急性高眼压条件下大鼠视网膜节细胞热休克蛋白70的表达

目的 :通过对急性高眼压条件下大鼠视网膜节细胞(RGCs)热休克蛋白 70 (HSP70 )表达变化的分析 ,初步探讨热休克蛋白在高眼压导致视网膜节细胞缺血时的保护作用 .方法 :Wistar大鼠 2 1只一侧眼球在维持 7.98kPa(1mmHg=0 .133kPa)前房压力持续灌注 2h后 ,然后降低眼内压恢复视网膜血液再灌注 ,根据再灌注时间随机分为 0 ,2 ,4 ,8,2 4和4 8h组 ,对照组行前房穿刺 .采用HE染色观察节细胞密度变化情况 ,采用免疫组化方法和图像分析研究视网膜节细胞热休克 70表达及其变化情况 .结果 :再灌注 4 8h组节细胞密度 (3.2 2± 1.6 4个 / 10 0 μm)较其他各组均低 (P <0 .0 5 ) ,对照组和再灌注不同时间后大鼠视网膜节细胞层HSP70表达灰度值之间差异有显著性 ,2 4h组灰度值 (14 8.35 8± 8.0 5 0 )较其他各组低 (P <0 .0 5 ) .结论 :HSP70的表达与缺血再灌注时间关系密切 ,HSP70在视网膜节细胞损伤与防护环节中可能起到重要作用     

水通道蛋白-4在急性高眼压大鼠眼组织的表达变化

目的观察急性离眼压时水通道蛋白-4（AQP4）在大鼠眼组织的表达变化,以探讨AQP4与某些眼压畀常性疾病的关系。方法前房加压灌注法制作大鼠急性高眼压模型;应用免疫组化和原位杂交法观察AQP4庭其mRNA在眼组织的表达变化,并进行半定量分析。结果与对照组相比,AQP4及其mRNA在急性高眼压大鼠视网膜上的表达明显增强,主要集中在节细胞层、内核层。结论急性高眼压状态下,AQP4反其mRNA在视网膜上的表达增加,提示AQP4可能参与了高眼压所致视网膜损伤的病理生理过程。     

肝细胞生长因子在原发性开角型青光眼视神经损伤中的作用及机制研究

目的 探讨肝细胞生长因子( HGF)的受体间质表皮转化因子(c-met)激活是否能促进视网膜神经节细胞存活和再生,以及HGF在慢性青光眼大鼠模型中可能的神经保护作用机制.方法 SD大鼠随机分成3组:正常对照组、慢性高眼压组、慢性高眼压HGF注射组;灼烧3条巩膜上静脉制备慢性青光眼模型;烧灼巩膜上静脉术后立即在玻璃腔内注射4 μl HGF(1 μg/μl);iCare眼压计检查大鼠的眼内压变化;ELISA法测定前房内HGF水平;免疫染色检查c-met在视网膜细胞中的分布;使用末端脱氧核苷酸转移酶dUTP缺口末端标记( TUNEL)染色观察视网膜神经节细胞的凋亡;Western blot法检测蛋白激酶B(Akt)、磷酸化蛋白激酶B( p-Akt)、B细胞淋巴瘤基因-2(Bcl-2)和抗凋亡蛋白B细胞淋巴瘤/白血病-xl(Bcl-xl)蛋白的表达水平.结果 术后早期眼压显著升高,并维持2周;与正常组相比,慢性高眼压大鼠组前房HGF明显升高;与慢性高眼压组相比,慢性高眼压注射HGF组TUNEL标记的RGC减少,差异有统计学意义( P＜0. 05);在视网膜中,c-met主要在神经节细胞(RGC)层的细胞中表达;在视网膜样本中,与正常对照组比较,慢性高眼压组p-Akt、Bcl-2和Bcl-xl蛋白表达显著下调(P＜0. 05);与慢 性高眼压组相比,慢性高眼压 HGF 注射组 p-Akt、Bcl-2 和Bcl-xl蛋白表达显著上调(P＜0. 05).结论 HGF可能通过激活Akt信号通路以及增加其下游的相关抗凋亡蛋白表达而发挥对慢性高眼压视神经的保护作用.     

利鲁唑对急性高眼压致大鼠视网膜损伤的保护作用

目的 探讨腹腔内注射利鲁唑对急性高眼压所致的大鼠视网膜损伤的保护作用.方法 将120只大鼠分为正常对照组、高眼压对照组及利鲁唑干预组,每组40只.采用前房灌注的方式建立急性高眼压模型,免疫组化及末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记法(TUNEL) 分别测定3组大鼠再灌注后12、24、48、72 h的Caspase-3蛋白表达情况及大鼠视网膜各层细胞的凋亡水平,HE染色观察各组大鼠视网膜结构改变.结果 与高眼压对照组相比,利鲁唑干预组视网膜增厚及结构紊乱情况明显减轻,不同时间点Caspase-3的表达水平及凋亡细胞数明显降低(P< 0.05).结论 利鲁唑可减少大鼠急性高眼压损伤后视网膜神经节细胞凋亡,对视网膜具有一定的保护作用.     

急性高眼压大鼠视网膜睫状神经营养因子表达变化

目的探讨睫状神经营养因子（CNTF）在急性高眼压大鼠视网膜中的定位及表达变化。方法采用前房灌注法建立大鼠急性高眼压模型,在1,3,7,14,21 d分别摘取眼球,运用RT-PCR和免疫组织化学方法检测视网膜CNTF的定位及表达变化。结果对照组大鼠视网膜神经节细胞层（GCL）有少量CNTF表达,急性高眼压大鼠视网膜CNTF的表达显著增加,建立模型后第3-7天表达量达到高峰,此时除GCL外,神经纤维层（NFL）、内核层（INL）亦发现CNTF表达,后表达减少。结论急性高眼压大鼠视网膜CNTF表达增加,可能是大鼠高眼压损伤后的一种保护性反应,CNTF的增加对视网膜神经节细胞可能有保护作用。     

急性高眼压后大鼠视网膜HO-1表达的变化

目的研究急性高眼压后大鼠视网膜血红素氧合酶-1(HO-1)表达的变化规律,为视网膜损伤后的治疗提供理论依据。方法采用前房灌注加压法制作急性高眼压大鼠模型,运用实时定量逆转录聚合酶链反应方法测定HO-1mRNA的表达情况。结果急性高眼压后1d视网膜HO-1mRNA的表达明显升高且达高峰,3d时表达水平下降,7d时再次升高,14d时继续升高。结论急性高眼压造成视网膜氧化应激损伤可以诱导HO-1表达增高。     

青光安颗粒剂对实验性高眼压大鼠视网膜节细胞代谢作用的研究

采用细胞色素氧化酶组织化学法观察青光安颗粒剂对实验性高眼压大鼠视网膜节细胞代谢细胞的影响。结果表明：青光安颗粒剂治疗的高眼压模型眼视网膜ＣＯ活性节细胞和高ＣＯ活性节细胞密度与对照组高眼压模型眼比较，有非常显著性差异。     

实验性局灶性脑缺血再灌注Fos蛋白的表达

目的　探讨大鼠局灶性脑缺血再灌注Fos蛋白表达的意义及其机制。方法　采用免疫组化法观察Fos蛋白在大鼠局灶性脑缺血再灌注后皮层和基底节区的动态变化。结果　Fos蛋白在大脑中动脉阻断 1h后 ,随再灌注时间延长其分布不同。基底节区持续时间长 ,阳性细胞多 ,而皮层持续时间短 ,阳性细胞少。其中再灌注 90min ,Fos蛋白表达最显著。大脑中动脉供血中心区皮层几无表达。结论　Fos蛋白的表达与神经细胞存活密切相关 ,出现Fos蛋白表达的区域神经元易于耐受缺血性损害而存活 ,可能是神经细胞自我保护机制之一 ,Fos蛋白的表达存在扩散抑制     

急性高眼压兔眼视网膜中过氧亚硝基阴离子的表达及葛根素对其表达的影响

目的探讨过氧亚硝基阴离子在急性高眼压兔眼视网膜中的表达及其损伤机制,并评价葛根素对其表达的影响。方法27只新西兰大白兔,随机分为正常对照组、模型组及治疗组(耳缘静脉注射葛根素),通过前房灌注建立急性高眼压模型,观察视网膜形态学变化,采用原位缺口末端标记法检测高眼压后6、12、247、2 h视网膜中凋亡细胞,应用免疫组织化学方法检测视网膜中过氧亚硝基阴离子的表达。结果模型组6 h时神经纤维层和内丛状层明显水肿,24 h视网膜变薄、神经节细胞变性、核固缩;凋亡细胞于6 h时少量表达,24 h达到高峰,72 h减少;过氧亚硝基阴离子的表达趋势与凋亡细胞一致。治疗组视网膜水肿及神经节细胞变性、核固缩程度较轻;细胞凋亡及硝基酪氨酸的表达趋势同模型组,但表达均明显低于模型组(P<0.05)。结论急性高眼压兔眼视网膜中过氧亚硝基阴离子的表达增多,并参与介导神经节细胞凋亡,葛根素通过其抗氧化作用抑制过氧亚硝基阴离子的表达,在一定程度上保护神经节细胞。     

急性高眼压t-PA表达的实验研究

目的:探索血浆组织型纤溶酶原活化因子(t-PA)在青光眼视网膜中的表达变化、可能的作用以及脑源性神经营养因子(BDNF)对其的影响。方法:采用前房灌注法建立兔急性高眼压模型,在玻璃体内注射BDNF或BSS液,用免疫组织化学法检测视网膜t-PA的蛋白表达与定位,用Westernblot分析检测视网膜t-PA的蛋白表达。结果:t-PA与β-actin灰度比值,正常对照组为(57.95±3.79)%、BSS组为(89.36±3.59)%、BDNF组为(66.66±2.16)%;BSS组与正常对照组和BDNF组相比,t-PA表达明显升高(P<0.05)。高眼压后视网膜上t-PA的免疫组化染色明显增强,神经纤维层、RGCs层呈强阳性染色。结论:t-PA与青光眼的病理过程有相关性,BDNF可能部分通过抑制t-PA的表达发挥神经元保护作用。     

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.