AIF对人宫颈癌Hela细胞体外生物学活性的影响及机制研究

目的 探讨凋亡诱导因子(AIF)对人宫颈癌Hela细胞体外生物学活性的影响及其作用机制.方法 通过脂质体转染pcDNA3空载体质粒及pcDNA3-AIF质粒于人宫颈癌Hela细胞,采用免疫荧光反应,Real time PCR及Western blot法检测细胞中AIF的表达.四甲基偶氮唑蓝(MTT)法检测细胞活力,Annexin V-PI流式双染及Hoechst染色检测细胞凋亡情况,Real-time PCR及Western blot法检测细胞中腺苷二磷酸核糖聚合酶-1(PARP-1)及核酸内切酶G(EndoG)蛋白及mRNA的表达.结果 与pcDNA3组比较,peDNA3-AIF组细胞中AIF主要定位于细胞核中,且细胞核中AIF蛋白及mRNA表达量皆显著提高(P＜0.01),同时细胞活力下降,细胞凋亡率提高(P＜0.01),细胞中PARP-1、EndoG蛋白及mRNA表达量显著提高(P＜0.01).结论 AIF具有促进宫颈癌Hela细胞凋亡的作用,可能与PARP-1/AIF/EndoG信号通路有关.     

Puma基因对HeLa细胞凋亡的作用及机制的初步研究

目的 研究外源性puma基因表达对HeLa细胞的作用以及线粒体释放的细胞色素C(Cyt c)和凋亡诱导因子(AIF)是否参与了凋亡过程.方法 脂质体转染HeLa细胞;RT-PCT鉴定表达效果;AO/EB染色检测puma基因的促凋亡活性;Western blot检测细胞凋亡发生后线粒体中Cyt c和AIF位置是否发生迁移.结果 脂质体介导下成功实现了外源性puma基因在HeLa细胞中高表达,AO/EB染色显示转染puma基因后HeLa细胞出现了一系列凋亡形态学改变,这种变化转染48h比24h时更明显;Western blot结果显示,puma基因表达后HeLa细胞的线粒体膜间隙蛋白Cyt c、AIF向胞浆迁移,迁移的蛋白量在转染48h比24h更明显.结论 Puma基因表达有促进HeLa细胞凋亡的作用,且Cyt c和AIF参与puma基因促进HeLa细胞凋亡的过程.     

四逆汤抗阿霉素性心衰心肌细胞线粒体凋亡机制探讨

目的探讨四逆汤抗阿霉素（ADR）性心衰大鼠心肌细胞凋亡的作用及线粒体机制。方法SD大鼠随机分为对照组,心衰组和四逆汤组,心衰组和四逆汤组尾静脉注射ADR复制心衰模型,四逆汤组给予四逆汤煎剂灌胃（3．75g生药／kg/d）,实验结束时留取各组大鼠心肌组织,检测线粒体丙二醛（MDA）含量,MnSOD活性及其肿胀程度;Western blotting检测胞浆中细胞色素C（Cyt-C）、凋亡诱导因子（AIF）表达;比色法测定Caspase-3、Caspase-9蛋白酶的活性;流式细胞术测定心肌细胞凋亡情况。结果四逆汤组可以提高线粒体MnSOD的活性,减少MDA的含量及肿胀程度;有效抑制Cyt—C及AIF蛋白释放及Caspase-9和Caspase-3蛋白酶活性,减少凋亡的发生。结论线粒．体在阿霉素性心衰发病中可能占有重要作用,四逆汤可能是通过减轻线粒体氧化损伤,减少心肌细胞凋亡来保护心肌组织。     

PUMA基因对HELA的作用及其机制的初步研究

目的 初步探讨PUMA基因对HELA细胞是否具有促凋亡的活性和其参与HELA细胞凋亡的过程线粒体中cyt-c、AIF位置足否发生迁移.方法 倒置相差显微镜、AO/EB染色检测PUMA基因的促凋亡活性;Western blot检测HELA细胞凋亡后线粒体中cyt-c、AIF位置是否发生迁移.结果 ①倒置相差显微镜和荧光显微镜结果显示转染PUMA基因后HELA细胞出现了一系列凋亡形态学改变,这种变化在转染48h比24h更明显;②Western blot结果显示HELA细胞凋亡后线粒体内膜腔隙蛋白cyt-c、AIF向胞质迁移,迁移的蛋白量在转染48h比24h更明显.结论 ①PUMA基因有促进HELA细胞凋亡的作用;②AIF和cyt-c参与PUMA基因促进HELA细胞凋亡的过程,但此过程有无涉及线粒体膜电位的变化还有待进一步的研究.     

PARP-1激活和AIF易位在肠缺血再灌注损伤中的作用

目的:研究大鼠肠缺血再灌注后肠损伤中是否存在聚腺苷二磷酸核糖聚合酶-1(PARP-1)激活和凋亡诱导因子(AIF)易位,并探索二者在肠缺血再灌注损伤中的作用?方法:通过阻断腹腔干和肠系膜上动脉30 min后,恢复血运,制作鼠肠缺血再灌注模型,分为肠缺血再灌注组(I/R),缺血再灌注前15min经静脉给PARP-1抑制剂3-AB组(Drug)和假手术组(Sham)?取肠组织分别作HE染色?聚腺苷二磷酸核糖(PAR)的免疫组化染色,TUNEL方法检测组织细胞凋亡情况,Western Blot检测凋亡早期信号PARP-1片段p85和凋亡诱导因子(AIF)的表达情况?结果:I/R组肠损伤程度?PAR阳性表达率?细胞凋亡率和凋亡早期信号PARP-1 p85片段?AIF表达程度均较Sham组显著增高(P < 0.05);应用PARP-1抑制剂后,Drug组上述指标均较I/R组显著降低(P < 0.05),但仍高于Sham组?结论:在大鼠肠缺血再灌注损伤中存在PARP-1激活和AIF易位两分子事件,并在肠缺血再灌注损伤中发挥重要作用;肠缺血再灌注前应用PARP-1抑制剂有肠保护效果?     

10058-F4诱导宫颈癌CaSki细胞凋亡及其机制探索

目的 探讨10058-F4诱导宫颈癌CaSki细胞凋亡的现象及机制.方法 用CCK8的方法检测不同浓度的10058-F4和10058-F4作用不同时间对CaSki细胞生长的影响;流式细胞术检测10058-F4对CaSki凋亡率的影响;Western blot方法检测10058-F4的药物有效性(c-MYC的表达下调)及对CaSki细胞的凋亡诱导作用;Western blot和RT-PCR的方法检测线粒体蛋白的表达情况,流式细胞术检测细胞ROS产生.结果 通过CCK8实验检测出10058-F4对CaSki的增殖抑制具有时间依赖性和浓度依赖性,始于第48小时,IC50为122μmmol/L.用122 μmmol/L的10058-F4处理CaSki细胞48 h,通过流式细胞术检测凋亡发现处理组细胞出现明显凋亡(52％),同时Western blot检测发现剪切型PARP表达量上调.Western blot和RT-PCI检测发现122 μmmol/L的10058-F4处理CaSki细胞48 h可以明显降低线粒体氧化磷酸化蛋白(NDUFS1、SDHA、UQCRC2、COXIV、OSCP)的表达,引起线粒体活性氧(ROS)产生明显增高,从而诱导细胞凋亡.结论 10058-F4可以通过降低线粒体氧化磷酸化蛋白的表达引起ROS产生来诱导CaSki细胞凋亡.     

Ⅱ型谷氨酰胺转氨酶对骨肉瘤MG-63细胞凋亡的抑制作用及其机制

目的：研究骨肉瘤MG-63细胞中Ⅱ型谷氨酰胺转氨酶（TG2）的抗凋亡作用,探讨其抑制肿瘤细胞凋亡的机制。方法：设计针对TG2的siRNA,建立骨肉瘤MG-63细胞体外缺氧培养模型,分为常氧组（细胞在常氧下培养）、单纯缺氧组（细胞在缺氧培养箱里培养）、对照siRNA缺氧组（转染对照siRNA后在缺氧培养箱里培养）和TG2 siRNA缺氧组（转染TG2 siRNA后在缺氧培养箱里培养）。观察各组骨肉瘤MG-63细胞在缺氧培养不同时间(6、12、24、48和72 h) 后Bax和Cyt C的表达及细胞凋亡率。微量滴定法检测TG2活性,RT-PCR法检测TG2和Bax mRNA表达水平,免疫组织化学法检测TG2和Bax蛋白在细胞内的分布,Western blotting法检测TG2、Bax和Cyt C蛋白表达水平,流式细胞术检测细胞凋亡率。结果：与常氧组比较,单纯缺氧组和对照siRNA缺氧组细胞TG2活性、TG2 mRNA和蛋白表达水平明显增强(P<0.01),并随缺氧时间延长逐渐升高（P<0.01）;Bax mRNA表达水平变化不明显(P>0.05),但Bax蛋白表达水平明显下降(P<0.01),细胞胞浆和线粒体内Cyt C蛋白水平及细胞凋亡率无明显变化（P>0.05）。与单纯缺氧组和对照siRNA缺氧组比较,TG2 siRNA缺氧组细胞各时间点TG2 mRNA和蛋白表达水平明显降低(P<0.01),Bax mRNA表达水平无明显变化（P>0.05）,Bax蛋白表达明显增加(P<0.01),胞浆内Cyt C蛋白水平明显增加(P<0.01),线粒体内Cyt C蛋白水平明显降低（P<0.01）,细胞凋亡率显著增加(P<0.01)。结论： TG2通过与Bax形成混合物而消耗Bax,阻止Cyt C释放至胞浆,从而抑制肿瘤细胞凋亡。     

TRAIL诱导人肝星状细胞系LX-2凋亡及DR5与Bax调节作用

目的 探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导肝星状细胞凋亡情况及DR5与Bax的调控作用.方法 RT-PCR检测LX-2中α-SMAmRNA和DR5mRNA的表达;MTT比色法、流式细胞术检测外源性TRAIL对LX-2细胞增殖和诱导细胞凋亡的影响;采用Western blot检测Bax、Caspase-3表达.结果 培养的LX-2表达αSMA mRNA和DR5mRNA逐渐增加,TRAIL可抑制其细胞增殖.TRAIL与对照组比较可以诱导活化的LX-2细胞凋亡明显增加(P<0.05),在TRAIL作用下,LX-2线粒体Bax、细胞浆Caspase-3表达上调.结论 外源性TRAIL可以诱导活化的LX-2凋亡,DR5及线粒体Bax表达上调与此过程密切相关.     

昆明山海棠碱通过线粒体途径诱导Jurkat淋巴瘤细胞株细胞凋亡

目的 研究昆明山海棠碱通过线粒体途径诱导细胞凋亡的作用。方法 THH碱多剂量多时间点诱导Jurkat细胞后，采用Western blotting检测细胞内Bcl-2表达的含量、采用JC—1原位染色方法、细胞免疫化学和流式细胞术等手段在单细胞水平分析细胞线粒体膜Apo2．7分子以及线粒体膜电位(△Ψm)的变化。结果 THH碱诱导后Bcl-2蛋白表达下调，线粒体膜电位明显下降，线粒体膜Apo2．7蛋白分子表达显著增加。结论 THH碱可通过线粒体途径诱导Jurkat T淋巴瘤细胞株发生凋亡，并且Bcl—2蛋白参与了线粒体凋亡功能的启动。     

庆大霉素损伤后内耳毛细胞凋亡与Bcl-2基因蛋白表达

目的 :探讨庆大霉素损伤时毛细胞凋亡现象及凋亡调控机制。方法 :40只豚鼠分两组 ,分别腹腔注射庆大霉素和生理盐水。利用 TUNEL标记技术和免疫组化方法分别检测毛细胞凋亡和 Bcl- 2蛋白表达。结果 :庆大霉毒实验组毛细胞凋亡发生率为 52 .6% ,Bcl- 2阳性细胞表达率为 ( 2 8.30± 4.2 4 ) % ;对照组无毛细胞凋亡现象 ,其 Bcl- 2阳性细胞表达率为 ( 56.40± 5.43) %。实验组 Bcl- 2阳性细胞表达率明显低于对照组( P<0 .0 0 1 )。结论 :1细胞凋亡是庆大霉素损伤内耳毛细胞的一种方式。 2庆大霉素可能影响凋亡相关基因的表达。     

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.