三氧化二砷对胃癌细胞放射增敏作用的研究

目的探讨三氧化二砷（As2O3）是否对胃癌细胞有放射增敏作用。方法以人胃癌细胞株SGC-7901为研究对象,所有实验均重复3次,取其均值为最终结果。①四甲基四氮唑盐比色法（MTT法）检测As2O3的单药毒性,确立细胞半数抑制浓度（IC50）,并行单纯照射组及As2O3＋照射组MTT检测,计算q值大小。②放射增敏实验分成空白对照组、单纯给药组、单纯照射组及As2O3＋照射组,采用平板克隆形成分析法计算各组细胞存活分数（SF）,运用＂多靶单击数学模型＂拟合细胞存活曲线,得出平均致死剂量（Do）、准阈剂量（Dq）、放射增敏比（SER）,并观察单纯照射组及As2O3＋照射组对肿瘤细胞的杀伤作用。结果胃癌细胞株SGC-7901细胞IC50值为12.65μmol/L,与单纯照射组及单纯给药组相比,As2O3与放射治疗联合作用能显著抑制SGC-7901细胞的增殖,并显示出As2O3与放射治疗的协同作用（q〉1.15）。单纯照射组Do、Dq分别为2.75、2.52Gy,2μmol/LAs2O3＋照射组Do、Dq、SER（Do值比）、SER（Dq值比）分别为2.18Gy、1.89Gy、1.26、1.33,5μmol/LAs2O3＋照射组Do、Dq、SER（Do值比）、SER（Dq值比）分别为1.78Gy、1.72Gy、1.54、1.51。结论As2O3对人胃癌细胞株SGC-7901有一定的放射增敏作用,为临床放疗及与As2O3联合运用提供了实验依据。     

三氧化二砷对乳癌细胞系MDA-MB-468雌激素受体表达的影响

目的:探讨三氧化二砷(As2O3)对人雌激素受体α(ERα)阴性的乳癌细胞株MDA-MB-468进行药物诱导后ERα启动子CPG岛的甲基化状态及蛋白的变化。方法:常规培养MDA-MB-468细胞,采用MTT法检测0.5、1.0、2.0、4.0、8.0和16.0mmol/LAs2O3分别处理24、48和72h后细胞的生长抑制率,甲基化特异性PCR检测1.0、2.0、4.0μmol/LAs2O3处理72h后MDA-MB-468细胞ERα启动子CpG岛的甲基化状态,Westernblot检测1.0、2.0、4.0μmol/LAs2O3处理72h后MDA-MB-468细胞ERα蛋白的表达情况。结果:As2O3可抑制MDA-MB-468细胞增殖,其作用呈时间和剂量依赖性(F浓度=375.603,F时间=474.827,P<0.001)。经1.0、2.0、4.0μmol/LAs2O3处理后的MDA-MB-468细胞启动子CpG岛甲基化水平降低,ERα蛋白重新表达。结论:As2O3明显抑制MDA-MB-468增殖,适当浓度As2O3能诱导ERα去甲基化,使ERα阴性的MDA-MB-468细胞恢复表达ERα。     

As2O3与阿司匹林联合应用增强对人胃癌SGC-7901细胞增殖的抑制作用

目的:探讨As2O3与阿司匹林联合应用对人胃癌SGC-7901细胞生长的影响。方法:采用MTT法检测As2O3和阿司匹林对SGC-7901细胞株增殖的影响,采用流式细胞术检测两者对SGC-7901细胞凋亡和细胞周期分布的影响。结果:①药物作用后72h,4μmol/L和2mmol/LAs2O3组生长抑制率分别为31.04%及16.11%,与阴性对照组相比差异具有统计学意义(P<0.05);2μmol/LAs2O3 1mmol/L阿司匹林联合用药组的生长抑制率为24.19%,与阴性对照组、2μmol/LAs2O3(5.96%)及1mmol/L阿司匹林(4.86%)相比,差异均有统计学意义(P<0.05);②2μmol/LAs2O3与1mmol/L阿司匹林联合组细胞的凋亡率是19.01%,与阴性对照组(1.55%)、2μmol/LAs2O3组(1.81%)及1mmol/L阿司匹林组(1.64%)相比,差异均有统计学意义(P<0.05);③As2O3使细胞G0/G1期比例下降,G2/M期比例增加。阿司匹林使细胞G0/G1期比例增加,S和G2/M期比例下降。2μmol/LAs2O3与1mmol/L阿司匹林联合应用后72h,G0/G1期和G2/M期比例增加,S期比例下降。结论:As2O3、阿司匹林可以抑制SGC-7901细胞增殖、诱导其凋亡,阻滞细胞周期,两者联合应用具有协同作用,可以同时降低两药的用量。     

电离辐射和三氧化二砷联合对人脑胶质瘤SHG44细胞的杀灭作用

目的 研究电离辐射联合三氧化二砷(As2O3)对人脑胶质瘤SHG44细胞的杀灭作用。方法 以SHG44细胞为实验对象,用四氮唑盐(MTT)比色法观察不同剂量的电离辐射、单独As2O3及不同剂量的电离辐射和As2O3联合对SHG44细胞的杀灭作用;用激光共聚焦显微镜观察细胞的形态学变化。结果 (1)照射与As2O3联合组SHG44细胞存活率低于单独相应剂量照射组(P<0.01)和单独As2O3组(P<0.01);(2)2Gy剂量照射+4μmol／L As2O3组对SHG44细胞杀灭作用强于6Gy剂量单独照射组(P<0.01),相当于8Gy单独照射组(P>0.05);(3)激光共聚焦显微镜下观察到As2O3处理组细胞出现明显的凋亡形态学变化。结论 电离辐射和As2O3联合对SHG44细胞的杀灭作用分别强于电离辐射和As2O3单独作用;As2O3对SHG44细胞的杀灭作用机制主要     

三氧化二砷对人大肠癌细胞生长的抑制作用

目的探讨不同浓度的三氧化二砷对人大肠癌细胞株CCL-187细胞生长抑制率的影响。方法大肠癌细胞株CCL-187分别与1·0,2·5μmol/LAs2O3共同孵育1~6d,电镜下观察细胞形态学变化,MTT法计算细胞生长抑制率。结果MTT法显示1·0,2·5μmol/LAs2O3均能抑制大肠癌CCL-187细胞的生长增殖,且具有时间依赖性。As2O3对CCL-187细胞增殖的抑制程度随As2O3浓度的加大而加强。透射电镜显示:1·0μmol/LAs2O3促进细胞分化,但2·5μmol/LAs2O3能诱导细胞凋亡。结论不同浓度的三氧化二砷对人大肠癌CCL-187细胞生长均可产生抑制作用,作用机制为1·0μmol/LAs2O3促进癌细胞分化,而2·5μmol/LAs2O3则诱导细胞凋亡。     

VPA联合As2O3对NB4细胞VEGF及Survivin表达的影响

目的:探讨丙戊酸钠(VPA)联合三氧化二砷(As2O3)对NB4细胞株血管内皮生长因子(VEGF)、凋亡抑制蛋白Survivin表达的影响。方法:NB4细胞分为对照组、As2O3组及As2O3联合VPA组,RT-PCR方法检测NB4细胞VEGF、Survivin mRNA的表达。结果:As2O30.1μmol/L组NB4细胞VEGF mRNA的表达明显高于对照组、Survivin mRNA的表达明显低于对照组(P<0.05);与As2O30.1μmol/L组比较,As2O30.1μmol/L+VPA 1.0mmol/L组NB4细胞VEGF mRNA、Survivin mRNA的表达均明显降低(P<0.05)。结论:VPA联合小剂量As2O3可以降低单独应用As2O3导致的NB4细胞VEGF表达的升高、并进一步降低Survivin的表达,As2O3联合VPA可降低As2O3治疗急性早幼粒细胞白血病时的不良反应。     

蛋白酶体抑制剂联合亚砷酸对NB4细胞survivin基因表达的影响

目的 研究蛋白酶体抑制剂硼替佐米(bortezomib)单药应用及其联合亚砷酸(As2O3)对急性早幼粒细胞白血病(APL)细胞株NB4中survivin mRNA表达的影响.方法 将受试对象分为9组:未加药空白对照组(D组);As2O3组,0.5 μmol/L(A1组)和1μmol/L(A2组);bortemmib组,5 nmol/L(B1组)和10 nmol/L(B2组);联合组,0.5/μmol/L As2O3 5 nmol/Lbortezomib(C1组),0.5μmol/L As2O3 10 nmol/L bortezomib(C2组),1μmol/L As2O3 5 nmol/L bortezomib(C3组),1μmol/LAs2O3 10 nmol/L bortezomib(CA组).作用时间分为三个水平:24 h,48 h和72 h.应用逆转录聚合酶链反应(RT-PCR)法检测药物作用前后抑凋亡基因survivin mRNA的表达水平.结果 bortezomib和As2O3单药组中survivin mRNA的表达水平下降,且存在剂量与时间依赖性,联合组survivin mRNA的表达水平亦下降,且比单药组下降更明显.结论 bortezomib与As2O3可通过抑制NB4细胞株survlvin mRNA的表达诱导细胞发生凋亡,两药联合具有协同效应.     

丙戊酸钠联合三氧化二砷对HL-60细胞株的体外研究

目的:探讨丙戊酸钠(sodium valproate,SV)联合三氧化二砷(arsenous trioxide,As2O3)诱导HL-60细胞凋亡与bcl-2﹑bax mRNA之间可能的关系。方法:在对数生长期的HL-60细胞中分别加入不同浓度的SV和1.0μmol/LAs2O3,采用MTT比色法测定细胞生长抑制率,用Annexin V-FITC/PI双重标记经流式细胞仪检测细胞早期凋亡率,RT-PCR检测bcl-2,bax mRNA在细胞中的表达。结果:SV抑制HL-60细胞生长并促进其凋亡,SV联合1.0μmol/L As2O3对HL-60细胞的生长抑制及促凋亡作用更明显。SV单独或联合1.0μmol/L As2O3作用于HL-60细胞,bcl-2mRNA表达下降,bax mRNA表达增高。结论:SV与As2O3有协同作用,抑制HL-60细胞生长并诱导其凋亡,bcl-2﹑bax mRNA的表达变化可能参与了SV对HL-60细胞的促凋亡作用。     

半乳糖受体介导的c-myc反义核酸联合三氧化二砷对人肝癌细胞的增殖抑制作用

目的　探讨Gal-PEI-c -mycASODN与三氧化二砷 (As2 O3 )联合应用对人肝癌Bel- 74 0 2细胞增殖的影响。方法　As2 O3 分别或联合Gal-PEI-c -mycASODN与人肝癌Bel- 74 0 2细胞共孵育 4 8h ,台盼蓝染色法检测细胞增殖的变化 ,流式细胞仪分析细胞亚二倍体百分率。结果　 0 5 μmol/LAs2 O3 和Gal-PEI-ASODN(含A SODN 0 12 5 μmol/L)均不能抑制Bel- 74 0 2细胞增殖 ,0 5 μmol/L的As2 O3 联合Gal-PEI-ASODN(含ASODN 0 12 5μmol/L)对Bel- 74 0 2细胞增殖有明显抑制作用 (P <0 0 5 ) ,与 4 μmol/LAs2 O3 产生的抑制作用相当 ;1 0 μmol/LAs2 O3 不引起Bel- 74 0 2细胞凋亡 ,但联合Gal-PEI-ASODN(含ASODN 0 2 5 μmol/L)即导致Bel - 74 0 2细胞凋亡。结论　Gal-PEI-ASODN联合As2 O3 能明显提高肝Bel- 74 0 2细胞对As2 O3 的敏感性 ,抑制Bel- 74 0 2细胞增殖 ,促进细胞凋亡。     

三氧化二砷对人骨髓增生异常综合征MUTZ-1细胞的作用

目的:研究不同浓度三氧化二砷(As2O3)对人骨髓增生异常综合征难治性贫血伴原始细胞过多型(MDS-RAEB)细胞株MUTZ-1细胞增殖的影响,并探讨其可能的作用机制。方法:采用MTT法分析细胞生长曲线,用细胞形态学观察细胞凋亡。结果:低浓度As2O3(0.05～0.25μmol/L)对MUTZ-1细胞无明显生长抑制作用,高浓度As2O3(1.0～20.0μmol/L)对MUTZ-1细胞有明显生长抑制作用,且呈浓度、时间效应-依赖关系(P<0.01)。结论:低浓度As2O3(0.05～0.25μmol/L)对MUTZ-1细胞无明显生长抑制作用。高浓度As2O3(1.0～20.0μmol/L)对MUTZ-1细胞有明显生长抑制作用,且呈浓度、时间效应依赖关系。     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

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.     

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.