丹参多糖促进自噬抑制脂多糖诱导心肌细胞凋亡的实验研究

目的探讨丹参多糖(SMP)调控脂多糖(LPS)诱导的心肌细胞凋亡的作用机制。方法不同浓度(0、0.5、1、2mg/mL)SMP预处理SD大鼠心肌细胞12h后,1μg/mL LPS刺激大鼠心肌细胞24h,以LPS单独处理构建心肌细胞损伤模型,对照组加入等量生理盐水,四唑盐比色法(MTT)观察心肌细胞存活率,流式细胞仪检测心肌细胞凋亡率,免疫印迹(Western blot)检测各组心肌细胞自噬相关蛋白LC3、Beclin1及凋亡相关蛋白Bax、Caspase3表达量;SMP联合自噬抑制剂氯喹(CQ)或自噬激活剂雷帕霉素靶蛋白抑制剂(Torin1)处理LPS处理的大鼠心肌细胞24h,MTT观察细胞存活率,流式细胞仪(FACS)检测细胞凋亡率,Western blot检测各组细胞自噬相关蛋白LC3、Beclin1及凋亡相关蛋白Bax、Caspase3表达量。结果与对照组比较,LPS处理24h后,心肌细胞活力显著下降[(43.57±1.07)%比(100.03±2.12)%,P<0.01],凋亡细胞率升高[(30.34±3.13)%比(4.02±0.13)%,P<0.01],凋亡相关蛋白Bax、cleaved-Caspase3表达量增多,自噬相关蛋白Beclin1和LC3II表达量增多。与LPS组比较,1、2mg/mL SMP预处理组心肌细胞存活率显著增加[(69.72±2.15)%、(89.65±3.23)%比(43.57±1.07)%,P<0.05,P<0.01],凋亡率显著降低[(19.18±1.34)%、(12.28±1.05)%比(30.34±3.13)%,P<0.05,P<0.01],凋亡相关蛋白Bax和Cleaved-Caspase3表达量减少,自噬相关蛋白Beclin1和LC3II蛋白表达增加。与SMP处理组比较,CQ抑制细胞活力[(60.01±1.99)%比(79.42±4.23)%,P<0.05],增加细胞凋亡率[(35.63±2.45)%比(20.13±1.65)%,P<0.05],促进Bax、Cleaved-Caspase3和LC3II蛋白表达,抑制Beclin1蛋白表达;Torin1上调细胞活力[(85.63±1.08)%比(79.42±4.23)%,P<0.05],减少细胞凋亡率[(9.78±0.74)%比(20.13±1.65)%,P<0.05],促进LC3-II、Beclin1蛋白表达,抑制Bax、cleaved-Caspase3蛋白表达。结论SMP能促进自噬抑制LPS诱导的心肌细胞凋亡。     

激动核受体RXR对大鼠心肌细胞饥饿损伤中自噬和凋亡的双重调控作用

目的探讨激动维甲类X受体(RXR)对大鼠心肌细胞无糖无血清（SGD）饥饿状态下的调控作用及其机制。方法将大鼠心肌细胞株H9c2置于无糖DMEM中培养6h,建立饥饿模型。用9-顺式维甲酸（c-RA）为RXR激动剂,HX531为RXR拮抗剂,并将细胞实验按以下分组：对照组（C组）、SGD干预组（SGD组）、SGD+100nmol/Lc-RA干预组（RA组）、SGD+100nmol/Lc-RA+2.5滋mol/LHX531干预组（HX组）,MTT法检测细胞活性,流式细胞法检测细胞凋亡比例及含半胱氨酸的天冬氨酸蛋白水解酶（Caspase）-3活力,Westernblot法检测凋亡相关蛋白Bcl-2、Bax及下游活性片段Caspase-9蛋白表达,以及自噬相关蛋白LC3、Beclin1表达,透射电镜检测心肌细胞自噬小体。结果与C组比较,SGD组和HX组心肌细胞活力均下降,凋亡百分比、Bax/Bcl-2比值和Caspase-9蛋白水平均上升（均P＜0.01）;与SGD组比较,RA组心肌细胞活力均上升,凋亡百分比、Bax/Bcl-2比值和Caspase-9蛋白水平均下降（均P＜0.01）。SGD组平均荧光强度（MFI）波峰明显右移,为C组的2.3倍（P＜0.01）;而RA波峰出现明显左移,MFI值仅为C组的1.3倍,明显低于SGD组（P＜0.01）;HX组波峰与SGD组相似,MFI值显著高于C组（P＜0.01）。透射电镜下观察到C组中细胞质内细胞器完整,而SGD组和HX组自噬小体数增加,内含有被吞噬的线粒体等结构,部分呈空泡样改变,同时自噬相关蛋白LC3-II/I比值上升,Beclin1表达增加（均P＜0.01）;RA组心肌细胞自噬小体数量明显减少,LC3-II/I比值下调,Beclin1蛋白表达水平降低（均P＜0.01）。结论激动核受体RXR可以保护心肌细胞SGD状态下的损伤,其机制与对凋亡和自噬通路的双重调控作用有关。     

丹皮酚对大肠癌细胞增殖、凋亡及自噬的影响

目的 观察丹皮酚(Paeonol)对大肠癌细胞增殖、凋亡及自噬的影响,并探讨其可能的作用机制。方法 用不同浓度的丹皮酚(0、30、60、120mg/L)在体外处理大肠癌LoVo细胞,分别用CCK-8比色法、PI/Annexin V双染法、蛋白质印迹法检测LoVo细胞活力、细胞凋亡率、细胞凋亡相关蛋白(Bax和Bcl-2)及自噬相关基因蛋白(Atg5、LC3-Ⅱ和Beclin1)的表达情况。结果 通过丹皮酚处理后,CCK-8比色法显示丹皮酚以剂量、时间依赖的方式抑制LoVo细胞的增殖;PI/Annexin V双染后流式细胞仪检测显示,随着药物浓度的增加,LoVo细胞凋亡率也逐渐增加;免疫印迹结果显示,LoVo细胞凋亡相关蛋白Bax蛋白表达增加,抗凋亡相关蛋白Bcl-2蛋白的表达降低;此外,丹皮酚能上调LoVo细胞自噬相关基因Atg5、LC3-Ⅱ和Beclin1蛋白的表达。结论 丹皮酚对大肠癌LoVo细胞有明显的抗增殖和促凋亡作用,其机制可能是通过诱导细胞自噬实现的。     

黄芪甲苷通过激活自噬抑制缺氧缺糖/复氧复糖诱导的PC12细胞凋亡

目的研究黄芪甲苷通过激活细胞自噬抑制缺氧缺糖/复氧复糖诱导PC12细胞凋亡的作用。方法取对数生长期的PC12细胞,随机分为7组:正常对照组、模型组、溶媒组、黄芪甲苷组、黄芪甲苷+自噬抑制剂组、自噬抑制剂组、自噬激活剂组。除正常对照组外,其余各组剥夺氧、糖3h后再复氧、复糖12 h,并于复氧、复糖的同时给予相应药物处理。用透射电镜观察自噬小体的变化,Western blot检测LC3-II、Beclin1、半胱氨酸天冬氨酸蛋白酶3(Caspase3)的表达,流式细胞术和TUNEL染色检测细胞凋亡。结果与正常对照组相比,模型组出现典型的自噬小体,LC3-II、Beclin1表达均增多(P0.05),同时Caspase3表达、细胞凋亡率和凋亡指数也升高(P0.05)。与模型组相比,黄芪甲苷组、自噬激活剂组自噬小体数量、LC3-II和Beclin1表达均增多(P0.05),Caspase3表达、细胞凋亡率和凋亡指数均下降(P0.05);而自噬抑制剂组自噬小体数量、LC3-II和Beclin1表达减少(P0.05),Caspase3表达、细胞凋亡率和凋亡指数升高(P0.05);溶媒组与模型组之间无显著差异(P0.05)。与黄芪甲苷组相比,黄芪甲苷+自噬抑制剂组、自噬抑制剂组自噬小体数量、LC3-II和Beclin1表达均减少(P0.05),Caspase3表达、细胞凋亡率和凋亡指数均升高(P0.05)。结论黄芪甲苷可通过激活细胞自噬而抑制缺氧缺糖/复氧复糖诱导的PC12细胞凋亡,从而发挥神经保护作用。     

Bcl-2抑制剂S1诱导的卵巢癌细胞凋亡对自噬影响的实验研究

【目的】 凋亡与自噬密切相关,但二者之间的关系尚需进一步探究。本研究利用Bcl-2抑制剂S1建立卵巢癌细胞(SKOV3)凋亡模型,初步探究S1诱导的SKOV3细胞中凋亡对自噬的影响。 【方法】 培养SKOV3细胞。用MTT法检测S1对细胞生存率的影响。用流式细胞术检测细胞凋亡率。用蛋白质印迹方法检测凋亡相关蛋白Bcl-2、Bax、caspase3和自噬相关蛋白Beclin1、LC3的变化。用免疫荧光技术检测Beclin1及LC3蛋白的表达情况。在S1作用的基础上,应用广谱caspase抑制剂Z-VAD抑制caspase后,检测SKOV3细胞中自噬相关蛋白Beclin1、LC3的变化。 【结果】 S1能够降低SKOV3细胞的生存率;增加SKOV3细胞的凋亡率。与对照组(0 h)相比,S1组(2、4、8、12、24、48 h)中的Bcl-2蛋白表达持续降低,Bax及caspase3蛋白表达持续增高,呈现出明显的时间依赖性;S1组(2、4、8、12、24、48 h)中自噬的发生也呈时间依赖性,但自噬并不是持续增强的,在12 h时,自噬相关蛋白Beclin1和LC3蛋白表达水平最高,12 h后则呈下降趋势,Beclin1和LC3的荧光结果也表现出一致的现象。与S1单独作用(8、24 h)相比较,给予广谱caspase抑制剂Z-VAD,8 h结果显示 Beclin1及LC3蛋白的表达水平基本无变化,而24 h结果显示Beclin1及LC3蛋白的表达水平显著增高,同时荧光实验也显示出同样的结果。 【结论】 在SKOV3细胞中,S1通过抑制Bcl-2、促进Bax释放,诱导细胞发生凋亡,并呈时间依赖性增强。同时S1通过抑制Bcl-2,释放自噬起始基因Beclin1,诱导自噬发生。但是自噬并未始终呈时间依赖性增强,自噬水平在S1长时间作用下反而出现下降的趋势。据此推测,随着S1诱导凋亡的不断增强,大量活化的caspase可能通过裂解自噬起始基因Beclin1,从而抑制Beclin1诱导的自噬。     

桃红四物汤促进自噬抑制氧糖剥夺诱导的心肌细胞凋亡的实验研究

目的 探讨桃红四物汤(TST)对氧糖剥夺(OGD)诱导的H9C2心肌细胞损伤的保护作用。 方法 将H9C2心肌细胞分为5组(n=3)：对照组、OGD模型组、OGD +TST含药血清处理组、OGD +TST含药血清+雷帕霉素处理组、OGD +TST含药血清+氯奎处理组。四唑盐(MTT)比色法检测细胞活力;流式细胞术检测细胞凋亡;免疫印迹法(western blot)检测自噬相关蛋白(LC3、p62)和凋亡相关蛋白(Bax、Bcl-2)的表达。结果 与对照组相比,OGD模型组细胞活力显著下降(P<0.05),细胞凋亡显著增多(P<0.05),LC3II/LC3I比值和促凋亡蛋白Bax显著增多,p62和抗凋亡蛋白Bcl-2表达减少。与OGD模型组相比,OGD +TST含药血清处理组的细胞活力显著升高(P<0.05),细胞凋亡明显减少(P<0.05),LC3II/LC3I比值、Bcl-2表达增多,p62和Bax蛋白表达减少。与OGD +TST含药血清处理组相比,OGD +TST含药血清+雷帕霉素处理组的细胞活力进一步升高(P<0.05),细胞凋亡比例进一步减少(P<0.05),LC3II/LC3I比值、Bcl-2表达进一步增多,p62和Bax蛋白表达进一步减少;而氯奎处理桃红四物汤治疗组的细胞活力下降(P<0.05),凋亡细胞比例升高(P<0.05),LC3II/LC3I比值、Bcl-2表达减少,p62和Bax蛋白表达增多。结论 桃红四物汤促进氧糖剥夺H9C2心肌细胞自噬,抑制心肌细胞凋亡,对心肌缺血起保护作用。     

自噬在电离辐射诱导的小鼠精母细胞GC-2凋亡过程中的作用

目的 探讨自噬在电离辐射诱导小鼠精母细胞GC-2凋亡过程中的作用.方法 将小鼠精母细胞GC-2分为空白对照组和不同剂量(2、4和8 Gy)60Co辐射处理组.采用原位末端转移酶标记法(TUNEL法)和流式细胞术检测细胞凋亡情况,通过蛋白质印迹实验观察自噬相关蛋白LC3(LC3-Ⅰ、LC3-Ⅱ)和Beclin1表达水平的改变,使用荧光显微镜观察C,C-2细胞内自噬小体的变化.用5mmol/L自噬抑制剂3-甲基腺嘌呤(3-MA)处理GC-2细胞2h再给予60Co辐射处理,观察自噬抑制剂联合电离辐射对细胞凋亡率的影响以及自噬相关蛋白表达水平的改变.结果 与空白对照组相比,GC-2细胞经60Co辐射处理后细胞凋亡率升高(P＜0.05),荧光显微镜下可见细胞自噬体明显增多,自噬相关蛋白LC3-Ⅱ以及Beclin1蛋白表达水平增加(P＜0.05).预先用5mmol/L 3-MA处理GC-2细胞2h再给予60Co辐射处理,自噬相关蛋白LC3-Ⅱ以及Beclin1蛋白表达水平与未经3-MA处理组相比降低(P＜0.05),细胞凋亡率升高(P＜0.05).结论 电离辐射可以诱导精母细胞发生自噬,抑制细胞自噬后可增强电离辐射对精母细胞的杀伤作用.     

凉血化瘀方对肝细胞损伤中自噬与凋亡相关蛋白的影响

目的 研究凉血化瘀方对肿瘤坏死因子α（TNF-α）联合D-氨基半乳糖(D-GalN)所致肝细胞损伤中自噬和凋亡相关途径的影响,并探讨其分子调控机制。方法 体外培养人正常肝LO2细胞,用TNF-α（100 ng/mL）和D-GalN（44 μg/mL）诱导肝细胞损伤模型,荧光显微镜观察细胞的形态学变化;MTT法检测细胞活力;Western blot检测凋亡和自噬标志蛋白caspase-3、Beclin1、LC3Ⅰ、LC3Ⅱ,凋亡和自噬调节蛋白Bax、Bcl-2和mTOR信号通路调控途径mTOR蛋白表达水平的变化。结果 凉血化瘀方能改善肝细胞病理损伤,并表现有剂量依赖性;治疗组细胞凋亡和自噬标志蛋白caspase-3、Beclin1、LC3Ⅱ表达下调,凋亡和自噬调控蛋白中负相关的Bcl-2表达上调、正相关的Bax表达下调,并呈现一定的量效关系,同时mTOR蛋白的表达也体现了剂量相关的下调作用。结论 凉血化瘀方对TNF-α+D-GalN所致肝细胞损伤的保护作用,与抑制细胞自噬和凋亡相关,其机制主要是参与自噬和凋亡相关途径中蛋白的表达调控。     

茶多酚对卵巢癌SKOV3细胞自噬水平的影响及相关机制研究

目的研究茶多酚(EGCG)对人卵巢癌SKOV3细胞自噬水平的影响及相关机制。方法用EGCG处理SKOV3细胞,Western blot检测自噬相关蛋白LC3-II及蛋白激酶B(PKB)信号通路相应蛋白表达变化。结果 EGCG处理SKOV3细胞后,自噬相关蛋白LC3-II表达上调,并呈一定的时间浓度依赖性。EGCG处理SKOV3细胞后,AKT的磷酸化水平下调,AKT激活剂胰岛素样生长因子1(IGF-1)预处理后,自噬相关蛋白LC3-II高表达被抑制。结论 EGCG通过AKT介导的信号通路诱导SKOV3细胞自噬水平升高。     

自噬调控对小鼠精原细胞缺氧/复氧损伤的影响研究

目的 研究自噬调控对小鼠精原细胞缺氧/复氧损伤(H/RI)的影响,探讨自噬对小鼠睾丸组织缺血再灌注损伤(IRI)的影响。方法 以小鼠精原细胞系GC1 spg为研究对象构建H/RI模型,雷帕霉素(RAPA)和3-甲基腺嘌呤(3-MA)分别作为自噬激动剂和抑制剂,设置对照组、模型组(H/RI)、自噬激动剂干预组(H/RI+自噬激动剂)、自噬抑制剂干预组(H/RI+自噬抑制剂)。用噻唑蓝(MTT)法检测各组细胞增殖能力,流式细胞术检测各组细胞活性氧(ROS)释放水平和凋亡水平,实时荧光定量PCR(qPCR)检测各组细胞自噬相关基因Beclin1和凋亡相关基因Bcl-2、Bax mRNA的表达水平,Western blot检测各组细胞自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ、Beclin1、p62和凋亡相关蛋白Bcl-2、Bax、caspase-3的表达水平。结果 与对照组比较,模型组增殖能力、Beclin1和Bcl-2 mRNA表达水平明显下降(P<0.01),p62和Bcl-2蛋白表达水平明显下降(P<0.01),ROS水平、细胞凋亡率、Bax mRNA相对表达水平明显上升(P<...     

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.