异种角膜脱细胞基质和几丁糖载体构建角膜基质层的比较

目的探索异种角膜脱细胞基质和几丁糖为载体构建生物角膜组织的可行性，评价二者的结构、功能和生物相容性。方法l％TritonX-100及冷冻干燥处理获得猪角膜脱细胞基质，网状几丁糖材料制备成似角膜片状形态，将体外培养的兔角膜基质细胞种植于两种载体，体外培养2周，形成细胞载体复合物。对构建的角膜组织进行苏木精-伊红染色、扫描电镜观察；同时将两种生物材料移植到兔角膜基质囊袋内，观察其生物相容性。结果角膜基质细胞在两种载体上皆可黏附生长，并分泌细胞外基质。脱细胞基质载体内细胞形态不规则，位于胶原板层间，形成类似正常组织的角膜基质结构；细胞可在几丁糖网状纤维上黏附并包绕生长。两种载体移植到兔角膜囊袋后，脱细胞基质降解较慢，与宿主角膜组织相融性良好，未发生明显排斥及毒性反应；几丁糖载体降解较迅速，但降解产物诱导较严重的排斥反应。结论异种角膜脱细胞基质保留正常组织的胶原板层结构，并具角膜的韧性和良好的生物相容性，更适于作为体外构建生物角膜的载体，而几丁糖材料则需要在构成结构和性能等方面进一步改进．     

Radial glia development in the mouse olfactory bulb

Radial glia are critical for cell migration and lamination of the cortex. In most developing cortical structures, radial glia, as their name suggests, extend processes from the ventricle to the pia in regular parallel arrangements. However, immunohistochemical labeling from several laboratories suggests that radial glia have a more branched morphology in the olfactory bulb. To investigate the morphology of radial glia in the mouse olfactory bulb we (1) labeled radial glia and olfactory receptor neuron axons at 24-hour intervals by immunohistochemistry; and (2) developed a novel method of generating and applying "nanocrystals" of 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine perchlorate (DiI) to the ventricle surface such that the processes of single olfactory bulb radial glia are labeled in the embryonic olfactory bulb. We examined the structure and interactions of radial glia with ingrowing olfactory receptor neuron (ORN) axons in late embryonic olfactory bulb development. These results showed that olfactory bulb radial glia do not form straight parallel structures as do radial glia in the neocortex but rather have a convoluted trajectory from the ventricle to the bulb surface. Moreover, olfactory bulb radial glia consistently extend tangential branches at the level of the internal plexiform layer. Beginning at embryonic day 17.5, two types of radial glia can be distinguished: type I radial glia have a process that extends from the ventricle into the glomerular layer. These apical processes form highly restricted tufts, or "glial glomeruli" at the same time that ORN axons are forming "axonal glomeruli." In type II radial glia the apical process does not enter the glomerular layer but instead ramifies within the external plexiform layer. The tight spatiotemporal relationship between the glomerulization of radial glia processes and ORN axons during development suggest that radial glia processes could play a role in the formation and/or stabilization of mammalian glomeruli.     

Characterization of an antiserum to synaptic glomeruli from rat cerebellum

Rabbit anti-rat cerebellar synaptic glomeruli antiserum when absorbed with non-neural tissues reacts only with neural tissues when tested by indirect immunofluorescence on tissue sections. Further absorption with forebrain results in an antiserum which detectably reacts only with synaptic glomeruli and soma of Purkinje cells of both rat and mouse. The developmental expression of the synaptic glomeruli antigen(s) parallels the formation of synapses between mossy fibers and granule cells. Immature synaptic contacts do not contain recognizable antigent(s), whereas only at postnatal Day 15 glomeruli become antigen-positive. At this stage antigen in Purkinje cells is no longer carried in their dendrites, but becomes confined to the cell soma. Staggerer mutant mice still express the immature pattern of antigen distribution on postnatal Day 18.     

Induction of proliferation and differentiation of cultured urothelial cells on acellular biomaterials

OBJECTIVE: To determine the optimum conditions for the proliferation of urothelial cells, leading to the confluent coverage of large surfaces of biocompatible membranes, and for their terminal differentiation. MATERIALS AND METHODS: Porcine and human urothelial cells were cultured on different matrices under different growth conditions. Proliferative activity and the viability of cells were evaluated using fluorescent markers for nuclei and cytoplasm. Growth and differentiation were assessed by histological, histochemical and immunohistochemical methods. RESULTS: Under fibroblastic induction and supplementation of 5% fetal calf serum (FCS), urothelial cells showed more proliferation than in other conditions tested. Terminal differentiation of superficial cells was achieved by lowering the concentration of FCS to 1% at the air-liquid interface. CONCLUSIONS: The mitogenic effects of the extracellular matrix content of biological membranes and fibroblastic inductive factors are synergistic with each other, and can compensate for a low FCS concentration and the absence of other additives. Lowering the FCS concentration to 1% inhibits the proliferation of urothelial cells and permits their terminal differentiation.     

In vitro assessment of decellularized porcine dermis as a matrix for urinary tract reconstruction

OBJECTIVES: To assess the potential of Permacol (Tissue Science Laboratories, Swillington, UK), a natural matrix derived from decellularized porcine dermis, as a matrix for urological tissue engineering, and thus to develop an in vitro regimen for assessing the biocompatibility of potential biomaterials before experimentation in animal models. MATERIALS AND METHODS: Urinary tract-derived normal human urothelial (NHU) and smooth muscle (SM) cells were grown in monoculture as autologous cell lines. Permacol was assessed for its ability to support colonization by NHU and SM cells. The failure of the Permacol matrix to be infiltrated by SM cells was further investigated using the highly invasive EJ bladder cancer cell line. RESULTS: NHU cells readily attached and grew as a monolayer on the surface of Permacol. Cells stratified when the culture medium was supplemented with 2 mmol/L calcium. EJ cells initially grew on the surface and subsequently invaded the matrix, while SM cells only colonized the surface of Permacol when cocultured with NHU cells. Cytoxicity, evaluated by contact inhibition and conditioned-medium assays, excluded the presence of soluble toxins in the biomaterial. CONCLUSIONS: We developed a simple, reproducible and rigorous regimen for assessing potential biomaterials in vitro. Applying this system might reduce the use of animals and help to identify causes of potential bio-incompatibility. The inability of SM cells to penetrate the Permacol matrix suggests that required matrix-bound signalling factors are absent, possibly as a result of the procedures used for processing Permacol. Identifying the key regulatory factors that regulate SM cell growth and orchestrate regenerative processes in the urinary tract will be important for developing suitable biomaterials for the bladder.     

Differential activation of glomeruli in the ferret's main olfactory bulb by anal scent gland odours from males and females: an early step in mate identification

Peripheral anosmia was previously found to disrupt sex discrimination and partner preference in male and female ferrets. Here we show directly that volatile anal scent gland odourants from male and female ferrets activated overlapping but distinguishable clusters of glomeruli located in the ventral-caudal portion of the main olfactory bulb (MOB) of breeding ferrets of both sexes. No glomerular activation was seen in the accessory olfactory bulb (AOB). The profile of MOB glomerular activation induced in oestrous females by male anal scents was very similar to that induced by direct contact with a male during mating, and oestrogen treatment failed to alter the profile of glomerular activation induced in ovo-hysterectomized females by male anal scents. In rodents, 'atypical' MOB glomeruli, which have dense acetylcholinesterase (AChE) activity in the neuropil, may be activated by body odours from conspecifics. No such AChE-staining 'atypical' glomeruli were found in the ferret's MOB, suggesting that in this carnivore they do not constitute a subset of MOB glomeruli that respond to body odourants. In ferrets of both sexes, volatile body odourants that are detected by the main as opposed to the vomeronasal-AOB accessory olfactory system may play a critical role in mate identification.     

交联型异体脱细胞真皮基质的研制及动物复合皮移植实验

目的：寻找适合烧伤创面移植的异体脱细胞真皮基质（ａｌｌｏ－ＡＤＭ）。方法：将薄中厚小猪皮采用胰蛋白酸等消化,制成含基底膜的ＡＤＭ,再作组织学等检测和猪复合移植实验。结果：①ａｌｌｏ－ＡＤＭ具有无细胞、无细菌、低毒性等特点。②戊二醛不同交联时间组的ＡＤＭ,与自体薄中厚皮片重叠移植于猪全层皮肤切除的创面上,皮片成活率为１００％,早期观察无排斥反应,外观平整;５ｗｋ时组织学结构完整,炎性反应轻,随交联时     

Bordetella pertussis Clones Identified by Multilocus Variable-Number Tandem-Repeat Analysis

Multilocus variable-number tandem-repeat analysis (MLVA) of 316 Bordetella pertussis isolates collected over 40 years from Australia and 3 other continents identified 66 MLVA types (MTs), including 6 predominant MTs. Typing of genes encoding acellular vaccine antigens showed changes that may be vaccine driven in 2 MTs prevalent in Australia.     

大鼠三叉神经脊束核尾侧亚核Ⅱ层内CB样,PV样阳性结构的分布及？…

本研究用免疫组织化学方法观察了Ｃａｌｂｉｎｄｉｎ Ｄ－２８ｋ（ＣＢ）样和Ｐａｒｖａｌｂｕｍｉｎ（ＰＶ）样胞体、纤维和终末在三叉神经脊束核尾侧亚核（Ｖｃ）Ⅱ层内的公布及它们的突触联系。在光镜下观察到ＣＢ样和ＰＶ样阳性胞体、纤维和终末在ＩＩ层内侧带（ＩＩｉ）最为密集，ＰＶ样阳性神经元的胞体稍大，但数量少于ＣＢ样阳性神经元。在电镜下观察到ＣＢ样或ＰＶ样阳性结构主要形成下列４种突触联系：⑴阳性轴突终末与阳     

Glycogen phosphorylase activity in the olfactory bulb of the young rat

The activity of glycogen phosphorylase, the enzyme that controls glycogen breakdown, was histochemically mapped in the olfactory bulbs of 19-day-old rats. The effect of early odor experience on subsequent olfactory bulb phosphorylase activity was also examined. The highest level of phosphorylase staining in the bulb (and seemingly the highest in the brain) was in the glomerular layer, followed by the external plexiform, internal plexiform, granule cell, and olfactory nerve layers. Virtually no activity was visible in the large output neurons of the bulb, mitral, and tufted cells. Early peppermint odor experience, previously shown to increase metabolic activity in specific glomerular foci as measured by 2-deoxyglucose uptake, had no apparent effect on glomerular-layer phosphorylase activity. In some odor-familiar animals, however, patches of activity were seen in the internal plexiform layer in the area of the bulb where foci of high deoxyglucose uptake are seen in response to peppermint. The patches were directly in line with modified glomerular clusters often seen to underlie foci of enhanced deoxyglucose uptake. The existence of particularly heavy activity in the peripheral third of the glomerular layer, where glycogen-containing modified Schwann cells have been localized, raises the possibility that the glomerular-layer activity is at least partially glial in origin. Finally, because of its rich noradrenaline and serotonin innervation and high density of insulin receptors, the olfactory bulb is proposed as a model system to study the interaction of glycogen/glucose metabolism with neural activity in a relatively well-defined neuronal circuit.