H2S/CBS体系在大鼠癫痫发病中的变化及意义

目的 探讨大鼠癫痫发病过程中H2S/CBS的变化及意义.方法 雄性SD大鼠随机分为正常对照组(NC组)和癫痫模型组(EP组),每组大鼠又分为2h、24 h、7d和30d 4个时间点,氯化锂-匹罗卡品腹腔注射复制癫痫模型,出现Ⅳ级及以上无明显间隔的癫痫持续状态(status epilepticus,SE)为模型复制成功 ;敏感硫电极法测定大鼠血浆中H2S含量 ;免疫印迹(WB)检测大鼠海马中CBS蛋白的表达情况 ;荧光定量PCR检测海马中的CBS mRNA表达变化.结果 癫痫模型组大鼠癫痫发作急性期(2h和24 h)血浆H2S水平、海马CBS mRNA和蛋白表达水平明显高于正常对照组(P＜0.05) ;随后在7d和30 d时H2S/CBS水平降低,显著低于正常对照组(P＜0.05).结论 大鼠癫痫发作急性期H2S和CBS表达水平升高,慢性自发性痫性发作期间H2S和CBS表达水平下降,提示H2S/CBS参与癫痫的发病过程.     

海洛因成瘾大鼠硫化氢/胱硫醚β-合酶体系的变化

目的观察海洛因成瘾对大鼠硫化氢(hydrogen sulfide,H2 S)/胱硫醚β-合酶(cystathionine-β-synthase,CBS)体系产生的影响。方法实验大鼠分成健康对照组、heroin组、heroin+硫氢化钠(NaHS)组,每组10只。heroin组、heroin+NaHS组按剂量递增原则皮下注射海洛因9 d,对照组按同样方式注射等量生理盐水,heroin+NaHS组在每次注射海洛因前30 min腹腔注射NaHS,第10天纳洛酮腹腔注射观察戒断症状。采用亚甲基蓝比色法测定大鼠血浆及海马组织H2S含量,Real-time PCR检测海马组织CBS mRNA表达量。结果 heroin组与健康对照组比较,血浆及海马H2 S含量均明显降低(P<0.05),CBS mRNA表达量显著增高(P<0.05);heroin+NaHS组与健康对照组比较,海马H2S含量明显降低(P<0.05);heroin+NaHS组与heroin组比较,血浆及海马H2S含量明显增高(P<0.05),CBS mRNA表达量明显降低(P<0.05)。结论海洛因依赖可导致大鼠体内H2S水平下降,内源性H2S与CBS之间存在负反馈调节作用,使内源性H2S维持在生理浓度水平。     

硫化氢和胱硫醚β合酶在脑室周围白质软化新生大鼠脑组织中的表达

目的探讨缺氧缺血对3日龄大鼠脑白质细胞硫化氢(H2S)及胱硫醚β合酶(CBS)表达的影响,以及其在3日龄大鼠脑白质损伤(WMD)发病过程中的作用。方法将3日龄大鼠按照随机数字表法分为实验组和对照组,建立新生大鼠WMD模型,分别在缺氧缺血后第12、24、48、72 h和7 d处死大鼠,脑组织取材后行苏木精-伊红(HE)染色,并观察其病理变化。采取免疫组织化学法检测脑组织CBS的表达水平;用分光光度计法测大鼠血浆中H2S水平。结果在新生大鼠脑白质中,缺氧缺血后实验组大鼠脑组织CBS的表达和血浆H2S水平在12 h开始增强,24 h达高峰,7 d仍可见表达;实验组大鼠脑组织CBS的表达和血浆H2S水平在缺氧缺血后12、24、48、72 h和7 d均高于对照组,差异有统计学意义(P<0.05)。结论新生大鼠脑组织CBS和血浆H2S表达水平于缺氧缺血后显著增强,二者可能与新生大鼠WMD的病理生理过程有关。     

糖尿病大鼠肾脏胱硫醚-β-合成酶和胱硫醚-γ-裂解酶mRNA表达初步研究

目的研究糖尿病大鼠肾脏内源性硫化氢(H2S)的两种合成酶胱硫醚-β-合成酶(cystathionine-β-synthase CBS)和胱硫醚-γ-裂解酶(cystathionine-γ-lyase CSE)mRNA表达.方法采用腹腔注射链脲佐菌素诱发糖尿病模型,大鼠随机分为两组:正常对照组(C组)和糖尿病组(D组).建模后第8周,测定24 h尿蛋白、血糖及肌酐清除率[1],应用RT-PCR技术检测大鼠肾皮质CBS、CSE mRNA表达.结果糖尿病组大鼠尿蛋白、血糖及肌酐清除率均较正常对照组明显升高(P＜0.05),但肾皮质CBS、CSE mRNA表达较正常对照组无显著性差异(P＞0.05).结论CBS、CSE/H2S系统不能通过两种合成酶mRNA水平的改变而参与糖尿病肾病中血流动力学紊乱发生,其作用通路中的其他环节改变是否参与糖尿病肾病的发生发展尚需进一步证实.     

胱硫醚-β-合成酶在年轻和老年大鼠海马中的表达分布

目的探讨硫化氢（H2S）合成酶胱硫醚-β-合成酶（CBS）在海马中的分布和表达是否受年龄因素的影响。方法用免疫组化和免疫印迹方法观察年轻（3月龄）和老年（24月龄）大鼠海马中CBS的分布和含量。结果 CBS在年轻和老年大鼠海马中主要分布在颗粒细胞层和锥体细胞层,呈区域性分布,其中CA1区表达最多,CA3区、DG区表达较少;CBS在老年大鼠和年轻大鼠海马中的表达和分布没有变化。结论 CBS在大鼠海马中呈区域性分布,CA1区表达最多,年龄不影响CBS的分布和表达。     

浒苔多糖对非酒精性脂肪肝大鼠脂质代谢及多器官H2S生成的影响

目的 探讨浒苔多糖(EP)对高脂饲料诱导非酒精性脂肪肝(NAFLD)大鼠脂质代谢及多器官内源性硫化氢(H2S)生成的影响.方法 50只成年雌性SD大鼠随机分为5组,每组10只.空白对照组(Normal diet+H2O)、EP对照组[Normal diet+200 mg/(kg·bw·d)EP]、模型组(High-fat diet+ H2O)、EP干预组[High-fat diet+200 mg/(kg·bw·d)EP]、硫氢化钠组[High fat diet+56 μmol/(kg·bw·d)NaHS],持续干预35 d.比较各组大鼠体重、肝脏以及血脂、H2S、CSE、CBS的差异.结果 与模型组比较,EP可显著改善NAFLD大鼠肝细胞脂肪变,降低高脂饲料喂养大鼠血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平(P＜0.05),升高血清H2S水平(P＜0.05),增加多个脏器中胱硫醚-β-合成酶(CBS)/胱硫醚-γ-裂解酶(CSE)活性(P＜0.05).Western blot分析显示,与模型组比较,EP干预后,NAFLD大鼠大脑中CBS和CSE蛋白表达水平、脾脏和肝脏中CBS蛋白表达水平、心脏和肾脏中CSE蛋白表达水平增高(P＜0.05).结论 EP可能通过促进大鼠多器官生成内源性H2S,改善脂质代谢.     

内源性硫化氢在糖尿病大鼠肾脏的变化及作用（英文）

新型内源性气体信号分子-硫化氢(内源性H_2S)具有类似氧化亚氮(nitric oxide,NO)的某些特征,如松弛血管、抑制血管平滑肌细胞增生、调节血管平滑肌细胞张力等作用。本系列研究观察了糖尿病肾病(diabetic nephropathy,DN)大鼠肾脏内源性H2S的变化以及与H2S合成酶-半胱氨酸胱硫醚2β2合成酶(cystathionine 2β2 synthase,CBS)、胱硫醚2γ2裂解酶(cystathionine 2γ2 lyase,CSE)的关系,对DN大鼠肾脏病理改变的影响,对成肌纤维细胞特征性标志物-a平滑肌肌动蛋白(asmooth muscle actin,α-SMA)和细胞外基质成分-纤维粘连蛋白(fibronectin,FN)在大鼠肾组织分布和表达的影响,来探讨内源性H_2S在糖尿病肾病发病机制中的作用。结果显示:①DN大鼠内源性H_2S量明显减少,肾脏CSE的表达亦明显减少。②内源性H_2S的减少与DN大鼠临床表现和肾脏病理改变密切相关。③DN组大鼠肾组织中α-SMA和肾小球基质中FN均增加,分布增多,补充外源性H_2S后,α-SMA和FN明显下降。DN大鼠肾脏内源性H_2S降低。肾脏CSE的减少可能是导致DN大鼠肾脏内源性H_22S降低的直接因素。外源性补充H_2S可抑制糖尿病肾病大鼠肾脏中成肌纤维细胞的生成和细胞外基质的过度积聚。提示,内源性H_2S的减少与糖尿病肾病的发生有关。     

胰岛素样生长因子1对PC12细胞胱硫醚-β-合成酶和硫化氢的影响及机制

\[摘要\]目的观察胰岛素样生长因子1(IGF-1)对大鼠嗜铬细胞瘤细胞(PC12)胱硫醚-β-合成酶(CBS)及硫化氢(H2S)表达的影响以及可能的细胞信号通路机制。方法用不同剂量IGF-1(20、 40、80 ng/ml)作用体外培养的PC12细胞24 h,荧光定量PCR检测CBS基因表达,蛋白质印迹分析检测CBS、细胞外信号调节激酶/丝裂原活化蛋白激酶(ERK/MAPK)相关蛋白表达,敏感硫电极法检测H2S气体含量。之后选取最佳剂量的IGF-1(80 ng/ml) 作用于30 min前已加入ERK/MAPK抑制剂PD98059的PC12细胞再次检测CBS、ERK/MAPK相关蛋白及H2S气体表达水平。结果IGF-1增加CBS的表达和H2S的含量,上调pERK1/2的表达;而PD98059能够抑制IGF-1的上述作用。结论IGF-1及其调控的ERK/MAPK信号通路参与调节CBS及H2S的表达。     

反复热性惊厥前后硫化氢/胱硫醚-β-合成酶体系表达的改变

目的:研究反复热性惊厥(febrile seizures,FS)对发育期大鼠脑内硫化氢(hydrogen sulfide,H2S)/胱硫醚-β-合成酶(cystathionine-β-synthase,CBS)体系表达的影响.方法:采用热水浴诱导大鼠反复热性惊厥.隔日诱导惊厥1次,共诱导10次.发育期大鼠随机分为对照组(N组, n=8)及10次热性处理组(n=22).10次热性处理组依其是否出现惊厥又分为10次高热组(H组, n=9)及10次热性惊厥组(FS组,n=8).10次热性处理组中虽出现惊厥但不足10次者用于其他实验.用亚甲基蓝法测定大鼠血浆H2S含量;用原位杂交和免疫组化法分别观察CBS基因和蛋白表达情况.结果:FS组大鼠血浆H2S含量明显高于对照组和高热组,FS组大鼠海马CBS基因和蛋白表达明显高于对照组和高热组.结论:反复热性惊厥可使大鼠血浆H2S含量升高,并可使大鼠海马CBS基因和蛋白表达增强.     

外源性一氧化碳对脑缺血-再灌注损伤大鼠海马气体信号分子的影响

目的研究外源性一氧化碳（CO）对脑缺血-再灌注损伤大鼠海马气体信号分子的影响,探讨脑保护策略.方法 24只Wister大鼠随机分为四组（n=6）：对照组（Ⅰ组）、脑缺血-再灌注组（Ⅱ组）、脑缺血-再灌注＋低浓度CO组（Ⅲ组）、脑缺血-再灌注＋高浓度CO组（Ⅳ组）.采用四血管阻断方法制作大鼠全脑缺血-再灌注模型,Ⅰ组行假手术,Ⅲ组和Ⅳ组夹闭两侧颈总动脉前30 min腹腔分别注射CO20 mL/kg或40mL/kg,Ⅰ组和Ⅱ组腹腔注射等量生理盐水.缺血20 min再灌注6 h后处死大鼠取海马,检测大鼠海马组织中H2S、NO和CO的量和CBS、HO和iNOS酶活性变化,以及CBS-mRNA、iNOS-mRNA和HO-1-mRNA表达水平.结果Ⅱ组与Ⅰ组相比大鼠海马中H2S、NO和CO的量增高,CBS、HO和iNOS酶活性增加,CBS-mRNA、iNOS-mRNA和HO-1-mRNA表达增高（P〈0.05或P〈0.01）;Ⅲ组与Ⅱ组比大鼠海马中H2S和CO的量增高,NO的量降低,CBS酶活性增加,iNOS和HO酶活性降低,CBS-mRNA表达增加,iNOS-mRNA和HO-1-mRNA表达降低（P〈0.05或P〈0.01）;Ⅳ组与Ⅱ组相比大鼠海马中H2S和CO的量显著增高,NO的量降低,CBS酶活性增加,iNOS和HO酶活性降低,CBS-mRNA表达增加,iNOS-mRNA和HO-1-mRNA表达降低（P〈0.05或P〈0.01）.结论外源性CO能够诱导脑缺血-再灌注后大鼠海马CBS-mRNA的表达,激活CBS,抑制iNOS-mRNA和HO-1-mRNA的表达,抑制iNOS和HO,对HO-1/CO、CBS/H2S和iNOS/NO系统产生调节作用.     

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.