肾素血管紧张素拮抗剂对血管紧张素Ⅱ诱导的基质金属蛋白酶高表达的抑制作用

目的 研究血管紧张素转换酶抑制剂卡托普利和血管紧张素Ⅱ(AngⅡ)受体拮抗剂氯沙坦对AngⅡ诱导的血管平滑肌细胞基质金属蛋白酶-1(MMP-1)和MMP-9 mRNA表达的干预作用.方法 体外原代培养雄性Wistar大鼠胸主动脉血管平滑肌细胞,分为对照组、AngⅡ组、卡托普利组、氯沙坦组和卡托普利与氯沙坦联合应用组.收集各组培养终点细胞,以反转录聚合酶链反应(RT-PCR)检测所收集标本中MMP-1和MMP-9 mRNA的表达,观察不同浓度AngⅡ和作用时间对血管平滑肌细胞MMP-1、MMP-9 mRNA表达的影响及卡托普利、氯沙坦的干预作用.结果 MMP-1 mRNA表达随AngⅡ浓度增加及作用时间延长而增加(P＜0.05),AngⅡ浓度为10-6mol/L时最为显著(P＜0.01).一定浓度的卡托普利(5×10-6mol/L)和氯沙坦(5×10-6mmol/L)可抑制AngⅡ的作用(P＜0.05,P＜0.01);AngⅡ为10-7、10-6、10-5、10-4mol/L时,MMP-9 mRNA表达量分别为0.47±0.03、0.86±0.04、0.94±0.14和1.12±0.19,与对照组0.10±0.04比较,差异有统计学意义(P＜0.05或P＜0.01).卡托普利和氯沙坦可抑制AngⅡ对血管平滑肌细胞分泌的MMP-9 mRNA的促进表达作用,以卡托普利和氯沙坦联合应用时抑制作用最强;MMP-9 mRNA的表达随AngⅡ作用时间延长而增加;MMP-9 mRNA较MMP-1表达早.结论 AngⅡ可以诱导血管平滑肌细胞分泌的MMP-1和MMP-9 mRNA高表达,且存在剂量和时间依赖性.卡托普利、氯沙坦可抑制AngⅡ诱导的血管平滑肌细胞MMP-1和MMP-9 mRNA高表达;卡托普利和氯沙坦联合应用时抑制作用最强;这种抑制作用与作用时间相关.

Abstract：

Objective To investigate the effect of angiotensin converting enzyme inhibitor (ACEI) captopril and angiotensin Ⅱ receptor antagonist losartan on the mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-9 in vascular smooth muscle cells induced by angiotensin Ⅱ (Ang Ⅱ ).Methods Male Wistar rats' thoracic aortic vascular smooth muscle cells were cultured in vitro.The cultured cells were divided in to control group,Ang Ⅱ group,captopril group,losartan group,and captopril plus losartan group.Cells in all groups were collected at the culture end-point.MMP-1 and MMP-9 mRNA expressions were detected by RT-PCR method in the collected specimens,and the effects of Ang Ⅱ on MMP-1 and MMP-9 mRNA expression and the intervention effects of captopril and losartan were observed in different Ang Ⅱ concentrations and different action times to vascular smooth muscle cells.Results ( 1 ) MMP-1 mRNA expression gradually increased along with the increments of Ang Ⅱ concentration and the action time (P＜0.05),and the most significant concentration was 10-6 mol/L (P＜0.01).(2)Captopril (5 × 10-6 mol/L) and losartan (5 × 10-6mol/L) inhibited the action of AngⅡ (P＜0.05,P＜0.01).MMP-9 mRNA expression was 0.47±0.03 ,0.86 ± 0.04,0.94±0.14 and 1.12±0.19 vs.0.10±0.04 (P＜0.05,P＜0.01) respectively when Ang Ⅱ concentration was 10-7 ,10-6 ,10-5 and 10-4 mol/L respectively.Captopril (5 × 10-6mol/L) and losartan (5 × 10-6 mol/I) significantly inhibited the MMP-9 mRNA expression which was stimulated by Ang Ⅱ (P＜0.05,P＜0.01),especially in captopril plus losartan group.The MMP-9 mRNA expression increased with the prolonging of stimulating time of Ang Ⅱ,MMP-9 mRNA expression was earlier than that of MMP-1.Conclusions AngⅡ increases the expression of MMP-1 and MMP-9 of vascular smooth muscle cells in a dose-and time-dependence manner.Captopril and losartan inhibit the MMP-1 and MMP-9 mRNA expression of vascular smooth muscle cells induced by Ang Ⅱ ,and the inhibition is the strongest when losartan was combined with captopril.The inhibitive effects is positively correlated to action time.     

Inhibition effect of captopril and losartan on expression of matrix metalloproteinase-1,-9 induced by angiotensin Ⅱ in rat vascular smooth muscle cells

Objective To investigate the effect of angiotensin converting enzyme inhibitor (ACEI) captopril and angiotensin Ⅱ receptor antagonist losartan on the mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-9 in vascular smooth muscle cells induced by angiotensin Ⅱ (Ang Ⅱ ).Methods Male Wistar rats' thoracic aortic vascular smooth muscle cells were cultured in vitro.The cultured cells were divided in to control group,Ang Ⅱ group,captopril group,losartan group,and captopril plus losartan group.Cells in all groups were collected at the culture end-point.MMP-1 and MMP-9 mRNA expressions were detected by RT-PCR method in the collected specimens,and the effects of Ang Ⅱ on MMP-1 and MMP-9 mRNA expression and the intervention effects of captopril and losartan were observed in different Ang Ⅱ concentrations and different action times to vascular smooth muscle cells.Results ( 1 ) MMP-1 mRNA expression gradually increased along with the increments of Ang Ⅱ concentration and the action time (P＜0.05),and the most significant concentration was 10-6 mol/L (P＜0.01).(2)Captopril (5 × 10-6 mol/L) and losartan (5 × 10-6mol/L) inhibited the action of AngⅡ (P＜0.05,P＜0.01).MMP-9 mRNA expression was 0.47±0.03 ,0.86 ± 0.04,0.94±0.14 and 1.12±0.19 vs.0.10±0.04 (P＜0.05,P＜0.01) respectively when Ang Ⅱ concentration was 10-7 ,10-6 ,10-5 and 10-4 mol/L respectively.Captopril (5 × 10-6mol/L) and losartan (5 × 10-6 mol/I) significantly inhibited the MMP-9 mRNA expression which was stimulated by Ang Ⅱ (P＜0.05,P＜0.01),especially in captopril plus losartan group.The MMP-9 mRNA expression increased with the prolonging of stimulating time of Ang Ⅱ,MMP-9 mRNA expression was earlier than that of MMP-1.Conclusions AngⅡ increases the expression of MMP-1 and MMP-9 of vascular smooth muscle cells in a dose-and time-dependence manner.Captopril and losartan inhibit the MMP-1 and MMP-9 mRNA expression of vascular smooth muscle cells induced by Ang Ⅱ ,and the inhibition is the strongest when losartan was combined with captopril.The inhibitive effects is positively correlated to action time.     

Ⅱ、Ⅲ期直肠癌根治术后不同氟脲嘧啶联合放化方案效果比较

目的比较三种不同氟脲嘧啶联合放化方案在Ⅱ、Ⅲ期直肠癌根治术后辅助治疗的疗效及不良反应。方法对2005年11月至2008年2月对我科收治的43例Ⅱ、Ⅲ期直肠癌随机分组：AmA放疗第1～5,29～33天静脉推注5-氟尿嘧啶（5-FU）250mg／（m^2·d）,5-FU之前使用亚叶酸钙（OF）200mg／（m^2·d）增敏,同时盆腔放疗;ArmB放疗第1～5,29～33天持续静脉泵注5．氟尿嘧啶（5-FU）700mg／（m^2·d）,同时盆腔放疗;ArmC放疗第1～14,22～35天口服卡培他滨1600mg／（m^2·d）,同时盆腔放疗50Gy。序贯FOLFOX4方案全身化疗6个周期。结果ArrnAIArmB和ArmC三组两年0s分别为86％、89％和92％（P〉0．05）,两年DFS分别为64％、69％和77％（P〉0．05）。放化各组3～4级急性毒副反应不同,ArmA主要为骨髓抑制,ArmB主要为3～4级放射性直肠炎或手足综合征,ArmC副反应较轻。结论三种不同氟脲嘧啶联合放化治疗0s差异无统计学意义,希罗达联合放疗组具有延长D鹏的趋势。三种不同氟脲嘧啶联合放化均安全可耐受,希罗达联合放疗组毒副反应较轻。     

应该对PVS病人进行生命维持吗？——庄子思想的启示

庄子的生命观消解了生命神圣论，主张以安时处顺、恬淡自然的态度提高生命质量；提醒人们应该清醒意识到生命的极限，反对对生命进行“遁天倍情”的妄为之举。现从庄子的生命观出发，将使解决这一伦理难题得到启示。     

毛细管气相色谱法测定蔬菜中有机氯和拟除虫菊酯类农药残留

目的:应用毛细管柱气相色谱法测定蔬菜中三氯杀螨醇和4种拟除虫菊酯类农药残留量。方法:样品经乙腈提取和氯化钠盐析分层,再经硅镁柱净化,氮气流吹干后正己烷溶解定容,采用DB-1701毛细管柱分离,ECD检测,外标法定量。结果:平均加标回收率为80%～110.1%,相对标准偏差为1.3%～4.6%,检出限为0.12μg/kg～2.6μg/kg。结论:该方法准确、灵敏、可靠,适用于批量蔬菜样品的分析。     

肝癌细胞来源的exosomes提取方法的改进及电镜观察

目的：分离提纯两种肝癌细胞HepG2细胞和Hep3B细胞来源的exosomes，并在透射电镜下观察exosomes的形态特征，为肿瘤免疫治疗寻找肿瘤新抗原提供实验材料。方法：采用离心超滤联合蔗糖密度梯度离心等方法分离出HepG2和Hep3B细胞释放exosomes，经透射电镜观察其超微结构。结果：HepG2和Hep3B细胞分泌释放的exosomes为直径50-100nm的膜性微囊结构，圆形或椭圆形，腔内为低电子密度成分。结论：离心超滤和蔗糖密度梯度离心法分离提纯exosomes方法是较为简便高效，对肿瘤细胞培养上清提纯exosomes而言具有可行性。     

乳源致病性金黄葡萄球菌原生质体膜表面蛋白nEBPS的亚细胞定位研究

目的克隆牛乳源致病性金黄葡萄球菌nEBPS基因,经工程菌表达得到其编码抗原纯化蛋白,制备多克隆抗体,采用间接荧光抗体法鉴定nEBPS蛋白。方法设计1对克隆引物,PCR扩增牛乳腺炎金黄葡萄球菌内蒙古分离株EBPS基因序列,构建重组表达质粒,转化工程菌,表达膜表面亚单位蛋白nEBPS,纯化后免疫家兔制备抗nEBPS蛋白抗体,采用间接荧光抗体法对乳源致病性金黄葡萄球菌原生质体膜表面蛋白nEBPS进行亚细胞定位。结果在含25μg/ml Amp、0.015g/ml NaCl的LB液体培养基中成功培养制备了金黄葡萄球菌原生质体。对原生质体细胞(表面)和利用0.02g/ml NaCl高渗溶液裂解原生质体细胞离心获得的细胞膜残片进行间接荧光抗体试验,均出现特异荧光。结论间接荧光抗体试验鉴定nEBPS蛋白为膜表面蛋白,可作为乳及乳制品金黄葡萄球菌检测和制备免疫制剂的靶抗原。     

健脾消痞汤治疗功能性消化不良的胃动力学超声改变及其机制探讨

[目的]观察中药治疗对功能性消化不良(FD)患者的胃动力学变化,并探讨其作用机制。[方法]将40例FD患者用健脾消痞汤加减治疗1个疗程,观察对比治疗前后的胃动力学改变以及胃动素(MOT)、生长抑素(SS)的分泌情况。[结果]治疗后总有效率达97.5%;超声胃动力学指标:胃窦收缩幅度(△V)为(518.45±160.40)mm2、胃窦收缩频率(F)为(2.80±0.70)次/min、胃窦运动指数(MI)为(1 450.66±538.75)、胃窦半排空时间(T1/2)为(29.93±0.14)min、胃窦排空时间(T)为(46.35±13.82)min。△V较治疗前提高(P0.05),F较治疗前增加(P0.05)、MI较治疗前显著改善(P0.01),T1/2、T均较治疗前明显缩短(P0.01);患者空腹MOT较治疗前明显增高(P0.01)、SS较治疗前明显减低(P0.01)。[结论]健脾消痞汤治疗FD疗效满意,能够改善胃的排空功能,其作用机制可能是通过神经-体液调节各种胃肠激素的分泌所致。     

塞来昔布对乳腺癌阿霉素耐药细胞株MCF-7/ADM的增殖抑制作用

目的： 通过研究选择性环氯化酶2(COX-2)抑制剂塞来昔布联合阿霉素(ADM)对乳腺癌MCF-7/ADM细胞株生长的作用,探讨塞来昔布的抗肿瘤作用。方法：MCF-7/ADM细胞加入不同浓度等倍稀释的ADM（0.05、0.10、0.20、0.40、0.80和1.60 mg/L）为对照组,MCF-7/ADM细胞加入无细胞的完全培养基为阴性对照组,在对照组的基础上联合应用10、20 μmol/L塞来昔布为实验组,各组细胞于24、48、72 h后采用CCK-8检测细胞生长抑制率。MCF-7/ADM细胞单加0.05 mg?L-1ADM及联合塞来昔布(10、20 μmol/L)处理后3 h收集细胞,采用流式细胞术检测细胞内ADM水平。结果：不同浓度ADM单药及联合塞来昔布（10和20 μmol/L）　作用于MCF-7/ADM 24、48和72 h后细胞生长受到抑制,实验组不同时间细胞生长抑制率均高于对照组(P<0.05),且20 μmol/L塞来昔布实验组细胞生长抑制率高于10 μmol?L-1塞来昔布实验组,差异具有统计学意义（P<0.05）。对照组24、48和72 h的细胞未见明显变化,但实验组细胞作用48和72 h后出现细胞改变及死亡,呈塞来昔布剂量依赖性。与对照组比较,塞来昔布实验组（10和20 μmol/L）细胞内ADM浓度升高（P<0.05）;20 μmol/L塞来昔布实验组细胞内ADM浓度高于 10 μmol/L塞来昔布实验组(P<0.05)。结论：选择性COX-2抑制剂塞来昔布联合ADM可有效逆转乳腺癌ADM细胞株耐药。
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EFFECTS OF VARYING INSPIRATORY FLOW WAVEFORM AND TIME IN INTERMITTENT POSITIVE PRESSURE VENTILATION: PULMONARY OEDEMA

Greyhound dogs were given oleic acid 1. v to induce controlledpulmonary oedema. These animals were then studied using intermittentpositive pressure ventilation (IPPV) with four different inspiratoryflow waveforms, each at three different inspiratory times, ina fixed respiratory cycle of 4s at a constant tidal volume.Although there were statistically significant differences inairway and oesophageal pressures between the different waveformsand times, there was little variation in the other physiologicalparameters studied except for arterial carbon dioxide tension(Pa_CO2) which showed statistically significant improvement withreversed ramp and sine wave inputs and at the longer inspiratorytime of 2.2s Venous admixture was also less with the longerinspiratory time of 2.2 s.