一氧化氮合酶与缺氧复氧所致神经细胞凋亡及银杏叶提取物的保护作用

目的　探讨一氧化氮 (NO)在缺氧复氧诱导神经细胞凋亡中的作用及中药银杏叶提取物的保护机制。方法　实验使用胎龄 16～ 17日Wistar大鼠的大脑皮层神经细胞进行原代分离培养 ;采用Wright Giemsa染色 ,光镜、透射电子显微镜观察 ;原位末端标记法确立缺氧复氧神经细胞凋亡病理模型 ;应用NADPH d组织化学方法检测神经细胞一氧化氮合酶 (NOS)的表达并用计算机图像分析系统进行定量检测。　结果　缺氧复氧可以使大鼠大脑皮层神经细胞发生凋亡 ,随缺氧时间的延长 ,凋亡细胞数渐多 ,至缺氧 8h复氧 18h达高峰 ;在缺氧 2h(H2 R0 组 )和缺氧 8h复氧 18h(R8R1 8)组中神经细胞NOS表达均显著增高 ,与正常对照组比有显著性差异 (P <0 0 1;P <0 0 5 )。EGB能显著抑制此双时相NOS活性的增强 ,并明显降低神经细胞凋亡率。　结论　缺氧复氧损伤可诱导培养的大鼠大脑皮层神经细胞发生凋亡。NOS表达增强从而使NO产生增加可能是缺氧复氧诱导神经细胞凋亡的机制之一。银杏叶提取物 (EGB)经下调NOS表达活性 ,抑制NO的产生保护培养的大鼠大脑皮层神经细胞免于凋亡。     

Formation of 6-nitro-norepinephrine from nitric oxide and norepinephrine in the spinal cord and its role in spinal analgesia

Spinally released norepinephrine is thought to produce analgesia in part by stimulating alpha(2)-adrenergic receptors, which in turn leads to nitric oxide synthesis. Also, nitric oxide is known to react with norepinephrine in vivo in the brain to form 6-nitro-norepinephrine, which inhibits neuronal norepinephrine reuptake. In the present study, we tested the hypothesis that formation of 6-nitro-norepinephrine occurs in the spinal cord and that intrathecal administration of 6-nitro-norepinephrine produces analgesia by stimulating norepinephrine release. 6-Nitro-norepinephrine was present in rat spinal cord tissue and microdialysates of the dorsal horn and intrathecal space. Intrathecal norepinephrine injection increased 6-nitro-norepinephrine. 6-Nitro-norepinephrine also stimulated norepinephrine release in dorsal spinal cord in vitro. Intrathecal injection of 6-nitro-norepinephrine produced antinociception and interacted additively with norepinephrine for antinociception. Spinal noradrenergic nerve destruction increased antinociception from intrathecally injected norepinephrine, but decreased antinociception from 6-nitro-norepinephrine.These results suggest a functional interaction between spinal nitric oxide and norepinephrine in analgesia, mediated in part by formation of 6-nitro-norepinephrine. Stimulation of auto-inhibitory alpha(2)-adrenergic receptors at noradrenergic synapses decreases norepinephrine release. Paradoxically, alpha(2)-adrenergic agonist injection increases and alpha(2)-adrenergic antagonist injection decreases norepinephrine release in the spinal cord. 6-Nitro-norepinephrine may be an important regulator of spinal norepinephrine release and could explain the positive feedback on norepinephrine release after activation of spinal alpha(2)-adrenergic receptors.     

Reduced Left Ventricular Compliance and Mechanical Efficiency after Prolonged Inhibition of NO Synthesis in Conscious Dogs

Acute inhibition of NO synthesis decreases left ventricular (LV) work and external efficiency, but it is unknown whether compensatory mechanisms can limit the alterations in LV mechanoenergetics after prolonged NO deficiency. Eight chronically instrumented male mongrel dogs received 35 mg kg⁻¹ day⁻¹ of N ^ω-nitro-L-arginine methyl ester orally for 10 days to inhibit NO synthesis. At spontaneous beating frequency, heart rate, coronary blood flow, peak LV pressure, end-diastolic LV pressure and the maximum derivative of LV pressure (d P /d t _max) were not significantly different vs. baseline, whereas LV end-diastolic diameter (32.5 ± 1.0 vs. 37.6 ± 1.4 mm) and LV stroke work (515 ± 38 vs. 650 ± 44 mmHg mm), were reduced (all P < 0.05). The slope of the LV end-systolic pressure-diameter relationship was increased at 10 days vs. baseline (13.9 ± 1.0 vs. 9.6 ± 0.9 mmHg mm⁻¹, P < 0.05), while the end-diastolic LV diameter was smaller at matched LV end-diastolic pressures. At fixed heart rate (130 beats min⁻¹), cardiac oxygen consumption was increased (12.2 ± 1.5 vs. 9.9 ± 1.0 ml min⁻¹), and the ratio between stroke work and oxygen consumption was decreased by 33 ±7 % (all P < 0.05) after NO inhibition. We conclude that sustained inhibition of NO synthesis in dogs causes a decrease in LV work despite an increased contractility, which is most probably due to reduced diastolic compliance and a decrease in external efficiency. Thus, prolonged NO deficiency is not compensated for on the level of LV mechanoenergetics in vivo .     

Biochemical characterization of atherosclerotic plaque constituents using FTIR spectroscopy and histology

The behavior of vulnerable atherosclerotic plaques is believed to be closely related to plaque composition. There is a need for an effective in vivo technique for examining plaque constituent properties. In this study, Fourier transform infrared spectroscopy using attenuated total reflectance (FTIR-ATR) was used to assess and analyze the biochemical properties of human atherosclerotic plaques. FTIR spectra clearly revealed prominent spectral features corresponding to plaque constituents of interest: the 2930 cm(-1) and 2850 cm(-1) peaks (indicating the presence of lipids), the 1730 cm(-1) peak (lipid esters), the 1550 cm(-1) and 1650 cm(-1) peaks (fibrous tissues), and the 1100-1000 cm(-1) broad phosphate peak (calcification). Spectral data examined on a qualitative basis correlated well with both gross tissue anatomy and histologic features. Gross spatial mappings of tissue sections of both lipidic and calcified plaques were performed. Spectra from various regions of the plaques demonstrated the evolution of lipid peaks, fibrous tissue peaks, and the phosphate calcification band within the plaques. Histologic analysis corroborated the spectral findings in this study.     

肝尾状叶脉管构筑的应用解剖

应用55例成人肝剥离标本及45例胎儿、新生儿肝管道铸型,研究了肝尾状叶的鞘系及静脉回流。尾状叶有两个恒定的蒂、接受左、右侧鞘系的双重供应,以左侧为主;而尾状突主要由右后叶鞘系分布。15例肝铸型标本的尾状叶左、右侧动脉形成吻合弓。尾状叶动脉供应形式可分为三种,静脉可分为三型并直接汇入下腔静脉。由于血管吻合的存在,在病理状态下,尾状叶也应是沟通门一腔静脉的桥梁。     

Monocyte chemotactic protein-3 (MCP3) interacts with multiple leukocyte receptors: binding and signaling of MCP3 through shared as well as unique receptors on monocytes and neutrophils

The diversity of monocyte chemotactic protein (MCP)3 target cell types, as well as the capacity of MCP3 to desensitize leukocyte responses to other CC chemokines, suggested that MCP3 may interact with multiple CC chemokine receptors. The purpose of this study is to establish how MCP3 binds and activates monocytes and neutrophils. We show that human monocytes exhibit high-affinity binding for ¹²⁵I-MCP3 with an estimated Kd of 1–3 nM and about 10000 binding sites/cell. The binding of ¹²⁵I-MCP3 to monocytes was progressively less well competed by CC chemokines macrophage inflammatory protein (MIP)lα (Kd = 5–10 nM), RANTES (Kd = 5–10 nM), MCP1 (monocyte chemoattractant and activating factor, or MCAF) (Kd = 60 nM) and MIP1β (Kd > 100 nM). On the other hand, unlabeled MCP3 displaced the binding of radiolabeled MIP1α, RANTES, MCP1 and MIP1β as effectively as the isologous CC chemokines. In agreement with the binding data, pretreatment of monocytes with MCP3 completely desensitized the calcium flux in response to MIP1α and RANTES. However, MIP1α and RANTES failed to desensitize the response of monocytes to MCP3. MCP3 and MCP1 partially desensitized each other's effects on monocytes. These binding and cross-desensitization results suggest that MCP3 binds and signals through other binding sites in addition to those shared with MIP1α, RANTES and MCP1. The unidirectional competition for MIP1β binding and signaling by MCP3 suggests the existence of an as-yet unidentified site for MCP3 shared with MIP1β. The existence of another unique binding site(s) for MCP3 was further shown by the failure of any of the other CC chemokines to compete effectively for MCP3 binding on neutrophils. MCP3 in our study was also the only human CC chemokine that consistently chemoattracted neutrophils. These results suggest that MCP3 is a ligand that can bind and activate a broad range of target cells through receptors shared by other CC chemokines as well as its own receptor.     

弓形虫分离株表膜脂肪酸的色质谱分析

本文采用有机质谱法（ＧＣ／ＭＳ）对弓形虫不同分离株，虫体表膜脂肪酸的种类和含量进行观察分析，实验结果表明弓形虫速殖子细胞膜含有６种主要脂肪酸，范围在Ｃ１４－Ｃ１８之间，分别为肉豆寇酸、棕榈油酸、软脂酸、硬脂酸、油酸和亚油酸，而油酸和软脂酸约占总脂肪酸含量的７０％（５２－８４％）。本室自畸胎儿分离获得的ＳＨ多个虫株的脂肪酸含量及组分与国际标准人源ＲＨ株相一致，而ＳＨ４，ＳＨ（１０）与ＰＰ株，ＲＴ株相似，与平行观察的人和哺乳动物细胞膜的脂肪酸在含量与组分上均有明显差异，为鉴定分离的弓形虫株提供了实验依据。     

高中生问题行为的学生-家长-教师三视角研究

目的：从不同视角分析高中生问题行为的特殊性。方法：在开放式问卷调查的基础上，通过项目分析形成高中生行为调查问卷，随机选取高中生、家长和教师进行正式调查。结果：①在对“是否问题行为”的看法上，学生更关注吸毒、自杀、偷窃等行为：家长更关注离家出走、偷窃、恐吓等；教师更关注作弊、学习没有动力和目标、不具备良好的学习习惯等。②三者都认为发生频率最高的问题行为主要是担心成绩、很少做家务、注意力不集中、没有良好学习习惯等。③学生自我报告问题行为的发生频率较家长高。结论：学生、家长、教师对“是否问题行为”的看法存在较大分歧，但对最普遍存在的问题行为的看法较为一致。     

纤维蛋白原与吸附白蛋白、肝素的新型血管支架材料氧化钛的血液相容性

探索纤维蛋白原(Fibrinogen,FIG)与吸附白蛋白、肝素的新型血管支架材料氧化钛(Titanium Oxide,Ti-O)的血液相容性。(1)研制Ti-O,切割成薄膜;(2)Ti-O薄膜涂层白蛋白和肝素;(3)血小板(platelet,PL)吸附试验;(4)酶联免疫试验测FIG吸附量;(5)动物犬股动脉内植入涂层的Ti-O薄膜与对照试片Ti-O和不锈钢(Stainless steel,SS)薄膜。结果发现:Ti-O完全具有固定白蛋白和肝素的结构与性能,比未涂层的Ti-O能更一步减少PL和FIG的吸附,实验动物体内薄膜6个月后取出扫描电镜观察黏附的PL少,形态无改变,血管内无血栓,优于未涂层的Ti-O,更明显优于SS。Ti-O为N型半导体,不易接受FIG的电荷,并且与血细胞有相似的界面张力,决定生物材料Ti-O有较好的血液相容性。Ti-O对白蛋白、肝素有极好的亲和力是因以化学键相结合,在血中进一步减少FIG和PL被涂层的Ti-O吸附。实验证明涂层的Ti-O有持久和稳定的抗凝血性能。     

The effect of a levonorgestrel-releasing intrauterine device on human endometrial oestrogen and progesterone receptors after one year of use

Thirty-four women bearing a levonorgestrel-releasing intrauterine device, 20 micrograms/day (LNG-IUD-20), for 12-15 months were recruited. Endometrial biopsies were collected during the late proliferative phase of the cycle (on cycle days 10-12) before (control) and after the use of the IUD for 12 months, and assayed for oestrogen receptors (ER) and progesterone receptors (PR). An immunohistochemical technique with the peroxidase-antiperoxidase detection system (PAP method) was employed. D75 and JZB39 were the primary antibodies for ER and PR respectively. The immunostaining semiquantitative analysis was performed with a computerized microscope image processor, and expressed as 'grey value'. Both endometrial ER and PR populations were significantly lower after insertion of the IUD (P < 0.01) than in control biopsies. The intensity of nuclear staining and the percentage of positively stained cells for ER and PR in women with LNG-IUD were each about 50% of those in control biopsies. The results suggested that LNG released locally from the IUD has a depressive action on the ER and PR, which may contribute to the contraceptive effectiveness of this type of IUD and also to the possible causes of LNG-IUD-induced irregular bleeding and amenorrhoea.