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。     

As2O3对人卵巢癌裸鼠皮下移植瘤抑制作用及机制

目的 探讨不同剂量的三氧化二砷(As2O3)对人卵巢癌细胞株SKOV3细胞裸鼠皮下移植瘤生长的抑制作用及其机制.方法 将4×106个SKOV3细胞接种于裸鼠皮下,建立人卵巢癌裸鼠皮下移植瘤模型,随机分为As2O3高、中、低剂量组及生理盐水组(空白对照)、卡铂(CBP)组(阳性对照),连续给药10 d,停药后24 h处死裸鼠,计算移植瘤的瘤质量、抑瘤率及caspase-3基因表达水平的变化,并检测用药后裸鼠的肝、肾功能及血常规.结果 As2O3对移植瘤生长的抑制作用具有剂量依赖性,各剂量As2O3组及CBP组瘤质量与生理盐水组比较均有统计学意义(F=17.931,P<0.05);As2O3作用后caspase-3基因的表达较空白对照组升高,差异有统计学意义(F=31.821,q=2.89～3.84,P<0.05).同时,As2O3各组与CBP组相比,未表现出明显的肝、肾及血液学毒性.结论 As2O3对人卵巢癌移植瘤的生长具有明显的抑制作用,其机制可能与上调caspase-3基因的表达及诱导细胞凋亡有关.     

三氧化二砷和顺铂对人卵巢癌细胞株3AO体外杀伤效应的对比研究

目的 :比较As2 O3 和顺铂 (cDDP)对卵巢癌细胞株的生长抑制作用和凋亡诱导作用。方法 :采用MTT测定法和流式细胞分析法 (FCM) ,检测As2 O3 对体外培养的人卵巢癌细胞株 3AO细胞的生长抑制率和诱导凋亡率及对细胞周期的影响 ,并与cDDP进行比较。采用吖啶橙染色 ,观察As2 O3 作用后 ,凋亡细胞的形态学变化。结果 :As2 O3 可以有效地抑制 3AO细胞生长并诱导其凋亡 ,具有量 效性及时 效性 ,与cDDP相比 ,差异有统计学意义 (P <0 .0 1)。As2 O3 作用后 ,S期细胞通过受阻 ,形态学观察发现细胞呈典型的凋亡细胞特征。结论 :与cDDP相比 ,As2 O3 对卵巢癌细胞株 3AO细胞具有更有效的生长抑制作用及诱导凋亡作用 ,并使S期细胞阻滞。     

As2O3对人宫颈癌裸鼠皮下移植瘤的抗肿瘤作用

目的 探讨不同剂量三氧化二砷(As2O3)对人宫颈癌裸鼠皮下移植瘤的抗肿瘤作用及机制.方法 将4×106个Hela细胞接种于裸鼠皮下,建立人宫颈癌裸鼠皮下移植瘤模型,随机分为As2O3低剂量(1.0 mg/kg)、中剂量(2.5 mg/kg)、高剂量(5.0 mg/kg)组及生理盐水空白对照组、顺铂阳性对照组,腹腔连续给药10 d,停药后24 h处死裸鼠留取标本计算抑瘤率,流式细胞仪检测细胞凋亡率, RT-PCR方法检测移植瘤组织caspase-3基因的相对表达量,免疫组化SP法检测磷酸化细胞外信号调节激酶(pERK)的表达.结果 各剂量As2O3组及顺铂组瘤质量与生理盐水组相比均有统计学意义(F=39.14,P<0.05);As2O3高、中、低剂量组流式细胞仪检测时均出现典型凋亡峰,凋亡率分别为17.89%、17.19%、18.52%;As2O3作用后caspase-3基因的表达较生理盐水组明显升高,差异均有统计学意义(F=8.76,q=3.06～5.81,P＜0.05);As2O3各剂量组pERK的表达较生理盐水组降低.结论 As2O3 对人宫颈癌移植瘤的生长具有明显的抑制作用,其机制与上调caspase-3基因的表达诱导细胞凋亡,下调pERK的表达与抑制MAPK信号传导通路有关.     

人卵巢癌顺铂耐药细胞裸鼠腹腔转移模型的建立

目的　探讨建立人卵巢癌顺铂耐药细胞裸鼠腹腔转移模型的方法。方法　人卵巢癌细胞株 3AO和顺铂耐药细胞株 3AO/cDDP细胞体外培养。 4× 10 6个 3AO、3AO/cDDP分别接种于裸鼠腹腔 96h后 ,各随机分为 2组 ,分别腹腔内注射生理盐水和顺铂 (cDDP) ,观察各组裸鼠的致瘤率、生存期和生存延长率。采用流式细胞技术观察转移瘤LRP、MRP基因的表达情况。转移瘤瘤体行病理学检查。结果　 3AO、3AO/cDDP细胞的裸鼠致瘤率差异无显著性 ,P >0 .0 5 ;cDDP能提高 3AO荷瘤鼠的生存期及生存延长率 ,与 3AO/cDDP荷鼠相比 ,差异有显著性 ,P <0 .0 5 ;3AO/cDDP转移瘤的LRP、MRP基因的表达水平显著高于 3AO转移瘤 ,P <0 .0 5 ;病理学检查提示裸鼠腹腔转移瘤细胞具人恶性特征。结论　该方法建立裸鼠卵巢癌腹腔转移模型具有成瘤率高的特点 ,并能保持顺铂耐药特征。     

喷他脒对卵巢癌Skov-3细胞及其裸鼠移植瘤PRL-3的抑制作用

目的 探讨喷他脒对卵巢癌Skov-3细胞及其裸鼠移植瘤PRL-3的抑制作用.方法 (1)M TT法检测不同浓度喷他脒对Skov-3细胞的生长抑制作用;(2)RT-PCR法检测不同浓度喷他脒处理后的Skov-3细胞PRL-3的表达;(3)建立人卵巢癌Skov-3 细胞裸鼠转移瘤模型,按喷他脒给药剂量分4组,记录裸鼠皮下移植瘤的生长情况,绘制肿瘤生长曲线.采用RT-PCR方法检测裸鼠移植肿瘤的PRL-3表达情况.结果 喷他脒对卵巢癌Skov-3细胞体外生长有抑制作用,呈浓度依赖性(P＜0.05);喷他脒中高剂量组予5次给药后裸鼠移植瘤生长速度均与对照组有差异(P＜0.05);中、高剂量喷他脒抑制人Skov-3卵巢癌细胞裸鼠皮下移植瘤的PRL-3表达,且随剂量增加而抑制作用更明显(P＜0.05).结论 喷他脒对卵巢癌Skov-3细胞体外生长及裸鼠Skov-3皮下移植瘤均有有抑制作用;喷他脒可抑制Skov-3卵巢癌细胞及裸鼠Skov-3皮下移植瘤PRL-3的表达.     

RNAi沉默STAT3基因对人卵巢癌SKOV3细胞裸鼠移植瘤生长的抑制作用

目的：研究pSilencer1.0-U6-siRNA-STAT3重组质粒对人卵巢癌SKOV3细胞裸鼠皮下移植瘤生长的抑制作用,阐明RNA干涉技术在卵巢癌生物治疗领域中的作用。方法：应用人卵巢癌细胞系SKOV3对15例裸鼠建立人卵巢癌皮下移植瘤模型,随机分为3组：对照组（生理盐水）、空质粒对照组及重组质粒组（pSilencer1.0-U6-siRNA-STAT3）。应用电子穿孔仪将质粒转入裸鼠移植瘤内,观察重组质粒注射后第7、14、21和28天各组裸鼠皮下移植瘤体积变化。利用Western blotting检测重组质粒对STAT3蛋白表达的影响,采用HE及TUNEL染色方法观察肿瘤组织形态变化及细胞凋亡情况。结果：重组质粒瘤内注射对SKOV3裸鼠皮下移植瘤的生长有明显的抑制作用,与2个对照组比较,重组质粒组裸鼠移植瘤生长速度明显减慢（P<0.01）。重组质粒组瘤体内STAT3及CyclinD1、VEGF、survivin、c-myc蛋白表达明显低于2个对照组（P<0.01）。HE染色显示,重组质粒组肿瘤细胞出现大片细胞坏死,2个对照组肿瘤细胞以正常形态居多。TUNEL染色显示,重组质粒组癌组织有大量细胞凋亡,2个对照组几乎均为TUNEL阴性反应细胞。结论：pSilencer1.0-U6-siRNA-STAT3重组质粒能明显抑制人卵巢癌细胞SKOV3裸鼠皮下移植瘤的生长。     

三氧化二砷抑制食管癌细胞EC-9706体内增殖及机制研究

目的:研究三氧化二砷(As2O3)对食管癌细胞系EC-9706的体内增殖抑制作用及可能机制。方法:以不同剂量的As2O3作用于裸鼠移植瘤模型,每周测量裸鼠移植瘤的大小,绘制肿瘤生长曲线;采用Western blot法检测Pim-3和B细胞淋巴瘤/白血病-2(Bcl-2)蛋白的表达变化。结果:不同剂量的As2O3均能明显抑制裸鼠移植瘤的生长,以4 mg/kg As2O3组的抑制作用最强,肿瘤重量抑制率和肿瘤体积抑制率分别为81.13%和81.48%。Pim-3和Bcl-2蛋白的表达受到抑制。结论:As2O3可能通过抑制Pim-3使Bcl-2表达降低,促进肿瘤细胞凋亡。     

中华眼镜蛇毒抗瘤多肽对人卵巢癌HO-8910细胞的增殖抑制和Fas/FasL表达的影响

目的 探讨中华眼镜蛇毒抗瘤多肽对人卵巢癌HO-8910细胞株的抑制作用及Fas/FasL表达的影响.方法 用MTT比色法检测不同浓度中华眼镜蛇毒抗瘤多肽对HO-8910细胞的增殖抑制,RT-PCR和免疫组织化学观察原癌基因Fas/FasL表达的变化.结果 中华眼镜蛇毒抗瘤多肽对HO-8910细胞增殖抑制作用成剂量依赖性,RT-PCR和免疫组化结果显示,随着中华眼镜蛇毒抗瘤多肽作用剂量的增加,Fas/FasL基因的表达无明显变化.结论 中华眼镜蛇毒抗瘤多肽对人卵巢癌HO-8910细胞系的增殖具有明显的抑制作用,并对原癌基因Fas/FasL的表达无抑制作用.     

重组凋亡素对人卵巢癌裸鼠移植瘤生长抑制作用的影响

目的 观察凋亡素(VP3)基因真核表达质粒对人卵巢癌裸鼠异种移植瘤生长的抑制作用.方法 建立人卵巢癌(COC1)细胞裸鼠异种移植瘤模型.实验裸鼠分为四组:高剂量实验组(pcDNA-VP3质粒DNA,瘤内注射,100μg/只);低剂量实验组(pcDNA-VP3质粒DNA,瘤内注射,50μg/只);空载体对照组[pcDNA3.1(+)质粒DNA,瘤内注射,100μg/只];脂质体对照组(脂质体,瘤内注射,0.2mL/只).每3d测量皮下移植瘤长、宽径,按标准公式V=L×W2X0.52计算移植瘤体积.12天后处死裸鼠,称取瘤重.使用TUNEL法观察凋亡素对皮下移植瘤细胞凋亡程度的影响.结果 pcDNA9·vP3高剂量组和低剂量组卵巢癌裸鼠皮下移植瘤生长明显受抑制.高剂量组和低剂量组肿瘤生长抑制率分别为76%和52%;瘤重抑制率分别为71.9%和49.5%(P<0.05).TUNEL实验结果表明凋亡素在体内以凋亡方式诱导肿瘤细胞死亡.结论 重组凋亡素基因真核表达质粒具有抑制人卵巢癌裸鼠异种移植瘤生长作用.凋亡素在瘤组织中以诱导凋亡方式抑制卵巢癌裸鼠异种移植瘤生长.     

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.