不同剂量人组织激肽释放酶对兔症状性脑血管痉挛的影响

目的 探讨不同剂量人组织激肽释放酶(HTK)对兔症状性脑血管痉挛的影响.方法 双侧颈总动脉结扎后2周无神经功能障碍的日本大耳白兔40只,体重2.0～2.5 kg,随机分为5组(n=8):对照组(C组)、症状性脑血管痉挛组(V组)和低、中、高剂量HTK组(L组、M组和H组).采用枕大池二次注血法建立兔症状性脑血管痉挛模型,V组于第1次枕大池注血后耳缘静脉注射生理盐水5 ml/d,L组、M组和H组每天分别耳缘静脉注射HTK 0.002 5、0.005 0、0.010 0 PNAU/kg,连续14 d0于枕大池注血前1 d(T_1)及注血后每天评价神经功能并记录进食量,于T_1及枕大池第1次注血后5、7、14 d(T_(2～4))时采用三维CT血管造影测定基底动脉直径,第15天时取基底动脉观察病理学结果.结果 与C组比较,V组低进食量和神经功能障碍发生率升高(P＜0.05);与V组比较,L组、M组和H组低进食量与神经功能障碍发生率降低,T_3时基底动脉直径增加,H组T_4时基底动脉直径增加(P＜0.05);与T_1时比较,V组T_(2,3)时基底动脉直径缩短(P＜0.05),L组、M组和H组T_(2,3)时基底动脉直径差异无统计学意义(P＞0.05),T_4时基底动脉直径增加(P＜0.05);与L组和M组比较,H组T_4时基底动脉直径增加(P＜0.05).结论 耳缘静脉注射HTK(0.002 5～0.010 0 PNAU/kg,连续14 d)可改善兔症状性脑血管痉挛,其效应呈剂量依赖性.     

人组织激肽释放酶6在前列腺癌中的表达和意义

目的初步探讨人组织激肽释放酶6(Kallikrein 6,KLK6)在前列腺癌中的表达和意义。方法搜集徐州医学附属医院泌尿外科2010-06—2013-10行前列腺根治手术患者组织标本共65例,其中40例前列腺癌(Prostate Cancer,PCa),25例为前列腺增生组织,应用免疫组织化学两步法,检测组织中KLK6蛋白表达情况。结果 PCa患者组织中KLK6阳性表达率为75%,前列腺增生(Hyperplasia of prostate,BPH)组织KLK6阳性表达率为24%,两者比较癌组织显著高于增生组织(P0.01);前列腺癌中低危患者KLK6阳性表达率为58.25%,中高危患者为75%,两者比较中高危患者明显高于低危患者(P0.05)。结论 KLK6在前列腺癌组织表达明显高于前列腺增生,中高危患者明显高于低危患者,KLK6基因可能在前列腺癌诊断和预后判断中存在一定价值。     

人组织激肽释放酶12在胃癌中表达的临床意义

目的 探讨人组织激肽释放酶12(KLK12)在胃癌患者外周血血清及肿瘤组织内过度表达的临床意义.方法 收集2008年6月至2009年5月上海交通大学医学院附属仁济医院行胃癌根治术患者的癌组织标本,应用RT-PCR和Western blot检测KLK12在行胃癌根治术患者的胃癌组织、癌旁组织和正常组织的表达情况,检测胃癌患者外周血血清KLK12含量,并以同期非胃癌手术患者外周血血清KLK12的含量作为对照.采用t检验、单因素方差分析.结果 胃癌组外周血KLK12含量为(0.958±0.112)nmol/L,对照组为(0.675±0.101)nmol/L,两组比较,差异有统计学意义(t=13.27,P＜0.05).胃癌低分化者外周血KLK12含量为(1.027±0.165)nmol/L,高分化者为(0.937±0.108)nmol/L.胃癌组织、癌旁组织和正常组织中KLK12 mRNA表达水平分别为39±9、28±6和19±3;蛋白表达水平分别为33±8、30±5和23±4,3者比较,差异有统计学意义(F=3.48,1.48,P＜0.05).结论 KLK12基因在胃癌患者的血清和肿瘤组织中存在过高表达,检测KLK12可能在胃癌诊治中发挥重要作用.     

人组织激肽释放酶在消化系统肿瘤中的表达及预后价值

人组织激肽释放酶（KLKs）属于丝氨酸蛋白酶家族，广泛存在于不同组织和生物液体中，共编码15 个分泌型蛋白KLK1-KLK15。近年来相关研究表明，人组织激肽释放酶与消化系统肿瘤的发生、发展、侵袭和转移密切相关，可以作为消化系统肿瘤标志物应用于疾病的早期疾病筛查、诊断、个性化治疗以及预后判断等临床实践中。本文通过回顾目前临床上常见的五种消化系统肿瘤：食管癌、胃癌、结直肠癌、原发性肝癌及胰腺癌，分析KLKs在消化系统肿瘤中的基因表达及预后评估情况，以期探索更加特异、灵敏的肿瘤生物学标志物来提高消化系统肿瘤的早期诊治和预后判断，为后续临床研究奠定一定的理论基础。     

人组织型激肽释放酶基因对大鼠糖尿病肾病的影响及其机制

目的：探讨人组织型激肽释放酶基因（human tissue kallikrein,HK）对糖尿病肾病（diabetic nephropathy,DN）的影响及其机制。方法：48只雄性Wistar大鼠随机分为普通饲料组（即正常组）和高脂高糖饲料组。高脂高糖饲料组以小剂量链脲佐菌素加高脂高糖饮食诱导2型糖尿病（diabete mellitus,DM）模型,造模后随机分为两组,分别经尾静脉导入重组腺相关病毒（recombinant adeno-associated viruses,rAAV）介导的HK基因（HK组）以及报告基因beta-半乳糖苷酶（beta-galac-tosidase,LacZ）（LacZ组）,观察12周后处死实验动物,留取心、肝、肾等组织。Western blot检测HK在各组织的表达。PAS染色观察肾小球硬化情况,天狼猩红染色观察肾脏纤维化情况。24h尿微量白蛋白（microalbuminuria,mAlb）与尿渗透压检测评定肾功能,免疫组织化学以及Western blot检测肾脏转化生长因子-β1（transforming growth factor-β1,TGF-β1）的表达。结果：rAAV介导的HK可在2型DM大鼠的肝脏、肾脏、心脏表达,而且相较于LacZ组且表达增强;PAS染色显示：LacZ组肾脏出现中度的肾小球系膜细胞及基质增生,血管周围间质细胞浸润,血管壁增厚,以及肾小囊滴状病变和毛细血管纤维帽。另外肾小管大量的空泡变性,肾小管腔内出现较多蛋白质管型,HK组较LacZ组明显好转;天狼星红染色显示：LacZ组肾脏间质、血管壁胶原沉积,肾小球细动脉管壁增厚,而HK组病变较轻（P〈0.05）;24hmAlb排泄量LacZ组比正常组明显增加（P〈0.05）,HK组较LacZ组排泄量显著下降（P〈0.05）。LacZ组与正常组比较,尿渗透压明显下降（P〈0.05）,HK组较LacZ组显著上升（P〈0.05）;TGF-β1主要表达于LacZ组的肾小管上皮细胞,HK组较LacZ组TGF-β1明显下调（P〈0.05）。结论：HK基因明显改善DN,保护肾功能,原因可能与下调肾脏TGF-β1的表达有关。     

组织激肽释放酶基因7在前列腺癌组织中的表达

目的 探讨组织激肽释放酶基因7(KLK7)在不同前列腺组织中的表达情况.方法运用逆转录聚合酶链反应法检测正常前列腺(5例)、良性前列腺增生(BPH)及BPH细胞株(BPH1,13例)、前列腺癌及前列腺癌细胞株(8例)的上皮细胞中KLK7mRNA表达水平;蛋白质印迹法检测不同前列腺组织上皮细胞中KLK7蛋白表达水平;免疫组化分析正常前列腺(20例)、BPH(50例)、前列腺癌(103例)组织中KLK表达水平.根据染色强度分为4个等级(-,+,++,+++)进行半定量分析,染色强度++及+++者判定为阳性.结果 正常组、BPH组和前列腺癌组KLK7 mRNA表达相对值分别为0.59、0.52、0.02,组间比较差异有统计学意义(F=13.03,P＜0.01),前列腺癌上皮中KLK7 mRNA表达下调(P＜0.01),正常前列腺和BPH上皮中KLK7 mRNA表达差异无统计学意义(P＞0.05).KLK7蛋白在正常前列腺、增生前列腺、DU145、LNCaP、PC3、22RV1、BPH细胞株中表达水平相对值分别为0.22、0.40、0.01、0.05、0、0.03、0.14.免疫组化染色结果 显示正常前列腺组织、BPH组织、前列腺癌中KLK7蛋白表达阳性率分别为65.0%(13/20)、76.0%(38/50)、17.5%(18/103),前列腺癌组与前2组比较差异均有统计学意义(P＜0.01),前2组间比较差异无统计学意义(P＞0.05).结论 KLK7在前列腺癌组织中表达下调,提示KLK7在前列癌的发生和进展中可能起一定作用.     

激肽释放酶激肽系统在大鼠阴茎海绵体表达的研究

目的:研究激肽释放酶激肽系统在成年大鼠阴茎海绵体的表达。方法:利用逆转录聚合酶链反应(RTPCR)技术分别检测SD大鼠阴茎和心脏组织中组织型激肽释放酶Ⅰ和激肽B2型受体mRNA的表达情况。结果:大鼠心脏与阴茎海绵体组织中均表达组织型激肽释放酶Ⅰ和激肽B2型受体mRNA,两者有几乎相同的表达水平,差异无显著性(P>0.05)。结论:在生理状态下,激肽释放酶激肽系统存在于阴茎海绵体组织中,而且与心血管系统的表达水平相当。     

人腺体激肽释放酶（hK2）在前列腺癌诊断中的应用

人腺体激肽释放酶（hK2）是一种在诊断前列腺癌中具有非常重要意义的血清标记物。本文主要综述了hK2的结构，及其在前列腺癌中的诊断，病理分级与在评价化学性预防疗效中的应用。     

组织激肽释放酶及其基因与肾脏疾病

组织激肽释放酶是激肽系统的重要组成部分 ,参与一系列肾脏的生理、病理变化 ,与肾脏疾病的发生发展有着密切的联系。本文拟就有关组织激肽释放酶及其基因与肾脏疾病的研究现状及进展作一综述。     

人激肽释放酶对离体动物阴茎海绵体平滑肌的舒张效应

目的探讨人组织激肽释放酶对离体动物阴茎海绵体平滑肌的舒张效应,以初步探讨其对阴茎勃起的调节作用。方法通过离体阴茎海绵体平滑肌肌条实验方法,利用PowerLab4SP微弱生物信号采集系统记录人组织激肽释放酶对离体家兔和大鼠阴茎海绵体平滑肌的舒张效应。结果100mU人组织激肽释放酶可分别使去氧肾上腺素诱导的家兔和大鼠阴茎海绵体平滑肌舒张(65.98±6.98)%和(54.86±9.65)%。结论人组织激肽释放酶可强力地舒张离体家兔和大鼠阴茎海绵体平滑肌,推测其对人阴茎海绵体平滑肌也会有较强的舒张效应,可能是勃起功能障碍药物开发的一个新靶点。     

上胸段硬膜外交感神经阻滞对兔蛛网膜下腔出血诱发脑血管痉挛的影响

目的 探讨上胸段硬膜外交感神经阻滞对兔蛛网膜下腔出血(SAH)诱发脑血管痉挛的影响.方法 日本大耳白兔24只,雌雄不拘,体重2.0～2.6 kg,随机分为3组(n=8).对照组(C组)枕大池注射生理盐水1 ml/kg,硬膜外腔注射生理盐水0.5 ml/2 h;S组枕大池注射自体动脉血1 ml/kg,硬膜外腔注射生理盐水0.5 ml/2 h;H组枕大池注射自体动脉血1 ml/kg,硬膜外腔注射0.5%利多卡因0.5 ml/2 h.记录家兔实验第4、5、6天进食量和神经功能状态,经颅多普勒超声测定实验前(T0)和实验第7天(T1)颈总动脉血流速度[平均血流速度(Vm)、收缩期峰血流速度(Vs)、舒张末血流速度(Vd)]搏动指数(PI)、阻力指数(RI),连续监测心率(HR)、平均动脉压(MAP)、心电图.结果 与C组相比,S组进食量下降和神经功能障碍发生率增加(P＜0.05);与S组相比,H组进食量下降和神经功能障碍发生率减少(P＜0.05);与T0时相比,T1时S组Vs、Vm及Vd升高,PI及RI降低,H组HR及上述指标降低(P＜0.05);与C组相比,T1时S组Vs、Vm及Vd升高,PI及RI降低,H组HR及上述指标降低(P＜0.05);与S组相比,T1时H组HR及上述指标降低(P＜0.05).结论 0.5%利多卡因上胸段硬膜外交感神经阻滞可抑制交感神经兴奋,从而改善兔SAH后脑血管痉挛.     

细胞凋亡在蛛网膜下腔出血后脑血管痉挛发病中的作用

背景 脑血管痉挛(cerebral vasospasm,CVS)是蛛网膜下腔出血(subarachnoid hemorrhage,SAH)和颅内动脉瘤术后致死或致残的主要并发症之一,如何有效地治疗脑血管痉挛成为防治SAH的主要目标,但其确切机制尚不清楚,新近许多研究表明细胞凋亡可能在SAH后CVS的发病中发挥重要作用....     

不同时间浅低温对蛛网膜下腔出血犬脑血管痉挛的影响

目的 探讨不同时间浅低温对蛛网膜下腔出血犬脑血管痉挛的影响.方法 成年健康犬30只,雌雄不拘,体重14～20 kg,采用枕骨大孔二次注血法制备犬蛛网膜下腔出血模型.随机分为5组(n=6),人工脑脊液组(ACSF组):枕骨大孔处注入ACSF 0.5 ml/ks;蛛网膜下腔出血组(SAH组):制备蛛网膜下腔出血模型,维持直肠温度38～39℃;不同时间浅低温组(H_(1～3)组):在第2次注血后分别立即实施8、16和32 h浅低温(33.5℃),随后复温至38～39℃.于第1次注血或注入ACSF前和第2次注血或注入ACSF 5、12和19 d时行整体功能分级(OPC),并采用ELISA法和硝酸还原酶法分别测定血浆和脑脊液内皮素-1(ET-1)和一氧化氮(NO)代谢产物NO_2~-/NO_3~-的浓度,采用CT血管造影动态测量脑基底动脉直径.结果 与ACSF组比较,SAH组和H_1,2组OPC分级升高,血浆和脑脊液ET-1浓度升高,NO_2~-/NO_3~-浓度降低,基底动脉直径减小(P＜0.05),H_3组上述指标差异无统计学意义(P＞0.05);与SAH组比较,H_2,3组OPC分级降低,血浆和脑脊液ET-1浓度降低,NO_2~-/NO_3~-浓度升高,基底动脉直径增大(P＜0.05),H_1组各时点上述指标差异无统计学意义(P＞0.05);与H_2组比较,H_3组血浆和脑脊液ET-1浓度降低,NO_2~-/NO_3~-浓度升高,基底动脉直径增大(P＜0.05).结论 浅低温16～32 h可缓解蛛网膜下腔出血犬脑血管痉挛,且随浅低温时间延长该作用增强,其机制可能与降低全身和脑组织ET-1水平,升高NO水平,调节脑血管收缩-舒张平衡有关.     

基于MR血管成像比较两种兔蛛网膜下腔出血后脑血管痉挛模型

目的应用MRA对比观察一次注血法和二次注血法制造兔蛛网膜下腔出血后脑血管痉挛模型的差异。方法新西兰兔40只,随机分为对照组（5只）、一次注血组（17只）和二次注血组（18只）,分别以生理盐水（对照组）及自体动脉血（注血组）注入枕大池。于造模前和造模后1、3、5、7、9、11天行神经功能评分,采用MRA技术观测基底动脉管径的变化情况。结果 3组动物死亡率差异无统计学意义（均P〉0.05）。对照组无明显神经功能损伤,基底动脉管径造模前后差异无统计学意义（均P〉0.05）。一次注血组造模后3天神经功能损伤最明显,基底动脉于造模后1天发生收缩痉挛,于造模后3天痉挛达到高峰,管径为造模前的68.18%,后逐渐恢复正常。二次注血组造模后5天神经功能损伤症状最明显,基底动脉于造模后1天发生收缩痉挛,于造模后5天痉挛达到高峰,管径为造模前的56.72%,后逐渐恢复正常。结论兔枕大池二次注血法是制造蛛网膜下腔出血后脑血管痉挛模型的良好方法;采用MRA技术可活体观测脑血管痉挛。     

持续输注尼莫地平对颅内动脉瘤夹闭术中脑血管痉挛的影响

目的 了解颅内动脉瘤夹闭术中异氟醚麻醉复合输注尼莫地平对脑血管痉挛的影响。方法择期颅内动脉瘤夹闭术患者30例,随机表法分为两组:异氟醚组和尼莫地平组(各15例),异氟醚组术中吸入1个肺泡气最低有效浓度(MAC)异氟醚维持麻醉;尼莫地平组在诱导后输注尼莫地平20 μg·kg-1·h-1至手术结束后,同时吸入1 MAC异氟醚维持麻醉。术中于动脉瘤夹闭前、夹闭后即刻、2 h、4 h取脑脊液采用酶联免疫吸附试验法测定S100B含量;于动脉瘤夹闭前后测定载瘤动脉近心端及远心端血流速度。结果(1)尼莫地平组在动脉瘤夹闭前后脑脊液SIOOB含量无显著变化,而异氟醚组在动脉瘤夹闭后4 h S100B含量显著升高(F=4.11,P<0.05)。(2)尼莫地平组在动脉瘤夹闭前后载瘤动脉近心端血流速度[(15±9)与(19±8)cm／s]显著低于异氟醚组[(24±13)与(26±10)cm／s,t=2.08,P<0.05],而远心端血流速度无显著性差异。结论在颅内动脉瘤夹闭术中持续输注尼莫地平20μg·kg·-1·h-1对脑血管痉挛有一定预防作用。     

Brainstem hypoperfusion in severe symptomatic vasospasm following aneurysmal subarachnoid hemorrhage: role of basilar artery vasospasm

Summary Background. The hemodynamic effects of vertebrobasilar vasospasm are ill defined. The purpose of this study was to determine the effects of basilar artery (BA) vasospasm on brainstem (BS) perfusion. Methods. Forty-five patients with delayed ischemic neurological deficits (DIND) following aneurysmal subarachnoid hemorrhage (SAH) underwent cerebral angiography prior to decision-making concerning endovascular treatment. BA diameter was compared with baseline angiogram. Regional brainstem (BS) cerebral blood flow (CBF) was qualitatively estimated by ^99mTc ethyl cysteinate dimer single photon emission computed tomography (ECD-SPECT). Findings. Delayed BS hypoperfusion was found in 22 (48.9%) of 45 patients and BA narrowing of more than 20% was found in 23 (51.1%). Seventeen of 23 (73.9%) patients with BA narrowing of more than 20% experienced BS hypoperfusion compared to 6 of 22 (27.3%) patients with minimal or no narrowing (p = 0.0072). Patients with severe and moderate BS hypoperfusion had higher degree of BA narrowing compared to patients with normal BS perfusion and mild BS hypoperfusion (p < 0.001). The three-month outcome of patients n-22) with BS hypoperfusion was significantly worse compared to patients (n-23) with unimpaired (p = 0.0377, odd ratio for poor outcome 4, 1.15–13.9 95% confidence interval). Interpretation. These findings suggest that the incidence of BA vasospasm in patients with severe symptomatic vasospasm is high and patients with significant BA vasospasm are at higher risk to experience BS ischemia. Further studies should be done to evaluate the effects of endovascular therapy on BS perfusion and the impact of BS ischemia on morbidity and mortality of patients with severe symptomatic vasospasm.     

Review of aneurysmal subarachnoid hemorrhage—Focus on treatment,anesthesia, cerebral vasospasm prophylaxis,and therapy

Aneurysmal subarachnoid hemorrhage (aSAH) is a serious and debilitating condition that leads to the development of many complications, which are followed by mortality and morbidity. As anesthesiologists, we may require to manage aSAH at various settings such as in the perioperative period or in a nonoperative setting such as the neuroradiology suite for diagnostic and therapeutic interventions. Therefore, it is important to understand the pathophysiology of aSAH and anesthetic management for operations and interventions. For decades, early brain injury and cerebral vasospasm have played major roles in the outcome following aSAH. The purpose of this article is to review recent advances and future perspectives in the treatment of aSAH, early brain injury, and cerebral vasospasm.     

Effect of prostacyclin and adenosine triphosphate on vasospasm of canine basilar artery

Cerebral vasospasm is one of the most important factors influencing morbidity and mortality of intracranial operations or diseases. Platelet aggregation and adhesion is increased in spastic vessels. Degradation of platelets liberates mediators, which in turn increase vasospasm, thus creating a vicious cycle. Healthy vessels cope with this by increasing the synthesis of prostacyclin. The purpose of this study was to increase experimentally the levels of arterial prostacyclin and adenosine triphosphate (ATP) in animals through intraarterial injection of these substances because they are lower in spastic vessels. Prostacyclin promotes antiaggregation and dilatation, increases blood flow, inhibits thromboxane A2, and prevents synthesis of angiotensin II. Most of these effects were done by increasing cyclic adenosine monophosphate (cAMP). After injecting autogenous blood into the cisterna magna of male dogs, both the acute and chronic phases of vasospasm and the degenerative changes in the arterial wall were observed. Injecting ATP increased the severity of vasospasm. During vasospasm it was found that when prostacyclin is used intraarterially, vasodilatation began, but degeneration of the arterial wall could not be prevented. In the group of animals in which both ATP and prostacyclin were used, there was no degeneration of the arterial wall and the basilar artery was seen to be normal when viewed under the electron microscope.     

Effects of raloxifene on cerebral vasospasm after experimental subarachnoid hemorrhage in rabbits

Background

The aim of this study was to investigate the ability of a SERM, RLX, to prevent vasospasm in a rabbit model of SAH.

Methods

Thirty-four New Zealand white rabbits were allocated into 3 groups randomly. Subarachnoid hemorrhage was induced by injecting autologous blood into the cisterna magna. The treatment groups were as follows: (1) sham operated (no SAH [n = 12]), (2) SAH only (n = 12), and (3) SAH plus RLX (n = 10). Basilar artery lumen areas and arterial wall thickness were measured to assess vasospams in all groups.

Results

There was a statistically significant difference between the mean basilar artery cross-sectional areas and the mean arterial wall thickness measurements of the control and SAH-only groups (P < .05). The difference between the mean basilar artery cross-sectional areas and the mean arterial wall thickness measurements in the RLX-treated group was statistically significant (P < .05). The difference between the SAH group and the SAH + RLX group was also statistically significant (P < .05).

Conclusions

These findings demonstrate that RLX has marked vasodilatatory effect in an experimental model of SAH in rabbits. This observation may have clinical implications suggesting that this SERM drug could be used as possible anti-vasospastic agent in patients without major adverse effects.