赖氨酰氧化酶对胃癌细胞血管生成拟态的影响

目的 研究赖氨酰氧化酶（lysyl oxidase,LOX）对胃癌细胞MGC-803和BGC-823体外血管生成拟态（vasculogenic mimicry,VM）的影响.方法 培养胃癌细胞,分别加入含有不同浓度LOX或LOX的抑制剂β-氨基丙腈（beta-aminopropionitrile,BAPN）作用72h,用过碘酸雪夫（periodic acid-schiff stain,PAS）染色方法检测VM形成的数量.结果 在两种胃癌细胞株中,随着LOX浓度的增加,VM形成的数量也增加,LOX浓度与VM的形成呈正相关（r=0.847和r=0.851）;此效应可被BAPN抑制,随着BAPN的浓度增加,VM形成的数量相应减少,BPAN与VM的形成呈现负相关（r=-0.791和r=-0.834）.结论 外源性添加LOX可以促进胃癌MGC-803和BGC-823细胞株VM的形成.     

转化生长因子β1对内皮细胞屏障功能的影响

目的观察转化生长因子β1（TGF-β1）对体外培养人脐静脉内皮细胞株EA.hy926屏障功能的影响并探讨其作用机制。方法体外培养EA.hy926脐静脉内皮细胞株,实验分为两组,对照组培养液中加入PBS;TGF-β1处理组培养液中添加0.01mg/L的TGF-β1。将细胞在多聚脂滤膜上培养,测量细胞跨上皮电阻（TER）的变化。免疫荧光检测两组血管内皮钙黏蛋白（VE-cadherin）、α-连环蛋白、血管内皮细胞生长因子（VEGF）和纤维连接蛋白的表达变化;RT-PCR检测两组VE-cadherin和VEGF表达的变化。结果TGF-β1处理后TER降低,两组处理前后TER差值比较有统计学意义（t=3.294,P〈0.01）;TGF-β1处理组VE-cadherin和α-连环蛋白表达减少,内皮细胞黏附连接被破坏;而VEGF和纤维连接蛋白表达增多,基底膜增厚。结论TGF-β1可引起内皮细胞屏障功能的破坏,其机制可能与VEGF的表达上调有关。     

β-氨基丙腈抑制赖氨酰氧化酶对小鼠免疫功能的影响

目的观察β-氨基丙腈(β-Aminopropionitrile,BAPN)抑制赖氨酰氧化酶(lysyl oxidase,LOX)后小鼠免疫功能的变化。方法选用18~22g的正常ICR小鼠,以β-氨基丙腈100mg/kg连续灌胃10d。处死后检测小鼠胸腺重、脾重、巨噬细胞吞噬功能;采用绵羊红细胞作为抗原,检测小鼠抗体生成细胞功能;采用MTT法检测小鼠脾淋巴细胞对ConA刺激的转化能力。结果以β-氨基丙腈抑制LOX活性,明显降低小鼠胸腺/体重比值;巨噬细胞吞噬率及吞噬指数均下降;小鼠抗体生成细胞数减少;ConA诱导的小鼠脾淋巴细胞增殖能力减弱。结论β-氨基丙腈抑制赖氨酰氧化酶可减弱小鼠免疫功能。     

高糖诱导血管内皮细胞表达结缔组织生长因子的影响

目的观察高糖对血管内皮细胞表达结缔组织生长因子（connective tissue growth faeotr, CTGF）的影响。方法体外培养人脐静脉内皮细胞（human umibilical vein endotheial cells,HUVECs）,分别在含正常葡萄糖（5．5mM）、高糖（25mM葡萄糖）、25mM甘露醇及高糖加转化生长因子β1（transforming frowth factorβ1,TGFβ1）中和抗体的培养基中培养24、48或72小时,并在相应时间点收集细胞总RNA及总蛋白。分别用半定量RT-PCR及Western Blot方法检测CTGFmRNA及蛋白表达水平的变化。结果与正常葡萄糖培养及25mM甘露醇对照组相比,高糖刺激24小时后,HUVECs的CTGFmRNA及蛋白表达水平均显著增加,并且这种作用持续到48小时。TGFβ1的中和抗体能部分阻断高糖的这种刺激作用。结论高糖能诱导HUVECs表达CTGF,而内皮细胞CTGF的表达增加可能是高糖参与动脉粥样硬化等心血管疾病发生的机制之一。     

健心颗粒对血管内皮细胞KLF4表达的影响

目的观察健心颗粒对人脐静脉血管内皮细胞炎症损伤的干预作用。方法通过炎症因子TNF—α诱导人脐静脉内皮细胞建立内皮细胞损伤模型,随机分为正常组、健心颗粒组、TNF-α组、TNF-α加健心颗粒组,用RT—PCR、Western—Blot、免疫组化法检测脐静脉内皮细胞Kruppel因子4（KLF4）表达的变化。结果与正常组比较,TNF-α刺激后,KLF4表达水平明显升高;健心颗粒干预后,KLF4表达水平下降。结论健心颗粒能够抑制激活的人脐静脉血管内皮细胞中KLF4的表达。     

体外温热刺激对脐静脉血管内皮细胞线粒体及三磷酸腺苷含量影响的实验研究

目的：研究体外适宜温度刺激对人脐静脉血管内皮细胞线粒体及三磷酸腺苷（ATP）含量变化的影响,探讨灸法中温热刺激温阳作用的能量转换机制.方法：选取原代人脐静脉血管内皮细胞进行体外培养,分为体外适宜温度刺激组（40℃）和正常组（37℃）,体外适宜温度刺激组40℃持续刺激30min,每天2次,共刺激3天;第3天刺激结束4～6小时后用詹纳斯绿B活染法观察细胞线粒体变化,高效液相色谱法检测人脐静脉血管内皮细胞ATP含量.结果：适宜温度40℃组与正常组37℃比,人脐静脉血管内皮细胞线粒体数量增多,细胞内ATP含量升高（p＜0.01）.结论：体外适宜温度刺激后能够促进人脐静脉血管内皮细胞的线粒体增长,提高脐静脉血管内皮细胞代谢的能量物质,从而为灸法的温阳作用提供了实验依据.     

血管内皮生长因子对缺氧诱导内皮细胞凋亡的影响

目的 探讨缺氧对培养人静脉内皮细胞凋亡的影响及血管内皮生长因子的干预性影响。方法 （1）人脐静脉内皮细胞的培养及鉴定。（2）建立人脐静脉内皮细胞缺氧模型,TUNEL法观测不同缺氧时间（0、12、24、48h）对内皮细胞凋亡的影响及血管内皮生长因子的干预作用。结果 随缺氧时间延长,NUVECs凋亡率显著升高,血管内皮生长因子抑制缺氧导致的内皮细胞凋亡。结论 内皮细胞的过渡凋亡是引起内皮功能障碍的一个重要因素,血管内皮生长因子通过抑制内皮细胞凋亡,而具有内皮细胞保护作用。     

高糖刺激人脐静脉内皮细胞丝裂素活化蛋白激酶P38的表达

目的：探讨丝裂素活化蛋白激酶P38（P38MAPK）在糖尿病血管并发症中的作用。方法：采用体外培养和Western-blot等方法，以不同浓度D－葡萄糖及不同时间刺人脐静脉内皮细胞系，分别检测人脐静脉内皮细胞系P38MAPK的蛋白表达。结果：随D－葡萄糖浓度增加，P38MAPK磷酸化蛋白表达量增加；延长刺激时间，P38MAPK磷酸化蛋白表达量亦随之增加。结论：高糖引发的P38MAPK表达异常，可能在糖尿病血管并发症的发生、发展中起关键作用。     

抑肽酶对内皮细胞β-链蛋白保护作用的实验研究

目的体外观察人肿瘤坏因子α（TNFα）对内皮细胞细胞连接蛋白β-链蛋白的影响及抑肽酶的保护作用，探讨抑肽酶抑制体外循环术后系统性炎性反应的机制。方法将体外培养的人脐静脉内皮细胞随机分为空白组，TNFα组，抑肽酶＋TNFα组。采用免疫细胞化学法观察人脐静脉内皮细胞连接蛋白β-链蛋白（β-catenin）的表达。用细胞培养小室测定中性粒细胞跨迁单层融合的人脐静脉内皮细胞的迁移率。结果250KIU／ml抑肽酶可显著减少β—catenin的丢失（P〈0．05）；250KIU／ml抑肽酶对中性粒细胞跨迁活化的单层人脐静脉内皮细胞有显著影响（P〈0．05）。结论抑肽酶对内皮细胞细胞连接具有保护作用；抑肽酶可能通过保护内皮细胞细胞连接来抑制体外循环术后系统性炎性反应。     

靶向siRNA沉默Akt1基因表达及其对肺癌培养基促内皮细胞迁移的抑制作用

目的:研究小干扰RNA(siRNA)对血管内皮细胞Akt1基因表达的沉默作用及其对肺癌A549细胞条件培养基促血管内皮细胞迁移的影响。方法:以人脐静脉内皮细胞为靶细胞,设计针对Akt1基因mRNA的siRNA(siAkt1),利用阳离子脂质体介导转染人脐静脉内皮细胞,采用半定量PCR技术(RT-PCR)检测Akt1mRNA的表达,用Western blotting技术检测Akt总蛋白表达。同时用体外"伤口愈合"实验观察细胞迁移。结果:筛选出的siAkt1能显著抑制内皮细胞Akt1mRNA(P<0.01)和Akt总蛋白的表达(P<0.05),A549细胞条件培养基显著促进人脐静脉内皮细胞迁移(P<0.05),但对转染siAkt1的人脐静脉内皮细胞迁移没有促进作用(P>0.05)。结论:特异性siRNA通过有效抑制Akt1基因的表达而抑制A549细胞条件培养基促血管内皮细胞迁移的作用,这为肺癌血管生成干预治疗提供了新的理论基础。     

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.     

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

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.