肝细胞癌患者肝组织中JAB1和P27kip1的表达变化及其临床意义

目的 研究肝细胞癌（HCC）组织中c-Jun激活域连接蛋白1（c-Jun activation domain binding protein 1，JAB1）的表达及与P27kip1(P27)间的关系，探讨JAB1与临床病理及预后的关系。方法 免疫组化检测HCC及 相应癌旁肝组织中JAB1和P27的表达。Western blot及免疫沉淀检测JAB1和P27的表达及相互作用。结果 HCC中，JAB1的表达高于癌旁肝组织，P27表达低于癌旁组织。JAB1的表达与组织分化程度、AFP值及转移相关（P<0.05）。HCC中JAB1与P27的表达呈负相关(r=－0.7077, P<0.001)，并且能免疫共沉淀。结论 JAB1的表达与P27负相关，JAB1作为P27的负性调控因子，可能与HCC的恶性进展及预后相关。     

Substance P potentiates 5-HT3 receptor-mediated current in rat trigeminal ganglion neurons

The present study aimed to investigate the interaction between the coexistent SP receptor and 5-HT3 receptor in trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The majority of the neurons examined responded to 5-HT with an inward current (I5-HT) (78.2%, 79/101) that could be blocked by 5-HT3 receptor antagonist, ICS-205,930. The I5-HT was potentiated by preapplication of SP (10(-10) to 10(-8) M) in most 5-HT-sensitive cells(78.5%, 62/79). Coapplication of SP and GR-82334, antagonist of NK1 receptor, had no enhancing effect on I5-HT. The concentration-response curves for 5-HT with and without SP preapplication show that: (1) the threshold 5-HT concentrations with and without SP preapplication are basically the same, while SP preapplication increased the maximal value of I5-HT by 38.0% of its control; (2) the EC50 values of the curves with and without SP pretreatment are very close, i.e. 1.89 x 10(-5) M and 2.08 x 10(-5) M (P > 0.1; n = 9), respectively. Intracellular dialysis of GDP-beta-S, a non-hydrolyzable GDP analog, and GF-109203X, a selective protein kinase C inhibitor, removed the SP potentiation of I5-HT. These results may offer a clue to understanding the mechanism underlying the generation and/or regulation of peripheral pain caused by tissue damage inflammation, etc.     

Ⅰ型膜型基质金属蛋白酶在癌细胞系中的表达及其与明胶酶A活？…

目的 探讨Ⅰ型膜型基质金属蛋白酶在人肿瘤细胞系中的表达情况及其与明胶酶的分泌和活化的关系。方法 运用逆转录巢式ＰＣＲ,ＲＮＡ印迹分析和蛋白质印迹分析的方法,检测ＭＴ１－ＭＭＰ,ｍＲＮ和ＭＭＰ２蛋白在４个黑色素瘤,２个肺癌和２个前列腺癌细胞系中的表达水平。结果 ＭＴ１－ＭＭＰｍＲＮＡ在所有癌细胞系中均有表达,但表达水平很不一致;在４个黑色素瘤细胞系中的表达水平明显高于肺癌和前列腺癌细胞系;     

细胞周期蛋白E对乳腺癌细胞MCF-7生长及周期相关基因的影响

观察细胞周期蛋白（ｃｙｃｌｉｎ）Ｅ的表达对乳腺癌细胞ＭＣＦ－７生长及其他周期相关基因的影响。方法构建正义和反义ｃｙｃｌｉｎ Ｅ ｃＤＮＡ真核表达载体,并采用ｌｉｐｏｆｅｃｔ ＡＭＩＮＥ转染,将其导入ＭＣＦ－７细胞,获得稳定表达正义及反义ｃｙｃｌｉｎＥ的细胞系,经Ｓｏｕｔｈｅｒｎ和Ｗｅｓｔｅｒｎ ｂｌｏｔ检测有外源性片片段的插入及表达,并作细胞生长曲线、四甲偶氮唑盐（ＭＴＴ）法和流式细胞计数分析;用     

人TH基因重组腺病毒的构建及生物学活性研究

将人酪氨酸羟化酶cDNA表达盒克隆于质粒型腺病毒载体p△Elsp1A，得到重组质粒pAd－TH。随后用脂质体法将重组质粒pAd－TH和拯救型腺病毒质粒pBHG11一起共转染293细胞，通过体内同源重组生成重组腺病毒AdCMVth，THcDNA重组进入腺病毒E1区并受CMV启动子控制。采用形态学、病毒核酸酶切和PCR／RT－PCR等方法进行鉴定正确。重组腺病毒滴度达到1010pfu／ml。初步结果表明，该重组腺病毒感染MN9D细胞后可使细胞内多巴胺水平增加1倍，显示出明显的TH生物学活性。提示TH重组腺病毒AdCMVth可作为高效的基因转移载体用于帕金森氏病基因治疗。     

A组轮状病毒广州地方株VP7基因序列的比较分析

目的 了解我国轮状病毒G1型广州地方株与标准株及北京G1型地方株VP7基因序列的差异，为我国轮状病毒疫苗的研制提供资料。方法 通过逆转录—聚合酶链反应(RT—PCR)获得了轮状病毒广州地方株R97—196 VP7全基因的cDNA片段，将其克隆入T—A克隆质粒pUCm—T中，构建成重组质粒pUCmT—VP7，对克隆的VP7基因进行序列测定。结果 该地方株的基因核苷酸全长为1062nt，读码框架和以往的研究一致，和北京G1型地方株173的VP7氨基酸序列具有高度同源性(98％)，而与不同血清型标准株间则变异较大(73％—81％)，氨基酸序列中存在的一些高变区和保守功能区与已报道的研究结果一致。从进化角度分析，与轮状病毒标准株Wa株，相距较远。结论 轮状病毒广州地方株R97—196 VP7基因片段属G1型，轮状病毒VP7基因的变异与地域有一定关系。     

Effect of intracellular dialysis of ATP on the hyperpolarization-activated cation current in rat dorsal root ganglion neurons

The mechanism of the effect of intracellular ATP on the hyperpolarization-activated non-selective cation current (Ih) in rat dorsal root ganglion neurons was investigated using a whole cell voltage-clamp technique. Under voltage-clamp conditions, Ih was activated by hyperpolarizing pulses raised to a voltage of between -70 and -130 mV. The activation curve of Ih in rat dorsal root ganglion (DRG) neurons shifted by about 15 mV in the positive direction with an intracellular solution containing 1 mM cAMP. When ATP (2 mM) was applied intracellularly, the half-maximal activation voltage (Vhalf) of Ih shifted from -97.4 +/- 1.9 to -86.8 +/- 1.6 mV, resulting in an increase in the current amplitude of Ih by the pulse to between -80 and -90 mV. In the presence of an adenylate cyclase inhibitor, SQ-22536 (100 microM), the intracellular dialysis of ATP also produced a shift in the voltage-dependence of Ih in rat DRG neurons, indicating that the effect of ATP was not caused by cAMP converted by adenylate cyclase. Intracellular dialysis of a nonhydrolysable ATP analog, AMP-PNP or ATP-gamma-S, also produced a positive shift in the voltage-dependence of Ih activation, suggesting that the effect of ATP results from its direct action on the channel protein. These results indicate that cytosolic ATP directly regulates the voltage dependence of Ih activation as an intracellular modulating factor.     

Effects of thyroxine on hyperkalemia and renal cortical Na+, K+ - ATPase activity induced by cyclosporin A

Cyclosporin A (CsA)-induced hyperkalemia is caused by alterations in transepithelial K+ secretion resulting from the inhibition of renal tubular Na+, K+ -ATPase activity. Thyroxine enhances renal cortical Na+, K+ -ATPase activity. This study investigated the effect of thyroxine on CsA-induced hyperkalemia. Sprague-Dawley rats were treated with either CsA, thyroxine, CsA and thyroxine, or olive-oil vehicle. CsA resulted in an increase in BUN and serum K+, along with a decrease in creatinine clearance, fractional excretion of potassium, and renal cortical Na+, K+ -ATPase activity, as compared with oil vehicle administration. Histochemical study showed reduced Na+, K+ -ATPase activity in the proximal tubular epithelial cells of the CsA-treated compared with the oil-treated rats. Histologically, isometric intracytoplasmic vacuolation, disruption of the arrangement and swelling of the mitochondria, and a large number of lysosomes in the tubular epithelium were characteristic of the CsA-treated rats. Co-administration of thyroxine prevented CsA-induced hyperkalemia and reduced creatinine clearance, Na+, K+ -ATPase activity, and severity of the histologic changes in the renal tubular cells when compared with the CsA-treated rats. Thyroxine increased the fractional excretion of potassium via the preservation of Na+, K+ -ATPase activity in the renal tubular cells. Thus, the beneficial effects of thyroxine may be suited to treatment modalities for CsA-induced hyperkalemia.     

血管内皮损伤标志物在肾移植患者中的应用价值

目的 :通过对肾移植患者血浆可溶性血栓调节蛋白 (solublethrombomdulin ,sTM)、可溶性血管内皮细胞蛋白C受体 (endothelialproteinCreceptor ,sEPCR)、血管性血友病因子 (vonWillebrandfactor,vWF)含量水平的动态检测分析 ,探讨其在肾移植及移植排斥反应中的应用价值。方法 :采用双抗体夹心ELISA法检测 4 2例肾移植患者手术前后、排斥反应治疗过程中的sTM、sEPCR、vWF含量。结果 :肾移植术后sTM、sEPCR、vWF含量水平较术前明显增高 (P <0 0 5 )、与正常对照组比较有显著性差异 (P <0 0 1) ;发生移植排斥反应者较无排斥者有明显差异 (P <0 0 5 ) ;肾移植术前、排斥反应有效治疗后与正常对照组比较无显著差异 (P >0 0 5 ) ;肾移植患者sTM、sEPCR、vWF各指标间呈正相关 (r =0 5 95 ,P <0 0 1,r =0 5 81,P <0 0 1,r =0 6 0 6 ,P <0 0 1)。结论 :sTM、sEPCR、vWF均可作为肾移植患者血管内皮的免疫损伤标志物 ,联合动态监测sTM、sEPCR、vWF的含量在观察肾移植手术创伤程度、排斥反应的早期诊断和治疗效果的监测等方面均有重要的临床应用价值。     

髋臼骨关节面形态特征的研究

髋臼骨关节面生物形态特征的研究将有助于通过建立更为精确的髋臼三维模型来分析髋关节的生物力学性能 ,具有重要的临床意义。但在以往的研究中 ,髋臼骨关节面大多被简单认定为一球面。本研究通过三维激光扫描获取髋臼的三维形态数据 -点云数据 ,运用反球工程技术结合优化拟合算法 ,分别用球面和旋转椭球面逐步逼近原始的髋臼骨关节面 ,获得最佳的髋臼骨关节面匹配模型 ,分析比较此二项匹配模型间的匹配误差。对 15例髋骨的统计测量结果表明 ,旋转椭球面的匹配误差显著小于球面的匹配误差 ,其中拟合球半径平均值为 2 4 .37±2 .2 2 mm,拟合旋转椭球面长轴 (髋臼左右方向 )平均值为 2 6 .0 2± 2 .76 mm,短轴 (髋臼前后方向 )平均值为 2 4 .17± 2 .16 mm。该项研究首次对髋臼骨关节面形态作了定量的分析 ,有助于我们对髋臼骨形态的重新认识 ,进而为一系列以其为基础的相关研究及应用提供重要的参考