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Objective To explore the effect of great saphenous vein interruption on distally saphenous neurocutaneous pedicled skin flap. Methods Fifty-two patients with skin and soft tissue lost on ankle received neoplasty using distally crural saphenous neurocutaneous pedicled skin flap. The patients were divided into two groups: the patients in interruption group (25 patients) were treated with great saphenous vein interruption on distally saphenous neurocutaneous pedicled skin flap, the patients in conventional group (27 cases) were treated without saphenous vein interruption. Results Primary healing: 15 patients (55.56%, 15/27) in conventional group, 21 patients (84.00%,21/25) in interruption group. With effusion: 17 patients (62.96%,17/27) in conventional group, 7 patients(28.00%,7/25) in interruption group. With venous crisis: 10 patients (37.04%,10/27) in conventional group, 2 patients (8.00%,2/25) in interruption group. There was statistical significance between two groups on the above 3 indexes (P 0.05). Conclusions Great saphenous vein interruption could relieve swelling, reduce effusion and have higher primary healing rate in neoplasty using distally crural saphenous neurocutaneous pedicled skin flap compared with the conventional method, which greatly reduce the pain and medical expenses of the patients.
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BACKGROUND:Tissue engineering bone application for repairing critical-size segmental defects is stil in research stage. The ideal construction methods have not yet been found. OBJECTIVE:To review the research literatures on tissue engineering bone scaffold material, its shape and effect on the loading of seeding cells, seek appropriate engineered bone scaffolds which are capable of loading a large number of cells effectively and probably, and provide a new way of repairing segmental bone defects. METHODS:The first author performed a data retrieval of PubMed and Wanfang databases from 1994 to 2013, to search the articles addressing the construction method of tissue engineering bone scaffold, and reviewed the literatures systematical y. RESULTS AND CONCLUSION:A total of 379 references were retrieved, including 161 articles in Chinese and 218 articles in English. According to the inclusion and exclusion criteria, 53 articles were final y involved in the analysis. The analysis results indicated that, the needed volume of bone tissue engineering scaffolds for critical-sized section bone defect reconstruction is big, which needs to load a huge number of seed cells. If there is no suitable forms and shapes for celladhesion, the property of so-cal ed engineered bone is similar to pure artificial bone implants. The effective load of seed cells on engineering bone scaffold material and keeping the activity is the first step in clinical practice, as wel as the important guarantee for loading bioactive seed cells. Hence, a more simple and accurate detection method for loading cellquantity is needed. Looking into the retrieved content, effective load cellquantity and its bioactivity are detected by indirect methods, supporting the effectiveness of cellseeding. Some methods can guarantee the cellquantity and seeding pattern, the real load is unknown as wel as the activity. Fabricating engineering bone scaffold into special form and shape are easy to effective seeding, proliferation and maintaining the biomechanical performance, inducing osteogenesis, and final y detecting the load cellquantity and activity on the scaffold through the simple and direct method.
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BACKGROUND:The effects of engineered bone scaffold containing seeding cels with different shapes to repair bone defect are varied, while the loaded cellquantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE:By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cels on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold. METHODS: (1) Experimentin vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with holow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cels (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and colection, cellquantity was counted, absorbance value was finaly detected and cellactivity was proofed by alkaline phosphatase and alizarin red staining. (2) Experimentin vivo: New Zealand rabbits were randomly and equaly divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancelous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilateraly. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1)Experimentin vitro: The drip of cellsuspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cels per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P 0.05). (2) Experimentin vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cellloading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects.
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ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.
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To construct the eukaryotic vector that expresses the fusion protein of Axud1 and influenza virus hemagglutin HA epitope tag, the total RNA was isolated from the peripheral blood lymphocytes, and reverse transcription reaction was used to amplify the full length of human Axud1 cDNA. PCR product of Axud1 was then amplified using specific primers containing HA epitope sequence, and inserted into eukaryotic expression plasmid pcDNA3.1(+)digested with BamH Ⅰand Xba Ⅰ. The recombinant plasmid was identified by polymerase chain reaction, restriction endonuclease mapping and sequencing, and then transfected into human lung adenocarcinoma SPC-A1 cell lines.The fusion HA-Axud1 protein expression in anti-G418 clones was verified by Western blot. This study might be instrumental in further study of the function of Axud1 protein in tumor cells.