胃癌组织中P16、Cyclin D1、Rb表达及其生物学行为的研究

目的研究P16、Cyclin、D1Rb基因在胃癌中的表达及其与胃癌生物学行为的关系.方法采用免疫组化方法检测10例正常胃黏膜、30例胃癌组织中P16、Cyclin D1、Rb蛋白的表达,并结合其临床资料进行分析.结果P16、Cyclin D1、Rb蛋白阳性表达,正常胃粘膜分别为90%、10%、90%;胃癌分别为36.67%、53.33%、50%,胃癌纽与正常对照组间差异均有显著性(P＜0.05),P16、Cyclin D1蛋白与胃癌分化程度、淋巴结转移与否、远处转移与否呈一定的相关性.结论P16、Cyclin D1、Rb基因与胃癌发生有关,P16、Cyclinp蛋白检测有助于胃癌预后的判断.     

Rb1基因第16内含子内21个碱基缺失1例

目地研究双眼视网膜母细胞瘤患者Rb1基因杂合性突变的分子生物学特性。方法应用PCR—SSCP直接测序技术检测双眼视网膜母细胞瘤患者白细胞DNA中Rb1基因杂合性突变。结果50例证实有Rb1基因杂合性突变的病例中有1例发生于第16内含子中可以用3种定位方法解释、具有相同序列的21个碱基缺失。结论这种极为少见的Rb1基因突变方式可能是由于破坏了正常拼接位点的结构而激活了“隐蔽拼接位点”，导致异常的Rb1基因mRNA产生或由此影响整个拼接过程。     

脑络康缓释胶囊的制备工艺和体外释放度的研究

以羟丙甲基纤维素作为主要的缓释辅料,采用正交设计方法优化处方研制成脑络康缓释胶囊.以人参皂苷的累积释放量作为考察指标,按照<药典>规定的缓释制剂的释放度进行优选.结果:最佳的处方工艺即羟丙甲基纤维素216 mg,乙基纤维素24 mg,混合稀释剂132 mg,释放曲线符合Higuchi动力学模型.     

人参皂苷Rg1诱导骨髓间充质干细胞分化为神经样细胞的作用

目的研究人参皂苷Rg1在体外能否诱导Wistar大鼠骨髓间充质干细胞分化为神经元样细胞。方法通过贴壁法分离大鼠骨髓间充质干细胞,体外培养扩增,人参皂苷Rg1诱导分化,光镜下观察细胞形态,免疫细胞化学检测神经元特异性烯醇化酶（NSE）和神经胶质纤维酸性蛋白（GFAP）的表达,RT-PCR检测细胞NGF mRNA的表达。结果大鼠骨髓间充质干细胞可通过贴壁法成功分离并可以在体外大量扩增。人参皂苷Rg1诱导72h后,部分骨髓间充质干细胞（35．57％±3．59％）转变为神经元样细胞,免疫细胞化学染色NSE呈阳性,分化的神经元样细胞可能表达NGF mRNA。结论人参皂苷Rg1可以在体外诱导大鼠骨髓间充质干细胞分化为神经元样细胞,并且可能表达NGF mRNA。     

西洋参和胡萝卜体细胞融合培养杂种鉴定及愈伤组织中人参皂苷Rb_1含量分析

目的 :通过采用体细胞融合技术将西洋参基因转入胡萝卜中 ,为贵重中药、生长受地理环境等限制的中药扩大药源 ,利用杂种优势为培养适于大面积栽培且有效成分人参皂苷含量较高的优良杂交品种提供理论与实验依据。方法 :用PEG法对五加科植物西洋参与伞形科植物胡萝卜进行体细胞融合 ,通过同工酶进行初步杂种鉴定并用HPLC法测定西洋参和胡萝卜体细胞融合培养愈伤组织中人参皂苷Rb1含量。结果 :体细胞融合技术成功地获得了西洋参和胡萝卜体细胞杂交愈伤组织 ;同工酶分析是鉴定杂种的有效手段之一 ;在 10个杂交体愈伤组织中有 6个杂交体愈伤组织人参皂苷Rb1含量比未融合前西洋参愈伤组织中的含量高。     

人参皂甙单体Rb1对大鼠在体心脏收缩性能的影响

本文作者为进一步探讨人参皂甙单体Rb1对心肌收缩的影响,完善人参对心肌保护作用的系统性研究,拟在在体工作鼠心上,以心肌收缩性能为指标,对比观察Rb1对心肌收缩的影响.     

The role of Na+/K+ ATPase activity during low flow ischemia in preventing myocardial injury: A 31P, 23Na and 87Rb NMR spectroscopic study

An increase in intracellular Na⁺ during ischaemia has been associated with myocardial injury. In this study, we determined whether inhibition of Na⁺/K⁺ ATPase activity contributes to this increase and whether Na⁺/K⁺ ATPase activity can be maintained by provision of glucose to perfused rat hearts during low flow, 0.5 ml/min, ischemia. We used ³¹P NMR spectroscopy to determine changes in myocardial energetics and intracellular and extracellular volumes. ²³Na NMR spectroscopy, with DyTTHA^3- present as a shift reagent, was used to measure changes in intracellular Na⁺ and ⁸⁷Rb NMR spectroscopy was used to estimate Na⁺/K⁺ ATPase activity from Rb⁺ influx rates, Rb⁺ being an NMR-sensitive congener of K⁺. In hearts provided with 11 mM glucose throughout ischemia, glycolysis continued and ATP was twofold higher than in hearts without glucose. In the glucose-hearts, Rb⁺ influx rate was threefold higher, intracellular Na⁺ was fivefold lower at the end of ischemia and functional recovery during reperfusion was twofold higher. We propose that continuation of glycolysis throughout low flow ischemia allowed maintenance of sufficient Na⁺/K⁺ ATPase activity to prevent the increase in intracellular Na⁺ that would otherwise have led to myocardial injury.     

Rb基因在鼻咽癌中存在状态的研究

首次应用生物素－１４－ｄＵＴＰ标记Ｒｂ３．８ｋｂ探针，结合亲合素－碱性磷酸酶发光及自显影法，对Ｒｂ基因在鼻咽癌中存在状态进行了研究。杂交结果表明，３例正常胎儿鼻咽组织出现分子量为１２．８、１０．２、８．０、６．２、５．６、５．２及４．８ｋｂ的７条杂文区带，１１例鼻咽癌组织均发现有Ｒｂ基因缺失或失活，其中４例为５．６ｋｂ片段缺失，４例为４．８ｋｂ缺失并有２例伴５．６ｋｂ减弱，２例为１０．２ｋｂ缺失，１例为５．６及５．２ｋｂ片段显著减弱，说明在上述１１例鼻咽癌中均有Ｒｂ基因的改变。这种高频率的异常变化，提示Ｒｂ基因的缺失或失活，与鼻咽癌的发生有密切关系。     

Accumulation of p53 in response to adenovirus early region 1A sensitizes human cells to tumor necrosis factor alpha-induced apoptosis

Many tumor cells are resistant to tumor necrosis factor alpha (TNFalpha)-induced apoptosis. Adenovirus early region 1A (AdE1A) sensitizes the otherwise resistant cells to TNFalpha. AdE1A also stabilizes the p53 protein. The present study demonstrates a correlation between AdE1A-induced sensitization and stabilization of p53 in TNFalpha-induced apoptosis since the N-terminal and CR2 regions, the binding sites for CBP/p300, Rb and 26S proteasome regulatory components, are required for both these actions of AdE1A. TNFalpha does not induce apoptosis and AdE1A fails to sensitize TNFalpha cytotoxicity in p53-negative cells. However, introduction of exogenous p53 overcomes the cellular resistance to TNFalpha toxicity and enhances AdE1A sensitization, demonstrating that AdE1A sensitizes TNFalpha-induced apoptosis by its stabilization of p53. A proteasome inhibitor, lactacystin, enhances TNFalpha cytotoxicity in p53-positive and -negative cells, suggesting that accumulation of cellular proteins other than p53 might also regulate the cellular response to TNFalpha signaling.