脑缺血/再灌注损伤对小鼠海马区胶质细胞的影响

目的探讨小鼠脑缺血/再灌注后胶质细胞形态学与数量的变化及其临床意义。方法采用随机方法将32只健康昆明雄性小鼠分为假手术组和模型组,每组16只。制备脑缺血/再灌注损伤模型,应用免疫组织化学方法分别标记星形胶质细胞特异性蛋白胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)与小胶质细胞特异性蛋白离子钙接头蛋白抗体(ionized calcium-binding adaptor molecule 1,Iba-1),观察脑缺血/再灌注小鼠海马区胶质细胞形态及数量的变化。结果脑缺血/再灌注损伤后模型组小鼠海马区锥体细胞排列紊乱,细胞脱失明显,细胞核体积变小,深染,呈核固缩。GFAP与Iba-1阳性细胞在形态上表现为胞体肥大,分支变粗;模型组阳性细胞数分别为(12.56±1.58)个/HP和(9.63±1.50)个/HP,均显著多于假手术组,差异具有统计学意义(P<0.01)。结论小鼠局灶性脑缺血/再灌注损伤后海马区胶质细胞过度活化,可能参与加重脑缺血/再灌注后组织损伤及神经元凋亡。     

抗呆1号对脑缺血再灌注小鼠海马星形胶质细胞GFAP表达的影响

应用免疫组织化学方法探讨抗呆Ⅰ号对小鼠前脑缺血再灌注损伤后海马星形胶质细胞表达GFAP动态变化的影响,其与缺血性神经元的联系.结果显示:前脑缺血再灌注1 d只有少数星形胶质细胞表达GFAP,再灌注3 d表达GFAP的阳性细胞数增加,再灌注7～10 d表达GFAP的阳性细胞数达到高峰,同时,可见细胞反应性胶质化特征,前脑缺血再灌注15 d GFAP阳性反应细胞数仍维持在较高水平.用药组GFAP的表达细胞数较模型组明显减少.而正常对照组和假手术组只有少数星形胶质细胞表达GFAP.且星形胶质细胞表达GFAP的强弱与神经细胞的存亡有关.说明前脑缺血再灌注后海马反应性星形胶质细胞表达GFAP呈动态变化,抗呆Ⅰ号对缺血再灌注损伤星形胶质细胞和神经细胞有良好的保护作用.     

黄精口服液对血管性痴呆大鼠行为和海马星形胶质细胞的影响

目的 观察大鼠学习记忆能力和海马星形胶质细胞的变化,探讨黄精口服液对血管性痴呆大鼠的干预效果.方法 应用Morris水迷宫、免疫组织化学法和透射电镜等技术观察血管性痴呆大鼠海马组织胶质纤维酸性蛋白(GFAP)的表达和CA1区星形胶质细胞超微结构的变化.结果 (1)术后3.5个月,痴呆+黄精口服液组大鼠平均逃避潜伏期[(21.5±6.6)s]较血管性痴呆组[(31.8±7.2)s]和痴呆+盐水组[(31.0±9.2)S]减少(P＜0.01).(2)血管性痴呆组和痴呆+生理盐水组大鼠海马结构GFAP免疫反应阳性细胞数增多、校正灰度值比空白对照组增加(P＜0.01);痴呆+黄精口服液组大鼠海马结构GFAP免疫反应阳性细胞数及其胞浆校正灰度值[CA1辐射层、腔隙层和齿状回分子层分别为(27.3±7.6),(28.1±5.8),(35.4±9.6)]较痴呆+生理盐水组大鼠[(53.6±8.1),(57.9±6.4)和(59.7±6.5)]减少(P＜0.01).(3)血管性痴呆组和痴呆+生理盐水组大鼠海马组织微血管周围的星形胶质细胞肿胀,细胞内线粒体变形、肿胀、嵴断裂,胞质内大量溶酶体形成;并见空泡化的胶质细胞足突和退变的星形胶质细胞胞体.痴呆+中药组大鼠海马CA.区中,毛细血管周围的星形胶质细胞突起有轻度水肿,局部区域胶质细胞增生.结论 黄精口服液可抑制海马CA.区星形胶质细胞反应,减少其终足水肿,改善学习记忆能力.     

七氟烷后处理对小鼠脑缺血/再灌注损伤的作用及影响机制研究

目的 探讨七氟烷后处理对小鼠脑缺血/再灌注损伤的作用和影响机制.方法 选取首都医科大学动物中心健康雄性BALB/c小鼠,随机数字表法分为5组,每组15只,假手术组(Sham组)、缺血/再灌注组(ischemia/reperfusion,I/R组)、I/R+七氟烷组(I/R+ Sev组)、I/R+鞘氨醇激酶-2(sphingosine kinase 2,Sphk2)选择性抑制剂ABC294640组(I/R+ ABC组)和I/R+七氟烷+Sphk2选择性抑制剂ABC294640组(I/R+ Sev+ ABC组).采用大脑中动脉阻塞法(middle cerebral artery occlusion,MCAO)造成脑缺血损伤,缺血60 min后恢复脑灌注24h,建立I/R模型.Sev(3％,2 L/min)均于再灌注开始即刻给予,持续吸入1 h;ABC294640均于脑缺血模型制备前从侧脑室注入,剂量为8μl(15 nmol).缺血再灌注24h,进行神经行为学评分.随后处死小鼠,各组取5只小鼠分别测脑含水量、缺血半影区细胞凋亡数,并采用Western blot方法检测缺血半影区Sphk2、活化caspase-3和环氧合酶-2 (cyclooxygenase-2,COX2)表达水平.结果 与Sham组比较,I/R组神经功能缺陷评分[(0.0±0.0)分比(3.9±0.5)分]、脑含水量[(68.5±7.3)％比(86.2±9.1)％]、细胞凋亡数[(4±1)个/HP比(50±5)个/HP]、活化caspase-3 [(36.1±4.2)％比(81.2±9.2)％]和COX2 [(39.1±5.2)％比(95.3±10.1)％]均升高(P＜0.05),Sphk2 [(25.1±3.1)％比(19.2±3.1)％]表达降低(P＜0.05);与I/R组比较,I/R+ Sev组神经行为学评分[(3.9±0.5)比(2.4±0.3)]、脑含水量[(86.2±9.1)％比(75.2±8.1)％]、细胞凋亡数[(50±5)个/HP比(19±3)个/HP]、活化caspase-3 [(81.2±9.2)％比(50.3±6.1)％]和COX2 [(95.3±10.1)％比(52.2±6.4)％]表达均降低(P＜0.05),Sphk2 [(19.2±3.1)％比(49.3±5.2)％]表达升高(P＜0.05);与I/R+ Sev组比较,I/R+ Sev+ ABC组神经行为学评分[(2.4±0.3)比(3.3±0.4)]、脑含水量[(75.2±8.1)％比(82.3±9.1)％]和细胞凋亡数[(19±3)个/HP比(30±4)个/HP]、活化caspase-3 [(50.3±6.1)％比(63.2±7.3)％]和COX2 [(52.2±6.4)％比(78.4±8.3)％]表达均增高(P＜0.05),Sphk2[(49.3±5.2)％比(30.2±4.2)％]表达降低(P＜0.05).结论 七氟烷后处理可抑制细胞凋亡而减轻脑损伤,机制可能与Sphk2激活有关.     

补阳还五汤对脑缺血再灌注后星形胶质细胞connexin43影响

目的:探讨补阳还五汤对大鼠脑缺血再灌注后海马CA1区缝隙连接小体connexin43(Cx43)的影响。方法:采用颈内动脉线栓法建立大脑中动脉栓塞模型,脑缺血2 h再灌注1、3和7 d后,免疫组织荧光染色观察海马CA1区Cx43的表达情况,并通过星形胶质细胞标记物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的免疫染色比较星形胶质细胞的激活程度。结果:补阳还五汤组脑缺血再灌注1 d后海马CA1区Cx43的表达较模型组显著降低(P<0.05),而第3天和第7天海马CA1区Cx43的表达较模型组高(P<0.05)。同时,补阳还五汤组缺血再灌注1 d后星形胶质细胞的激活较模型组受到抑制;第3天和第7天星形胶质细胞激活均较第1天明显减弱,而且第7天时两组无明显差异。结论:补阳还五汤可能通过调控星形胶质细胞Cx43表达对脑缺血再灌注后不同时间点的损伤和修复进行调控。     

环磷酰胺对脑缺血再灌注海马GFAP表达的影响

目的观察环磷酰胺对脑缺血再灌注后大鼠大脑海马区胶原纤维酸性蛋白（GFAP）表达的影响。方法设正常组、假手术组、缺血再灌注组（I／R）和环磷酰胺处理组（T）,以改良Longa线栓法建立sD大鼠局灶性脑缺血模型。处理组缺血后1h给予环磷酰胺腹腔注射,处理组和I／R组均于缺血后2h形成再灌注,每组均分为三个时间段（24h、72h和5d）分别观察。应用免疫组织化学技术检测各组脑组织不同时段以及正常脑组织中海马GFAP的表达情况。结果脑缺血再灌注5d内,随着缺血时间的延长,GFAP在海马区星形胶质细胞中的阳性表达逐渐增多,缺血24h后即有表达,缺血再灌注72h后增多,5d后表达最高;经过环磷酰胺处理后,海马区GFAP表达减少（P〈0．05）。正常组及假手术组未见GFAP表达。结论大鼠脑缺血再灌注后5d内GFAP在海马中的表达与缺血时间呈明显的相关性,随着缺血时间延长而增加。用环磷酰胺处理后GFAP表达明显减少,提示环磷酰胺对缺血诱导的星形胶质细胞活化具有抑制作用,可能对脑缺血再灌注损伤的恢复起重要作用。     

间歇性低压缺氧预处理对大鼠全脑缺血/再注后海马神经元的保护作用

目的 探讨间歇性低压缺氧预处理对大鼠全脑缺血/再注后海马神经元的保护作用.方法 72只健康雄性Wistar大鼠,体重280～300g,随机分为6组,每组12只.分别为假手术组(Sham);间歇性低压缺氧预处理组(IHHP);缺血/再灌注组(I/R);间歇性低压缺氧预处理2次+全脑缺.血/再灌注组( IHHP2+ I/R);间歇性低压缺氧预处理4次+全脑缺血/再灌注组(IHHP4+I/R);间歇性低压缺氧预处理6次+全脑缺血/再灌注组(IHHP6+ I/R).采用四血管闭塞法复制大鼠全脑缺血/再灌注模型,全脑缺血后8 min行再灌注.硫堇染色观察海马CA1区组织学分级及锥体神经元密度;流式细胞技术检测海马神经元凋亡率.结果 与Sham组相比,单纯性间歇性低压缺氧预处理对海马组织形态和神经元凋亡率均无明显影响;I/R组大鼠海马CA1区组织损伤明显,存活的锥体细胞稀疏,排列紊乱,细胞明显缺失.I/R组与Sham组相比,组织学分级明显升高,神经元凋亡率明显增加;间歇性低压缺氧预处理2次、4次和6次+ I/R各组大鼠海马CA1区细胞损伤均明显改善,组织学分级明显降低,存活神经元密度值均明显增加,神经元凋亡率明显降低,其中IHHP4+ I/R组效果最为明显.结论 间歇性低压缺氧本身对大鼠海马神经元无明显损伤作用,但可对其后短期内发生的缺血/再灌注脑组织发挥保护作用,适当的预处理是诱导有效脑保护作用发生的必要条件.     

全脑缺血再灌注损伤大鼠海马区Skp2的表达变化

目的观察大鼠全脑缺血再灌注损伤后海马区S期激酶相关蛋白2(S phase kinase-associated protein 2,Skp2)及其下游底物p27的表达变化。方法雄性Wistar大鼠60只随机分为2组:对照组(n=20),全脑缺血再灌注组(n=40)。采用Pulsinelli四血管结扎法建立大鼠全脑缺血再灌注损伤模型,全脑缺血30min后进行再灌注;对照组为假手术组,只分离血管。分别于大鼠全脑缺血再灌注4、7d用免疫组织化学法检测海马区星形胶质细胞和神经元变化;Western blot方法检测海马区Skp2及p27蛋白表达变化。结果与对照组相比,大鼠全脑缺血再灌注4、7d后,星形胶质细胞明显活化、增殖;海马CA1区神经元在再灌注7d时明显减少。Western blot检测结果显示,与对照组相比,在全脑缺血再灌注4d和7d海马区Skp2表达逐渐增高,而p27蛋白的表达逐渐降低(均P<0.05)。结论全脑缺血再灌注损伤大鼠海马区Skp2的表达升高,可能与缺血后神经元凋亡及胶质细胞活化有关。     

瘦素对小鼠局灶性脑缺血再灌注损伤的保护作用及表达Cx43的影响

目的探讨瘦素对局灶性脑缺血再灌注损伤大脑缝隙连接蛋白43(Cx43)表达的影响及瘦素的脑保护作用机制。方法将45只健康雄性昆明小鼠随机分为假手术组、模型组和瘦素组。模型组和瘦素组制备小鼠大脑中动脉局灶性脑栓塞模型,瘦素组于缺血0min时腹腔注射1mg/kg瘦素,缺血2h再灌注24h后评价神经功能状态,脑组织TTC染色测定脑梗死体积,HE染色观察组织病理学改变;原代培养SD乳鼠的大脑皮层星形胶质细胞(Ast),将纯化后的Ast分为对照组、模型组、瘦素100μg/L组和瘦素500μg/L组,制作大鼠大脑皮质星形胶质细胞缺氧复氧模型,MTT法检测星形胶质细胞存活率,Western blot方法检测脑组织及星形胶质细胞中Cx43的表达变化。结果与模型组相比,瘦素组神经功能损伤状态得到改善(P<0.05),脑缺血再灌注后脑梗死体积缩小(P<0.05),脑组织病理学损伤程度减轻,Cx43的表达明显降低(P<0.01);与模型组比较,瘦素100μg/L组Ast存活率提高,Cx43表达减少,但差异均无统计学意义(P>0.05),瘦素500μg/L组Ast存活率显著提高(P<0.01),Cx43表达明显减少(P<0.01)。结论体内和体外实验均提示,瘦素可能通过降低损伤后脑组织中Cx43的表达,减轻脑缺血再灌注损伤。     

与缝隙连接43蛋白相关的星形胶质细胞通过外泌体对神经元保护作用的机制研究

目的:观察缝隙连接43(Cx43)蛋白参与星形胶质细胞来源的外泌体对神经元保护作用的机制.方法:采用C57BL/6胎鼠以原代培养法获取星形胶质细胞及神经元,建立共培养(Co-Culture)模型,给予神经元过氧化氢(H2O2)损伤,星形胶质细胞给予Cx43特异性阻断剂Gap26,分别建立(Co-Culture+H2O2)组及(Co-Culture+H2O2+Gap26)组,同时设单纯神经元损伤(Neuron+H2O2)组,通过蛋白免疫印记(WB)法,测定神经元骨架相关的紧密连接蛋白(ZO-1)、α-肌动蛋白(α-actin)、谷氨酸转运蛋白-1(GLT-1)、外泌体标记蛋白CD63和凋亡相关的Bax/Bcl-2比例的变化,采用高效液相色谱仪(HPLC)观察神经元谷氨酸(Glu),采用ELFA法检测氧化应激因子腺嘌呤二核苷酸磷酸(NAPDH)氧化酶活性.结果:与(Neuron+H2O2)组相比,(Co-Culture+H2O2)组的神经元ZO-1、α-actin、GLT-1和CD63的表达明显上调,而Bax/Bcl-2比例、NAPDH氧化酶活性则明显降低(P<0.05).在给予Gap26后,可明显抑制以上星形胶质细胞对神经元的此种作用(P<0.05).结论:Cx43蛋白可促进星形胶质细胞的外泌体被神经元吸收,可产生稳定细胞形态结构、加强兴奋性氨基酸转运及降低神经元凋亡和氧化应激损伤等神经保护功能.     

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.     

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…     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.