骨组织工程种子细胞的研究进展

随着骨组织工程研究的进展 ,选择什么细胞作为其种子细胞成为近年来研究的热点。目前 ,骨组织工程应用中种子细胞有五种来源 :骨、骨膜、骨髓、骨外组织和早期胚胎。本文介绍了五种来源种子细胞的研究状况 ,并对种子细胞各自存在的问题及应用前景进行了分析。     

Endocrinology and paracrinology: Roles of growth factors during peri-implantation development

Several growth factor ligand and receptor gene products havebeen shown to play roles during preimplantation mammalian development.Genes for insulin-like growth factors (IGFs), transforming growthfactors (TGFs), fibroblast growth factor (FGF), platelet-derivedgrowth factor (PDGF) and receptors for insulin, IGF, PDGF, TGFand epidermal growth factor (EGF) are expressed by early embryosof several species including mouse, rat, cow and sheep. Rolesof growth factors during early development have been demonstratedby addition of purified growth factors to culture medium orby molecular genetic techniques that interfere with gene expression.In this way, it has been shown that successful development ofthe blastocyst is dependent on the action of epidermal growthfactor (EGF) and leukaemia inhibitory factor (LIF). Recent experimentsshow that both LIF and EGF stimulate secretion of urokinase-typeplasminogen activator (uPA) and gelatinase B/ matrix metalloproteinase-9(MMP-9) in day 7 mouse blastocyst outgrowths. At the same time,tissue inhibitors of MMPs (TIMPs) are also expressed by embryonic,decidual and uterine tissues during the implantation process.It appears that LIF may act directly or indirectly, by inducingthe expression of other cytokines, to regulate the temporaland spatial production and activity of proteases and proteaseinhibitors to create a favourable environment for implantation.     

The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. III. Extraction and purification of RNA and application to slot-blot hybridization analysis.

RNA was extracted from tissues processed by a new fixation and paraffin-embedding method (the AMeX method) and examined by Northern blot analysis and slot-blot analysis. The RNA extraction method for AMeX-processed tissue sections after the deparaffinization step was the same as that for fresh materials. The total amount of cellular RNA extracted from AMeX-processed mouse liver tissue was slightly less than that extracted from fresh tissue. In tissues of malignant lymphoma, the total amount of cellular RNA extracted from 25 sections each 20 microns thick was about 1.6-1.8 micrograms/mm2, regardless of the histological subtype and period of storage. The extracted RNA was moderately degraded, and usually could not be used for Northern blot hybridization analysis. The intensity of ethidium bromide staining and the hybridization signals of RNA extracted from AMeX-processed tissues were usually reduced in comparison with RNA from fresh material, but specific signals could be detected by slot-blot hybridization analysis. We have demonstrated previously that the AMeX method preserves high-molecular-weight DNA and various antigens. Since the present study showed that information on mRNA can be obtained from AMeX-processed tissue, the versatility and usefulness of this method were further proven.     

The cellular structure and lipid/protein composition of adipose tissue surrounding chronically stimulated lymph nodes in rats

To test the hypothesis that chronic immune stimulation of a peripheral lymph node induces the formation of additional mature adipocytes in adjacent adipose tissue, one popliteal lymph node of large male rats was stimulated by local injection of 10 microg or 20 microg lipopolysaccharide three times a week for 6 weeks. Adipocyte volumes in sites defined by their anatomical relations to the stimulated and homologous unstimulated popliteal lymph nodes were measured, plus adipocyte complement of the popliteal depot, and the lipid and protein content of adipocytes and adipose stroma. The higher dose of lipopolysaccharide doubled the mass of the locally stimulated lymph node and the surrounding adipose tissue enlarged by the appearance of additional mature adipocytes. Similar but smaller changes were observed in the popliteal adipose depot of the unstimulated leg and in a nodeless depot. The lipid content of the adipocytes decreased and that of the stroma increased dose-dependently in all samples measured but the changes were consistently greater in the depot surrounding the stimulated lymph node. The protein content of both adipocytes and stroma increased in samples surrounding the stimulated node. We conclude that chronic immune stimulation of lymphoid tissues induces the formation of more adipocytes in the adjacent adipose tissue. These findings suggest a mechanism for the selective hypertrophy of lymphoid-containing adipose depots in the HIV-associated adipose redistribution syndrome.     

Directly bone-anchored implants for fixation of aural epistheses

In cases of missing outer ears, generally, epistheses are attached to eye-glasses which gives insecure attachment and a fairly poor cosmetic result. A new method for stable episthesis attachment is being tested in Gothenburg, Sweden. Threaded, cylindrical titanium implants are inserted with a meticulous technique in the temporal bone of the patient. At the next stage, 3–4 months later, the implants are connected to abutments which are allowed to penetrate the skin, To these abutments a silicone episthesis is attached. Presently, 7 patients who had no outer ear because of congenital disorders, hereditary diseases or status post trauma or tumour surgery have been operated and followed for up to $\text{1}\text{2}$ have been no problems reported with the bone anchorage or the skin penetration.     

Cell cycle arrest of human gingival fibroblasts and periodontal ligament cells by Actinobacillus actinomycetemcomitans: involvement of the cytolethal distending toxin

The cytolethal distending toxin (Cdt) is produced by several Gram-negative bacterial species and causes growth arrest and morphological alterations in mammalian cells. Actinobacillus actinomycetemcomitans, which is involved in the pathogenesis of localized aggressive periodontitis, also produces a Cdt that affects periodontal connective tissue cells. The aim of this study was to investigate in which phase of the cell cycle these cells are arrested and enlarged when challenged with A. actinomycetemcomitans, and to evaluate the involvement of its Cdt. Human gingival fibroblasts and periodontal ligament cells were challenged with A. actinomycetemcomitans extract, or with purified Cdt, and cell cycle analysis was performed by propidium iodide staining and flow cytometry. Cells exposed to an A. actinomycetemcomitans wild-type strain, or to purified Cdt, were arrested in both G1 and G2/M phases, and appeared enlarged compared to the corresponding controls. The cellular enlargement occurred in both G1 and G2/M arrested cells. In contrast, cells exposed to an A. actinomycetemcomitans cdt-knockout mutant strain showed cell cycle phase distribution and size similar to the controls. In conclusion, A. actinomycetemcomitans causes a combined G1 and G2/M growth arrest and enlargement in periodontal connective tissue cells, which is attributed to its Cdt.     

抗癌药用植物猫人参(Actinidia macrosperma)离体再生系统的建立

中药猫人参,又名大籽猕猴桃,是著名抗癌药用植物,其野生资源已近枯竭。本文选用猫人参的茎段(至少带一个节)、叶片作为外植体,筛选出了适合猫人参从启动、快繁直至生根等各个阶段的最适培养基,在实验室建立了程序简单、重复性好的再生系统,实现了猫人参组培苗的快速繁殖。以茎段作为外植体效果较好,既容易消毒,同时又能很快诱导腋芽萌发;以叶片作为外植体,能成功诱导出愈伤组织,但此愈伤组织的分化能力极低。以茎段作为外植体获得的无菌培养物在不同发育阶段适宜的培养基组成:外植体启动培养基为MS+BA 5 μmol/L+NAA 1 μmol/L;最适增殖培养基为MS+BA2 μmol/L+GA 1.5 μmol/L+ZT 4μmol/L,35 d繁殖系数为9-10;最适生根培养基为1/2MS+IBA 2μmol/L,生根率达1 00％,培养35 d,平均苗高6.28cm,平均移栽成活率为91％。     

Reproduction and thermoregulation in Peromyscus: Effects of chronic short days

Seven groups of laboratory reared Connecticut P. leucopus were exposed to a 9L:15D photoperiod for either 6, 9, 12, 20, 25, 32, or 36 weeks. An eighth group was maintained on 16L:8D. During the final 6 weeks all mice were cold exposed (13°C). Substantial reproductive regression occurred in females following 6 weeks exposure to 9L:15D relative to 16L:8D animals. In males, a slight decrease in testicular weight occurred following 6 weeks on 9L:15D; this effect was pronounced by 9 weeks, along with a decrease in seminal vesicle weight. Reproductive recrudescence occurred in females by week 32 and in males by week 36. Between 10 and 12 weeks on 9L:15D mice exhibited increases in nesting, incidence of daily torpor, and presence of the winter molt. These increases persisted through 36 weeks on 9L:15D. An increase in the interscapular brown fat pad occurred between 12 and 25 weeks. Reproductive and thermoregulatory characters respond differently to prolonged exposure to a short day photoperiod in this species.     

Immunohistochemical profile of primary sclerosing Iipogranuloma of the scrotum: Report of five cases

Five cases of primary sclerosing scrotal lipogranuloma were examined histologically and immunohistochemically. Every case lacked a history of injection or trauma, and revealed Common histologicat features; a typical granuloma composed of epithelioid cells and multinucleated giant cells, and inflammatory infiltrates of eosinophils, lymphocytes and macrophageimonocytes in the interstitium. lmmunahistochemistry disclosed the epithelioid cells and multinuclaated giant cells of the granuloma to be monocytetr in nature, as bath types of cells were positive for lyso-yme, α-1-antltrypin, α-1-antichymotrypsin, and KP-1. In the interstitium, KP-1 positive monocytes, L-26 positive B lymphocytes, UCHL-1 positive T lymphocytes and 5–100 protein positive Langerhans-like cells were frequently found. 5100 protein positive cells could not be detected in the granuloma. Primary sclerosing lipogranuloma of the scrotum, therefore, is a peculiar inflammation characterized by granulomas consisting of monocytes and marked tissue eosinophilia of unknown etiology.