阿托伐他汀对血管内皮细胞增殖过程中FKBP12基因表达的影响

目的探讨不同浓度阿托伐他汀对血管内皮细胞（VEC）增殖过程中FK506结合蛋白12（FKBP12）表达的影响。方法培养大鼠VEC并传代,检测Ⅷ因子相关抗原。观察其不同浓度阿托伐他汀作用下的VEC增殖情况和不同时期FKBP12 mRNA水平的表达。结果阿托伐他汀抑制大鼠血管内皮细胞的增殖过程,抑制血管内皮细胞增殖过程中FKBP12的表达,该作用呈时间依赖性和剂量依赖性;在阿托伐他汀作用下,大鼠血管内皮细胞数随浓度的增加和作用时间的延长呈相对减少趋势。结论阿托伐他汀抑制血管内皮细胞的增殖过程,抑制血管内皮细胞增殖过程中FKBP12的表达,且呈时间-剂量依赖性。     

阿托伐他汀对血管内皮细胞增殖过程中FKBP12mRNA和细胞上清液ET-1的影响

目的探讨不同浓度阿托伐他汀对血管内皮细胞（VEC）增殖过程中FK506结合蛋白12（FKBP12）表达和细胞上清液ET-1的影响。方法培养大鼠VEC并传代,观察其不同浓度阿托伐他汀作用下的不同时期FKBP12mRNA水平的表达和细胞上清液ET-1的变化。结果阿托伐他汀抑制血管内皮细胞增殖过程中FK—BP12的表达,抑制血管内皮细胞合成和分泌ET-1;血管内皮细胞FKBP12表达和ET-1分泌基本一致,呈正相关,阿托伐他汀对它们的抑制作用且呈时间依赖性和剂量依赖性。结论阿托伐他汀通过降低FKBP12表达抑制血管内皮细胞增殖并对ET-1合成和分泌起负性调节作用。     

胰岛素对VEC增殖过程中FKBP12基因表达的影响

目的　通过不同浓度胰岛素对血管内皮细胞 (vascularendothelialcell ,VEC)增殖过程中胰岛素受体 (insulinre ceptor ,InsR)和FK5 0 6结合蛋白 12 (FK5 0 6bindingprotein 12 ,FKBP12 )及其基因表达等的影响 ,初步探讨VEC的胰岛素抵抗。方法　在内皮细胞生长因子 (endothelialcellgrowthfactor ,ECGF)作用下 ,培养大鼠VEC并传代。检测VEC在增殖过程中不同浓度胰岛素对VEC的InsR和FKBP12及其基因表达、生长曲线和细胞上清液一氧化氮 (nitrogenmonoxide ,NO)含量等的影响。结果　与对照组相比 ,1U/ml胰岛素组VEC的InsR和FKBP12mRNA表达降低 ;生长曲线向右下偏移且对数生长期延迟 ;细胞上清液NO含量下降。结论　VEC是胰岛素敏感细胞 ;当培养液中胰岛素浓度为 1U/ml时 ,经 16h的培养后 ,可使VEC对生理浓度胰岛素的反应性降低 ,该胰岛素抵抗状态至少维持 2 4h。FKBP12可能是与VEC胰岛素抵抗相关的重要信息分子。     

人血管内皮细胞在体外连续增殖传代的研究

作者用胶原酶消化法自人脐带静脉分离获取内皮细胞,由牛下丘脑提取内皮细胞生长因子(ECGF),以改进的方法对内皮细胞进行培养,建立起在体外连续增殖传代的人血管内皮细胞(HEC)的培养模型。并就ECGF和人纤维粘连蛋白(HFN)对HEC生长的生物学效应进行观察和比较。结果表明,①HEC在低密度(5×10~3/cm~2)情况下,即对ECGF(100～300mg/L)有良好的增殖反应,可在体外连续增殖传至10代;②HFN对HEC增殖无直接促进作用,但预先用其涂铺培养瓶底面,有利于细胞贴壁:③ECGF和HFN不能替代小牛血清。此培养体系的建立,将为内皮细胞的生理和病理研究提供便利条件。     

FKBP12蛋白RNA干扰与回复表达细胞系的建立

目的建立FKBP12蛋白(FK506 binding-protein 12,基因名FKBP1A)RNA干扰和回复表达的人肺癌细胞系A549,并初步探索FKBP12的功能。方法设计3条核心序列位于FKBP1A 3’UTR区域的siRNA,它们仅靶向内源性的FKBP12。利用p LKO.1-puro载体包装相应的shRNA慢病毒感染A549,得到FKBP12表达调低的细胞系。使用带neo基因筛选标记的pc DNA3.1载体回复表达FKBP12,并筛选得到不受shRNA干扰的回复表达细胞系。用实时荧光定量PCR和Western blotting检测调低和回复表达效果,并初步分析细胞系表型。结果 FKBP12在mRNA和蛋白质水平上的表达均被抑制,干扰效率在75%以上(P<0.01)。FKBP12不影响A549的细胞形态和增殖效率,而下调m TORC1(mammalian target of Rapamycin complex 1)下游转录调控相关蛋白4E-BP1(e IF4E-binding protein 1)的表达。结论成功构建了FKBP12蛋白敲低和回复表达的A549细胞系,为后续探究FKBP12的功能奠定了基础。     

碘离子对血管内皮细胞表达bcl-2/bax蛋白的影响及布加综合征隔膜形成机制

目的探讨碘离子对血管内皮细胞(vascular endothelial cell,VEC)表达bcl-2/bax蛋白的影响。方法在不同浓度的碘离子环境中体外培养VEC,采用蛋白质印迹技术(Western Blot)检测VEC中bcl-2/bax蛋白表达情况。结果不同浓度的碘离子作用于VEC后,凋亡蛋白bcl-2及bax的表达量均未出现上调(P>0.05)。结论碘离子促进VEC增殖,并不是通过调控bcl-2/bax蛋白的表达量而实现的。     

FKBP1B在乳腺癌中的表达及其临床意义

目的 明确FK506结合蛋白1B(FK506 binding proteins 1B,FKBP1B)在乳腺癌中的表达情况及其对乳腺癌细胞的增殖和迁移能力的影响及机制。方法 利用商业化的乳腺癌组织芯片进行免疫组织化学检测FKBP1B的表达,共包含乳腺癌组织及乳腺癌旁组织的石蜡标本72例,并分为癌旁组织3例,肿瘤组织69例。应用实时荧光定量PCR(qRT-PCR)和Western blot法检测乳腺癌细胞系MCF-7和MDA-MB-231中FKBP1B的表达情况。在MCF-7细胞中过表达FKBP1B后,分别用CCK-8和Transwell检测FKBP1B对细胞增殖和迁移能力的影响。并用Western blot法检测FKBP1B对热休克蛋白90α(Hsp90α)的表达和分泌的影响。结果 FKBP1B在乳腺癌肿瘤组织中的含量显著高于正常组织,且FKBP1B的表达与TNM分期之间具有显著相关性。RT-PCR和Western blot法表明,FKBP1B在MDA-MB-231中的含量显著高于MCF-7。过表达FKBP1B后,MCF-7的细胞增殖和迁移能力较对照组明显增强。FKBP1B的表达与Hsp90α的分泌呈正相关,过表达FKBP1B能够促进Hsp90α的分泌。结论 FKBP1B在乳腺癌中表达增高且与疾病恶性进展相关,过表达FKBP1B能够促进Hsp90α的分泌以及乳腺癌细胞的增殖和迁移能力,其可作为乳腺癌诊断和治疗的新的潜在分子靶点。     

反义MMP-2基因影响人增殖期血管瘤内皮细胞MMP-2表达的体外研究

目的 研究反义基质金属蛋白酶-2基因(aMMP-2)对体外堵养的增殖期血管瘤内皮细胞(VEC)MMP-2的影响.方法 实验分为M199组(无血清M199培养VEC)、Ad-GFP组(用Ad-GFP感染VEC) Ad-aMMP-2组(用Ad-aMMP-2感染VEC),感染复数(MOI)均为100.分别用RT-PCR、Western Blotting和明胶酶试验从基因水平和蛋白水平上检测各组VEC MMP-2基因的表达.结果 与M199组和Ad-GFP组相比,Ad-aMMP-2组MMP-2 mRNA表达下降,MMP-2蛋白分泌减少,明胶酶裂解活性降低(P<0.05).结论 反义MMP-2 cDNA可以有效抑制体外培养增殖期VEC分泌MMP-2.     

碘促进血管内皮细胞增殖作用与MEK1相关性及碘对血管内皮细胞表达VEGF影响的实验研究

目的研究碘促进血管内皮细胞(VEC)增殖的机制.方法(1)使用丝裂原活化蛋白激酶激酶1(MEK1)抑制剂作用与含有碘离子(I-)培养环境中的VEC,以CCK-8法检测细胞增殖情况;(2)在不同浓度环境中培养VEC,使用免疫印迹法和RT-PCR法检测血管内皮生长因子(VEGF)表达情况.结果MEK1抑制剂作用于含有I-培养环境中的VEC后,细胞相对增殖率低于KI组(P<0.01).不同浓度的I-作用于VEC后,VEGF表达未出现上调(P>0.05).结论碘促进血管内皮细胞增殖可能与MEK1激活有关,碘不促进VEC合成VEGF.     

同种异体单核细胞活化血管内皮细胞产生IP-10、I-TAC的体外研究

目的  研究单核细胞对异体血管内皮细胞（VEC）的活化作用以及产生干扰素诱导蛋白10（IP-10）、干扰素诱导的T细胞α型趋化因子（I-TAC）的效果。方法  人外周血中分离单核细胞，酶消化法分离人主动脉内皮细胞，建立单核细胞与VEC共培养系统。用流式细胞仪（FACS）检测VEC表面粘附分子CD54、CD62E表达的情况，用RT-PCR检测VEC内I-TAC和IP 10 mRNA的表达情况。结果  RT-PCR 检测结果表明，VEC与同种异体单核细胞共培养24h后，细胞内IP-10和I-TAC mRNA表达水平显著增加（P<0.05），且在48、72h的表达维持在较高的水平。 FACS检测结果表明，正常培养的VEC少量表达CD54分子，不表达CD62E分子。与同种异体单核细胞共培养24h后，VEC表面CD62E和CD54的表达水平明显上调（P<0.05）。结论  在同种异体单核细胞 血管内皮细胞免疫反应中，单核细胞能活化血管内皮细胞，使内皮细胞表达IP-10、I-TAC等趋化因子和CD54、CD62E等粘附分子，参与对移植器官的排斥反应。     

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.