Gene expression of transforming growth factor beta receptor II in the epidermis of pathological scar / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the gene expression of transforming growth factor beta receptor II (TbetaR II) in pathological scar.</p><p><b>METHODS</b>Twenty samples of pathological scar were collected from 20 burn or trauma patients hospitalized in the General Hospital of Ji'nan Military Command from 2007 to 2009. Twenty specimens of epidermal layer were obtained from the middle portion and the edge of pathological scars. Twenty normal skin specimens which were located more than 10 cm away from the lesion sites of 20 patients were collected as self-controls. Serum from 1-2 mL whole blood were obtained from each of the 20 patients for second self-control. Eight normal skin specimens from 8 patients without pathological scar, discarded from un-related operations, were also collected as negative-control. Positive expressions of TbetaR II in three different skin specimens were determined with biotin-streptavidin-peroxidase staining. Gene expressions of TbetaR II in all specimens were compared with PCR-single strand conformation polymorphism analysis and gene sequencing. Data were processed with Fisher's exact test.</p><p><b>RESULTS</b>Positive expression of TbetaR II in pathological scar epidermis was lower than that in normal skin specimen of patients with pathological scar or normal skin specimen of patients without pathological scar, and TbetaR II was mainly located in the basal layer of epidermis. Positive expressions of TbetaR II were seldom found in acanthocytes, granular cells, and cuticle or even non-existing. No abnormality of TbetaR II was found in normal skin epidermis or serum samples of pathological scar patients or normal skin epidermis of patients without pathological scar. TbetaR II expressing in 8 specimens of epidermis of pathological scar showed abnormal electrophoresis pattern at poly A fragments hand and loss of one A base in DNA fragment (P = 0.044).</p><p><b>CONCLUSIONS</b>There may he abnormal gene expression of TbetaR II in pathological scar epidermis. Replantation of epidermis of scar may increase the risk of scar recurrence, while replantation of normal skin of patients with scar on wound may not increase the risk of scar recurrence.</p>

Transplantation of hypoxia preconditioned bone marrow mesenchymal stem cells improves survival of ultra-long random skin flap / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Random flap is one kind of the most widely used skin flaps in reconstructive surgery; however, partial necrosis of its distal end remains a significant problem now. The aim of this study was to evaluate the effect of hypoxia preconditioned bone marrow mesenchymal stem cells (HpBMSCs) transplantation on ultra-long random skin flap survival in rats.</p><p><b>METHODS</b>Normoxic bone marrow mesenchymal stem cells (nBMSCs) were cultured under normoxia (20% O2) and HpBMSCs under hypoxia (1% O2) for 48 hours before transplantation. Thirty Sprague-Dawley rats were randomly divided into control group, nBMSCs group and HpBMSCs group with each consisting of 10 rats. Survival area of ultra-long random skin flap on the dorsal of rats was measured seven days after flap surgery and cell transplantation. Cell survival in vivo, microvessel density and vascular endothelial growth factor (VEGF) were evaluated by histological examination and enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Compared with other two groups, flap survival area in HpBMSCs group was significantly larger (P < 0.05). Microvessel density in HpBMSCs group (36.20 ± 8.19) was higher than that in nBMSCs group (30.01 ± 5.68) and control group (17.60 ± 4.19) (P < 0.05). VEGF in HpBMSCs group ((300.05 ± 50.41) pg/g) was higher than those in nBMSCs group ((240.55 ± 33.64) pg/g) and control group ((191.65 ± 32.58) pg/g) (P < 0.05).</p><p><b>CONCLUSION</b>HpBMSCs transplantation improves ultra-long random skin flap survival via promoting angiogenesis of more survival cells.</p>

Effects of silencing inhibitor of DNA binding-1 gene on the growth and invasiveness of adenoid cystic carcinoma cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of inhibitor of DNA binding-1 (Id-1) gene in adenoid cystic carcinoma cell growth and invasion behavior.</p><p><b>METHODS</b>With salivary adenoid cystic carcinoma cell lines ACC-M and ACC-2, dedected Id-1 gene expression was screened with immunofluorescence assay. After Id-1 mRNA knocking-down using small interfering RNA, RT-PCR and Western blot were used to detect the different expressions before and after interference, and the growth of cells before and after interference was deceted using the MTT assay, and the cell invasion ability was checked with the use of Transwell chamber assay.</p><p><b>RESULTS</b>Id-1 were both expressed in the ACC-M and ACC-2, and the expression in ACC-M was higher than that in ACC-2. After Id-1 RNA interference, the growth and invasiveness of ACC-M and ACC-2 were inhibited with the restrained degree in ACC-M much stronger than that in the ACC-2.</p><p><b>CONCLUSION</b>In view of the important role of Id-1 in the behavior of growth and invasion in ACC cell, interfering the expression of Id-1 gene is expected to be a novel and effective means for the treatment of adenoid cystic carcinoma.</p>

Effect of bone morphogenetic proteins-2 on mandibular distraction osteogenesis in rabbits / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the Effect of recombinant adenovirus vectors containing human Bone morphogenetic proteins-2 (Ad-hBMP-2) on the for mandibular distraction osteogenesis (DO) in rabbits.</p><p><b>METHODS</b>Twenty-four New Zealand white rabbits were randomly divided into experimental group, and control group and underwent mandibular distraction osteogenesis. After 5 days latency, the distracters were activated at a speed of 0.5 mm every 12 hours for 7 days, then on the first day in the consolidation period, the distraction gaps of experimental group were injected with 0.2 ml Ad-hBMP2 10(12) pfu/L, while the animals of control group were injected with 0.2 ml Ad-EGFP 10(12) pfu/L. At the 7 th and 28 th day of consolidation period, specimens were obtained, X-ray and histomorphology were performed. The bone density and the quantity of new bone formation in the distraction gaps were observed and compared between the two groups at different consolidation period.</p><p><b>RESULTS</b>Ad-hBMP-2 treated specimens demonstrated an increased amount of new bone formation</p><p><b>CONCLUSIONS</b>Adenovirally-mediated delivery of BMP-2 can locally increase bone deposition during DO, which may potentially shorten the consolidation period.</p>

Ectopic bone induction in vivo after transplantation of skeletal satellite cells from green fluorescence protein transgenic mouse transfected by adenoviral vectors encoding bone morphogenetic protein-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the ability of induced ectopic bone using skeletal muscles satellite cells (SMSCs) from newborn green fluorescence protein (GFP) transgenic mice mediated by Ad-BMP2.</p><p><b>METHODS</b>Transplantation of SMSCs transduced with Ad-BMP2 into back lamb muscles of subfascia in wildtype 129sv mice with a complex of collagen scaffords, then the tissue histologic examination, X ray plain film, fluorescence microscopy were used.</p><p><b>RESULTS</b>Transplantation of SMSCs transfected with Ad-BMP2 into back lamb muscles of subfascia generated ectopic bone formation involving GFP-positive osteoblasts and osteocytes 2 weeks and mature bone formation 4 weeks after transplantation. SMSCs non-transfected with Ad-BMP2 failed to induce ectopic bone formation.</p><p><b>CONCLUSION</b>SMSCs retain differentiation potentitality into osteoblasts in response to Ad-BMP2. They are useful tools for analyzing the process of osteoblast differentiation in vivo after transplantation.</p>

Isolation, identification, culture and bionomics of skeletal muscles satellite cells from green fluorescent protein transgenic mouse in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the green fluorescent protein (GFP) expression and the bionomics of skeletal muscles satellite cells (SMSCs) in vitro in GFP transgenic mouse.</p><p><b>METHODS</b>The newborn transgenic mice were acquired to separate skeletal muscles satellite cells with enzyme digestion method. Cells were cultured and subcultured in vitro. Morphological observation, growth curve were investigated to evaluate the proliferation and differentiation characteristics of skeletal muscles satellite cells, fluorescence microscope was used to observe the GFP expression. The cells were identified by immunocytochemical stain. In the basis of identification of anti-sarcometric actin anti-body, the combination of anti-desmin antibody and DAPI (4, 6-diamidino-2-phenylindole) were used to detect the purification of skeletal muscles satellite cells.</p><p><b>RESULTS</b>Immunocytofluorescence suggested the good retain of GFP fluorescence in skeletal muscles satellite cells. The cells showed strong proliferative ability and they were positive with immunocytochemical stain of anti-sarcometric actin antibody and anti-desmin antibody. The combination of anti-desmin and DAPI stain can be used to determine the purification of SMSCs.</p><p><b>CONCLUSION</b>Skeletal muscles satellite cells cultured in vitro showed strong proliferation and differentiation ability. They are fit to construct the cell bank of tissure engineering and to be a useful tool to explore cells fate after transplantation since these cells retain the expression of GFP.</p>

Preliminary study on transformation of the biological function of the cryopreserved osteoblasts cultured in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To culture and amplify the young rabbit's bone marrow stromal cells (BMSCs) in vitro, and to observe the effect of hypothermia on the cells' growing behavior and biological function.</p><p><b>METHODS</b>BMSCs were acquired from the rabbit' tibia bone marrow and induced to mature osteoblasts in vitro. The cultured cells growing well in vitro were preserved in liquid nitrogen. The anabiotic cells having cryopreserved for 1 week were chosen as the experimental group, and the routine 7th generation as the control group. Their biological function in comparion by the examination of morphological changes, cells' proliferation ability, colone forming ratio, synthesis ability of ALP and protein, mineralized nodes forming ability were observed.</p><p><b>RESULTS</b>As contrast to the control groups, the anabiotic cells also grew and proliferated well in vitro except a little more slowly than before. They had the similar general shape in all the time segments, but a little differences in cells' ultrastructure. The experimental groups also had the typical characters of mature osteoblasts, and high abilities of the synthesis of ALP and proteins. The statistic data showed that these two groups had no significant difference (P > 0.05).</p><p><b>CONCLUSION</b>The cryopreserved osteoblasts had the same biological functions and the similar growing behaviors as before. These results suggest that it is practical to use the cryopreserved osteoblasts for further study on bone tissue engineering.</p>

Experimental study of mandibular periosteal distraction in rabbits / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate a novel technique for new bone formation--periosteal distraction osteogenesis.</p><p><b>METHODS</b>A custom made periosteal distraction device was fixed to bilateral surface of the mandible in three rabbits. Periosteal distraction was performed on the left side of the mandible, the right side of the mandible served as the control. The animals were sacrificed at the end of distraction process. All the specimens were X-rayed and histologically examinated.</p><p><b>RESULTS</b>All three animals survived with no obvious complications. Both in mass specimens and X-rays, there showed new bone formation on the distracted side of the mandible. In histological examinations, there was osteoblast-like cell infiltration and bone tissue formation in the distracted area.</p><p><b>CONCLUSION</b>Periosteal distraction osteogenesis can provide a novel technique for the repair of bone defects.</p>

Experimental study on the phenotype of human embryonic lung fibroblasts transfected with Ad-BMP-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects on the phenotype, especially the mineralization ability of human embryonic lung fibroblasts(HELFs) by transfection with adenovirus vector ecoding human bone morphogenetic protein-2 gene (Ad-BMP-2).</p><p><b>METHODS</b>The HELFs were primarily cultured, then transfected with Ad-BMP-2. The morphologic characteristics of the cells were observed. The cell proliferation, alkaline phosphatase activities, BMP-2 protein expression, and the mineralization ability were detected with the methods of MTT, ALP staining, Western blot, and alizarin red S staining, respectively.</p><p><b>RESULTS</b>After transfection, the shape of HELFs changed from silm spindle to multifigure, the cells became bigger than before. The colonies changed from unilaminar into multilaminar. The proliferation of HELFs was severely inhibited after transfection. An obvious BMP-2 lane was shown in Western blotting. Most cells presented positive in ALP staining, and large number of nacarat mineralized nodes were observed after alizarin red S staining.</p><p><b>CONCLUSION</b>HELFs were capable of transforming into osteoblast-like phenotype, and were endowed with the ability of mineralization while being transfected with Ad-BMP-2.</p>

Experimental studies of human adipose tissue-derived stromal cells transfected with ad-hBMP-2 gene / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the possibility of adipose-derived stromal cells (ADSCs) transfeced by adenovirus containing human bone morphogenetic protein-2 (Ad-hBMP-2) gene and their osteogenic potential.</p><p><b>METHODS</b>ADSCs were obtained from inguinal fat tissue of 4 weeks old SD rats. After exposure to adenovirus containing green fluorescent protein(Ad-GFP), fluorescent microscope was used to observe gene transfection effect once 12 hours. After transfected with Ad-hBMP-2, cytochemistry, immmucytochemistry and Western blot were used to examine the expression of alkaline phosphatase (ALP), osteocalcin (OC) and hBMP-2.</p><p><b>RESULTS</b>After exposed to Ad-GFP 12 hours, 52% ADSCs were observed being transfected and 48 hours later reached 95%. The double number time belonged after transfecting with Ad-hBMP-2, and cytochemistry, immucytochemistry and Western blot examines indicated positive results of ALP, OC, hBMP-2 after 48 hours.</p><p><b>CONCLUSION</b>Adipose tissue contains abundant ADSCs which could be transfected as gene vectors by adenovirus, ADSCs transfected with Ad-hBMP-2 can convert to ostoeblasts, and can act as a kind of seed cells for osteo-tissue engineering.</p>

Clinical study of oral and maxillofacial malignancies associated with multiple primary malignant neoplasms / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics, diagnosis and treatment strategy of oral and maxillofacial malignancies in multiple primary malignant neoplasms (MPMNs).</p><p><b>METHODS</b>21 cases of oral and maxillofacial malignancies associated with MPMNs admitted to our hospital between 1985 and 2000 were studied respectively.</p><p><b>RESULTS</b>There were 44 malignant cases in the 21 MPMNs patients. Among the 44 cases, there were 24 cases in alimentary and respiratory tract such as oral, pharynx, esophagus, stomach and lung, and 10 cases in salivary gland, breast and female reproductive system. There were 25 cases malignant neoplasms in oral and maxillofacial region where squamous cell carcinoma was the most common pathologic type, secondly adenoid cystic carcinoma. In oral and maxillofacial region, MPMNs were often found in tongue, parotid and submandibular gland, buccal mucosa and gingival.</p><p><b>CONCLUSION</b>Tongue and salivary gland were the common locations with MPMNs, and they were closely associated with alimentary and respiratory tract. Patients with malignancies of oral and maxillofacial region associated with MPMNs must be submitted to a long-term and careful follow-up. For female patients, breast and female reproductive system should be examined specially. Regular follow-up, early detection, early diagnosis, active and effective treatment can help to improve the survival quality of MPMNs patients.</p>

Repairing soft-tissue defection of ankle and foot by sural neurovascular pedicle fasciocutaneous reversed island flap / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the clinical application of sural neurovascular pedicle fasciocutaneous reversed island flap to repair soft-tissue defection of the ankle and foot.</p><p><b>METHODS</b>From Sep. 1994 to Oct. 2004,29 patients with soft-tissue defects in the ankle and foot were repaired by use of sural neurovascular fasciocutaneous reversed island flap, including 15 cases of traumatic defects, 11 cases of burns and 3 cases of chronic ulcer. The flap area ranged from 5 cm x 7 cm to 12 cm x 20 cm, and the length of pedicle from 5 cm to 12 cm.</p><p><b>RESULTS</b>The flaps survived totally in 27 cases, the distal necrosed partially and secondary free-skin grafting were further conducted in 2 cases. Twenty-one cases were followed-up for 3 to 60 months,the circulation, color and texture of the flaps were excellent and 2-point discrimination was 10 - 15 mm. The appearance and function of ankle joints were good.</p><p><b>CONCLUSION</b>This flap has sufficient blood supply and a high survival rate; It is convenient in design, dissection and without sacrifice of major arteries. So, it is an effective method for the reconstruction of soft-tissue defects in ankle and foot.</p>

Study of the expression of integrin alphaV beta3 in mandibular distraction osteogenesis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and distribution pattern of Integrin alphaV beta3 (ItgalphaV beta3) in course of distraction 44 adults New Zealand white osteogenesis and to quest for the function of ItgalphaV beta3 in distraction osteogenesis (DO).</p><p><b>METHODS</b>rabbits were selected and divided randomly into 3 groups: DO, bone fracture and normal group. There were 20, 20 and 4 rabbits in each group separately. Animal models were established and animals were killed at different time points. Sections were stained by IHC method. Distribution and expression pattern of ItgalphaV beta3 were observed by semi-quantitative technique, and the results were managed with statistic methods.</p><p><b>RESULTS</b>The expression of ItgalphaV beta3 was found both in the DO and bone fracture groups. The positive staining was seen mainly in vascular endothelial cells on cytomembrane and in cytoplasm. The staining intensity of group of DO was higher than that of the bone fracture group.</p><p><b>CONCLUSION</b>ItgalphaV beta3 plays an important role in promoting the process of DO.</p>

Mechanisms of p16 gene inactivation salivary adenoid cystic carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of p16 gene inactivation in salivary adenoid cystic carcinoma.</p><p><b>METHODS</b>53 cases of freshly excised salivary adenoid cystic carcinomas were studied. Polymerase chain reaction (PCR) and single-stranded conformation polymorphism analysis of polymerase chain reaction products (PCR-SSCP) were used to detect deletion and mutation of p16 gene in salivary adenoid cystic carcinomas. Methylation specific PCR (MSP) was used to detect the methylation status of p16 gene.</p><p><b>RESULTS</b>The homozygous deletion, mutation and hypermethylation of p16 gene were noted in 16 cases (30.2%), 4 cases (7.5%) and 26 cases (49.1%) respectively in 53 cases of salivary adenoid cystic carcinomas.</p><p><b>CONCLUSION</b>The main inactivation mechanisms of p16 gene in salivary adenoid cystic carcinoma were hypermethylation and homozygous deletion. The mutation p16 gene was rare in salivary adenoid cystic carcinoma.</p>

Surgical treatment of tonsillar cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the surgical treatment of tonsillar cancer.</p><p><b>METHODS</b>Twenty-four patients with tonsillar cancer were treated with surgery and postoperative radiotherapy. The choice of surgical procedure was decided on the condition of the lesion. The tumor was resected through the transoral approach, mandibular swing approach, mandibular resection approach or hyoid approach. Surgical defect was repaired by pectoralis major myocutaneous flap, sternohyoid myofascial flap, tongue flap or soft palate flap.</p><p><b>RESULTS</b>The 3- and 5-year survival rates were 76.0% and 60.8%. Function of chewing, deglutition, respiration and speech was restored well.</p><p><b>CONCLUSION</b>Method of total resection of the tonsillar carcinoma through the optimum approach is best chosen according to the condition of the lesion, while preserving the oropharyngeal function. When combined with postoperative radiotherapy, the survival rate and quality of life of patients can be improved.</p>