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。     

As_2O_3联合~（89）SrCl_2对人乳腺癌细胞株MCF-7细胞周期和凋亡的影响

目的探讨三氧化二砷（As2O3）联合氯化锶（89SrCl2）对人乳腺癌细胞株MCF-7细胞周期与凋亡的影响。方法采用MTT比色法检测As2O3对MCF-7细胞的增殖抑制作用,求出细胞半数抑制浓度（ID50）。选择20%ID50以下两个不同浓度的As2O3联合89Sr照射,随机设置对照组、89SrCl2照射组、As2O3处理组,As2O3＋89SrCl2联合处理组（联合组）,并于处理后24h采用流式细胞术检测各组细胞周期分布及凋亡。结果 As2O3能明显抑制MCF-7细胞增殖,给药24h的ID50为11.7μmol/L。细胞流式术检测结果表明,与89SrCl2照射组相比,联合组G2/M期细胞明显增多（P〈0.05或0.01）,早期凋亡和死亡细胞数显著增高（P〈0.05）。结论 As2O3能够促进89Sr照射诱导的MCF-7细胞周期阻滞与凋亡。     

砷剂对乳腺癌MCF-7和MCF-7/ADR 细胞生长抑制作用的比较研究

目的:探讨As2O3对人乳腺癌MCF-7细胞和多药耐药MCF-7/ADR细胞生长抑制作用的差异.方法:采用MTI还原法、光镜、电镜和流式细胞术等方法检测As2O3对MCF-7和MCF-7/ADR细胞的生长抑制作用和诱导凋亡的情况.结果:As2O3对MCF-7和MCF-7/ADR细胞均有生长抑制作用,并呈时间剂量依赖关系,96h的IC50值分别为3μmol/L和12.8μmol/L,形态学检测显示其抑制作用主要源于凋亡;在相同作用时间及浓度下,As2O3对MCF-7细胞的抑制率显著高于MCF-7/ADR细胞,起效时间亦早于后者.结论:As2O3能诱导乳腺癌MCF-7和MCF-7/ADR细胞凋亡;MCF-7/ADR细胞虽对As2O3表现耐药,但耐药倍数较低(4.27).     

三氧化二砷对乳腺癌细胞株的抑制作用及其机制探讨

目的 探讨三氧化二砷(arsenous oxide,As2O3)对乳腺癌MCF-7细胞的抑制作用及其机制.方法 对乳腺癌MCF-7细胞株进行培养传代.As2O3作用于细胞一定时间后利用流式细胞仪检测乳腺癌细胞凋亡率;逆转录-聚合酶链反应(RT-PCR)检测survivin基因的表达;电镜观察乳腺癌细胞凋亡情况.结果 As2O3对乳腺癌细胞的抑制作用是以促进细胞凋亡为主;凋亡抑制因子survivin基因在乳腺癌细胞中表达;乳腺癌细胞经As2O3作用后,能抑制survivin基因的表达.结论 As2O3可能通过抑制凋亡抑制因子survivin基因的表达,促进细胞凋亡,从而发挥抗肿瘤的作用.     

三氧化二砷抑制人乳腺癌细胞生长及其作用机制的初步研究

目的观察三氧化二砷(As2O3)对人乳腺癌MDA MB231细胞的生物学效应及其作用机制。方法采用MTT法观察细胞毒性;膜联蛋白V(AnnexinV)异硫氰酸荧光素(FITC)+碘化丙啶(PI)双参数检测细胞凋亡;流式细胞术测定细胞周期、增殖细胞核抗原(PCNA)、凋亡相关蛋白Fas和bcl2阳性细胞百分率及细胞内钙离子(IECa2+)含量变化。结果As2O3可显著抑制MDA MB231细胞生长,且剂量效应关系差异有统计学意义(r=0.99,P<0.01),其IC50为6.65μmol/L;MDA MB231细胞经As2O3处理后可观察到凋亡;As2O3能上调Fas蛋白表达和IECa2+含量(P<0.01),下调PCNA蛋白表达(P<0.01),并使细胞周期阻滞在S+G2/M期,但对bcl2表达无影响(P>0.05)。结论As2O3在体外可显著抑制MDA MB231细胞生长并引起凋亡,其机制可能与上调Fas表达和IECa2+含量、降低PCNA表达及阻滞细胞周期有关。     

姜黄素对人乳腺癌MCF-7细胞增殖抑制作用及机制研究

目的:探讨姜黄素对人乳腺癌MCF-7细胞增殖抑制、诱导凋亡作用及其分子作用机制。方法:MTT法检测姜黄素对MCF-7细胞的增殖抑制作用;流式细胞术PI单染法检测细胞周期;Annexin V/PI双染法检测细胞凋亡;RT-PCR法检测凋亡相关基因Bcl-2和Bax的mRNA表达水平。结果:姜黄素对MCF-7细胞生长有明显抑制作用,并呈剂量、时间依赖性;FCM结果显示姜黄素能使MCF-7细胞阻滞在G1/S期;Annexin V/PI双染法验证姜黄素可以诱导细胞凋亡;RT-PCR结果显示Bax mRNA水平明显上调,而Bcl-2的mRNA表达水平降低。结论:姜黄素对人乳腺癌MCF-7细胞增殖具有显著的抑制作用并可诱导细胞凋亡,其作用机制可能与其上调Bax基因表达水平的同时,下调Bcl-2基因表达水平,从而诱导细胞凋亡有关。     

三氧化二砷诱导喉癌Hep—2耐药细胞凋亡的研究

目的 :观察三氧化二砷 (As2 O3 )对喉癌Hep -2细胞株和长春新碱诱导的多药耐药Hep -2r细胞的作用和对细胞周期的影响。方法 :用长春新碱(VCR)递增药物浓度法筛选耐药细胞Hep -2r,体外培养的细胞与不同浓度的As2 O3 作用 2 4、48、72h,通过MTT还原法检测细胞的生长抑制率 ,用光学显微镜、流式细胞仪、荧光显微镜研究细胞凋亡的形态学改变 ,检测细胞凋亡率并进行细胞周期分析。结果 :As2 O3 可有效抑制Hep -2细胞和Hep-2r细胞的生长 ,呈时间和浓度依赖性特点。形态学观察发现 ,As2 O3 诱导Hep-2和Hep-2r细胞凋亡 ,Hep -2和Hep -2r细胞对As2 O3 的敏感性无显著差异。2 μmol/LAs2 O3 作用 2 4h时 ,S期细胞比例增高 ,72h后 ,S期细胞明显下降 ,细胞大量凋亡。As2 O3 在作用早期 ,阻滞细胞通过S期 ,随着时间的延长 ,诱导S期细胞凋亡。结论 :As2 O3 可诱导Hep -2细胞和Hep-2r细胞凋亡 ,与细胞周期阻滞有关     

三氧化二砷诱导喉癌Hep-2细胞凋亡及其对细胞周期的影响

目的 观察三氧化二砷（AS2O3）对喉癌Hep-2细胞株诱导细胞凋亡的作用和对细胞周期的影响。方法 体外培养的Hep-2细胞与不同浓度的AS2O3作用24、48、72h，通过MTT还原法检测细胞的生长抑制率，用倒置相差显微镜、流式细胞仪、荧光显微镜研究细胞凋亡的形态学改变，检测细胞凋亡率并进行细胞周期分析。结果 As2O3可有效抑制Hep-2细胞的生长，呈时间和浓度依赖性特点。形态学观察发现，As2O3诱导Hep-2细胞凋亡。2μmol/L As2O3作用24h，S期细胞比例增高，72h后，S期细胞明显下降，细胞大量凋亡。AS2O3在低浓度时，阻滞细胞通过S期，高浓度时诱导S期细胞凋亡。结论 AS2O3可诱导Hep-2细胞的凋亡，与细胞周期阻滞有关。     

三氧化二砷诱导人卵巢癌耐药细胞系3AO/cDDP细胞凋亡及机制的研究

目的：探讨三氧化二砷（As2O3）对人卵巢癌耐药细胞系3AO／cDDP细胞生长的影响及其机制。方法：采用四甲基偶氮唑蓝（MTT）法,检测不同浓度As2O3作用后,3AO／cDDP细胞的生长抑制率;采用流式细胞技术检测细胞凋亡率,细胞周期变化,以及Fas、FasL基因表达的变化。所有结果均与人卵巢癌细胞系3AO细胞相比较;采用细胞骨架染色法观察3AO／cDDP细胞凋亡细胞形态变化,通过吖啶橙染色,荧光显微镜观察As2O3作用后3AO细胞的形态变化。结果：As2O3能明显抑制3AO／cDDP细胞的增殖,抑制作用呈时间和剂量依赖性（P＜0．05）;在定一浓度范围内,3AO／cDDP细胞凋亡率与As2O3的浓度和作用时间呈依赖关系,诱导凋亡的最适浓度是3μmol/L;As2O3低浓度时,3AO／cDDP细胞周期S期通过受阻,高浓度时诱导S期细胞凋亡;As2O3作用后,两种细胞系Fas基因的表达均呈升调节,FasL基因的表达无变化;与3AO细胞相比,差异无显著意义（P＞0．05）;As2O3作用后3AO及3AO／cDDP形成典型的凋亡小体。结论：As2O3通过Fas/FasL系统,诱导S期细胞凋亡,有效地抑制人卵巢癌耐药细胞系细胞的生长。     

Nicotiflorin对人乳腺癌MCF-7细胞增殖及凋亡的影响

目的：探讨Nicotiflorin（Kaempferol3-O-β-rutinoside,山奈酚-3-O-芸香糖苷）对人乳腺癌MCF-7细胞的增殖抑制,以及诱导细胞凋亡和对细胞周期的影响。方法采用MTS法检测细胞存活率,计算IC50值;流式细胞仪检测细胞周期及细胞凋亡;Western-blot检测DNA损伤标记蛋白γ-H2AX及凋亡相关蛋白Bcl-2的变化。结果Nicotiflorin能剂量依赖性地抑制人乳腺癌MCF-7细胞的增殖,IC50值为28．2μmol/L。流式细胞仪检测结果显示,Nicotiflorin使MCF-7细胞周期阻滞于G2/M期,不同浓度的Nicotiflorin作用48h后细胞凋亡率均增加。Western-blot检测发现,Nicotiflorin处理后,磷酸化的H2AX（γ-H2AX）随时间依赖性地增加,提示Nicoti-florin诱导DNA双链断裂从而使细胞阻滞于G2/M期,同时Nicotiflorin可能通过抑制抗凋亡蛋白Bcl-2的表达诱导细胞凋亡。结论Nicotiflorin能抑制人乳腺癌MCF-7细胞的增殖并诱导其凋亡。     

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.