OBJECTIVE To established a viable and convenient method for human nasoseptal chondrocyte culture in vitro and study the biological characters of the chondrocyte. METHODS The chondrocytes were obtained by incomplete enzyme digestion of cartilage and were seeded with small piece of cartilage in 25 ml culture flasks coated with polylysine previously. This study determines the biologic synthesis of the chondrocytes by toluidine blue and type Ⅱcollagen immunohistochemistry reaction. The proteoglycan synthesis efficiencies of human nasoseptal chondrocytes cultured in vitro were measured by using 35S-Na2SO4 incorporation. The modified method and the conventional method in proteoglycan synthesis were also compared. RESULTS The results suggested that human nasoseptal chondrocytes cultured by modified method still show the normal structure and morphology, and maintain the function of metabolism and self duplication. With the modified method, we can get a higher cell survival rate and more cells. CONCLUSION The modified method is a viable and convenient method.

Graft Rejection ; Humans ; Liver Transplantation

0.05).However,the shorter the history of the disease,the better the efficacy of the treatment.The younger the patient was,the better the efficacy of the treatment.Conclusions Vulva dystrophy can be treated with focused ultrasound effectively and safely.This approach appears to be a new promising treatment method.

OBJECTIVETo study the effect of melatonin on proliferation and apoptosis of fibroblasts in human hypertrophic scar and its mechanism.

METHODSFibroblasts from human hypertrophic scar were isolated and cultured with DMEM medium containing 10% FBS, and then they were divided into control (C, added with ethanol), low concentration (LC, added with 1 × 10(-5) mmol/L melatonin), middle concentration (MC, added with 1 × 10(-3) mmol/L melatonin), and high concentration (HC, added with 1 mmol/L melatonin) groups according to the random number table. After being cultured for 24 hours, cell morphologic change was observed under microscope; XTT-PMS assay was used to examine cell proliferative activity; cell cycle and apoptosis were assessed with flow cytometry after double staining of FITC and PI, and the levels of cyclin E, p53, and Fas mRNA were determined with fluorescence quantitative RT-PCR. Data were processed with analysis of variance and LSD test.

RESULTS(1) Fibroblasts in C group were spindle-shaped with growth in colonies. Along with the increase in melatonin concentration, fibroblasts in LC, MC, and HC groups gradually dispersed, deformed and atrophied, with shrunk cellular membrane, and decrease in ratio of nucleus and cytoplasm. (2) Proliferative activity of fibroblasts in LC, MC, and HC groups decreased along with an increase in melatonin concentration (1.49 ± 0.15, 1.24 ± 0.20, and 0.92 ± 0.09), which were lower that in C group (1.79 ± 0.10, F = 67.61, P < 0.05). Cell ratios of S and G2/M phases in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were all lower than those in C group [(10.6 ± 1.1)%, (6.1 ± 1.2)%, (3.2 ± 0.8)% vs.(16.9 ± 1.3)%, F = 286.10, P < 0.05; (13.5 ± 1.1)%, (9.8 ± 1.0)%, (6.0 ± 0.7)% vs. (16.7 ± 1.6)%, F = 162.69, P < 0.05]. Apoptotic rates in early and late stages of LC, MC, and HC groups increased along with an increase in melatonin concentration, all higher than those in C group (with F value respectively 424.05, 236.44, P values all below 0.05). The expressions of cyclin E mRNA in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were lower than that in C group (1.58 ± 0.21, 0.90 ± 0.20, and 0.24 ± 0.12 vs. 2.90 ± 0.30, F = 266.79, P < 0.05), while the expressions of p53 mRNA and Fas mRNA showed opposite tendency (with F value respectively 10.11, 12.03, P values all below 0.05).

CONCLUSIONSMelatonin can inhibit proliferation and induce apoptosis of fibroblasts in hypertrophic scar through regulating the gene expressions of cyclin E, p53, and Fas.

Adult ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin E ; metabolism ; Female ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Male ; Melatonin ; pharmacology ; Oncogene Proteins ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo investigate whether there is a association between estrogen receptor beta (ERb) gene polymorphism and primary hepatocellular carcinoma (HCC).

METHODS100 primary HCC patients and 100 controls from southwestern China were recruited in this study. The polymorphism of RsaI and AluI in ERb gene was analyzed by PCR- restriction fragment length polymorphism (RFLP).

RESULTSR allelic frequency was 35.0% and 51.0% in HCC patients and in control group, respectively, odds ratio (OR) was 0.517 [95% confidence intervals (CI) = 0.346-0.773], P less than 0.01. A allelic frequency was 20.5% and 11.0% in HCC patients and in control group, respectively, OR was 2.086 (95% CI = 1.191-3.654), P less than 0.01. Gene frequency of RsaI and AluI in the two groups was distributed with polymorphism.

CONCLUSIONERb gene polymorphism is associated with primary liver cancer. R allele may be the guard factor, and A allele may be its risk factor.

Carcinoma, Hepatocellular ; genetics ; Estrogen Receptor beta ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Polymorphism, Genetic

OBJECTIVETo study the inhibition effect of celastrol on neovascularization.

METHODSThe effect of celastrol on the in vitro proliferation of endothelial cell of vessel (ECV) was examined by MTT assay. The effect of celastrol on endothelial cell migration, tube formation on Matrigel and Chick chorioallantoic membrane angiogenesis was also examined. Matrigel plug assay was used to evaluate the effect of celastrol on angiogenesis in vivo.

RESULTSThe proliferation of ECV was inhibited significantly by celastrol with IC(50) being 1.33 microg/ml. Celastrol inhibited endothelial cell migration and tube formation in a dose-dependent manner. Celastrol also inhibited angiogenesis both in Matrigel plug of mouse model and in chick chorioallantoic membranes.

CONCLUSIONCelastrol, which can inhibit angiogenesis, could be developed as an antiangiogenic drug.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Endothelial Cells ; drug effects ; Mice ; Mice, Inbred BALB C ; Triterpenes ; pharmacology

BACKGROUNDIt is generally accepted that gliomas are the most common primary brain tumors with poor prognosis. We aimed to explore the relationship of the immunity of the central nervous system and the genesis and development of glioma.

METHODSG422 glioma was implanted in the brain of BALB/c mice (immuno-competent mice), nude mice (T cell related immuno-deficient) and complement C3 knock-out mice (complement C3 related immunodeficient). The survival time of the host, growth and histopathology of the tumor, and concentrations of tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ) in tumor tissues were assessed.

RESULTSTumor spheres were formed in all mice after injection, and glial fibrillary acidic protein (GFAP) positive staining of the cells declared their glioma origin. The longest median survival time of (44.3 ± 6.0) days was found in BALB/c mice, followed by (24.8 ± 5.2) days in nude mice and the shortest (18.6 ± 5.8) days in complement C3 knock-out mice. Accordingly, the growth of the tumor was fastest in complement C3 knock-out mice, followed by the nude mice and slowest in the BALB/c mice. Although the proportions of infiltrating CD68(+) lymphocytes in tumor tissues showed no significant difference (P > 0.05), TNF-α level in the nude and C3 knock-out mice, (28.11 ± 4.86) µmol/L and (22.87 ± 6.36) µmol/L respectively, were significantly lower (P < 0.01) than that in the BALB/c mice, which was (230.21 ± 39.17) µmol/L. The INF-γ level was highest in the BALB/c mice ((180.76 ± 29.19) µmol/L), followed by the nude mice ((113.46 ± 23.76) µmol/L) and then the C3 knock-out mice ((16.84 ± 4.45) µmol/L).

CONCLUSIONSThe G422 glioma implanted in the brains of mice with different immune ability would be a useful model for studying the relationship of the immune system and tumor in the central nervous system. Furthermore, the T cells and complement C3 compartments of the immune response may affect the growth of implanted tumors and inflammatory factors such as TNF-α and INF-γ.

Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; Complement C3 ; genetics ; metabolism ; Glioma ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Mice, Nude ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To investigate the potential value of 18F-fluorodeoxyglucose (FDG) PET/CT in predicting the survival of patients with primary tracheal malignant tumors. Materials and Methods: An analysis of FDG PET/CT findings in 37 primary tracheal malignant tumor patients with a median follow-up period of 43.2 months (range, 10.8–143.2 months) was performed. Cox proportional hazards regression analyses were used to assess the associations between quantitative 18F-FDG PET/CT parameters, other clinic-pathological factors, and overall survival (OS). A risk prognosis model was established according to the independent prognostic factors identified on multivariate analysis. A survival curve determined by the Kaplan-Meier method was used to assess whether the prognosis prediction model could effectively stratify patients with different risks factors. Results: The median survival time of the 37 patients with tracheal tumors was 38.0 months, with a 95% confidence interval of 10.8 to 65.2 months. The 3-year, 5-year and 10-year survival rate were 54.1%, 43.2%, and 16.2%, respectively. The metabolic tumor volume (MTV), total lesion glycolysis (TLG), maximum standardized uptake value, age, pathological type, extension categories, and lymph node stage were included in multivariate analyses. Multivariate analysis showed MTV (p = 0.011), TLG (p = 0.020), pathological type (p = 0.037), and extension categories (p = 0.038) were independent prognostic factors for OS. Additionally, assessment of the survival curve using the Kaplan-Meier method showed that our prognosis prediction model can effectively stratify patients with different risks factors (p < 0.001). Conclusion This study shows that 18F-FDG PET/CT can predict the survival of patients with primary tracheal malignant tumors. Patients with an MTV > 5.19, a TLG > 16.94 on PET/CT scans, squamous cell carcinoma, and non-E1 were more likely to have a reduced OS.

OBJECTIVETo investigate angiogenesis of collagen-chitosan porous scaffold, and to study survive of skin grafts on the scaffold after bilayer dermal equivalent (BDE) was transplanted on wounds with full thickness skin defects.

METHODSThe full thickness skin defects were made on 10 Bama miniature pigs and the BDE composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wound. Angiogenesis in dermal equivalent, wound healing, and healing and survive of skin grafts on dermal equivalent were observed in 1, 2, and 3 weeks after the BDE transplantation. At the same time, CD34 positive signals (neo-forming micro-vessels) were detected by immunohistochemical staining.

RESULTSInflammatory cells and fibroblasts infiltrated into dermal equivalent and a few new micro-vessels had been formed in 1 week after the BDE transplantation; neo-forming micro-vessels perpendicular to wound bed had increased significantly in 2 weeks after the BDE transplantation; neo-forming micro-vessels could be observed in almost all dermal equivalents in 3 weeks after the BDE transplantation. CD34 positive signals (neo-forming micro-vessels) in 3 weeks after the BDE transplantation was much more than those in 2 weeks after the BDE transplantation, and CD34 positive signals in 2 weeks after the BDE transplantation was much more than those in 1 week after the BDE transplantation. Survival rate of intermediate split thickness skin graft were 10%, 70% and 100% respectively after the skin grafts had been grafted for 2 weeks on surface of the scaffold which had been transplanted for 1, 2 and 3 weeks. Epidermis which had been grafted on surface of the scaffold for 1 or 2 weeks could perfectly survive after BDE had been transplanted for 1 or 2 weeks.

CONCLUSIONSCollagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect and can induce fibroblast infiltration and new micro-vessel formation. Epidermis grafted on surface of collagen-chitosan porous scaffold can perfectly repair wounds, and it has brilliant applied prospects in repairing skin defect.

Animals ; Chitosan ; Collagen ; Disease Models, Animal ; Female ; Graft Survival ; Neovascularization, Physiologic ; Silicones ; Skin ; injuries ; Skin Transplantation ; Swine ; Swine, Miniature ; Tissue Scaffolds ; Wound Healing

BACKGROUNDCatheter-directed thrombolysis (CDT) has been a mainstay in treating deep venous thrombosis (DVT). However, the optimal dosage of a thrombolytic agent is still controversial. The goal of this study was to evaluate the safety and efficacy of low dosage urokinase with CDT for DVT.

METHODSA retrospective analysis was performed using data from a total of 427 patients with DVT treated with CDT in our single center between July 2009 and December 2012. Early efficacy of thrombolysis was assessed with a thrombus score based on daily venography. The therapeutic safety was evaluated by adverse events. A venography or duplex ultrasound was performed to assess the outcome at 6 months, 1 year and 2 years postoperatively.

RESULTSThe mean total dose of 3.34 (standard deviation [SD] 1.38) million units of urokinase was administered during a mean of 5.18 (SD 2.28) days. Prior to discharge, Grade III (complete lysis) was achieved in 154 (36%) patients; Grade II (50-99% lysis) in 222 (52%); and Grade I (50% lysis) in 51 (12%). The major complications included one intracranial hemorrhage, one hematochezia, five gross hematuria, and one pulmonary embolism. Moreover, no death occurred in the study.

CONCLUSIONSTreatment of low-dose catheter-directed thrombosis is an efficacious and safe therapeutic approach in patients with DVT offering good long-term outcomes and minimal complications.

Adolescent ; Adult ; Aged ; Drug Administration Schedule ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Urokinase-Type Plasminogen Activator ; administration & dosage ; adverse effects ; therapeutic use ; Venous Thrombosis ; drug therapy ; Young Adult