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1.
观察反义寡脱氧核苷酸对脯氨酰4-羟化酶两种亚基mRNA表达的影响。方法采用定量RT-PCR技术检测培养的人皮肤成纤维细胞中P4HmRNA的相对含量。含量:不同浓度(0.1,0.2,05,1.0μmol/L)反义ODN处理组,人皮肤成纤维细胞中P4HmRNA表达水平均显著低于对照组及错配ODN处理组,其中0.1、0.2μmol/L反义ODNA作用20h时,抑制效应达到最大,50-70h后,逐渐恢复到  相似文献   

2.
反义寡脱氧核苷酸对人肝癌裸鼠模型生长及转移的影响   总被引:3,自引:0,他引:3  
目的 观察反义H-ras寡脱氧核苷酸(ODN)对高表达蛋白p21H- ras的人肝癌高转移裸鼠模型LCI-D20细胞生长、凋亡及转移潜能的影响。方法原代培养LCI-D20细胞,经10μmol/L浓度反义ODN处理后,以1.5×10细胞数分别接种于14只裸鼠皮下缝制皮囊内:6只接种反义H-rasODN处理细胞,4只接种H-ras非特异性反义ODN处理细胞,余4只接种未作处理的对照细胞。结果反义H-ras对LCI-D20细胞增殖具有显著抑制作用(t=31.529,P<0.01),反义ODN处理细胞,S期比率(36.0±1.4)显著低于对照组(58.5±0.9,t=13.519,P<0.01),而G1/G0期比率(56.7%±1.1%)高于对照组(37.4±0.7,t=14.802,P<0.01);反义H-rasODN处理细胞的凋亡发生率(34.0%±4.5%)显著高于对照组(2.5%±1.2%,t=13.434,P<0.01);反义H-ras处理对LCI-D20细胞在接种动物体内的生长具有显著延迟抑制作用(t=3.509,t=3.452,P<0.01);接种动物的肺转移率,反义H-rasODN处理组显著低于对照组。  相似文献   

3.
反义寡核苷酸体外抑制丙型肝炎病毒的研究   总被引:5,自引:0,他引:5  
刘勇  陈智 《中华医学杂志》1997,77(8):567-570
研究反义寡核苷酸体外抑制丙型肝炎病毒(HCV)的方法。方法以重组丙型肝炎病毒(pCD-HCV)转染的H9细胞为对象,通过半定量逆转录聚合酶链反应和dotELISA检测HCVmRNA和抗原表达的变化,观察与HCV结构区(包括AUG)互补和同源以及随机的硫代磷酸化寡核苷酸(PS-ASON、PS-ODN、rPS-ODN)对HCV的抑制作用。结果PS-ASON和PS-ODN可有效地进入靶细胞并在体外与靶基因杂交结合,终浓度为10μmol/L的PS-ODN、rPS-ODN对HCV均无抑制作用,但PS-ASON可明显降低HCVmRNA和抗原表达水平,具有药物浓度和时间依赖效应,脂质体修饰可增强PS-ASON的反义抑制作用,磷酸钙修饰却无此作用。结论与HCV互补的PS-ASON是HCV的反义寡核苷酸,在翻译水平上具有反义抑制作用。  相似文献   

4.
目的:探讨脓毒血症时心血管系统血红素加氧酶(heme oxygenase,HO)的变化及作用。方法:盲肠结扎穿孔法复制大鼠腹膜炎脓毒血症模型,术后9 h 和18 h 测血流动力学指标及血浆乳酸脱氢酶(lactate dehydrogenase,LDH),取心、主动脉测HO 活性和丙二醛(malondialdehyde,MDA) 含量,Northern Blot 测HO1mRNA 表达。结果:早期组血流动力学指标及血浆LDH 无明显变化,心脏与主动脉MDA 含量分别增加60.3 % 和96 .5 % (P< 0 .05) ,心脏HO 活性比对照组升高72 .1% (P< 0.01) ,主动脉HO 活性无明显变化。晚期组血压降低、心功能下降,血浆LDH与组织MDA含量升高,心脏HO 活性仍维持高水平,主动脉HO 活性比早期组增加261 % (P< 0.01)。心脏HO1 m RNA表达与HO 活性变化趋势相同;早、晚期主动脉HO1mRNA表达分别增加49 .2% (P< 0.05) 和143%( P<0 .01) 。结论:脓毒血症时心脏与主动脉HO 活性及HO1 mRNA 表达增加,但变化趋势不同;HO可能与早期机体的抗损伤机制和晚期低血压的形成有  相似文献   

5.
目的 拟建立逆转录-聚合酶链式反应(RT-PCR)半定量法,探讨纤维蛋白原(Fg)、极低密度脂蛋白(VLDL)对人脐静脉内皮细胞(HUVECs)组织型纤溶酶原激活物(t-PA)、I型纤溶酶原灭活剂(PAI-l)m RNA 表达的影响。 方法 (l)用异硫氰酸胍一步法(TRIzol试剂)提取细胞总RNA,用单管RT-PCR 法、三对引物分别逆转录、扩增t-PA,PAI-1 及内参照三磷酸甘油醛脱氢酶(GAPDH)的特异性片段,扩增产物经2.0% 琼脂糖凝胶电泳分离后,用凝胶图像分析仪照相、计算各条带分子量并扫描其光密度值(integrated density value,IDV),t-PA、PAI-1 m RNA的IDV 用GAPDH m RNA 的IDV 进行校正,其相对表达量用各实验组与对照组比较的倍数表示。(2)以ELISA 法测定内皮细胞(EC)培养上清液(CM)中t-PA,PAI-1总抗原的含量。(3)EC与不同浓度Fg(2,20,200,2000 μg/m l)或VLDL(5,10,20,30,40 μg/m l)共同孵育12 h 或24 h 后,观察ECt-PA 和PAI-1 m RNA 水平及CM 中抗原含量的变化。(4  相似文献   

6.
目的:检测慢性丙型肝炎患者外周血T细胞亚型和NK活性,并与正常人比较,以观察HCV感染对患者细胞免疫功能的影响。方法:采用间接免疫荧光-流式细胞分析法检测外周血T细胞CD3、CD4、CD8表达,采用LDH释放法测定NK活性。结果:慢性丙型肝炎患者外周血CD4、CD4/CD8比值、NK活性明显降低,与正常人比较差异有显著性(P值分别<0.01,<0.01和<0.05),而CD8升高(P值<0.05);HCV-RNA阳性组与HCV-RNA阴性组比较,CD4/CD8比值、NK活性进一步下降,差异有显著性(P值分别<0.05)。结论:慢性丙型肝炎患者存在细胞免疫功能紊乱,提高其细胞免疫功能可能有利于患者的愈后。  相似文献   

7.
应用细胞培养、核酸斑点杂交、~3H-胸腺嘧啶核苷(~3H-TdR)掺入、免疫细胞化学、图像分析等技术观察c-myc反义寡核苷酸(ODNs)对低氧内皮细胞条件培养液(HECCM)诱导的大鼠肺血管周细胞c-myc mRNA表达、增殖细胞核抗原(PCNA)表达及~3H-TdR掺入量的影响。结果表明,HECCM显著促进周细胞 c-myc mRNA表达、PCNA表达(P<0.01)及~3H-TdR掺入量增加(P<0.01),反义ODNs显著阻抑 HECCM诱导的周细胞c-myc mRNA表达、PCNA表达(P<0.01)及~3H-TdR掺入量增加(P<0.05),而同义ODNs无上述作用(P>0.05)。结果提示,HECCM可通过上调c-myc基因表达促进肺血管周细胞增殖,反义ODNs通过下凋c-myc基因表达,抑制HECCM诱导的肺血管周细胞增殖。  相似文献   

8.
原癌基因c-myc参与调控细胞的增殖、分化和凋亡过程,在白血病细胞株HL-60 中有很高的表达。c-myc通过扩增活化或基因重排而异常激活,在白血病的发生和发展中可能起着重要的作用。 目的 通过脂质体介导c-myc反义核酸转染HL-60 细胞,观察其抑制细胞生长增殖、促进分化、诱导凋亡,降低c-mycm RNA 和蛋白表达水平,探讨反义核酸的抗肿瘤作用机制。 方法 HL-60 细胞在37℃、5%CO2 条件下,培养于10% 小牛血清1640 培养液中,细胞接种密度为4×107/L,反义核酸终浓度为1μm ol/L,反义核酸与脂质体比例为1 μm ol(6.5 μg)∶10 μg,无血清转染6 h。实验同时设置空白组、无义组、反义组、脂质体组、脂质体无义组等对照组。连续培养4 天。通过锥虫蓝拒染实验测定细胞的生长能力;NBT还原实验和瑞特-姬姆萨染色测定细胞的分化能力;免疫细胞化学染色检测c-myc蛋白表达;RT-PCR法检测c-myc m RNA 水平;吖啶橙/溴化乙锭染色观察凋亡细胞形态;流式细胞仪检测凋亡率;DNA抽提及电泳观察凋亡细胞DNA降解片段的形成。 结果 脂质体转染c-myc反义核酸的作用:(1)抑制HL-60  相似文献   

9.
目的:了解慢性丙型肝炎患者(2',5')寡腺苷酸合成酶(2-5AS)活性在干扰素治疗过程中的动态变化及意义。方法:PEI-CELLULOSE薄板层析法。结果:(1)慢性丙型肝炎患者2-5AS基础水平比正常人显著升高(P<0.01);(2)IFNα治疗结束后HCVRNA持续阳性的病人2-5AS基础水平比HCVRNA转阴的病人要高(P<0.05);(3)IFNα治疗效果较好,HCVRNA转阴的病人2-5AS水平在用药后24h比HCVRNA未转阴的病人上升得更高(P<0.01);(4)用药后1个月2~5AS水平与IFNα治疗效果无显著相关性(P>0.05)。结论:2-5AS可以作为评价干扰素治疗效果的1个机体反应指标。  相似文献   

10.
目的:观察一氧化氮(NO)对大鼠小肠缺血/再灌注(I/R)的作用及内皮素的改变。方法:复制内脏血管阻塞(SAO)性休克模型,用左旋精氨酸(L-Arg)及硝基精氨酸甲脂(L-NAME)处理动物模型后分别测定血浆中ETs、MDA、组织蛋白酶D(CD)、NO-2/NO-3含量。结果:L-Arg减缓了大鼠I/R血压下降(P<0.01),降低了血浆MPO、LDH、CD、MDA的含量(P<0.01)及肠组织中伊文思蓝(EB)的含量(P<0.05);L-NAME与L-Arg相反。MPO与EB正相关(P<0.01),NO-2/NO-3与ETs负相关(P<0.05)。结论:NO对小肠I/R损伤有保护作用,ETs参与SAO休克过程。  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

15.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

16.
Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.  相似文献   

17.
Objective:To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas(SPTP). Methods:Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin(HE) staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results:Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin(9/9 cases), AAT(9/9 cases), NSE(9/9 cases), ACT(7/9 cases), CK20(2/9 cases), CgA(1/9 cases), S-100(3/gcases), PR(4/gcases), Syn(3/9 cases) and CD56(5/9cases), negative in CEA and ER. Conclusion:SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.  相似文献   

18.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

19.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

20.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

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