三氧化二砷抑制U266细胞增殖、诱导凋亡及与血管内皮生长因子表达的关系

目的 探讨三氧化二砷(As2O3)在体外对骨髓瘤细胞系U266增殖、凋亡的影响及与血管内皮生长因子(VEGF)表达的关系.方法 四唑盐比色法(MTT)观察As2O3对U266细胞增殖的影响;缺口末端标记法(TUNEL)分析As2O3对U266细胞凋亡的影响;逆转录多聚酶链反应(RT-PCR)检测2 μmol/L AS2O3不同处理时间后U266细胞VEGF mRNA表达的变化;酶联免疫吸附试验(ELISA)检测As2O3不同浓度不同处理时间对U266细胞VEGF蛋白表达的变化. 结果 (1)As2 Q3明显抑制U266细胞增殖,半数抑制浓度(IC50)为2 μmol/L;(2)As2O3能诱导U266细胞凋亡,呈剂量依赖性;(3)As2O3(2,5,10μmol/L)分别处理72 h后细胞培养上清中VEGF量呈剂量依赖性减少;(4)考虑细胞增殖对VEGF分泌的影响,2 μmol/L As2O3处理组的上清VEGF量与对照组的差别无统计学意义,RT-PCR显示2μmol/L As2O3处理的U266细胞个体VEGF mRNA的表达无变化.结论 As2O3可以诱导骨髓瘤细胞凋亡,抑制其增殖.As2O3可以通过抑制细胞增殖、诱导凋亡,减少肿瘤负荷,减少VEGF总量,但是不改变细胞个体VEGF的表达.     

三氧化二砷联合抗坏血酸对肝癌细胞增殖和凋亡的影响

目的:研究三氧化二砷(As2O3)联合抗坏血酸(AA)对肝癌HepG2细胞增殖和凋亡的影响,并探讨其细胞内谷胱甘肽(GSH)水平.方法:As2O3和AA不同组合孵育细胞HepG2,采用台盼蓝染色实验检测药物对细胞生长的影响,流式细胞仪检测亚二倍体凋亡峰,Annexin V染色检测细胞凋亡率,GSH实验检测细胞内GSH水平.结果:100 μmol/L的AA单独作用与对照组相比在细胞增殖和凋亡都无显著性差异;低剂量(2 μmol/L)和高剂量(5 μmol/L)的As2O3都能抑制细胞增殖活性和诱导细胞凋亡,联合AA作用后,比相应浓度的单独As2O3作用更为显著;AA和As2O3分别处理都能明显消减细胞GSH含量,两种药物联合作用后与相应单独As2O3作用相比GSH水平显著性降低.结论:GSH在As2O3对肝癌细胞抑制增殖和诱导凋亡作用机制中起到重要的作用,高低剂量的As2O3联合AA通过降低GSH水平均可以增强As2O3对肝癌细胞抑制生长及其诱导凋亡作用的敏感性.     

三氧化二砷影响肝癌细胞血管内皮生长因子表达信号转导机制的初步探讨

目的：探讨三氧化二砷（As2O3）影响人肝癌细胞株HepG2表达血管内皮生长因子（VEGF）的信号转导机制。方法：体外培养肝癌细胞系HepG2,实验分为：①空白组、②对照（As2O3）组、③As2O3＋PKC阻断剂（H7）组、④As2O3＋P38阻断剂（SB203580）组、⑤As2O3＋H7＋SB203580组;半定量RT—PCR技术检测HepG2细胞VEG-FmRNA相对表达量。结果：②、③、④、⑤纽均有VEGF的表达,与①组相比差异有统计学意义（P〈0．01）,③、④组与②组相比无差异;⑤组与②组相比VEGF的表达明显降低,P〈0．05;结论：三氧化二砷可诱导HepG2细胞表达VEGFmRNA,并可能通过PKC和／或P38信号通路影响VEGFmRNA的表达。     

维甲酸联合三氧化二砷对NB4细胞NF-κB、ROS表达的影响

摘要目的探讨全反式维甲酸(ATRA)联合三氧化二砷(As2O3)对NB4细胞的生长以及核因子-κB(NF-κB)、活性氧(ROS)表达的影响.方法选择NB4细胞,加入ATRA和不同剂量的As2O3,观察细胞生长曲线,流式细胞仪检测CD11 b、ROS、NF-κB的变化,DNA片段梯度观察细胞凋亡.结果 0.5 μmol/L As2O3下调了1 μmol/L ATRA诱导的NB4细胞的分化凋亡发生,同时抑制了NF-κB的激活,提高了ROS水平.2 μmol/L As2O3联合同浓度ATRA对增殖抑制、诱导凋亡更明显.结论 ATRA联合As2O3对NB4细胞具有分化和凋亡的双重效应.     

三氧化二砷诱导人肝癌HepG2细胞凋亡及细胞周期阻滞的研究

目的 研究三氧化二砷(arsenic trioxide,As2O3)对人肝癌HepG2细胞的生长抑制作用,以及诱导细胞凋亡和对周期分布的影响.方法 采用MTF法、DNA ladder检测、流式细胞术等方法 检测As2O3对人肝癌HepG2细胞的生长抑制,诱导细胞凋亡以及对细胞周期分布的影响.结果 MTT法显示:As2O3对HepG2细胞有明显的生长抑制作用,且具有剂量和时间依赖性.12μmol/L As2O3作用HepG2细胞24、48、72 h后,DNA ladder检测,48、72 h均出现凋亡典型的DNA ladder条带;流式细胞仪分析显示:As2O3处理HepG2细胞24、48、72 h后,凋亡率分别为(9.74±1.22)%、(21.89±2.14)%、(42.72±0.83)%,均显著高于对照组细胞的凋亡率(0.98±0.11)%(P<0.01).同时,流式细胞仪分析显示,As2O3对HepG2细胞有明显的S期和G2期阻滞作用.结论 As2O3对人肝癌HepG2细胞S期和G1期阻滞并诱导细胞凋亡可能是其抗肿瘤的机制之一.     

三氧化二砷对恶性黑色素瘤A375细胞株凋亡的诱导作用

目的:研究三氧化二砷(As2O3)对人恶性黑色素瘤A375细胞株凋亡的诱导作用.方法:采用流式细胞术(FCM)检测不同浓度的三氧化二砷诱导A375细胞的早期凋亡率和晚期凋亡率.结果:As2O3浓度分别为5 μmol/L、10μmol/L、20 μmol/L时,A375细胞的早期凋亡率分别为9.75%、50.9%和43.7%,晚期凋亡率分别为2.10%、4.61%和11.2%,总凋亡率分别为11.85%、55.51%和54.9%.结论:As2O3可诱导A375细胞早期凋亡和晚期凋亡,10 μmol/L和20μmol/L的As2O3可明显增加A375细胞总凋亡率.     

细胞分化剂增强三氧化二砷诱导肝癌细胞的凋亡效应

【目的】探讨细胞分化剂 (CDA Ⅱ )在三氧化二砷 (As2 O3 )诱导肝癌细胞凋亡效应中的作用。【方法】应用CDA Ⅱ和As2 O3 共同处理肝癌细胞株BEL 74 0 2、HepG2 ,通过四唑蓝比色法检测细胞存活率 ;用活细胞荧光染色观测细胞凋亡及形态学变化、流式细胞术分析细胞周期变化及凋亡率。【结果】CDA Ⅱ毒性作用很低 ,但可以显著增强As2 O3 对肝癌细胞生长的抑制作用 ,1 0 g/LCDA Ⅱ便可使As2 O3 对两株细胞的半数抑制浓度由 5 0 μmol/L降低至 1 0 μmol/L(P <0 0 1)。形态学可观察到CDA Ⅱ明显加强了As2 O3 诱导的细胞凋亡 ,且与剂量有关 ;流式细胞术分析 ,低质量浓度的CDA Ⅱ (<2 0 g/L)细胞生长阻滞于G2 期较对照组多 ,而随着剂量的增加则凋亡细胞和滞留于G1期的细胞增多 ,低剂量CDA Ⅱ与As2 O3 共同处理肝癌细胞后 ,凋亡率明显高于单独用As2 O3 。【结论】CDA Ⅱ可增强As2 O3 诱导肝癌细胞的凋亡效应 ,两药具有协同作用。     

三氧化二砷对鼠恶性黑素瘤B16细胞凋亡的影响

目的:探讨三氧化二砷(As2O3)对鼠恶性黑素瘤B16细胞诱导凋亡的影响.方法:采用流式细胞术(FCM)检测不同浓度的As2O3诱导B16细胞的早期凋亡率、晚期凋亡率和总凋亡率.结果:As2O3浓度分别为5 μmol/L、10 μmol/L 、20 μmol/L时, B16细胞的早期凋亡率分别为7.18%、21.1%、3.59%,晚期凋亡率分别为2.63%、5.35%、6.34%,总凋亡率分别为9.81%、26.45%、9.93%.结论: B16细胞的早期凋亡率和晚期凋亡率对As2O3有浓度依赖关系,高浓度的As2O3使B16细胞出现坏死.     

三氧化二砷与顺铂联用对肝癌HepG2细胞的作用

目的 探讨三氧化二砷(As2O3)与顺铂(CDDP)联合对肝癌细胞HepG2的作用.方法 应用MTT法和细胞免疫组化酶法检测人肝癌HepG2细胞Bax,Bcl-2,Fas蛋白的表达情况.结果 As2O3与CDDP联合应用在抑制肝癌细胞的生长繁殖、诱导肝癌细胞凋亡,增强Bax,Fas的表达,下调Bcl-2的表达方面均较各自单用的作用明显增强.结论 As2O3与CDDP联合应用具有明显的协同抗肝癌作用,其主要机制可能为As2O3联合化疗能诱导肝癌细胞凋亡,与增强Bax,Fas的表达,下调Bcl-2的表达有关.     

抗坏血酸与三氧化二砷联合应用诱导肝癌细胞凋亡

目的探讨抗坏血酸(ascorbic acid,AA)与三氧化二砷(As2O3)联合应用增强肝癌细胞凋亡的作用。方法肝癌细胞株BEL-7402及SMMC-7721细胞分别用As2O3(1 μmol/L)、AA(62.5 μmol/L)、As2O3(1 μmol/L) AA(62.5 μmol/L)处理24～96 h,流式细胞仪上检测细胞凋亡百分率;用试剂盒法测定细胞内谷胱甘肽(glutathione, GSH)含量。结果1 μmol/As2O3作用24～96 h有诱导BEL-7400及SMMC-7721细胞凋亡的作用,从(62.5 μmol/L) As2O3(1 μmol/L)作用72 h BEL-7402细胞凋亡百分率为(61.7±0.8)%,SMMC-7721细胞为(26.8±0.9)%,较单独应用As2O3(1 μmol/L)的凋亡百分率[(40.2±0.8)%;(11.4±0.5)%]明显增高。As2O3(1 μmol/L)单独作用对SMMC-7721和BEL-7402细胞内GSH的含量无明显影响。从(62.5 μmol/L)单独作用对BEL-7402细胞内GSH的含量无明显影响,但可明显降低SMMC-7721细胞内GSH的含量(P<0.05)。AA(62.5 μmol/L)与As2O3(1 μmol/L)联合作用可明显降低SMMC-7721和BEL-7402细胞内GSH的含量(P<0.05),SMMC-7721细胞内GSH的含量降低更显著(P<0.01)。结论AA及As2O3联合应用能增强肝癌细胞株BEL-7402及SMMC- 7721细胞凋亡作用,尤其是对As2O3不敏感细胞株SMMC-7721细胞作用更显著。     

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.