Chemical traumatization of adult mouse olfactory epithelium in situ stimulates growth and differentiation of olfactory neurons in vitro

This study demonstrates that ZnSO₄ induced chemical trauma results in an in situ regeneration of the olfactory epithelium which, when maintained in vitro, provides an enriched population of olfactory neurons. Therefore, the ability of the olfactory epithelium to respond to chemical trauma with increased mitotic activity can be used to increase growth of neurons in culture. Tissue obtained from normal or vehicle-treated adult mice produced few olfactory neurons, when maintained in culture, compared to cultures established from tissue following an in situ ZnSO₄ trauma. Maximal neuronal yields were obtained in cultures established from tissue that was removed 4–6 days following chemical trauma. The morphological appearance and the presence of cell specific intermediate filament proteins were used to classify the cell types in these olfactory epithelial cultures. Single cells and aggregates of cells which were immunopositive for keratin, but immunonegative for neurofilament protein and GFAP, were identified as epithelioid. Flattened polygonal cells immunopositive for GFAP were identified as glia. A small population of flattened cells was immunonegative for all of the antibodies used in this study. Cells that had processes were immunonegative for GFAP and keratin. Some were immunopositive for 200 kDa and 160 kDa neurofilament proteins but immunonegative for the 68 kDa neurofilament protein. A few of these cells showed positive immunoreactivity with the olfactory marker protein (OMP) antibody and most likely represented the most mature olfactory neurons in the cultures. This trauma-induced culture model using olfactory tissue from adult mice can serve as a source of CNS neurons for comparison with cultured embryonic neurons.     

Development of intestinal epithelial cell lines using a transgenic mouse

The epithelial cells of the colonic mucosa of the animal have proved impossible to culture using standard tissue culture techniques. Immortalization of adult colonic epithelial cells has been unsuccessful due to the lack of DNA synthesis in these cells once they are isolated from the tissue. Recently an unique transgenic mouse bearing a temperature sensitive mutant of the known immortalizing gene, SV40 large T has become available. The advantage of this mouse is that the SV40 large T gene is expressed in every cell. Active immortalizing protein is produced in each cell at the permissive temperature. We have used colonic mucosa from these mice to initiate cultures of epithelial cells from the colon of adult mice. The cells grow readily at the permissive temperature but die within 7 days at the non-permissive temperature. The methods used to develop these cultures are described.     

足组织缺损的显微修复

１９８４年～１９９４年共收治各种足部组织创伤缺损２３６例，均采用显微外科技术进行组织移植修复，其中单纯皮瓣移植修复１８７例，复合皮瓣移植修复４９例。吻合血管游离移植修复９７例，带血管蒂移位修复１３９例。４例前足缺损者用带肩胛骨的复合皮瓣移植修复，６例足跟严重缺损者用带血管的腓骨复合皮瓣移植修复。经１～１０年随访，皮瓣、复合皮瓣、移植或再植的骨骨各全部成活，愈合良好，足功能基本得到恢复，可行走、负重，１８６例恢复了原工作。认为，恢复足部结构完整与重建足跟、足底的感觉是获得良好功能的关键     

Disposition of 14C-dapoxetine in rats: Complementary experiments with whole-body autoradiographic and tissue dissection techniques

Tissue distribution studies, utilizing whole-body autoradiography and organ dissection techniques, were conducted in male Fischer 344 rats following the oral administration of ¹⁴C-dapoxetine HCl, a potent serotonin reuptake inhibitor. The preliminary study using whole-body autoradiography proved invaluable in locating radioactivity in an organ not usually harvested in a tissue distribution study, namely the preputial gland. Selected organs, based on whole-body autoradiography findings, were dissected from rats and analyzed for radiocarbon content by liquid scintillation counting and for parent drug and N-dealkylated metabolites by extraction and HPLC analysis. Highest concentrations of radiocarbon were observed in the organs of absorption and elimination (ileum, cecum, stomach, duodenum, liver, colon, and kidney) but notable quantities were observed in the lung and preputial and Harderian glands. Most tissues had returned to background radioactive levels 72 h after dosing but persistent concentrations of radiocarbon were present in the preputial gland and liver one week after the single dose of ¹⁴C-dapoxetine. Analysis by HPLC demonstrated the presence of parent drug and N-desmethyl metabolite (nor-dapoxetine) in those organs examined; however, the majority of the radioactivity remained unidentified.     

离心力在体外构建组织工程软骨中的作用

目的探讨离心力对软骨细胞功能表达和组织工程软骨结构的影响。方法采用组织化学和免疫组织化学观察组织工程软骨结构以及Ⅱ型胶原表达情况,应用DMMB分光法测定组织工程软骨硫酸化糖胺多糖(GAG)的含量。结果体外培养2、4、8周时,静态培养的组织较离心培养的组织Ⅱ胶原免疫组织化学染色弱;并且其GAG含量低于离心培养组织GAG含量,各组差异均有统计学意义(F分别为12.3、10.2、9.1,P<0.05)。离心培养的组织工程软骨GAG含量于第4周达到高峰,均值为(7.60±0.79)%。结论离心力刺激软骨细胞分泌GAG和Ⅱ型胶原,并且影响组织工程软骨结构的排列。     

一种兔肠源抗菌蛋白分离纯化及其抑菌活性

目的探讨不同摄食控制方法对荷瘤小鼠生存期的影响。方法摄食控制组分别为每日限食2.0 g(DR)、隔1日断食限食4.0 g(IF)、隔2日断食限食6.0 g(SF)和隔1日断食不限食(IF1)4种方法,正常摄食为空白对照组(AL)。观察各组小鼠的体重变化和平均存活天数。结果DR组和IF组平均存活天数分别为19.52±5.24 d和17.96±4.49 d,比对照AL组15.75±1.83 d明显延长(P<0.001和P<0.01)。结论摄食控制恰当,可明显抑制肿瘤细胞的生长、延长荷瘤小鼠的生存期并提高生存质量。     

Capillary hemangioblastoma: histogenesis of stromal cells

Summary The histogenesis of stromal cells in capillary hemangioblastoma has been the subject of debate. The light and electron microscopic studies of hemangioblastomas presented here showed pericytic and leiomyoblastic features in stromal cells. Cells cultured by the monolayer method showed similar features to those of the original tumors. Immunohistochemical studies for glial fibrillary acidic protein and factor VIII/von Willebrand factor indicated that stromal cells were antigenically distinct from astrocytes and endothelial cells. These findings suggest that stromal cells are closely related to pericytes and smooth muscle cells, and support Rhodin's speculation that pericytes serve as a precursor to smooth muscle cells.     

小鼠肾组织体外培养的研究

目的通过观察胚龄12、14、16天胎鼠和生后1天的仔鼠的后肾组织体外培养的存活发育情况,与同期在体比较,建立能模拟体内生存环境稳定的肾组织培养模型。方法采用体外培养倒置显微镜观察,光镜连续切片技术结合体视学定量分析方法,观察、检测培养后肾组织的存活情况以及培养前后肾脏皮质的发育状况。结果对同一胚（日）龄小鼠后肾组织随培养时间逐渐发育,直至成熟;且随胚（日）龄的增加,肾组织块存活比率逐渐下降,肾脏皮质的发育呈下降趋势。结论小鼠后肾组织体外培养胚（日）龄越小其存活发育情况越好,E12天取材培养的肾组织与在体发育一致,利用微孔膜肾组织培养是一种简便有效的神经组织体外培养方法。     

戊二醛交联提高胶原/壳聚糖真皮支架抗细胞收缩能力的研究

目的 应用交联处理的胶原支架,来减缓其降解过多、易收缩变形、机械性能不足等缺点.方法 采用冷冻-冻干法构建胶原/壳聚糖多孔支架,并通过戊二醛交联构建具有良好抗细胞收缩稳定性和细胞相容性的胶原支架.通过测定不同培养时间各类支架的尺寸变化,研究了戊二醛交联对支架抗细胞收缩能力的影响.结果 戊二醛交联后的胶原/壳聚糖支架未见明显收缩,其细胞活性也始终高于干热交联支架.结论 戊二醛能有效地提高胶原基支架的抗细胞收缩能力,并保持其良好的生物相容性.     

脂肪干细胞/Ⅰ型胶原凝胶复合体的构建及其体内外成骨分化研究

目的探讨利用脂肪干细胞与Ⅰ型胶原凝胶复合构建新型骨组织工程复合体的可行性并对其体内外成骨分化情况进行分析。方法取3月龄日本大耳白兔肩胛部皮下脂肪，Ⅰ型胶原酶消化获得细胞，于体外依次进行骨、软骨、脂肪多向诱导分化鉴定；取第3代细胞与Ⅰ型胶原凝胶复合，于成骨诱导条件下体外培养，相差显微镜、扫描电镜观察复合情况并对其碱性磷酸酶、细胞外基质矿化程度进行检测，同时设立单层贴壁培养细胞作为对照（n=4）；两周后移植于裸鼠皮下，12周后处死动物，依次行放射学、组织学检测观察成骨情况（n=3）。结果（1）所获细胞具备骨、软骨、脂肪多向分化潜能，为多能干细胞。（2）形态学观察显示脂肪干细胞呈三维立体生长状态，均匀、高密度悬浮于Ⅰ型胶原凝胶中。在体外成骨诱导条件下，其碱性磷酸酶活性以及外基质矿化程度均显著高于单层贴壁培养细胞（P〈0．01，n=4）。（3）于裸鼠皮下移植12周后，放射学、组织学检测显示脂肪干细胞．Ⅰ型胶原凝胶复合体成骨明显（n=3）。结论（1）IⅠ型胶原凝胶可显著促进脂肪干细胞的成骨分化。（2）作为一种新型的骨组织工程复合物，脂肪干细胞-Ⅰ型胶原凝胶复合体可应用于骨组织工程，特别是腔隙性骨缺损的修复过程。