基因芯片技术分析同种异基因大鼠心脏移植急性排斥反应的相关基因

目的:运用基因芯片技术分析同种异基因大鼠心脏移植后急性排斥反应的基因表达谱. 方法:建立大鼠颈部心脏移植模型,以同种同基因和异基因移植动物作为对照,术后5 d移植心脏中抽提总RNA,纯化后的mRNA进行逆转录制备杂交探针,应用含有4 096个靶基因的表达谱芯片对两组移植心脏组织进行差异表达谱分析.结果:在同种异基因心脏移植术后差异表达基因共有210条(下调96条,上调114条);其中已知基因有33条(下调13条,上调20条);未知基因177条(下调83条,上调94条),其中有15条已知基因(上调2条,下调13条)尚未见报道.结论:运用基因芯片技术研究同种异基因心脏移植术后急性排斥相关基因,可为进一步研究其发病机制和治疗提供新思路.     

IL-17在大鼠心脏移植急性排斥反应中的作用

目的 观察白细胞介素17(IL-17)在大鼠心脏移植急性排斥反应中含量的变化及其在排斥反应早期诊断中的作用.方法 采用改良心脏移植术式建立大鼠移植模型.实验组取15只SD大鼠的心脏移植于Wistar 大鼠,对照组取15 只Wistar大鼠的心脏移植于Wistar大鼠.实验组于术后第1、3天和发生急性排斥反应时处死大鼠取血和供体心脏,对照组于术后第1、3、8天获取标本(每次5只).应用酶联免疫吸附实验方法检测血清中IL-17的含量,心脏经过固定和染色后置显微镜下观察心肌组织.结果 实验组急性排斥反应平均发生时间为(7.8±2.9)d.实验组中术后第3天和排斥反应发生时血清IL-17含量明显高于对照组(t=-3.126、-11.043,P<0.05);排斥反应发生时IL-17的含量较术后第1天和第3天显著增高(F=15.113,q=3.72、5.21,P<0.05). 对照组术后第1、3、8天IL-17差异无统计学意义 (t=-0.307～0.097,P>0.05).实验组排斥反应发生时心脏排斥反应病理分级为 4 级,对照组为 0 级.结论 在大鼠心脏移植急性排斥反应发生前 5 d 血清 IL-17 水平已明显升高,排斥发生时达高峰,监测血清 IL-17 的含量可作为早期诊断急性排斥反应的一种方法.     

云南眼镜蛇毒因子用于预致敏大鼠同种心脏移植的实验研究

目的观察纯化的云南眼镜蛇毒因子(Y-CVF)对预致敏大鼠同种心脏移植急性体液排斥反应的作用。方法BN大鼠到Lewis大鼠连续3次皮肤移植预致敏后行颈部异位心脏移植。将15对大鼠用随机数字法分成2组,实验组(n=8)于心脏移植前24h静脉给予Y-CVF80μg/kg;对照组(n=7)不用Y-CVF。观察移植心的生存时间,移植心停跳后病理学检查排斥类型,免疫组织化学染色观察移植心IgG和补体C3的沉积。结果Lewis大鼠预致敏后抗BN大鼠抗体滴度由0升高至1∶1028~1∶2056。对照组移植心存活时间为12·71h±13·94h,实验组移植心存活时间为99·50h±38·72h,与对照组比较差异有统计学意义(t=5·599,P<0·01)。病理检查结果证实,实验组均未发生急性体液排斥,仅见以大量单核淋巴细胞浸润为特征的急性细胞排斥反应。对照组则见以小血管内血栓形成为特征的急性体液排斥反应。免疫组织化学IgG染色实验组和对照组均为阳性,C3染色对照组为阳性,而实验组为阴性。结论使用Y-CVF可克服预致敏大鼠同种心脏移植急性体液排斥反应的发生。     

趋化因子RANTES和受体CCR5在心脏移植慢性排斥反应中的表达

目的探讨调节活化正常T细胞表达和分泌因子(RANTES)及其受体CCR5在心脏移植慢性排斥反应中的表达及意义。方法健康近交系Sprague-Dawley(SD)大鼠为受者,Wistar大鼠为供者,采用"套管连接技术"建立颈部心脏移植的实验模型。将SD大鼠随机分为3组:A组(n=12)供、受者均未采用任何治疗;B组(n=12)移植手术后使用环孢素A(CsA)10 mg.kg-1治疗受者,术后2～3个月采集移植心脏;C组(n=24)使用供者脾细胞(SPC)及环磷酰胺(CP)预处理受者,其中12只在术后15～60 d采集移植心脏(C1组),另外12只术后至少观察半年(C2组)。采用H-E染色观察移植心脏组织排斥反应病理改变程度,Van Gieson染色观察移植心脏血管病变(CAV)的血管狭窄程度,免疫组化检测RANTES和CCR5的表达水平。结果 SPC和CP预处理后,移植心脏的存活时间明显延长,RANTES和CCR5在冠状血管和心肌组织中呈低水平表达,移植心脏血管病和心肌纤维化明显减轻;而在急性排斥反应和CsA治疗的移植心脏组织中,RANTES和CCR5的表达水平明显增强,差异有统计学意义(P<0.05)。结论移植心脏组织中RANTES和CCR5的表达水平与移植排斥反应的程度密切相关。     

IDO基因转染延长同种异基因大鼠移植心脏的存活

目的:了解逆转录基因载体介导的吲哚胺-2,3-加双氧酶(IDO)基因转染能否延长同种异基因大鼠移植心脏的存活时间及机理。方法:把同种异基因大鼠心脏移植模型(Lewis→BN)分成4组,A组为对照组,同种异基因大鼠心脏移植(Lewis→BN);B组:IDO基因转染后的供体心脏移植给受体大鼠;C组:空载体转染供体心脏,再移植给受体大鼠;D组:同种异基因大鼠心脏移植 CsA组(CsA5mg/kg/d×7天)。观察记录移植心脏的存活时间及术后6天组织病理学检查明确排斥反应的诊断和分级,RT-PCR检测术后6天移植心脏TNF-α的mRNA表达,FCM检测外周血活化的T细胞(CD3 CD25 )、有核细胞CD86 表达等方法探讨其机理。结果:A、B、C、D组移植心脏平均存活时间为7.2±0.45、13.2±2.16、7.1±0.62和15.6±3.21天,其中B、D组术后6天CD3 CD25 、有核细胞CD86 表达、供心组织急性排斥反应的强度和TNF-α的mRNA表达较A、C组显著降低(P<0.05),A、C组具有典型重度急性排斥反应特征而B、D组最轻。结论:逆转录基因载体介导的IDO基因转染能够延长移植心脏存活时间的作用,其机理包括抑制反应性T细胞、抑制有核细胞共刺激信号CD86表达、抑制心肌组织表达TNF-α等。     

青藤碱对心脏移植大鼠急性排斥反应及环氧化酶2活性的影响

目的研究环氧化酶2(COX-2)抑制剂青藤碱在大鼠急性心脏移植物排斥反应模型中的作用。方法进行40次SD→W istar的大鼠腹部心脏移植,移植后大鼠随机分为实验组(20只)和对照组(20只),实验组大鼠给予青藤碱30 mg.kg-1.d-1,对照组给予生理盐水,评价生存时间,检测移植心切片,进行排斥反应的病理分析和评价COX-2活性。结果实验组移植物生存时间12.5 d±2.6 d明显高于对照组生存时间6.8 d±0.5 d(P=0.001),移植后3 d、5 d时,实验组的炎性反应、血管内膜炎症、心肌水肿心肌坏死明显减轻。移植心的细胞凋亡明显减少(P<0.05),移植后5 d COX-2蛋白和mRNA表达减少。结论青藤碱能够延长移植物生存时间,减轻心肌损坏和炎症反应。     

血红素加氧酶-1抗急性血管排斥反应的作用研究

目的探讨血红素加氧酶-1（HO-1）在异种心脏移植急性血管排斥反应中的作用。方法建立豚鼠到SD大鼠异位心脏移植模型,选用血红素分别诱导供体和受体,上调HO-1基因表达;眼睛毒蛇因子（CVF）、环孢素A抑制超急性排斥反应。采用免疫组织化学方法检测心脏组织中HO-1的表达;流式细胞仪检钡4受体血清异种抗体IgM的变化;免疫荧光方法观察异种抗体IgM在血管内膜的沉积情况;TUNEL方法检测心脏组织细胞凋亡情况。结果HO-1表达上调后,能明显延长移植心脏的存活时间,抑制异种抗体表达,减少心肌细胞的凋亡。结论HO-1在异种心脏移植中具有抗急性血管排斥反应的作用,对移植器官起保护作用。     

单核细胞趋化蛋白-1和NF-κB通路在心脏移植排斥反应中的作用

目的 探讨单核细胞趋化蛋白-1(MCP-1)在心脏移植排斥反应中的表达及意义,并研究核因子-κB(NF-κB)通路在其中发挥的作用。方法 近交系SD大鼠为受体,Wistar大鼠为供体,“套管连接技术”行颈部心脏移植术。将SD大鼠随机平均分为4组,急性排斥组：受体未采用任何治疗;CsA组：移植术后环孢霉素A(CsA) 10mg/kg治疗受体,术后60～90d采集移植心脏;免疫耐受组：采用Wistar大鼠脾细胞(SPC)和环磷酰胺(CP)预处理SD大鼠,14d后行颈部心脏移植术;PDTC组：移植术后腹腔注射吡咯二硫氨基甲酸酯(PDTC)100mg/kg,1次/d,连续15d。Masson染色观察移植心脏纤维化程度,免疫组化及Western blotting检测MCP-1的表达。结果 急性排斥组、CsA组、免疫耐受组和PDTC组移植心脏存活时间分别为(6.53±2.48)d、(93.51±20.07)d、 (201.42±40.36)d和(142.37±24.64)d,PDTC组明显高于CsA组和急性排斥组(P＜0.05),且心肌纤维化程度明显减轻。各组MCP-1表达的积分光密度(IOD)值依次为(1.86±0.23)、(1.58±0.16)、(0.57±0.15)及(1.16±0.28),免疫耐受组明显低于其他３组,PDTC组显著低于急性排斥组和CsA组(P＜0.05)。结论 MCP-1的表达与排斥反应程度呈正相关,测定MCP-1的水平可预测移植心脏的功能状况;抑制NF-κB通路可减少MCP-1表达,明显减轻心脏移植排斥反应。     

环孢素对心脏移植大鼠急性排斥反应及血清IDO活性的影响

目的 探讨不同剂量的强效免疫抑制剂环孢素(CsA)对同种异基因心脏移植大鼠急性排斥反应程度和血清吲哚胺2,3双加氧酶(IDO)活性的影响.方法 分别以SD和Wistar大鼠为供、受体,建立同种异基因大鼠颈部异位心脏移植模型.18只受体大鼠随机分为对照组、低剂量CsA组(CsA 2 mg·kg-1·d-1)和高剂量CsA组(CsA 15 mg·kg-1·d-1),每组6只.术后第7天,所有受体大鼠经腹主动脉采集血样后处死,取移植心脏制备组织学检查标本.ELISA法检测血清白介素2(IL-2)和γ干扰素(IFN-γ)水平;高压液相色谱法检测血清犬尿氨酸和色氨酸浓度,计算两者比值并以此估算IDO活性.移植心脏标本常规HE染色,采用Stanford标准进行急性排斥反应程度分级.结果 三组血清IL-2和IFN-γ水平以及IDO活性比较显示,对照组＞低剂量CsA组＞高剂量CsA组,差异均有统计学意义(P＜0.05或P＜0.01).三组移植心脏急性排斥反应程度分级结果显示,对照组＞低剂量CsA组＞高剂量CsA组,差异均有统计学意义(P＜0.01).结论 CsA可明显减轻同种异基因大鼠移植心脏急性排斥反应程度并降低血清IDO活性,两者变化趋势一致且与CsA的使用剂量有关.提示血清IDO活性监测有可能成为一种无创性移植器官急性排斥反应及其程度的预测和评估方法.     

WAY-160279预防大鼠心脏移植急性排斥反应

[目的]研究VLA-4(极晚抗原4)拮抗剂WAY-160279预防大鼠心脏移植急性排斥反应的效果.[方法]BN和LEW大鼠分别做供、受体,建立异位心脏移植急性排斥模型.38只受体大鼠按术后用药不同分为对照组和实验组(3组),实验组术后14 d内口服不同剂量WAY-160279,每日2次.[结果]口服WAY-160279(10 mg·kg-1)不能延长移植心脏存活时间(P=0.896),30mg·kg-1及50 mg·kg-1可明显提高移植物存活时间(P＜0.001),并在一定范围内呈剂量相关性.与对照组相比,30mg·kg-1、50mg·kg-1 WAY-160279均可明显减少移植心脏淋巴细胞浸润,减轻组织充血及水肿程度.[结论]WAY-160279可抑制大鼠心脏移植急性排斥反应,显著延长移植物存活时间,并在一定范围内呈剂量相关性.     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

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.