RNA干扰沉默乙酰肝素酶基因对卵巢癌微血管内皮细胞的影响

目的 探讨RNA干扰沉默乙酰肝素酶基因对卵巢癌微血管内皮细胞的影响.方法 实验组为乙酰肝素酶 siRNA转染组(转染组),实验对照为正常组和siRNA空白转染对照组(空白转染组),观察时间为转染后2、4、6、8d .观察血管内皮细胞乙酰肝素酶 siRNA转染效率,用MTT法检测细胞活力,流式细胞仪检测细胞周期变化,免疫荧光、RT-PCR法及WB法检测内皮细胞乙酰肝素酶表达.结果 转染组细胞生长受抑制,增殖指数显著下降,乙酰肝素酶mRNA水平和蛋白水平显著下降,细胞凋亡率显著增加.结论 RNAi可有效沉默卵巢癌微血管内皮细胞乙酰肝素酶表达,提示RNAi可为卵巢癌治疗提供新的途径.     

乙酰肝素酶和syndecan-1表达与脑胶质瘤恶性程度的关系

目的:检测脑胶质瘤组织中乙酰肝素酶和syndecan-1的表达水平,探讨它们和肿瘤级别的关系及作用机制。方法:采用荧光实时定量聚合酶链式反应(RT-PCR)法检测乙酰肝素酶mRNA和syndecan-1 mRNA在40例脑胶质瘤组织和10例良性脑肿瘤组织中的表达水平,统计分析表达水平和肿瘤级别之间的关系。结果:在胶质瘤中,乙酰肝素酶的表达明显高于良性脑肿瘤(P<0.01),并且随着肿瘤的恶性程度增高,乙酰肝素酶的表达也明显增高(P<0.01),syndecan-1的表达明显低于良性脑肿瘤(P<0.01),并且随着肿瘤的恶性程度增高,syndecan-1的表达也明显降低(P<0.01)。乙酰肝素酶与syndecan-1在Ⅰ~Ⅱ级脑胶质瘤中表达均呈负相关关系(P<0.01),在Ⅲ~Ⅳ级脑胶质瘤中表达均呈负相关关系(P<0.01),在各级胶质瘤中表达相关性的总体分析均显示负相关关系(P<0.01)。结论:乙酰肝素酶在脑胶质瘤组织中高表达,syndecan-1在脑胶质瘤中低表达,而且表达水平和恶性程度密切相关,它们的检测可以作为脑胶质瘤恶性程度判断的参考,为探讨脑胶质瘤的生物学行为、预后及治疗提供新的思路。     

北极刺参多糖、皂苷和胶原蛋白多肽对血管内皮细胞的保护作用

目的:研究北极刺参多糖、皂苷和胶原蛋白多肽对血管内皮细胞的保护作用。方法:采用氧化型低密度脂蛋白(ox-LDL)建立体外培养的人脐静脉血管内皮细胞株(ECV304)损伤模型。MTT法测定血管内皮细胞的增殖活性,硝酸还原酶法测定血管内皮细胞一氧化氮合酶(NOS)活力和NO释放量,TBA法测定细胞内丙二醛(MDA)含量,DNA-Ladder法检测血管内皮细胞的凋亡。结果:不同浓度的北极刺参多糖和胶原蛋白多肽均能显著抑制ox-LDL对血管内皮细胞的氧化损伤,促进血管内皮细胞增殖(P<0.05,P<0.01),降低细胞内MDA含量(P<0.05,P<0.01),促进血管内皮细胞NO释放量(P<0.05,P<0.01),提高细胞NOS活力(P<0.05,P<0.01),拮抗ox-LDL引起的血管内皮细胞凋亡。刺参皂苷具有显著的细胞毒性作用,仅能降低ox-LDL氧化损伤细胞内MDA含量(P<0.05,P<0.01),而无其他上述作用。结论:北极刺参多糖和胶原蛋白多肽对血管内皮细胞的脂质过氧化物损伤均具有显著的保护作用。     

乙酰肝素酶和CD105在胃癌中的表达及其相关性

目的:探讨乙酰肝素酶和CD105在胃癌中的表达以及它们之间的关系。方法:应用免疫组化SP方法检测乙酰肝素酶和CD105在90例胃癌组织中的表达,记数肿瘤微血管密度(microvessel density,MVD),分析乙酰肝素酶mRNA表达与胃癌浸润、转移和血管生成之间的关系。结果:90例胃癌组织中,乙酰肝素酶mRNA阳性表达46例(51.11%),MVD平均值为(71.3±18.6),阴性表达44例(48.89%),MVD平均值为(40.1±11.8)。乙酰肝素酶阳性组MVD表达与阴性组相比有显著性差异(P<0.01)。结论:乙酰肝素酶可促进胃癌的浸润、转移和血管生成,可作为反映胃癌生物学行为的客观指标。     

姜黄色素活性单体成分抗血管生成的实验研究

目的探讨中药姜黄的活性单体成分姜黄素(姜黄素Ⅰ)、脱甲氧基姜黄素(姜黄素Ⅱ)和双脱甲氧基姜黄素(姜黄素Ⅲ)对血管生成的抑制作用.方法①以体外培养的人脐静脉内皮细胞和人肺微血管内皮细胞为对象,研究3种单体对内皮细胞增殖的抑制作用和诱导内皮细胞凋亡的效果;②建立裸鼠人肺腺癌细胞A549移植瘤模型,腹腔注射双脱甲氧基姜黄素,研究其对荷癌裸鼠肿瘤血管生成的影响.结果①MTT检测3种姜黄色素作用72 h后对内皮细胞增殖抑制作用的IC50值分别为0.525、0.399、0.125 μg/ml, 在浓度为4 μg/ml时,3种姜黄色素作用48 h后均导致内皮细胞出现明显的增殖抑制(P<0.05),作用72 h时达最高值;②流式细胞仪分析浓度为4 μg/ml时姜黄素Ⅲ可将内皮细胞阻止于S期(P<0.05),浓度增至8 μg/ml以上时不但可引起S期细胞比例增加(P<0.01),同时又有明确的诱导凋亡作用;③免疫组化显示姜黄素Ⅲ实验组裸鼠瘤组织内新生血管的数量明显减少,与阴性对照组相比相差非常显著(P<0.01);姜黄素Ⅲ组肿瘤组织中VEGF表达水平,VEGF受体Flt-1、KDR水平与阴性对照组相比均相差非常显著(P<0.01).结论 3种姜黄色素单体均具有确切的抗人内皮细胞和肺腺癌裸鼠移植瘤血管生成作用,其中姜黄素Ⅲ最强,机制可能与其抑制VEGF及其受体的表达有关.     

上皮性卵巢癌中Maspin的表达及其与血管生成的关系

目的 研究丝氨酸蛋白酶抑制剂mammary serpin(Maspin)在上皮性卵巢癌(EOC)中的表达及其与血管生成的关系.方法 采用免疫组化LsAB法对31例正常卵巢组织、35例上皮性卵巢癌标本进行Maspin、VEGF、乙酰肝素酶(HPA)表达检测,CD34抗体标记血管内皮细胞计数微血管密度,分析它们与卵巢癌临床病理特点的关系.结果 Maspin、VEGF、HPA、CD34在卵巢癌中的表达与正常卵巢组织相比明显上调(P<0.05);Maspin蛋白表达与非浆液性卵巢癌、低临床分期、无腹水形成直接相关(P<0.05);VEGF蛋白表达与卵巢癌腹水形成有显著正相关性(P<0.05);HPA蛋白表达与卵巢癌淋巴结转移、腹水形成有显著正相关性(P<0.05);微血管密度与卵巢癌淋巴结转移、临床分期有显著正相关性(P<0.05);卵巢癌中Maspin的蛋白表达与VEGF(r=-0.738,P<0.05)、微血管密度有显著负相关(r=-0.655,P<0.05),与HPA无直接相关性(P>0.05);VEGF与微血管密度有正相关性(r=0.661,P<0.05);HPA与VEGF、微血管密度无直接相关性(P>0.05).结论 Maspin对上皮性卵巢癌的血管生成、侵袭、转移有一定抑制作用.     

低氧对大鼠肺动脉平滑肌细胞和肺微血管内皮细胞表达及分泌fractalkine的影响

目的 观察低氧对培养的大鼠肺动脉平滑肌细胞和肺微血管内皮细胞表达及分泌fractalkine（FKN）的影响。 方法 低氧处理体外培养的大鼠肺动脉平滑肌细胞和肺微血管内皮细胞12 h、24 h和48 h,采用原位杂交和免疫组化法检测细胞中FKN mRNA和蛋白的表达,酶联免疫法检测细胞上清液中FKN的浓度。 结果 ①与对照组比较,大鼠肺动脉平滑肌细胞在低氧处理12 h后,其FKN mRNA和蛋白表达、细胞上清液中FKN浓度无明显变化（P>0.05）,低氧24 h后FKN mRNA和蛋白表达、细胞上清液中FKN浓度均增加（P<0.05）,48 h后其增加则更加明显（P<0.01）。②大鼠肺微血管内皮细胞在低氧处理12 h、24 h和48 h后,其FKN mRNA和蛋白表达及细胞上清液中FKN浓度与对照组比较均无明显变化（P>0.05）。 结论 低氧刺激增加了大鼠肺动脉平滑肌细胞表达和分泌FKN。     

乙酰肝素酶在卵巢癌中的表达及其与肿瘤血管生成的关系

目的探讨乙酰肝素酶mRNA及蛋白在卵巢癌组织中的表达及其与肿瘤血管形成的关系。方法采用原位杂交及免疫组化S-P法检测41例卵巢癌组织中乙酰肝素酶mRNA及蛋白的表达,并用免疫组化法对全部标本用CD34抗体进行肿瘤血管内皮染色,计数肿瘤微血管密度（MVD）,结合卵巢癌组织分化程度、组织学类型、临床分期及年龄,探讨乙酰肝素酶与肿瘤血管形成的关系。结果（1）乙酰肝素酶在卵巢癌组织中表达增高．且其mRNA及蛋白的表达具有一致性。（2）临床晚期标本（Ⅲ～Ⅳ期）中的乙酰肝素酶蛋白表达及MVD值高于临床早期（Ⅰ～Ⅱ期）（P〈0．05）;（3）高一中分化癌组与低分化癌组相比较,乙酰肝素酶蛋白表达及MVD值差异有统计学意义（P％0．05）;（4）而组织学类型、年龄对乙酰肝素酶蛋白表达及MVD值无影响（P〉0．05）。（5）临床晚期（Ⅲ～Ⅳ期）组中乙酰肝素酶蛋白的表达与MVD值成正相关（r=0．356,P〈0．05）。结论乙酰肝素酶在卵巢癌中表达增高,且与临床病理分期密切相关,乙酰肝素酶具有促进卵巢癌血管生成的作用。     

氧自由基对豚鼠耳蜗微血管内皮细胞凋亡的影响及其机制

目的 研究氧自由基诱导豚鼠耳蜗微血管内皮细胞凋亡及其可能机制.方法 以黄嘌呤氧化酶作用于黄嘌呤而产生氧自由基.不同浓度氧自由基作用于豚鼠耳蜗微血管内皮细胞,用流式细胞术检测凋亡细胞的百分比;用免疫细胞化学法检测Bax及Bcl-2蛋白表达;用RT-PCR方法检测Bax及Bcl-2 mRNA的表达.结果 氧自由基作用后,与空白对照组相比,各个浓度的氧自由基作用组豚鼠耳蜗微血管内皮细胞凋亡率均明显升高(均P<0.01),同时,BaxmRNA及蛋白的表达均显著增强(P<0.05或P<0.01),Bcl-2 mRNA及蛋白的表达明显减弱(均P<0.01).结论 氧自由基可诱导豚鼠耳蜗微血管内皮细胞发生凋亡,其发生机制与其调节Bax/Bcl-2 mRNA及蛋白的表达有关.     

乙酰肝素酶在卵巢癌组织中的表达及其临床意义

目的 探讨乙酰肝素酶在卵巢癌组织中的表达及其与患者年龄、临床病理特征及预后的关系.方法 采用免疫组化S-P法检测41例卵巢癌组织及10例正常卵巢组织中乙酰肝素酶蛋白的表达,并结合卵巢癌的组织类型、分化程度、临床分期及预后进行分析.结果 ①乙酰肝素酶蛋白在卵巢癌组织中的阳性表达率为80.48%,在正常卵巢组织中为10.00%(P＜0.05).②临床Ⅲ～Ⅳ期卵巢癌中乙酰肝素酶蛋白的表达高于Ⅰ～Ⅱ期(P＜0.01);低分化程度的卵巢癌中乙酰肝素酶蛋白的表达高于高-中分化者(P＜0.05).③各组织类型卵巢癌中乙酰肝素酶蛋白的表达无显著性差异(P＞0.05).结论 乙酰肝素酶在卵巢癌发展过程中起重要作用,检测乙酰肝素酶蛋白的表达有望成为判断卵巢癌预后的标志.     

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.