Objective To investigate the method of constructing a tissue engineered epidermis with human epidermal cells and polycarbonate membrane, and to establ ish a tissue engineered epidermis with barrier function which is intended to be the replacing model in vitro of skin irritation test. Methods The tissue engineered epidermis was constructed by using polycarbonate membrane as scaffold and stratified differentiated epidermis derived from human keratinocytes. The tissue engineered epidermis was cultured on an inert polycarbonate filter at the air-liquid interface. After 13 days of culture, the composition and structure of tissue engineered epidermis were observed by HE staining, immunofluorescence staining of keratin 10 (K10) amp; K13, K14, laminin,involucrin, and filaggrin, and transmission electronic microscope. The half maximal inhibitory concentration of a substance (IC50) of SDS was determined in the penetration test of tissue engineered epidermis cultured in the absence (control group) or the presence (experimental group) of l i pid supplement for 18 hours. Results The constructed epidermis was similar to normalepidermis, which was consisted of a proliferating basal layer, differentiated spinous layer, granular layer, and stratum corneum. The IC50 values of tissue engineered epidermis cultured in the control group and experimental group were 0.072% (2.36 mmol/L) and 0.183% (6.00 mmol/L), respectively. Conclusion The tissue engineered epidermis constructed on polycarbonate membrane has normal composition and structure and barrier function corresponding to the normal epidermis.
Objective Using chemically extracted acellular methods to treat extracranial section of the canine whole facial nerve, to evaluated its effects on nerve structure and the removal extent of Schwann cells and myel in. Methods Twenty whole facial nerves were exposed from 10 canines [weighing (18 ± 3) kg]. The extracranial trunk of canine facial nerve and its branches (temporal branch, zygomatic branch, buccal branch, marginal mandibular branch, and cervical branch) were dissected under l ight microscope. Twenty facial nerves were divided into the experimental group (n=12) and control group (n=8) randomly. In experimental group, the nerve was extracted with the 3%TritonX-100 and 4% sodium deoxycholate. In control group, the nerve was not extracted. HE staining and immunofluorescence histological stainings for Hoechst33258, P75, Zero, and Laminin were performed. Results After histological staining, it was found that myel in and Schwann cells were removed from the facial nerve while the basal lamina tube remained intact. The whole canine facial nerves (one nerve trunk and multiple nerve branches) had the similar result. Conclusion The canine whole facial nerve has natural structure (one nerve trunk and multiple nerve branches) by extracted with chemically extracted acellular methods, so it is an available graft for repairing the defect of the whole facial nerve.
ObjectiveTo observe the detection of circulating tumor cells (CTCs) in peripheral blood of patients with gastric cancer, and to investigate the relationship between the CTCs and clinicalpathological features of gastric cancer. MethodsSixty cases of gastric cancer from September 2011 to September 2013 of our hospital were as the research object, and compared with 40 cases of benign gastric disease over the same period. These patients' venous blood were collected, the CTCs counts were determined by using the CellTracks AutoPrep fluorescence scaning system, and the relationship between CTCs and clinicopathological features of gastric cancer was analyzed. ResultsThe detection rate of CTCs in gastric cancer patients was 70.0% (42/60), in control group was 7.5% (3/40). The positive rate of CTCs in peripheral blood of patients with gastric cancer was significantly higher than that of benign gastric disease (P<0.05). The positive rate of CTCs in peripheral blood were no correlated with gender, age, N staging, distant metastasis, tumor size, and vascular invasion (P>0.05), but were correlated with the TNM staging of tumor and degree of differentiation (P<0.05). The cumulative survival rates of 12 months and 18 months after operation in CTCs negtive patients with gastric cancer were higher than that CTCs positive patients (P<0.05). ConclusionsThe detection of CTCs is easy to manage and repeatable. The positive rate of CTCs in gastric patients is higher, which can reflect the progression of tumor and serve as the prognostic index in gastric cance patients.