环缩酚肽抑制小鼠血管增殖性视网膜病变的研究

【目的】观察两种环缩酚肽的衍生物(Hep-A)和(Hep-B)对氧诱导的血管增殖性视网膜病变的抑制作用。【方法】将鼠龄为7d(P7)的C57BL/6幼鼠置于体积分数75%的氧气箱中连续生活5d,建立氧诱导的血管增殖性视网膜病变模型。在12d(P12)幼鼠回到正常大气环境中,同时开始给小鼠皮下注安慰剂(第1组,n=23)、Hep-A10mg/kg(第2组,n=22)、或者Hep-B10mg/kg(第3组,n=22),每天两次,持续5d。在17d(P17)取鼠眼进行冰冻切片和GSA(griffoniasimplicifolialectinB4)染色,或作荧光素灌注和视网膜平铺片,用图象分析软件定量计算视网膜无灌注区和新生血管的面积。【结果】病理检测视网膜新生血管面积:第一组为(0.51±0.08)mm2/鼠(n=7);第2组为(0.11±0.01)mm2/鼠(n=8);第3组为(0.16±0.02)mm2/鼠(n=8)。视网膜平片检测视网膜新生血管面积:第1组为(1.36土0.02)mm2/眼(n=32),第2组为(0.19±0.01)mm2/眼(n=28),第3组为(0.07±0.01)mm2/眼(n=28);视网膜平片检测视网膜无灌注区面积:第1组为(2.33±0.08)mm2/眼,第2组为(1.08±0.09)mm2/眼,第3组为(1.22±0.11)mm2/眼。经ANOVA分析,第2组和第3组分别与第1组比较,视网膜新生血管的面积和视网膜无灌注区的面积,均明显减少,差异有统计学意义(P<0.01)。【结论】两种环缩酚肽衍生物均能强效     

早产儿视网膜病变动物模型的简易制备

目的:采用经济,简易的方法制备早产儿视网膜病变(Retinopathy of prematurity,ROP)的可量化动物模型.方法:新生鼠仔44只称重随机分为2组,实验组于出生后7d置于75%±1%氧箱内饲养5d然后回到正常空气饲养5d,对照组于正常环境饲养.ADP酶法视网膜铺片了解视网膜血管的改变,组织切片HE染色计数突破视网膜内界膜的血管内皮细胞核数,免疫组化法观察视网膜各层血管内皮生长因子(Vascular endothelial growth factor,VEGF)的表达.结果:持续高浓度氧使鼠仔视网膜血管收缩变窄甚至闭塞,视网膜中央部大片无灌溉区,相对缺氧使视网膜大血管明显扩张、迂曲,大量结构异常的新生血管形成.实验组平均每个切面突破视网膜内界膜的内皮细胞核数为(32.800 0±0.767 8)个,对照组平均为(2.750 0±0.716 4)个,两者比较差异有显著性意义(P<0.01);对照组VEGF染色较实验组减弱.结论:该模型可重复性强、稳定性高、可定量研究.是研究早产儿视网膜病新生血管发生机制及相关治疗的合适模型.     

地塞米松抑制视网膜新生血管及血管内皮生长因子表达的实验研究

目的:研究地塞米松(Dexamethasone)在小鼠血管增生性视网膜病变中不同时期的给药效果,及其与血管内皮生长因子(VEGF)的相互作用。方法:将20只7 d龄C57BL/6J幼鼠随机分为4组:正常对照组、模型组、早治疗组和晚治疗组。除对照组外,均建立高浓度氧气诱导的C57BL/6J幼鼠增殖性视网膜病变模型。后2组分别于出生后第7d和第12d起连续5d皮下注射地塞米松注射液(0.5mg/kg.d-1)。17d龄时取幼鼠双眼球作普通病理切片及免疫组化检测,分别检测视网膜新生血管芽内皮细胞核数目及VEGF的表达。结果:模型组视网膜新生血管芽内皮细胞核数目及VEGF的表达较正常对照组明显增加(P<0.01);早治疗组与晚治疗组的视网膜新生血管芽内皮细胞核数目和VEGF表达,较模型组均明显减少(P<0.01);早治疗组较晚治疗组也明显减少(P<0.01)。结论:在氧诱导的血管增生性视网膜病变模型中,地塞米松可以抑制视网膜新血管形成和VEGF的表达,而且病变早期应用地塞米松治疗效果明显优于晚期。     

定量氧致SD大鼠视网膜新生血管模型的制备

目的建立可进行定量研究的视网膜新生血管大鼠模型,为今后视网膜新生血管相关疾病的机制及药物干预研究提供稳定的模型。方法80%高氧环境下饲养新生7 d的SD大鼠,持续5 d后返回正常空气环境中,另一组同龄幼鼠置于正常空气环境中饲养作为对照,鼠龄17 d时处死幼鼠,取眼球做组织切片雪夫高碘酸-苏木素染色,观察并计数突破视网膜内界膜且与内界膜有联系的视网膜新生血管内皮细胞核数。结果与正常对照组相比,高氧诱导组大鼠玻璃体腔内出现视网膜新生血管且管腔扩张。组织切片见实破内界膜且与内界膜相连的视网膜新生血管内皮细胞核数,高氧诱导组平均每张切片为(98.68±10.13)个,两组相比差别有显著统计学意义(P<0.01)。结论该大鼠模型中视网膜新生血管的发生与相对缺氧有关,模型具有可重复性强,可定量研究的优点,是进行视网膜新生血管发生机制及药物干预研究的合适模型。     

尼莫地平与生长因子对视网膜新生血管形成的相互作用

目的:研究钙通道拮抗剂尼莫地平与血管内皮生长因子(VEGF)及血小板洐生生长因子(PDGF)在视网膜新生血管形成中的相互作用.方法:将SD幼鼠随机分为5组:正常对照组,模型组,球后注射药物大剂量组、中剂量组及小剂量组.建立高浓度氧气诱导的SD幼鼠增殖性视网膜病变模型(OIR),后3组分别双眼球后注射尼莫地平(0.2mg/ml)各10、5、2μl,隔日1次×3次.取实验幼鼠眼球作普通病理切片及免疫组织化学检测,分别检测视网膜新生血管芽内皮细胞数目及VEGF与PDGF的表达.结果:模型组视网膜新生血管芽内皮细胞数目及VEGF和PDGF的表达较正常对照组明显增加(P<0.01),双眼球后注药大、中剂量组较模型组均明显减少(P<0.01),小剂量组则无明显变化.结论:VEGF和PDGF能够促进视网膜新生血管形成,一定剂量的钙通道拮抗剂尼莫地平则能抑制视网膜新生血管的形成.尼莫地平在某种程度上能抑制VEGF及PDGF的表达.     

尼莫地平和血小板衍生生长因子在增殖性视网膜病变中的作用

目的:研究钙通道拮抗剂尼莫地平对增殖性视网膜病变(OIR)的治疗作用及其与血小板衍生生长因子(PDGF)的相互作用.方法:以高浓度氧气诱导SD幼鼠OIR模型.将P2(出生后第2天) SD幼鼠随机分为8组(n=10),A为正常对照组,于自然空气中饲养,B为单纯OIR组,C、D、E为球后注射药物组,F、G、H为腹腔内注射药物组,后7组动物均于高浓度氧环境下饲养.球后及腹腔内注射组分别注射不同剂量的尼莫地平.取幼鼠眼球作普通病理切片及免疫组织化学检测,分别检测视网膜新生血管芽内皮细胞数目及PDGF的表达.结果:单纯OIR组视网膜新生血管芽内皮细胞数目及PDGF的表达较正常对照组明显增加(P<0.01),球后注药大、中剂量组较单纯OIR组均明显减少(P<0.01),小剂量组则无明显变化.腹腔内注射各剂量组较未用药组均明显减少(P<0.01).结论:PDGF在OIR组织中表达明显增加,而钙通道拮抗剂尼莫地平可抑制OIR的发生,并在某种程度上抑制PDGF的表达.     

临床用氧对新生鼠视网膜发育的影响

目的 研究不同临床用氧方式对新生鼠视网膜血管发育的影响以及相应视网膜血管内皮生长因子(VEGF) 的表达以探讨ROP发病机制,为早产儿临床合理用氧提供实验依据.方法 新生7 d Wistar鼠仔150只,按不同吸氧方式称质量随机分为5组.实验组1: FiO2由75%分5 d逐渐降低直至停氧(每天下降幅度约为10%);实验组2:吸入氧浓度( FiO2 )5 d逐渐由30%升高至75%后突然中断吸氧.实验组3:持续吸75%高氧5 d后骤然停氧;实验组4:持续吸75%高氧5 d后分5 d逐渐降低FiO2 直至停氧;对照组:置于同实验条件下的空气中饲养.视网膜铺片(ADP酶染色法)了解视网膜血管形态改变;眼球切片(HE法)观察视网膜血管的增生和发育情况;免疫组化法观察5组标本视网膜各层VEGF的表达,并进行视网膜组织内VEGF的含量,血管内皮细胞核两者之间相关性分析.结果 ①视网膜铺片和眼球切片HE染色显示实验组2、3视网膜存在缺氧并出现异常新生血管生成的类似ROP表现.②免疫组化结果显示缺氧后视网膜组织内VEGF的表达明显增加,主要分布在视网膜节细胞层和内核层的毛细血管内皮细胞细胞质和细胞核中.视网膜新生血管芽细胞核数(以与内界膜相关联的突破内界膜的内皮细胞核数C表示)与视网膜组织VEGF含量呈正相关(r=0.807,P＜0.01).结论 吸入高浓度氧后突然停氧可严重影响新生鼠视网膜血管的发育,产生类似早产儿视网膜病的病变.因此临床上应严密监测用氧情况,采取逐渐降低氧浓度的吸氧方式.     

视网膜白蛋白检测在评价小鼠血-视网膜屏障中的实验研究

目的：建立氧诱导视网膜新生血管小鼠模型,观察白蛋白在正常鼠和模型鼠视网膜中的含量变化,探讨白蛋白检测在评价血-视网膜屏障中的作用。方法取鼠龄7 d（P7）的健康C57BL/6J小鼠按随机数字表法分为模型组和正常对照组,模型组置于75％±2％氧气浓度的密闭氧舱中5 d,鼠龄12 d时（P12）返回正常空气环境以建立氧诱导鼠视网膜新生血管模型;正常对照组置于正常空气环境中饲养不予任何处理。模型组和正常对照组均在鼠龄17 d（P17）行FITC-Dextran左心室灌注视网膜铺片,荧光显微镜下观察视网膜血管分布和形态;组织切片HE 染色观察突破视网膜内界膜的血管内皮细胞。提取不同鼠龄（ P13、P15、P17）模型组和正常对照组视网膜总蛋白质,采用western blot技术检测视网膜组织中白蛋白的表达。结果荧光造影结果显示模型组视网膜有大量新生血管形成及明显的荧光素渗漏;组织切片显示模型组有大量突破视网膜内界膜的新生血管内皮细胞核;模型组视网膜白蛋白含量随缺氧时间延长而增加。结论在氧诱导鼠视网膜新生血管形成过程中白蛋白含量明显增加,其变化趋势与血-视网膜屏障破坏和视网膜新生血管形成具有时间上的一致性,白蛋白检测可以作为评价血-视网膜屏障有效指标。     

色素上皮衍生因子对鼠视网膜新生血管形成的保护作用

目的本课题通过小鼠视网膜新生血管动物模型观察玻璃体内注入色素上皮衍生因子(PEDF)对视网膜新生血管形成的影响,并探讨其治疗作用。方法选取鼠龄为7天的健康昆明小鼠28只(56只眼),随机分为4组:正常组、高氧组、PEDF干预组和对照生理盐水组,每组7只。正常对照组鼠在正常空气环境中饲养,其余3组鼠置于氧箱中,建立高氧诱导的视网膜新生血管动物模型。正常组鼠不做任何处理,PEDF干预组幼鼠在出生后第12天向玻璃体腔内注射PEDF 1μl,而对照生理盐水组注射相同体积的生理盐水。各组小鼠均在出生后第17天处死,摘除眼球制作标本,进行组织病理学观察。在光学显微镜下观察并计数突破视网膜内界膜的血管内皮细胞细胞核数目。采用CD31分子免疫组化方法鉴别视网膜内界膜与血管内皮细胞。结果正常对照组标本HE染色未见突破内界膜的内皮细胞核,高氧组和对照生理盐水组标本可见较多的突破内界膜的内皮细胞核,与正常组比较差异具有显著性(P<0.05),PEDF干预组的数目明显少于高氧组和对照生理盐水组,差异均具有显著性(P<0.05),且未见明显药物毒性反应。CD31分子免疫组化染色证实突破内界膜的细胞为血管内皮细胞。结论玻璃体内注入一定剂量的PEDF,能够有效抑制高氧诱导下的小鼠视网膜新生血管形成,PEDF有望成为防治血管增生性视网膜病变的一种有效的方法。     

围产期监护和新生儿

041 8 61　高氧诱导新生鼠视网膜病变的实验研究 /石文静…∥中华围产医学杂志 — 2 0 0 4 ,7( 3) —1 68～ 1 70目的 :建立早产儿视网膜病 (ROP)的动物模型 ,为研究发病机制及治疗提供实验基础。方法 :选择 7日龄新生小鼠 5 4只 ,分为实验组和对照组。实验组 2 7只 ,在氧浓度为 75 %的密闭容器中生长5d ,然后回到正常空气中 ;对照组 2 7只 ,吸室内空气。观察两组视网膜病变 ,视网膜铺片经ADP酶染色 ,以了解视网膜血管的改变 ;视网膜切片经常规HE染色 ,计数突破视网膜内界膜的血管内皮细胞核数目 ,以定量反映视网膜血管的增生情况。结…     

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.