迷迭香精油诱导宫颈癌Hela细胞凋亡的实验研究

【目的】研究迷迭香精油对体外培养Hela细胞生长的抑制作用。【方法】采用水蒸气蒸馏法提取迷迭香精油,气相色谱-质谱联用仪分析成份。以不同浓度的迷迭香精油作用于体外培养的Hela细胞,MTT法测定迷迭香精油抗肿瘤活性,透射电镜观察药物对细胞形态学的影响。流式细胞仪检测细胞凋亡率。【结果】MTT法证明迷迭香精油可显著抑制Hela细胞生长,抑制率与药物浓度正相关,呈现明显的量-效和时-效关系,细胞经药物作用48h后的IC50为0.0736mg/L,电镜下可观察到核浓缩及核碎裂等典型的细胞凋亡特征。流式细胞仪可以检测到细胞凋亡。【结论】迷迭香精油可抑制Hela细胞的生长,诱导Hela细胞凋亡。     

融合蛋白XE-TNFαm2对SIV的抑制作用

目的检测XE-TNFαm2靶向融合蛋白作为新型生物性药物抑制SIV病毒感染CEMx-174细胞的作用。方法 XE-TNFαm2、SIV和CEMx-174细胞共同培养,在培养的第5天,用酶联免疫吸附法(ELISA)测定培养上清中P27抗原水平,间接免疫荧光法(IFA)和流式细胞仪测试SIV阳性细胞,应用PE-annexin V凋亡检测试剂盒测试细胞存活率和凋亡率。结果 XE-TNFαm2在细胞安全最大药物浓度50μg/mL时,对SIV病毒抑制率为40%。细胞早期凋亡率在XE-TNFαm2浓度6.25μg/mL时为37.1%。结论融合蛋白XE-TNFαm2对SIV病毒有一定的抑制作用,其中部分作用推测是因通过诱导感染细胞产生凋亡导致病毒死亡。     

卷柏多糖对肠道病毒71型复制的体外抑制作用

目的研究卷柏多糖对EV71病毒复制的体外抑制活性。方法将卷柏多糖不同提取物设置不同剂量作用于人恶性横纹肌肉瘤细胞(RD),采用细胞病变效应(CPE)及细胞计数试剂盒(CCK8法)检测细胞存活率,并计算药物的半数中毒浓度(CC50);培养EV71感染的RD细胞,建立体外病毒感染模型,并设立正常细胞对照组和阳性药物对照组(利巴韦林3.2mg/L),将卷柏多糖不同提取物设置不同剂量作用于EV71感染后的RD细胞,应用CCK8法检测药物抗病毒抑制率,并计算药物的半数抑制浓度(IC50);收集药物作用于感染后RD细胞的培养液上清,进行荧光定量PCR检测,观察药物对EV71病毒RNA表达的影响。结果卷柏30%醇沉多糖和50%醇沉多糖对RD细胞的CC50值分别为396和142 mg/L,抑制EV71病毒致CPE的IC50值分别为40.8和26.2 mg/L。与病毒感染对照组相比较,卷柏30%和50%醇沉多糖作用后,经EV71感染的RD细胞培养液中病毒RNA拷贝数显著降低(P<0.05)。结论卷柏及其30%和50%醇沉多糖可以作为新的抗EV71的潜在药物。     

双氢青蒿素抗恶性脑胶质瘤作用的研究

目的 研究双氢青蒿素对人恶性脑胶质瘤U87细胞生长的作用. 方法 培养人恶性脑胶质瘤U87细胞,MTT法检测双氢青蒿素作用后U87 细胞的生长抑制率,倒置显微镜观察药物作用后细胞生长的变化.结果 0.5、5.0、50 μmol/L双氢青蒿素均明显抑制U87细胞的生长,倒置显微镜下观察到药物作用后细胞密度明显降低. 结论 双氢青蒿素能明显抑制脑胶质瘤细胞的生长增殖.     

31例肺癌细胞药敏试验及其临床应用

目的　了解肿瘤细胞药敏试验在肺癌化疗中的临床意义。方法　取肺癌细胞进行琼脂培养 ,用MTT法进行药敏测定。根据药敏结果选择药物治疗肺癌。结果　对MCC、5 Fu的敏感性 >50 % ,阳性符合率达 80 % ,阴性符合率为 81.2 %。结论　根据药敏结果选择药物治疗肺癌临床符合率高 ,疗效满意。     

黄芪甲甙体外抗乙型肝炎病毒的作用

目的：探讨黄芪甲甙对乙型肝炎病毒（HBV）的体外抑制作用．方法：以HepG2．2．15细胞株为细胞模型，分别加入不同浓度的黄芪甲甙和拉米夫定，于培养第3，6和9日收集培养上清液．通过MqT法观察药物对HepG2．2．15细胞株的影响，采用ELISA法及荧光定量PCR法，分析药物对细胞培养上清液中HBsAg，HBeAg及HBVDNA含量的影响．结果：黄芪甲甙具有一定程度的抗HBV的作用．黄芪甲甙给药9d，其HBsAg的50％抑制浓度（Ic50）为22．1mg／L，治疗指数（TI）为4．4；HBeAg的IC50为26．2mg／L，TI为3．72；细胞外HBVDNA的IC50为13．2mg／L，TI为7．4．拉米夫定也有抑制HBV的作用，但抑制作用较弱．结论：黄芪甲甙体外对HBV有抑制作用．     

MTT法检测乳腺癌原代细胞中SP细胞对化疗药物的敏感性

目的:探讨MTT法在筛选乳腺癌较敏感的药物及浓度和指导乳腺癌临床治疗中的作用.方法:乳腺癌原代细胞和乳腺癌细胞株MCF-7培养传代,利用Hoechest DNA染色流式细胞仪分选SP细胞和非SP细胞.选SP细胞、MP细胞、PP细胞分别加入不同浓度的四种常规化疗药物观测SP细胞对药物敏感性.结果:不同药物对SP、MP和PP三种细胞的半抑制浓度IC50都呈现如下趋势:SP>>PP>MP.结论:SP细胞对不同药物都具有最低的敏感性,会将药物之间的差异放大,更适合用其检测药物毒性,药物毒性随着药物浓度的增加而增高,在具有相同效应的药物中,应选择低浓度,但因药物的作用机理不尽相同,患者身体基础状况及耐受程度不同,还应结合临床试验确定药物浓度.     

人脑胶质瘤原代细胞培养与药敏实验

我们以组织块法作了43例人脑胶质瘤细胞原代培养。存活生长39例(92.5%),其中32例完成了平阳霉素,卡氮芥(BCNU)、长春新碱(VCR)各浓度的药敏测定。以细胞生长状况与形态学改变为指标.观察各个体胶质瘤培养细胞对不同药物的敏感性。在各药理论有效浓度下的敏感率:平阳霉素为78.1%、BCNU 为75%、VCR 为50%。本实验方法简便,成功率高。接种后平均8天开始生长,21天可作药敏试验,29天能获知结果。对临床脑胶质瘤化疗药物的选用不失为一个有用的客观指标。     

淫羊藿苷诱导小鼠胚胎干细胞体外定向分化为心肌细胞方法学研究

目的 采用药物淫羊藿苷(icariin,ICA)改变小鼠胚胎干细胞(embryonic stem cell,ES细胞)体外培养的微环境,建立一种药物介导提高ES细胞体外定向心肌细胞分化率的实验体系.方法 采用悬滴培养法,构建ICA诱导ES细胞体外定向分化为心肌细胞的实验评价体系;并利用RT-PCR法和免疫荧光法鉴定心肌细胞.结果 ICA在10-7mol&#183;L-1浓度时,对ES细胞定向分化为心肌细胞呈现最佳诱导效应,分化率最高达84%(P＜0.001).ES细胞源心肌细胞表达心肌特异性基因肌球蛋白重链(α-MHC)和心室肌球蛋白轻链(MLC-2v),且心肌特异性肌小节α辅肌动蛋白(α-actinin)和肌钙蛋白T(troponin T)呈阳性表达.结论 悬滴培养法结合药物改善微环境,可用于药物诱导ES细胞定向分化心肌细胞的实验体系.     

光化学法病毒灭活的模型病毒培养与滴定方法的建立

目的建立Sindbis和伪狂犬病毒的培养与滴定方法.方法以Vero细胞为病毒传代及滴度滴定指示细胞,Karher氏法计算滴定结果.结果 Sindbis病毒感染的Vero细胞以圆缩、脱落为主;伪狂犬病毒感染的Vero细胞以变圆、形成折光性强的合胞体为主,病变细胞晚期脱落.Sindbis病毒第一次滴定TCID50＞11,第二次滴度TCID50为9.5;伪狂犬病毒第一次滴定TCID50＞11,第二次滴定TCID50为7.2.结论 Vero细胞对两种模型病毒均较敏感,适合用于此两种病毒的培养与滴定.该方法的建立为光化学法灭活血小板中病毒的研究奠定基础.     

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.