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.
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.
Objective To evaluate the cytotoxicity of microdosis peracetic acid (PAA) so as to provide the evidence for making residual l imit of PAA steril ization. Methods Mouse fibroblasts (L929 cell l ine) cultured in vitro were observed to evaluate the influence of microdosis PAA including 1 × 10-6, 2 × 10-6, 3 × 10-6, 4 × 10-6, 5 × 10-6, and 10 × 10-6 (V/V). Theproliferation of cells was determined by MTT assay at 2, 4, and 7 days of culture. The growth curve and the relative growth rate (RGR) were obtained. The cytotoxicity of PAA at different concentrations was evaluated according to RGR. Results At 2, 4, and 7 days after culture, fibroblasts of 1 × 10-6 group grew with normal morphology analogous to control group, while the cell growth of other groups were poor. With the increase of PAA concentration, the absorbance (A) values decreased, which suggested that there was a significant negative correlation between cell prol iferation and PAA concentration. And the correlation coefficient was — 1.000 at 2 and 4 days, — 0.964 at 7 days. There was no significant difference in A value between 1 × 10-6 group and the control group (P gt; 0.05), while there were significant differences in A value between the control group and other concentration groups (P lt; 0.05). The growth curve of 1 × 10-6 group was similar to that of the control group, both had obvious phase of exponential growth. The growth curves of other groups had no obvious phase of exponential growth. The cytotoxicity of 1 × 10-6 group was classified as level 1, 2 × 10-6 group as level 2, 3 × 10-6 group as level 3, 4 × 10-6 group as level 3-4, 5 × 10-6 group and 10 × 10-6 group as level 4. Conclusion PAA of 1 × 10-6 had no obvious cytotoxicity. The residual l imit of PAA less than 1 × 10-6 was recommended.
Objective To evaluate the effect of the treatment of necrosis of femoral head with the free vascularized fibula grafting. Methods From October 2000 to February 2002, 31 hips in 26 patients with ischemic necrosis of the femoral head were treated with free vascularized fibula graft. Among these patients, 21 patients (25 hips) were followed up for 6-18 months(12 months on average). According to Steinberg stage:Ⅱ period, 5 hips;Ⅲ period,8 hips; Ⅳ period, 12 hips.Results Among 25hips, their Harris Hip Score at all satges were improved during the follow-up. The symptom of pain diminished or disappeared after operation. The patient’s ability to work and live was notlimited or only slightly limited during the follow-up. Radiographic evaluation showed that most femoral heads improved (18 hips) or unchanged (6 hips) and only oneworsened.Conclusion The free vascularized fibular grafting is a valuable method for femoral head necrosis. With this method, we can prevent or delay the process of the disease.
The material properties and volume proportion of the fibers as well as the cross-sectional area proportion of nucleus pulposus vary greatly in different studies. The effect of these factors on the mechanical behavior of intervertebral discs (IVDs) are uncertain. The IVDs finite element models with different parameters were created to investigate the pressure, height, rotation, stress, and strain of the IVDs under loads: pure compression, rotation after compression or axial moment after compression. The results showed that the material properties of fibers had great impact on the mechanical behavior of IVDs, especially on the rotation angle. When the fiber volume ratio was small, its changes had a significant impact on the rotation angle of the IVDs. The area proportions of nucleus pulposus had relatively little effect on the mechanical behavior of IVDs. The IVDs rotation should be observed when validating the model. By adjusting the elastic modulus or volume ratio of fibers within a reasonable range, a model that could simulate the mechanical behavior of normal IVDs could be obtained. It was reasonable to make the area proportion of nucleus pulposus within 25%–50% for the IVDs finite element model. This study provides guidance and reference for finite element modeling of the IVDs and the investigation of the IVDs degeneration mechanism.
OBJECTIVE In order to provide the scientific basis to find out a practical and effective method to evaluate the degree of muscle atrophy and a better method of prevention and treatment of skeletal muscle atrophy. METHODS Forty-two adult Spray-Dawley rats were used and the model of denervated gastrocnemius muscle was established by cutting off the tibial nerve. The muscle wet weight, diameter and cross section area of myocyte were measured. The motor end-plate, fibrillation potential amplitude and frequency of denervated skeletal muscle were observed. RESULTS The muscle wet weight rapidly reduced within 4 weeks. Afterwards, it maintained about 30 per cent of normal value, and the diameter and cross section area of myocyte progressively reduced. The motor end-plate slightly changed within 4 weeks, but its degeneration accelerated in 6 weeks and disappeared after 16 weeks. The fibrillation potential amplitude was maximum at 2 weeks and it progressively reduced after 12 weeks of muscle denervation. The changes of amplitude and frequency were consistent with the degeneration of end-plate. CONCLUSION The muscle wet weight, diameter and cross section area of myocyte, fibrillation potential amplitude and frequency could be considered as the morphological and electrophysiological indexes of muscle atrophy degree. It’s suggested that the repairing operation of peripheral nerve should be performed before the disappearance of motor end-plate.
Objective To observe the affection of optic nerve under acute ocular hypertension and the effect of protection of bFGF on optic nerve. Methods BSS was perfused into anterior chamber of rabbits to increase the intraocular pressure to cause retinal ischemia. A computer image analysis system was used to count the optic nerve axons.Eyes were intravitreally injected with bFGF and then the number of optic nerve axons of the normal rabbits,and hypertension with and without bFGE treatment groups were counted respectively. Results The number of optic nerve axons in ocular hypertension eyes was less than the normal eyes(P=0.00003).The bFGF treated eyes had more optic nerve axons than the controls(P=0.0078). Conclusions The acute ocular hypertension may cause the loss of the nerve axons,and bFGF may be effective in protecting optic nerve in acute ocular hypertension. (Chin J Ocul Fundus Dis,2000,16:94-96)
Objective To investigate the way to reconstruct bone scaffold afterremoval of giant benign bone tumor in extremities of children. Methods From June 1995 to October 2000, 6 cases of benign bone tumor were treated, aged 614 years. Of 6 cases, there were 4 cases of fibrous hyperplasia of bone, 1 case of aneurysmal bone cyst and 1 case of bone cyst; these tumors were located in humerus (2 cases), in radius (1 case), in femur (2 cases) and in tibia(1 case), respectively. All patients were given excision of subperiosteal affected bone fragment, autograft of subperiosteal free fibula(4-14 cm in length) and continuous suture of in situ periosteum; only in 2 cases, humerus was fixed with single Kirschner wire and external fixation of plaster. Results After followed up 18-78 months, all patients achieved bony union without tumor relapse. Fibula defect was repaired , and the function of ankle joint returned normal. ConclusionAutograft of subperiosteal free fibula is an optimal method to reconstruct bone scaffold after excision of giant benign bone tumor in extremities of children.
Objective To analyze MC3T3E1 cell morphology, prol iferation, and osteogenic differentiation in fibrin gel (FG) so as to lay a fundament for use of FG in tissue engneering. Methods MC3T3E1 cells were incubated in three concentrations (20, 10 and 5 mg/mL)of FG as the experimental groups (groups A, B and C) and in the common medium culture as the control group (group D). The cell morphology and distribution in FG were observed by inverted phase contrast microscope and confocal laser scanning microscope at different time. The cell prol iferation was assessed by fluorospectrophotometer. The alkal ine phosphatase (ALP) activity was detected by automatic biochemistry analyses and von Kossa staining was used to analyze calcium salts mineralization. RT-PCR was used to analyze the ALP and bone sialoprotein (BSP)mRNA expression at 14 and 21 days. Results In groups A, B and C, the MC3T3E1 cells had long processes which connected each other and formed network; but fusiform or cube cells were observed in group D at 21 days. The fluorescence intensity was increased gradually with time, was the highest at 14 days and the lowest at 28 days in group D; it was highest in groups A, B and C at 28 days, there were statistically significant differences when compared with group D (P lt; 0.05). The ALP activity was increased gradually with time, and it was the highest at 28 days in group D and at 21 days in groups A and B, there were significant differences (P lt; 0.05), no statistically significant differences compared with group D at other time points (P gt; 0.05). The mineral ization nodus were seen at 21 and 28 days in group A, but no mineral ization nodus was seen in group D at 28 days. The RT-PCR results showed the mRNA expressions of ALP and BSP were enhanced in group A when compared with group D (P lt; 0.05). Conclusion The osteogenic differentiation was most obvious and cell prol iferation was most active after 21 days of incubation in FG.
OBJECTIVE To investigate a good method for repairing the long bone defect of tibia combined with soft tissue defect. METHODS From 1988-1998, sixteen patients with long bone defect of tibia were admitted. There were 12 males, 4 females and aged from 16 to 45 years. The length of tibia defect ranged from 7 cm to 12 cm, the area of soft tissue defect ranged from 5 cm x 3 cm to 12 cm x 6 cm. Free fibula grafting was adopted in repairing. During operation, the two ends of fibular artery were anastomosised with the anterior tibial artery of the recipient, and the composited fibular flap were transplanted. RESULTS All grafted fibula unioned and the flap survived completely. Followed up for 6 to 111 months, 14 patients acquired the normal function while the other 2 patients received arthrodesis of the tibial-talus joint. In all the 16 patients, the unstable ankle joint could not be observed. CONCLUSION The modified method is characterized by the clear anatomy, the less blood loss and the reduced operation time. Meanwhile, the blood supply of the grafted fibula can be monitored.