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。     

Growth fraction in human brain tumors defined by the monoclonal antibody Ki-67

Summary The monoclonal antibody Ki-67, which reacts with cells in the active part of the cell cycle, was used to evaluate immunocytochemically the growth fraction in 22 primary brain neoplasms. The percentage of labelled cells reflected the histological grade of malignancy of each neoplasms. High percentage of Ki-67-positive cells were observed in one choroid plexus carcinoma (60%), one primary melanoma of meninges (40%), three medulloblastomas (40%–50%), one anaplastic astrocytoma and six glioblastomas (10%–40%). One ependymoma had 7% positive cells. Rare positive cells (1%) were present in one pilocytic astrocytoma and one ganglioglioma. Except one negative case, the meningiomas (five cases) had values of positivity ranging from 1% to 6%. Two acoustic schwannomas were negative. These results suggest that immunocytochemical staining with the Ki-67 may be a useful method for measuring the growth fraction in brain neoplasms.Supported in part by Associazione Italiana Ricerca sul Cancro and Ministero Italiano della Pubblica Istruzione     

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.     

种子细胞双相接种技术促进组织工程骨体外成熟的研究

目的 比较观察双相接种法和静置接种法构建组织工程骨对种子细胞增殖、分化和成骨表达的影响。方法 取健康志愿者骨髓，密度梯度离心法分离骨髓间充质干细胞并进行体外扩增、诱导分化，然后应用双相接种法和静置接种法与脱钙骨基质（DBM）复合构建组织工程骨，体外培养3周，用倒置显微镜、扫描电镜观察细胞密度、形态及基质分泌情况，并在不同时相点测定组织块中DNA含量、碱性磷酸酶（ALP）活性及钙含量变化。结果 倒置显微镜下双相接种法构建的组织块可见大量具有一定折光性的梭形细胞存在于胶原基质中。扫描电镜见双相接种法构建的组织块切面较平坦，孔隙较小，细胞包埋在胶原中，而静置接种法构建的组织块可见细胞、细胞外基质较少。在体外培养过程中，双相接种法组织块中细胞的DNA含量、ALP活性均高于静置接种法，钙含量在培养2周后高于静置接种法。结论 双相接种法是一种高效的组织工程骨构建方法，有利于提高接种效率和促进组织工程骨的体外成熟。     

细胞移植的人嗅鞘细胞培养方法的研究

目的探讨适用于细胞移植的人嗅鞘细胞（OECs）分离与培养的方法。方法无菌条件下取自愿捐献孕5个月流产胎儿的完整嗅球进行嗅鞘细胞提取分离、纯化，并对嗅鞘细胞进行免疫组化染色及纯度检测，观察其形态学特点，检测其免疫组化染色的阳性率。结果本方法分离、培养的嗅鞘细胞大多呈三角形、梭形，有些突起较长，生长活跃，增殖迅速，P75、胶质纤维酸性蛋白（GFAP）免疫组化染色呈阳性，嗅鞘细胞阳性率为95．3％。结论该方法简便易行且不影响细胞功能，适用于细胞移植的研究。     

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。     

产科诊断剂量超声波对培养细胞影响的实验研究

采用模拟在人体中使用的实测超声剂量，对体外培养的Ｌ－９２９株细胞进行辐照，通过细胞回复能力试验，观察回复前后的细胞增殖与抑制。对体外培养的人胚肺纤维细胞经１次及５次辐照，观察了ＤＮＡ及细胞核面积的影响。并通过电镜观察了细胞超微结构的变化。上述实验结果，均提示经辐照后细胞有增殖趋向