IS导管血管壁内局部导入c-myc反义寡核苷酸预防PTCA术后再狭窄的实验研究

目的　探讨IS导管局部导入c myc反义寡核苷酸及c myc反义寡核苷酸对血管平滑肌细胞的抑制作用。方法　 2 1只家兔 ,对照组 6只 ,反义寡核苷酸治疗组 15只。用IS导管导入c myc反义寡核苷酸 (1mg)于球囊损伤的髂动脉后 ,动脉血管造影。病理切片 ,图象分析测定新生内膜、中膜、面积和厚度 ,c myc蛋白免疫组化染色。 结果　动脉造影对照组 5 0 %～ 80 %局限狭窄 ,给药组 10 %～ 30 %局限狭窄。光镜观察结果显示 :血管内膜增生面积给药组为 (4.0 4± 1.0 2 )× 10 5μm2 ,对照组为 (7.6 6± 3.7)× 10 5μm ,(P <0 .0 5 ) ;对照组c myc免疫组织化学染色阳性 ,给药组血管c myc免疫组织化学染色显示弱阳性或阴性。结论　IS导管可在体局部导入反义寡核苷酸于动脉损伤靶点 ;c myc反义寡核苷酸可抑制SMCs迁移、增生及新生内膜的形成     

兔髂动脉内膜受损后增生和c-myc基因的表达

研究兔髂动脉内皮剥脱时血管壁细胞的反应特点和 c- myc基因的表达规律。用标准血管成形导管髂动脉球囊损伤 1 2只家兔的右髂总动脉后 ,分别于 3、7、40 d处死动物 ,用透射电镜观察术后 3d平滑肌细胞 ( SMC)超微结构的变化 ,光镜观察术后血管壁细胞的反应特点和 c- myc蛋白表达规律 ,同时对术后 40 d动物行血管造影。结果发现 :术后 3d兔髂总动脉中层 SMC由收缩型转变为分泌型 ,并向内膜下迁移 ,可见管腔内有血栓形成 ,1周时弹力板不完整 ,中膜 SMC排列紊乱 ,术后 40 d增生的 SMC和细胞外基质形成新生内膜 ,血管造影显示管腔狭窄 5 0 %～ 80 % ;球囊损伤段髂总动脉新生内膜 c- myc蛋白免疫组化染色为阳性 ,而对照组为阴性。结论 :球囊损伤髂动脉后可引起 SMC迁移、增生 ,其和细胞外基质一起形成新生内膜 ,管腔狭窄 ;SMC增生和 c- myc基因表达密切相关。     

EFFECT OF STENT ABSORBED c-myc ANTISENSE OLIGODEOXYNUCLEOTIDE ON SMOOTH MUSCLE CELLS APOPTOSIS IN RABBIT CAROTID ARTERY

Ｉｎｔｈｅｐａｓｔｆｅｗ ｙｅａｒｓ ,ｗｅｈａｖｅｗｉｔｎｅｓｓｅｄａｄｒａ ｍａｔｉｃ ｐｒｏｌｉｆｅｒａｔｉｏｎｉｎｔｈｅｕｓｅｏｆｉｎｔｒａｃｏｒｏｎａｒｙｓｔｅｎｔｓ.Ｓｔｅｎｔｓｎｏｗａｃｃｏｕｎｔｆｏｒｏｖｅｒ 70 %ｏｆｐｅｒｃｕｔａ ｎｅｏｕｓｔｒａｎｓｌｕｍｉｎａｌｃｏｒｏｎａｒｙａｎｇｉｏｐｌａｓｔｙ (ＰＴＣＡ ) .Ｈｏｗｅｖｅｒ,ｉｎ ｓｔｅｎｔｒｅｓｔｅｎｏｓｉｓｒｅｍａｉｎｓａｍａｊｏｒｐｒｏｂ ｌｅｍ ,ｏｃｃｕｒｉｎｇｉｎ 2 0 %～ 30 %ｏｆｔｈｅｓｅ ｐｒｏｃ ｅｄｕｒｅｓ[1-3] .Ｉ…     

反义C-myc寡核苷酸对人肝癌细胞SMMC-7721的生长抑制

针对C－mycmRNA第二外显子起站码AUG及下游4个密码子设计合成了反斗、正义及错配寡核苷酸（ODNS），并对合成的ODNs进行了疏代磷酸化修饰。在人肝癌细胞SMMC－7721培养体系中，对反义寡核苷酸（ASODN）抑制细胞增殖的作用进行了研究。发现①与正义和错配ODNs相比较，反义C－mycODN有明显抑制人肝癌细胞SMMC-7721生长的作用;②该生长抑制作用随ASODN剂量增加而增强，随ASODN作用时间而变化;③流式细胞计数仅测定证明反义C－mycODN主要阻断肝癌细胞增殖周期的G1到S期。结果提示：反斗C－mycODN在体外有序列特异性和细胞周期特异性抑制人肝癌细胞的生长作用，且该抑制作用与ASODN作用时间及剂量有关。     

积雪草甙防治PCI术后再狭窄的实验研究

目的探讨积雪草甙防治经皮冠状动脉介入治疗(PCI)术后再狭窄的可行性。方法以球囊剥脱家兔髂动脉内膜后,用积雪草甙治疗4周,于造模及治疗4周后分别采血检测内皮素(ET)、一氧化氮(NO)、C反应蛋白(CRP),并光镜下观察髂动脉的形态学改变。结果家兔髂动脉球囊剥脱后ET、CRP升高,NO下降,与空白对照组相比有显著性差异(P均<0.001);光镜下见模型组血管内皮脱落,内弹力膜断裂,中膜部分断裂,外膜尚完整。积雪草甙治疗后,ET、CRP下降,NO升高,与病理对照组相比有显著性意义(P<0.05),并呈量-效关系;光镜下见内皮细胞覆盖,内膜基本修复,且高剂量组修复优于低剂量组。而病理对照组内皮细胞大部分脱落,内弹力膜断裂,部分伤及中膜,间质细胞大量增生。结论积雪草甙有促进内皮细胞生长、内膜修复的作用,初步提示其具有治疗PCI术后再狭窄的作用。     

ROLE OF ANTISENSE AND DECOY OLIGONUCLEOTIDE OF NF-κB IN VESSEL STENOSIS AND NEOINTIMA FORMATION IN BALLOON-INJURED RAT ARTERY

Objective To examine antisense and decoy oligonucleotides of nuclear factor kappa B in vivo effects on intima proliferation and balloon-injured monocytes chemotactic protein-1 ( MCP-1 ) and extracellular signal regulated kinase-2 ( ERK2 ) expression in the carotid artery of rats. Methods Sprague-Dawley rats underwent balloon-dilation injury of the left carotid artery. Rats are divided into 7 groups ( n = 18 ) and each group includes 6 time points (6 h, 1, 3, 5, 7, 14 d) ( n =3). Uninjured right carotid artery of the same rat was used as controls. Results In model group, sense group and scramble group, vessel intima area , media area and intima/media ratio increased after 5 d and reached the maximum after 7 d. The effect of antisense plus decoy group on intimal hyperplasia was more obvious than that of antisense group and decoy group alone. MCP-1 mRNA expression was increased expression continuously at 3, 5 and 7 d and decreased at 14 d. Compared with model group, sense group and scramble group, antisense group, decoy group and antisense plus decoy group had lowered MCP-1 mRNA expression in each time point ( P ＜ 0. 05 ). NF-κB p65 was dispersed positive stain 6 h after injury and increased after 1 d and peaked at 7 d, but the protein expression was weak at 14 d. ERK2 protein synthesis increased at 1 d and reached the peak at 7 d, while protein expression after 14 d was similar to that at 7 d. Treatment of antisense group,decoy group and antisense plus decoy group inhibited protein synthesis more significantly than those of model group,sense group and scramble group ( P ＜ 0. 05 ). Conclusion NF-κB modulates genes expression and protein synthesis of MCP-1 and ERK2. Celluar proliferation in vessel wall was dynamically changed after balloon angioplasty injury. Antisense and decoy oligonucleotide of NF-κB by local lipofectamine transfer inhibit NF-κB activating gene modulation and neointimal hyperplasia.     

血管回声跟踪技术在评价绝经期女性颈动脉弹性中的应用

目的应用血管回声跟踪技术评价性激素治疗(HT)后血管弹性的变化情况。方法自然绝经1年以上,年龄50～60岁,诊断为更年期综合征者60例,根据是否进行HT分为:激素组30例,坚持HT 1年以上。对照组30例,未进行激素治疗,进行一般对症治疗。应用E-Tracking技术检测颈总动脉的弹性参数,包括弹性模量(Eρ)、僵硬度(β)、动脉顺应性(AC)、单点脉搏波速度(PWVβ)和膨大指数(AI),并将雌二醇(E2)、内皮依赖性舒张功能(EDD)和内膜中层厚度(IMT)与这些弹性参数进行相关分析。结果激素组β、Eρ、PWVβ明显降低,AC升高(P<0.01)。E2与β、Eρ、PWVβ呈负相关(r分别为-0.607、-0.573、-0.574、P均<0.01);而与AC呈正相关(r=0.574,P<0.01)。EDD与β、Eρ、PWVβ呈负相关(r分别为-0.521、-0.411、-0.456,P均<0.01);而与AC呈正相关(r=0.443,P<0.01)。IMT与β、Eρ、PWVβ呈正相关(r分别为0.553、0.444、0.529,P均<0.01);而与AC呈负相关(r=-0.400,P<0.01)。结论绝经期女性随着雌激素水平的下降,动脉僵硬度增加,激素治疗可使动脉弹性得到一定程度的改善。     

正常青春期睾丸的彩色多普勒超声研究

目的　应用彩色多普勒血流显像 (CDFI)对正常青春期睾丸各血管的解剖及血流动力学进行研究 ,建立正常青春期睾丸动脉血流参数的正常值范围。方法　CDFI观察 181例正常青春期青少年的睾丸及睾丸动脉 (TA)、睾丸包膜动脉 (CA)、睾丸内动脉 (ITA)、精索外动脉的走行及分布 ;测量时间平均峰值流速 (TAMX)、收缩期峰值流速 (PSV)、舒张末期流速 (EDV)、搏动指数 (PI)和阻力指数 (RI)。结果　TA、CA、ITA显示率均为 10 0 % ,精索外动脉显示率为 98.6 %。建立了青春期这 4支动脉血管各血流参数的正常值范围。PSV、PI、RI均按TA→CA→ITA的顺序 ,呈不同程度的降低。结论　CDFI可以准确可靠地显示正常青春期睾丸的血管解剖及其血流变化特征 ,有助于青春期青少年睾丸疾病的诊断和鉴别诊断     

TREATMENT OF CANINE ACUTE LIVER FAILURE WITH MODIFIED EXTRACORPOREAL PIGLIVER PERFUSION

Acuteliverfailure(ALF)isaseveresymptomcausedbykindsofreasons ,suchaschemicalandvirologicalones.Becauseofthelife threatingcom plicationsofacuteliverfailure ,75 %of patientswilldiewithinafewdaysofonset.TherewasnosatisfactorytreatmentforALFexceptlivertrans plantation .Althoughlivertransplantationissuccess ful,withasurvivalraterangingfrom 6 0 %to80 % [1] .Thereisashortageofavailabledonoror gans.Onlyalittleamountofpatientsreceivedlivertransplantation[2 ] .Sothereisagreatneedforanef fectiveliver…     

反义HSP70真核表达载体的构建和在人喉癌细胞的表达

目的　进行促凋亡治疗喉癌实验 ,构建和鉴定携带反义HSP70 (heatshockprotein 70 )基因的真核表达载体。方法　将HSP70cDNA反向克隆入 pcDNA 3.1,构建CMV启动子控制的真核表达载体 pcDNA AHSP70 ,用酶切鉴定结果 ;应用该载体转染人喉表皮样癌细胞Hep 2 ,G4 18筛选阳性克隆 ;westernblot和免疫组化检测转染前后瘤细胞的HSP70的表达。检测转染前后瘤细胞的生物学性状。结果　获得 pcDNA AHSP70真核表达载体 ,HSP70反义RNA阻断了Hep 2细胞的HSP70的表达 ,经westernblotting和免疫组化证实 ,实验组瘤细胞不能表达或低表达HSP70 ,而对照和空载体组高表达HSP70。转染反义HSP70的真核表达载体的Hep 2细胞比空载体组和对照组的细胞生长缓慢 ,细胞周期可见实验组出现亚二倍凋亡峰 ,而空载体组没有。结论　本研究成功构建了反义HSP70真核表达载体并在人喉癌细胞得以表达。     

EXPRESSION OF MATRIX METALLOPROTEINASE-9 IN HUMAN ABDOMINAL AORTIC ANEURYSMAL TISSUES

Objective To study the effects of MMP-9 （Matrix Metalloproteinase-9, MMP-9） in the pathogenesis of abdominal aortic aneurysms （AAAs） by localizing the expression of MMP-9 in the aneurysmal tissues. Methods By , the frozen sections （5μm） with aneurysmal tissues （n = 10） were incubated with MMP-9 antibody-added agents, then the sections were stained and observed under the microscope to localize the expression of MMP-9, which displayed a brown precipitate within the arterial walls. The normal arterial wall tissues（n= 10） and the diseased arterial wall tissues from the arterial occlusive diseases （AODs） （n= 15） were also immunized exactly the same way as control. Results A quantity of positive granules which appeared within the aortic media showed the strong expression of MMP-9 in the AAAs, with the positive rate reaching 95 % （19/20）, while no expression of MMP- 9 was observed in the normal artery. However, the scattered distributed positive granules were seen within the arterial wall of some cases of the AOI）s, implying the weak positive expression of MMP-9 in this disease with the positive rate of 26.7% （4/15）. There was a significant difference of the expression of MMP-9 within the arterial wall between the AAAs and AODs（P〈0.01 ）. Conclusion High expression of MMP-9 within the aortic media faciliatates the degradation of collagen and elustin fibres and subsequent dilation of the aortic artery , thus playing an important role in the pathogenesis of AAAs. To refrain MMP-9 from enhanced expressing within the aortic wall is of clinical significance in the prevention and treatment of AAAs.     

延髓动脉的显微解剖观察

应用显微解剖方法观察了30例儿童脑延髓外部各群动脉的组成,并用透明法在立体解剖显微镜下观察了8例延髓内部各群动脉的形态特征、供应范围及其与外部动脉群的关系。延髓外部动脉分为前正中群、前外侧群、外侧群和后群。外部动脉穿入脑实质延续为内部相应的各群动脉。前正中群动脉供应中缝两侧包括舌下神经核、部分锥体束等结构。前外侧群动脉供应锥体束和下橄榄核。外侧群动脉供应延髓网状结构、脊丘束和迷走神经背核等。后群动脉至延髓上部逐渐消失,主要供应薄、楔束核等。     

ALTERATION IN CONTRACTILE RESPONSE TO NORADRENALINE,5-HYDROXYTRYPTAMINE,SARAFOTOXIN 6c,AND ANGIOTENSINⅡIN RAT MESENTERIC ARTERY DURING ORGAN CULTURE

Objective To compare the vasoconstrictive effects of 9 mediators on fresh and incubated mesenteric arteries of rats. Methods The superior mesenteric artery of rat was removed and the endothelium was denuded. The vessels were cut into 1 mm long cylindrical segments and subjected to organ culture for 24 hours. Fresh or incubated segments were immersed into tissue baths and the concentration-response curves were obtained by cumulative administration of the vasoconstrictors. Results In fresh mesenteric artery, endothelin-1 (ET-1), 5-hydroxytryptamine (5-HT), noradrenaline (NA), 5-carboxamidotryptamine (5-CT), and angiotensinⅡ (AngⅡ) induced potent and sustained constrictions in a concentration-dependent manner. The contraction induced by sarafotoxin 6c (S6c) was weak, while bradykinin (BK), des-Arg-bradykinin (DA-BK), and human urotensinⅡ (hUT-II) showed no detectable contraction. The concentraion-response curves in order of slopes was ET-1, NA, 5-HT, 5-CT, and AngⅡ. The order of the maximum contractions was ET-1>NA=5-HT=5-CT>AngⅡ>S6c. After organ culture, the concentration-response curves induced by S6c, NA, and 5-HT were significantly increased, while that induced by AngⅡ was decreased as comparing to fresh arteries. BK contracted the artery only weakly. Conclusion Organ culture changed the phenotypes towards an increased efficacy of NA, 5-HT, S6c, and a reduced efficacy of AngⅡ, which is in accordance with the results of pharmacological characterization in some human vascular disease.     

降钙素基因相关肽与内皮素受体A在兔实验性蛛网膜下腔出血后基底动脉的表达

目的探讨蛛网膜下腔出血(SAH)后脑血管内皮细胞降钙素基因相关肽(CGRP)及内皮素受体A(ETRA)的表达与迟发性脑血管痉挛(DCVS)之间的关系。方法日本大耳白家兔60只,随机分为SAH模型组25只,盐水组25只,穿刺组5只,正常组5只。采用枕大池二次注血法制作SAH模型,灌注处死后取基底动脉制成切片。进行CGRP及ETRA的免疫组化和HE染色,观察CGRP与ETRA的免疫表达和病理结构变化,并用图像分析方法测量血管周长,使用方差分析进行统计学检验。结果基底动脉周长在SAH后均发生明显缩窄,其中在5d时缩小最强烈。CGRP与ETRA免疫阳性标记颗粒在SAH组的基底动脉组织中均有表达,其动态变化规律为CGRP的表达在1h时最明显,在3、5d依次表达减少,5d达到最低点,7d后开始逐渐升高,10d时表达相对比较明显,但未能达到正常水平,CGRP在穿刺组、正常对照组和盐水组也有散在不规则表达。在正常组、穿刺组和盐水组的基底动脉内ETRA有表达;SAH组基底动脉内ETRA在3d组和5d组表现为高表达,以3d组为强,而其中血管内皮细胞表达最强烈,平滑肌也有表达,10d组的表达仍然较强。结论在SAH后脑血管内皮细胞CGRP和ETRA的表达呈现明显的动态改变,但二者的变化趋势并不同步;CGRP和ETRA在DCVS的发生发展中起重要的作用。     

家兔左心室壁内动脉

本文用透明、软X线造影及组织学切片技术,对100例家兔左心空壁内动脉进行了观察,并与人心左室壁内动脉作了比较讨论。家兔左心室壁内动脉的分布类型在许多方面与人的相似。分枝型动脉(ClassA血管)供应心肌全层,包括心内膜下层;直型动脉(ClassB血管)主要分布到乳头肌、肉柱。乳头肌肉动脉分布可分为节段型、轴型和混合型。室间隔左心室侧壁内动脉分布比较紊乱,无明显规律。在心脏横切面中,毛细血管与心肌纤维方向平行,呈袢状分布。     

Ⅱ型糖尿病病人高血压致病因子临床观察

对27例高血压性Ⅱ型糖尿病患者(组1)之高血压致病因子初步进行临床观察,以无高血压之Ⅱ型糖尿病(组2)46例,高血压病(组3)15例及健康人(组4)15例做为对照。测定项目包括OGTT,胰岛素释放试验,血浆心房利钠多肽(ANP),血管紧张素Ⅰ(ATI),血浆醛固酮(ALD),全血容量,尿蛋白定量等。发现高血压性Ⅱ型糖尿病组血浆胰岛素水平、血钠和全血容量显著高于组2,血浆ANP,水平显著低于组4,但无高血压的Ⅱ型糖尿病组也有类似情况,故钠水潴留并不能解释高血压的发病。与高胰岛素血症有关的其他致病因子还须进一步研究。     

平肝降压胶囊对自发性高血压大鼠血管紧张素Ⅱ及肾素活性的影响

目的观察平肝降压胶囊对自发性高血压大鼠(SHR)血浆、下丘脑肾素活性(RA)和血管紧张素Ⅱ(AngⅡ)水平的影响。方法20只雄性SHR随机分为模型对照组、平肝降压胶囊治疗组,另取10只同龄雄性Wistar-Kyoto大鼠作为正常对照组,用药12周后,采用放免法检测血浆和下丘脑RA及AngⅡ的水平。结果平肝降压胶囊能显著降低SHR血压(P<0.01);模型对照组血浆、下丘脑RA及AngⅡ水平较正常对照组明显升高(P<0.05或P<0.01);给予平肝降压胶囊治疗后血浆、下丘脑RA和AngⅡ的水平均明显降低(P<0.05)。结论平肝降压胶囊可能通过降低血浆、下丘脑RA及AngⅡ水平,从而达到降血压作用。     

K-ras基因点突变的反义寡脱氧核苷酸对人胰腺癌靶基因表达的抑制作用

目的　观察针对于胰腺癌K ras基因点突变的反义寡核苷酸对靶基因表达的抑制作用。方法　脂质体将人工合成的针对于K ras基因第 12位密码子点突变的反义寡核苷酸转染体外培养的人胰腺癌细胞株PC 2 ,用免疫组化和原位杂交技术检测靶基因的表达。结果　转染 4 8h后 ,反义寡核苷酸作用组胰腺癌细胞株PC 2的ras蛋白和K rasmRNA的表达程度较正义组和对照组均明显降低 (P <0 .0 5 )。结论　针对于K ras基因点突变的反义寡核苷酸作用于体外培养的胰腺癌细胞 ,对靶基因的表达有明显的抑制作用     

仿生骨植入钛网管固定修复兔骨缺损

目的探讨MHA-bBMP仿生骨作为替代骨对有限接触式钛网管固定下兔股骨干缺损模型的修复作用。方法48只新西兰大白兔随机分组,实验组:MHA-bBMP仿生骨+钛网管;对照组:自体髂骨移植+钛网管。于股骨干中段截除10 mm,建立兔股骨干缺损模型,将MHA-bBMP仿生骨/自体髂骨植入骨缺损处加有限接触式钛网管固定修复骨缺损。观察兔一般情况,检测血碱性磷酸酶,进行X线片、病理组织、电镜等观察。结果钛网管组织相容性好,固定稳定有效,两组均修复了骨缺损。两组血碱性磷酸酶、X线片表现、病理组织、电镜检查的数据经统计分析无显著性差异。结论有限接触式钛网管对植入物和骨缺损断端起到可靠的固定作用;MHA-bBMP仿生骨与自体髂骨植入相比较,诱导成骨活性相当。