藤黄酸诱导膀胱癌细胞凋亡和细胞周期阻滞作用

目的：藤黄酸能抑制包括乳腺癌、肝癌、血液系肿瘤以及乳腺癌在内的多种肿瘤细胞的生长,但是目前对于藤黄酸在泌尿系肿瘤作用的报道尚少。文中旨在研究藤黄酸诱导膀胱癌细胞凋亡和细胞周期阻滞作用可能的机制。方法实验分为4组,藤黄酸1．0μmol／L组、2．0μmol／L组、3．0μmol／L组及阴性对照组（加入等渗盐水）。体外培养人膀胱癌细胞株BIU-87,将不同浓度藤黄酸加入处于对数生长期的人膀胱癌BIU-87细胞共培养,免疫组化法检测瘤组织Caspase-3蛋白的表达,使用流式细胞仪检测分析藤黄酸诱导人膀胱癌细胞的凋亡作用和细胞周期变化。结果藤黄酸1．0μmol／L组、2．0μmol／L组、3．0μmol／L组免疫组化评分分别为（4．28±1．86、5．03±0．78和6．47±1．31）,较对照组（2．13±1．27）明显升高,差异均有统计学意义（ P＜0．05）。流式细胞术结果显示：细胞周期 G0／G1期细胞逐渐减少,G2／M 期细胞逐渐增多,S期细胞变化不明显。结论藤黄酸可通过增加 Caspase-3蛋白的表达来促进膀胱癌细胞凋亡,可阻滞膀胱肿瘤细胞周期,抑制膀胱肿瘤细胞的增殖。     

榄香烯联合热疗诱导膀胱癌BIU-87细胞凋亡及对p-Akt表达的影响

目的探讨榄香烯联合热疗诱导人膀胱癌细胞株-87（BIU-87）膀胱癌细胞凋亡及对磷酸化丝氨酸苏氨酸蛋白激酶（p-Akt）蛋白表达的影响。方法采用流式细胞仪和透射电子显微镜分析和观察不同浓度榄香烯与热疗单独或联合对BIU-87膀胱癌细胞作用结果;Westernblotting检测榄香烯（40mg／L）与热疗单独或联合作用于人BIU膀胱癌细胞24h后p-Akt蛋白的表达。结果榄香烯联合热疗处理后BIU-87细胞超微结构出现明显的凋亡形态学改变,不同浓度的榄香烯组、热疗组和榄香烯联合热疗组作用于人BIU-87膀胱癌细胞24h均可使细胞凋亡率增加,但榄香烯联合热疗组的作用显著强于前两者;凋亡率增加的同时下调了p-Akt蛋白的表达。结论榄香烯联合热疗可诱导人BIU-87膀胱癌细胞凋亡,且过程中下调了p-Akt蛋白的表达。     

PTEN基因转染对人膀胱癌细胞BIU-87凋亡的影响

目的：研究外源性抑癌基因PTEN转染对人膀胱癌细胞系BIU-87凋亡的影响,探索膀胱癌基因治疗的新途径.方法：将携有PTEN基因的真核表达载体体外转染BIU-87细胞,筛选稳定转染的细胞并扩增培养,用逆转录聚合酶链反应法（RT-PCR）检测PTEN的表达情况,应用细胞原位凋亡检测试剂盒和流式细胞仪分别研究PTEN基因转染前、后膀胱癌细胞凋亡的变化.结果：转染PTEN基因的膀胱癌细胞系BIU-87中PTEN mRNA能稳定表达,肿瘤细胞凋亡明显增多,与对照组比较有显著性差异.结论：转染外源性PTEN基因能显著诱导膀胱癌细胞凋亡,从而达到抑制肿瘤发生、发展的目的.     

膀胱移行细胞癌中粘蛋白MUC1的表达与肿瘤浸润性树突状细胞的分布

目的研究膀胱移行细胞癌中粘蛋白MUC1表达和肿瘤浸润性树突状细胞（TIDC）分布的变化，探讨二者在膀胱癌侵袭、转移及化疗耐药过程中的作用。方法采用免疫组织化学SP法检测MUC1和TIDC在膀胱移行细胞癌的表达和定位及MUC1在T-24细胞、BIU-87和耐药株BIU-87／A中的表达。应用流式细胞仪检测BIU-87、耐药株BIU-87／A及T-24细胞在阿霉素、长春新碱、顺铂3种化疗药物作用48h后的凋亡率：结果MUC1在各期膀胱移行细胞癌中均表达，表达模式各有特点，MUC1的染色分型同肿瘤的病理分级和临床分期密切相关（P〈0．001）。DC的数量同肿瘤病理分级呈负相关．随着病理分级的升高阳性细胞数明显减少（P〈0．005）。MUC1存BIU-87、T-24细胞膜、胞浆中均有浅棕色弱阳性表达，在BIU-87／A的胞膜、胞浆中均有深棕色强阳性表达，两者细胞平均光密度值差异有显著性（P〈0．01）。流式细胞仪检测BIU-87、T-24和BIU-87／A的自发凋亡率分别为1．15％、1．40％、0．90％，在阿霉素、长春新碱、顺铂3种化疗药物作用48h后．BIU-87的凋亡率分别为45．69％、47．70％、44．50％，T-24的凋亡率分别为43．79％、46．17％、44．50％，均有明显升高（P〈0．01），但两种细胞之间及3种化疗药之间差异无显著性（P〉0．05）。耐药株BIU-87／A在阿霉素、长春新碱作用48h后的凋亡率分别为19．88％、21．41％．均明显低于亲本细胞株BIU-87（P〈0．05）。结论MUC1的表达模式和TIDC数量的监测可以作为膀胱移行细胞癌恶性程度和预后的判断指标。TIDC的减少可能是膀胱癌免疫逃逸和耐受的重要环节，肿瘤细胞表面高密度的MUC1可能参与膀胱移行细胞癌侵袭转移和化疗耐药机制。     

芹菜素诱导膀胱癌细胞株BIU-87凋亡及机制的研究

目的:探讨芹菜素(Apigenin)在体外诱导膀胱癌细胞株BIU-87凋亡的作用及其机制.方法:MTT法测定不同浓度芹菜素作用后BIU-87细胞的生长情况;流式细胞术检测细胞周期情况;Annexin V-FITC/PI双染流式细胞术观察芹菜素作用后细胞的凋亡情况;RT-PCR法检测Survivin mRNA表达的变化;免疫细胞化学SP法检测Survivin和Caspase-3蛋白表达的变化.结果:①MTT法证实芹菜素能明显抑制膀胱癌细胞株BIU-87的生长.②22.03 μmol/L芹菜素作用72 h后,流式细胞仪检测发现芹菜素可使BIU87细胞阻滞在G2/M期;流式细胞术双染法检测细胞凋亡显示:芹菜素可使BIU-87细胞发生凋亡.③RT-PCR检测发现芹菜素可使BIU-87细胞Survin mRNA表达显著减少(P<0.05),免疫细胞化学SP法检测显示Survivin蛋白表达亦显著减少(P<0.05),Caspase-3蛋白表达则明显增加(P<0.01).结论:芹菜素在体外可抑制BIU-87细胞增殖,诱导其发生凋亡.芹菜素诱导BIU-87细胞发生凋亡的生物学效应可能与Survivin的表达下降,Saspase-3的表达上升有关.     

百里醌抑制膀胱癌细胞生长的实验研究

目的探讨百里醌对体外膀胱癌细胞株BIU-87生长的影响及其机制。方法百里醌单独作用人膀胱癌细胞株BIU-87及预处理NF-κB激活剂TPA后百里醌再作用BIU-87细胞,四甲基偶氮唑蓝(MTT)法检测细胞增殖;流式细胞术检测细胞凋亡;Western blotting检测膀胱癌细胞质中IκBα蛋白表达。结果与对照组相比较,百里醌可显著抑制体外膀胱癌细胞株BIU-87生长,并明显诱导细胞凋亡;预处理TPA可显著削弱百里醌对BIU-87细胞的增殖抑制和凋亡诱导作用;百里醌干预后膀胱癌细胞质中IκBα的表达明显上调。结论百里醌可显著抑制体外膀胱癌细胞生长,并诱导细胞凋亡,该作用可能通过促进其胞质中IκBα的表达而实现。     

中华眼镜蛇毒膜毒素诱导人膀胱癌细胞凋亡的实验研究

目的 研究中华眼镜蛇毒膜毒素12(MT-12诱导人膀胱癌细胞凋亡的作用.方法 不同浓度的MT作用于体外培养的BIU-87和E-J细胞,分别在12、24和36h用HE染色、透射电镜观察细胞的形态变化,Annexin'V-FITC/PI双染流式细胞术定量榆测凋亡率.结果 倒置显微镜和透射电镜下观察到凋亡细胞的核固缩、染色体边集、核碎裂和凋亡小体等形态学变化.MT-12浓度分别为1、2和3μg/mL时,双染法流式细胞术均检测到2株细胞凋亡率,实验组凋亡率和对照组比较有统计学意义(P<0.05).MT-12剂量和作用时间相同时,BIU-87细胞的凋亡率明显高于E-J细胞(P<0.01.结论 MT-12能诱导体外培养的BIU-87和E-J细胞凋亡,2株细胞凋亡率存在差异,可能与肿瘤细胞的生物学行为、MT-12诱导2株细胞凋亡机制的不同有关.     

As2O3诱导人膀胱癌BIU-87细胞株凋亡及其对细胞周期的影响

目的:观察三氧化二砷(As2O3)对人膀胱癌BIU-87细胞株生长抑制作用,并探讨对细胞周期及凋亡的影响.方法:CellTiter 96 Aqueous One 试剂检测BIU-87细胞株增殖活力;荧光染色观察凋亡细胞的形态学变化;流式细胞术分析细胞周期及凋亡.结果:As2O3与BIU-87细胞株生长抑制率之间有明显的浓度依赖关系 (P<0.01),IC50为28.92 μmol/L;荧光显微镜下观察到经As2O3作用的BIU-87细胞核膜消失,核染色质呈大小不等的小圆形碎片;流式细胞术结果表明30 μmol/L的As2O3处理BIU-87细胞株48 h后,G0/G1期细胞比例下降,S期细胞比例增加,凋亡率为47.37%.结论:As2O3能显著抑制BIU-87细胞株增殖;使细胞阻滞在S期,并进一步诱导细胞凋亡.     

紫杉醇诱导黑色素瘤细胞凋亡的研究

目的：探讨紫杉醇体外抑制人黑色素瘤细胞（A375细胞）增殖及诱导凋亡作用机制。方法：四甲基偶氮唑蓝（MTT）还原法测定细胞活力；光镜和电镜观察细胞形态变化；流式细胞术（FCM）检测细胞周期和凋亡率。结果：紫杉醇通过诱导细胞凋亡抑制A375细胞生长。并呈时间和剂量依赖性（P〈0．01）；当0．01-1μmol／L紫杉醇作用于培养细胞24h后即可引起猾亡小体等典型细胞凋亡形态学改变；FCM分析显示：紫杉醇在0．001μmol／L即可诱导细胞凋亡，但不影响细胞周期，在0．01μmol／L时使G0／G1期细胞明显减少，在0．1μmol／L以上时使细胞发生G2／M期阻滞。结论：紫杉醇具有抑制A375细胞生长和诱导细胞凋亡作用。不同浓度紫杉醇诱导细胞凋亡的作用机制不同。     

杨梅素诱导膀胱癌细胞株BIU-87凋亡机制研究

目的:研究杨梅素(Myricetin)诱导膀胱癌细胞株BIU-87凋亡的机制.方法:培养人膀胱癌细胞株BIU-87,加入不同浓度杨梅素干扰,在倒置显微镜下观察细胞形态学变化;并利用MTT及Hoechst 33258染色法检测杨梅素对膀胱癌细胞株BIU-87凋亡的影响;后用RT-PCR检测细胞凋亡相关基因survivin及caspase-3转录水平的改变,免疫印迹法检测survivin和caspase-3的表达情况.结果:杨梅素能诱导BIU-87细胞凋亡,并明显抑制survivin的转录和表达,同时对caspase-3有上调作用.结论:杨梅素能诱导膀胱癌细胞BIU-87凋亡,其机制与抑制survivin表达,上调caspase-3表达有关.     

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.