DHPG诱导的大鼠海马脑片癫癎样放电模型基因表达谱分析

目的 研究代谢型谷氨酸受体-Ⅰ（mGluR-Ⅰ）激动剂D-对羟基苯甘氨酸（DHPG）诱导的大鼠离体海马脑片癫癎样放电模型的基因表达谱。方法 以含50 μmol DHPG的人工脑脊液持续灌流大鼠离体海马脑片,诱导建立癫癎样放电模型（DHPG组,n=3）。采用cDNA微阵列分析技术获得DHPG组的基因表达谱,与对照组（n=3）比较筛选出差异表达基因,并进行富集功能分类。结果 与对照组比较,DHPG组样本模型分别筛选出的上调基因206个,下调基因489个;其中差异表达达1.5倍的上调基因67个,下调基因86个;2.0倍以上的上调基因6个,0.5倍以下的下调基因25个。富集功能分类结果显示,差异表达基因功能涉及蛋白结合（19个）、分子功能、钙离子结合和核苷酸结合等多方面。结论 癫癎样放电及mGluR-Ⅰ激动剂的作用均涉及多种基因,是一复杂的调控过程。实验为进一步研究提供了依据。     

DHPG和BMI诱导的癫痫样放电模型的建立及比较

目的 在离体大鼠海马脑片上,通过激活Ⅰ型代谢型谷氨酸受体(mGluR)或阻断γ-氨基丁酸(GABA)受体,诱导癫痫样放电.方法 将离体大鼠海马脑片暴露于Ⅰ型mGluR特异性激动剂对羟基苯甘氨酸(DHPG)或GABAA受体阻断剂荷包牡丹碱(BMI),利用全细胞记录模式记录海马CA3区域单个锥体细胞的电活动.结果 暴露于DHPG或BMI中的海马脑片CA3区域的锥体细胞均产生癫痫样放电,且两者的放电频率比较差异无统计学意义(P>0.05).结论 在离体情况下,破坏神经兴奋性和抑制性系统的平衡可以诱导产生癫痫样放电活动.     

DHPG和BMI诱导的癫痫样放电模型的建立及比较

目的在离体大鼠海马脑片上,通过激活Ⅰ型代谢型谷氨酸受体（mGluR）或阻断γ-氨基丁酸（GABA）受体,诱导癫痫样放电。方法将离体大鼠海马脑片暴露于Ⅰ型mGluR特异性激动剂对羟基苯甘氨酸（DHPG）或GABA．受体阻断剂荷包牡丹碱（BMI）,利用全细胞记录模式记录海马CA3区域单个锥体细胞的电活动。结果暴露于DHPG或BMI中的海马脑片CA3区域的锥体细胞均产生癫痫样放电,且两者的放电频率比较差异无统计学意义（P〉0．05）。结论在离体情况下,破坏神经兴奋性和抑制性系统的平衡可以诱导产生癫痫样放电活动。     

大鼠海马神经元癫(癎)模型中神经元凋亡及caspase 3基因的表达

目的:探讨体外培养大鼠海马神经元癫(癎)模型中神经元丢失的分子机制.方法:制备大鼠海马神经元癫(癎)样放电细胞模型,以膜片钳全细胞记录对模型放电进行检测,采用RT-PCR法克隆大鼠全长caspase 3 cDNA,采用原位杂交和流式细胞术检测模型中caspase 3基因表达和神经元凋亡情况.结果:成功克隆获得大鼠全长caspase 3 cDNA,其序列与文献报道一致.模型组海马神经元癫(癎)样放电3 h后caspase 3基因表达开始增多,6 h后凋亡细胞开始明显增加,两者均较对照组明显增加.结论:癫(癎)样放电可能启动神经元caspase 3表达,继而介导神经元凋亡.     

大鼠配对前经历吗啡成瘾及戒断对子代焦虑样行为产生的机制

目的 探讨亲代大鼠配对前经历吗啡成瘾及戒断对子代焦虑样行为产生的机制.方法 选用8周Sprague-Dawley大鼠(雌雄各半)建立吗啡依赖模型,自然戒断21d后配对分组合笼,同时建立生理盐水对照组.子代鼠8周时,雌雄各5只取脑通过Golgi-Cox 染色观察子鼠海马CA1区神经元树突形态情况;另外,子代鼠雌雄再各取5只,深度麻醉后取海马组织作全基因组表达谱分析.结果 吗啡成瘾组子代海马CA1区神经元基树突分枝总长度和基树突分支个数低于对照组子代(P＜0.05);吗啡成瘾组子代与对照组子代相比,雄性子代鼠共同表达2倍上调的有663个基因,共同表达2倍下调的有449个基因,雌性子代鼠共同表达2倍上调的有350个基因,共同表达2倍下调的有188个基因;其中与情绪行为调节相关的基因有:5-HT2c受体上调7倍,Igf-2上调7.1倍,reelin表达下调3.3倍.结论 亲代经历吗啡成瘾戒断导致子代海马CA1区神经元树突发育不良,5-HT2c、Igf-2、reelin等基因表达异常,这可能是子代焦虑样行为产生的原因.     

糖尿病性勃起功能障碍大鼠的基因表达谱芯片数据分析

目的研究糖尿病性勃起功能障碍大鼠基因表达谱芯片数据,应用生物信息学方法挖掘与糖尿病性勃起功能障碍相关的基因,初步探讨其功能及相应分子机制。方法应用GeneSifter在线分析软件对5个正常大鼠阴茎组织和5个糖尿病性勃起功能障碍大鼠阴茎组织基因芯片数据进行分析。结果筛选得到661个差异表达基因,其中280个上调,381个下调。其中kruppel-like factor 5(klf5)基因上调表达4.01倍,ceruloplasmin(cp)基因上调表达5.14倍,collagen,type Ⅺ,alpha1基因下调表达5.84倍,collagen,type Ⅰ,alpha1下调表达5.77倍。661个差异表达基因的功能涉及糖蛋白生物合成、胶原纤维组建、血管发生过程、脂质代谢、细胞增殖等多种重要的生物学过程,以及脂肪酸代谢、神经变性机能紊乱、细胞外基质受体相互作用等信号通路。结论应用生物信息学方法分析糖尿病性勃起功能障碍大鼠基因表达谱发现上调的klf5、cp基因和下调的collagen基因在糖尿病性勃起功能障碍的发病机理中起重要作用,生物信息学方法可为糖尿病性勃起功能障碍发病机制的研究开辟新思路。     

针刺心经、小肠经干预急性心肌缺血下丘脑基因表达

目的:探讨下丘脑在电针干预急性心肌缺血中的作用。方法:将SD大鼠随机分为模型组、肺经组、心经组和小肠经组。采用结扎冠状动脉前降支法复制大鼠急性心肌梗死模型。比较肺经组与模型组、心经组与模型组、小肠经组与模型组的下丘脑基因表达谱变化。结果:与模型组比较,肺经组中大于2倍的差异表达基因(包括EST)有26个下调和57个上调,心经组中有34个下调和190个上调,小肠经组中有97个下调和122个上调。就大于2倍的表达基因而言,仅仅在心经组和小肠经组出现促甲状腺激素释放激素(Trh)基因表达的明显上调和促肾上腺皮质激素释放激素(Crh)基因表达的明显下调。结论:下丘脑差异表达基因特别是Trh和Crh基因表达,可能参与了电针心经、小肠经对急性心肌缺血的保护作用。     

当归对原发性高血压大鼠脑组织基因表达谱的影响

目的探讨当归对原发性高血压大鼠(SHR)大鼠脑组织基因表达谱的影响。方法将SHR分为当归组、模型对照组,并将同周龄的Wistar大鼠作为正常对照组。测定用药前后不同模型组尾动脉收缩压,分组给药4周后,取大鼠脑组织,提取RNA送博淼生物科技(北京)有限公司进行表达谱的测定。结果当归可降低SHR的血压水平。当归组与模型组比较,当归组的变化基因有326个,其中184个基因表达上调,142个基因表达下调:信号值变化幅度2倍以上的差异表达基因85个,上调的基因53个,下调的基因32个,在85个差异表达基因中,已知生物学过程的基因有38个(上调23个,下调15个),主要与信号传导途径的基因相关,另外也与代谢、转录及蛋白质结合等基因相关。结论当归具有一定的降压作用,并对SHR脑组织基因表达谱产生影响,可能主要通过信号传导途径发挥作用。     

基因芯片技术筛选大鼠ARDS差异表达基因的研究

目的 用全基因表达谱芯片技术筛选大鼠急性呼吸窘迫综合征(ARDS)的差异表达基因并对其进行分析,以进一步阐明ARDS发病的分子机制.方法 分别从正常和ARDS大鼠肺组织中提取总RNA,分离纯化mRNA,经逆转录合成掺入生物素标记的cDNA探针,然后与基因芯片(涵盖27044转录本,代表22012个基因)杂交,扫描芯片荧光信号图像,用Sam3.0进行统计处理,用博奥生物分子功能注释系统V4.0进行功能分析.随机选择3个差异表达基因,用荧光定量KT-PCR.验证.结果 芯片检测结果显示,在ARDS大鼠肺组织中,表达上调1.5倍以上的基因有73个,下调1.5倍以上的基因有298个.其中,代谢相关基因上调的有1个,下调的有34个;免疫相关基因上调的有3个,下调的有7个;信号传导相关基因上调的有2个,下调的有9个;神经相关的基因上调的有3个,下调的有3个.结论 全基因表达谱芯片技术是筛选AR.DS差异表达基因的有效方法,用该法筛选出一些差异表达基因,为初步阐明ARDS的发病机制奠定了基础.     

利用基因芯片筛选大肠癌中细胞增殖相关基因

目的 用基因芯片技术筛选大肠癌中细胞增殖相关基因. 方法 利用HG-U133 Plus2.0基因芯片.对大肠癌组织和正常组织基因表达谱进行检测,并利用生物信息学方法对检测结果进行分析. 结果 ①癌组织与正常组织比较,表达差异2倍以上基因共有1 200个,其中表达上调的474个,表达下调的726个.②在1 200个基因中,共有127个基因与细胞的增殖功能相关,其中表达上调的66个,表达下调的61个. 结论 127个细胞增殖相关基因可为大肠癌的基因治疗提供靶点.     

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.