血管内皮细胞脂质过氧化对中膜平 滑肌细胞增殖的影响

本实验以氢过氧化枯烯引发培养血管内皮细胞（ＥＣ）的脂质过氧化反应，观察其条件培养液对中膜平滑肌细胞（ＳＭＣ）ＤＮＡ合成的影响，探讨脂质过氧化作用通过活化内皮细胞促进和加重体内动脉粥样硬化（ＡＳ）病变形成的机理。一、材料及方法１．牛主动脉内皮细胞及中膜平滑肌细胞的培养：于无菌条件下取初生牛主动脉，以消化法用蛋白酶Ｄｉｓｐａｓｅ消化采取内皮细胞，用含１５％胎牛血清（ＦＢＳ）的ＤＭＥＭ培养液培养。第２～５代细胞用于实验。再取牛主动脉中膜，采用贴块法进行原代培养，第３－１０代细胞用于实验。２．内皮细胞脂…     

机械刮取法体外培养牛胸主动脉内皮细胞

目曲：探讨牛胸主动脉内皮细胞（CTAECs）的原代培养方法,提高体外分离培养血管内皮细胞的成功率,为体外研究血管内皮细胞提供实验基础。方法：无菌手术刀片在牛胸主动脉内膜刮取内皮细胞,“力度梯度标记法”探索刮取力度,细胞收集于含20％南美胎牛血清的培养基中培养,对培养细胞进行Ⅷ因子相关抗原免疫细胞化学鉴定。结果：机械刮取法体外培养可以获得大量CTAECs,在倒置显微镜下呈多角形或短梭形,细胞核清晰,呈卵圆形,单层细胞融合后呈铺路石子样排列。细胞生长状态良好,抗Ⅷ因子相关抗原染色呈阳性。结论：机械刮取法是获得CTAECs的一种可取方法,成功率高,可靠性大,可成功构建体外研究血管内皮细胞的模型。     

犬旋股下动脉侧支血管内皮细胞培养的实验研究

目的：探讨犬肢体血管内皮细胞简易培养方法，为在肢端复合组织体外保存方法及保存后组织细胞（血管内皮细胞（活性判定方面的研究奠定基础；方法：手术切取犬旋股下动脉侧支，采用血管外翻术使血管内皮朝外，以０．２０％Ｔｒｙｐｓｉｎ－ＥＤＴＡ液消化分离血管内皮细胞后体外静置培养。利用免疫组化法对增殖细胞分别进行Ⅷ因子和ＣＤ３４相关抗原的检测。结果：分离的细胞活率为９９．３&;#177;０．７％。培养２４小时多数细胞已贴壁，     

牛主动脉内皮细胞的分离、培养及特征

目的:建立牛主动脉内皮细胞(BAEC)的体外研究模型,为体外研究内皮细胞的病理和生理过程提供重要手段.方法:用酶消化法分离BAEC,在pH为7.2的DMEM培养基中培养,鉴定内皮细胞中特异表达的第Ⅷ因子;以计数法测细胞的增殖能力,同时测定细胞的迁移.结果:细胞呈三角形,单层生长,第Ⅷ因子表达阳性,增殖呈指数生长,迁移正常.结论:提示牛主动脉内皮细胞的体外模型建立成功.     

大鼠肺微血管内皮细胞的G蛋白亚型鉴定

本实验拟在体外培养大鼠肺微血管内皮细胞 (ＲＰＭＶＥＣｓ)的基础上 ,用免疫细胞化学Ｓ Ｐ法和免疫荧光细胞化学间接法检测ＲＰＭＶＥＣｓ膜Ｇ蛋白亚型。1　材料和方法1.1　材料　　ＤＭＥＭ培养干粉 (美国ＧＩＢＩＣＯ公司 ) ;荧光 (ＦＩＴＣ)结合的羊抗兔抗体、兔抗大鼠ＩｇＧ抗体免疫血清 (北京中山公司 ) ;Ｇｓα、Ｇｉα和Ｇｑα兔抗大鼠抗体 (ＩｇＧ) (ＳａｎｔａＣｒｕｚＢｉｏｔｅｃｈｎｏｌｏｇｙ .Ｉｎｃ)。1.2　检测ＲＰＭＶＥＣｓＧ蛋白亚型方法1.2 .1　免疫细胞化学Ｓ Ｐ法　　ＲＰＭＶＥＣｓ经 4%多聚甲醛固定 ,3 %Ｈ2 Ｏ2 …     

大鼠肺动脉内皮细胞的简易培养鉴定

目的：探索一种简单的方法培养大鼠肺动脉内皮细胞（ＲＰＡＥＣｓ）方法：分离大鼠肺动脉，将共切成步块，使其内皮面贴于培养瓶，用含有２０％新生牛血清，肝索９０μｇ／ｍｌ，Ｌ－谷氨酰胺４ｍｍｏｌ／Ｌ，青霉素１００ｕ／ｍｌ和链霉素１００ｕｇ／ｍｌ的ＲＰＭＩ－１６４０培养基培养，结果：培养２４ｈ后ＲＰＡＥＣｓ从动脉块游出，７２ｈ后将动脉块移去，ＲＰＡＥＣｓ于６～１０ｄ融合成单层细胞，这些细胞具有规律的鹅卵石样     

切应力对内皮细胞一氧化氮合酶及细胞凋亡的影响

目的探讨切应力作用对体外培养的牛胸主动脉内皮细胞一氧化氮合酶(eNOS)蛋白表达和细胞凋亡的影响。方法体外培养牛胸主动脉内皮细胞，采用平行平板流动腔装置模拟体内血流切应力(5dynes／cm^2和25dynes/cm^2)作用于内皮细胞12h，采用Western blot分析和DNA片段凝胶电泳，观察不同切应力对体外培养的血管内皮细胞eNOS蛋白表达和细胞凋亡的影响。结果静息状态下，体外培养的牛胸主动脉内皮细胞呈多角形“铺路石”状排列，形态大小均匀。不同切应力(5，25dyne／cm^2)作用12h后，血管内皮细胞形态学未见明显改变。但与静止状态细胞的基础水平相比，Western blot分析发现，牛胸主动脉血管内皮细胞eNOS蛋白表达在低切应力时降低；低切应力还可诱导血管内皮细胞发生凋亡，凝胶电泳检测出现明显的DNA梯形带；但在高切应力(25dynes／cm^2)作用下，内皮细胞eNOS表达水平与未处理组接近，而且DNA片段凝胶电泳也未见明显凋亡现象。结论低切应力可降低内皮细胞eNOS蛋白表达和诱导内皮细胞凋亡，提示低切应力可能是导致血管内皮细胞eNOS表达降低和细胞凋亡的重要因素。     

小鼠内皮抑素基因真核表达载体的构建和表达

目的 构建小鼠内皮抑素（ｅｎｄｏｓｔａｔｉｎ）ｃＤＮＡ的真核表达载体并将其在体外培养的脑胶质瘤细胞内表达。方法 将带有分泌信号的小鼠ｅｎｄｏｓｔａｔｉｎｃＤＮＡ克隆入真核表达载体ｐｃＤＮＡ３,构建ＣＭＶ启动子控制的载体ｐｃＤＮＡ－ｓＥｎｄｏ,并将上述载体转染体外培养的脑胶质瘤细胞（ＳＨＧ４４）,采用ＷｅｓｔｅｒｎＢｌｏｔ鉴定其表达,应艇牛微血管内皮细胞抑制实验检测其活性,结果 成功地构建了真核表达     

大鼠乳鼠腹主动脉血管内皮细胞的培养与鉴定

目的：研究大鼠乳鼠腹主动脉血管内皮细胞的培养与鉴定方法。方法：取出生5~7d左右的大鼠乳鼠的腹主动脉血管,用组织块法在含20%胎牛血清及内皮细胞生长添加剂和肝素的DMEM培养基中培养,倒置相差显微镜观察内皮细胞形态和生长特征,免疫荧光方法进行血管内皮细胞Tie2相关抗原的鉴定。结果：来自于大鼠乳鼠腹主动脉的血管内皮细胞在改良的培养基中进行体外培养,能够保持增殖和传代的能力,且Tie2抗原鉴定呈阳性。结论：成功分离大鼠乳鼠腹主动脉组织并且获得体外原代培养大鼠乳鼠腹主动脉内皮细胞。用组织培养方法可以获得较满意的血管内皮细胞。     

随机贴块法培养大鼠主动脉内皮细胞及其鉴定

目的 寻找一种培养及鉴定大鼠主动脉内皮细胞的简便方法.方法 采用随机贴块法培养大鼠主动脉内皮细胞并用差速消化法加以纯化,观察细胞形态学特性,用抗vwF抗体免疫荧光染色及内皮细胞体外成管试验鉴定.结果 培养3d即有细胞生长,1周可融合成层.抗vwF抗体免疫荧光染色及内皮细胞体外成管试验阳性.结论 本方法简便易行,适于推广应用.     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.