hexarelin对ox-LDL引起的大鼠脑微血管内皮损伤的保护作用

目的建立体外培养大鼠脑微血管内皮细胞ox-LDL损伤模型,探讨hexarelin对大鼠脑微血管内皮细胞的保护作用。方法分离并培养大鼠脑微血管内皮细胞,分成正常对照组（control）、ox-LDL组（ox-LDL）、hexarelin ＋ ox-LDL组（hex＋ox-LDL）3组。其中ox-LDL组在培养液中加入ox-LDL 50mg/L; hex＋ox-LDL组加入ox-LDL 50mg/L和0.1μmol/L hexarelin;正常对照组加入等量PBS。各组细胞无血清培养12h,MTT法检测细胞存活率,TUNEL法检测细胞凋亡情况,Western blot法检测内皮细胞eNOS蛋白表达情况以及蛋白质硝基化情况。结果50mg/L ox-LDL作用12h可明显诱导内皮细胞凋亡,减少eNOS蛋白表达,促进蛋白质的硝基化。而用0.1μmol/L hexarelin和50mg/L ox-LDL共同孵育,可明显减少内皮细胞的凋亡,提高eNOS的表达水平,减少蛋白质的硝基化程度。结论hexarelin能有效抑制由ox-LDL引起的脑微血管内皮细胞损伤,提示hexarelin对脑动脉粥样硬化可能有一定抑制作用。     

卡维地洛对ox-LDL诱导的人主动脉内皮细胞氧化损伤的保护作用

目的：研究卡维地洛（ CAR）对氧化低密度脂蛋白（ ox-LDL）诱导的人主动脉内皮细胞（ HAECs）损伤的保护作用。方法：体外培养HAECs,分为对照组、模型组、CAR高剂量组（ CAR H组）和CAR低剂量（ CAR L组）,即对照组（无血清DMEM）、模型组（200 mg/L ox- LDL 100μL）、CAR高剂量组（ CAR H组,200 mg/L ox-LDL及1×10-5 mol/L CAR各100μL）及CAR低剂量组（ CAR L组,200 mg/L ox-LDL及1×10-6 mol/L CAR各100μL）,后两组 CAR预保护1 h后,加入ox-LDL作用24 h;采用MTT法分析细胞存活率,吉姆萨（ Giemsa）染色后观察HAECs形态;酶联免疫吸附法（ ELISA）检测培养液中一氧化氮（ NO）含量及乳酸脱氢酶（ LDH）外漏量。结果：模型组与对照组相比,细胞发生严重的病理形态学改变,细胞存活率及NO含量显著降低,LDH外漏量显著升高（ P﹤0.05）,CAR H组和CAR L组细胞接近对照组,细胞存活率及NO含量较模型组显著提高,LDH外漏量显著降低（ P﹤0.05）。结论：CAR能减少ox-LDL诱导的HAECs损伤,其作用可能与NO系统有关。     

依帕司他对高糖诱导的血管内皮损伤的保护作用

目的初步研究依帕司他保护高糖诱导的血管内皮细胞损伤的作用。方法体外培养脐静脉血管内皮细胞HUVEC,高糖刺激8 h后利用化学发光法检测细胞培养上清中NO的含量,利用Real time PCR和Western blot分别检测细胞中内皮型一氧化氮合酶(eNOS)的mRNA和蛋白表达水平,以同浓度的甘露醇作为对照。给予依帕司他(0.1μmol/L)预处理30 min,随后再进行高糖培养,检测NO的含量以及eNOS的mRNA和蛋白表达,同时检测依帕司他的靶向基因醛糖还原酶(AR)以及NO的上游基因NADPH氧化酶4(NOX4)的蛋白表达水平。结果高糖培养8 h之后,血管内皮细胞分泌NO下降,eNOS的mRNA和蛋白表达水平下降,与甘露醇对照组相比,具有统计学差异(P<0.05);预先给予依帕司他处理后再进行高糖培养,细胞中eNOS的mRNA和蛋白表达水平均升高,NO表达上调,与单独高糖培养组相比,具有统计学差异(P<0.05)。同时,细胞中AR和NOX4的蛋白表达下降。结论高糖可以诱导内皮细胞损伤,表现为NO的分泌水平降低。依帕司他可能是通过抑制AR和NOX4的表达而发挥对血管内皮细胞损伤的保护作用。     

大豆异黄酮对氧化损伤血管内皮细胞抗氧化作用的研究

目的：探讨大豆异黄酮（SI）对氧化损伤血管内皮细胞的抗氧作用。方法：体外培养血管内皮细胞，将细胞分为5组，即空白对照组（control)，氧化损伤对照组（ox-LDL)，氧化损伤加入维生素E对照组(VC ox-LDL)，氧化损伤加入大豆异黄酮低，中，高浓度组（SI－L＋ox-LDL,SI-M ox-LDL,SI-H ox-LDL)。将氧化低密度脂蛋白（ox-LDL）作用于预先加入VE及不同浓度SI孵育24h的内皮细胞，继续培养24h，测定细胞外及细胞内丙二醛（MDA）含量，超氧化物歧化酶（SOD），谷胱甘肽过氧化物酶（GSH－PX）活性，乳酸脱氢酶（LDH）释放情况及一氧化氮（NO)生成量。结果：ox-LDL组MDA含量，LDH释放百分比高于control组及VE＋ox-LDL组，（SI－L，M，H）＋ox-LDL组，差异有显著性（P＜0．01）；而ox-LDL组SOD，GSH－PX活性及NO浓度均低于control组和VE ox-LDL组，（SI－L，M，H）＋ox-LDL组，差异有显著性（P＜0．01）；另外，SI－M＋ox-LDL组与VE＋ox-LDL组比较，各指标均无显著性差异（P＞0．05）。结论：SI可减轻ox-LDL对血管内皮细胞的氧化损伤，起到一定的抗氧化保护作用。     

胰高血糖素样肽-1对高糖诱导人脐静脉内皮细胞凋亡的影响及相关机制

目的：探讨胰高血糖素样肽-1（glucagon—like peptide-1,GLP—1）对高糖诱导人脐静脉内皮细胞huma numbilica lvein endothelial cells,HUVECs）凋亡的影响及相关机制。方法：HUVECs加入不同处理因素后分组,分别培养48h后,MTT检测细胞活力;流式细胞仪检测细胞早期凋亡率;Westernl印迹测定细胞P-Akt,p-eNOS水平;NO试剂盒检测NO浓度。结果：高糖（33mmol／L）培养48h后HUVECs细胞活力下降p〈0．05）,细胞凋亡率增加（p〈0．01）,细胞p-Akt,p-eNOS,NO水平均下降（P〈0．05）;高糖条件下加人GLP．1（3nmol／L）培养48h,与高糖组相比较,HUVECs细胞活力增加（P〈0．01）,细胞凋亡率减少（p〈O．05）,细胞p-Akt,p-eNOS,NO水平均增／JH（V〈0．05）;P13K抑制剂wortmannine（100nmol／L）可以阻断GLP-1的抗凋亡作用及其对P—Akt蛋白、p-eNOS蛋白及NO水平的影响;eNOS抑制剂L—NAME（100umol／L）仅能阻断GLP-1的抗凋亡作用及对NO水平的影响,不影响p-Akt蛋白的表达。结论：GLP．1可改善高糖诱导的HUVECs凋亡,该抗凋亡作用可能与P13K／Akt／eNOS通路的上调相关。     

非诺贝特对LPC诱导脐静脉内皮细胞增殖、凋亡及eNOS基因表达的影响

目的：探讨非诺贝特对LPC诱导的脐静脉内皮细胞增殖、凋亡及内皮型一氧化氮合酶（eNOS）基因表达的影响。方法：体外培养人脐静脉内皮细胞（HUVECs），根据非诺贝特不同浓度分为正常对照组、LPC组、低浓度非诺贝特（10μmol／L）组、中浓度非诺贝特（50μmol／L）组、高浓度非诺贝特（100μmol／L）组，根据非诺贝特不同干预时间分为正常对照组、LPC组、非诺贝特（50μmol／L）干预6h组、12h组、24h组、48h组进行实验。分别观测不同浓度及不同时间非诺贝特内皮细胞增殖、凋亡、NO浓度及eNOS mRNA表达的变化。结果：与正常对照组比较，LPC抑制内皮细胞增殖，促进细胞凋亡，并使HUVECs eNOS mRNA表达降低，NO合成减少。非诺贝特可干预LPC对内皮细胞的作用，使内皮细胞增殖增强，细胞凋亡减少，eNOS mRNA表达升高，NO合成增加，且其作用呈时间-效应、浓度-效应依赖关系。结论：非诺贝特可改善LPC对HUVECs的影响，使内皮细胞增殖增强，凋亡减少，eNOS mRNA表达升高，NO合成增加，从而起到抗动脉硬化作用。     

五味子甲素SchA缓解MPP＋诱导的SH-SY5Y细胞损伤的机制研究

目的：探讨五味子甲素（SchA ）缓解1-甲基-4-苯基吡啶离子（M PP＋）诱导S H-SY5Y细胞损伤的可能机制。方法实验分为5组,空白对照组,加入无血清培养基进行培养;模型组（M PP＋进行诱导）;SchA组;SchA与M PP＋共同处理的SchA组,其中SchA终浓度分别为1、3、5μmol/L ,M PP＋终浓度为1 mmol/L。NO检测试剂盒检测各组细胞 NO含量,Western blot检测细胞总Akt、p-Akt的蛋白表达情况。结果与对照组相比,1 mmol/L MMP＋诱导后,SH-SY5Y细胞NO含量明显增高（P＜0．05）,经5μmol/L SchA及MPP＋共同处理的SH-SY5Y细胞,NO的含量明显降低。各组总Akt相对表达水平没有明显变化（P＞0．05）;模型组细胞p-Akt相对表达水平较对照组减少,SchA组细胞p-Akt蛋白相对表达水平随SchA浓度的升高而升高,且与模型组比较差异都有统计学意义（P＜0．05）。结论 SchA缓解MPP＋诱导的SH-SY5Y细胞损伤可能与影响NO含量及p-Akt蛋白表达有关。     

氧化低密度脂蛋白(ox-LDL)对人脐静脉内皮细胞(H-UVEC)孤儿核受体NOR1表达的影响

目的:探讨氧化低密度脂蛋白(ox-LDL)对人脐静脉内皮细胞(HUVEC)孤儿核受体NOR1表达的影响。方法:采用ox-LDL建立人脐静脉内皮细胞损伤模型,将细胞分为空白组、ox-LDL高、中、低剂量组(25μg/ml、50μml、100μg/ml、200μg/ml),采用CCK8法检测HUVEC活性,采用实时荧光定量PCR分析NOR1转录水平变化,采用Western-Blot分析NOR1蛋白表达的变化。结果:在HUVEC中加入不同浓度ox-LDL共同培养12h后,CCK8值与ox-LDL呈现明显的剂量反应关系,与空白对照组相比具有显著性差异(P<0.01)。Ox-LDL能够诱导NOR1 mRNA转录和蛋白表达,NOR1 mRNA峰值出现在ox-LDL共同培养后的3h,NOR1蛋白质最大值出现在ox-LDL共同培养后的6 h。结论:ox-LDL能够损伤HUVEC,并能诱导NOR1的表达上调,其在内皮细胞损伤和动脉粥样硬化炎性反应中的作用有待进一步研究。     

氨氯地平对ox-LDL致大鼠骨髓EPCs损伤的保护作用与上调一氧化氮合酶表达有关

目的探讨氨氯地平对氧化型低密度脂蛋白（ox-LDL）致大鼠骨髓内皮祖细胞（EPCs）损伤的保护作用。方法实验分为对照组、ox-LDL组、氨氯地平组（0.1、0.5、1.0μmol/L）,贴壁法培养分离内皮祖细胞,细胞免疫荧光鉴定EPCs,MTT法检测细胞活性,Western blotting检测一氧化氮合酶（eNOS）蛋白表达水平。结果贴壁法能分离纯度达70%以上的CD133＋/VEGFR-2＋双阳性EPCs;0.5μmol/L氨氯地平可显著拮抗ox-LDL对EPCs增殖的抑制作用;0.5μmol/L氨氯地平能显著上调eNOS蛋白表达（P〈0.01）。结论氨氯地平能拮抗ox-LDL对EPCs的损伤,氨氯地平的拮抗作用与上调eNOS表达有关。     

恩格列净通过调控AMPK/eNOS信号通路改善ox-LDL诱导的内皮祖细胞功能障碍

目的 研究恩格列净(EMP)对氧化低密度脂蛋白(ox-LDL)诱导内皮祖细胞(EPCs)损伤的保护作用及机制。方法 通过密度梯度离心法提取、分离并培养小鼠骨髓来源的的EPCs。采用Dil标记乙酰化低密度脂蛋白(Dil-ac-LDL)联合FITC标记荆豆凝集素-1（FITC-UEA-1）双摄取法鉴定。将EPCs分为正常对照组,ox-LDL组以及ox-LDL联合不同浓度恩格列净实验组。CCK-8检测细胞活力,Transwell检测细胞迁移,FITC-Annexin V/PI检测细胞凋亡,ELISA检测细胞上清液中血管内皮细胞生长因子(VEGF)、基质细胞衍生因子-1α(SDF-1α)含量;流式细胞术检测一氧化氮(NO)的合成情况。Western blot检测AMPK、p-AMPK、eNOS、p-eNOS的蛋白表达。结果 提取的EPCs诱导培养至第7天经鉴定为小鼠骨髓EPCs。与对照组比较,ox-LDL组细胞活力降低,迁移细胞减少,凋亡增加,VEGF、SDF-1α含量降低,NO合成减少（P＜0.05）;与ox-LDL组相比,不同浓度恩格列净组细胞活力有所提高,迁移细胞增多,凋亡减少,VEGF、SDF-1α含量升高,NO合成增加（P＜0.05）。ox-LDL处理可明显抑制AMPK及eNOS磷酸化（P＜0.05）,恩格列净处理可以改善AMPK及eNOS磷酸化水平（P＜0.05）,而AMPK抑制剂Compound C可使恩格列净改善EPCs功能活性的作用受到明显的抑制（P＜0.05）。结论 恩格列净可改善ox-LDL诱导的EPCs功能障碍,其机制与调控AMPK/eNOS信号通路有关     

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.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.