解偶联蛋白2在PM2.5所致的氧化应激损伤心肌细胞中的作用机制探讨

目的:初步探讨解偶联蛋白2(UCP2)在PM2.5致心肌细胞氧化应激损伤中的作用。方法:培养大鼠心肌细胞株H9C2细胞,分为NC组、PM2.5组、PM2.5+si RNA组三组,分别给予常规的培养基、PM2.5培养基、si RNA+PM2.5培养基刺激,48 h后比色法检测线粒体的活性氧(ROS)、丙二醛(MDA)含量、谷胱甘肽(GSH)含量以及超氧化物歧化酶(SOD)活性。结果:PM2.5组心肌细胞内ROS、MDA含量、GSH含量、SOD活性分别为(69.2±6.3)U/Well、(68.33±1.96)nmol/mgprot、(533.05±10.83)mg/gprot、(50.37±1.98)U/mgprot,PM2.5+si RNA组心肌细胞内ROS(90.2±6.2)U/Well明显增多、MDA含量(88.44±1.27)nmol/mgprot明显升高,GSH含量(421.17±16.90mg/gprot)明显减少,SOD活性(30.09±2.02)U/mgprot明显降低。三组两两比较差异均有统计学意义(P<0.05)。结论:PM2.5可以通过氧化应激途径损伤心肌细胞,RNA干扰UCP2基因表达后,PM2.5致氧化应激增强所介导的心肌细胞损伤加剧,这提示UCP2在PM2.5致氧化应激心肌损伤过程中可能起着保护性作用。     

NU7026抑制DNA-PKcs对辐射旁效应中L-02细胞增殖、凋亡、抗氧化的影响

目的探讨NU7026抑制DNA-PKcs对辐射旁效应中L-02细胞增殖、凋亡、抗氧化的影响。 方法构建辐射旁效应模型。实验分为空白组(阴性对照)、旁效应组(辐射条件培养液处理)、联合组(辐射条件培养液+NU7026)、辐照组(直接辐照)。采用MTT法、克隆形成实验、微核实验、活性氧(ROS)试剂盒、抗氧化活性试剂盒、荧光分光光度法、流式细胞术、Western blotting分别检测细胞存活率、克隆形成率、微核率、活性氧水平、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)活性、丙二醛(MDA)含量、线粒体膜电位、细胞凋亡和周期阻滞及相关蛋白的表达。 结果与空白组比较,其他3组的细胞存活率、克隆形成率、SOD和GSH活力、线粒体膜电位降低(P＜0.05)；微核率、MDA、ROS含量、凋亡率及G2/M期阻滞升高(P＜0.05)。与旁效应组比较,联合组、辐照组的细胞存活率、克隆形成率、SOD和GSH活力、线粒体膜电位降低(P＜0.05)；微核率、MDA、ROS含量、凋亡率及G2/M期阻滞升高(P＜0.05)。 结论NU7026抑制DNA-PKcs能降低辐射旁效应中L-02细胞增殖、抗氧化和抗凋亡的能力。     

糖基化终末产物对SH-SY5Y细胞氧化应激及凋亡的影响

目的研究糖基化终末产物(AGEs)对人神经母细胞瘤细胞(SH-SY5Y)凋亡的影响,探讨AGEs与阿尔茨海默病(AD)发病机制的关系。方法分别以不同浓度糖基化修饰的牛血清白蛋白(AGE-BSA)处理SH-SY5Y细胞48 h,选取AGE-BSA敏感浓度(200μg/mL)处理细胞不同时间,流式细胞仪(FCM)检测细胞凋亡率,确定AGE-BSA诱导SH-SY5Y凋亡最佳浓度及时间。将细胞随机分为对照组、牛血清白蛋白(BSA)对照组、AGE-BSA组、AGE-BSA+抗RAGE中和抗体组、AGE-BSA+α-硫辛酸组、AGE-BSA+DPI(NADPH氧化酶抑制剂)组。FCM检测细胞凋亡率变化,Hoechst 33258染色观察细胞形态改变,应用活性氧荧光探针DCFH-DA检测活性氧(ROS)水平,Western blot测定Caspase-12的表达。结果细胞凋亡率随AGE-BSA浓度增加及作用时间延长而升高,200μg/mL AGE-BSA作用48 h细胞凋亡率从对照组的(2.23±0.08)%增加到(16.8±1.27)%(P<0.05),同时,Hoechst染核后可见典型凋亡形态改变,细胞内ROS水平显著增高,Caspase-12蛋白表达明显上调,与对照组相比差异显著(P<0.05),而分别用抗RAGE中和抗体、α-硫辛酸、DPI预处理后再加入AGE-BSA,各组与AGE-BSA组比较,凋亡率明显下降,ROS水平、Caspase-12蛋白表达均明显降低(P<0.05)。结论糖基化终末产物可刺激SH-SY5Y细胞产生大量ROS及活化Caspase-12介导细胞凋亡,通过阻断其与特异性受体RAGE结合或减少细胞内ROS可减轻细胞凋亡的发生。     

利福平对鱼藤酮诱导的神经细胞SH-SY5Y凋亡的抑制作用

目的 探讨利福平对鱼藤酮所诱导的人多巴胺能神经母细胞瘤细胞SH-SY5Y凋亡的抑制作用.方法 采用100 nmol/L鱼藤酮作用SH-SY5Y细胞24 h,建立SH-SY5Y细胞凋亡模型;再用100、200、300 μmol/L不同浓度的利福平进行干预,采用MTT比色法检测细胞活性,采用流式细胞术(FCM)及原位缺口末端标记染色法(TUNEL)检测细胞凋亡, Western blot法检测前凋亡蛋白3-磷酸甘油醛脱氢酶(GAPDH)的表达水平.结果 用100 nmol/L鱼藤酮作用SH-SY5Y细胞24 h,能够诱导该细胞产生凋亡并降低其活性;利福平干预后,鱼藤酮所诱导的SH-SY5Y细胞凋亡被抑制,并且这一抑制作用与利福平浓度有明显的剂量-效应关系;而且利福平干预后能降低GAPDH蛋白的表达水平.结论 利福平对鱼藤酮诱导的SH-SY5Y细胞凋亡具有明显的抑制作用,这一抑制作用可能是通过下调GAPDH蛋白的表达而得以实现.     

少突胶质样细胞条件培养液通过调节Sirt1/PGC-1α通路改善SH-SY5Y细胞氧化应激损伤

目的:研究人神经干细胞来源的少突胶质样细胞条件培养液(oligodendrocytes-like conditioned medium, OCM)对人神经元氧化应激损伤的修复作用及可能机制。方法:人神经干细胞分化为少突胶质样细胞(oligodendrocytes-like, OLs),免疫荧光等方法鉴定OLs,体外观察其髓鞘形成能力。构建H₂O₂诱导的人神经母细胞瘤细胞(SH-SY5Y)氧化损伤模型,流式细胞术测定活性氧水平及细胞凋亡,荧光探针检测线粒体膜电位,化学发光法检测还原型谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平。蛋白质印迹法检测Sirt1/PGC-1α通路蛋白水平。结果:人神经干细胞源少突胶质样细胞(human neural stem cell-derived oligodendrocytes-like, hNSC-OLs)呈现典型的少突胶质细胞形态,表达少突胶质细胞标志物Olig2和SOX10,在体外包裹轴突形成髓鞘。OCM处理显著提高了受损SH-SY5Y细胞活力(P<0.01),活性氧水平明显降低...     

叶酸对铝诱导的人神经母细胞瘤细胞活性氧和线粒体膜电位水平的影响

目的 探讨叶酸对铝诱导的人神经母细胞瘤细胞(SH-SY5Y细胞)活性氧(ROS)和线粒体膜电位(MMP)水平的影响.方法 叶酸与不同金属共同处理SH-SY5Y细胞24 h,不同浓度Al分别染毒SH-SY5Y细胞6 h,不同浓度叶酸干预Al诱导SH-SY5Y细胞6 h,检测上述各组细胞ROS水平.测定叶酸对Al诱导SH-SY5Y细胞的毒性、MMP水平.结果 与control组比较,300μmol/L Al诱导SH-SY5Y细胞的ROS水平最高(P<0.05).50μmol/L叶酸+300μmol/L Al组细胞DCF相对荧光值、MMP降低的细胞比例、细胞存活率分别为15926±8200、38.04％、82.7％,与300μmol/L Al组20118±12503、45.88％、71.9％比较差异有统计学意义(P<0.05).结论 叶酸可能通过拮抗铝诱导的SH-SY5Y细胞ROS水平升高来阻止MMP的降低,从而提高铝诱导下SH-SY5Y细胞的存活率.     

邻苯二甲酸二乙基己酯对体外培养大鼠睾丸支持细胞氧化应激的 影响

目的对体外培养的大鼠睾丸支持细胞进行邻苯二甲酸二乙基己酯(DEHP)染毒,探讨DEHP对体外培养大鼠睾丸支持细胞氧化应激和细胞凋亡的影响。方法体外分离和原代培养大鼠睾丸支持细胞,不同浓度的DE-HP(10、50、100 nmol·L-1)作用于支持细胞24 h,应用流式细胞术检测支持细胞凋亡率和细胞内活性氧(ROS),应用紫外分光光度计测定细胞培养液中超氧化物歧化酶(SOD)和丙二醛(MDA)含量。结果与对照组相比,DEHP各剂量组大鼠睾丸支持细胞MDA含量和ROS增高(P<0.05),SOD活性降低(P<0.05),细胞凋亡率增加(P<0.05),且呈浓度依赖关系。结论 DEHP可通过激活细胞氧化应激而诱导大鼠睾丸支持细胞凋亡,进而影响支持细胞功能。     

何首乌提取物对MPP+诱导的SH-SY5Y细胞损伤的保护作用

目的 研究何首乌提取物对1-甲基-4-苯基-吡啶离子(MPP+)诱导SH-SY5Y细胞损伤的保护作用.方法 体外培养SH-SY5Y细胞,建立MPP+致SH-SY5Y细胞损伤模型,实验分为对照组、MPP+损伤模型组、何首乌提取物干预组.干预组在MPP+损伤的基础上,给予不同浓度何首乌提取物(终质量浓度5、25、100 mg/L).MTT比色法检测细胞活力;Hoechst33258染色和白光下观察细胞形态变化,比色法检测细胞培养上清液中乳酸脱氢酶(LDH)及细胞中丙二醛(MDA)含量;通过荧光强度观察活性氧(ROS)的含量和线粒体膜电位的变化.结果 与对照组相比,模型组中细胞活性显著降低,Hoechst33258染色和白光下可见细胞胞体变小、核皱缩、碎裂有明显的颗粒状和固缩状荧光,并且细胞上清液中LDH泄漏明显增多、细胞中MDA含量和ROS显著升高而线粒体膜电位显著降低(P＜0.05).与模型组比较,预先4h给予不同浓度的何首乌提取物(5、25、100 mg/L)能明显改善SH-SY5Y细胞的活性,降低细胞产生的LDH、MDA含量,并恢复线粒体膜高能电位,且对ROS有明显的抑制效果(P＜0.01).结论 何首乌提取物对Mpp+诱导的SH-SY5Y细胞损伤具有显著的保护作用,其保护作用与其抗氧化应激作用有关.     

人参首乌方对缺氧复氧诱导的皮质神经细胞凋亡的影响

目的 探讨人参首乌方(RSSW)降低缺血性脑损伤、保护神经元的作用及机制.方法 原代培养大鼠皮质神经元,Na2 S2 O4造成缺氧/复氧损伤模型,分为正常组、模型组、依达拉奉阳性药组、RSSW高、中、低剂量组,通过Hoechst 33258染色及流式细胞术测定细胞凋亡率,生化测定细胞内内源性超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、活性氧(ROS)、过氧化产物丙二醛(MDA)和线粒体膜电位(MMP)及细胞培养上清液乳酸脱氢酶(LDH)水平,ELISA检测细胞内凋亡蛋白Caspase-3,9的活力,Western blot检测其抗凋亡蛋白Bcl-2的表达.结果 RSSW能升高细胞SOD活性,提高GSH及MMP水平,减少细胞内ROS、MDA生成和LDH漏出,抑制Caspase-3,9的活性,具有显著的抗氧化活性,上调Bcl-2蛋白的表达,剂量依赖性抑制神经细胞元凋亡.结论 RSSW对氧化应激损伤诱导的神经细胞凋亡有保护作用,其机制可能与其增加细胞内源性抗氧化剂、改善Bcl-2蛋白的表达及抑制Caspase-3和Caspase-9的活性相关.     

齐墩果酸对oxLDL诱导人脐静脉内皮细胞氧化损伤的保护作用

目的探讨齐墩果酸对氧化性低密度脂蛋白（oxLDL）诱导人脐静脉内皮细胞（HUVEC）损伤的保护作用。方法体外培养HUVEC,以维生素E（200μmol/L）或齐墩果酸（5-40μmol/L）预处理细胞24h,再加入200mg/L oxLDL继续培养24h造成细胞损伤。采用MTT法检测各组细胞存活率,同时检测细胞丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性、还原型谷胱甘肽（GSH）含量,采用DCFH-DA探针法检测细胞内活性氧（ROS）含量。结果与正常组比较,200mg/L oxLDL组细胞存活率、SOD活性、GSH含量明显降低,MDA、ROS含量明显增加,差异均有统计学意义（F=67.72-173.25,q=11.46-30.86,P〈0.01）;与200mg/L oxLDL组比较,齐墩果酸在5-20μmol/L浓度范围内,可剂量依赖性提高细胞存活率、减少MDA含量及ROS形成、增加GSH含量及SOD活性（q=4.26-22.35,P〈0.05）。结论齐墩果酸能抑制ox-LDL诱导的HUVEC氧化损伤,其机制可能与维持SOD活性,减少ROS释放、MDA生成及GSH消耗有关。     

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.