In vitro growth of epithelial cells from mucosa derived from different regions of the gastrointestinal tract

Attempts have been made to culture the mucosa from various parts of the gastrointestinal tract. In this study, using an explant culture method, epithelial cells have been successfully cultured from all major regions of the gastrointestinal tract. The success rate, as judged by outgrowth of epithelial cells for at least 4 weeks, varied with the tissue studied with 19/50 colonic biopsies, 5/11 small intestinal biopsies, 9/12 stomach biopsies and 42/47 gallbladder biopsies yielding outgrowth of epithelial cells. Differentiation of the epithelial cells along the mucus cell pathway could be demonstrated on the monolayer cultures using Periodic acid Schiff or Alcian blue staining. Because the cultures were very heterogeneous and many morphological cell types were present in most cultures, differentiation along the other known differentiation pathways of the gastrointestinal mucosa, such as development of absorptive cells and endocrine cells, could not be excluded.
The problem of bacterial contamination, which has hindered previous studies on tissue from these sites, was overcome by decontaminating the biopsy by soaking in dilute sodium hypochlorite (0.04%).     

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.     

The effect of bacterial melanin on electrical activity of neurons of the substantia nigra under conditions of GABA generation

The changes in spike activity of single neurons of the compact part of the substantia nigra, evoked by nucleus caudatus stimulation under conditions of long-term registration of the single and multiple, isolated and combined actions of GABA, GABA-amide, glutamine, and ethanolamine-O-sulfate (EOS) were studied in albino rats. Inhibition of poststimulus activity under GABA action was recorded and the inhibitory effect of GABA-amide was revealed. Primary excitatory and subsequent inhibitory effects of glutamine in combination with EOS were shown. The subsequent administration of bacterial melanin, synthesized by a mutant culture of Bacillus thuringiensis (BT-M) evoked a clear-cut and prolonged excitatory reaction during all the combined actions of GABA, GABA-amide, glutamine, and EOS. Preliminary administration of BT-M abolished the inhibitory poststimulus effects of GABA, GABA-amide, and EOS, as well as glutamine-induced excitation.     

LRIG1下调原代星形细胞瘤细胞增殖的机制

目的观察LRIGl蛋白表达对表皮生长因子（EGF）促肿瘤细胞增殖作用的影响，探讨LRIGl抑制肿瘤细胞增殖的机制。方法原代培养19例星形细胞瘤细胞，用原位杂交检测LRIGl的表达，用噻唑蓝（MTT）法观察EGF对培养细胞的促增殖作用，并分析培养细胞LRIGl表达和EGF干预后的细胞生长率的关系。结果（75．3±11．6）％的培养细胞表达LRIGl蛋白；EGF促进培养细胞增殖（38．0±14．8）％，EGF促细胞增殖的细胞生长率与LRIGl表达程度呈负相关。结论LRIGl蛋白可能通过抑制EGF—EGFR信号抑制肿瘤细胞的增殖。     

骨髓间充质细胞与速固化磷酸钙支架共培养的实验研究

[目的]观察人骨髓间充质干细胞在速固化磷酸钙骨水泥支架中的增殖和生长情况。[方法]将速固化壳聚糖-磷酸钙骨水泥支架材料预先成形。采用全骨髓法分离人骨髓间充质细胞并在体外对细胞进行培养和扩增。72h首次换液,细胞融合80%传代,取第3代细胞用于实验。将细胞与磷酸钙骨水泥支架复合培养,分别利用四甲基偶氮唑蓝MTT法及扫描电镜等检测细胞在材料中的增殖和生长情况。[结果]用酶标仪检测OD值提示共培养后,支架内的细胞数量逐渐增加。扫描电镜结果显示支架材料内部为多孔结构,孔内见有细胞生长,细胞表面可见分泌颗粒。[结论]速固化壳聚糖-磷酸钙骨水泥对人骨髓间充质干细胞有较好的细胞相容性,细胞可在材料内黏附和增殖,是理想的骨组织工程支架材料。     

深低温冷冻肌腱细胞活性的研究

目的研究深低温冷冻方法对肌腱细胞活性的影响,比较程序性降温和普通深低温冷冻法对腱细胞活性的影响.方法纯种SD大鼠24只（出生21 d）,随机分为3组,取双侧跟腱.新鲜肌腱对照组（A）,常规深低温冷冻组（B）,程序性降温深低温冷冻组（C）.采用相同的方法对3组肌腱细胞进行细胞培养.相差显微镜观察原代和传代后细胞的生长,绘制细胞的生长曲线,考察细胞的活性;对细胞进行成纤维细胞染色、胶原染色和对细胞进行形态观察（扫描电镜）;水解法定量分析细胞培养基中羟脯氨酸浓度的变化,检测细胞合成胶原的能力.结果原代细胞培养时A组细胞的生长速度快于B组和C组（P＜0.01）,C组细胞的生长速度快于B组（P＜0.01）,这种生长速度的差异在细胞传代后消失.细胞的形态学和组织学符合成纤维细胞形态.3组细胞培养基中羟脯氨酸浓度变化的差异无统计意义（P＞0.05）.结论经深低温冷冻处理的肌腱中仍存在具有活性的腱细胞,但数量显著少于新鲜肌腱中活细胞的数量.应用计算机控制程序性慢速降温方法处理的肌腱其活细胞的数量有所提高,但仍低于新鲜肌腱中活细胞的数量.     

兔关节软骨细胞的分离、培养和形态学特征

[目的]探讨兔关节软骨细胞的分离、培养方法,观察单层高浓度培养时细胞表型表达情况.[方法]无菌条件下,从 2周龄新西兰白兔的颞颌关节及四肢关节髁突面削取软骨片,采用机械-酶消化法分离软骨细胞,经台盼蓝拒染计数,将细胞按 1× 106个 /孔接种于 6孔培养板,传代培养,描绘生长曲线.利用相差显微镜及透射电镜观察细胞形态.应用甲苯胺蓝及Ⅱ型胶原免疫组化对细胞进行鉴定.[结果]每克软骨能获取 1 5× 106个软骨细胞,活性率为 95%.培养 2～ 3 d,细胞贴壁、变形,呈多角形; 8 d左右,细胞融合成层.透射电镜观察显示细胞核圆形,有丰富的粗面内质网、高尔基体及分泌的基质成分.甲苯胺蓝及Ⅱ型胶原染色阳性.细胞传至 5代后,出现"成纤维细胞样".[结论]本研究建立了简单易行的软骨细胞分离、培养方法;初代、第 2代细胞生长良好,适合于实验研究;软骨细胞 5代培养后,细胞表型发生改变.     

关于我国发行文化彩票的思考

我国的彩票产业如今正处于高速发展的时期，15年来，全国累计发行的福利彩票和体育彩票约为500亿元人民币。笔者认为，发行文化彩票势在必行，不但可以丰富彩票市场的内容，也将给正处于资金短缺状况的公益文化事业募集资金，使彩票发挥更多的社会效益。     

湿润烧伤膏在眼部整容术中的应用

目的：为减轻眼部整容术后反应，减少瘢痕形成。方法：采取手术后手法按压止血，伤口涂美宝湿润烧伤膏，冰袋冷敷，提前拆线的方法。结果：所有475例手术者，均提前两天拆线，创口Ⅰ期愈合，无明显瘢痕。结论：美宝湿润烧伤膏有促进伤口愈合，减轻瘢痕形成的作用。     

冰片开放血-脑脊液屏障对实验性细菌性脑膜炎治疗影响的研究

目的 探讨冰片开放血-脑脊液屏障（BCB）对实验性细菌性脑膜炎治疗的影响。方法 采用日本大耳白兔作为实验动物，随机分为对照组、冰片组和脑膜炎组。经枕大池注入肺炎球菌悬液建立脑膜炎模型，以冰片作为开放BCB的制剂。以连续静脉注射丙戊酸钠后不同时间点的脑脊液中丙戊酸钠浓度变化作为判断BCB通透性的指标；硝酸镧示踪法观察脑组织表现作为BCB通透性的形态指标。给予脑膜炎动物头孢吡肟或冰片＋头孢吡肟治疗，记录动物存亡情况；取脑脊液（CSF）进行细胞计数及生化检查。取脑组织作常规病理染色，以观察冰片开放BCB对实验性细菌性脑膜炎治疗的影响。结果 给药后对照组CSF中丙戊酸钠浓度基本保持稳定，冰片组CSF中丙戊酸钠浓度于给药后0．5h时即高于对照组并逐渐上升，至6h时达高峰，然后开始下降，14h时浓度仍高于对照组。脑组织超微结构显示，对照组无硝酸镧颗粒通过BCB，脑膜炎组硝酸镧颗粒分布于毛细血管基底膜外，冰片组弥漫性分布于神经细胞间隙和轴突周围。给予头孢吡肟治疗后，脑膜炎＋冰片组动物存活率（9／12，75％）明显高于脑膜炎组（6／18，66．7％）（P〈0．05），但两组间CSF细胞数及蛋白、葡萄糖水平差异无统计学意义。近皮层脑组织HE染色显示，脑膜炎组可见脑组织大量炎症细胞渗出和软化坏死灶,血管“套袖”现象明显，而脑膜炎＋冰片组脑组织内炎症细胞渗出较少，无血管“套袖”现象，无明显坏死软化灶。结论 冰片确实能明显增加BCB的通透性并具有可逆性特点，抗生素联合冰片能改善实验性细菌性脑膜炎治疗效果，为细菌性脑膜炎的治疗提供了新的思路。