ObjectiveTo systematically review the diagnostic value of FibroScan for the staging of liver fibrosis in chronic hepatitis B. MethodsWe searched the PubMed, EMbase, Web of Knowledge, CBM, WanFang Data and CNKI databases for studies investigated the diagnostic value of FibroScan for hepatic fibrosis B from Jan. 1st, 2003 to Aug. 31st, 2013. Two reviewers independently screened literature according to the exclusion and inclusion criteria, extracted data and assessed methodological quality of included studies. Then, Stata 13.0 software was used to analyze the data. ResultsA total of 15 studies involving 2 588 patients were included. The results of meta-analysis showed that:the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio and the AUC of SROC were 0.77 (95%CI 0.69 to 0.83), 0.84 (95%CI 0.70 to 0.87), 3.8 (95%CI 2.6 to 5.6), 0.29 (95%CI 0.22 to 0.38), 13 (95%CI 8 to 21), 0.82 (95%CI 0.82 to 0.88) for hepatic fibrosis; and were 0.81 (95%CI 0.73 to 0.87), 0.89 (95%CI 0.86 to 0.92), 7.5 (95%CI 5.3 to 10.3), 0.21 (95%CI 0.14 to 0.31), 36 (95%CI 20 to 65), 0.93 (95%CI 0.90 to 0.95) for early hepatic cirrhosis, respectively. ConclusionThe current evidence suggests that FibroScan is of good accuracy in the diagnosis of early hepatic fibrosis but not for hepatic cirrhosis in patient with chronic hepatitis B.
ObjectiveTo establish human bladder cancer cell line with silenced Fibulin-5 gene and observe the effects and mechanism of Fibulin-5 gene silencing on the proliferation activity and migration of the bladder cancer cells.MethodsThe human bladder cancer cells 5637 were divided into group F5 and group NC, and the cells in group F5 were infected with Fibulin-5 RNA interference (RNAi) lentivirus while the cells in group NC were infected with negative-control virus. Then the expression of Fibulin-5 mRNA was detected by real-time quantitative polymerase chain reaction, the cell proliferation activity was detected by MTT, the migration rate was detected by wound healing method, and the expression levels of proteins in receptor tyrosine kinase (RTK) pathway were detected by PathScan RTK Signaling Antibody Array Kit.ResultsThe Fibulin-5 mRNA expression decreased significantly by Fibulin-5 RNAi lentivirus (0.067±0.013 in group F5 vs. 1.001±0.000 in group NC), and the gene silencing efficiency reached 93.3%, so the Fibulin-5 silencing cell line was established successfully. Comparing with group NC, the relative absorbance value and migration rate of cell 5637 in group F5 decreased significantly (P<0.01); in addition, the expression levels of anaplastic lymphoma kinase, Axl, p44/42 mitogen activated protein kinase, and Src protein were up-regulated in group F5 (P<0.05).ConclusionFibulin-5 may play a role in the proliferation and migration of bladder cancer cells, and may have an inhibitory effect on extracellular signal-regulated kinase and its signaling pathway proteins.
OBJECTIVE: To build the trestle of tissue engineering for skin with the collagen. METHODS: The collagen was obtained from the baby cattle hide pretreated by Na2S and elastinase and Protease M, then the collagen was dissolved in 0.5 mol/L acetic acid solution. The collagen was treated with Protease N to minimize its immunogenicity. The resulting collagen could be used to build the trestle of tissue engineering for skin because of good biocompatibility. The collagen molecular weight and structure were analyzed by SDS-PAGE. The bioactivity of trestle was tested in the experiment of the mice wound healing and the cell implantation. RESULTS: The SDS-PAGE result of the collagen treated by Protease M showed the typical spectrum of type I collagen. The built trestle was a collagen sponge matrix in which micropore size was 50-200 microns. It could accelerate wound healing and the implanted fibroblasts could proliferate well. CONCLUSION: The collagen treated by Protease N can get good biocompatibilily and is suitable for building the trestles of tissue engineering for skin with good bioactivity.
Objective To summarize the function of fibula in stability of ankle joints.Methods Recent original articles were extensively reviewed, which were related to the physiological function and biomechanical properties of fibula, the influence of fibular fracture on stability of ankle joints and mechanism of osteoarthritis of ankle joints. Results The fibula had the function of weightbearing; and it was generally agreed that discontinued fibula could lead to intra articular disorder of ankle joint in children; but there were various viewpoints regarding the influence of fibular fracture on the ankle joint in adults. Conclusion Fibula may play an important role in stability of ankle joint.
In the study of repair of massive bone defect with free vascularized fibula graft, 13 cases were reported, in which traumatic defect in 7 cases, segmental resection of bone from tumors in 5 cases and osteomylitis in 1 cases. They all were treated successfully with vascularized fibular graft. After a follow-up of 6 months to 7 year, bone healing was observed with satisfactory and rehabilitation of functions. In one case, fatigued fracture occured twice due to early walking. It was concluded that free vascularized fibular graft was very helpful in the repair of massive bone defect, but prolonged external fixation after operation might be important to prevent fractur of grafted bone.
Objective To study the reparative and reconstructive for proximal humerus defect due to the excision of bone tumor with noninternal fixation non-vascularised fibular autografts. Methods From June 1991 toDecember 2003, 26 non-vascularised fibular grafts were used as substitutes for repair and reconstruction after resection for bone tumors on proximal humerus. Fifteen cases were given curettage and fibular supporting internal fixation, the other 11 cases were given tumor resection and joint reconstruction with proximal fibular graft. The age ranged from 6 to 41 years. Out of 26 patients, 5 had giant cell tumor, 9 had bone cysts, 8 had fibrous dysplasia and 4 had enchondroma. Results Twenty-six patients were followed up from 1 to 12 years (3.4 years on average). Local recurrence was found in 2 cases, and 1 of them died of lung metastasis. Both outlook and function of the reconstructed joints have good results in 15 proximal humeral joint surface reserved cases. Of them, 3 children gained normal shoulder function 3 weeks after operation. Part function were obtained in the other 11 fibular grafts substituted proximal humeral defect. Conclusion Non-vascularised fibular grafts is an appropriate treatment option for proximal humerus bone defect due to excision of bone tumor.
The treatmen t of the bone defect of the distal part of the radiu s included repair of the bone defect and resto rat ion of the funct ion of the w rist jo in t. Since 1979, th ree operat ive methods w ere u sed to t reat 13 cases, and they w ere graf t ing of the vascu larized f ibu la by anastom rsis f ibu lar vessels, graf t ing of upper part of f ibu lar w ith lateral inferio rgen icu lar artery and graf t ing vascu larized scapu la f lap. Follow up had been carried ou t from1 to 10 years. The resu lt w as sat isfacto ry. The discu ssion included the repair of the defect of the m iddle o r distal part of the radiu s, the operat ive methods, main at ten t ion s and indications. It was considered that it shou ld be based on the length of bone defect wh ile the operative method was considered.
Primary human hepatocytes (PHH) are the gold standard of in vitro human liver model for drug screening. However, a problem of culturing PHH in vitro is the rapid decline of cytochrome P450 (CYP450) activity, which plays an important role in drug metabolism. In this study, thermo-responsive culture dishes were used to explore the conditions for murine embryonic 3T3-J2 fibroblasts to form cell sheet. Based on the cell sheet engineering technology, a three-dimensional (3D) “sandwich” co-culture system of 3T3-J2 cell sheet/PHH/collagen gel was constructed. The tissue structure and protein expression of the model section were observed by hematoxylin eosin staining and immunofluorescence staining respectively. Phenacetin and bupropion were used as substrates to determine the activity of CYP450. The contents of albumin and urea in the system were determined by enzyme linked immunosorbent assay (ELISA). The results showed that the complete 3T3-J2 cell sheet could be obtained when the cell seeding density was 1.5×106 /dish (35 mm dish) and the incubation time at low temperature was 60 min. Through cell sheet stacking, a 3D in vitro liver model was developed. Compared with the two-dimensional (2D) model, in the 3D model, the cell-cell and cell-matrix connections were tighter, the activities of cytochrome P450 CYP1A2 and cytochrome P450 CYP2B6 were significantly increased, and the secretion levels of albumin and urea were increased. These indexes could be maintained stably for 21 d. Therefore, cell sheet stacking is helpful to improve the level of liver function of 3D liver model. This model is expected to be used to predict the metabolism of low-clearance drugs in preclinical, which is of great significance for drug evaluation and other studies.
OBJECTIVE: To analysis the biological characteristics of human fibroblasts transfected by human telomerase reverse transcriptase (hTERT) eucaryotic expression plasmid pGRN145. METHODS: Fibroblasts from children’s foreskin were isolated and cultured in vitro, and the fibroblasts were transfected by pGRN145 with Lipofec-tAMINE PLUS Reagent. After strict screening of hygromycin B, the positive clones were subcultured. The telomerase activity was detected by RT-PCR and TRAP-PCR technique. The cell generation cycle and apoptosis rate were detected by flow cytometry to investigate the proliferative characteristics after transfection, and the chromosome karyotype of transformed cells was analyzed. The collagen secreted by transformed cells was detected by immunohistochemical staining. RESULTS: The morphological properties of fibroblasts did not change obviously after transfection. There were telomerase activity in transfected fibroblasts, while it could not be detected in pre-transfection fibroblasts. The cell generation cycle had no obvious changes between pre-transfection and post-transfection. However, the apoptosis rate of transfected fibroblasts were decreased compared with that of pre-transfection. The fibroblasts transfected by pGRN145 maintained the normal diploid karyotype, as well as the cells could normally secret type I and III collagen. CONCLUSION: The human fibroblasts transfected by pGRN145 has telomerase activity with prolonged life span of culture, which preliminarily proves the availability of establishing standard seeding cell lines of tissue engineering by hTERT plasmid transfection techniques.
Objective To evaluate the clinical effects of fibula flap grafts on the repair of the extremities with traumatic compound tissue defects. Methods In 12 cases, the fibula flap grafts were employed to restore the extremities with traumatic compound tissue defects. Of the 12 patients, 9 were males, 3 were females; their ages ranged from 12 to 45. There were 2 cases of tibia defect combined with fibula fracture, 2 cases of tibia defect, 2 cases of radius defect, 3 cases of ulna defect, 1 case of calcaneus defect,and 2 cases of firstmetatarsus defect. The bone defect length ranged from 4.2 to 10.6 cm, 7.8 cm in average.The skin defect area ranged from 10.0 cm×4.5 cm to 27.0 cm×15.0 cm. The free transplantation of fibular flaps were used in 9 cases, the lapse operation were used in 2 cases, retrograde shift were used in 1 case. Results Postoperational vein crisis and commonperoneal nerve traction injury were observed in category mentioned above respectively. All the 12 fibula flaps survived after proper treatments such as removalof great saphenous vein. Follow-ups were done for 6 to 24 months. Both the transferred fibula and the recipient broken end reflected bones were healed. Four patients underwent the second-phase reconstruction operation oftendon moving power. One wrist and 1 ankle underwent arthrodesis in 3 to 6 months.All the effects were satisfactory. Conclusion The fibula flap grafts provide arelatively better alternative to repair the extremities with long bone compoundtissue defects. In addition, the sensory function reconstruction of fibula flaps should be given full attention.