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1.
目的探讨neuregulin基因产物胶质细胞生长因子(glial growth factor,GGF)在大鼠胚胎脊髓及正常成年脊髓内的表达及其在大鼠脊髓损伤后的表达变化及意义。方法采用改良Allen’S脊髓损伤打击模型,用逆转录一聚合酶链式反廊(RT—PCR)法检测胚胎大鼠脊髓内、正常成年大鼠脊髓内以及脊髓损伤后不同时间点伤段脊髓组织中GGFmRNA的表达。结果GGF在成年大鼠正常脊髓内有低水平表达,在胚胎脊髓中高水平表达。大鼠SCI后GGFmRNA的表达持续降低,在SCI后5d达到最低峰,之后GGFmRNA的表达可逐渐恢复。结论GGF在胚胎脊髓中高度表达,提示GGF在脊髓生长发育中起重要作用。大鼠脊髓损伤后GGF表达的降低可能与SCI后少突胶质细胞大量死亡或凋亡有关,与SCI后轴突无法再生、脱髓鞘的髓鞘再生困难有关。  相似文献   

2.
目的研究脂质体介导增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)和胶质细胞生长因子2(glial growth factor 2,GGF2)的双基因真核表达载体在脊髓内转染及其表达变化。方法构建GGF2与EGFP双基因真核表达载体pIRES2-EGFP-GGF2。采用注射法将阳离子脂质体和pIRES2-EGFP-GGF2质粒混合后转染至大鼠胸段脊髓组织中。采用RT-PCR法检测不同时间点转染区域脊髓组织中GGF2 mRNA的表达。在荧光显微镜下观察不同时间点EGFP在大鼠脊髓中的表达。结果基因转染后2 d即可观察到EGFP的表达及GGF2 mRNA表达的增加,在基因转染后1~2周内EGFP及GGF2 mRNA呈高水平表达,基因转染4周后,EGFP及GGF2 mRNA的表达明显减少。结论脂质体介导pIRES2-EGFP-GGF2转染大鼠脊髓后其表达的特点有助于基因治疗效果的即刻性,即早期大量表达,起到基因治疗的作用,又可避免外源基因的持续表达。  相似文献   

3.
目的 探讨细胞周期蛋白D3(Cyclin D3)在横断性脊髓损伤(tSCI)后的表达变化以及定位情况。方法 将48只成年SD大鼠随机分为8组:正常对照组,T9横断伤2、8h、1、3、5、7和14d组,每组6只。采用Western blot测定损伤后各时间段Cyclin D3蛋白水平在脊髓中的表达变化。采用免疫组织化学方法检测Cyclin D3在正常以及损伤后脊髓中的分布和定位。结果 West—emblot显示,Cyclin D3蛋白水平在tSCI后头、尾段均呈现先升高后下降的趋势,尾段明显:Cyclin D3的表达于损伤后8h开始逐渐升高,3d达到高峰,一直持续到第5天,之后逐渐下降。免疫组织化学表明Cyclin D3在正常脊髓中均匀分布,损伤后3d,Cyclin D3在脊髓白质和灰质中表达明显增强;免疫荧光双标记表明Cyclin D3与神经元的标记物neuronal nucleus(NeuN)、少突胶质细胞标记物cyclic nucleotide 3’phosphohydrolase(CNPase)有明显共定位,与星形胶质细胞标记物gtial fibrillary acidic protein(GFAP)和小胶质细胞标记物OX-42也存在部分共定位。结论脊髓损伤后Cyc—lin D3蛋白水平呈现明显的时相变化,并且与神经元、少突胶质细胞、星形胶质细胞和小胶质细胞存在共定位,提示Cyclin D3参与了脊髓损伤后的病理生理过程。  相似文献   

4.
目的 探讨大鼠脊髓损伤后脊髓神经干细胞的分离培养方法及分化情况.方法 采用Allen法制作大鼠脊髓损伤模型,利用无血清培养和单细胞克隆技术在成年脊髓损伤7 d大鼠脊髓中分离具有单细胞克隆能力的神经干细胞,并进行培养鉴定.结果 从成年脊髓损伤7 d大鼠脊髓中成功分离出神经干细胞,该细胞具有连续克隆能力,可传代培养,表达神经巢蛋白抗原.分化后的细胞表达神经元细胞、星形胶质细胞和少突胶质细胞的特异性抗原.结论 致伤7 d的成年大鼠脊髓组织体外町培养出神经十细胞,并分化为神经无细胞、星形胶质细胞和少突胶质细胞,有可能参与脊髓损伤的修复过程.  相似文献   

5.
胚胎脊髓细胞悬液植入急性成年大鼠损伤脊髓   总被引:6,自引:2,他引:4  
目的:建立胚胎脊髓细胞悬液移植于脊髓损伤模型,以评价其治疗脊髓损伤的可能性。方法:42只Wistar大鼠以改良Alen法(50gcm)打击脊髓,3天后将孕14天(E14)FSCS20μl植入损伤空腔,移植后2、4、6、8、10、12周,以光、电镜、免疫组织化学观察移植物存活、分化及其与宿主之间关系。结果:移植细胞逐渐长大。充满不规则空腔,宿主NF、5-HT、CGRP纤维分别出现于移植物,GFAP纤维于宿主移植物交界处适量存在。移植成神经细胞、成少突胶质细胞、成星形细胞的细胞器日渐完善,细胞功能活跃。复杂及多样突触与细胞连接,将上述细胞与神经纤维、胶质纤维、毛细血管网在三维空间内连接成一体,并与宿主紧密嵌合。结论:(1)成年大鼠脊髓损伤3天后植入FSCS可以存活。(2)移植物进入宿主后,出现再分布,继而器官样分化。(3)长、短传导束进入移植物,显示了移植物的桥作用。(4)成少突胶质细胞的神经营救作用。(5)移植区内出现多种突触,提示移植物中继作用的可能性。  相似文献   

6.
脊髓损伤后促红细胞生成素对bcl-2的影响   总被引:1,自引:0,他引:1  
目的 探讨促红细胞生成素(EPO)对大鼠脊髓损伤后伤区脊髓细胞凋亡和神经功能恢复的影响。方法 Wistar大鼠210只,随机分为4组:假手术组、脊髓损伤组、脊髓损伤加重组人EPO治疗组、脊髓损伤加生理盐水治疗组。采用原位末端脱氧核糖核苷酸转移酶介导dUTP标记(TUNEL标记法)检测神经元和少突胶质细胞凋亡,Western blot免疫印迹法和免疫组化染色检测bcl-2表达,免疫组化染色和图像分析方法观察对白质内神经纤维(NF-200染色)的保护作用,通过感觉诱发电位(SSEP)、运动诱发电位(MEP)和大鼠BBB后肢运动功能评分,观察损伤脊髓传导功能的恢复。结果 EPO保护组bcl-2在各时相点的表达明显增高,8h和7d时神经元和少突胶质细胞的TUNEL阳性细胞数明显减少;在7d时白质中NF-200阳性神经纤维明显增多;SSEP和MEP的平均潜伏期和波幅以及BBB功能评分明显提高,与损伤组和生理盐水治疗相比,差异有统计学意义(P〈0.01)。结论 EPO通过上调bcl-2的表达,在抑制脊髓损伤后神经元和少突胶质细胞的凋亡中起到神经保护作用。  相似文献   

7.
[目的] 研究大鼠胚胎脊髓移植后是否能够影响生长抑素(SOM)mRNA的表达。[方法]将动物分为单纯半切洞损伤组(A组)、胚胎脊髓移植组(B组)。术后1、3、7、14、28d,采用原位杂交方法观察SOM mRNA的表达,采用计算机图像分析技术,进行定量分析。[结果] 胚胎脊髓移植组SOM mRNA蛋白表达明显多于单损伤组。[结论] 作者的研究表明胚胎脊髓移植后可使损伤脊髓SOM mRNA表达增多。  相似文献   

8.
体外转基因成肌细胞移植对大鼠损伤脊髓细胞凋亡的影响   总被引:1,自引:1,他引:1  
目的:探讨大鼠脊髓损伤后胚胎脊髓和腺病毒介导的脑源性神经生长因子(AxCA-BDNF)体外转基因成肌细胞移植对大鼠脊髓细胞凋亡的影响。方法:将动物分为:大鼠脊髓半切洞损伤明胶海绵填充组(A组),大鼠脊髓半切洞损伤应用胚胎脊髓移植组(B组),脊髓半切洞损伤损伤AxCA-BDNF基因转染的成肌细胞移植组(C组)大鼠脊髓半切洞损伤后应用胚胎脊髓和AxCA-BDNF基因转染的成肌细胞移植组(D组)。手术后1、3、7、14、28d应用行为学和电生理检查观察大鼠功能恢复情况,对脊髓损伤区进行细胞凋亡的检测(TUNEL)以及Bcl-2蛋白表达的测定(免疫组化法)。采用计算机图像分析技术,进行定量分析。结果:A、B、C、D四组中均发现凋亡细胞及Bcl-2蛋白阳性表达细胞,图像分析发现,各组凋亡细胞核为A>B>C>D;Bcl-2免疫反应阳性细胞表达顺序为D>C>B>A,Bcl-2免疫反应阳性细胞的表达与大鼠后肢功能恢复有同样的变化趋势。结论:大鼠胚胎脊髓和体外转基因成肌细胞移植能抑制脊髓损伤后的细胞凋亡。  相似文献   

9.
目的观察大鼠胚胎脊髓移植对损伤脊髓 GAP- 43 mRNA表达和大鼠后肢功能恢复的影响。方法成年 Wistar大鼠 66只,随机被分为脊髓半切洞损伤应用胚胎脊髓移植组 (脊髓移植组, 30只 )、单纯脊髓半切洞损伤明胶海绵填充组 (单纯损伤组, 30只 )、正常对照组 (6只 )。术后第 1、 3、 7、 14、 28 d,对大鼠进行 Tarlov评分和斜板试验检查后肢功能。应用原位杂交的方法观察 GAP- 43 mRNA的表达,记录阳性细胞数,采用计算机图像分析技术,进行定量分析。结果脊髓损伤后第 1 d,在脊髓移植组和单纯损伤组的损伤脊髓灰质中都可见到 GAP- 43 mRNA的表达。于损伤后迅速增加,第 7 d时达到高峰。脊髓移植组 GAP- 43 mRNA蛋白表达为 39.24± 6.83,约是单纯损伤组 (21.48± 7.83)的 2倍 (P< 0.05)。胚胎脊髓移植后可使损伤的脊髓中 GAP- 43 mRNA持续高表达至术后第 28 d,而单纯损伤组仅持续到术后第 14 d。增加的 GAP- 43 mRNA阳性细胞数与神经功能的改善平行。结论胚胎脊髓移植后可使损伤大鼠脊髓高表达 GAP- 43 mRNA,并促进大鼠功能的恢复。  相似文献   

10.
神经生长导向因子Slit2在成年大鼠急性脊髓损伤后的表达   总被引:2,自引:0,他引:2  
目的观察Slit2在大鼠急性脊髓损伤后不同时点表达量的变化和分布,探讨轴突再生导向机制。方法成年SD大鼠采用Allen’s法制作脊髓损伤(SCI)模型,分别于损伤后第2、4、7、14天取材,半定量逆转录-聚合酶链反应(RT-PCR)分析Sht2 mRNA表达水平变化,免疫组织化学法观察Slit蛋白在损伤脊髓区的分布情况。结果RT—PCR提示脊髓损伤后Slit2即开始出现转录上调,表达量持续增高并于第7天达到顶峰,之后逐步下降;免疫组织化学提示Slit2阳性信号先后出现在急性SCI区灰质胶质细胞及神经元胞质中,以前、后角及中央管周围较为聚集。结论大鼠SCI后有Slit2表达上调出现并在损伤脊髓灰质广泛分布,可能是轴索再生过程中生长锥导向性生长的关键调节者。  相似文献   

11.
Background : We investigated the vasopressor hormone response following mesenteric traction (MT) with hypotension due to prostacyclin (PGI2) release in patients undergoing abdominal surgery with a combined general and epidural anesthesia. Methods : In a prospective, randomized, placebo-controlled study we administered 400 mg ibuprofen (i.v.) in 42 patients scheduled for abdominal surgery. General anesthesia was combined with epidural anesthesia (T4-L1). Before as well as 5, 15, 30, 45, and 90 min after MT we recorded plasma osmolality, hemodynamics and measured 6-keto-PGFlα (stabile metabolite of PGI2), TXB2 (stabile metabolite of thromboxane A2) active renin, and arginine vasopressin (AVP) plasma concentrations by radioimmunoassay. Catecholamine levels were assessed by high-pressure liquid chromatography (HPLC) with electrochemical detection. Results : Following MT, arterial hypotension occurred along with a substantial PGI2 release. This was completely abolished by ibuprofen administration. Although plasma levels of 6-keto-PGF (1133 (708) vs. 60 (3) ng/L, median (median absolute deviation), P=0.0001, placebo vs. ibuprofen) remained significantly elevated, blood pressure was restored within 30 min after MT in the placebo group. At the same point in time plasma concentrations of TXB2 (164 (87) vs. 58 (1) ng/L, P=0.0001), epinephrine (46 (33) vs. 14 (6) ng/L, P=0.001), AVP (41 ± (18) vs. 12 (7) ng/L, P=0.0004), and active renin (27 (12) vs. 12 (4) ng/L, P = 0.001) were significantly higher in placebo-treated patients. Conclusion : Under combined general and epidural anesthesia arterial hypotension following MT due to endogenous PGI2 release is associated with enhanced release of AVP, active renin, epinephrine and thromboxane A2, presumably contributing to hemodynamic stability within 30 min after MT.  相似文献   

12.
Don Dame 《Artificial organs》1996,20(5):613-617
Abstract: Virtually all blood pumps contain some kind of rubbing, sliding, closely moving machinery surfaces that are exposed to the blood being pumped. These valves, internal bearings, magnetic bearing position sensors, and shaft seals cause most of the problems with blood pumps. The original teaspoon pump design prevented the rubbing, sliding machinery surfaces from contacting the blood. However, the hydraulic efficiency was low because the blood was able to "slip around" the rotating impeller so that the blood itself never rotated fast enough to develop adequate pressure. An improved teaspoon blood pump has been designed and tested and has shown acceptable hydraulic performance and low hemolysis potential. The new pump uses a nonrotating "swinging" hose as the pump impeller. The fluid enters the pump through the center of the swinging hose; therefore, there can be no fluid slip between the revolving blood and the revolving impeller. The new pump uses an impeller that is comparable to a flexible garden hose. If the free end of the hose were swung around in a circle like half of a jump rope, the fluid inside the hose would rotate and develop pressure even though the hose impeller itself did not "rotate"; therefore, no rotating shaft seal or internal bearings are required.  相似文献   

13.
Abstract: A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal.  相似文献   

14.
Background : Our objective was to determine whether administration of propranolol or verapamil modifies the hemodynamic adaptation to continuous positive-pressure ventilation (CPPV), in particular the regional distribution of cardiac output (CO).
Methods : General hemodynamics and regional blood flows assessed by microsphere technique (15 (μm) were recorded in 16 anesthetized pigs during spontaneous breathing (SB) and CPPV with 8 cm H2O end-expiratory pressure (CPPV8) before and after intravenous administration of propranolol (0.3 mg · kg−1 followed by 0.15 mg · kg−1 · h−1, n=8) or verapamil (0.1 mg · kg−1 followed by 0.3 mg · kg−1 · h−1, n=8).
Results : CPPV8 depressed CO by 25% without shifts in its relative distribution with the exception of a noteworthy increase in adrenal perfusion. Propranolol increased arterial blood pressure, and due to a fall in heart rate, CO dropped by 25%. The kidneys and, to a lesser extent, the splanchic region and central nervous system received increased fractions of the remaining CO at the expense of skeletal muscle flow. Similar patterns were seen during SB and CPPV8 such that the combination of propranolol and CPPV8 depressed CO by 50%. The circulatory effects of verapamil were less evident but myocardial perfusion tended to increase.
Conclusions : The combination of propranolol or verapamil with CPPV does not result in any specific hemodynamic interaction in anesthetized pigs, except that the combined effect of propranolol and CPPV may severely reduce CO.  相似文献   

15.
Background: Obesity is increasing globallly, including in the formerly "Eastern Bloc" countries. Methods: A survey was made of obesity and bariatric surgery. Results: In the 8 East and Central European countries studied, with total population 300 million, roughly 43% of the population was overweight (BMI 25-30), 23% obese (BMI > 30), with about 15 million people morbidly obese (BMI > 40). From 0-10 morbidly obese individuals/100,000/year undergo bariatric surgery. Conclusion: Most countries were found to provide inadequate treatment for obesity.The majority of the morbidly obese are not treated effectively. However, health-care awareness of obesity and bariatric surgeons are slowly increasing.  相似文献   

16.
Background : Inhibitory effects of volatile anaesthetics on platelet aggregation have been demonstrated in several studies. However, the influence of volatile anaesthetics on intracoronary platelet adhesion has not been elucidated so far.
Methods : Isolated hearts of guinea pigs were perfused with buffer in the absence or presence of volatile anaesthetics (0.5 and 1 MAC) at constant coronary flow rates of 5 ml/min for 25 min, then 1 ml/min for 30 min and again 5 ml/min for 10 min. Before, during and after low-flow perfusion, a bolus of human platelets was applied into the coronary system. To simulate thrombogenic conditions, 0.3 U/ml human thrombin was infused during low-flow perfusion and reperfusion. The number of platelets sequestered to the endothelium was calculated from the difference between coronary in- and output of platelets. The myocardial production of lactate and consumption of pyruvate and coronary perfusion pressure were also determined.
Results : At a flow rate of 5 ml/min only about 3% of the applied platelets did not emerge from the coronary system, in any group. In contrast, 13.1±1.2% (mean±SEM) of infused platelets became adherent in low-flow perfusion in the control group without anaesthetic. The adherence was reduced with each 1 MAC isoflurane (to 6.2±1.2%), sevoflurane (to 4.4±0.9%) or halothane (to 3.2±1.5%) (each P <0.05 vs. control). Volatile anaesthetic, 0.5 MAC, did not inhibit platelet adhesion to a statistically significant extent in any case. Perfusion pressure and metabolic parameters were not statistically different between the control and the hearts exposed to anaesthetics.
Conclusion : Volatile anaesthetics in a concentration of 1 MAC can reduce the adhesion of platelets in the coronary system under reduced flow conditions. This action does not arise from vasodilation or inhibition of ischaemic stress.  相似文献   

17.
Background: It has been shown that the depressive effects of both propofol and midazolam on consciousness are synergistic with opioids, but the nature of their interactions on other physiological systems, e. g. respiration, has not been fully investigated. The present study examined the effect of propofol and midazolam alone and in combination with fentanyl on phrenic nerve activity (PNA) and whether such interactions are additive or synergistic. Methods: PNA was recorded in 27 anaesthetised and artificially ventilated rabbits. In three groups, propofol, fentanyl and midazolam were administered intravenously in incremental doses to construct dose-response curves for the depressant effects of each one on PNA. In another two groups, the effect of pretreatment with either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. on the effects of propofol and fentanyl respectively on PNA were studied. Results: Propofol and fentanyl caused a dose-dependent depression of PNA with complete abolition at the highest total doses of 16 mg · kg?1 i. v. and 32 μg · kg?1 i. v., respectively. In contrast, midazolam in incremental doses to a total of 0.8 mg · kg?1 reduced mean PNA by 63%, but approximately 12% of PNA remained at a total dose as high as 6.4 mg · kg?1. The mean ED50s, calculated from dose-response curves, were 5.4 mg · kg?1, 3.9 μg · kg?1 and 0.4 mg · kg?1 for propofol, fentanyl and midazolam, respectively. Initial doses of either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. acted synergistically with subsequent doses of either propofol or fentanyl to abolish PNA at total doses of 8 mg · kg?1 and 8 μg · kg?1, respectively. Conclusion: Fentanyl has a synergistic interaction with both propofol and midazolam on PNA and hence potentially on respiration.  相似文献   

18.
Background: Catecholaminergic support is often used to improve haemodynamics in patients undergoing major abdominal surgery. Dopexamine is a synthetic vasoactive catecholamine with beneficial microcirculatory properties. Methods: The influence of perioperative administration of dopexamine on cardiorespiratory data and important regulators of macro- and microcirculation were studied in 30 patients undergoing Whipple pancreaticduodenectomy. The patients received randomized and blinded either 2 μg · kg?1 · min?1 of dopexamine (n=15) or placebo (n=15, control group). The infusion was started after induction of anaesthesia and continued until the morning of the first postoperative day. Endothelin-1 (ET-1), vasopressin, atrial natriuretic peptide (ANP), and catecholamine plasma levels were measured from arterial blood samples. Measurements were carried out after induction of anaesthesia, 2 h after onset of surgery, at the end of surgery, 2 h after surgery, and on the morning of the first postoperative day. Results: Cardiac index (CI) increased significantly in the dopexamine group (from 2.61±0.41 to 4.57±0.78 1 · min?1 · m?2) and remained elevated until the morning of the first postoperative day. Oxygen delivery index (DO2I) and oxygen consumption index (VO2I) were also significantly increased in the dopexamine group (DO2I: from 416±91 to 717±110 ml/m2 · m2; VO2I: from 98±25 to 157±22 ml/m2 · m2), being significantly higher than in the control group. pHi remained stable only in the dopexamine patients, indicating adequate splanchnic perfusion. Vasopressive regulators of circulation increased significantly only in the untreated control patients (vasopressin: from 4.37±1.1 to 35.9±12.1 pg/ml; ET-1: from 2.88±0.91 to 6.91±1.20 pg/ml). Conclusion: Patients undergoing major abdominal surgery may profit from prophylactic perioperative administration of dopexamine hydrochloride in the form of improved haemodynamics and oxygenation as well as beneficial influence on important regulators of organ blood flow.  相似文献   

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A concept of balanced analgesia using nonsteroidal anti-inflammatory drugs (NSAIDs), paracetamol (acetaminophen), opioids, and corticosteroids can also be used in patients with pre-existing illnesses. NSAIDs are the most effective treatment for acute pain of moderate intensity in children; however, these drugs should be avoided in patients at increased risk for serious side effects, e.g. patients with renal impairment, bleeding tendency, or extreme prematurity. NSAIDs can be given with minimal risks to the younger child with mild to moderate asthma, and, in these patients, the use of steroids can be encouraged; in addition to their antiemetic and analgesic action, a beneficial effect on asthma symptoms can be expected. In the non-intubated child with cerebral trauma, exaggerated sedation caused by opioids and increased bleeding tendency caused by NSAIDs must be avoided. In neonates and small infants, the oral administration of sucrose or glucose is helpful to minimize pain reaction during short uncomfortable interventions.  相似文献   

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