鱼藤酮对SD大鼠脑组织神经元损伤的初步研究

目的通过对鱼藤酮导致慢性中毒SD大鼠脑组织标本的病理学检测,探讨鱼藤酮损伤脑神经元的作用机制。方法将20只雄性SD大鼠随机分为两组,实验组:1.5 mg/(kg·d)的鱼藤酮颈背部皮下注射8周,共10只;对照组:相同体积的葵花油颈背部皮下注射8周,共10只。实验中观察各组大鼠的一般情况并每周进行行为学测试及体重的测量。选取实验组全部存活的SD大鼠脑组织标本,共6个;在对照组中随机选取6个脑组织标本。随后进行HE染色,并用显微镜观察组织的病理变化。结果实验结束时,实验组大鼠共死亡4只,对照组无死亡现象。实验组大鼠在实验中表现出较为明显的毛色泛黄、反应迟钝、精神萎靡、活动减少、进食缓慢等中毒症状。在体重增长速度上与对照组比较,差异有统计学意义(P0.05)。在行为学上,实验组大鼠在网格实验中表现出了较为明显的行为学异常,第7、8周时两组大鼠移动的潜伏时间比较,差异有统计学意义(P0.05)。大鼠脑病理学检测提示实验组大鼠脑干区可见神经元数目减少、核固缩、变性等改变。黑质致密部多巴胺(dopanine,DA)神经元变性。对照组大鼠未见明显异常。结论鱼藤酮导致慢性中毒的SD大鼠脑组织神经元发生损伤,并出现神经系统相关的行为学表现,推测鱼藤酮造成线粒体损伤是导致神经元变性坏死的主要原因。     

鱼藤酮纳米脂质载体及其修饰物在大鼠体内的分布

［摘要］目的: 探讨聚乙二醇(polyethylene glycol,PEG)修饰后的鱼藤酮纳米脂质载体(rotenone loaded nanostructured lipid carriers, R NLC)在大鼠体内的分布。方法: 取SD大鼠123只,其中21只随机分为3组,每组7只,分别皮下注射鱼藤酮溶液、R NLC、PEG修饰的鱼藤酮纳米脂质载体(PEG R NLC),在不同时间点取尾静脉血;其余102只SD大鼠随机分为17组,5组皮下注射鱼藤酮溶液,12组分别皮下注射R NLC(6组)和PEG R NLC(6组),在不同时间点取各种组织;采用HPLC法检测血浆和组织样品中鱼藤酮的含量,观察并比较大鼠体内的分布。 结果： 鱼藤酮溶液在血液中达峰时间(Tmax)为4 h,在各组织分布的曲线下面积(AUC)由高到低依次为肾、肝、脑、脾、心、肺。R NLC和PEG R NLC在血液中的Tmax为8 h,AUC分别为鱼藤酮溶液35.99、42.03倍;R NLC在各组织分布的AUC由高到低分别为脑、肝脏、肾、心、脾、肺;与R NLC相比,PEG R NLC在脑组织(黑质、纹状体)中的分布明显升高,在外周组织中的分布均减少。结论： PEG R NLC能增加鱼藤酮在纹状体和黑质中的分布,进一步增强了R NLC的脑靶向效应。     

银杏叶提取物对大鼠内耳组织及蜗神经核线粒体DNA4834bp缺失突变的影响

目的观察银杏叶提取物对大鼠内耳组织及蜗神经核线粒体(mt)DNA4834bp缺失突变的影响。方法取24月龄SD大鼠20只,随机分为实验组和对照组,各10只,分别用银杏叶提取物100mg/(kg·d)、等体积生理盐水灌胃,3个月后处死。干预前后分别测试两组大鼠ABR,荧光定量PCR检测线粒体DNA4834bp缺失率,并观察其与老年听力损失的关系。结果干预后实验组ABR阈移(3.91±3.73)d B pe SPL,对照组ABR阈移(9.64±2.59)d B pe SPL,两组比较,差异有统计学意义(P0.05)。与对照组比较,实验组蜗神经核、内耳组织线粒体DNA4834bp缺失率降低(0.2466±0.1335比1.0461±0.3423,0.3507±0.158比1.0202±0.2269,P0.05)。结论银杏叶提取物能抑制大鼠听觉器官线粒体DNA4834bp缺失突变,减少mt DNA的氧化损伤,改善线粒体功能,改善听力,从而延缓大鼠老年性耳聋进展。     

银杏叶提取物对内耳组织及蜗神经核线粒体DNA4834bp缺失突变的影响

目的；研究银杏叶提取物对内耳组织及蜗神经核线粒体DNA4834bp缺失突变的影响。方法：取SD老龄大鼠(24月龄)20只,随机分为2组,老年实验组10只,用银杏叶提取物灌胃治疗,另设老年对照组10只,每天等体积的生理盐水灌胃3月后处死,用药干预前后分别测试ABR,采用荧光定量PCR检测两组大鼠线粒体DNA4834bp缺失率差别,观察线粒体DNA4834bp缺失率与老年听力损失关系。结果：老年实验组(银杏叶组)用药干预前后ABR阈移增加了3.91±3.73 dB peSPL,老年对照组(生理盐水组)ABR阈移增加9.64±2.59 dB peSPL,两者ABR阈移有统计学差异(P＜0.05)。老年实验组蜗神经核线粒体DNA4834bp缺失率较对照组降低了3.2倍,老年实验组内耳组织线粒体DNA4834bp较对照组降低了1.9倍(P＜0.01)。结论：银杏叶提取物能抑制大鼠听觉器官线粒体DNA4834bp缺失突变,改善老年性聋听力,提示银杏叶提取物能减少mtDNA的氧化损伤,改善线粒体功能,从而延缓老年性耳聋进展。     

阿拉伯木聚糖对帕金森病大鼠行为学和肠道菌群影响研究

目的 观察阿拉伯木聚糖(arabinoxylans, AX)对颈背部皮下注射鱼藤酮葵花油乳化液致帕金森病(Parkinson’s disease, PD)大鼠的行为学及肠道菌群的影响，探讨其对PD模型大鼠的作用及相关机制。方法 选取40只SPF级健康雄性SD大鼠，空白组和假手术组各8只，其余24只造帕金森模型，并随机分为模型组、美多芭组、AX组，各组大鼠分别给予不同干预。干预14 d后进行行为学实验(悬挂实验)评估各组大鼠行为学变化；免疫组化检测大鼠黑质中酪氨酸羟化酶(tyrosine hydroxylase, TH)及α-突触核蛋白(α-SYNuclein, α-SYN)的表达；肠道菌群16S rRNA高通量测序检测各组大鼠肠道微生物的变化。结果 AX组与模型组相比停留时间长(P<0.05);AX组与模型组相比α-SYN蛋白表达较低，TH蛋白表达含量较高；肠道菌群α多样性分析，与模型组相比，AX组和美多芭组ACE、Chao1、Shannon、Simpson指数升高，菌群多样性较丰富，差异有统计学意义(P<0.05);美多芭组与AX组相比，差异无统计学意义(P>0.0...     

缺氧复合NaCN中毒对大鼠心脏病理和cyt-c活性的影响

目的 研究缺氧复合氰化钠(NaCN)中毒对大鼠心功能的损伤.方法 雄性SD大鼠共108只,随机均分为平原实验组和高原实验组.平原实验组大鼠在常规实验室内进行实验;高原实验组大鼠置4 000m低压舱中预处理3d后开始实验.两组各取6只大鼠麻醉后进行气管和左侧颈总动脉插管,术后静待30min后,背部皮下注射NaCN 3.6mg/kg,以RM 6240生理信号采集处理系统记录平均动脉压(mean arterial blood pressure,mAP)、心电图(ECG);再各取18只大鼠,背部皮下注射NaCN(3.6mg/kg)中毒,不同时相点取大鼠心脏,测定细胞色素C氧化酶(cytochrome C oxidation enzyme,cyt-c)活性.再各取30只大鼠,背部皮下注射氰化钠(3.6mg/kg)中毒后,制作心脏常规光镜切片.结果 与平原实验组比较,高原实验组大鼠NaCN中毒后mAP、ECG和cyt-c活性变化更为明显,较难以恢复正常;心肌组织病理切片也发现缺氧复合NaCN中毒后心肌组织的损伤更严重,且更难于恢复正常.结论 高原缺氧后NaCN中毒会产生复合效应,对大鼠心脏功能的损伤更为严重,需要进一步探索相应的防治措施.     

鱼藤酮致帕金森大鼠的脑黑质中凋亡相关蛋白Bcl-2、Bax表达改变

目的 研究鱼藤酮致帕金森病(PD)大鼠中脑黑质凋亡相关蛋白Bcl-2、Bax表达的改变.方法 Wistar大鼠每日颈背部皮下注射鱼藤酮2 mg(kg·d) (3～6周)造模,依据所建立的评分体系记录动物行为变化,在行为学有记分并停止给鱼藤酮4、10 d时,中脑黑质病理切片免疫组化染色比较黑质区域Bcl-2、Bax的表达. 结果在有行为学记分4 d时,记4分和8分的大鼠中脑黑质Bcl-2表达均显著减少;所有PD大鼠中脑黑质Bax表达均显著增加;Bcl-2/Bax比率均显著减少;有记分4 d时,行为学记分与Bcl-2/Bax比值成负相关性. 结论细胞凋亡参与了鱼藤酮帕金森模型大鼠黑质多巴胺神经细胞的损伤.     

不同剂量鱼藤酮对拟帕金森病模型大鼠行为学及纹状体多巴胺

目的 观察不同剂量鱼藤酮皮下注射对大鼠行为学及脑纹状体多巴胺含量的影响,探讨鱼藤酮拟帕金森病大鼠模型的适宜造模条件.方法 Lewis大鼠分别给予鱼藤酮不同剂量(1.0、1.5和2.0 mg/kg/d)皮下注射共28 d.应用旷场实验和斜板实验分别测定大鼠的运动功能和协调性,高效液相色谱-电化学法检测大鼠纹状体内多巴胺含量.结果 模型组大鼠存活率随鱼藤酮注射剂量的升高呈逐渐下降趋势.鱼藤酮2.0 mg/kg组大鼠体重最先明显降低,随着注射时间的延长,各模型组大鼠均出现显著的体重降低.旷场实验结果显示,各剂量鱼藤酮组大鼠跨格次数和站立次数均明显下降.斜板实验结果显示,鱼藤酮1.5 mg/kg组大鼠在斜板的停留角度较对照组显著减小.鱼藤酮1.5 mg/kg组大鼠脑纹状体内多巴胺含量显著降低.结论 鱼藤酮1.5 mg/kg皮下注射28 d,大鼠出现明显的运动功能障碍和脑内多巴胺含量减少,而死亡率相对较低,因此是建立帕金森病大鼠模型的适宜剂量.     

生理剂量的雄激素对去势小鼠主动脉线粒体DNA缺失突变的影响

目的探讨生理剂量的雄激素对C57BL/6J雄鼠主动脉血管壁组织线粒体DNA缺失突变的影响。方法 24只8周龄C57 BL/6J鼠随机分为3组,正常组(n=8),去势组(n=8),去势+生理剂量睾酮组(n=8,颈背部皮下注射睾酮(1 mg/kg,每3天注射1次);另取8只18月龄的自然衰老组,3个月后测定各组血清睾酮浓度,并提取小鼠主动脉线粒体DNA,用巢氏PCR技术分析线粒体DNA缺失突变率。并将缺失片段经纯化后测序。结果去势组线粒体DNA缺失突变率(36.8475±3.3365%)与正常组(18.1713±2.4317%)相比显著增高,而与自然衰老组(42.3075±3.6556%)相比无统计学差异。去势+T组(23.6488±2.7634%)小鼠的线粒体DNA突变率低于去势组、自然衰老组小鼠,而与正常组之间未见统计学差异。测序显示衰老的主动脉mtDNA存在3713、3864、4236、4415这4种缺失突变,且证实缺失前后有同向重复序列。结论老年小鼠主动脉mtDNA存在多种缺失突变,缺失率高于年轻组,雄激素缺乏小鼠主动脉mtDNA缺失突变率增加,生理剂量的雄激素可减少小鼠主动脉线粒体DNA缺失突变率。     

鱼藤酮致线粒体氧化损伤的作用和机制

鱼藤酮是线粒体呼吸链上复合体I的经典抑制剂之一。大量的研究表明当鱼藤酮作用于活体的动物或细胞 时,可导致线粒体功能紊乱、ROS生成增加,蛋白质、脂质、核酸遭受氧化损伤。以线粒体与ROS的生成、鱼藤酮促 线粒体ROS生成机制、鱼藤酮促DNA损伤机制为中心,阐述鱼藤酮致线粒体氧化损伤的相关作用和机制;同时探讨 鱼藤酮作用下线粒体DNA(mtDNA)的损伤及其可能存在的mtDNA突变。     

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.