脑康复冲剂对大鼠软脑膜微循环作用的实验研究

目的：脑康复冲剂临床用于治疗脑血栓血瘀证患者，本实验观察其对在大鼠实验性软脑膜微循环障碍的影响。方法：大鼠预防性给药，用高分子右旋糖酐水溶液静脉注射形成大鼠软脑膜的微循环障碍，观察脑康复冲剂对大鼠微循环的影响。结果：用高分子右旋糖酐造模后，大鼠软脑膜的微循环发生障碍，表现为血液流速减慢、血细胞聚集等，脑康复冲剂组血流速度开始减慢的时间明显比生理盐水对照组延长，且分级数也明显优于生理盐水组，对高分子右旋糖酐造成的血液流态障碍，脑康复组明显好于生理盐水组（P＜0．05），毛细血管网交叉点数开始减少的时间也比生理盐水对照组延长，差异显著（P＜0．01）。结论：脑康复冲剂使大鼠血流速度和血液流态得到改善，具有预防和改善动物脑微循环障碍的作用。     

Kv11.1 channel subunit composition includes MinK and varies developmentally in mouse cardiac muscle

The Kv11.1 (also ERG1) K(+) channel underlies cardiac I(Kr), a current that contributes to repolarization in mammalian heart. In mice, I(Kr) current density decreases with development and studies suggest that changes in the structure and/or properties of the heteromultimeric I(Kr)/Kv11.1 channel are responsible. Here, using immunohistochemistry, we report that total Kv11.1 alpha subunit protein is more abundant in neonatal heart and is distributed throughout both adult and neonatal ventricles with greater abundance in epicardia. Immunoblots reveal that the alpha subunit alternative splice variant, Kv11.1a, is more abundant in adult heart while the Kv11.1b variant is more abundant in neonatal heart. Additionally, MinK channel subunit protein is shown to co-assemble with Kv11.1 protein and is more abundant in neonatal heart. In summary, Kv11.1/I(Kr) channel composition varies developmentally and the higher I(Kr) current density in neonatal heart is likely attributable to higher abundance of Kv11.1/I(Kr) channels, more specifically, the Kv11.1b splice variant.     

脐带血血清中六种微量元素含量的研究

本对100例健康胎儿脐带血血清中六种微量元素（锌，铁，铜，钙，镁，锰）含量进行了测定，结果提示，所测的六种元素含量在男，田性胎儿之间似无明显差异（P＞0．05），与成人比较，胎儿的锌，铁，锰含量高于成人，铜，钙含量低于成人，镁含量与成人相近。     

母亲早产对婴儿的影响及预后

目的 分析研究早产有关的危险因素及早产儿发病及死亡的相关因素。方法 选取我院1995年—2000年12月间早产105例，将早产分为病理性早产(76例)，医源性早产(29例)，与同期足月产对照组110例比较。结果 孕期母亲未作产前检查、胎膜早破、妊高征、胎盘因素、多胎、胎位不正、内外科合并症等同早产有关。早产组产前检查明显低于足月妊娠组(P＜0．05)；早产儿发病率及死亡率明显高于足月儿，早产儿死亡的主要原因是呼吸系统疾患(RDS)、颅内出血、感染。结论 孕期多种因素同早产有关，早产是围产儿发病和死亡的主要原因，提高对早产的认识，倡导围生保健新模式，重视早产的临床相关因素，降低围生儿并发症及死亡率。     

大鼠脊髓损伤后NGF及其受体TrkA在运动神经元及神经胶质细胞表达的变化

为探讨脊髓损伤后运动神经元及神经胶质细胞内神经生长因子(NGF)及其高亲和力受体(TrkA)表达的变化,用改良Allen重击法损伤SCI组动物T12脊髓,按伤后存活时间再将动物分为脊髓损1 d组、2 d组和5 d组。各组动物的脊髓切片经ABC法免疫组织化学染色,用光镜观察TrkA及NGF在脊髓前角运动神经元表达的变化和胶质纤维酸性蛋白(GFAP)及NGF免疫反应阳性胶质细胞的反应性增生程度,并进行图像分析。结果显示:脊髓损伤后前角运动神经元TrkA及NGF的表达随脊髓损伤后动物存活时间的延长逐渐上调;脊髓白质和灰质内尤其是皮质脊髓束内GFAP及NGF阳性胶质细胞明显增生;与此同时,室管膜细胞内亦可见明显的NGF免疫反应产物。上述结果表明,脊髓损伤可刺激脊髓前角运动神经元表达TrkA及NGF,通过自分泌维持受损神经元的存活;损伤部位反应性增生的胶质细胞亦可产生NGF,通过旁分泌作用于脊髓前角运动神经元或皮质脊髓束的轴突末梢,以维持运动神经元的存活及促进皮质脊髓束的再生;适时补充外源性神经营养素或改变损伤局部的微环境将有利于受损脊髓的修复和再生。     

软组织平滑肌肉瘤中p16基因的甲基化检测

目的 探讨软组织平滑肌肉瘤（ＬＭＳ）中p16基因ＩＮＫ４Ａ的甲基化状态及其与p16表达的关系。方法 应用ＭＳＰ法检测３８例软组织平滑肌肉瘤,１０例平滑肌瘤及５例正常平滑肌组织中p16基因ＩＮＫ４Ａ的甲基化状态,用免疫组织化学ＳＰ方法检测p16蛋白表达情况。结果 ３８例ＬＭＳ中９例发生异常甲基化,异常甲基化率为２３.7%（９／３８）。其中,７例p16蛋白表达阴性,２例p16蛋白弱阳性,在p16蛋白表达阴性的ＬＭＳ中,异常甲基化率为５０％（７／１４）。结论 p16基因第一外显子启动子区５‘CpG岛的异常甲基化是导致p16基因失活、蛋白缺如的重要基因外机制,并可能参与肿瘤的发生。     

人TH基因重组腺病毒的构建及生物学活性研究

将人酪氨酸羟化酶cDNA表达盒克隆于质粒型腺病毒载体p△Elsp1A，得到重组质粒pAd－TH。随后用脂质体法将重组质粒pAd－TH和拯救型腺病毒质粒pBHG11一起共转染293细胞，通过体内同源重组生成重组腺病毒AdCMVth，THcDNA重组进入腺病毒E1区并受CMV启动子控制。采用形态学、病毒核酸酶切和PCR／RT－PCR等方法进行鉴定正确。重组腺病毒滴度达到1010pfu／ml。初步结果表明，该重组腺病毒感染MN9D细胞后可使细胞内多巴胺水平增加1倍，显示出明显的TH生物学活性。提示TH重组腺病毒AdCMVth可作为高效的基因转移载体用于帕金森氏病基因治疗。     

听源性惊厥易感性大鼠上丘神经元构筑的研究

用石蜡切片Nissl染色方法，光镜下计数、结合计算机图象分析系统观察、测量惊厥鼠与正常鼠土丘神经元的一些指标．结果显示：（1）在吻例段和中段上丘第Ⅱ层和吻侧段上丘第Ⅲ层的神经元胞体平均直径惊厥鼠显著小于正常鼠，说明惊厥鼠上丘上述部位神经元较小；（2）在吻侧段上丘第Ⅵ层，中段上丘第Ⅰ、Ⅱ层和屋倒段上丘第Ⅴ层，神经元剖面椭圆率惊厥鼠显著地小于正常鼠，说明惊厥鼠土丘上述部位神经元胞体较细长；（3）除第Ⅲ层外，土丘各板层神经元剖面面数密度惊厥鼠都大于正常鼠．本研究结果表明，惊厥鼠的土丘有神经元的形态学变化。这种变化与惊厥鼠惊厥易感性的形成之间是否存在着某种关系，有待深入研究．     

Effects of synergistic reinforcement and absorbable fiber strength on hydroxyapatite bone cement

Approximately a million bone grafts are performed each year in the United States, and this number is expected to increase rapidly as the population ages. Calcium phosphate cement (CPC) can intimately adapt to the bone cavity and harden to form resorbable hydroxyapatite with excellent osteoconductivity and bone-replacement capability. The objective of this study was to develop a strong CPC using synergistic reinforcement via suture fibers and chitosan, and to determine the fiber strength-CPC composite strength relationship. Biopolymer chitosan and cut suture filaments were randomly mixed into CPC. Both suture filaments and composite were immersed in a physiological solution. After 1-day immersion, cement flexural strengths (mean +/- SD; n = 6) were: (2.7 +/- 0.8) MPa for CPC control; (11.2 +/- 1.0) MPa for CPC-chitosan; (17.7 +/- 4.4) MPa for CPC-fiber composite; and (40.5 +/- 5.8) MPa for CPC-chitosan-fiber composite. They are significantly different from each other (Tukey's at 0.95). The strength increase from chitosan and fiber together in CPC was much more than that from either fiber or chitosan alone. The composite strength became (9.8 +/- 0.6) MPa at 35-day immersion and (4.2 +/- 0.7) MPa at 119 days, comparable to reported strengths for sintered porous hydroxyapatite implants and cancellous bone. After suture fiber dissolution, long macropore channels were formed in CPC suitable for cell migration and tissue ingrowth. A semiempirical relationship between suture fiber strength S(F) and composite strength S(C) were obtained: S(C) = 14.1 + 0.047 S(F), with R = 0.92. In summary, this study achieved substantial synergistic effects by combining random suture filaments and chitosan in CPC. This may help extend the use of the moldable, in situ hardening hydroxyapatite to moderate stress-bearing orthopedic applications. The long macropore channels in CPC should be advantageous for cell infiltration and bone ingrowth than conventional random pores and spherical pores.     

Generation of monoclonal antibodies to cryptic collagen sites by using subtractive immunization

The extracellular matrix (ECM) plays a fundamental role in the regulation of normal and pathological processes. The most abundantly expressed component found in the ECM is collagen. Triple helical collagen is known to be highly resistant to proteolytic cleavage except by members of the matrix metalloproteinase (MMP) family of enzymes. To date little is known concerning the biochemical consequences of collagen metabolism on human diseases. This is due in part to the lack of specific reagents that can distinguish between proteolyzed and triple helical forms of collagen. Here we used the technique of Subtractive Immunization (SI) to generate two unique monoclonal antibodies (MAbs HUIV26 and HUI77) that react with denatured and proteolyzed forms of collagen, but show little if any reaction with triple helical collagen. Importantly, HUIV26 and HUI77 react with cryptic sites within the ECM of human melanoma tumors, demonstrating their utility for immunohistochemical analysis in vivo. Thus, the generation of these novel MAbs not only identify specific cryptic epitopes within triple helical collagen, but also provide important new reagents for studying the roles of collagen remodeling in normal as well as pathological processes.