骨肉瘤细胞特异性结合短肽的筛选

目的：获得与骨肉瘤细胞株os-732特异结合的短肽，作为骨肉瘤靶向治疗的先导化合物。方法：以骨肉瘤细胞os-732为靶细胞，成骨细胞为吸附细胞对噬菌体12肽库进行差减筛选，用细胞ELISA、免疫组化鉴定阳性噬菌体克隆并测序。结果：经三轮筛选，从随机挑选的20个噬菌体克隆中得到9个特异性与骨肉瘤细胞os-732结合，而不与正常成骨细胞结合的阳性克隆。但其氨基酸序列无同源性。结论：得到多个序列不同的特异性结合骨肉瘤的噬菌体克隆，提示骨肉瘤细胞表面结构复杂，具多个骨肉瘤抗原表位。本实验获得的短肽具有一定的亲合力和肿瘤特异性，为针对不同位点的靶向药物设计提供了实验依据。     

乳腺肿块的超声表现与病理对照研究

目的研究超声对乳腺肿块的鉴别诊断价值及其病理基础。方法选取由超声诊断，并经手术病理证实的74例乳腺肿块，分析其二维超声的形态、边缘轮廓、包膜、边界及内部回声、肿块后方回声及侧方声影等表现特征，结合肿块内部及周边的血流情况，将其分为良恶性，并与病理结果对照。结果二维超声诊断乳腺肿块良恶性的准确率为81．2％，二维超声联合彩色多普勒超声及能量多普勒超声后诊断乳腺肿块良恶性的准确率提高为89．1％。结论二维超声结合彩色多普勒超声及能量多普勒超声检查，可提高乳腺良恶性肿块的诊断正确率。     

儿童缺氧缺血性脑病的影像学和脑电图检查结果的对照及意义

目的：观察影像学和脑电图对儿童缺氧缺血性脑病的应用对照及意义。方法：对65例缺氧缺血性患儿作影像学和脑电图检查。结果：脑电图正常6例（9．23％），轻度异常13例（22．03％），中度异常37例（61．03％），重度异常9例（16．94％），脑电图异常率为90．77％。脑电图异常表现为弥漫性慢波增多和局限性癎性放电。影像学异常44例，异常率为67．69％。结论：影像学和脑电图检查对儿童缺氧缺血性脑病的诊断及治疗有一定的帮助。     

Directed differentiation of Balb/C mouse embryonic stem cells into pancreatic islet-like cell clusters in vitro: observation by atomic force microscope]

OBJECTIVE: To observe the morphological changes of Balb/C mouse embryonic stem cells following directed differentiation into pancreatic islet-like cell clusters (PICC) in vitro using atomic force microscope (AFM). METHODS: Balb/C mouse embryonic stem cells were first cultured into embryonic bodies (EBs) and allowed to differentiate spontaneously for 4 days. The cells were then transferred to gelatin-coated dishes for the EBs to attach and spread on the tissue culture plates, in the course of which a series of cell growth factors such as basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1) and nicotinamide were added into the culture medium at specific time points to induce directed differentiation of the stem cells into PICC. Immunocytochemistry was employed to detect the cells positive for insulin and glucagon, which were observed with AFM. RESULTS: The embryonic stem cells developed into cell clusters of different sizes, in which the cells were tightly arranged. Islet B cells were numerous in the center of clusters and darkly stained, but fewer in the peripherals with lighter stains. Islet A cells expressing glucagon were relatively fewer in the cell clusters, found mainly in the peripherals. Scanning of the insulin-positive clusters by AFM revealed large quantity of tissue fibers resembling nerve fibers that formed a reticular structure in disorderly arrangement. Numerous round granules were observed in the cytoplasm of almost identical sizes ranging from 0.5 to 1.0 mum in diameter. CONCLUSION: The cell clusters obtained by directed differentiation are mature in both morphology and function with also well organized structures.     

Internalization of sst2, sst3, and sst5 receptors: effects of somatostatin agonists and antagonists.

The uptake of radiolabeled somatostatin analogs by tumor cells through receptor-mediated internalization is a critical process for the in vivo targeting of tumoral somatostatin receptors. In the present study, the somatostatin receptor internalization induced by a variety of somatostatin analogs was measured with new immunocytochemical methods that allow characterization of trafficking of the somatostatin receptor subtype 2 (sst2), somatostatin receptor subtype 3 (sst3), and somatostatin receptor subtype 5 (sst5) in vitro at the protein level. METHODS: Human embryonic kidney 293 (HEK293) cells expressing the sst2, sst3, or the sst5 were used in a morphologic immunocytochemical internalization assay using specific sst2, sst3 and sst5 antibodies to qualitatively and quantitatively determine the capability of somatostatin agonists or antagonists to induce somatostatin receptor internalization. In addition, the internalization properties of a selection of these agonists have been compared and quantified in sst2-expressing CHO-K1 cells using an ELISA. RESULTS: Agonists with a high sst2-binding affinity were able to induce sst2 internalization in the HEK293 and CHO-K1 cell lines. New sst2 agonists, such as Y-DOTA-TATE, Y-DOTA-NOC, Lu-DOTA-BOC-ATE (where DOTA is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid; TATE is [Tyr3, Thr8]-octreotide; NOC is [1-NaI3]-octreotide; and BOC-ATE is [BzThi3, Thr8]-octreotide), iodinated sugar-containing octreotide analogs, or BIM-23244 were considerably more potent in internalizing sst2 than was DTPA-octreotide (where DTPA is diethylenetriaminepentaacetic acid). Similarly, compounds with high sst3 affinity such as KE108 were able to induce sst3 internalization. In sst2- or sst3-expressing cell lines, agonist-induced receptor internalization was efficiently abolished by sst2- or sst3-selective antagonists, respectively. Antagonists alone had no effect on sst2 or sst3 internalization. We also showed that somatostatin-28 and somatostatin-14 can induce sst5 internalization. Unexpectedly, however, potent sst5 agonists such as KE108, BIM-23244, and L-817,818 were not able to induce sst5 internalization under the same conditions. CONCLUSION: Using sensitive and reproducible immunocytochemical methods, the ability of various somatostatin analogs to induce sst2, sst3, and sst5 internalization has been qualitatively and quantitatively determined. Whereas all agonists triggered sst2 and sst3 internalization, sst5 internalization was induced by natural somatostatin peptides but not by synthetic high-affinity sst5 agonists. Such assays will be of considerable help for the future characterization of ligands foreseen for nuclear medicine applications.     

约氏疟原虫感染不同小鼠免疫分子的应答差异

目的探讨在约氏疟原虫感染过程中不同宿主的免疫应答差异。方法以约氏疟原虫（致死型）感染DBA／2和BALB／c小鼠，计算红细胞感染率；收集感染前和感染后1、3、6、9、12、15、20d小鼠血清，并无菌取出脾脏，培养脾细胞。应用ELISA试剂盒检测小鼠血清中IFN-γ和IL-12水平，并通过Griess方法检测脾细胞培养上清中NO含量。结果DBA／2小鼠的原虫血症峰值水平明显低于BALB／c小鼠，并于感染后第20d左右自愈；BALB／c小鼠于原虫血症达到峰值水平后全部死亡；DBA／2小鼠的IFN-γ和IL-12水平于感染后1d即出现了有意义的升高并持续到第20d；BALB／c小鼠的IFN-γ和IL-12水平仅干感染后1d出现了有意义的升高；DBA／2小鼠NO的产生于感染后3d出现了有意义的升高，第6d达到峰值，而BALB／c小鼠的NO水平始终未见明显升高。结论DBA／2小鼠通过感染早期Th1细胞免疫应答的有效建立能够抑制原虫血症，IL-12是启动并维持Th1细胞免疫应答的关键性细胞因子。     

脑健冲剂对AD模型大鼠行为学及线粒体结构影响的实验研究

目的：研究脑健冲剂对AD模型大鼠行为学及线粒体超微结构的影响。方法：采用D-半乳糖致衰老与β-淀粉样蛋白注射联合所致的AD大鼠模型，服用脑健冲剂，测试大鼠在迷宫中的学习及记忆能力和海马CAl区线粒体体密度、面数密度等形态学指标。结果：脑健冲剂能有效的改善AD模型大鼠的学习、记忆障碍，并能很好地改善线粒体结构的受损程度。结论：脑健冲剂对AD有较明显的治疗作用，为临床应用脑健冲剂治疗AD提供实验依据。     

微透析技术与经脉脏腑相关的研究思考

微透析是生物化学领域一种崭新的活体局部连续采样分析技术，一经问世便被迅速运用于生命科学的各科领域。本文简要介绍了微透析技术的基本原理及其研究进展，并对其在中医经脉脏腑相关研究领域中的拓展应用提出设想和思考。     

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目的探讨乳管镜对伴乳头溢液乳腺疾病的诊断价值。方法利用纤维乳管镜系统对乳头溢液病人进行检查,观察溢液导管及其分支的管腔和管壁的结构,发现并确定病灶位置,记录病变导管的形态特征,对可疑病例定位切除送病检确诊,对2000～2004年879例病人资料进行回顾性分析。结果发现占位性病变551例,镜下诊断为乳头状瘤398例,乳头状瘤病86例,乳腺导管癌67例。手术392例。经病理证实对乳腺癌诊断的灵敏度934%,特异度985%。结论乳管镜可以明确引起乳头溢液疾病的病因、确定病变部位,解决了乳腺导管内疾病不易诊断的难题,可提高早期乳腺癌的发现率。     

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目的　探讨一种更精确而行之有效的测量甲状腺残留量的方法。方法　自 1996～ 2 0 0 1年对 15例原发性甲状腺功能亢进症病人于术前经CT对甲状腺进行体积测量 ,于双侧甲状腺次全切除术后 ,再用增强CT对上述病人残留甲状腺进行体积测量 ,并与术中测得残留量比较。结果　术中测得残留量 2 8～ 13 0 g ,术后CT测量为 3 4～ 16 6g ,二者之间有一定误差 (P =0 0 6 7) ,产生原因主要为术者对残留甲状腺背面气管旁沟处的不规则腺体测量不准确。体积切除率 83 1%～ 97 9% ,体积残留率 2 1%～ 17 9% ,活体组织相对体积质量 (1 0 5 4 8± 0 0 2 0 5 ) g/cm3 ,离体组织相对体积质量 (1 0 6 4 8± 0 0 188) g/cm3 。结论　CT测量更接近实际残留量 ,可重复性强 ,可比性强。体积残留率比腺体残留重量、手术切除率更具有个性化参考意义。