去甲斑蝥素体外抑制NCI-H446细胞迁移、增殖及促进凋亡

目的 研究去甲斑蝥素(Norcantharidin,NCTD)体外抑制人小细胞肺癌H446细胞迁移、增殖和诱导其凋亡作用.方法 以不同浓度NCTD处理H446细胞24h后,采用细胞划痕试验检测NCTD对H446细胞侧向迁移能力的影响;采用细胞计数检测对体外培养的H446细胞的抑制增殖作用;选用不同浓度的NCTD作用H446细胞48 h后,以Hoechst33258染色观察H446细胞形态的变化,研究NCTD对人小细胞肺癌H446细胞凋亡的诱导作用.结果 经较高浓度的NCTD干预24h后,H446细胞的侧向迁移速度明显减慢(P＜0.01).细胞计数结果显示NCTD作用于H446细胞后其细胞群体倍增时间延长.NCTD作用H446细胞48 h后,部分凋亡细胞出现染色质凝集、核固缩、核碎裂等形态学改变,细胞凋亡率增加(P＜0.01).结论 NCTD能明显影响人小细胞肺癌H446细胞迁移、抑制其增殖并诱导其发生凋亡.     

姜黄素抑制刺猬信号诱导小细胞肺癌细胞凋亡

目的：探讨姜黄素对小细胞肺癌NCI- H446细胞的作用及可能机制。方法不同浓度（5、10、15μmol/L）的姜黄素作用于小细胞肺癌NCI- H446细胞后,采用MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot技术检测索尼克刺猬信号（Shh）、脑胶质瘤相关癌基因1（Gli1）的表达水平。结果不同浓度的姜黄素与小细胞肺癌NCI- H446细胞共培养后,浓度为15μmol/L的姜黄素能明显抑制NCI- H446细胞的增殖。与其余各组相比,差异有统计学意义（P＜0.01）。小细胞肺癌NCI- H446细胞经姜黄素处理后,姜黄素15μmol/L与其余各组比较,细胞凋亡率的差异有统计学意义（P＜0.01）。经姜黄素（15μmol/L）处理的NCI- H446细胞,Shh和Gli1表达均明显降低,与对照组相比,差异均有统计学意义（P＜0.01）。结论姜黄素能通过抑制刺猬信号通路,抑制小细胞肺癌细胞的增殖,诱导其凋亡。     

吴茱萸碱抑制PI3K/AKT通路诱导小细胞肺癌H1688和H446细胞凋亡

目的 探讨PI3 K/AKT通路在吴茱萸碱(evodiamine,EVO)诱导小细胞肺癌H1688和H446细胞凋亡中的作用.方法 分别以不同浓度(1、5、20 μmol/L)和不同作用时间(3、6、12、24、48 h)的EVO处理H1688和H446细胞,Western blot检测p-AKT的表达水平.以EVO分别作用于经过PI3 K/AKT通路激活剂IGF-1或抑制剂LY294002预处理后的H1688和H446细胞,Western blot检测p-AKT蛋白的表达.将EVO作用于IGF-1预处理的H1688细胞后,Annexin-V/PI染色检测细胞凋亡率,Western blot检测Bcl-2、Bax、Caspase-3蛋白的表达水平.以不同浓度的EVO处理H1688细胞,Western blot检测t-AKT、PTEN蛋白表达,RT-PCR检测AKT mRNA表达.结果 与对照组相比,不同浓度和不同作用时间的EVO处理后H1688和H446细胞中p-AKT表达水平下降(P＜0.05).而IGF-1可减弱EVO下调H1688和H446细胞p-AKT的作用(P＜0.05),并降低EVO在H1688中的增殖抑制和凋亡诱导作用.EVO与LY294002下调p-AKT蛋白的效果差异无统计学意义(P＞0.05).EVO对H1688细胞t-AKT蛋白和AKT mRNA的表达无影响,EVO可明显上调PTEN的表达(P＜0.05).结论 抑制PI3 K/AKT通路活性是EVO诱导H1688和H446细胞凋亡的重要机制之一.     

FoxM1通过Wnt/β-catenin信号通路对鼻咽癌细胞5-8F增殖、迁移的影响

目的 探讨FoxM1对鼻咽癌细胞5-8F恶性生物学行为的影响及其可能机制.方法 将化学合成的靶向FoxM1的siRNA转染鼻咽癌细胞5-8F(FoxM1-siRNA组),并设立空白对照组、阴性对照组,采用RT-PCR及Western blot检测FoxM1表达变化.采用MTT法、FCM法、Transwell法分别检测下调FoxM1表达对细胞增殖、周期、凋亡、迁移的影响.Western blot检测下调FoxM1表达后β-catenin、C-Myc、Cyclin D1、MMP-2、MMP-9蛋白表达变化.结果 转染FoxM1-siRNA的5-8F细胞FoxM1 mRNA和蛋白表达水平降低(P＜0.05).下调FoxM1表达可抑制5-8F细胞增殖(P＜0.05),FoxM1-siRNA组的凋亡率为(22.68±0.67)％,较阴性对照组[(1 1.74±1.36)％]和空白对照组[(10.94±1.52)％]显著增加(P＜0.05),细胞周期阻滞于G0/G1期.FoxM1-siRNA组细胞穿膜数(100.00±10.97)明显低于阴性对照组(246.00±6.66)和空白对照组(233.70±12.41),细胞迁移能力降低(P＜0.05).下调FoxM1的表达可抑制β-catenin细胞核表达,抑制C-Myc、Cyclin D1、MMP-2、MMP-9总蛋白表达(P＜0.001).结论 FoxM1表达下调抑制鼻咽癌细胞5-8F的增殖,诱导细胞凋亡,降低细胞迁移能力,使细胞周期阻滞于G0/G1期.其机制可能是通过抑制Wnt通路关键蛋白β-catenin细胞核表达从而抑制Wnt 通路活性实现的.     

表没食子儿茶素没食子酸酯增强低剂量顺铂对小细胞肺癌H446细胞株的细胞毒作用

目的探讨表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]联合顺铂(Cisplatin,DDP)对小细胞肺癌细胞株H446的细胞毒作用及其机制。方法采用MTT法检测EGCG及DDP影响H446细胞增殖的量效和时效关系。采用流式细胞仪检测不同浓度EGCG、DDP及二者联合作用于H446细胞对细胞周期及凋亡的影响。结果 EGCG和DDP均可抑制H446细胞增殖,该增殖抑制作用呈浓度及时间依赖模式。不同浓度的EGCG(80、160μmol/L)与DDP(1.0μg/ml)联合应用时,可使H446细胞阻滞于G2/M期。EGCG可诱导H446细胞凋亡,并能增强DDP诱导H446细胞凋亡的作用。结论 EGCG能够抑制H446细胞增殖,诱导H446细胞凋亡,增强DDP诱导H446细胞凋亡的作用,其机制可能与G2/M期阻滞有关。     

(-)-epigallocatechin-3-gallate enhances low-dose cisplatin cytotoxicity to human small cell lung cancer cell line H446

目的 探讨表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]联合顺铂(Cisplatin,DDP)对小细胞肺癌细胞株H446的细胞毒作用及其机制.方法 采用MTT法检测EGCG及DDP影响H446细胞增殖的量效和时效关系.采用流式细胞仪检测不同浓度EGCG、DDP及二者联合作用于H446细胞对细胞周期及凋亡的影响.结果 EGCG和DDP均可抑制H446细胞增殖,该增殖抑制作用呈浓度及时间依赖模式.不同浓度的EGCG(80、160 μmol/L)与DDP(1.0μg/nl)联合应用时,可使H446细胞阻滞于G2/M期.EGCG可诱导H446细胞凋亡,并能增强DDP诱导H446细胞凋亡的作用.结论 EGCG能够抑制H446细胞增殖,诱导H446细胞凋亡,增强DDP诱导H446细胞凋亡的作用,其机制可能与G2/M期阻滞有关.     

多西环素对体外人小细胞肺癌H446细胞增殖的影响及其机制

目的 探讨多西环素对体外人小细胞肺癌H446细胞增殖的影响及其可能的作用机制.方法 不同浓度的多西环素作用于体外培养的人小细胞肺癌H446细胞24、48、72或96 h,采用CCK-8法检测多西环素对H446细胞增殖的影响;采用Bliss法计算半数抑制浓度(IC50);采用TUNEL法检测多西环素对H446细胞凋亡的影响;采用Western印迹法检测多西环素对Bcl-2、Bax等凋亡相关蛋白表达的影响;采用实时定量-PCR法检测多西环素对Bcl-2、Bax等凋亡相关基因mRNA表达的影响.结果 多西环素可浓度-时间依赖性地抑制人小细胞肺癌H446细胞的体外增殖,作用24、48、72和96 h对应的IC50值分别为24.10、10.98、8.20和8.03 mg/L;多西环素作用后H446细胞的凋亡指数显著增加(P<0.05);Bax的蛋白及mRNA表达水平上调,Bcl-2的蛋白及mRNA表达水平下调,差异具有统计学意义(P<0.05).结论 多西环素可通过上调促凋亡相关蛋白Bax的表达,下调抑凋亡相关蛋白Bcl-2的表达,诱导人小细胞肺癌H446细胞凋亡,进而抑制人小细胞肺癌H446细胞的体外增殖,是一种有开发前景的小细胞肺癌治疗药物.     

葛根素对人小细胞肺癌H446细胞周期和相关周期蛋白表达的影响

目的观察葛根素诱导人小细胞肺癌H446细胞周期的阻滞作用,进一步探讨其作用机制。方法采用MTT法测定葛根素对H446细胞生长的影响;采用流式细胞术检测处理后H446细胞周期分布和周期蛋白cyclinD1、CDK4和P27表达水平。结果MTT法表明葛根素对H446细胞生长有较强的抑制作用;流式细胞仪检测结果发现细胞周期阻滞在G0/G1期,量效关系良好,并检测到细胞周期蛋白cyclin D1、CDK4表达减弱,P27表达增强。结论葛根素可通过诱导细胞周期阻滞而发挥体外抗肺癌作用,其机制可能涉及细胞周期相关蛋白cyclin D1、CDK4和P27表达的调控。     

丹参酮ⅡA诱导人小细胞肺癌细胞凋亡及其分子机制

目的 了解丹参酮ⅡA对人小细胞肺癌细胞（H446细胞）的凋亡诱导作用,并探讨其可能的分子机制。方法 采用MTT法检测不同浓度丹参酮ⅡA对人小细胞肺癌H446细胞增殖的影响结果;Hoechst 33258法检测不同浓度丹参酮ⅡA对人小细胞肺癌H446细胞凋亡的影响;qPCR法检测不同浓度丹参酮ⅡA对人小细胞肺癌H446细胞凋亡基因的影响;Westen blot法检测不同浓度丹参酮ⅡA对人小细胞肺癌H446细胞凋亡的影响。结果 不同药物浓度的丹参酮ⅡA （0.3、0.6、1.25、2.5、5和10μg/ml）抑制人小细胞肺癌H446细胞增殖且呈浓度和时间依赖性;随着药物浓度的提高,人小细胞肺癌H446细胞凋亡小体的形成数量增加,高浓度组差异有统计学意义（P<0.05）;高浓度组丹参酮ⅡA促进凋亡基因（Bax,caspase-9、caspase-3）的表达,抑制凋亡抑制基因（Akt,Bcl-2）的表达且差异有统计学意义（P<0.05）;高浓度组丹参酮ⅡA促进凋亡蛋白（Bax,caspase-9,caspase-3）的表达,抑制凋亡抑制蛋白（Akt,Bcl-2）的表达且差异有统计学意义（P<0.05）。结论 丹参酮ⅡA可能是通过Akt-Bax/bcl-2-caspase9-caspase3信号通路诱导人小细胞肺癌H446细胞的凋亡。     

抑制自噬增强门冬酰胺酶对小细胞肺癌细胞H1688和H446的抗癌作用

目的 研究门冬酰胺酶(asparaginase,AN)对小细胞肺癌H1688和H446细胞的抗癌作用,探讨自噬在AN抗癌过程中的功能.方法 采用MTT法及台盼蓝染色法检测AN单独或联用自噬抑制剂氯喹(chloroquine,CQ)对H1688和H446细胞的生长抑制作用;免疫荧光法观察自噬标记物LC3表达,示踪自噬体形成;Western blot检测自噬相关蛋白LC3及Akt/mTOR信号通路的表达.结果 AN呈浓度依赖性抑制H1688和H446细胞增殖并促进其死亡(P＜0.05);AN处理组可以显著增加H1688和H446细胞自噬小体数量并诱导LC3-Ⅱ表达;自噬抑制剂CQ提高AN对H1688和H446细胞杀伤作用(P＜0.05);AN作用H1688细胞后p-Akt、p-mTOR、p-70S6K蛋白表达降低.结论 AN对小细胞肺癌H1688和H446细胞具有抑制作用并诱导细胞保护性自噬,阻断自噬可以增强AN对小细胞肺癌H1688和H446细胞的抗癌疗效.     

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.