依达拉奉对海马神经元缺血缺氧损伤后线粒体膜电位的影响

目的 探讨缺血缺氧损伤后依达拉奉对海马神经元线粒体膜电位的影响.方法 原代培养海马神经元细胞,建立缺血缺氧损伤模型,分为正常细胞组、缺血缺氧组、依达拉奉干预组(分别给予1、10、100、300 μmol/L依达拉奉干预)和阳性参照药维生素C干预组,将各组海马神经元细胞用线粒体膜电位敏感性染料Rhodamine-123 进行染色后,激光共聚焦显微镜分别检测线粒体膜电位.结果 与缺血缺氧损伤组比,加入100 μmol/L和300 μmol/L依达拉奉可以显著提高损伤海马神经元的线粒体膜电位,达到正常神经元线粒体膜电位的82%和87%.100 μmol/L和300 μmol/L依达拉奉组之间对线粒体膜电位的的稳定作用没有统计学差异(P>0.05).结论 缺血缺氧损伤后海马神经元线粒体膜电位下降,依达拉奉能够促进线粒体膜电位的恢复,起到神经保护作用.     

依达拉奉对大鼠局灶性脑缺血再灌注损伤的保护作用

目的探讨自由基清除剂依达拉奉对大鼠脑缺血再灌注损伤后细胞凋亡和脂质过氧化的保护作用。方法用线栓法制备大鼠局灶性脑缺血再灌注损伤模型,TUNEL法检测大鼠局灶脑缺血再灌注损伤及依达拉奉干预后海马CA1区原位凋亡情况,用试剂盒检测脑组织丙二醛（MDA）含量和一氧化氮合酶（NOS）活性。结果缺血再灌注损伤后,大鼠海马CA1区神经元细胞凋亡随时间延长而增加,脂质过氧化产物丙二醛含量和一氧化氮合酶活性增高,依达拉奉干预能显著减轻海马神经元凋亡,降低MDA含量和NOS活性。结论在脑缺血再灌注损伤后,依达拉奉能通过清除自由基而减轻细胞凋亡及脂质过氧化损伤。     

异丙酚对缺氧复氧后培养海马神经元nNOS的影响

目的 观察异丙酚对培养海马神经元缺氧复氧后损伤反应的影响.方法 培养12d海马神经元,随机分为正常对照再培养24h组(Nor组)、缺氧4h后复氧24h组(Ano组)、加异丙酚500μmol/L缺氧4h后复氧24h组(Pro组).采用MTT法测定细胞存活率,免疫组化方法 测定nNOS蛋白含量,应用流式细胞仪测定神经元凋亡率.结果 缺氧后Ano组nNOS蛋白表达及细胞凋亡率明显增加(P＜0.01),加入异丙酚后能减少nNOS蛋白含量(P＜0.01)和细胞凋亡率(P＜0.05),Pro组细胞存活率较Ano组增加(P＜0.05),但与Nor组比仍下降(P＜0.05).结论 异丙酚能提高体外海马神经元的抗缺氧能力,减少nNOS蛋白过度表达和细胞的凋亡.     

依达拉奉预处理离体乳鼠脑皮质细胞对脑缺血再灌注损伤的保护作用

目的:探讨不同浓度依达拉奉预处理离体乳鼠脑皮质细胞对脑缺血-再灌注损伤的保护作用.方法:取出生24 h以内的新生SD乳鼠的脑皮质细胞,体外培养至第7天,随机分为5组:A组(正常对照组);B组(药物损伤组);C1组(50 μmol/L依达拉奉预处理组);C2组(100 μmol/L依达拉奉预处理组);C3组(200 μmol/L依达拉奉预处理组).C1、C2、C3组第7天用依达拉奉预处理,24 h后B、C1、C2、C3组予200 μmol/L谷氨酸损伤0.5 h,所有组换正常培养液继续培养24 h后观察神经细胞存活率(MTT法)、乳酸脱氢酶(LDH)漏出率、细胞凋亡率和HE染色后倒置相差显微镜下细胞形态变化.结果:依达拉奉预处理备组MTT含量不同程度高于损伤组,LDH漏出量和凋亡细胞百分比不同程度低于损伤组,倒置相差显微镜下见预处理组细胞形态受损较药物损伤组轻,3个预处理组以C2、C3组效果明显,但两组差别无统计学意义.结论:采用所测浓度的依达拉奉提前24 h预处理离体幼鼠脑皮质细胞,对脑缺血-再灌注损伤有明显的保护作用.     

依达拉奉对庆大霉素诱导肾小管上皮细胞损伤的保护作用

目的:观察依达拉奉对庆大霉素诱导大鼠肾小管上皮细胞损伤的作用及其机制。方法:传代培养大鼠肾小管上皮细胞(NRK-52E),分为对照组、损伤组(2mg/ml庆大霉素)、治疗组A(2mg/ml庆大霉素+10μmol/ml依达拉奉)、B(2mg/ml庆大霉素+20μmol/ml依达拉奉)和C(2mg/ml庆大霉素+40μmol/ml依达拉奉),观察各组细胞增殖能力、乳酸脱氢酶(LDH)水平、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、细胞凋亡率以及bcl-2和Bax mRNA表达情况。结果:庆大霉素刺激NRK-52E细胞后,细胞增殖能力下降、LDH和MDA含量升高、SOD活性下降、细胞凋亡增加、bcl-2和Bax mRNA表达增多。依达拉奉可以提高细胞增殖能力、降低LDH和MDA含量、增强SOD活性、减少细胞凋亡、上调bcl-2mRNA表达以及下调Bax mRNA表达;这些改变随剂量增加而明显(P均<0.05)。结论:依达拉奉可以减轻庆大霉素诱导的大鼠肾小管上皮细胞损伤,其机制可能与减轻氧化应激、抑制细胞凋亡有关。     

依达拉奉对氧剥夺引起的大鼠肠隐窝上皮细胞损伤的保护效应研究

[目的]研究依达拉奉对氧剥夺导致的IEC -6细胞株损伤的保护效应.[方法]将预先培养好的细胞分为3组:正常组(N组),氧剥夺组(OGD,IR组)和依达拉奉治疗组(OGD+依达拉奉,E组).本研究中所用氧剥夺模型是将细胞株换无糖无血清的培养基后放入含5％ CO2和95％N2的密闭培养箱中2h,后重新复氧复糖.在复氧复糖的同时,将依达拉奉盐溶液( 10 mg/mL)加入细胞培养基中.在复氧复糖2h,收集细胞用DHE荧光探针进行氧自由基(ROS)测定;伤后12 h,富集细胞用于超氧化物歧化酶(SOD)和丙二醛(MDA)的测定;伤后24h收集细胞,用流式细胞仪测定细胞凋亡和Alamar Blue测定细胞活力.[结果]DHE荧光探针结果显示OGD导致的ROS,E组比IR组减少:OGD刺激后,IEC -6细胞内的MDA浓度上升为(1.52±0.21) mmol/mg,而E组的含量为(1.21±0.16) mmol/mg,P＜0.05.OGD后,IEC -6细胞内的SOD活力下降至(7.35±0.84) U/mg,而E组的SOD活力为(8.25±1.12) U/mg,P＜0.05.OGD后细胞活力下降为0.48±0.081,而E组的细胞活力为0.56±0.049,P＜0.01.同时,流式细胞仪的结果显示,依这拉奉抑制了OGD导致的细胞过度凋亡.[结论]依这拉奉可以减轻OGD造成的IEC -6细胞的损伤.     

依达拉奉联合高压氧对CO中毒迟发性脑病小鼠血清MDA水平、SOD活性及海马神经元凋亡的影响

目的观察依达拉奉联合高压氧对急性一氧化碳中毒迟发性脑病小鼠血清丙二醛(MDA)水平、超氧化物歧化酶(SOD)活性及海马CA1区神经元凋亡的影响。方法将72只小鼠随机分为对照组、迟发性脑病(DEACMP)组、生理盐水组、高压氧(HBO)组、依达拉奉组及联合治疗组。使用腹腔注射一氧化碳的方法制备一氧化碳中毒迟发性脑病的模型。染毒后15～28 d生理盐水组腹腔注射生理盐水,HBO组行HBO治疗,依达拉奉组腹腔注射依达拉奉,联合治疗组行HBO加腹腔注射依达拉奉治疗。治疗结束后每组分别检测血清MDA水平、SOD活性及海马CA1区神经元凋亡情况。结果①血清MDA、SOD测定结果:DEACMP组小鼠血清MDA含量高于对照组,SOD活性低于对照组(P<0.05);与DEACMP组相比,除生理盐水组外,余治疗组血清MDA含量降低,SOD活性升高,差异有统计学意义(P<0.05);HBO组及依达拉奉组MDA含量高于联合治疗组,而SOD活性均低于联合治疗组,差异有统计学意义(P<0.05);②海马CA1区神经元凋亡:对照组海马CA1区偶见少量凋亡细胞,除生理盐水组外的各治疗组凋亡指数均低于DEACMP组(P<0.05);HBO组及依达拉奉组凋亡指数高于联合治疗组(P<0.05)。结论依达拉奉与HBO联合治疗可以有效降低急性一氧化碳中毒迟发性脑病小鼠血清MDA水平、提高SOD活性,并可以显著抑制海马CA1区神经元凋亡发生,效果优于单纯HBO或者依达拉奉治疗。     

大鼠原代神经元中依达拉奉对糖氧剥夺损伤的保护及对ROS/TXNIP/NLRP3通路的影响

目的 探讨依达拉奉后处理对神经元糖氧剥夺损伤的保护作用及对活性氧(ROS)/硫氧还蛋内结合蛋白（TXNIP)/NOD样受体相关蛋白3( NOD-like receptor associated protein 3, NLRP3)信号通路的影响。方法 培养新生SD大鼠皮质神经元,经免疫荧光染色鉴定后随机分为神经元组、依达拉奉组、糖氧剥夺组、糖氧剥夺+依达拉奉组4组。各组神经元在完成处理后以CCK-8试剂盒检测神经元活力;以细胞凋亡试剂盒检测神经元凋亡情况;以ELISA试剂盒检测神经元ROS产生情况;以蛋白免疫印迹法检测ROS/TXNIP/NLRP3信号通路相关蛋白的表达水平。结果 神经元缺糖缺氧后细胞活力显著降低,依达拉奉处理后又显著上升,差异有统计学意义（P＜0.01）;糖氧剥夺后,神经元凋亡水平、ROS含量、ROS/TXNIP/NLRP3通路相关IL-1β、TXNIP、IL-18、Caspase-1、ASC和NLRP3蛋白表达水平升高,依达拉奉处理后又显著下降,差异有统计学意义（P＜0.01）。结论 依达拉奉对神经元糖氧剥夺损伤具有保护作用,可能与其对R0S/TXN1P/NLKP3信号通路的抑制有关。     

缺氧预处理对 L02肝细胞Bcl-2、Bax、Fas及FasL表达的影响

目的:研究缺氧预处理对正常肝细胞株L02缺血再灌注损伤的抗凋亡机制.方法:将体外培养的L02细胞分为3组:正常对照组(A组)、缺氧复氧损伤组(B组)、缺氧预处理组(C组).各组细胞缺氧复氧处理1、6、1 2、18、24 h后,使用流式细胞术检测细胞凋亡率,各组细胞分别缺氧复氧、缺氧预处理后12 h检测细胞Bcl 2、Bax、Fas及FasL蛋白表达,同时对细胞结构进行电镜观察.结果:C组和B组比较,细胞凋亡率明显降低(P＜0.05),细胞Bcl-2蛋白表达明显升高(P＜0.05),Bax、Fas及FasL蛋白表达量显著降低(P＜0.05),透射电镜观察到缺氧预处理组细胞结构基本正常.结论:缺氧预处理明显减轻了缺氧复氧所导致的L02肝细胞损伤,降低了细胞凋亡率,缺氧预处理保护机制和Bcl-2、Bax、Fas及FasL蛋白表达有关.     

依达拉奉对脑缺血再灌注大鼠脑组织超氧化物歧化酶、一氧化氮、丙二醛水平的影响

目的探讨依达拉奉对局灶性脑缺血再灌注损伤的影响。方法用线栓法制备大鼠局灶性脑缺血再灌注损伤模型,TUNEL法检测大鼠局灶脑缺血再灌注损伤及依达拉奉干预后海马CA1区原位凋亡情况,用试剂盒检测脑组织丙二醛(MDA)含量和一氧化氮合酶(NOS)活性。结果缺血再灌注损伤后,大鼠海马CA1区神经元细胞凋亡随时间延长而增加,脂质过氧化产物丙二醛含量和一氧化氮合酶活性增高,依达拉奉干预能显著减轻海马神经元凋亡,降低MDA含量和NOS活性。结论在脑缺血再灌注损伤后,依达拉奉能通过清除自由基而减轻细胞凋亡及脂质过氧化损伤。     

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.