一氧化氮在糖尿病大鼠血和肾组织中的动态变化

目的 :探讨一氧化氮 (NO)在糖尿病大鼠血和肾组织中的动态变化。方法 :用链佐脲菌素 (STZ)制造大鼠糖尿病模型 ,成模后 4、8、12周分别采用硝酸还原酶间接法和吸光度比较法测定血清和肾组织NO含量、一氧化氮合酶 (NOS)活性。同时取肾组织进行光镜和透射电镜检查。结果 :糖尿病大鼠血清和肾组织NO含量、NOS活性 4周时升高 ,8、12周时逐渐下降 ,与对照组比较差异有显著统计学意义 (P <0 .0 5 )。肾组织图象定量分析显示 ,糖尿病组肾小球平均截面积和基底膜厚度高于对照组 (P <0 .0 5 )。相关分析表明 ,肾组织中NO与GFR呈正相关 ,与尿蛋白排泄量、基底膜厚度呈负相关。结论 :糖尿病大鼠血和肾组织中NO含量早期升高 ,随病程延长逐渐降低 ,与NOS活性变化一致。内皮源性舒张因子NO与糖尿病早期肾小球高滤过和肾脏形态学改变有关。     

糖尿病大鼠肾组织TGF-β1及细胞外基质表达

目的观察糖尿病早期大鼠肾组织TGF-β1的表达与肾功能改变的相关性.方法用AV1000全自动生化分析仪观察链脲菌素实验性大鼠肾功能变化情况;取糖尿病2、4周大鼠双肾称重后取材,常规制片HE及PAS染色观察组织学改变;免疫组化采用LSAB法分别测实验组及对照组大鼠肾组织TGF-β1、FN 、LN的表达.结果糖尿病组大鼠肾功能障碍,部分指标糖尿病4周组比糖尿病2周组显著增高(P＜0.05).组织学观察糖尿病组大鼠亦显示肾小球肥大,系膜区增宽及部分肾小球囊壁、近曲小管基底膜出现PAS阳性均质蛋白性物质.结论 DM大鼠肾小球TGF-β1表达上调,与两种ECM成分LF、FN过度聚集呈显著正相关,证实了TGF-β1对ECM的异常调节和促进肾小球硬化的作用.     

OLETF糖尿病大鼠心肌光镜与透射电镜病理改变的观察

目的 使用光镜与透射电镜,观察OLETF糖尿病大鼠在糖尿病病程中晚期的心肌病理改变.方法 以7只OLETF糖尿病大鼠作为实验动物模型,做OGTT试验诊断并了解糖尿病病程.另以7只LETO大鼠为对照.鼠龄58周时检测血浆血脂、胰岛素、C-肽水平,评估胰岛功能及脂代谢状况.股动脉插管测量血压、颈动脉逆行插管入左心室测量左室内压以评估心功能.石蜡切片HE、PAS、VG染色观察心肌光镜组织结构.两组各取3只大鼠超薄切片透射电镜观察心肌超微结构,测量心肌壁毛细血管基膜厚度并进行比较.结果 OGTT试验显示34周后80%OLETF糖尿病大鼠已发生糖尿病,鼠龄58周时三酰甘油、胆固醇较对照组明显增加(P＜0.05).左心室压下降速率(-dp/dt)低于对照组(P＜0.05).OLETF糖尿病大鼠HE染色显示心肌细胞肥大,肌纤维收缩呈波浪状;PAS,VG染色显示心肌间质胶原纤维增多.超薄切片透射电镜观察显示心肌细胞挛缩、肌原纤维收缩带形成,心肌间质胶原沉积,毛细血管基膜增厚.OLETF糖尿病大鼠的基膜厚度为(64.33±4.38)nm,较LETO对照组大鼠(46.09±3.62)nm明显增厚(P＜0.001).结论 58周龄OLETF糖尿病大鼠的心肌病理改变为心肌细胞挛缩、心肌间质胶原沉积和毛细血管基膜增厚,并与左心室舒张功能减退同时存在.     

血栓调节蛋白和纤溶酶原激活物抑制物-1在糖尿病肾病中的作用

糖尿病肾病(DN)是糖尿病(DM)常见而严重的并发症之一,其病理特征为肾小球基底膜增厚、系膜内玻璃样物质增生、内皮细胞损伤与激活。DN患者肾脏局部纤溶酶原激活物抑制物-1(PAI-1)表达增多,抑制了肾小球和肾小管间质细胞外基质(ECM)的降解,导致ECM进行性堆积,肾小球基底膜增厚,肾小球硬化。DM并发微血管病变尤其是DN时,血浆血栓调节蛋白(PTM)水平显著升高。内皮损伤伴随DN的整个发展过程,它的出现甚至早于DN微量白蛋白尿的出现。测定PAI-1、TM水平对早期诊断、早期治疗DN有重要临床意义。     

骨形态形成蛋白-7在实验性肾间质纤维化大鼠模型中作用的研究

目的:观察单侧输尿管梗阻(UUO)大鼠模型中骨形态形成蛋白-7(BMP-7)在肾小管间质中的表达部位、动态变化及其与肾间质纤堆化的 关系。方法:60只Wistar大鼠随机分为正常大鼠组、假手术组和单侧输尿管梗阻组,分别于术后1d、3d、7d、14d处死大鼠,采用RT-PCR检测BMP-7mRNA、 转化生长因子-β1(TGF-β1)mRNA表达水平,用免疫组化检测肾小管间质中BMP-7、TGF-β1蛋白的定位和表达及其与输尿管梗阻后肾间质损害的关系。 结果:与时照组相比,BMP-7mRNA于术后第1天即出现表达下降并随梗阻时间延长递减,而TGF-β1mRNA则随着梗阻时间延长增加(P均<0.05)。BMP -7蛋白在对照组中高度表达,髓质区表达明显高于皮质区,主要分布在肾小管及肾间质,肾小球内基本无表达。在梗阻组BMP-7蛋白表达呈进行性下 降,而TGF-β1蛋白表达则逐渐增加(P均<0.01)。结论:BMP-7蛋白于肾小管间质病变早期即出现表达下调,早于肾间质纤维化的出现,且随间质病变进 展进行性下降。BMP-7表达下调可能参与介导肾小管间质损害的发生发展。     

左旋-精氨酸对糖尿病肾病鼠的影响

目的 :了解左旋 精氨酸 (L arginine)对糖尿病 (DM )鼠肾脏的影响并初步探讨其机制。方法 :将实验大鼠分 3组 ,正常对照组 (A组 ) 8只 ;DM组 (B组 ) 8只 ;DM 4周后 +0 .5 %L arginine组 (C组 ) 8只。观察 8周。结果 :第 1周末开始 ,B、C组Ccr和UAE较A组明显升高 ,B、C组之间无显著差异 ;尿NO-2 /NO-3 第 1,2周B、C较A组明显升高 ,B、C组之间无显著差异 ,第 8周B、C组较A组降低 ,但C组较B组升高。第 8周末肾脏病理变化为 ,B组肾小球系膜区增宽 ,肾小球毛细血管基底膜弥漫性增厚 ,系膜及基质PAS阳性物增多 ,但C组上述变化明显较B组轻。ABC染色Laminin阳性物可见B组肾小球系膜区弥漫性增宽 ,肾小管基底膜阳性物呈宽带状沉积 ,而C组肾小球系膜区稍宽 ,肾小管基底膜阳性物呈线状沉积 ,χ2 检验P <0 .0 5。B、C组肾皮质NO-2 /NO-3 与PAS阳性物灰度值呈直线正相关。结论 :糖尿病肾病进程中内源性NO表现为初期的升高到发展中的下降趋势。内源性NO生成增多是导致糖尿病肾病早期肾小球高灌注、高滤过的重要原因之一 ,在糖尿病肾病发展中补充小剂量L arginine能减轻肾小球基底膜的增厚和基质的增生 ,延缓肾小球硬化 ,对肾脏起保护作用。     

立体计量法对链脲佐菌素糖尿病大鼠肾小球毛细血管基底膜和系膜基质改变观察初探

用立体计量法观察糖尿病大鼠肾小球毛细血管基底膜和系膜基质的改变。肾小球毛细血管基底膜和系膜基质随糖尿病病程进展而增加,且系膜基质增多出现在肾小球毛细血管基底膜增厚之前,两者呈正相关,说明本计量法对糖尿病肾病的研究有实用价值。     

厄贝沙坦对STZ诱导的糖尿病大鼠肾脏CTGF、p27kip1表达的影响

目的:观察厄贝沙坦(Irb)对大鼠糖尿病模型肾脏结缔组织生长因子(CTGF)、p27k ip1表达的影响。方法:将SD大鼠分为健康对照组(C组)、糖尿病肾病组(DN组)和糖尿病肾病Irb治疗组(DNI组),DN组和DNI组大鼠制成糖尿病模型,DNI组予以50 mg.kg-1伊贝沙坦灌胃。各组大鼠再分为4个亚组,分别于成模后1、2、4、8周时处死,观察各组大鼠的血糖、体重、24 h尿白蛋白、内生肌酐清除率(Ccr)、肾重、肾脏肥大指数、肾小球面积(AG)和体积(VG)、肾小管面积(AT)、肾小球基底膜(GBM)厚度、肾小管基底膜(TBM)厚度的改变,通过免疫组化观察肾CTGF和p27k ip1的表达。结果:DN组和DNI组大鼠血糖较C组明显升高且维持在一个较高水平(P<0.01)。C组体重增长迅速,DN组和DNI组大鼠体重增长缓慢(P<0.01)。DN组大鼠的24 h尿白蛋白、Ccr、肾重、肾脏肥大指数、AT和VG在糖尿病早期即呈时间依赖性增加(P<0.01或P<0.05)。免疫组化半定量分析显示,各期DN组大鼠肾小球和肾小管的CTGF、p27k ip1表达均高于C组(P<0.01或P<0.05)。8周时DN组大鼠GBM和TBM均较C组明显增厚(P<0.01),而Irb可显著抑制上述参数的增加。CTGF与p27k ip1的表达、24 h尿白蛋白、AT、AG、VG呈显著正相关关系(P<0.05)。结论:早期应用Irb可抑制糖尿病大鼠早期肾脏肥大和CT-GF与p27k ip1的表达,此为早期使用该药预防DN肾脏肥大的发生提供了新的理论依据。     

血糖波动与持续性高血糖对糖尿病大鼠肾脏病理改变及IV型胶原表达的影响

目的: 观察血糖波动与持续性高血糖对糖尿病大鼠肾脏病理改变及 IV型胶原(Col IV)表达的影响。方法: 将60只SD雄性大鼠随机分为正常组和模型组,正常组大鼠喂以普通饲料,模型组大鼠高糖高脂饲料喂养6周后予以小剂量链脲佐菌素(STZ,30mg/kg)皮下注射制造糖尿病大鼠模型。再将糖尿病组大鼠随机分为持续性高血糖组和血糖波动组,其中血糖波动组给予每日两次皮下注射胰岛素人为诱导血糖波动。3个月后将大鼠剖腹取出肾,行HE染色、PAS染色、Col IV免疫组织化学及Western印迹检测。结果: 与正常组大鼠比较,模型组大鼠肾小球体积增大、毛细血管内皮细胞基底膜增厚、系膜基质增多、肾小球球囊腔扩张,肾小管体积增大、肾小管管腔扩张、上皮细胞基底膜增厚,肾小球通透性增强,肾脏病理形态改变明显;肾肥大指数增加、肾小球硬化指数增加、Col IV表达量明显增加(P<0.01)。与持续性高血糖组比较,血糖波动组大鼠上述指标变化更为明显(P<0.05)。结论: 糖尿病大鼠出现了肾小球基底膜增厚、系膜基质增多等病理形态的改变,血糖波动组肾小球硬化更明显, Col IV的增多可能与糖尿病肾病的严重程度有关。     

酸味中药复方对糖尿病大鼠肾小球病变的作用

目的：探讨酸味中药复方对糖尿病肾小球病变的影响。方法：应用免疫组织化学和图像分析法检测了肾小球内高级糖化产物、血小板源生长因子－Ａ以及毛细血管基膜与系膜区的结果变化。结果：糖尿病大鼠肾小球终末糖化产物增加,肾小球血小板源生长因－Ａ明显增高,肾小球发生基膜增厚和系膜增宽的病理变化。结论：酸味中药复方对糖尿病大鼠肾的非酶糖化反应和血管病变有一定的抑制作用。     

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.