对七氟醚敏感性不同果蝇品系的建立

目的　建立对七氟醚敏感性不同的果蝇品系。方法　将 6～ 8日龄华阴M3 野生型雌、雄果蝇放入特制的玻璃麻醉室内 ,逐渐加大七氟醚浓度实施麻醉 ,用正规法 (Bliss)求出其ED50 值。选出位于 1～ 5百分位最敏感和 95～ 10 0百分位最耐药的雌、雄果蝇作为亲本接种 ,分别产生敏感系第一代 (S1)及耐药系第一代 (R1)。用上述方法测定S1、R1的ED50 值。并将S1代果蝇中位于 1～ 5百分位最敏感雌、雄蝇作为敏感系第二代亲本接种 ,将R1代果蝇中位于 95～ 10 0百分位最耐药雌、雄蝇作为耐药系第二代亲本接种。如此循环筛选接种 ,并测出每代果蝇的ED50 值。敏感系和耐药系均筛选了 11代。结果　两系七氟醚ED50值相差逐渐加大 ,第 11代耐药系的ED50 值是敏感系的 1.5～ 2倍。结论　果蝇对七氟醚的敏感性是可以遗传的。由于雌性和雄性果蝇的七氟醚ED50 值在传代中基本上是同步变化的 ,从遗传学上可以排除隐性纯合的伴X染色体遗传 ,也可以排除显性杂合的伴X染色体遗传。     

基因表达谱芯片对果蝇七氟醚麻醉及敏感相关基因的筛选和分析

目的 采用基因表达谱芯片筛选果蝇七氟醚麻醉及敏感相关基因,并对其基因功能进行分析.方法 以七氟醚敏感、耐药和野生品系黑腹果蝇为实验材料.将果蝇分为两组:七氟醚诱导组,包括三品系果蝇(敏感IS,野生IH,耐药IR);对照组(未经麻醉药干预),包括三品系果蝇(敏感S,野生H,耐药R).采用基因表达谱芯片方法,在各组各品系间进行比较,筛选出果蝇七氟醚麻醉及敏感相关基因.结果 以IH品系为对照,IS品系检测到差异表达基因532个,IR品系171个.其中在敏感品系中参与糖类代谢过程的基因都下调表达,而在耐药品系都上调表达.IS品系对比S品系检测到差异表达基因57个,IH品系对比H品系检测到13个,IR品系对比R品系检测到12个.其中有16个基因在敏感品系与耐药品系表达相反.基因FBgn0010388在各品系七氟醚诱导组都上调表达.结论 三品系果蝇七氟醚诱导后在基因上确实存在差异,且敏感多于耐药品系;其中敏感和耐药品系变化相反的16个基因成为敏感相关基因的可能性较大.敏感品系诱导组比较对照组差异表达基因数最多,野生品系次之,耐药品系最少.FBgn0010388可能与七氟醚麻醉关系密切.     

果蝇吸入麻醉药麻醉作用相关基因的克隆

目的探索吸入麻醉药麻醉作用相关基因或信号传导途径.方法以野生型黑腹果蝇品系及对吸入麻醉药七氟醚敏感和耐药的黑腹果蝇品系为研究模型,通过反转录差异显示PCR方法筛选差异表达的基因片段,经Northem印迹杂交确定阳性基因片段,用cDNA末端快速扩增技术(RACE)克隆阳性基因的全长cDNA并进行序列分析.结果在七氟醚诱导和未诱导的3个品系的果蝇中获得31条差异表达的基因片段;NortherTn印迹杂交确证3条为差异表达的阳性片段;RACE法克隆了2条全长cDNA,一条为1 000 bp的果蝇钙调蛋白基因(No.38),另一条为4 100 bp的功能未知基因(No.45),分别位于Chr.2 R49A-49B和Chr.3 L76B9.结论No.38和No.45基因可能参与吸入麻醉药麻醉作用中信号传导和细胞功能的调节,为开展吸入麻醉药作用的机理研究提供新的途径.     

不同日龄下 3 品系果蝇的七氟醚 ED50和生存力

[目的]了解3个果蝇品系在不同日龄下的七氟醚E队。变化趋势和生存力的差异。[方法]测定HYM3组、F．PS1组和F．PR1组三组果蝇的处女蝇在出生后1—8d、10、12、15、18、22、26、30、34、38、42d的七氟醚ED50，各组内每日龄上的七氟醚ED50均与本组第7日龄果蝇的七氟醚ED50进行t检验。3组雄蝇每组200只分5个培养管饲养，每3日更换1次培养基，并计数生存个体。按寿命表法计算各组果蝇的各项生存力数据。卡方检验比较各组的显著性差异性。[结果]在HYM3组、F．PS1组和F．PR1组中，与7日龄时比较，分别在26日龄以后、15日龄以后和22日龄以后出出了显著性差异和极显著性差异。其七氟醚ED50随日龄增加呈现出下降趋势。组内最小七氟醚ED50值与7日龄时的比值分别为0．70、0．65和0．70。组内变异系数较小的日龄段在HYM3组、F．PS1组和F．PR1组中分别为1～15日龄、1～10日龄和1～12日龄段。3组果蝇经寿命表法得出各项生存力指标无显著性差异。[结论]在3组果蝇品系中，各实验组的七氟醚ED50均随日龄的增长而有一定的降低，但3组改变的趋势基本一致，3组中七氟醚ED50的最小值与7日龄的七氟醚ED50值比值基本一致，分别为0．70、0．65和0．70。3个果蝇品系的各个生存力指标不存在显著性差异。     

抑制性消减杂交筛选与克隆果蝇七氟烷敏感性相关基因

目的 采用抑制性消减杂交(SSH)方法筛选与克隆果蝇七氟烷敏感性相关的基因,以进一步研究吸人麻醉药的作用机制.方法 以对七氟烷敏感和耐药的果蝇品系为研究模型,以野生型黑腹果蝇作为对照,应用SSH方法筛选与吸人麻醉药七氟烷敏感性相关的候选基因.结果 筛选到的差异片段中,在敏感品系中高调的有77个.低调的有56个;在耐药品系中高调的有58个,低调的有52个;在敏感品系中高调而耐药品系中低调的有4个.经过与果蝇基因库比对,确定了其中大部分基因的功能,发现这些基因均非位于Y染色体上;与吸入麻醉药敏感性相关的基因大部分位于第2号染色体上.结论 本实验结果与先前有的学者用遗传学方法定位所得结果基本一致,七氟烷麻醉相关基因定位于2号染色体上,为常染色体显性遗传,并且果蝇对吸入麻醉药的敏感性在雌性与雄性间无差异,为进一步研究吸人麻醉药的作用机制提供了新的思路和方法.     

性别对果蝇吸入麻醉药ED50的影响

目的为了解性别和妊娠对果蝇吸入麻醉药ED50的影响。方法将5～9d龄果蝇分为混合蝇、处女蝇和雄蝇3个处理组,检测七氟醚ED50。在7d龄上进行单因素的重复数据检测,每组检测七氟醚ED50值6次。结果混合蝇、处女蝇、雄蝇的七氟醚ED50分别为0.68±0．04％、0．66％±0．07％、0．64％±0．04％,变异系数（CV）分别为5.7％、11．2％、6．2％。在5、6、8、9d龄上将3组进行涉及性别和日龄两个因素的多因素方差分析,在日龄因素上ED50分别为0．74％±0．03％、0．66％±0．04％、0．71％±0．03％、0.655％±0.05％,性别因素上混合蝇、处女蝇、雄蝇的ED50分别为0．70％±0．06％、0．71％±0.03％、0．66％±0．06％,经多因素方差分析各处理组间无显著性差异,P＞0．05。结论由于性别相差、受孕与否对果蝇吸入麻醉药ED50无明显影响。     

分瓶法测定果蝇吸入麻醉药半数麻醉有效浓度

目的　建立测定果蝇吸入麻醉药半数麻醉有效浓度 (ED50 )的方法学。方法　在 11个 32 5ml的三角烧瓶中各放入 30只雄性果蝇 ,分别加入不同容量的异氟醚标准气 ,使 11个烧瓶中异氟醚浓度分布在零效应浓度与百分之百效应浓度之间。以果蝇失去附壁及运动能力为标准计算各瓶中被麻醉果蝇数 ,用正规法计算出ED50 。结果　 11次实验在 2 5及 5 0min时的量 -效关系曲线均呈S形 ,用各自的对数浓度与概率单位作图都呈直线相关 ,相关系数分别为 0 .972± 0 .0 14和 0 .971± 0 .0 2 5 ,ED50 值分别为 (0 .5 1± 0 .0 6 8) %和 (0 .4 7± 0 .0 85 ) % ,变异系数分别为 13.3%和 18.0 %。结论　本法稳定性较好 ,实用可行     

右美托咪定对全麻气管插管时七氟醚半数有效量值的影响

目的 观察脑电双频谱指数(BIS)达60时右美托咪定对全麻气管插管时七氟醚半数有效量(ED50)值的影响.方法 ASAⅠ或Ⅱ级全麻患者60例,分为两组(n=30):Ⅰ组按序贯法吸入七氟醚全麻诱导,初始呼气末七氟烷浓度为2.0%,相邻浓度间隔比值为1.2,如果5 min内患者的BIS值达60,下一例患者所用七氟醚浓度将下降1个浓度间隔,而5 min内未达到BIS值60者,将七氟醚调整增加1个浓度间隔;Ⅱ组右美托咪定1 μg/(kg·h)静脉泵注10 min后改为0.6 μg/(kg·h)维持,继上述序贯法吸入七氟醚全麻诱导.记录患者清醒到意识消失时间、围插管期平均动脉压(MAP)、心率(HR)、BIS值,统计BIS值达60时两组七氟醚的ED50值.结果 BIS值达60时,Ⅰ组七氟醚的ED50为2.12%(95%置信区间 2.06%～2.57%),高于Ⅱ组的1.61%(95%置信区间 1.52%～1.70%)(P<0.05).Ⅱ组BIS值达60时意识消失时间(40.3±15.4)s,比Ⅰ组的(79.2±17.08)s缩短(P<0.05).Ⅰ组插管后MAP、HR高于插管前(P<0.05),Ⅱ组插管后循环稳定.结论 右美托咪定能够显著地降低全麻气管插管时七氟醚ED50值和血流动力学反应,缩短意识消失时间.     

累积法测定异氟醚对哌库溴铵量效关系的影响

目的应用累积法测定不同质量浓度异氟醚对哌库溴铵量效关系的影响。方法将 30例全麻患者随机均分为 3组 ,A组为对照组 ,B组吸入异氟醚 0 .5肺泡最低有效浓度 (minimumalveolarconcentration ,MAC)、C组吸入异氟醚 1.0MAC ,分别用累积法测定哌库溴铵的半数有效量 (medianeffectivedose,ED50 )和 95 %有效量(95 ?fectivedose ,ED95)。结果与对照值相比较异氟醚 0 .5MAC使哌库溴铵的ED50 值减少 2 5 % ,ED95值减少15 % ;异氟醚 1.0MAC使哌库溴铵的ED50 值减少 6 0 % ,ED95值减少 4 9%。结论异氟醚增强哌库溴铵的肌肉松弛效应 ,异氟醚吸入的质量浓度越高 ,增强哌库溴铵的肌肉松弛效应越明显。     

乳化异氟醚的饱和溶解浓度及在大鼠静脉注射时的半数有效量

目的确定乳化异氟醚静脉注射制剂的安全配制浓度,并测定其在大鼠静脉注射时的麻醉效价.方法以顶空两次平衡法测定20℃时异氟醚在20%和30%Intralipid(R)中的液/气分配系数(λL/G).根据气体物理学原理,计算乳化异氟醚的饱和溶解浓度(Cs).以30%Intralipid(R)为载体,室温下配制浓度为3.1%(v/v)的乳化异氟醚.应用序贯法,以前爪翻正反射消失为判断成功麻醉的指标,测定乳化异氟醚在对成年雌性SD大鼠(体重180～220g)实施单次静脉麻醉诱导时的半数有效量(ED50),并记录大鼠翻正反射消失和恢复时间.结果20℃条件下,异氟醚在20%和30%Intralipid(R)中的λL/G分别为34.90±2.58和51.01±1.55,Cs(95%可信区间)分别为5.64%(5.30%～5.97%)和8.24%(8.04%～8.44%).静脉注射乳化异氟醚后,所有麻醉大鼠的翻正反射均在5秒内消失.乳化异氟醚在大鼠静脉注射时的ED50为(0.073±0.016)ml/kg.ED50测定过程中无动物死亡,麻醉大鼠前爪翻正反射恢复时间为(22±15)s.结论室温下,每100ml异氟醚脂肪乳剂中异氟醚的最大溶解量分别为5.64ml(20%Intralipid(R)为载体)和8.24ml(30%Intralipid(R)为载体).从制剂的安全范围角度考虑,30%Intralipid(R)更适合作为乳化异氟醚的载体制剂.静脉注射乳化异氟醚在大鼠产生可逆性麻醉作用,并且苏醒速度快.     

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.