Objective To observe the changes of basement membrane (BM) during carcinogenic change of the atypical hyperplasia of the mammary. MethodsSP immunohistochemical method and two special stain method (Foot and PAS stain method) and the electron microscope were used to observe the changes of the BM. Results BM already changed in atypical hyperplasia grade Ⅱ, and more significantly changed in atypical hyperplasia grade Ⅲ. BM was thinner or thicker somewhere distinctively under the microscope, and some little deletions were observed under the electron microscope in atypical hyperplasia grade Ⅲ, and there was no BM in breast cancer.Conclusion The change procedure of BM was correlated with the changes of the epithelial, atypical hyperplasia. The changes of BM in the atypical hyperplasia are a part of the carcinogenic procedure from the epithelial atypical hyperplasia of the mammary.
Compare the effect of different chemical methods for preparation of acellular nerve scaffold and to provide an effective nerve scaffold for tissue engineering. Methods Fifteen male SD rats of 2 months old, weighing 200-250 g were selected; the bilateral sciatic nerves were harvested and divided into 3 groups according to preparation methods: group A (normal nerve), group B (Sondell method) and group C (optimal method by Triton X-200, SB-10 and SB-16). The morphology was compared by HE, immunohistochemistry and SEM after dispose; the degrees of decellularization, degrees of demyel ination and integrity of the nerve fiber tube were assessed by scoring system. Results HE staining: In group A, thecross section of nerve was roundness, the cell nuclei was dark blue and the myel in sheath was reticular structure. In group B, the axon and cell nuclei disappeared and the structure of endoneurium was destroyed. In group C, the axon and cell nuclei disappeared and the endoneurium become anomal istic round cavum. The immunohistochemistry staining of Laminin: In group A, the myel in sheath was surrounded by basement membrane with dark blue SC nuclei inside. In group B, the myel in sheath and SC nuclei disappeared and the structure of basement membrane destroyed. In group C, the myel in sheath and SC nuclei disappeared and basement membrane become anomal istic round cavum. The immunohistochemistry staining of S-100: In group A, the myel in sheath and SC were brown. In groups B and C, there were no apparent stained myel in sheath. SEM: In group A, the myel in sheath and axon were clear. In group B, the axon and myel in sheath disappeared and basement membrane became anomal istic. In group C, the basement membrane was more regular than that of group B. The degrees of acellularization and demyel ination of groups B and C were superior to that of group A (P lt; 0.05), and the degrees of demyel ination of group C were superior to that of group B (P lt; 0.05). The integrity of nerve fiber tube of group C was superior to that of group B (P lt; 0.05) and similar to that of group A (P gt; 0.05). The total score was the lowest in group C but the qual ity was the best. Conclusion The effect of decellularization of optimal method was similar to that of traditional Sondell method, but the effect of demyel ination and integrity of nerve fiber tube were better than that of traditional Sondell method. And this acellular nerve can be used as a new kind of nerve scaffold material.
Objective To search the most suitable concentration of calcium in the medium for the basement membrane reconstruction in tissue engineering skin in vitro. Methods Composite chitosan tissue engineering skin was prepared according to previous studies. Four groups were included according to the concentrationof calcium (1.00, 1.45, 1.65 and 1.95 mmol/L respectively). After 7 days and 15 days of culture, the histological manifestation of basement membrane in tissue engineering skin was observed by hematoxylin amp; eosin staining and PAS staining, and collagen Ⅳ of basement membrane was detected immunohistochemicallyatthe dermalepidermal junction. Results This tissue engineering skin shared some histological features of normal skin, including a welldifferentiated stratifiedepidermis and a dense dermis. The epithelium in the group of 1.95 mmol/L calcium differentiated better than those in other groups. PAS staining showing a regularly red dying strap domain at the dermal-epidermal junction. Collagen Ⅳ was positively stained immunohistochemically at the dermalepidermal junction inthe tissue engineering skin. Conclusion The above results suggest that the medium with 1.95 mmol/L calcium should be suitable for the growth of composite chitosan tissue engineering skin and the reconstruction of basement membrane.