2-脱氧葡萄糖对内质网应激介导的宫内窘迫胎鼠脑损伤的保护机制的研究

目的探讨2-脱氧葡萄糖（2-deoxyglucose，2-DG）对内质网应激介导的宫内窘迫胎鼠脑损伤的保护机制。方法制备胎鼠宫内窘迫模型；实验动物随机分为正常组、缺血再灌注组、假手术组及治疗组；取胎鼠脑组织行TUNEL染色，分析神经元凋亡情况；免疫组化法检测海马回CAl区GRP78及caspase-9，-12蛋白的表达；并观察2-DG对上数指标的影响。结果缺血再灌注组再灌注3hTUNEL阳性细胞数目明显增多；至再灌注24h阳性神经元数量达最大值。治疗组海马回CAl区典型TUNEL阳性神经元数量与缺血再灌注组各相应时间点比较明显减少（P〈0．05）。缺血再灌注组GRP78，caspase-9，-12蛋白表达高于正常及假手术组。GRP78于再灌注3h达峰值，6h内无明显变化，以后随时问的延迟逐渐下降；caspase-12在3h内表达急剧上升，以后趋势变缓，至12h达高峰；caspase-9则于3h内无明显变化，之后迅速增加，至12h达到最大值。治疗组海马回CAl区神经元GRP78蛋白表达均显著高于缺血再灌注组，而神经元凋亡数量及caspase9，-12蛋白表达显著低于缺血再灌注组（P〈0．01）。结论宫内窘迫所致的胎鼠脑组织缺血缺氧启动了内质网应激，内质网相关性死亡途经是胎鼠神经元损伤的又一机制；2-DG通过上调内质网分子伴侣GRP78的表达而对胎鼠缺血缺氧性脑损伤有明确的保护作用。     

caspase-12在胎鼠宫内缺血/再灌注后脑神经元凋亡中的作用

目的 探讨caspase-12与内质网应激介导的宫内缺血/再灌注损害后胎鼠脑组织神经元凋亡的关系.方法 制作胎鼠宫内缺血/再灌注损伤模型.取胎鼠脑组织行TUNEL染色,分析神经元凋亡情况.以免疫组化法检测海马回CA1区caspase-12蛋白的表达情况.结果 缺血15min再灌注3、6、12、24h,海马回CA1区TUNEL阳性神经元数分别为(43.6±11.4)、(64.4±9.3)、(74.2±12.1)和(97.3±8.9)个/mm2;随着再灌注时间的延长,TUNEL阳性神经元数明显增加,于再灌注24h达高峰.与血流未恢复时的(20.2±9.9)个/mm2比较,差异有统计学意义(P＜0.001).缺血15min再灌注3、6、12、24h,海马回CA1区caspase-12的表达分别为0.41±0.12、0.51±0.21、0.59±0.16和0.31±0.09,与对照组的0.01±0.01比较,差异有统计学意义(P＜0.001),表达高峰于再灌注12h出现.结论 caspase-12活化是胎鼠宫内缺血缺氧时内质网应激介导脑神经元凋亡的重要环节.     

多巴胺D1,D2受体激动剂对脑缺血/再灌注损伤的影响

目的 研究多巴胺D1,D2受体激动剂对沙土鼠前脑缺血/再灌注（I/R）海马CA1区神经元凋亡及动物行为学的影响.方法 采用沙土鼠前脑I/R模型,缺血时间5 min.动物分为假手术组、I/R组、SKF-38393组、培高利特组.分别于再灌注1天、3天及7天行开阔法行为学和组织病理学观察以及原位末端标记（TUNEL）法检查CA1区神经元凋亡.结果 行为学检查结果显示,I/R组再灌注各时间点沙土鼠探索活动较假手术组明显活跃（P〈0.05）,培高利特组探索活动较I/R组明显减少（P〈0.05）;病理学及TUNEL结果显示, 培高利特组再灌注3天及7天海马CA1区存活锥体细胞较I/R组增多（P〈0.01）,凋亡细胞较I/R组减少（P〈0.05）.预先应用SKF-38393对以上结果无明显影响.结论 多巴胺D2受体激动剂培高利特能减轻脑I/R海马神经元凋亡及动物行为学异常.     

宫内窘迫胎鼠脑组织葡萄糖调节蛋白78及胱冬酶9,12表达的改变

目的:探讨内质网分子伴侣葡萄糖调节蛋白(GRP)78及凋亡因子胱冬酶(caspase)-9,12在宫内窘迫致胎鼠脑缺血损伤中的作用.方法:制备胎鼠宫内窘迫模型,随机分为正常组(10只胎鼠)、假手术组(50只胎鼠)、缺血再灌注组(50只胎鼠).胎鼠脑组织行TUNEL染色,分析神经元凋亡情况;免疫组化法检测海马回CA1区GRP78及caspase-9,12蛋白的表达.结果:①宫内窘迫缺血再灌注可导致胎鼠脑神经元凋亡,且凋亡数量随再灌注时间延长而增加.②正常组及假手术组仅有GRP78,caspase-9,12蛋白微弱表达;缺血再灌注组GRP78于再灌注3h达峰值,之后逐渐下降;caspase-12于再灌注3h内表达量急剧上升,至12h达高峰;caspase-9则于血流恢复3h之后表达迅速增强,至12h达峰值.结论:宫内窘迫致胎鼠缺血缺氧性脑损伤可能与GRP78表达升高启动内质网自稳调节系统有关;内质网caspase-12凋亡通路的启动可能是脑缺血损伤的又一机制.     

大鼠全脑缺血再灌注损伤后神经元凋亡及认知功能的研究

目的探讨大鼠脑缺血再灌注海马神经元凋亡及认知功能变化。方法雄性SD大鼠60只,随机分成假手术组（SH组）、模型组（IR组）。模型组采用4-VO法建立SD大鼠全脑缺血模型,不同时间点进行水迷宫测试。TUNEL法检测海马CA1区凋亡细胞。结果尼氏染色显示,与假手术组间比较,缺血组海马CA1区神经元数量显著下降（P〈0.01）;TUNEL法检测显示：与假手术组相比,凋亡指数显著增加（P〈0.01）。Morris水迷宫检测,与假手术组相比,模型组大鼠学习阶段寻台时间明显延长,而其记忆测试潜伏期也明显长于假手术组。结论大鼠全脑缺血再灌注损伤后神经元凋亡显著增加,认知功能明显下降。     

低温对脑缺血/再灌注期间沙土鼠海马CA1区延迟性神经元死亡及ATP水平和细胞色素氧化酶活性的影响

目的观察低温对脑缺血/再灌注期间海马ATP水平和细胞色素氧化酶（CCO）活性的影响,以探讨低温减少延迟性神经元死亡（DND）的机制。方法采用沙土鼠前脑缺血/再灌注模型,缺血时间10 min。动物随机分为假手术组、常温组和低温组,3组动物又根据再灌注时间不同进一步分为再灌注6 h、48 h、96 h 3个亚组。所有沙土鼠脑缺血期间脑温均维持（38.0±0.2）℃,再灌注6 h内,低温组脑温维持（30.0±0.2）℃,常温组脑温维持（38.0±0.2）℃。光镜下观察海马CA1区存活锥体细胞数目。ATP、ADP、AMP含量测定采用高效液相紫外检测器法,CCO活性测定采用酶组织化学染色结合计算机图像分析。结果再灌注96 h,低温组海马CA1区存活锥体细胞数目明显多于常温组（P〈0.01）。除再灌注6 h外,低温组海马ATP和腺苷酸池（ATP＋ADP＋AMP）含量均明显高于常温组（P〈0.05）。低温组各再灌注时间点海马CA1区CCO活性均明显高于常温组（P〈0.05）。结论低温可通过保护CCO活性减轻脑缺血/再灌注期间的细胞能量代谢障碍,从而减少DND。     

新生鼠缺氧缺血再灌注后海马c-Fos, c-Jun的表达与神经保护

目的：探讨新生鼠缺氧缺血再灌注后c-fos,c-jun表达与神经元损伤后存活的关系。方法：7日龄SD鼠，经弹性管穿线阻断右颈总动脉3h，予低氧1h；制备成缺氧缺血脑损伤（HIBD）模型。后施以不同时期的再灌注，彩色多普勒监测血流。用免疫组织化学的方法，观察c-fos,c-jun在仔鼠HIBD后不同再灌注时间点海马的表达变化。Thionin染色观察神经元的凋亡。结果：HIBD组右侧海马c-fos,c-jun 6h达高峰，24h稍降，48h又升高，7d后显降低但仍明显高于对照组（P＜0．01）。对照组c-fos,c-jun有极少数表达。Thionin染色发现：CA1区锥体细胞在HIBD再灌注24h后神经元有明显凋亡（P＜0．01），但7d后细胞凋亡无统计学意义（P＞0．05），CA3，DG区神经元也保持形态上的完好。对照组海马各区仅极少数神经元凋亡。结论：c-Fos,c-Jun参与HIBD后海马神经元损伤后修复和存活。     

葛根素对大鼠脑缺血再灌注后梗死范围和锥体细胞的影响

目的 探讨葛根素预处理对大鼠局灶性脑缺血再灌注时梗死范围和锥体细胞的影响.方法 72只SD大鼠随机分为S组（假手术组）、IR组（缺血再灌注组）和P组（葛根素预处理组）,每组24只,依术后处死动物时间的不同每组又分为3个亚组,每亚组8只,同时制作大脑中动脉缺血2h后再灌注损伤的SD大鼠模型.用TTC染色法测定梗死体积,HE染色观察海马区锥体细胞数.结果 S组未见梗死灶,P组脑梗死范围明显比IR组缩小（P＜0.05或P＜0.01）.P24h、P72h组神经元损伤程度较IR24h、IR72h组轻;P24、P72h组存活锥体细胞数明显多于IR24h、IR72h组（P=0.014,P=0.004）.结论 葛根素预处理能减少大鼠局灶性脑缺血后梗死范围和锥体细胞的死亡.     

尼卡地平对脑缺血再灌注后海马CA1区细胞凋亡的影响

目的　研究沙土鼠短暂性脑缺血再灌注后细胞凋亡及尼卡地平处理的影响。方法　实验动物随机分为正常组、假手术组、对照组、尼卡地平处理组。正常组不进行手术操作 ,不进行脑缺血 ;假手术组只进行手术操作 ,不进行脑缺血 ;对照组阻断双侧颈总动脉 15min造成完全性前脑缺血模型 ;尼卡地平处理组在脑缺血前 30min给予腹腔注射尼卡地平 2mg/kg。用TdT介导的原位末端标记 (TUNEL)法来检测死亡神经元的DNA片段。HE染色计数海马CA1区 1mm长度内正常的锥体细胞数。结果　 (1)缺血再灌注后 2～ 4d ,对照组海马CA1区正常锥体细胞数比假手术组明显减少 (P <0 .0 1)。 (2 )缺血再灌注后 2～ 4d ,尼卡地平处理组海马CA1区正常锥体细胞数明显比对照组多 (P <0 .0 1)。 (3)尼卡地平处理组明显减少缺血再灌注后海马CA1区细胞凋亡 (P <0 .0 1)。结论　完全性前脑缺血再灌注后存在细胞凋亡 ;尼卡地平可抑制脑缺血再灌注后海马CA1区细胞凋亡。     

低温对脑缺血再灌注期间海马ATP含量和羟自由基生成的影响

目的研究低温对脑缺血再灌注期间海马ATP和羟自由基（OH·）含量的影响,并探讨二者的变化与延迟性神经元死亡的关系。方法沙土鼠前脑缺血再灌注模型,缺血时间10min。随机将动物分为假手术组、常温组和低温组,后两组又分为再灌注6h、48h、96h三个亚组。每组13只动物,其中5只用于海马组织形态学观察,另8只用于ATP、ADP、AMP和OH-含量测定。ATP、ADP、AMP含量测定采用高效液相紫外检测器法,采用水杨酸捕捉法结合高效液相及电化学检测器法测定OH·含量。结果再灌注96h,常温组海马CA1区存活锥体细胞数目仅为假手术组的5％（P〈0．01）,而低温组为假手术组的45％,明显多于常温组（P〈0．01）。常温组各再灌注时间点海马ATP含量和腺苷酸池（ATP＋ADP＋AMP）含量均明显低于假手术组（P〈0．01）。除再灌注6h外,低温组海马ATP和腺苷酸池含量均明显高于常温组（P〈0．05）。常温组再灌注6h,海马2,3-DHBA含量明显高于假手术组和低温组（P〈0．01或P〈0．05）,但再灌注48h和96h,三组间海马2,3-DHBA含量已无显著性差异。结论延迟性神经元死亡主要与脑缺血后持续的细胞能量代谢障碍有关,而低温可通过减轻细胞能量代谢障碍,起到减少延迟性神经元死亡的作用。     

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.