混合脐血浆在人脐静脉内皮细胞培养中的应用

目的：观察混合脐血浆在人脐静脉内皮细胞（ｈｕｍａｎｕｍｂｉｌｉｃａｌｖｅｉｎｅｎｄｏｔｈｅｌｉａｌｃｅｌｓ,ＨＵＶＥＣ）培养中的作用。方法：在含２０％（体积分数）混合脐血浆的内皮细胞培养体系中进行ＨＵＶＥＣ的原代和传代培养,并对培养细胞进行纯度鉴定。同时用噻唑蓝试验观察不同体积分数的混合脐血浆对ＨＵＶＥＣ增殖活力的影响。结果：２０％（体积分数）的混合脐血浆对ＨＵＶＥＣ培养有明显支持作用,该体系培养的ＨＵＶＥＣ经第Ⅷ因子相关抗原（Ⅷ：Ａｇ）和Ⅰ型荆豆凝集素（ＵＥＡⅠ）鉴定纯度可达９３．１％。结论：２０份混合的脐血浆可以代替内皮细胞生长因子成功地用于ＨＵＶＥＣ的培养,以获得高纯度可传代的内皮细胞。     

混合脐血浆在人脐静脉内皮细胞培养中的应用

目的：观察混合脐血浆在人脐静脉内皮细胞（ｈｕｍａｎ ｕｍｂｉｌｉｃａｌ ｖｅｉｎ ｅｎｄｏｔｈｅｌｉａｌ ｃｅｌｌｓ，ＨＵＶＥＣ）培养中的作用。方法：在含２０％（体积分数）混合脐血浆的内皮细胞培养体系中进行ＨＵＶＥＣ的原代和传代培养，并对培养细胞进行纯度鉴定。同时用噻唑蓝试验观察不同体积分数的混合脐血浆对ＨＵＶＥＣ增殖活力的影响。结果：２０％（体积分数）的混合脐血浆对ＨＵＶＥＣ２有明显支持作用，该     

氧化修饰LDL对培养人脐胸脉内皮细胞DNA合成和胆固醇酯合成…

目的：研究氧化低密度脂蛋白（ｏｘＬＤＬ）对内皮细胞的损伤作用。方法：以胸腺嘧啶核苷（＾３Ｈ－ＴｄＲ）掺入法和胆固醇酯薄层层析法，观察ｏｘＬＤＬ对培养的人脐静脉内皮细胞（ＨＵＮＥＣ）ＤＮＡ合成和胆固醇酯含量的影响。结果：ｏｘＬＤＬ抑制内皮细胞ＤＮＡ合成，并与浓度及作用时间相关，１００ｍｇ．Ｌ＾－１的ｏｘＬＤＬ作用２４ｈ的内皮细胞掺入＾３Ｈ－ＴｄＲ的ｄｐｍ数为不加ＬＤＬ组的４８％（Ｐ〈０．００１），天     

小鼠脑微血管内皮细胞的体外培养

目的 探讨脑微血管内皮细胞的体外培养方法并进行细胞超微结构研究及组织型纤溶酶原激活物（ＴＰＡ）活性测定。方法 取新生小鼠脑组织,通过匀浆、过筛、胶原酶消化、差速粘附等技术对鼠脑微血管内皮细胞进行原代培养,待细胞铺满瓶底时,用０．１２５％胰酶－０．０２％ＥＤＴＡ消化,离心收集内皮细胞,进行传代培养。原代、传代各取８例,吸取培养液用酶联免疫吸附试验测试ＴＰＡ活性。结果 经Ⅷ因子相关抗原免疫组织化学鉴定     

益气活血方对自由基介导肿瘤坏死因子损伤血管内皮细胞的影响

以体外原代培养的人脐静脉内皮细胞（ＨＵＥＣ）为试验对照，观察肿瘤坏死因子（ＴＮＦ）作用于ＨＵＥＣ后，细胞形态及细胞内超氧化物歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）和脂质过氧化物（ＬＰＯ）的变化及中药益气活血方的作用。结果显示：与空白对照组比较，ＴＮＦ组细胞内ＳＯＤ活性下降而ＬＰＯ含量升高（Ｐ＜０．０１），细胞收缩，核质密度增大，益气活血方（２０ｍｇ／ｍｌ）能抑制ＴＮＦ引起的ＳＯＤ活性下降和ＬＰＯ含量升高（Ｐ＞０．０５），并保护其形态的过度改变，提示抗脂质过氧化可能是益气活血方抗ＴＮＦ损伤血管内皮细胞的重要机制。     

人脐静脉内皮细胞的体外培养及鉴定

目的：探讨人血管内皮细胞体外培养方法,为研究心脑血管疾病发病机制提供实验手段。方法：采用0．1％胶原酶消化、分离体外原代培养脐静脉血管内皮细胞,0．125％胰酶-0．01％EDTA 溶液消化传代。并用光镜和免疫组化等方法进行内皮细胞的形态观察和鉴定。结果：原代培养的内皮细胞在接种后约4h开始贴壁生长,光镜下内皮细胞呈铺路石状镶嵌排列;免疫组化可见内皮细胞胞浆中人第Ⅷ因子相关抗原呈阳性反应。结论：用胶原酶灌注消化脐静脉可获得高纯度的内皮细胞,细胞存活率高,为血管内皮细胞的研究提供了实验模型。     

氧化修饰LDL对培养人脐静脉内皮细胞DNA合成和胆固醇酯合成的影响

研究氧化低密度脂蛋白（ｏｘＬＤＬ）对内皮细胞的损伤作用。方法：以胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入法和胆固醇酯薄层层析法，观察ｏｘＬＤＬ对培养的人脐静脉内皮细胞（ＨＵＶＥＣ）ＤＮＡ合成和胆固醇酯含量的影响。结果：ｏｘＬＤＬ抑制内皮细胞ＤＮＡ合成，并与浓度及作用时间相关，１００ｍｇ·Ｌ－１的ｏｘＬＤＬ作用２４ｈ的内皮细胞掺入３Ｈ－ＴｄＲ的ｄｐｍ数为不加ＬＤＬ组的４８％（Ｐ＜０．００１），天然ＬＤＬ（ｎＬＤＬ）对ＨＵＶＥＣ掺入３Ｈ－ＴｄＲ也有轻度的抑制作用。ｏｘＬＤＬ降低ＨＵＶＥＣ内胆固醇酯的含量，５０ｍｇ·Ｌ－１ｏｘＬＤＬ孵育的ＨＵＶＥＣ胞内胆固醇酯含量是ｎＬＤＬ组的４２％。扫描电镜发现ｏｘＬＤＬ孵育的ＨＵＶＥＣ形态发生改变，胞间连接减少。结论：ｏｘＬＤＬ对ＨＵＶＥＣ具有细胞毒作用，促发动脉粥样硬化。     

人脐静脉内皮细胞的培养及鉴定

目的：摸索人脐静脉内皮细胞体外培养的方法和注意事项。方法：采用0．25％胰蛋白酶分离人脐静脉内皮细胞,含20％FBSDMEM养基培养,待细胞80％融合后,以O．25％胰蛋白酶消化传代,并以倒置显微镜观察内皮细胞的生长情况,以免疫组化方法对内皮细胞进行鉴定。结果：原代培养的细胞在接种4h后开始贴壁生长,3－5天后融合成单层,倒置显微镜下观察细胞呈铺路石状排列,免疫组化显示人第Ⅷ因子相关抗原阳性。结论：用胰蛋白酶消化脐静脉可获得内皮细胞且成活率较高,是一种可靠的方法。     

糖基化低密度脂蛋白对血管内皮的损伤作用

目的探讨糖基化低密度脂蛋白对血管内皮的实验性损伤作用。方法以体外非酶糖基化低密度脂蛋白与人脐静脉内皮细胞（ＥＣ）共同培养，以低密度脂蛋白（ＬＤＬ）作对照，观察内皮细胞结构及功能的改变。结果糖化ＬＤＬ２５０μｇ／ｍｌ可引起ＥＣ超微结构改变，增殖率减低（Ｐ＜０．０５）。培养上清液中血管紧张素转化酶（ＡＣＥ）活性降低（Ｐ＜０．０５），ＶｏｎＷｉｌｌｂｒａｎｄＦａｃｔｏｒ（ＶＷＦ）值升高（Ｐ＜０．００１），而糖化ＬＤＬ１２５μｇ／ｍｌ、７５μｇ／ｍｌ与ＬＤＬ对ＥＣ的作用无差异。结论在体外，糖基化ＬＤＬ２５０μｇ／ｍｌ可引起ＥＣ损伤，糖化ＬＤＬ在体外对触发动脉粥样硬化的发生、发展有重大意义。     

白细胞介素6,肿瘤坏死因子对体外培养的血管内皮细胞的损伤 …

目的：研究白细胞介素６（ＩＬ－６）和肿瘤坏死因子（ＴＮＦ）对人脐静脉内皮细胞（ＨＶＥＣ）的损伤作用。方法：以原代培养的ＨＶＥＣ为模型,将ＩＬ－６、ＴＮＦ及其抗体与ＨＶＥＣ共同孵育,观察内皮细胞（ＥＣ）形态变化和培养液中血管紧张素转化酶（ＡＣＥ）和乳酸脱氢酶（ＬＤＨ）水平。结果：ＴＮＦ和ＩＬ－６可引起间隙增大,甚至细胞脱落;并且随着ＴＮＦ和ＩＬ－６浓度加大及作用时间延长,培养液中ＡＣＥ和ＬＤＨ水平长     

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.     

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.