Unlike hypoxia, hypoglycemia does not preferentially destroy GABAergic neurons in developing rat neocortex explants in culture

We tested whether hypoglycemia, like hypoxia, would preferentially destroy GABAergic nerve cells in the neocortex. To this end, rat neocortex explants dissected from 6-day-old rat pups and cultured up to a developmental stage approximately comparable to that of the newborn human neocortex, were exposed to hypoglycemia for different periods. Quantitative light microscopic and immunocytochemical evaluation of the cultures demonstrated that hypoglycemia does not preferentially destroy GABAergic but rather non-GABAergic neurons, a finding quite opposite to what was found after hypoxia. Recent biochemical data from other laboratories which seem to support this difference in neuronal vulnerability are discussed. It is concluded that perinatal hypoglycemia may not form such a serious threat with respect to the genesis of epilepsy as does hypoxia.     

组织工程血管管状支架的制备与动态培养在血管组织工程中的应用

目的探索体外构建组织工程血管及细胞种植的方法。方法(1)制备管状支架模具,采用模具成形,冷冻干燥等技术,应用胶原和透明质酸复合体外预构管状支架;(2)分动态种植和静态培养两组进行细胞分层种植,每组12个,应用水溶性磺酸四唑(WST-1)测定,扫描电镜观察,5-溴2-脱氧嘧啶核苷(BrdU)细胞标记鉴定比较两种方法的细胞生长情况。结果(1)成功制备了管状支架,在培养液中约3周可保持管腔形态不变形,不塌陷;2.WST-1测定值在培养6、14、20 d分别为:动态组0.842±0.110、1.124±0.102、1.132±0.112,静态组0.386±0.121、0.489±0.245、0.499±0.265,两组比较差异有统计学意义,电镜和BrdU标记表明动态组细胞排列整齐有序。结论应用模具成形,冷冻干燥技术预构管状支架的方法切实可行,动态培养有助于细胞种植,本研究为组织工程血管的构建方法提供了思路。     

阴道粘膜上皮细胞的分离培养和鉴定

目的 初步建立阴道粘膜上皮细胞体外培养方法,为阴道粘膜上皮研究提供实验模型.方法 取雌性新西兰大白兔阴道粘膜组织小块,胶原酶Ⅳ和胰蛋白酶联合消化分离法收集上皮细胞,接种于角朊细胞无血清培养液中静置培养、传代.动态观察细胞生长增殖情况,扫描和透射电镜观察超微结构,流式细胞仪测定细胞增殖周期,并进行免疫组织化学鉴定.结果 体外培养的阴道粘膜上皮细胞为二倍体细胞,增殖状态良好,细胞间可见桥粒连接,免疫组化角蛋白染色阳性.细胞的超微结构和免疫组化染色均具有上皮细胞特征.结论 在本实验条件下,体外培养的阴道粘膜上皮细胞具有较好的增殖能力,可作为阴道粘膜上皮研究的理想实验模型.     

白细胞介素-15对骨髓增生异常综合征骨髓造血干/祖细胞增殖的影响

目的：观察白细胞介素(IL-15)对体外培养的骨髓增生异常综合征(MDS)患者CD34^ 细胞增殖作用。方法：应用单克隆抗体免疫磁珠分离系统提取MDS患者CD34^ 细胞，以加IL-15组为实验组，不加IL-15组为对照组，进行液体和甲基纤维素半固体集落培养，计算培养后细胞数和CFU—E、BFU—E、CFU—GM、CFU—GEMM等集落数，并用MTT比色法检测IL-15对MDS患者CD34^ 细胞增殖的抑制作用，流式细胞术检测上述培养细胞周期的变异情况。结果：11例MDS对象平均CD34^ 细胞比例、回收率、纯度和富集倍数均达要求，MTT比色法检测IL-15对CD34^ 细胞的增殖作用呈最佳浓度效应，最佳浓度为20μg／L，细胞增殖抑制最低峰值时间为8d。用0μg／L IL-15(对照组)和20μg／L IL-15(实验组)作用MDS CD34^ 细胞，计数显示培养细胞最大增殖倍数和集落形成比率实验组均较对照组明显增加，IL-15作用后各细胞周期G1、S、G2期比例有明显改变，与对照组比较，差异有统计学意义。结论：IL-15对MDS CD34^ 细胞有促增殖效应，与其它造血生长因子具有协同作用，对MDS治疗可能有一定的应用前景。     

支架及无支架条件下构建组织工程软骨的研究进展

支架材料的研究是组织工程的研究热点之一,软骨细胞复合培养给组织工程软骨的构建带来希望,单一的材料诸如胶原、透明质酸、壳聚糖、纤维蛋白凝胶等已经证明可以与软骨细胞复合培养,两种或者多种材料复合可以提高材料的性能,更好地用于组织工程软骨的构建,并满足软骨缺损修复的需要;同样,在无支架情况下应用软骨细胞聚集培养、沉淀培养的方法,可以构建组织工程软骨,并给软骨缺损的修复带来新希望,但目前的研究较少。两者是组织工程软骨构建的两个主要方向。     

非功能性胰岛细胞瘤16例临床分析

目的探讨非功能性胰岛细胞瘤的诊断和治疗方法。方法对近16年间收治的非功能性胰岛细胞瘤16例临床资料回顾性分析。其中位于胰头部4例,胰体尾部12例,瘤体长径平均12cm。结果行肿瘤摘除术6例,胰十二指肠切除术3例,行包括肿瘤在内的远端胰腺及脾脏切除3例,保留脾脏的胰体尾切除术3例,因肿瘤广泛侵浸未能切除1例。治愈性切除率为93.75%(15/16),全组无手术死亡。随访资料显示:11例良性肿瘤患者全部存活,已2～16年。5例恶性肿瘤患者中,1例因肿瘤广泛侵犯未能切除者,术后5个月死于多器官衰竭;1例胰体尾切除并左肝切除者存活2年;胰十二肠切除者2例,分别存活1年和6年,胰体尾切除加脾切除1例已存活12年8个月。结论钡餐,B超,CT,MRI,以及ERCP对该病有诊断价值,手术切除是治疗非功能性胰岛细胞瘤的主要方法,效果良好。     

Monoclonal antibody production to human and bovine 2′:3′-cyclic nucleotide 3′-phosphodiesterase (CNPase): high-specificity recognition in whole brain acetone powders and conservation of sequence between CNP1 and CNP2

Monoclonal antibodies against human and bovine 2′:3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) were generated by fusing FOX-NY myeloma cells with spleen cells from RBF/Dn mice previously immunized with the purified brain antigens. The enzyme isolated from bovine brain was quite basic, with an isoelectric point of 9.71 and both the bovine and human enzymes consisted of a closely spaced doublet at approximately 44 and 46 kDa on SDS-PAGE. Six monoclonals were identified as strongly recognizing the enzyme on both ELISA plates and on immunoblots of whole brain protein. Four monoclonals very weakly cross-reacted with guinea pig myelin basic protein. In contrast with two previous reports, some of our monoclonal antibodies did immunostain 2 or 3 protein bands in peripheral nerve, two bands closely corresponding to those immunostained in central nervous system (CNS) myelin, the Wolfgram protein fraction and in acetone powders of whole brain. Each of the 6 monoclonals reacting strongly on immunoblots recognized the enzyme in from 2 to 5 of the species examined (human, bovine, rat, mouse and rabbit). In addition, all 6 monoclonals that immunostained the enzyme in whole brain, myelin and Wolfgram protein immunoblots recognized both CNP1 (44 kDa) and CNP2 (46 kDa). The two closely spaced protein bands observed on SDS-PAGE and previously stained on immunoblots of CNS CNPase using polyvalent rabbit anti-bovine CNPase antisera, and now different monoclonal antibodies, appear to be immunologically related and to contain highly conserved sequences.     

Elemental composition and water content of myelinated axons and glial cells in rat central nervous system

The distribution of elements (e.g. Na, Cl, K) and water in CNS cells is unknown. Therefore, electron probe X-ray microanalysis (EPMA) was used to measure water content and concentrations (mmol/kg dry or wet weight) of Na, Mg, P, S, Cl, K and Ca in morphological compartments of myelinated axons and glial cells from rat optic nerve and cervical spinal cord white matter. Axons in both CNS regions exhibited similar water content ( 90%), and relatively high concentrations (wet and dry weight) of K with low Na and Ca levels. The K content of axons was related to diameter, i.e. small axons in spinal cord and optic nerve had significantly less (25–50%) K than larger diameter axons from the same CNS region. The elemental composition of spinal cord mitochondria was similar to corresponding axoplasm, whereas the water content (75%) of these organelles was substantially lower than that of axoplasm. In glial cell cytoplasm of both CNS areas, P and K (wet and dry weight) were the most abundant elements and water content was approximately 75%. CNS myelin had predominantly high P levels and the lowest water content (33–55%) of any compartment measured. The results of this study demonstrate that each morphological compartment of CNS axons and glia exhibits a characteristic elemental composition and water content which might be related to the structure and function of that neuronal region.     

自体表皮细胞培养与异体真皮组合应用研究

严重烧伤病人皮肤修复中主要未解决的问题是真皮的替代。动物实验结果表明，异体皮移植后５天，用自体培养表皮细胞膜片覆盖真皮床，１４天后复合皮成活率是８４．６％±２．４％。组织学检查证实表皮已形成了复层结构，可见基底层、颗粒层和角质层。临床应用中，异体皮移植后１０天，去除异体表皮覆盖病人的自体培养表皮，３５天后未见排斥征象，异体真皮促进了培养表皮的分层、成熟和完整，组织学检查证实表皮细胞的边缘清楚，已分化形成颗粒层和角质层，真皮多细胞，已血管化，但表皮嵴缺乏。