Influence of mild hypothermia on vascular endothelial growth factor and infarct volume in brain tissues after cerebral ischemia in rats

BACKGROUND: It has been demonstrated that mild hypothermia has obvious protective effect on both whole and local cerebral ischemia. However, the definite mechanism is still unclear for the brain protection of mild hypothermia on cerebral edema, inhibiting inflammatory reaction, stabilizing blood brain barrier, etc. OBJECTIVE: To investigate the effect of mild hypothermia on the expression of vascular endothelial growth factor and the infarct volume after cerebral ischemia in rats, and analyze the brain protective mechanism of mild hypothermia. DESIGN: A randomized grouping and controlled animal trial. SETTING: Department of Neurology, People's Hospital of Yunyang Medical College. MATERIALS: Twenty adult male SD rats of clean degree, weighing (250±30) g, were provided by the animal experimental center, School of Medicine, Wuhan University. The kits for SP immunohistochemistry were purchased from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. METHODS: The experiments were carried out in the laboratory of Department of Neurology, Renmen Hospital of Wuhan University from May to July 2005. ① The 20 rats were divided randomly into normal temperature group (n =10) and mild hypothermia group (n =10). Models of permanent middle cerebral artery occlusion were established with modified nylon suture embolization. The rats were assessed with the Longa standards: 0 point for without nerve dysfunction; 1 for mild neurological deficit (fore claws could no extend completely); 2 for moderate neurological deficit (circling towards the affected side); 3 for severe neurological deficit (tilting towards the affected side); 4 for coma and unconscious; 1-3 points represented that models were successfully established. The rats of the normal temperature group were fed at room temperature, and those in the mild hypothermia group were induced by hypothermia from 2 hours postoperatively, and the rectal temperature was kept at 34-35 ℃ for 72 hours. ② Measurement of infarct volume: All the rats were anesthetized by intraperitoneal injection overdose sodium pentobarbital 7 days postoperatively, and then the heads were cut down to harvest brain. The brain tissues were placed into -20 ℃ refrigerator for 20 minutes, coronal sections of 2 mm were prepared. The infarct sites were not stained, whereas normal brain tissues were stained as red. The infarct volumes were calculated by using MPLAS-500 multimedia color pathological image&&word analytical system. ③ Counting positive cells of vascular endothelial growth factor protein: The brains were harvested by cutting heads, then coronal sections of 2 mm were prepared. Routine dehydration, hyalinization, wax immersion and embedding were performed, then the detected with SP immunohistochemistry, the kits were purchased from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. The cells whose cytoplasm was yellow-brown were positive ones, a single sample as a unit, peri-ischemic site and ischemic core were selected, and the corresponding sites in controlateral hemisphere were taken as controls. Five visual fields were selected from each site to be observed under microscope, the cells were counted, and the average number of positive cells was calculated in each group. The numbers of positive cells were determined with the image analytical apparatus. MAIN OUTCOME MEASURES: Number of the positive cells of vascular endothelial growth factor protein; Infarct volume of rat brain tissue. RESULTS: All the 20 rats were involved in the analysis of results. ① Number of positive cells of vascular endothelial growth factor protein in brain tissue: It was obviously lower in the mild hypothermia group than in the normal temperature group [(24.02±5.05), (36.07±2.69) cells/high power visual field, P < 0.01]. ② Comparison of infarct volume of brain tissue: After MCAO, it was obviously smaller in the mild hypothermia group than in the normal temperature group [(153.25±23.14), (253.45±36.21) mm3, P < 0.01]. CONCLUSION: Mild hypothermia can inhibit the expression of vascular endothelial growth factor and decrease the volume of cerebral infarction. The inhibition of mild hypothermia on the expression of vascular endothelial growth factor may be one of the brain protective mechanisms.     

解释——护理过程中应重视的一个环节

在护理过程中护士和患者进行频繁接触和互动,护士主要应从以下几点做好解释工作,达到消除患者各种顾虑,改善和融洽医患关系,增强与患者的配合能力,使护理工作顺利进行,杜绝护理差错事故的发生,使患者心情愉快,早日康复.①解释环节对护士的素质要求,护士有高尚的职业道德水准,有扎实的基础知识和丰富的临床实践经验.②解释前,对患者的情况做全面了解.③整个护理操作前解释、操作中指导和操作后嘱咐的内容和要求.④鼓励患者及家属积极提出问题,并能全面、科学地做知识解答.笔者旨在通过撰写此文,引起护理同仁对解释环节的重视,不断提高解释水平的护理质量.     

夜间电子生物阻抗测量装置对阴茎勃起功能障碍的检测

目的 :探讨夜间电子生物阻抗测量装置 (NEVA)在阴茎勃起功能障碍的诊断中的应用价值。方法 :对 4 1例主诉阴茎勃起功能障碍者 (障碍组 ) 19例主诉无勃起功能障碍但有射精障碍者 (无障碍组 )进行NEVA检测。结果 :障碍组夜间试验中的夜间勃起的次数、阴茎勃起的最大体积改变、最长持续时间等客观指标均要差于无障碍组 ,差异有统计学意义 (P <0 .0 1)。结论 :NEVA为非侵入性检测、可鉴别心理性阴茎勃起功能障碍和严重的器质性勃起功能障碍 ,且对勃起功能障碍的定性和定量具有一定的客观性。     

神经电生理检测对多灶性运动神经病诊断价值的初步研究

目的 探讨神经电生理检查在多灶性运动神经病(MMN)中的诊断价值。方法对16例MMN患者及16名健康对照进行运动神经传导速度和感觉神经传导速度检查，记录刺激引出的复合肌肉动作电位的波幅、波宽、面积、位相和时限，进行对比分析，判定是否有运动神经传导阻滞(CB)或暂时性离散(TD)，并有选择性地进行常规肌电图检查。结果16例患者均可见有一根以上运动神经或至少一根运动神经的一个以上部位出现CB或CD。其中13例双上肢正中神经，尺神经出现CB，3例以正中神经、尺神经的远端出现CB首发，随病情进展出现下肢腓深神经CB。仅有2例感觉神经传导速度稍有减慢，波幅略有降低。16例患者神经受累区域以下所支配肌肉肌电图检查见有神经源性损害。结论MMN是一种以远端神经受累为主的不对称性周围神经病，神经电生理检查对诊断和鉴别诊断．MMN起重要作用，CB是MMN的主要神经电生理表现。     

金尔伦治疗急性颅脑损伤的剂量效应研究

目的探讨金尔伦(盐酸纳洛酮)在治疗大鼠液压脑损伤后神经功能恢复和病理损害程度的剂量效应.方法将104只SD大鼠随机分为4组,伤后早期分别腹腔注射0.03 mg/Kg(小剂量组)、0.3 mg/Kg(中剂量组)、3 mg/Kg(大剂量组)金尔伦和等量生理盐水(对照组),连续7 d.结果中、大剂量组动物伤后脑神经功能恢复、脑水肿减轻程度及光、电镜检查显著优于对照组及小剂量组.结论伤后早期使用中剂量和大剂量金尔伦(盐酸纳洛酮)对大鼠液压颅脑损伤有明显的治疗效果.     

粗糙脉孢菌漆酶基因的克隆及在毕赤酵母中的初步表达

采用连续延伸PCR方法克隆到粗糙脉孢菌（Neurospora crassa）漆酶基因，并将其克隆到表达载体pPIC9k，重组质粒经线性化、电激转化Pichia pastoris KM71，部分阳性克隆的PCR结果表明：漆酶基因已整合到巴斯德毕赤酵母染色体上，重组菌经甲醇诱导后3～5d产漆酶量最高，为2-3U／mL。     

Mass spectrometric analysis of the structure of gamma II bovine lens crystallin.

The amino acid sequence of bovine gamma II-crystallin has been verified by a combination of electrospray and fast atom bombardment mass spectrometry. The molecular weight of gamma II, isolated by gel filtration and ion exchange chromatography, was determined to be 20,967 +/- 3 by electrospray mass spectrometry. Another aliquot of gamma II was completely digested by trypsin in a medium of 20% CH3CN and 0.1 M Tris, pH 8.2. The tryptic peptides were separated by reversed phase HPLC and identified by their molecular weights, as determined by fast atom bombardment mass spectrometry (FABMS). The identification of each peptide was confirmed by digesting the peptide further to give new peptides whose molecular weights were also determined by FABMS and related to the proposed amino acid sequences. The data from both types of mass spectrometric analyses were consistent with the sequence previously proposed by Hay et al. (J. Biol. Chem. 1987, 146, 332-338), including threonine at position 119. The FAB mass spectrum of one HPLC fraction suggested that disulfide bonding between Cys 18 and Cys 22 was present in at least half the protein preparation. Whether the Cys 18/Cys 22 disulfide bond was present in native gamma II or was produced during isolation or enzymic digestion could not be determined from these studies. Samples that had been stored for several weeks showed that several of the cysteines had become disulfide bonded. These studies illustrate the power of mass spectrometric techniques to accurately confirm the primary structure of proteins and to identify post-translational modifications.     

复式脉冲低能量ESWL治疗肾结石769例报告

目的探讨复式脉冲HB-V型低能量体外冲击波碎石机治疗肾结石的治疗效果.方法采用复式脉冲HB-ESWL-VG型低能量碎石机治疗直径<2.0 cm的各类肾结石769例，治疗工作电压3～9 kV，平均冲击次数2 300次.结果肾盏结石总粉碎率为97.4%，其中上中盏结石复打率为13.1%，术后3个月排净率为89.4%，下盏结石复打率为17.3%，排净率为81.5%；肾盂结石粉碎率为98.3%，复打率为6.1%，术后3个月排净率为93.0%.结论复式脉冲低能量ESWL治疗肾结石具有治疗成功率高、复打率低、无严重并发症、副作用少之优点.     

兔动静脉短路环法诱导组织工程骨支架材料血管生成的初步研究

Objective To compare the effects of 2 vascular carriers, arteriovenous loop and arteri-ovenous bundle, on inducing angiogenesis in coral scaffold of vascularized tissue-engineered bone in animal models.Methods Thirty-six adult male New Zealand rabbits were randomized into 2 even groups.In group A, an arteriovenous loop (AVL) was formed by microsurgical anastomosis at the proximal ends between the femoral poptiteal artery and vein, and placed in the circular side groove of the coral block (6 mm × 8 mm × 10 mm) .In group B, flow-through vessels bundles of both femoral artery and vein were placed in the side grooves of the coral block.All the implants in 2 groups were wrapped by a micro-porous expand-ed-polytetrafluoroethylene (ePTFE) membrane, and fixed subcutaneously by suturing.Evaluation methods included gross morphological observations, histological examinations, India ink perfusion and vascular casting after 2, 4, 6 weeks.The density of blood vessels was analyzed by the statistical software SPSS 10.0.Results All the corals were encased by newly formed fibrovascular tissues in 2 groups.Ink-stained vessels distributed the surfaces and side grooves, and invaded the interspaces of corals.The degree of vascularization increased over the course of experiment.Blood vessel density demonstrated a significant continuous increase between 2 and 6 weeks after implantation in group A.The mean value of blood vessel density in group A (2 weeks 276.60±4.67, 4 weeks 517.20±10.66, 6 weeks 707.00 ±11.87) was significantly higher than in group B (2 weeks 153.60 ±7.16, 4 weeks 269.40±6.80, 6 weeks 279.20±6.53) (P <0.01).Vascular casting showed that in group A, significant blood vessels sprouted from all areas of the loop, espe-cially at the entrance of the arteriovenous pediele where the small tubes were densely interconnected.In group B, however, no blood vessels sprouted from the arteriovenous bundles and only some small vessels grew from the entrance and exit.Conclusions A vascularized coral model can be constructed by inserting an ar-teriovenous loop or an arteriovenous bundle, useful in vascular bone tissue engineering.The former, however, have stronger abilities to induce angiogenesis than the latter.