一氧化碳中毒大鼠一氧化氮含量及一氧化氮合酶活性变化的实验研究

目的探讨急性一氧化碳（CO）中毒后大鼠脑组织和血清中一氧化氮（N0）含量、一氧化氮合酶（NOS）活性变化。方法将20只健康雄性SD大鼠适应性饲养1周后,随机分为对照组和CO染毒组,每组10只。染毒组按60ml／kg体重腹腔注射一氧化碳,每周一次,连续注射四周,对照组注射等量空气。染毒后检测大鼠脑组织和血清中NO含量、NOS的活性。结果CO染毒后大鼠前皮质、小脑组织和血清中NO含量、NOS活性均比对照组高,差异有统计学意义（P〈0.05）。结论NO、NOS可能参与了CO中毒导致的脑损伤。     

依达拉奉对脑缺血再灌注大鼠血脑屏障通透性及MMP-9表达的影响

[摘要] 目的 观察自由基清除剂依达拉奉对创伤性脑缺血再灌注大鼠血脑屏障(BBB)通透性及脑组织基质金属蛋白酶-9(MMP-9)蛋白表达的影响,探讨自由基清除剂保护血脑屏障的作用机制。方法 采用大鼠右侧大脑中动脉缺血再灌注模型,通过测定渗出血管外的EB（伊蓝）含量来评价BBB通透性的变化,采用免疫组化的方法检测各组大鼠脑MMP-9的表达。结果 脑缺血再灌注后3h脑组织EB渗出量增加,1d时达到高峰,2d以后逐渐减小。MMP-9在缺血再灌注12h起显著显著增加,2d达高峰期,依达拉奉组脑组织EB渗出量及MMP-9表达较模型组降低(P＜0.01)。结论 脑缺血后MMP-9表达增高,BBB通透性增加。自由基清除剂依达拉奉可以保护BBB,其作用可能与抑制MMP-9在损伤脑组织中的表达有关。     

微囊藻毒素-LR对大鼠行为认知能力及血脑屏障通透性的影响

目的:通过研究微囊藻毒素-LR(MC-LR)对大鼠认知能力及血脑屏障通透性的影响,探讨MC-LR对大鼠神经系统的影响.方法:40只SD大鼠随机分为对照(生理盐水)组和MC-LR 0.13、0.5、20 μg·kg-1组(每天灌胃染毒经口注射MC-LR),染毒3个月后进行水迷宫实验和旷场实验,观察MC-LR对大鼠学习记忆能力和行为的影响,并检测大鼠伊文思蓝(EB)渗出量以探讨MC-LR对大鼠血脑屏障通透性的影响.结果:与对照组相比,MC-LR染毒3个月后大鼠的学习记忆能力未见显著性差异.旷场实验中,和对照组相比,20 μg·kg-1染毒组大鼠运动距离和自发活动持续时间两个指标差异具有统计学意义(P＜0.05).脑组织EB含量20 μg·kg-1染毒组明显高于对照组(P＜0.05).结论:MC-LR能够增加大鼠血脑屏障的通透性,并可以造成大鼠自发活动行为的改变.     

NG-硝基-左旋精氨酸甲酯对大鼠局灶性脑缺血再灌注损伤脂质过氧化的影响

目的 观察一氧化氮（NO）含量的变化对大鼠局灶性脑缺血再灌注损伤后脂质过氧化的影响。方法 采用线栓法制作大鼠局灶性脑缺血再灌注损伤模型,应用Griss反应法测定脑组织NO含量变化,微量测定法测定超氧物歧化酶（SOD）活性和脂质过氧化产物丙二醛以及NO合酶（NOS）抑制剂NG-硝基-左旋精氨酸甲酯（L-NAME）对它们的影响。结果 脑缺血1 h再灌注1 h脑组织匀浆中NO2-含量升高,SOD活性降低,丙二醛（MDA）含量明显升高,与假手术组比较P<0.01;L-NAME能部分抑制SOD活性的降低及NO2－、MDA含量的升高,与生理盐水治疗对照组比较,P<0.01。结论 小剂量NOS抑制剂能减轻急性脑缺血再灌注损伤脂质过氧化损伤。     

Nω硝基左旋精氨酸甲酯对大鼠海马CA1区毛细血管的影响

目的 探讨Nω硝基左旋精氨酸甲酯(Nω-nitro-左旋精氨酸methyl ester,L-NAME)对大鼠空间学习、记忆能力和海马CA1区毛细血管的影响.方法 内导管成功埋植后,阻断剂组(L-NAME组)大鼠每日给予5 μL的1μmol/μL L-NAME侧脑室注射,对照组(Sham组)每日给予5μL生理盐水侧脑室注射,持续4周.然后用Morris水迷宫测试大鼠空间学习和记忆能力,用试剂盒检测海马组织中NOS活性及NO含量,用体视学方法测定海马和海马CA1区总体积、海马CA1区毛细血管总长度、总表面积和总体积.结果 水迷宫实验中L-NAME组的潜伏期显著长于Sham组(P =0.006);L-NAME组海马组织中的NOS活性及NO含量均显著低于Sham组(P＜0.01);L-NAME组海马体积、CA1区体积与Sham组比较差异无统计学意义(P＞0.05);L-NAME组CA1区毛细血管总长度、毛细血管总表积、毛细血管总体积均较Sham组显著减少(P＜0.05).结论 L-NAME可以导致大鼠空间学习、记忆能力下降和大鼠海马CA1区毛细血管总长度、总表面积以及总体积减少.     

一氧化氮对高热惊厥大鼠海马血红素氧合酶/一氧化碳体系的影响

目的　研究一氧化氮合酶(NOS)抑制剂Nω硝基L 精氨酸甲酯(L-NAME)对反复高热惊厥(FS)大鼠海马血红素氧合酶1(HO- 1)mRNA和蛋白表达及一氧化碳(CO)含量的影响,以探讨一氧化氮(NO)对HO/CO体系的调节作用。方法　采用热水浴诱导大鼠FS。将发育期大鼠随机分为3组:对照组,FS组和FS+L- NAME组。分光光度计间接测定血浆NO和CO含量;核酸原位杂交法检测海马神经元HO- 1mRNA表达;Western印迹方法检测海马神经元HO -1蛋白含量。结果　与对照组比较,反复FS后海马神经元HO -1mRNA表达明显增高,HO -1蛋白含量升高208% (P<0 .01),血浆CO含量增高94 .57% (P<0. 01)。用L NAME干预FS,血浆NO含量下降13 79% (P<0. 05),海马HO 1mRNA表达明显下降,HO .1蛋白表达下降30 .19% (P<0 .01),血浆CO含量下降16 14% (P<0. 05)。结论　反复FS时,外源性给予NOS抑制剂可降低NO含量,同时引起HO 1的基因表达及CO含量下降。     

急性寒冷暴露对大鼠血脑屏障通透性的影响研究

目的:探讨不同时间急性寒冷暴露对大鼠血脑屏障(BBB)通透性的影响。方法:通过-15℃低温暴露1、2、3h建立大鼠急性寒冷暴露模型,用比色法测定脑组织中伊文思蓝(EB)含量,结合干-湿比重法检测脑组织中含水量变化,探讨急性寒冷暴露对大鼠BBB通透性的影响。结果:寒冷暴露3h后大鼠脑组织水分含量和伊文思蓝含量明显增高,BBB通透性明显增加。结论:脑组织在寒冷暴露下BBB通透性增高,可能是寒冷导致中枢神经系统功能障碍的机制。     

急性肝衰竭小鼠Occludin表达与血脑屏障通透性的关系

目的探讨乙酰氨基酚/扑热息痛(acetam inophen/paracetam ol,APAP)导致的急性肝衰竭(acute liverfailure,ALF)小鼠模型中血脑屏障(blood brain barrier,BBB)通透性与脑组织紧密连接(tight junction,TJ)蛋白O ccludin表达的关系。方法应用APAP复制ALF小鼠模型,检测脑中伊文思蓝(evans blue,EB)的含量反映BBB通透性,电镜下观察TJ的结构变化。应用W estern blotting方法检测脑组织中O ccludin蛋白的表达。结果小鼠脑组织EB含量于2 h开始升高(2.29±0.42)μg/g,6 h达到峰值(3.61±0.33)μg/g,脑组织中O ccludin蛋白表达2 h开始下降,6 h表达最低,同时电镜下观察到TJ结构出现破坏。结论在APAP所致的ALF动物模型中,BBB通透性增加,表明血管源性脑水肿机制参与脑水肿形成,与其相反,O ccludin表达呈下降趋势,提示脑水肿的发生与O ccludin表达下降引起TJ结构破坏有关。     

一氧化氮合酶抑制剂N-硝基-L-精氨酸甲酯对局灶性脑缺血再灌注大鼠缺血半暗带区细胞自噬的影响

目的 探究一氧化氮合酶（nitric oxide synthase,NOS）抑制剂N-硝基-L-精氨酸甲酯（Nω-nitro-L-arginine methyl ester,L-NAME）对大脑中动脉闭塞（middle cerebral artery occlusion,MCAO）模型大鼠再灌注后24 h脑内一氧化氮（nitric oxide,NO）和自噬相关蛋白Beclin1、LC3B表达的影响,探讨L-NAME保护脑缺血再灌注损伤的机制。方法 18只健康雄性SD大鼠采用随机数字表法分为假手术（Sham）组、MCAO组,和MCAO+L-NAME组,每组6只。使用改良线栓法制作大鼠右侧大脑中动脉缺血90 min再灌注模型。术中监测大鼠肛温,使其维持在正常范围。于再灌注后24 h处死大鼠,迅速取脑,用免疫荧光染色检测小鼠脑组织冰冻切片缺血半暗带区Beclin1和LC3B的表达水平,用免疫荧光双标分别将Beclin1和LC3B与NO介导的蛋白质损伤标志物3-硝基酪氨酸（3-nitrotyrosine,3-NT）进行共定位。结果 Sham组大鼠没有3-NT染色阳性细胞,偶见Beclin1和LC3B染色阳性细胞,MCAO组小鼠脑组织缺血半暗带区3-NT、Beclin1、LC3B均大量表达（P<0.05）。与MCAO组相比,给予L-NAME治疗后,缺血再灌注大鼠脑组织半暗带区3-NT、Beclin1、LC3B表达均明显减少（P<0.05）。缺血再灌注小鼠脑组织半暗带区,Beclin1和LC3B分别与3-NT免疫荧光染色共定位。结论 L-NAME通过抑制NOS活性,减少NO的产生,直接避免神经元的氧化应激损伤;同时,氧化应激损伤的减弱也避免了自噬的进一步激活,起到了间接的神经保护作用。     

Effect of low-frequency ultrasound therapy on oral mucosal ulcer healing in SD rats

目的 探讨低频超声对大鼠实验性口腔黏膜溃疡愈合时间及溃疡组织中一氧化氮(NO)和一氧化氮合酶(NOS)含量的影响.方法 健康Sprague-Dawley(SD)大鼠36只,雌雄各半,完全随机分为6组(n=6),1组设为正常对照组,其余5组利用氧自由基诱导法+物理烧伤法建立SD大鼠颊囊黏膜口腔溃疡动物模型,分别为溃疡组、药物组和低频超声组(1.1、1.58、2.06 W).末次治疗24h后观察溃疡愈合时间,并采用分光光度计测定大鼠口腔颊囊黏膜组织中NO、NOS含量.结果 大鼠溃疡病理特点与人类口腔溃疡病理特点基本相似.低频超声组较溃疡组及药物组的溃疡愈合时间短.NO活性测定结果显示,低频超声组1.1、1.58、2.06W[分别为(9.86±1.69)、(9.42±1.34)、(8.51±1.26) μmol/g]较溃疡组[ (23.85±4.81) μml/g]显著降低(P＜0.05),与正常对照组(10.88±1.68)μmol/g比较无统计学差异(P＞0.05)；NOS活性检测结果同样表明,低频超声组1.1、1.58、2.06 W[分别为(3.89±0.72)、(4.35 ±0.53)、(4.74±0.81) μmol/g]较溃疡组[(6.48±0.77) μmol/g]显著降低(P＜0.05),与正常对照组(3.83±0.99)μmoL/g比较无统计学差异(P＞0.05).结论 适合剂量参数的脉冲低频超声能有效降低溃疡组织中的NO、NOS活性含量,对溃疡的愈合有一定的促进作用.     

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.