结直肠癌组织NDRG2和CA19-9表达水平与预后的相关性分析

目的分析结直肠癌(CRC)组织N-Myc下游调节基因2(NDRG2)和糖链抗原19-9(CA19-9)表达与患者预后的相关性。方法选择2013年5月至2014年5月本院接收的66例CRC患者为研究对象,用免疫组化法测定患者术中所得肿瘤组织和癌旁正常组织标本中NDRG2和CA19-9的表达情况,并分析其与患者病理特征和预后的关系。结果癌变组织NDRG2阳性表达率显著低于癌旁正常组织,CA19-9阳性表达率显著高于癌旁正常组织(P<0.05)。肿瘤病灶Dukes分期晚、分化程度低、淋巴或肝转移者NDRG2阳性占比均低于Dukes分期早、分化程度中高、非淋巴或肝转移者,CA19-9阳性占比高于Dukes分期早、分化程度中高、非淋巴或肝转移者(P<0.05)。NDRG2阳性者和CA19-9阴性者累计生存时间明显长于NDRG2阴性者和CA19-9阳性者(P<0.05)。结论 CRC瘤体中NDRG2低表达,CA19-9高表达,且其表达与患者病理分期、分化程度、淋巴或肝脏是否转移有关。     

丁酸钠对食管癌细胞增殖的影响

目的:观察丁酸钠对食管癌EC9706细胞增殖、细胞周期及NDRG1蛋白表达的影响.方法:采用0.5 mmol/L丁酸钠作用于食管癌EC9706细胞,用MTT法检测处理不同时间时细胞增殖的变化,采用流式细胞仪检测细胞周期的变化,采用免疫组化方法检测NDRG1蛋白表达的变化.结果:①MTT实验结果显示,丁酸钠作用1～5 d时各组吸光度均低于对照组,经统计学处理差异有显著性意义(P＜0.05);丁酸钠不同作用时间的细胞增殖抑制率分别为:1 d组21.2%,3 d组23.5%,5 d组25.6%,且随作用时间延长抑制率升高.②细胞周期检测结果为(%):G1期细胞:对照组57.00±1.10,1 d组63.10±0.78,3 d组67.20±0.58,5 d组69.70±0.64;S期细胞:对照组25.30±0.27,1 d组20.20±0.66,3 d组20.40±0.82,5 d组20.90±0.50;M期细胞:对照组17.20±1.00,1 d组16.70±0.69,3 d组12.10±0.41,5 d组9.40±0.23.各组G1期结果分别与对照组结果比较,差异有显著性意义(P＜0.01).③丁酸钠可上调NDRG1蛋白表达水平,随作用时间增长蛋白表达增强.结论:丁酸钠可抑制食管癌细胞增殖,这一作用可能与NDRG1蛋白表达增加有关.     

NDRG3对前列腺癌细胞致癌潜能的影响

目的：研究过表达外源性和RNA干扰内源性NDRG3对前列腺癌细胞克隆形成的影响。方法：将NDRG3表达质粒稳定转染人类PC3前列腺癌细胞株,用1个NDRG3稳定表达亚克隆与做为对照的亲代和空质粒转染的PC3细胞进行克隆形成实验;同时利用RNA干扰技术沉默CL-1细胞的内源性NDRG3,并用Western blotting技术进行鉴定,在对照组、GFP-siRNA组和NDRG3-siRNA组进行克隆形成实验。结果：过表达外源性NDRG3的PC3细胞形成的克隆数明显多于亲代和空质粒转染的PC3细胞株;在RNA干扰实验中,CL-1对照组和GFP-siRNA组形成的总克隆数和大克隆数均明显多于NDRG3-siRNA组（P<0.05）。结论：NDRG3可以提高前列腺癌细胞的致癌潜能和克隆形成能力。     

食管鳞癌组织中NDRGl mRNA和蛋白的表达

目的：探讨食管鳞癌组织中NDRGlmRNA和蛋白的表达情况及其与临床病理因素的关系。方法：分别采用RT-PCR和免疫组织化学SP法检测49例食管鳞癌组织（分化程度Ⅰ级14例，Ⅱ级23例，Ⅲ级12例；有淋巴结转移2l例，无淋巴结转移28例）、23例癌旁不典型增生黏膜组织及49例正常食管黏膜组织中NDRG1 mRNA和蛋白的表达。结果：食管鳞癌组织中NDRG1 mRNA相对含量低于癌旁不典型增生黏膜组织及正常食管黏膜组织（P〈0．05或0．01）；不同分化程度的食管鳞癌组织之间NDRG1 mRNA相对含量差异无统计学意义（P〉0．05）；伴淋巴结转移的食管鳞癌组织与无淋巴结转移的食管鳞癌组织NDRG1 mRNA相对含量差异无统计学意义（P〉0．05）。癌旁不典型增生组织及食管鳞癌组织中NDRG1蛋白（＋＋）阳性率（3／23和4／49）均低于正常黏膜组织的43／49（P〈0．01）；食管鳞癌组织中NDRG1蛋白表达与癌组织的分级及有、无淋巴结转移无关（P均〉0．05）。结论：食管鳞癌组织中NDRG1 mRNA和蛋白表达降低，其低表达可能与食管鳞癌发生有关。     

丁酸钠对食管癌细胞NDRG1基因表达的影响

目的了解丁酸钠对食管癌细胞分化相关基因NDRG1表达的影响。方法采用丁酸钠作用于食管癌EC9706细胞，应用原位杂交法和免疫组化SP法检测丁酸钠作用1、3d及5d后NDRG1蛋白和mRNA的表达。结果①丁酸钠作用1～5dNDRG1mRNA表达灰度值，对照组86±15，1d组125±14，3d组142±25，5d组164±21，1、3、5d组与对照组比较，差异均有统计学意义（均P〈0．01）；②丁酸钠作用1—5dNDRG1蛋白表达灰度值，对照组145±18，1d组179±24，3d组188±20，5d组193±26，1、3、5d组与对照组比较，差异均有统计学意义（均P〈0．01）。结论丁酸钠可上调EC9706细胞分化相关基因NDRG1表达。促使其分化。     

Ethanol and PTZ effects on siRNA‐mediated GABAB1 receptor: Down regulation of intracellular signaling pathway in prenatal rat cortical and hippocampal neurons

GABA_B receptors (R) are widely expressed and distributed in the nervous system, and have been implicated in variety of neurodegenerative and pathophysiological disorders. However, the exact molecular mechanism regarding responsibility of GABA_B1R in downstream signaling pathway is not well understood. The present study was undertaken to explore the downstream signaling and role of GABA_B1R upon acute ethanol and pentylenetetrazol (PTZ) exposure for (20 min) in cortical and hippocampal neuronal cell cultures by using GABA_B1R RNA interference (i) (30 nM, 48 h) at gestational days 17.5. The results showed that GABA_B1R and protein kinase A‐α (PKA) showed decreased expression upon ethanol and PTZ exposure in cortical and hippocampal neurons during transfected and nontransfected conditions, whereas these effects could lead to significant changes in phosphorylation of cAMP‐response element binding protein (p‐CREB) expression where GABA_B1R was knocked down. Furthermore, intracellular Ca⁺² concentrations were also reduced in some groups after transfection with GABA_B1R RNAi. These results showed a critical role of GABA_B1R upon ethanol and PTZ exposure by modulating downstream signaling pathway. Finally, these findings suggested that inhibition of GABA_B1R results in the modulation of PKA, p‐CREB pathway may play a role in long‐term changes in the nervous system, and may be an underlying cause of ethanol's effects. Synapse 64:181–190, 2010. © 2009 Wiley‐Liss, Inc.     

三叶因子2和N-myc下游调节基因1在不同子宫内膜组织中的表达

目的探讨三叶因子2(TFF2)和N-myc下游调节基因1(NDRG1)在不同子宫内膜组织中的表达,为早期诊断子宫内膜癌提供新的生物学指标。方法收集珠海市人民医院经手术切除并经病理检查确诊为子宫内膜癌组织157例,另选择同期经手术切除并经病理检查确诊的子宫内膜不典型增生组织30例及其他病因手术切除的正常子宫内膜20例,分别检测其TFF2和NDRG1蛋白的表达情况,并分析TFF2和NDRG1蛋白的表达与子宫内膜癌临床病理因素的关系。结果与正常子宫内膜和子宫内膜不典型增生组织比较,TFF2在子宫内膜癌组织中的阳性表达率显著降低(P<0.05),但NDRG1在子宫内膜癌组织中的阳性表达率显著升高(P<0.01)。TFF2在Ⅰ、Ⅱ期子宫内膜癌组织中的阳性表达率显著高于Ⅲ^Ⅳ期(P<0.05);高、中分化子宫内膜癌组织中TFF2的阳性表达率显著高于低分化组织及其他类型(P<0.01)。与深肌层浸润的子宫内膜癌比较,浅肌层浸润的子宫内膜癌组织中TFF2的阳性表达率显著升高(P<0.05);与有淋巴结转移的子宫内膜癌比较,无淋巴结转移的子宫内膜癌组织中TFF2的阳性表达率显著升高(P<0.01)。而高、中度分化的子宫内膜癌组织中NDRG1的阳性表达率显著低于低分化及其他类型(P<0.05)。浅肌层浸润的子宫内膜癌组织中NDRG1的阳性表达率显著低于深肌层浸润(P<0.01),与无淋巴结转移的子宫内膜癌比较,有淋巴结转移的子宫内膜癌组织中NDRG1的阳性表达率显著升高(P<0.01)。结论在子宫内膜组织中,TFF2表达的缺失及NDRG1蛋白的高表达可能与子宫内膜癌的发生发展有重要关系,有望在子宫内膜癌的早期诊断、判断是否存在侵袭转移等临床评估中发挥作用。     

Octopamine-mediated circuit mechanism underlying controlled appetite for palatable food in Drosophila

The easy accessibility of energy-rich palatable food makes it difficult to resist food temptation. Drosophila larvae are surrounded by sugar-rich food most of their lives, raising the question of how these animals modulate food-seeking behaviors in tune with physiological needs. Here we describe a circuit mechanism defined by neurons expressing tdc2-Gal4 (a tyrosine decarboxylase 2 promoter-directed driver) that selectively drives a distinct foraging strategy in food-deprived larvae. Stimulation of this otherwise functionally latent circuit in tdc2-Gal4 neurons was sufficient to induce exuberant feeding of liquid food in fed animals, whereas targeted lesions in a small subset of tdc2-Gal4 neurons in the subesophageal ganglion blocked hunger-driven increases in the feeding response. Furthermore, regulation of feeding rate enhancement by tdc2-Gal4 neurons requires a novel signaling mechanism involving the VEGF2-like receptor, octopamine, and its receptor. Our findings provide fresh insight for the neurobiology and evolution of appetitive motivation.The adaptive control of foraging decisions is crucial to survival and reproduction and is mediated by complex brain mechanisms. For example, in hungry animals, feeding behaviors can be modulated by diverse neural systems including those responsible for receiving and processing sensory properties and assigning reward and motivational significance of food stimuli (1–3). At present, elucidation of molecular and circuit mechanisms underlying the adaptive control of feeding behavior remains highly challenging.Our previous studies have shown that Drosophila larvae, like mammals, display diverse adaptive foraging strategies in response to appetizing odors or satiety state and food quality (4–6). For example, larvae fed for ad libitum intake tend to prefer soft, liquid sugar media that contain readily ingestible sugar solution but decline solid media in which sugar solution is embedded in gelled agar and is less accessible (5). However, as food deprivation is prolonged, larvae will become increasingly persistent in extracting the sugar solution from solid media (7). We have also shown that an evolutionarily conserved signaling cascade, involving neuropeptide F (NPF, the fly homolog of neuropeptide Y, or NPY) and insulin-like peptides (dILPs), selectively integrates motivational state (hunger) with persistence to pulverize solid food (5, 7).The observation that the conserved NPY-like system selectively promotes food acquisition behaviors that require high energetic cost has led us to postulate that fly larvae may use other conserved neural mechanisms to regulate acquisition of readily accessible palatable food. In this work, we provide evidence which supports this hypothesis. We show that an octopamine (OA)/β-adrenergic-like receptor (Octß3R)-dependent circuit mechanism selectively regulates appetite for soft sugar media. This circuit mechanism seems to involve two subsets of tdc2-Gal4 neurons in the subesophogeal ganglia (SOG). One of them mediates the hunger-driven increase of feeding and is modulated by a novel activity of the VEGF2-like receptor pathway (8). The other is required for preventing excessive appetite in fed larvae. This and our previous findings provide fresh mechanistic insights into how brain mechanisms differentially organize appetitive motivations in responses to high- and low-quality food sources under different energy states.