丘脑深部电刺激对脑外伤昏迷大鼠意识状态及前额叶皮质Orexins受体OX1R表达的影响

目的探讨丘脑深部电刺激（DBS）对脑外伤后昏迷大鼠的促醒作用及对Orexins受体1（OX1R）表达的影响。方法将54只SD大鼠随机分为3组:空白对照组（n=18,不做任何处理）;对照组（n=18）和DBS组（n=18）。通过自由落体撞击大鼠脑部建立脑外伤昏迷大鼠模型,造模成功后对照组大鼠丘脑植入电极,但不予DBS,DBS组大鼠丘脑植入电极,并予DBS。用意识状态六级评分法评估大鼠的意识状态和行为学变化,并用Western-blot（WB）、免疫荧光技术检测3组大鼠前额叶皮质（PFC）区OX1R的表达水平。结果 DBS 1 h后,DBS组16只出现翻正反射（16/18）,对照组7只出现翻正反射（7/18）,DBS组意识状态水平好于对照组（U=96.00,P=0.026）。在DBS刺激完成后的12 h,WB、免疫荧光技术检测Orexins受体OX1R的表达水平,DBS组均明显高于对照组（P<0.05）;对照组明显低于空白对照组（P<0.05）。结论脑外伤昏迷大鼠可以通过DBS提高它的意识状态水平,其机制可能与DBS能上调PFC区OX1R的表达水平有关。     

晕船适应与不适应大鼠脑干蛋白质双向电泳图谱的比较与鉴定

目的:建立晕船适应及不适应大鼠脑干总蛋白质的双向电泳图谱,鉴定差异表达蛋白,探讨晕船适应的可能机制.方法:30只大鼠根据模拟晕船刺激后异嗜高岭土量的增减分为不晕组(n=10)、晕船适应组(n=12)和不适应组(n=8),另设未接受模拟晕船刺激的正常大鼠作为空白对照组(n=10).应用双向电泳技术,分离晕船适应组和不适应组大鼠脑干蛋白(n=6),肽指纹图谱鉴定两组间差异表达蛋白质.结果:模拟晕船刺激21 d后,与空白对照组相比,晕船适应组大鼠异嗜高岭土量经历了一个由少到多又逐渐回落的过程,不适应组大鼠异嗜高岭土量则始终维持在一个较高的水平(P＜0.01或P＜0.05).获得了9个晕船适应相关蛋白质:硫氧还蛋白过氧化物酶Ⅰ、硫氧还蛋白过氧化物酶Ⅱ、低分子量神经丝蛋白、泛素羧基末端水解酶PGP9.5和谷氨酰胺合成酶上调;碳酸酐酶Ⅱ、磷酸丙糖异构酶Ⅰ、磷酸甘油酸变位酶B和线粒体电压依赖型阴离子通道下调.结论:大鼠对晕船适应后可诱导脑干蛋白质组表达的改变,主要与能量代谢、神经递质调节和氧化应激有关.     

DBS对海人酸癫痫大鼠突触外GABAA受体亚基的影响

目的研究DBS治疗对海人酸癫痫大鼠大脑中突触外GABAA受体δ亚基分布与表达的影响,为癫痫的发病机制与DBS的治疗机制的研究提供参考。方法雄性SD大鼠24只,随机分为正常对照组、癫痫组、DBS治疗组。通过免疫组织化学方法检测突触外GABAA受体δ亚基在大鼠大脑中的分布与表达量的变化。结合大鼠脑电信号的采集分析以及病理形态学观察,分析DBS在癫痫治疗中的作用以及对突触外GABAA受体δ亚基的影响。结果 DBS治疗组大鼠癫痫发作次数(6.01±4.87,n=8)明显低于癫痫对照(17.31±4.59,n=8),P<0.05;DBS治疗可有效抑制癫痫大鼠海马异常放电;DBS治疗组(463.62±38.41,n=8)大鼠海马神经元坏死及胶质细胞增生程度较癫痫组轻(357.12±20.50,n=8)(P<0.05);正常对照组大鼠脑组织突触外GABA受体δ亚基主要分布在海马(IOD=30.96±2.73,n=8)与丘脑腹外侧核(IOD=32.85±2.35,n=8)。癫痫组大鼠脑组织海马(IOD=12.31±1.72,n=8)及丘脑腹外侧核(IOD=22.39±1.94,n=8)突触外GABA受体δ亚基表达下降。DBS治疗组的丘脑腹外侧核(IOD=35.91±3.97,n=8),海马区(IOD=22.74±2.08,n=8)突触外GABA受体δ亚基表达比癫痫组表达增高(P<0.05)。结论丘脑底核DBS治疗可以通过改变大脑突触外GABA受体δ亚基的表达,抑制癫痫大鼠海马的异常放电,减轻癫痫大鼠海马神经元病理变化,起到对癫痫的治疗作用。     

大鼠脊髓渐进性压迫减压后神经细胞黏附分子的表达

目的:研究大鼠脊髓渐进性压迫减压后修复过程中神经细胞黏附分子(NCAM)的表达情况. 方法: Wistar大鼠51只,分为空白对照组(n=3)和损伤组(n=48). 空白对照组不做任何处理,常规饲养;损伤组用螺钉压迫法造成脊髓渐进性压迫模型,压迫至中度开始减压. 按减压后不同时间点又分为1,3,5,7,14和28 d 6个组,每组动物8只. 用免疫组织化学SABC法测定NCAM在脊髓灰质神经元中的表达,并采用计算机Mias软件进行图像分析. 结果:各组灰度值:空白对照组:245.53±3.47;损伤组减压后1 d组:235.33±6.48;3 d组:218.86±2.67;5 d组:198.39±3.24;7 d组:173.15±5.98;14 d组:190.45±4.93;28 d组:227.68±2.85. 与空白对照组相比脊髓渐进性损伤组各不同时间点组均有NCAM的表达,差异具有统计学意义(P＜0.01),其中以减压后7 d NCAM的表达量最高. 结论: 在脊髓渐进性压迫减压后的修复过程中NCAM均有表达,NCAM表达量的变化可能与脊髓损伤后的修复有关.     

吗啡耐受大鼠脊髓蛋白质组双向电泳图谱的建立与差异蛋白鉴定

目的：建立吗啡耐受大鼠腰段脊髓组织蛋白质组双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图 谱,比较分析吗啡耐受大鼠腰段脊髓组织中蛋白质组的变化和差异表达,鉴定出差异表达的蛋白质点并进行验证。 方法：将16只鞘内置管雄性SD大鼠随机分为吗啡耐受组(MT组,n=8)和生理盐水组(NS组,n=8),取其腰段脊髓组织 蛋白以固相pH梯度等电聚焦(immobilized pH gradients isoelectric focusing,IPGIEF)为第一向,十二烷基硫酸钠-聚丙烯 酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electroporesis,SDS-PAGE)为第二向进行2-DE,应用PDQuest分析 软件对考马斯亮蓝染色的2-DE图谱进行图像分析,对差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(matrixassisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)分析,并利用Mascot查询软件搜索Swiss- Prot数据库进行生物信息学分析。采用蛋白质印迹法验证部分差异蛋白。结果：MT组和NS组大鼠脊髓组织2-DE图谱 中各分离出约1 000个清晰的蛋白质点,其中明显差异表达的蛋白有36个。采用MALDI-TOF-MS分析,并利用Mascot查 询软件搜索Swiss-Prot数据库,共搜索到14个蛋白质,与NS组比较,MT组中表达下调的蛋白质点有2个,表达上调的 蛋白点有12个。采用蛋白质印迹法对其中的NADH脱氢酶及伽玛烯醇化酶蛋白质点进行验证,结果与蛋白质组学结 果一致。结论：初步建立了吗啡耐受大鼠脊髓组织蛋白质组双向电泳图谱,证明吗啡耐受可以引起脊髓中多种蛋白 质的表达改变。     

慢病毒介导大鼠卵巢基因转染的实验研究 *

目的 研究活体大鼠卵巢中慢病毒载体介导的基因转染及表达情况。方法 慢病毒载体(携带GFP的Bcl-2-HIV-I)注射大鼠卵巢实质,分别于注射后1、3、7、14、28天处死大鼠（每个时间点实验组n=2,对照组n=2,空白对照组n=2）,通过 Western-blot 电泳检测报告基因的表达。结果 实验组大鼠可见Bcl-2基因的表达,明显高于对照组和空白对照组Bcl-2基因的表达。 结论 慢病毒能有效的将外源基因导入活体大鼠卵巢而无明显毒副作用。     

中药对哮喘大鼠IL-13 mRNA和TNF-αmRNA表达及IgE含量的影响

目的:探讨中药治疗对哮喘大鼠IL-13 mRNA和TNF-α mRNA表达及IgE含量的影响.方法:Wistar大鼠,随机分为中药治疗组(n=8),空白对照组(n=8),哮喘模型组(n=8),利用卵白蛋白(OVA)及氢氧化铝制作大鼠哮喘模型,在雾化激发期,中药组用杏仁、甘草等中药制成的溶液雾化大鼠,其余组用生理盐水代替. 支气管肺泡灌洗液中细胞分类计数,采用免疫放射法、逆转录聚合酶链反应(RT-PCR)半定量法分别检测各组大鼠血清和肺泡灌洗液(BALF)中免疫球蛋白E(IgE)含量以及肺组织IL-13 mRNA和TNF-α mRNA 表达. 结果:与空白对照组比较,哮喘模型组大鼠IgE含量、EOS计数、IL-13 mRNA和TNF-α mRNA的表达显著升高. 与哮喘模型组比较,中药治疗血清IgE的含量、IL-13 mRNA和TNF-α mRNA的表达明显降低(P<0.05). 结论:中药治疗可抑制IL-13和TNF-α mRNA合成,改善哮喘炎症状态.     

miRNA-183在噪声性耳聋大鼠耳蜗组织中的表达及临床意义

目的:探究噪声性耳聋大鼠耳蜗组织中miRNA-183表达情况,并分析其表达水平变化对噪声性耳聋发生发展的影响.方法:将50只SD大鼠分为两组,其中10只设为空白对照组,另40只建立噪声性耳聋动物模型,并随机分为噪声暴露1 d组(n=10)、噪声暴露7 d组(n=10)、噪声暴露14 d组(n=10)、噪声暴露28 d组...     

氯胺酮诱导大鼠丘脑基因表达谱的改变

目的:用基因芯片研究氯胺酮麻醉大鼠丘脑基因表达谱的变化.方法:SD大鼠4只,分为对照组(n=2)和氯胺酮组(n=2).两组大鼠分别腹腔注射生理盐水或100 mg/kg氯胺酮.1 h后,取丘脑,抽提RNA,以RNA为模板逆转录合成cDNA,以cDNA为模板转录生物素标记的cRNA,cRNA经提纯、片断化后与大鼠神经生物学表达芯片U34杂交.杂交后的芯片用扫描仪检测信号,收集图像,用Microarray Suite Version 5.0软件分析差异表达基因并对其进行功能分析.结果:1323条待测基因中,氯胺酮麻醉后237条基因差异表达,上调表达132条,下调表达105条,其中差异表达超过两倍的为59条.差异性表达的基因功能可分为NMDA受体、离子通道、信号转导、转录因子、细胞因子等.结论:氯胺酮麻醉可以引起大鼠丘脑基因的差异表达.     

游泳运动与药物综合治疗大鼠慢性非细菌性前列腺炎的病理学观察

目的 探讨游泳运动与药物综合治疗大鼠慢性非细菌性前列腺炎(CAP)的病理学改变及影响.方法 健康成年SD雄性大鼠60只,随机分为空白对照组(n=12)、CAP模型对照组(n=12)、运动治疗组(n=12)、药物治疗组(n=12)及运动与药物综合治疗组(n=12).采用消痔灵制备CAP大鼠模型.药物治疗组及综合治疗组给予相应药物灌胃治疗;运动治疗组及综合治疗组采用每天1次,每周6d,持续3周的游泳运动;空白对照组、CAP模型对照组及运动治疗组给予无菌生理盐水灌胃.分别于第4、8、11、14、18、22天取各组大鼠前列腺,计算前列腺指数并进行病理学检查.结果 CAP各治疗组大鼠前列腺指数小于模型对照组(P＜0.05),综合治疗组前列腺指数小于药物治疗组及运动治疗组(P＜0.05).在治疗过程的4～22 d,CAP模型对照组大鼠前列腺组织可见明显的慢性炎症病理学改变;运动治疗组、药物治疗组及综合治疗组大鼠前列腺慢性炎症改变均有不同程度减轻,其中,综合治疗组缓解程度较明显.结论 游泳运动有利于前列腺组织炎性病理学损害的缓解,能够辅助大鼠CAP的治疗,与药物并用综合治疗大鼠CAP的病理学改变更加明显.     

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.