脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血细胞凋亡及Caspase-3蛋白表达的影响

目的 探讨人脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血细胞凋亡及Caspase-3蛋白表达的影响.方法 线栓法制作大鼠大脑中动脉缺血(MCAO)2 h再灌注模型.60只健康雄性SD大鼠随机分为4组:正常组(6只),假手术组(6只),缺血对照组(24只)和移植治疗组(24只).后两组又分为缺血2 h再灌7、14、21和28 d组(各6只).体外培养脂肪基质细胞,诱导分化为神经干细胞.造模成功后24h,移植治疗组经尾静脉移植人脂肪组织来源的神经干细胞悬液(细胞浓度为2×106/mL),缺血对照组经尾静脉注射生理盐水,假手术组不做任何处理.TUNEL法检测细胞凋亡,免疫组化SABC法检测Caspase-3表达.结果 与缺血对照组比较,移植治疗组各时间点(7、14、21和28 d)的细胞凋亡数均明显减少(均P<0.01),Caspase-3蛋白表达均明显减少(均P<0.01).结论 脂肪组织来源的神经干细胞可能通过下调Caspase-3蛋白表达,减少局灶性脑缺血细胞凋亡,对脑缺血再灌注损伤后的神经细胞起保护作用.     

大鼠局灶性脑缺血再灌注后VEGF及VEGFmRNA的表达

目的:探讨大鼠局灶性脑缺血再灌注后脑组织内血管内皮细胞生长因子(VEGF)及VEGFmRNA的表达及意义.方法:制备大鼠局灶性脑缺血再灌注模型,动物分为缺血2h再灌注6h、12h、24h、48h、72h及假手术对照组.应用免疫组化法检测VEGF蛋白的表达;RT-PCR法检测VEGF mRNA的动态变化.结果:大鼠缺血再灌注6h,缺血侧脑组织梗死灶周边VEGF免疫阳性反应已出现,24h明显增多,48h达高峰;VEGF mRNA在6h开始上升,12h达高峰,24h开始下降,48～72h接近正常水平.结论:脑缺血再灌注损伤可以诱导VEGF表达增强,提示VEGF对缺血性脑损伤有保护作用.     

高压氧对局灶性脑缺血再灌注大鼠脑神经细胞凋亡及bcl-2蛋白表达的影响

目的:探讨高压氧对局灶性脑缺血再灌注大鼠脑神经细胞凋亡及bcl-2蛋白表达的影响.方法:选取健康雄性Wistar大鼠60只,随机分为假手术组(10只)、缺血对照组(25只)及缺血再灌注加高压氧治疗组(HBO组,25只).采用动脉线栓法制成大脑中动脉(MCA)局灶性脑缺血再灌注模型.用四氮唑染色观察局灶性脑缺血再灌注后120 h各组大鼠脑梗死灶的大小,应用TUNEL法及免疫组化方法检测缺血3 h再灌注6 h、24 h、48 h、72 h、120 h大鼠脑组织神经细胞凋亡及bcl-2蛋白的表达.结果:假手术组未发现脑梗死灶,凋亡细胞及bcl-2蛋白表达阴性.HBO组缺血再灌注后120 h大鼠梗死灶体积(15.9±4.2)明显小于缺血对照组(26.8±4.9)(P＜0.01);HBO组大鼠再灌注24 h,48 h,72 h各时点凋亡指数均低于缺血对照组,bcl-2蛋白表达均高于缺血对照组(P＜0.05).结论:脑缺血再灌注后凋亡细胞增多,bcl-2蛋白表达增强,高压氧治疗可以上调bcl-2蛋白的表达,抑制脑神经细胞凋亡,具有脑神经保护作用.     

血管内皮生长因子在脑缺血耐受大鼠中的表达

目的检测局灶性脑缺血预处理诱导大鼠脑缺血耐受模型再灌注不同时间窗血管内皮生长因子(VEGF)的表达,探讨其在脑缺血耐受中的作用。方法将55只Wistar大鼠随机分成假手术对照组(SS+SS,5只)、假手术+再缺血组(SS+MCAO,25只)、缺血预处理+再缺血组(IP+MCAO,25只),后两组再随机分成5个亚组。线栓法阻塞大脑中动脉(MCAO)建立局灶性缺血预处理模型(缺血预处理10min),分别在缺血预处理后1、3、7、14、21d进行再次缺血2h再灌注22h,然后取脑检查。应用苏木精-伊红染色观察脑组织病理变化并测定脑梗死体积,观察神经行为缺损,免疫组化法检测VEGF蛋白表达。结果IP+MCAO组1、3、7d亚组与SS+MCAO组相应亚组比较,神经行为缺损评分明显降低,梗死体积明显减小,VEGF表达明显增高,差异均有显著性(t=2.357～11.479,P<0.05)。结论缺血预处理诱导了脑缺血耐受,预缺血诱导的VEGF表达增加在脑缺血耐受中发挥重要作用。     

大鼠局灶性脑缺血再灌注脑内p47phox表达变化的研究

目的通过观察大鼠局灶性脑缺血再灌注后p47phox基因在脑内表达动态变化,探讨p47phox在局灶性脑缺血再灌注后对脑缺血损害的作用及机制,可能为p47phox基因作为脑卒中的候选基因提供理论依据。方法采用逆转录聚合酶链反应(RT-PCR)法检测大鼠局灶性脑缺血再灌注后脑缺血再灌注组和假手术组再灌后第1、3、6、12和24h和正常对照组p47phox基因在脑内的表达。结果正常对照组和假手术组大鼠脑皮质内有少量p47phox mRNA表达,组内各时间点比较无统计学意义(P>0.05);脑缺血再灌注组,缺血再灌注1h后p47phox mRNA表达增加,至3h达高峰(与假手术组和正常对照组相比差异显著,P<0.01)。结论p47phox可能参与了脑缺血再灌注后损伤,并在其中发挥了重要作用。     

大鼠脑缺血再灌注损伤后死亡相关蛋白激酶的表达及意义

目的 探讨大鼠局灶性脑缺血再灌注损伤时死亡相关蛋白激酶(Death associated related protein kinase,DAPK)的表达变化及其与脑缺血再灌注损伤的关系。方法 线拴法建立局灶性脑缺血再灌注模型,SD大鼠随机分为正常对照组、假手术组和缺血再灌注组,缺血组于栓塞3h后分别再灌注12h、24h、3d、7d。HE染色观察组织病理变化;免疫组化法检测DAPK的表达;原位杂交法检测DAPK mRNA的表达。结果 缺血组DAPK免疫阳性反应于再灌注后24h开始增强并于3d时达高峰,随后下降;DAPK mRNA的表达于再灌注3d时达高峰,随后下降。结论 脑缺血再灌注损伤诱导DAPK表达增强;DAPK在缺血性脑损伤中起重要作用。     

眼针对局灶性脑缺血再灌注大鼠VEGF/VEGFR系统表达的影响

目的   通过观察眼针对局灶性脑缺血再灌注大鼠血管内皮生长因子及其受体(VEGF/VEGFR)系统表达的影响,探讨眼针促进局灶性脑缺血再灌注大鼠微血管生成的作用机制。方法   采用改良线栓法制备局灶性脑缺血再灌注模型,采用眼针治疗。再灌注3、24、72h后,免疫组化法检测脑微血管密度及VEGF、VEGFR2表达;逆转录-聚合酶链反应法检测大鼠VEGF mRNA、VEGFR2 mRNA表达。结果   与模型组比较,眼针组缺血再灌注24、72h微血管密度明显增加,各时间点VEGF、VEGFR2表达明显增加,VEGF mRNA、VEGFR2 mRNA表达明显上调(P均＜0.05)。结论   眼针可以促进脑缺血再灌注大鼠微血管生成,其机制与上调VEGF/VEGFR表达有关。     

脂肪源性干细胞穴位注射对大鼠脑缺血/再灌注后海马中血管生成的影响

目的探讨脂肪源性干细胞(ADSC)穴位注射对大鼠脑缺血/再灌注损伤后海马中血管生成的影响。方法24只SD大鼠随机分为假手术组、模型组、对照组(穴位注射生理盐水)和治疗组(穴位注射ADSC),每组6只。线栓法制作大鼠大脑中动脉缺血/再灌注模型,治疗组在大鼠大椎、双侧内关穴位注射PKH-26标记ADSC细胞悬液,对照组相同穴位注射同剂量生理盐水。缺血72 h后行神经功能损害评分(NSS),采用免疫组织化学法及实时荧光定量聚合酶链反应检测血管内皮生长因子(VEGF)蛋白和mRNA表达,免疫组织化学法进行CD34染色计数微血管密度(MVD)。结果脑缺血后72 h,治疗组大鼠海马区可见PKH-26标记的ADSC表达,而其他3组未发现红色荧光信号。模型组、对照组和治疗组NSS评分高于假手术组(P<0.05);与模型组比较,对照组和治疗组NSS评分显著降低(P<0.05),其中治疗组NSS评分低于对照组(P<0.05)。假手术组海马区中未见VEGF蛋白和mRNA表达,模型组、对照组和治疗组VEGF蛋白和mRNA表达及MVD较假手术组显著增加(P<0.05),其中治疗组VEGF蛋白和mRNA表达及MVD较模型组和对照组显著增高(P<0.05),而模型组和对照组3个指标比较差异无统计学意义(P>0.05)。结论穴位注射ADSC对大鼠脑缺血/再灌注有神经保护作用,其作用机制可能与促进海马VEGF表达,增加MVD有关。     

实验性大鼠局灶性脑缺血再灌注后VEGF与TGF-β1的表达及意义

目的 研究大鼠局灶性脑缺血后血管内皮生长因子(VEGF)、转化生长因子(TGF-β1)在缺血再灌注不同时程和不同部位的表达及意义。方法 采用线栓法制作大鼠局灶性大脑中动脉闭塞模型，将动物随机分为正常对照组、假手术组、再灌注12h、24h、3d、7d组。采用免疫组化方法观察大鼠缺血再灌注不同时程VEGF、TGF-β1的表达情况。结果 缺血6h再灌注12h VEGF、TGF-β1即有阳性表达，再灌注24hVEGF、TGF-β1表达明显增多，持续至3d，以后表达减少，但7d仍有阳性表达。阳性表达的细胞主要是神经胶质细胞、神经元和血管内皮细胞。各时间段二者的阳性表达主要集中在梗死灶周围区。结论 VEGF、TGF-β1共同参与缺血脑损伤的病理发展及修复过程。     

活血健脑胶囊对大鼠局灶性脑缺血再灌注后神经细胞的保护作用

[目的]研究活血健脑胶囊对大鼠局灶性脑缺血再灌注后神经细胞的保护作用.[方法]选用SD大鼠,随机分为假手术组、模型组、尼莫地平组(剂量为10.8 mg·kg-1·d-1)、活血健脑胶囊组(剂量为1.8 g·kg-1·d-1),后3组再分为缺血2 h及再灌注6、12、24 h组;采用线栓法复制大鼠局灶性脑缺血再灌注模型,免疫组织化学法检测神经细胞细胞色素C(Cyt C),原位杂交法检测Caspase-3 mRNA、Caspase-9 mRNA的表达.[结果]活血健脑胶囊组脑缺血/再灌注6 h细胞色素C阳性弱表达,与模型组比较有显著性差异(P<0.01),脑缺血/再灌注12 h、24 h细胞色素C的阳性表达与模型组及假手术组比较无显著性差异(P>0.05).模型组脑缺血再灌注24 h海马Caspase-3 mRNA、Caspase-9 mRNA的表达达高峰(P<0.01);活血健脑胶囊组脑缺血/再灌注24 hCaspase-3 mRNA、Caspase-9mRNA袁达均减弱,与模型组比较有显著性差异(P<0.01).[结论]活血健脑胶囊对局灶性脑缺血再灌注后神经细胞的保护作用可能与其能影响神经细胞Cyt C的释放,降低脑缺血/再灌注后脑组织Caspase-3、Caspase-9的活性,从而阻断细胞凋亡的发生有关.     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.