实验性大鼠脊髓损伤神经细胞凋亡的研究

目的 探讨脊髓损伤（SCI）后神经细胞凋亡时间和空间的分布规律。方法 制作大鼠程控电磁吸铁棒下落打击脊髓损伤模型,且末端脱氧核苷酸转移酶介导生物素标记（TUNEL）技术检测细胞凋亡。结果 脊髓损伤后1～2d灰质区TUNEL阳性细胞数最多,随着时间的延长,TUNEL阳性细胞逐渐减少。白质区伤后1d TUNEL阳性细胞数量多,至伤后2d明显减少,伤后7d白质区又出现高峰,至30d白质和灰质区仅见少量散在的TUNEL阳性细胞。结论 脊髓损伤后存在神经细胞的凋亡,白质区和灰质区细胞凋亡演变过程不同。     

鼠缺血脑组织中Fas抗原表达及细胞凋亡的动态研究

目的：探讨脑缺血后Fas抗原表达及与细胞凋亡的关系。方法：以鼠局灶脑缺血为动物模型，在缺血后不同时间点(30min,60min,24h,3d,7d）分别以免疫组化法和TUNEL法动态检测Fas抗原表达和细胞凋亡情况。结果：缺血后30min,Fas抗原呈阳性表达，60min至高峰，24h开始回落，3d时忆为阴性。TUNEL阳性细胞于缺血后60min开始出现，3d达高峰，1周时有回降。结论：脑缺血后有细胞凋亡发生，Fas抗原的大量表达与神经细胞凋亡有密切关系。     

Fas蛋白在鼠急性脊髓组织损伤中分布特点的初步研究

目的 :探讨大白鼠急性脊髓损伤后损伤区Fas蛋白的表达及其分布特点。方法 :Wistar雌性大鼠 4 2只 ,使用改良Allen氏法制作急性脊髓损伤模型 36只 ,采取HE、Nissl染色和免疫组化方法 ,检测损伤后 1 ,4 ,8,2 4 ,72h及 7d的脊髓组织。结果 :在损伤后 4h ,损伤段灰质可见Fas阳性细胞 ,8h表达达高峰。正常组未见Fas阳性细胞表达。高倍镜观察 ,阳性细胞具有凋亡细胞的特征。结论 :Fas蛋白可能在诱导脊髓损伤后神经细胞凋亡的过程中 ,发挥着重要作用     

大鼠坐骨神经损伤修复术后相应脊髓节段Bcl-2、Bax蛋白表达及神经细胞凋亡

目的探讨成年大鼠坐骨神经损伤修复术后相应脊髓节段Bcl-2、Bax蛋白表达及神经细胞凋亡为临床治疗周围神经损伤提供实验依据。方法选取健康成年大鼠68只,随机分为正常对照组、假手术组和实验组,分别于术后不同时间取相应脊髓节段做HE染色、TUNEL染色、免疫组化染色,进行脊髓切片形态观察、检测神经细胞的凋亡及脊髓Bcl-2、Bax蛋白表达。结果假手术组术后无阳性变化。实验组术后脊髓前角运动神经元胞体数目减少,1d后可见脊髓内TUNEL阳性细胞,14d后表达至高峰。1d后,凋亡相关蛋白Bcl-2、Bax表达,14d后达高峰,28d后仍有表达。Bcl-2/Bax值越小,凋亡细胞数量越多。结论大鼠坐骨神经损伤修复术后相应脊髓节段存在神经细胞凋亡,Bcl-2/Bax比值的变化是神经凋亡的重要指标。     

大鼠急性脊髓损伤后神经细胞Bcl-2与Bax比值及细胞凋亡的关系

目的:观察大鼠急性脊髓损伤(ASCI)后不同时间点脊髓神经细胞中Bcl-2和Bax蛋白的表达及神经细胞凋亡的情况. 方法:选取28只SD大鼠,分为对照组(4只)和损伤组(24只),损伤组再分为六个小组,每组4只. 对照组行开椎板术不压迫脊髓,损伤组开椎板压迫脊髓造成大鼠急性损伤模型,损伤组于伤后 4 h, 8 h, 1 d, 3 d, 7 d和14 d 6个时间点取材,做切片备用;免疫组织化学检测Bcl-2, Bax, TUNEL检测细胞凋亡,图像分析各时间段检测结果进行半定量分析;统计学分析各待检数据. 结果:大鼠ASCI后4 h即可见脊髓内TUNEL阳性细胞,8 h表达达高峰,此后表达量逐渐下降,14 d时仍可见少量阳性细胞;大鼠ASCI后凋亡相关蛋白Bcl-2和Bax出现表达,1 d时达高峰,14 d时仍有表达. Bcl-2/Bax值4和8 h时较低以后逐渐增大到一定值,Bcl-2/Bax值越小凋亡的细胞越多. 结论:大鼠ASCI后脊髓内存在神经细胞凋亡,bcl-2和bax相关基因的表达与细胞凋亡数有相关性.     

bFGF预处理对Iso致大鼠心肌损伤及Fas/FasL系统的影响

目的：研究碱性成纤维细胞生长因子(bFGF)对异丙基肾上腺素(Isoprenaline，Iso)致大鼠心肌损伤心肌细胞凋亡及Fas／FasL系统的影响。方法：将Wistar雄性大白鼠随机分成三组：对照组、Iso损伤纽、bFGF预处理组，光镜观察心肌组织的病理变化，TUNEL法检测心肌细胞凋亡，免疫组化和原位杂交方法检测Fas、FasL、Caspase-3蛋白及mRNA的表达水平。结果：Iso损伤组心肌坏死较重，有显著心肌细胞凋亡，且Fas、FasL、Caspase-3基因mRNA及蛋白表达水平明显上调，bFGF预处理组心肌坏死明显减轻，凋亡细胞数减少，Fas、FasL、Caspase-3基因mRNA及蛋白表达水平明显下调。结论：bFGF可通过抑制Fas、FasL、Caspase-3的蛋白及mRNA表达，减少心肌细胞凋亡而减轻保护大鼠的心肌损伤。     

实验性大鼠脑出血细胞凋亡与Bcl-2和Bax表达关系的研究

目的 研究大鼠脑出血（ICH）后血肿周围细胞凋亡及其与凋亡相关蛋白Bcl-2和Bax表达的变化规律和关系.方法 90只SD大鼠随机分为正常对照组（n=10）,假手术组（n=40）,ICH组（n=40）.假手术组和ICH组又依术后12h、1d、2d、3d和7d各分为5个亚组.用胶原酶配合肝素诱导大鼠脑尾状核出血,末端脱氧核苷酸转移酶（TdT）介导的dUTP缺口末端标记法（TdT-mediated-dUTP nick end labeling,TUNEL）观察不同时间点细胞凋亡的变化, SABC法观察Bcl-2和Bax蛋白在各时点表达的变化,免疫印迹（Western-blot）技术检测Bcl-2和Bax蛋白相对光密度比值,电镜观察细胞超微结构改变.结果 ①ICH组TUNEL阳性细胞12h可检测到,3d达高峰,7d仍有表达（P＜0.01）,阳性细胞主要见于血肿周围,多见神经元,有少量神经胶质细胞和血管内皮细胞;②ICH组Bcl-2和Bax蛋白免疫反应阳性细胞分别于出血2、3d达高峰,7d仍有表达,主要表达于血肿区及血肿周围、同侧及对侧皮层;多见神经元,少数为神经胶质细胞、吞噬细胞和中性粒细胞;③Western-blot 正常对照组和假手术组均可检测到Bcl-2、Bax蛋白,但相对光密度比值较小;ICH组Bcl-2相对光密度比值2d达最大值,Bax则3d达最大值,在7d仍明显高于正常对照组（P＜0.05）.结论 ICH后存在细胞凋亡;Bcl-2和Bax参与了ICH细胞凋亡;ICH组TUNEL阳性细胞7d仍可检测到,提示ICH后7d内进行抗凋亡治疗可以不同程度地阻止细胞凋亡的发生.     

大网膜移植抑制大鼠脊髓损伤后神经细胞凋亡的实验研究

目的 探讨脊髓损伤（SCI)后神经细胞凋亡的发生机制和伤后立即行大网膜脊髓移植是否对神经细胞凋亡具有抑制作用。方法 制作程控电磁吸铁棒下落打击脊髓损伤模型,运用末端脱氧核苷酸转移酶介导生物素标记（TUNEL）方法检测细胞凋亡,分别观察SCI后1d、2d、7d组与伤后即刻施行大网膜移植组（存活7d）的细胞凋亡。结果 脊髓灰质区在伤后1d、、2d时TUNEL阳性细胞数最多;脊髓白质区在伤后1d时TUNEL阳性细胞数最多,伤后7d出现第2次高峰;伤后大网膜脊髓移植的TUNEL阳性细胞数较7d明显降低。结论 SCI后白质区和灰质区存在不同演变过程的神经细胞凋亡,SCI后即刻大网膜脊髓移植对细胞凋亡有抑制作用。     

损伤散对急性脊髓损伤大鼠肢体运动功能及Fas,FasL表达的影响

目的 观察损伤散对急性脊髓损伤大鼠运动功能和受损脊髓组织中凋亡相关因子(Fas)及其配体(FasL)表达的影响,并探讨其作用机制.方法 使用改良Allen's法制作大鼠急性脊髓损伤模型,将96只大鼠随机分为空白组、模型组和损伤散组3组,致大鼠脊髓T12—L1节段中度撞击伤,伤后分别于2,8,24 h及7d,比较各组脊髓损伤大鼠的Tarlov评分和损伤脊髓组织中Fas,FasL免疫阳性细胞表达.结果 造模后24 h及7d,损伤散组Tarlov评分显著高于模型组,差异有统计意义(P＜0.05或P＜0.01).与模型组比较,损伤散组造模后8,24 h及7 d Fas,FasL免疫阳性细胞表达差异有统计意义(P＜0.05或P＜0.01).结论 损伤散能促进急性脊髓损伤大鼠运动功能的恢复,使受损脊髓组织中Fas及FasL免疫阳性物质表达降低,对受损脊髓组织有保护和修复的作用.     

心肌细胞凋亡与急性心脏移植排斥的关系

目的：探讨心肌细胞凋亡,Fas蛋白,FasL蛋白表达与心脏移植急性排斥的关系。方法：利用大鼠腹腔异位心脏移植模型。实验组为异基因心脏移植,供体为Wistar大鼠,受体为SD大鼠。对照组为同基因心脏移植,供体与受体均为SD大鼠,设立4个观察点,分别为术后第1,3,5,9天,每个观察点7只动物,应用免疫组化法检测心肌组织Fas蛋白,FasL蛋白的表达,应用3－原位末端标记法（TUNEL法）检测凋亡心肌细胞。结果：（1）实验组移植心肌术后第3天即有一定量TUNEL阳性心肌细胞,术后第5天TUNEL阳性心肌细胞数达高峰,持续至术后第9天,对照组心肌检测到极少量的TUNEL阳性细胞。（2）术后第5,9天标本,实验组检测到大量FasL阳性心肌浸润细胞。（3）实验组与对照组心肌都检测到大量Fas阳性信号。结论：（1）心肌细胞凋亡是心脏移植排斥心肌细胞死亡的一种方式。（2）心脏移植排斥时同时检测到了Fas蛋白,FasL蛋白,TUNEL（＋）心肌细胞,可能是表达FasL的心肌浸润细胞诱导Fas阳性心肌细胞凋亡,介导移植排斥。     

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.