三线定位法在大鼠心脏采血中的应用

目的介绍一种改良的大鼠心脏穿刺采血方法。方法采用90只SPF级Wistar大鼠,雌雄各半,以10%水合氯醛溶液,按0.3 mL/kg体质量行腹腔注射麻醉,三线定位确定穿刺位点,使用采血针采血,利用采血管负压收集血液,记录每只大鼠采血量。结果以单只大鼠采血量达5 mL或以上为终末次采血成功计算成功率,90只大鼠共计采血90次,成功率约为82.2%,离心后血清总量约为260 mL,平均每只大鼠分离血清约为2.8 mL。结论改进后的心脏采血技术定位方法直观易学,且采血量多、血液质量高,同时创伤小、效率高,值得向广大同行推广应用。     

自制大鼠固定装置在大鼠断尾采血中的应用

目的探讨应用简易自制新装置进行大鼠断尾采血的效果,建立一种安全、实用、可重复的大鼠采血方法。方法用塑料瓶、木板等制作简易方便的装置固定大鼠,对鼠尾清洗、水浴等合适处理后进行断尾采血。结果自制固定器制作简单、操作方便,1 d可对20只大鼠重复采血6次,出血量0.5~1.0 m L,无大鼠死亡及操作人员受伤等情况发生。结论应用自制大鼠固定装置大鼠采血,血量多、安全性好,是一种理想的大鼠断尾采血新装置。     

SeV18+hFGF2/dF反复给予大鼠的毒性研究

目的:考察SeV18 hFGF2/dF(碱性成纤维细胞生长因子仙台病毒载体)的安全性。方法:本研究采用W istar大鼠,120只,雌性各半,随机分为4组分别给予剂量为1×108、4×108及4×109C IU.kg-1的SeV18 hFGF2/dF和磷酸盐缓冲液。肌肉注射给药,每天给药1次,连续给药14天,停药后继续观察4周。本报告总结了给药后对大鼠生理、生化、脏器病理指标的影响,检测了给药后动物抗体的产生情况。结果:反复给药后对动物的生理观察指标未见影响;对动物的血清生化指标无明显影响;血液学指标检测可见红细胞计数(RBC)、血红蛋白含量(HGB)、红细胞压积(HCT)降低,淋巴细胞百分数(L)增高,网织红细胞计数(Ret)增高等;胸腺和脾脏的脏器重量和脏器系数增大,但各脏器组织结构未见与给药相关的异常改变。反复给药后动物可产生抗体,抗体可中和病毒的活性。结论:大鼠对SeV18 hFGF2/dF肌肉注射给药耐受性良好。     

改良大鼠颈静脉采血法

目的改进大鼠颈静脉采血法,以实现实验大鼠高质量、可循环采血,满足特殊实验要求。方法实验大鼠腹腔麻醉后,采取特殊固定方法,充分暴露颈静脉,使用一次性真空采血管进行采血。结果此法创伤性极低,本实验对60只大鼠进行间隔14 d的采血,一次采血1.8~2.5 m L,成功率在95%以上,对大鼠正常活动无明显影响。真空采血法保障了血清质量,血液检测与分析结果的准确性明显提高。结论在动物福利原则基础上,改进后的大鼠颈静脉采血法具有操作简捷、创伤小、采血量大、血液标本质量高、可循环采血等优点,保障了特殊实验的有效进行。     

心肌梗死大鼠体外培养心肌组织对间充质干细胞迁移的作用

背景:间充质干细胞具有改善心脏功能的潜力,间充质干细胞移植后向心脏归巢的机制尚不完全清楚。目的:观察心肌梗死大鼠体外培养的心肌组织对间充质干细胞迁移的作用及可能机制。方法:建立大鼠心肌梗死模型,培养大鼠心脏组织块,密度梯度离心及贴壁筛选法分离培养骨髓间充质干细胞,24只SD大鼠随机抽签法分为心肌梗死组、假手术组及正常对照组。假手术组只穿线不结扎,正常对照组不做任何处理。荧光染料DAPI标记间充质干细胞,利用transwell模型进行共培养,共培养48h。荧光镜下计数迁移细胞数,CD34/CD44免疫细胞化学染色进行迁移细胞定性,免疫荧光检测迁移细胞CXCR4的表达情况,心脏组织切片行免疫组织化学染色检测基质细胞衍生因子1的表达情况并进行平均吸光度分析。结果与结论:心肌梗死组单位视野迁移细胞数显著高于假手术组(P0.05),正常对照组未见迁移细胞。免疫细胞化学染色显示迁移细胞CD44阳性而CD34阴性,符合间充质干细胞特点,其膜受体CXCR4阳性表达。心肌梗死组及假手术组心脏切片基质细胞衍生因子1阳性表达,正常对照组心肌组织不表达基质细胞衍生因子1。心肌梗死组心脏组织基质细胞衍生因子1平均吸光度显著高于其他组别(P0.05)。提示心肌梗死后大鼠心脏组织能促进间充质干细胞迁移,这一效应的实现可能与基质细胞衍生因子1-CXCR4轴的作用有关。     

观察大鼠胚胎心内膜垫发育时的切面选择

心内膜垫 ( endocardialcushion,EC)是整个胚胎心脏发育过程中的重要部位 ,本文旨在研究不同切面时胎龄( ED) 1 1 d～ 1 9d Wistar大鼠胚胎 EC的形态学表现 ,探讨观察大鼠胚胎心内膜垫发育的最佳切面 ,为进一步开展心脏胚胎发育生物学提供形态学基础及技术支持。材料和方法 :实验动物用清洁级 ED1 1 d～ 1 9d Wistar大鼠胚胎 ,同一胎龄各取 1 5只胎鼠 ,将其经 1 0 %中性福尔马林固定 2 4hr,常规石蜡包埋 (融点 5 3℃ )、切片 (厚度 5 μm)后 HE染色 ,光镜下观察。每一胎龄各取 5只胎鼠分别行冠状切、水平切及矢状切。结果 :ED1 1 d E…     

甲状旁腺切除对大鼠离体心脏缺血性挛缩及再灌注冠脉流量的影响

本实验将大鼠双侧甲状旁腺切除,观察其对大鼠离体心脏缺血性挛缩及再灌注冠脉流量的影响。材料与方法Wistar雄性大鼠60只,体重220～280g,随机分成对照组及实验组,每组30只。对照组行假手术,实验组切除双侧甲状旁腺。常规切片及术后三周时血浆钙明显降低(<1.75mmol/L)作为手术成功的指标。术后三周时将大鼠心脏取出,1分钟内以Langendo-     

大鼠实验中几种常用的采血方法探讨

目的探讨大鼠采血方法，提高大鼠实验的成功率。方法采用SD大鼠300只进行采血。腹主动脉采血法：在腹主动脉分叉处向心端1～3mm处为最佳穿刺点。眼眶后静脉丛的采血法：垂直插入内眦并向眼底方向转动以便切开静脉丛。心脏穿刺采血方法：剑突下，以角度25°～30°斜行向上进针刺人皮下，针尖穿过横膈膜继续斜行刺入2．5～3．0cm。尾尖采血方法：手术剪剪去尾尖5～10mm。颈静脉法：第4肋骨水平方向刺人皮肤，向颈静脉进针，胸部表面成角度30°～40°向颈静脉刺入，刺入的深度约5mm。结果腹主动脉采血法：腹主动脉采血78次，成功73次，成功率93．6％。眼眶后静脉丛的采血法：眼眶后静脉丛的采血法成功率89．9％。心脏穿刺采血法成功率82．4％。尾尖采血法成功率94．4％。颈静脉法成功率80．9％。结论腹主动脉采血法适用于取血量大、不易溶血，不损伤器官，不会出现因操作不当造成的气栓与瘀血，有利于病理组织学检查。几种方法各有优缺点，各有注意事项。应根据自己的实验设计和要求来选择不同的采血方式。     

大鼠颈静脉采血体会

抽取大鼠血液是医学研究中一项最基本最常用的技术,常用的大鼠采血方法包括断头取血、腹主动脉取血、剪尾取血、眼眶后取血、心脏穿刺取血及颈静脉取血等,但是大部分取血方法因有创、采血量不够或不能重复采血而影响实验进程和结     

雌激素及其类似物对去卵巢大鼠脑一氧化氮合酶阳性神经元的影响

雌激素类似物(ZR)具有雌激素样作用,通过建立大鼠卵巢切除术(OVX)模型,初步探讨了雌激素(E)、ZR对脑组织一氧化氮合酶(NOS)表达的影响。实验用雌性Wistar大鼠(3月龄,体重240±10g);行卵巢切除术(OVX);假手术组只暴露卵巢而不切除;大鼠在OVX术后两周开始给药(5周,3d一次;im,200μg/只)。动物分组如下:1假手术组 橄榄油;2OVX 橄榄油;3OVX E;4OVX ZR。给药结束后,心脏灌流固定,取脑冰冻切片40μm,进行NADPN-diaphorase(NADPH-d)反应及nNOS免疫组化,进行图像分析。结果表明:NADPH-d酶组化及nNOS免疫组化标记的神经元在嗅皮…     

Counting cells in cerebrospinal fluid collected directly on membrane filters

Accurate enumeration of cells in the cerebrospinal fluid is not feasible with the current method of counting cells in the Fuchs Rosenthal or Neubauer chamber when the count is near normal since the volume of fluid examined is too small. A sample of adequate volume to permit such accurate assessment may be collected from the lumbar puncture needle into a syringe through a ruled membrane filter in a Swinney cartridge. A study of 291 samples shows that the upper limit of the normal cerebrospinal fluid count is 2000 cells/ml (2/cmm) and not 5000/ml (5/cmm) as is currently accepted.     

Spatial Characteristics of Cisterna Magna in Rats and Novel Technique for Puncture with a Stereotactic Manipulator

The actual size and sites for puncture of the cisterna magna in adult Wistar rats were estimated on acrylic models. We proposed a new method for obtaining cerebrospinal fluid from narcotized rats. This method consists in controlled introduction of a needle into the cisterna magna through the skin and/or dura mater above this structure using a stereotactic micromanipulator. Experiments on rats with focal and global cerebral ischemia showed that this technique ensures single and repeated sampling of at least 50 microl liquor not containing blood admixtures.     

Detection of needle puncture to blood vessel using puncture force measurement

Although blood sampling is frequently performed, a system to take blood samples automatically has not yet been developed. In the paper, as a first step towards automatic blood sampling, an examination of an automatic method for puncturing blood vessels is described. The force waveforms produced by puncturing rabbit ear veins were measured. A characteristic peak, possibly associated with the needle piercing the vessel wall, was observed in each waveform of ten successful cases of 14 trials. An algorithm that allowed the detection of this peak was developed, and parameters of a filter to improve the accuracy of the algorithm were determined. Using this algorithm, automatic needle punctures were performed in a rabbit ear vein and then were simulated using the data derived from manual needle puncture on four other rabbits. The results gave 30 fully successful cases of 33 trials of needle puncture and showed that measurement of the puncture force may be important for automatic needle puncture of blood vessels.     

右肋间入路PTCD穿刺点及肝和肝外胆道定位的解剖学研究

目的利用大体标本进行肝上下界所在高度的测量以及部分肝外胆道的定位,以辅助确定穿刺针在体内的活动范围,为临床上经皮肝穿刺胆道引流(PTCD)穿刺点的选择提供参考。方法以肋间隙和肋骨为参照,测量大体标本右腋中线上肋膈角最低点所在位置;以剑突、脐、耻骨联合所在平面为参照,测量肝上下界所在高度;以右锁骨中线与右肋弓下缘交点为参照,测量胆囊底的实际相对位置偏移;以前正中线为参照,测量Calot三角上的3个点到正中矢状面的距离。结果本研究标本中96.67%的右肋膈角下界高于第11肋;一般肝上界最高点不超过剑突高度以上3.20 cm,不低于耻骨联合高度以上27.93 cm,不低于脐所在高度以上14.26 cm;右腋中线上肝下界到剑突距离/肝下界到耻骨联合距离的比值为0.33~1.35;右锁骨中线上肝下界到剑突距离/肝下界到耻骨联合距离的比值为0.15~0.89;前正中线上肝下界到剑突距离/肝下界到耻骨联合距离的比值为0.05~0.79。结论本研究测得的肝上下界体表定位可作为临床工作中的辅助手段;临床可将右肋间入路PTCD的体表穿刺点设计为右腋中线上第11肋间,避免穿刺针误入胸膜腔。     

Frequency of sternal foramen evaluated by MDCT: a minor variation of great relevance

Due to inadvertent cardiac or great vessel injury, sternal foramina may pose as a great hazard during sternal puncture. They can also be misinterpreted as osteolytic lesions in cross-sectional imaging of the sternum. The distribution of these variations differs between populations, but data from Brazilians are scarcely reported. Therefore, this study aimed to verify the frequency of midline sternal foramen and double-ended xiphoid process, as developmental variations, in order to avoid fatal complications following sternal puncture of sternal acupuncture treatment. A total of 114 chest computed tomograms were evaluated. The frequency of midline sternal foramen in a complication risk bearing feature is of approximately 10.5 %. The double-ended xiphoid process was present in 17.5 %. We conclude that sternal acupuncture should be planned in the region of corpus—previous CT should be done to rule out this variation. Furthermore, we strongly recommend the acupuncture technique which prescribes a safe superficial-oblique approach to the sternum.     

Effect of Zingiber officinale on liver oxidative status and biochemical parameters in rats exposed to paraquat

Oxidative stress caused by reactive oxygen species is one of the major pathogenesis of important diseases of animals and human. Paraquat is widely used as herbicide. The toxicity of paraquat is through induction of oxidative processes in biological systems. Biochemically, this herbicide interferes with intracellular electron transfer system leading to the formation of superoxide anion. Zingiber officinale (ginger) is widely used as a spice and medical treatment for various diseases. The objective of this study was to assess the effect of different levels of ginger extract on antioxidant status and serum metabolites of rats. Sixty male albino Wistar rats were divided in six groups as follows: control (saline); ginger; paraquat; paraquat with 100, 200, and 400 ginger. After 30 days of treatment, the blood was collected by cardiac puncture. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST), reduced glutathione (GSH), and lipid peroxidation were estimated. In paraquat-treated group, the serum levels of ALT, AST, and malondialdehyde (MDA) were markedly raised. Administration of ginger extract with paraquat reduced the serum levels of ALT, AST, and MDA. Hepatic SOD, CAT, GPx, GST, and GSH activities were significantly decreased in paraquat-treated group compared to those of control. However, concurrent administration of paraquat with all concentrations of ginger extract had the opposite effect, where it increased the hepatic SOD, CAT, GPx, GST, and GSH activities near to control. The present study demonstrates that administration of ginger extract to rats modulates the antioxidant enzymes and suggests a possible adaptive mechanism to counteract oxidative stress situation.     

A new method for bone marrow cell harvesting

To minimize contamination of bone marrow cells (BMCs) with T cells from the peripheral blood, a new "perfusion method" for collecting BMCs is proposed using cynomolgus monkeys. Two BM puncture needles are inserted into a long bone such as the humerus, femur, or tibia. One needle is connected to an extension tube and the end of the tube is inserted into a culture flask to collect the BM fluid. The other needle is connected to a syringe containing 30 ml of phosphate-buffered saline. The solution is pushed gently from the syringe into the medullary cavity, and the medium containing the BM fluid is collected into the culture flask. There is significantly less contamination with peripheral blood, determined from the frequencies of CD4(+) and CD8(+) T cells, when using this method (<6%) than when using the conventional method (>20%) consisting of multiple BM aspirations from the iliac crest. Furthermore, the number and progenitor activities of the cells harvested using this "perfusion method" are greater than those harvested using the conventional aspiration method. This perfusion method was carried out 42 times using 15 cynomolgus monkeys, and no complications such as pulmonary infarction or paralysis were observed. These findings suggest that the "perfusion method" is safe and simple and would be of great advantage in obtaining pure BMCs, resulting in a less frequent occurrence of acute graft-versus-host-disease in allogeneic BM transplantation.     

X射线引导建立大鼠椎间盘退变模型

背景:目前国内外对于椎间盘退变的造模方法较多,但存在定位不精准问题.目的:对比X射线引导下与非X射线引导下建立大鼠椎间盘退变模型的效果.方法:取9只成年SD大鼠,按照随机数字表法分3组,每组3只,空白组不进行任何操作;对照组手动定位尾椎Co6/7椎间盘,用注射器针头进行垂直穿刺,并均匀缓慢注入无水乙醇,建立椎间盘退变模...     

Measurement of extravascular lung water in hemodialysis patients using blood ultrasound velocity and optical density dilution

In hemodialysis patients, volume homeostasis is an important clinical problem. The aim is to have patients at an ideal "dry weight" postdialysis, but current methods for accurately measuring dry weight are disappointing. Krivitski et al. (ASAIO J 1998;44:M535-M540) have described a novel technique whereby extravascular lung water (EVLW) may be measured using blood ultrasound velocity and electrical impedance dilution. They have tested this method in animals and achieved agreement between obtained versus gravimetric measurements. Isotonic saline is used as a nondiffusible indicator and hypertonic (5%) saline is used as a diffusible indicator. By injecting these solutions and following their transits through the cardiopulmonary circulation, a theoretic basis for the calculation of EVLW may be derived from the cardiac output, the water transferred to blood, the amount of sodium chloride moved from blood to lung, and the increase in blood osmolality measured at the moment of osmotic equilibrium. We have used this new technique to measure EVLW for the first time in humans in 18 stable hemodialysis patients with no cardiac problems. Measurements were carried out twice in each patient, the first early in dialysis, the second toward the end of a dialysis session where fluid removal took place. The values for EVLW were 260+/-49 ml early in dialysis and 230+/-48 ml late in dialysis. This fall of 30+/-45 ml was statistically significant (p = 0.011). EVLW normalized to body weight was 3.29+/-1.0 ml/kg early and 3.02+/-1.04 ml/kg late in dialysis, a nonsignificant difference (p - 0.073). The normalized EVLW values are almost identical to those obtained in animals (3.1+/-1.4 ml/kg) by Krivitski et al. (see above). We conclude that this new technique can conveniently and noninvasively give an estimate of EVLW in hemodialysis patients. The clinical value of this measurement has now to be determined.     

Redistribution of cardiac output after hypothalamic lesions and hemorrhage

Summary The effects of ablation of the anteroventral portion of the third cerebral ventricle (AV3V) on cardiac output and distribution of regional blood flows were determined in conscious rats using 15 m radiolabelled microspheres before, and 2 min and 15 min after hemorrhage (n=11 for each group). Prior to hemorrhage, cerebral blood flow was significantly greater (216±30 ml/min/100 g), and cerebral vascular resistance was lower (0.60±0.09 mm Hg/ml/min/ 100 g) in rats with AV3V lesions than in controloperated animals (132 ±16 ml/min/100 g; 0.92+0.1 mm Hg/ml/min/100 g, respectively), while mean arterial blood pressure, cardiac output, and regional blood flow to other organs were similar. Less blood was withdrawn from animals with AV3V lesions (4.4 ±0.6 ml) than from control-operated rats (6.0±0.5 ml) to reduce blood pressure to approximately 65 mm Hg. Hemorrhage decreased cerebral vascular resistance in control-operated animals (0.52±0.07 mm Hg/ml/min/100 g), but not in rats with AV3V lesions (0.48±0.1 mm Hg/ml/min/100 g). Cardiac output and regional blood flow to other organs were similar between rats with AV3V lesions and controloperated animals following hemorrhage. These data demonstrate that electrolytic ablation of the AV3V region results in a selective increase in cerebral blood flow and decreased cerebral vascular resistance, but does not alter the reflex changes in regional blood flow evoked by hemorrhage.