AIM: To establish the co-culture mode of tissue-engineered tracheal seeding cells, and compare with the routine culture, so as to provide the fundament of tissue-engineered trachea construction by cell compound materials. METHODS: The experiment was completed at the laboratory, Department of Respiratory Diseases, Tangdu Hospital, the Fourth Military Medical University of Chinese PLA from May 2006 to May 2007.①Three healthy New Zealand male rabbits, weighed (250.00?0.75) g, were used in this study.②Co-culture mode: The tracheal epithelial cells and fibroblasts were isolated and then co-cultured for 7-10 days. The two kinds of cells were distinguished according to their different tolerances to trypsin. Subsequently, A hole received 0.5 g/L trypsinization, cells were suspended in DMEM medium containing 0.05 volume fraction of CO2 and then transplanted into B hole (fibroblasts); again, A hole was added with 2.5 g/L trypsin for digestion, afterwards cells were suspended with K-FSM medium and transplanted into C hole (tracheal epithelial cells). Routine culture: after isolation, tracheal epithelial cells were cultured and fibroblasts were purified, respectively.③The co-cultured cells and the traditionally cultured cells were compared via cell growth curve and cell proliferation detected by 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. RESULTS: ①Cell growth: Co-cultured cells grew well, those in A hole increased intercellular apace after trypsinization, while cells in B hole were shaped as fusiform, without the manifestation of typical road stone-like cells, and cells in C hole were apposite, showing lamellar road stone-like morphology.②Cell growth curve: The growth curve of co-cultured cells was identical with that of cells by routine culture method.③Cell proliferation: MTT detection revealed no significant difference in the proliferation of co-cultured cells and cells cultured by traditional method (P

Congenital anorectal malformation(CAM) is one of the most common colorectal diseases in children. Anoplasty has been the only treatment. Because of the fact that anorectal malformation is constantly complicated with congenital defect in the nerve and musculature, postoperative difficulty in defecation is common. Poor long-term quality of life of these affected children is a burden to the children, family, and society. With the advance of modern medicine, emphasizing the functional recovery after anatomical reconstruction of anorectum and conduction of individualized biofeedback training program to improve the defecatory function and long-term quality of life are the new treatment concepts and important progress. Novel methods for the prevention and treatment of CAM are still under exploration.
Anorectal Malformations ; Anus, Imperforate ; surgery ; Child ; China ; Digestive System Surgical Procedures ; Fecal Incontinence ; surgery ; Humans

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Objective To detect characteristics and the pathogenesis of rhodopsin (RHO) gene mutation in an inbreeding family with autosomal recessive retinitis pigmentosa (ARRP). Methods Peripheral venous blood 5-8 ml was abstracted from 8 members in the inbreeding ARRP family and 10 control individuals. DNA gene group was picked. Extron 1-5 of RHO gene was amplified by polymerase chain reaction (PCR),and the mutation of RHO gene was screened by direct DNA sequence measurement. Results The Gln-344-Arg mutation in the RHO gene was detected in 3 patients with ARRP and homozygotes of the mutation in 3 patients were found. Heterozygous of the mutation was detected in the parent of patients and 1 healthy family member. No mutation of RHO gene was found in 2 healthy family members and 10 control individuals. Conclusions The Gln-344-Arg mutation in the RHO gene may be the pathogenic factor of the ARRP family; the frequency of the mutation of RHO gene may increase in the inbreeding ARRP family.

[Objective]To evaluate the surgical techniques,rehabilitation and the results of internal fixation performed for ankle joint fracture.[Method]One hundred and twenty-six patients with ankle joint fractures were treated with open reduction and internal fracture. According to the Danis-Weber classification,there were 12 cases of type A, 86 cases of type B, 28 cases of type C.The follow-up interview period varied from one year to five years,with an average of eighteen months.[Result]The number of patients whose results was better, good, fair and poor was 65, 44, 11 and 6.The total percent-age of good to better clinical results was 86.51%.[Conclusion]Operative treatment and rehabilitation may provide satisfactory fracture reduction and clinical results for ankle joint fracture.The opportunity selection,correct fracture pattern estimation, proper internal fixation and rehabilitation are important to achieve better long term results.

Objective To observe the behavior of Schwann cell migration and wrapping up of fibres during co-culture of Schwann cells with steroframe of PLA and PLGA fibres. Methods Schwann cells were mixed into 30% ECM gel and co-cultured with PLA non-spinning fibre steroframes and seeded on one ends of PLGA fibres which were treated with collagen,polylysine or ECM. The attachment,growth and migration of Schwann cells were observed under phase contrast microscope and laser scan confocus microscope. Results (1)When Schwann cells were co-cultured with PLA steroframes,most of the Schwann cells kept in the fibre holes and attached on PLA fibres to form B?ngner band likd structures. (2)When co-cultured with PLGA fibers,Schwann cells could attach and migrate on PLGA fibres and the number of Schwann cells were obviously increased when PLGA fibres were treated by ECM. Conclusions (1)ECM gel colud promote Schwann cells attachment,growth and migration on PLA steroframes and PLGA fibers. (2)ECM gel is a good integrating substance for constructing tissue engineering bioartificial nerve.

Objective To investigate the effects of adrenomedullin (ADM) on the expression of nuclear transcription factor-?B (NF-?B) in cultured human retinal pigment epithelial (RPE) cells. Methods Cultured human RPE cells were exposed to ADM with the concentration of 10 -7mol/L or lipopolysaccharide (LPS) with the concentration of 10 ?g/ml for 60 minutes. Nuclear translocation of NF-?B was determined by immunofluorescence staining. Western blotting analysis was used to check the expression of NF-?B protein. Results In cultured human RPE cells that were not stimulated, weak NF-?B positive green flurescence was localized in the cytoplasm. After exposed to ADM or LPS for 60 minutes, there was bright NF-?B nuclear staining in cultured human RPE cells. The results of Western blotting analysis demonstrated that in cultured human RPE cells that were not stimulated, a single weak protein strip with the relative molecular weight of 36?103 identified as immunoreative NF-?B could be observed. Both ADM and LPS could markedly increase the expression of NF-?B protein in cytoplasm. Conclusions NF-?B-dependent signal transduction pathway can be activated in cultured human RPE cells by ADM. ADM could induce NF-?B nuclear translocation and increase the expression of the protein in cytoplasm. NF-?B-dependent signal transduction pathway may play an importantrole in the function of ADM.

0 05). The serum levels of both IL-8 and TNF (P