环氧化酶-2抑制剂塞来昔布对C6胶质瘤细胞增殖和凋亡的影响

目的探讨选择性环氧化酶-2抑制剂塞来昔布对体外培养的C6胶质瘤细胞增殖和凋亡的影响。方法以不同浓度塞来昔布分别作用于C6胶质瘤细胞不同时间段,采用甲基噻唑蓝比色法检测细胞活性,流式细胞仪检测细胞周期和细胞凋亡,并用光镜及透射电镜观察细胞形态学和超微结构的变化。结果经不同浓度塞来昔布作用后,C6胶质瘤细胞活性受到抑制,且呈剂量时间依赖性,实验组与对照组之间以及实验组各亚组之间均有显著性差异（P〈0.05）。细胞周期检测发现实验组S期细胞减少,G2期细胞显著增加,与对照组比较有显著性差异（P〈0.05）;同时,塞来昔布还可诱导C6胶质瘤细胞凋亡,凋亡率在实验组与对照组之间有显著性差异（P〈0.05）。结论选择性环氧化酶-2抑制剂塞来昔布可抑制C6胶质瘤细胞增殖,诱导C6胶质瘤细胞凋亡。     

大黄对重型颅脑损伤后应激性溃疡的预防作用

目的 探讨重型颅脑损伤后应激性溃疡的发生机制及大黄对重型颅脑损伤后应激性溃疡的预防作用.方法 成年雄性SD大鼠40只按随机数字表法分为假手术组、模型组、大黄治疗组和奥美拉唑治疗组,每组10只.后3组大鼠采用改良的Feeney自由落体法制备重型颅脑损伤模型,大黄治疗组和奥美拉唑治疗组大鼠在模型后分别给予大黄粉悬浊液灌胃和奥美拉唑口服.模型后2、6、24 h对各组大鼠行神经损害严重程度评分(NSS);于模型后24 h采用激光血流仪探头检测各组大鼠胃黏膜血流(pu),取胃前壁组织观察并测量黏液层厚度(μm);HE染色观察胃黏膜的病理变化并计算溃疡指数(UI).结果 与假手术组比较,模型组、大黄治疗组和奥美拉唑治疗组大鼠NSS评分较低,差异有统计学意义(P＜0.05);与模型组比较,大黄治疗组和奥美拉唑治疗组大鼠模型后24 h胃黏膜血流和黏液层厚度增加,溃疡指数降低,且大黄治疗组大鼠胃黏膜血流和黏液层厚度均高于奥美拉唑治疗组,差异有统计学意义(P＜0.05);大鼠NSS评分、胃黏膜血流、黏液层厚度与溃疡指数之间均呈现明显的负相关关系(r=-0.429,P=0.002;r=-0.542,P=0.013;r=-0.465,P=0.024).而胃黏膜血流和胃黏膜黏液层厚度之间呈正相关关系(r=0.680,P=0.001).结论 大黄对颅脑损伤后应激性溃疡有预防性作用,且不减少黏液的分泌,但对颅脑损伤大鼠短期内的行为学评分无明显改善.

Abstract：

Objective To explore the mechanism of stress ulcer induced by severe brain injury and prophylactic effect of rhubarb on this stress ulcer in rats. Methods Forty adult male SD rats were equally randomized into sham-operated group, model group, rhubarb treatment group and omeprazole treatment group (n=10). Rat models of craniocerebral injury in the later 3 groups were established according to modifiod Feeney's method; and then, rhubarb in a suspension manner was given by way of ig into the rhubarb treatment group and omeprazole was given orally to rats in the omeprazole treatment group. The neurological severity scale (NSS) was performed 2, 6 and 24 h after the brain injury. The gastric mucosal blood flow (GMBF) was measured by probe test with laser Joppler flowmetry; thickness of the slime layer was measured; pathological changes of mucous mermbrane were observed by HE staining and the ulcer index was calculated 24 h after the brain injury. Results As compared with the sham-operated group, the other 3 groups enjoyed significantly lower scores of NSS (P＜0.05). Thickness of the slime layer and GMBF in the rhubarb and omeprazole treatment groups were obviously increased,and ulcer index in them was obviously decreased as compared with those in the model group (P＜0.05);the levels of thickness of the slime layer and GMBF in the rhubarb treatment group were significantly higher than those in the omeprazole treatment group (P＜0.05). The scores of NSS, thickness of the slime layer and GMBF were negatively correlated to the ulcer index(r=-0.429, P=0.002; r=-0.542, P=0.013;r=-0.465, P=0.024). Positive correlation was noted between thickness of the slime layer and GMBF(r=0.680, P=0.001). Conclusion Prophylactic effect ofrhubarb on severe brain injury induced stress ulcer is noted in rats. But the rhubarb may neither reduce the mucus secretion nor improve the scores of behavioral evaluation in a short term.     

NT4-A1融合基因原核表达载体的构建

目的 构建神经营养素(NT4)与肿瘤血管抑制肽Alphastatin融合基因的原核表达载体pBV220-NT4-A1.方法 采用非对称互补引物(模板)法,依据GenBank提供的Alphastatin基因序列,设计合成并扩增出肿瘤血管抑制肽Alphastatin的cDNA:将其连接到NT4的信号肽和前导序列3'端,组成融合基因NT4-A1,再将该融合基因亚克隆于表达载体pBV220,构建pBV220-NT4-A1.结果 经DNA测序、限制性内切酶酶切等方法 证实Alphastatin成功重组到NT4信号肽和前导序列3'端,并将融合基因亚克隆于pBV220内.结论 成功构建原核表达载体pBV220-NT4-A1,为进一步研究肿瘤基因治疗奠定了基础.     

NT4-Al融合基因原核表达载体的构建

目的构建神经营养素（NT4）与肿瘤血管抑制肽Alphastatin融合基因的原核表达载体pBV220-NT4-Al。方法采用非对称互补引物（模板）法,依据GenBank提供的Alphastatin基因序列,设计合成并扩增出肿瘤血管抑制肽Alphastatin的cDNA;将其连接到NT4的信号肽和前导序列3’端,组成融合基因NT4-Al,再将该融合基因亚克隆于表达载体pBV220,构建pBV220-NT4-Al。结果经DNA测序、限制性内切酶酶切等方法证实Alphastatin成功重组到NT4信号肽和前导序列3’端,并将融合基因亚克隆于pBV220内。结论成功构建原核表达载体pBV220-NT4-Al,为进一步研究肿瘤基因治疗奠定了基础。