脂联素抑制晚期糖基化终末产物诱导的HUVEC-12细胞CD40高表达

目的 研究脂联素(APN)对晚期糖基化终末产物(AGEs)诱导HUVEC-12细胞CD40高表达的干预作用.方法 ①用不同浓度AGE(0、100、200、400和800 mg/L)处理细胞24 h,采用实时定量PCR和Western Blot技术分别检测HUVEC-12细胞CD40 mRNA和蛋白质的表达.②用不同浓度APN(0、5、10和15μg/mL)预处理细胞,加入AGEs(400mg/L)共同培养细胞24 h,采用实时定量PCR和Western Blot技术检测HUVEC-12细胞CD40mRNA和蛋白质的表达.结果 ①不同浓度AGEs处理细胞24 h,随着处理浓度增大,HUVEC-12细胞CD40的表达逐渐上调;②不同浓度APN预处理细胞后,随着浓度增大,APN逐渐下调AGEs诱导的HUVEC-12细胞CD40高表达,对正常状态下的CD40表达没有影响.结论 AGEs诱导HUVEC-12细胞CD40表达上调,脂联素具有抑制AGEs诱导HUVEC-12细胞CD40高表达的作用.     

DFMG对TNF-α诱导的人脐静脉内皮细胞CD40/CD40L相互作用的影响

目的:观察7-二氟甲氧基-5,4’-二甲氧基金雀异黄素(DFMG)对肿瘤坏死因子-α(TNF-α)诱导人脐静脉内皮细胞(HUVE-12)损伤模型的保护作用,探讨其保护作用的发挥是否与抑制CD40/CD40L相互作用相关。方法:体外培养人脐静脉内皮细胞(HUVE-12),20ng/mL TNF-α孵育诱导细胞损伤模型;加入不同浓度的DFMG,台盼蓝拒染法检测细胞活力、AnnexinⅤ-FITC/PI双染色流式细胞术检测细胞凋亡、流式细胞仪检测CD40/CD40L的表达。结果:DFMG呈浓度依赖性的升高TNF-α诱导的HUVEC-12细胞存活率;DFMG呈浓度依赖性的降低TNF-α诱导的HUVEC-12细胞凋亡;DFMG呈浓度依赖性降低CD40/CD40L表达。结论:DFMG保护TNF-α诱导的HUVEC-12细胞损伤,这种保护作用可能是通过抑制CD40/CD40L的表达发挥作用的。     

晚期糖基化终末产物对巨噬细胞Nampt表达的影响

目的 探讨晚期糖基化终末产物(AGEs)对巨噬细胞尼克酰胺磷酸核糖转移酶(Nampt)表达的影响.方法 以人单核细胞株THP-1为体外细胞干预模型,经佛波酯诱导分化为巨噬细胞后,以0(阴性对照组)、50、100、250、500、1 000μg/mL的AGEs干预12 h,采用Western blotting方法检测细胞Nampt蛋白表达.以Western blotting检测结果选择合适浓度的AGEs处理巨噬细胞,分别于干预后0(阴性对照组)、3、6、12、24 h时点收集细胞,采用实时定量聚合酶链式反应(Real-Time PCR)技术和Western blotting方法检测巨噬细胞Nampt mRNA和蛋白表达.结果 在50～500μg/mL的浓度范围内,AGEs处理组巨噬细胞Nampt蛋白表达量均显著高于阴性对照组(P＜0.01),且随干预浓度的升高呈现逐渐上升趋势,250μg/mL和500μg/mL AGEs处理组的干预效应最显著,设定后续实验中以250μg/mL AGEs对巨噬细胞进行干预.250μg/mL AGEs处理组巨噬细胞干预后各时点的Nampt mRNA和蛋白表达量均显著高于阴性对照组(P＜0.01),且随干预时间的延长呈现逐渐上升趋势.结论 AGEs干预能促进巨噬细胞Nampt表达上调;提示AGEs 可能部分通过Nampt途径激活巨噬细胞而参与糖尿病动脉粥样硬化的发生过程.     

高糖、糖基化终产物对大鼠主动脉平滑肌细胞巨噬细胞炎性蛋白1α表达的影响

目的:探讨高糖浓度和糖基化终产物(AGEs)对大鼠主动脉平滑肌细胞巨噬细胞炎性蛋白(MIP)1αmRNA及蛋白表达的影响.方法:将培养的大鼠主动脉平滑肌细胞分别用不同浓度AGEs(100,200,400 mg/L)单独培养24h;400mg/L AGEs和22mmol/L葡萄糖联合培养24 h.RT-PCR方法及流式细胞仪检测MIP-1α mRNA及蛋白的表达.结果:对照组平滑肌细胞内MIP-1α呈弱表达;100,200,400 mg/L AGEs培养平滑肌细胞24 h及22mmol/L葡萄糖和400 mg/L AGEs联合孵育24 h后显著增高MIP-1α mRNA的表达,其电泳条带相对积分吸光度值分别为对照组的1.36,1.75,2.45及2.76倍(P＜0.05);各组平滑肌细胞MIP-1α蛋白的平均荧光强度分别为对照组的1.24,1.63,2.37及2.68倍(P＜0.05).结论:AGEs能刺激平滑肌细胞MIP-1α mRNA及蛋白表达增加,且与高糖具有协同作用.     

糖基化终产物AGEs对U937巨噬细胞高密度脂蛋白受体(SR-BI)蛋白表达的影响

目的研究诱导分化及糖基化终产物对人单核/巨噬细胞(U937细胞)高密度脂蛋白受体(SR-BI)蛋白表达的影响.方法U937细胞经PMA诱导分化,并将不同浓度或同一浓度AGEs与诱导分化48h后的U937细胞共同孵育,用免疫细胞化学法和Western印迹法检测SR-BI蛋白的表达.结果诱导分化后U937细胞SR-BI表达在24、48 h逐渐升高,72 h下降;100、200和400mg/L AGEs刺激后细胞表面SR-BI蛋白表达量分别是BSA组的1.44、2.38和2.77倍(P＜0.05);400mg/L的AGEs作用6、12、24、48 h后,U937巨噬细胞SR-BI蛋白表达量分别为0 h的1.38、2.49、3.76和4.25倍(P＜0.05).结论诱导分化24及48 h SR-BI蛋白表达逐渐增加,而分化72h蛋白表达下降;AGEs可增加U937巨噬细胞SR-BI蛋白表达且呈浓度和时间依赖性.     

球状脂联素抑制晚期糖基化终产物诱导人脐静脉内皮细胞凋亡的机制研究

目的 观察球状脂联素(globular adiponectin,gAd)是否抑制晚期糖基化终产物(advanced glycation end products,AGEs)诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)凋亡,并探讨其相关的机制.方法 用不同浓度(100、200和400 mg/L)AGEs培养HUVECs 24 h或48 h,其中一组预先用3mg/L gAd处理24 h.MTT比色法测定细胞活性,Annexin V-FITC/PI双染标记流式细胞仪检测细胞凋亡,免疫印记检测phospho一α AMPK和αAMPK蛋白表达,以及TT-PCP,检测内皮细胞脂联素受体1(adiponectin receptor 1,AdipoR1)mRNA的表达.结果 与对照组比较,随着AGEs浓度的增加,细胞活性显著下降(P<0.01),细胞凋亡率显著增加(P<0.01).与200mg/L AGEs组比较,经gAd预处理后的200mg/L AGEs组细胞活性显著增加(P<0.01),细胞凋亡率显著下降(P<0.01),P-αAMPK/αAMPK蛋白表达显著增加(P<0.01),AdipoR1 mRNA表达显著增加(P<0.01).结论 gAd可能通过激活AMPK/AdipoR1通路抑制AGEs诱导HUVECs发生凋亡.     

晚期糖基化终产物对内皮细胞Connexin40基因表达的影响

目的 研究晚期糖基化终产物(advanced glycation end products,AGEs)对体外培养的人脐静脉内皮细胞缝隙连接蛋白Connexin40 基因表达的影响.方法 D- 葡萄糖孵育牛血清白蛋白制备糖基化牛血清白蛋白作为晚期糖基化终产物.体外常规培养的人脐静脉内皮细胞分为4组,分别为对照组、100mg/L AGEs 干预组、200mg/L AGEs 干预组和400mg/L AGEs 干预组,干预24h 后通过RT-PCR 方法检测各组Connexin40 基因mRNA 的表达有无差异;通过免疫细胞荧光方法检测对照组和200mg/LAGEs 干预组之间Connexin40 基因蛋白的表达水平有无差异.结果 不同浓度(100、200、400mg/L)的AGEs 干预24h 均能引起人脐静脉内皮细胞Connexin40 基因mRNA 的表达减少(均P＜0.01);200mg/L 的AGEs 干预24h 引起人脐静脉内皮细胞Connexin40基因的蛋白表达减少(P＜0.01).结论 AGEs 体外短期干预可引起人脐静脉内皮细胞Connexin40 基因的mRNA 和蛋白表达水平减少.     

晚期糖基化终产物引起内皮细胞connexin43表达上调

目的研究晚期糖基化终产物(advanced glycation end products,AGEs)对体外培养的人脐静脉内皮细胞缝隙连接蛋白(gap junction protein)connexin43基因表达的影响。方法 D-葡萄糖孵育牛血清白蛋白制备糖基化牛血清白蛋白作为晚期糖基化终产物。体外常规培养的人脐静脉内皮细胞分为4组,分别为对照组,100、200和400mg/L AGEs干预组,干预24h后通过RT-PCR方法检测各组connexin43基因mRNA的表达有无差别;通过免疫荧光方法检测对照组和200mg/L AGEs干预组之间connexin43蛋白的表达水平有无差别。结果不同浓度(100、200、400mg/L)的AGEs干预24h均能引起人脐静脉内皮细胞con-nexin43基因mRNA的表达上调(P<0.01);200mg/L的AGEs干预24h引起人脐静脉内皮细胞connexin43基因的蛋白表达上调(P<0.01)。结论 AGEs体外短期干预可引起人脐静脉内皮细胞connexin43基因的mRNA和蛋白表达水平上调。     

晚期糖基化终末产物对巨噬细胞Nampt表达的影响

目的探讨晚期糖基化终末产物(AGEs)对巨噬细胞尼克酰胺磷酸核糖转移酶(Nampt)表达的影响。方法以人单核细胞株THP-1为体外细胞干预模型,经佛波酯诱导分化为巨噬细胞后,以0(阴性对照组)、50、100、250、500、1 000μg/mL的AGEs干预12 h,采用Western blotting方法检测细胞Nampt蛋白表达。以Western blotting检测结果选择合适浓度的AGEs处理巨噬细胞,分别于干预后0(阴性对照组)、3、6、12、24 h时点收集细胞,采用实时定量聚合酶链式反应(Real-Time PCR)技术和Westernblotting方法检测巨噬细胞Nampt mRNA和蛋白表达。结果在50～500μg/mL的浓度范围内,AGEs处理组巨噬细胞Nampt蛋白表达量均显著高于阴性对照组(P<0.01),且随干预浓度的升高呈现逐渐上升趋势,250μg/mL和500μg/mL AGEs处理组的干预效应最显著,设定后续实验中以250μg/mL AGEs对巨噬细胞进行干预。250μg/mL AGEs处理组巨噬细胞干预后各时点的Nampt mRNA和蛋白表达量均显著高于阴性对照组(P<0....     

AGEs诱导软骨细胞凋亡和影响细胞外基质变化的研究

目的 :明确晚期糖基化终末产物(advanced glycation end products,AGEs)诱导小鼠关节软骨细胞凋亡情况和影响细胞外基质变化。方法:体外分离培养小鼠软骨关节细胞,给予不同浓度AGEs共培养后,分别于24 h和48 h时间点观察细胞凋亡情况;通过Western blot方法检测Ⅰ型胶原、MMP-3、MMP-9以及p53蛋白的表达情况。结果:不同浓度AGEs处理软骨细胞24 h后,与对照组比较,细胞凋亡率出现升高(P<0.05,n=3);与BSA组比较,200 mg/L浓度的AGEs组细胞凋亡率明显升高(P<0.05,n=3),并且随着AGEs浓度和培养时间增加,软骨细胞的凋亡率也增加。200 mg/L的浓度AGEs处理软骨细胞48 h后,Ⅰ型胶原蛋白的表达显著下降(P<0.05,n=3);另外,50、100、200 mg/L的浓度AGEs处理软骨细胞48 h后,p53、MMP-3、MMP-9的表达显著高于对照组和BSA组(P<0.05,n=3)。结论 :AGEs能诱导小鼠软骨细胞凋亡,并刺激软骨细胞p53、MMP-3、MMP-9表达增多,降低Ⅰ型胶原蛋白表达。     

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.     

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.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.