糖化终末产物对内皮细胞氧化应激的影响

目的 研究糖化终末产物(AGEs)对猪髂动脉内皮细胞(PIEC)氧化应激的影响.方法 不同浓度AGEs干预PIEC一定时间后,MTT比色法检测细胞活力变化.运用活性氧(ROS)捕获剂双氢-乙酰乙酸二氯荧光黄(DCFH-DA)孵育细胞,通过流式细胞仪检测细胞内二氯荧光黄(DCF)的荧光强度而测得细胞内ROS水平.结果 随着AGEs浓度的增加和作用时间的延长,PIEC活力明显下降(P＜0.05);细胞活力下降与作用浓度及时间呈显著正相关(r=0.927,P＜0.01).经DCFH-DA孵育后流式细胞仪检测显示,AGEs处理组细胞内DCF平均荧光强度较空白对照组明显升高.结论 AGEs抑制内皮细胞活力,使细胞内ROS生成明显增加,直接诱导内皮细胞的氧化应激.     

高糖对人腹膜间皮细胞氧化应激的影响

目的研究不同浓度葡萄糖对人腹膜间皮细胞(HPMC)氧化应激的影响。方法HPMC体外培养并行免疫组化鉴定,取第2代细胞用于实验。将不同浓度葡萄糖干预HPMC一定时间后,MTT比色法检测细胞活力;采用活性氧捕获剂双氢-乙酰乙酸二氯荧光黄(DCFH-DA)孵育细胞,通过流式细胞仪检测细胞内的荧光强度而测得细胞内活性氧(ROS)水平。分析不同浓度葡萄糖及不同作用时间对HPMC细胞活力及ROS水平的影响。结果高糖明显抑制HPMC细胞活力(P<0.05),随着葡萄糖浓度增加及作用时间的延长,抑制率呈上升趋势。高糖干预HPMC后,细胞内的平均荧光强度明显升高。结论高糖导致细胞发生氧化应激,其对细胞的损伤作用可能与氧自由基生成过多及ROS蓄积有关。     

高糖对人腹膜间皮细胞氧化应激的影响

目的 研究不同浓度葡萄糖对人腹膜间皮细胞(HPMC)氧化应激的影响.方法 HPMC体外培养并行免疫组化鉴定,取第2代细胞用于实验.将不同浓度葡萄糖干预HPMC一定时间后,MTT比色法检测细胞活力;采用活性氧捕获剂双氢-乙酰乙酸二氯荧光黄(DCFH-DA)孵育细胞,通过流式细胞仪检测细胞内的荧光强度而测得细胞内活性氧(ROS)水平.分析不同浓度葡萄糖及不同作用时间对HPMC细胞活力及ROS水平的影响.结果 高糖明显抑制HPMC细胞活力(P＜0.05),随着葡萄糖浓度增加及作用时间的延长,抑制率呈上升趋势.高糖干预HPMC后,细胞内的平均荧光强度明显升高.结论 高糖导致细胞发生氧化应激,其对细胞的损伤作用可能与氧自由基生成过多及ROS蓄积有关.     

格列吡嗪和格列奇特对MIN6细胞的影响

目的:研究不同磺脲类药物对胰岛素分泌细胞的影响及机制。方法:不同浓度格列吡嗪和格列奇特干预MIN6细胞一定时间后,MTT法检测细胞活力变化,运用ROS捕获剂双氢溴乙啶(HE)、双氢-乙酰乙酸二氯荧光黄(DCFH-DA)和双氢罗丹明123(DHR123)孵育细胞,用荧光显微镜观察细胞内荧光、流式细胞仪检测MIN6细胞的平均荧光强度(MFI)而测得细胞内ROS水平。用Annexin-vFITC和PI双标记细胞凋亡检测法通过流式细胞仪测定细胞凋亡情况。结果:格列吡嗪干预MIN6细胞后,细胞活力明显被抑制(P<0.05);细胞内的MFI明显增加;细胞凋亡率明显增加;而且,细胞活力下降、MFI和细胞凋亡率与药物作用浓度和时间呈正相关。而格列奇特干预MIN6细胞后,细胞活力无明显抑制(P>0.05);细胞内的MFI无明显增加;细胞凋亡率亦无明显增加。结论:格列吡嗪可诱导MIN6细胞发生氧化应激,诱导MIN6细胞凋亡,说明临床上磺脲类药物治疗糖尿病出现失效可能与磺脲类药物诱发胰岛细胞衰竭相关,然而,格列奇特不诱导MIN6细胞损害,说明并非所有磺脲类药物对β细胞都有损害作用。     

替米沙坦对MIN6细胞的影响

目的:研究血管经张素Ⅱ受体拮抗剂(ARB)药物对胰岛素分泌细胞的影响及机制。方法:不同浓度替米沙坦及替米沙坦和甲苯磺丁脲共同干预MIN6细胞一定时间后,MTT法检测细胞活力变化,运用ROS(细胞内活性氧)捕获剂双氢溴乙啶(HE)、双氢-乙酰乙酸二氯荧光黄(DCFH-DA)和双氢罗丹明123(DHR123)孵育细胞,用荧光显微镜观察细胞内荧光、流式细胞仪检测MIN6细胞的MFI(平均荧光强度)而测得细胞内ROS水平。用Annexin-vFITC和PI双标记细胞凋亡检测法通过流式细胞仪测定细胞凋亡情况。结果:替米沙坦单独干预MIN6细胞后,细胞活力无明显抑制;细胞内的MFI无明显增加;细胞凋亡率亦无明显增加;而替米沙坦和甲苯磺丁脲共同孵育MIN6细胞后,细胞活力相对甲苯磺丁脲单独干预时增强(P>0.05);细胞内的MFI明显减少;细胞凋亡率亦明显减少(前期实验已经做过甲苯磺丁脲对MIN6细胞影响)。结论:替米沙坦对甲苯磺丁脲诱导的MIN6细胞氧化应激和凋亡有保护作用。说明ARB类药物对胰岛β细胞功能有保护作用,延缓β细胞衰竭。     

细胞内活性氧在晚期糖基化终产物促肾小球系膜细胞合成纤维连接蛋白中的作用

目的 观察晚期糖基化终末产物(AGEs)对大鼠肾小球系膜细胞合成和分泌纤维连接蛋白的影响及细胞内活性氧(ROS)在其中的作用.方法 在体外制备AGEs,分别用不同浓度的(0,100,500,1000 μg/mL)AGEs及抗氧化剂N-酰-L-半胱氨酸(NAC 30 mmol/L)作用于肾小球系膜细胞,以氧化敏感的荧光染料2、7-二氢二氯荧光素(DCFH)染色,流式细胞仪测定细胞内ROS强度;再用RT-PCR和ELISA方法测定系膜细胞中纤维连接蛋白的表达.结果 AGEs能使细胞内ROS水平明显升高(P<0. 05),呈现浓度依赖效应;AGEs同时以时效和量效方式促进肾小球系膜细胞中纤维连接蛋白基因和蛋白的表达(P<0.05);而NAC能够明显抑制AGEs所致的细胞内ROS升高,同时抑制肾小球系膜细胞中纤维连接蛋白的合成和分泌(P<0. 05).结论 AGEs可能部分通过诱导细胞内ROS,促进肾小球系膜细胞表达分泌纤维连接蛋白,从而引起肾小球系膜基质增生,最终导致肾小球纤维化,而抗氧化治疗可能有助于阻止糖尿病肾病的氧化应激损害和肾小球纤维化的发展进程.     

高浓度葡萄糖在内皮细胞氧化应激中的作用

目的 探讨高糖对猪髂动脉内皮细胞(PIECs)氧化应激的影响.方法 不同浓度高糖干预PIECs一定时间后,应用活性氧(ROS)捕获剂dihydroethidium(DHE)、双氢罗丹明123(DHR123)孵育细胞,通过荧光显微镜观察细胞内DHE染色,流式细胞仪检测细胞内罗丹明123(rh123)的平均荧光强度(MFI)而测得细胞内ROS水平;以二甲基吖啶硝酸盐为化学发光试剂,通过化学发光仪检测加入NADPH后发光值的变化,以反映高糖对PIECs中NADPH氧化酶(NOX)活性的影响.设立正常组和甘露醇组作为对照.结果 随着高浓度葡萄糖作用浓度和时间的增加,荧光显微镜下高糖组细胞内DHE荧光增强;流式细胞仪检测显示,高糖组细胞内rh123的MFI较对照组明显升高;化学发光仪检测表明,高糖组细胞NOX活性升高显著.结论 高糖导致细胞发生氧化应激,其对细胞的损伤作用可能与氧自由基生成过多、ROS蓄积有关,该效应可能由NOX介导.     

高浓度葡萄糖在内皮细胞氧化应激中的作用

目的探讨高糖对猪髂动脉内皮细胞(PIECs)氧化应激的影响。方法不同浓度高糖干预PIECs一定时间后,应用活性氧(ROS)捕获剂dihydroethidium(DHE)、双氢罗丹明123(DHR123)孵育细胞,通过荧光显微镜观察细胞内DHE染色,流式细胞仪检测细胞内罗丹明123(rh123)的平均荧光强度(MFI)而测得细胞内ROS水平;以二甲基吖啶硝酸盐为化学发光试剂,通过化学发光仪检测加入NADPH后发光值的变化,以反映高糖对PIECs中NADPH氧化酶(NOX)活性的影响。设立正常组和甘露醇组作为对照。结果随着高浓度葡萄糖作用浓度和时间的增加,荧光显微镜下高糖组细胞内DHE荧光增强;流式细胞仪检测显示,高糖组细胞内rh123的MFI较对照组明显升高;化学发光仪检测表明,高糖组细胞NOX活性升高显著。结论高糖导致细胞发生氧化应激,其对细胞的损伤作用可能与氧自由基生成过多、ROS蓄积有关,该效应可能由NOX介导。     

甲基乙二醛诱导大鼠海马神经元氧化应激损伤的实验研究

目的:探讨甲基乙二醛对大鼠海马神经元的损伤作用及其可能机制。方法:取新生24 h的SD大鼠分离并培养其海马神经元,第7天予甲基乙二醛(MGO)或(和)N-乙酰半胱氨酸(NAC)干预24 h,四甲基偶氮唑蓝(MTT)法检测MGO或(和)NAC对海马神经元活力的影响,并以氧化敏感的2,7-二氢二氯荧光素(DCFH-DA)染色,荧光显微镜观察及流式细胞仪测定细胞内活性氧(ROS)强度。结果:MTT法检测结果显示,随MGO浓度增加,海马神经元存活率逐渐降低(均P<0.05),而NAC能显著增加海马神经元存活率,且10 mmol.L-1组存活率最高(P<0.01);100μmol.L-1MGO能使细胞内ROS水平明显增加(P<0.01),而同时加入10 mmol.L-1NAC能够抑制MGO导致的细胞内ROS增高。结论:MGO可能部分通过诱导细胞内ROS产生,导致神经元氧化应激损伤,而NAC可能通过降低ROS的水平而对神经元起保护作用。     

参附注射液对氧化应激血管内皮损伤及相关信号转导通路的影响

目的 研究参附注射液对氧化应激血管内皮损伤的作用及相关信号转导通路的影响.方法 培养血管内皮细胞ECV304,MTT法筛选H2 O2造模和参附注射液的干预浓度;DCFH-DA荧光探针检测细胞内活性氧(ROS)含量;Hoechst染色和流式细胞术检测评估细胞凋亡;Western blot法检测Cleaved Caspas...     

Value of D-Dimers in patients with acute aortic dissection

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.     

Research of combined liver-kidney transplantation model in rats

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

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.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

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.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

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.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

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.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.