谢、mTOR信号通路以及萜类骨架的生物合成3个KEGG信号通路上。miR-17a-5p可与HIF-1α mRNA的3′UTR结合,并降低HIF-1α mRNA水平,低氧胁迫下miR-17a-5p的表达呈下降趋势,而HIF-1α表达则呈上升趋势;3个显著富集KEGG通路中的11个miR-17a-5p潜在靶基因在低氧胁迫过程中的不同组织中的表达,尤其是肝脏中的表达,除SGK1外,均呈显著上升趋势。研究表明,低氧胁迫下鲢各组织中miR-17a-5p的表达下调减弱了其对靶基因的抑制作用,进而导致HIF-1α、ddit4和Lrp5等响应低氧胁迫的基因表达上调。本研究为低氧胁迫下鲢miRNA的表达与调控机...     

miR-21:生成、表达调控及对其细胞凋亡的影响

凋亡是细胞的一种基本生物学现象,在机体生命过程中发挥着重要作用。miRNAs是在真核生物内发现的一类长度约22个核苷酸的内源性非编码单链小RNA,通过调控靶基因的表达参与调控细胞的凋亡、分化、增殖、侵袭和代谢。大量的研究表明,miR-21作为miRNAs重要成员之一,与细胞凋亡有重要联系。本文将从miR-21的生成、表达调控及对细胞凋亡的影响方面进行综述。     

猪miR-206前体的克隆、表达及生物信息学分析

研究骨骼肌中表达的miRNAs对深入探明骨骼肌生长发育的调控机制具有十分重要的意义。采用比较基因组学与生物信息学相结合的方法,从猪骨骼肌中克隆了猪miR-206的前体（Precursor）,对该miRNA前体的二级结构及与其它物种序列的同源性进行了分析。RT-PCR结果显示,该前体miRNA在猪大多数组织中均有表达,在骨骼肌中表达量最高。生物信息学分析表明miR-206前体序列在不同物种间具有较高的保守性。     

斑马鱼miR-30e对血细胞生成的作用机制研究

microRNA(简写为miRNA)是一类内源基因编码的单链RNA分子,参与转录后基因的表达调控,在血液生成过程中起着重要作用.为研究miR-30e对鱼类血液生成的作用机制,以斑马鱼Danio rerio作为模式生物,针对青藏高原裂腹鱼血液组织中高表达的miR-30e,利用生物信息软件预测斑马鱼miR-30e的靶基因,...