A novel gain of function mutant in C-kit gene and its tumorigenesis in nude mice

AIM: To transfect mutant C-kit cDNA at codon 579 into human embryonic kidney cell line to observe its role in the pathogenesis of gastrointestinal stromal tumor (GIST).METHODS: Eukaryotic expression vectors of pcDNA3-Kit-NW and pcDNA3-Kit-W were constructed. Then pcDNA3-Kit-NW and pcDNA3-Kit-W plasmids were transfected into human embryonic kidney cell line by Lipofectamine. The resistant done was screened by G418filtration and identified by sequencing, Western blotting,and immunocytochemical staining. Human embryonic kidney cells were divided into three groups including pcDNA3-Kit-NW, pcDNA3-Kit-W, and vector control groups. Absorbency value with a wavelength of 574 nm was detected by MTT analysis. Mice were injected with three groups of cells. Volume, mass, and histological examinations of the tumors in different groups were measured and compared.RESULTS: The C-kit gene and mutant C-kit gene were successfully cloned into the eukaryotic expression vector pcDNA3. pcDNA3-Kit-NW and pcDNA3-Kit-W were successfully transfected into human embryonic kidney cell line and showed stable expression in this cell line.Cell proliferating activity had significant differences between pcDNA3-Kit-NW and pcDNA3, pcDNA3-KitNW and pcDNA3-Kit-W (P＜0.05), respectively. Tumors were only observed in nude mice implanted with cells transfected with pcDNA3-Kit-NW.CONCLUSION: Mutation of C-kit gene increases the proliferation activity of human cells and plays an important role in the malignant transformation of GIST.

Preparation of ferronickel from nickel laterite via coal-based reduction followed by magnetic separation

The sticking phenomenon between molten slag and refractory is one of the crucial problems when preparing ferronickel from laterite ore using rotary hearth fulnace or rotary kiln processes. This study aims to ameliorate sticking problems by using silicon dioxide （SiO2） to adjust the melting degree of file briquette during reduction roasting. Thermodynamic analysis indicates that the melting temperature of the slag gradually increases with an increase in the SiO2 proportion （SiO2/（SiO2 ＋ A1203 ＋ MgO） mass ratio）. Experimental validations also prove that the briquette retains its original shape when the SiO2 proportion is greater than 75wt%, and sticking problems axe avoided during reduction. A ferronickel product with 8.33wt% Ni and 84.71wt% Fe was prepared via reductive roasting at 1500~C for 90 min with a SiO2 proportion of 75wt% and a C/O molar ratio of 1.0 followed by dry magnetic separation; the corresponding recoveries of Ni and Fe reached 75.70% and 77.97%, respectively. The micro stxucture and phase txmlsformation of reduced briquette reveals that the aggregation and growth of ferroinckel particles were not significantly affected after adding SiO2 to the reduction process.

Angiopoietin-1 targeted RNA interference suppresses angiogenesis and tumor growth of esophageal cancer

AIM: To determine the inhibitory effect of the adenovirus- based angiopoietin-1 (Ang-1) targeted small interfering RNA expression system (Ad/Ang-1si) on the expression of the Ang-1 gene, cell growth and apoptosis in human esophageal cancer cell line Eca109. METHODS: siRNA-expressing adenovirus targeting Ang-1 gene was constructed using the Ad Easy System. Cultured Eca109 cells were transfected with Ad/Ang-1si (Eca109/Ang-1si), and Ad/si was used to infect Eca109 cells as control (Eca109/si). Ang-1 gene expression and concentration was determined with RT-PCR and ELISA, respectively. Human umbilical vein endothelial cell (HUVEC) migration and proliferation were analyzed. After s.c. injection into athymic nu/nu mice, the tumor growth, vessel density and apoptosis of each group was also determined. RESULTS: HUVEC migration induced by conditioned medium from Ang-1si-transfected Eca109 cells was significantly less than that induced by conditioned medium from Eca109 cells and control adenovirus- transfected Eca109 cells. Furthermore, after s.c. injection into athymic nu/nu mice, the tumor growth and cell apoptosis of Ad/Ang-1si -expressing Eca109 cells was significantly lower than that of parental or control adenovirus-transfected cells. Vessel density assessed by CD31 immunohistochemical analysis and Ang-1 expression by RT-PCR were also decreased. CONCLUSION: The targeting Ang-1 may provide a therapeutic option for esophageal cancer.

Isothermal reduction kinetics of Panzhihua ilmenite concentrate under 30vol% CO–70vol% N2 atmosphere

The reduction of ilmenite concentrate in 30vol% CO–70vol% N_2 atmosphere was characterized by thermogravimetric and differential thermogravimetric(TG–DTG) analysis methods at temperatures from 1073 to 1223 K.The isothermal reduction results show that the reduction process comprised two stages;the corresponding apparent activation energy was obtained by the iso-conversional and model-fitting methods.For the first stage,the effect of temperature on the conversion degree was not obvious,the phase boundary chemical reaction was the controlling step,with an apparent activation energy of 15.55–40.71 k J·mol^(–1).For the second stage,when the temperatures was greater than 1123 K,the reaction rate and the conversion degree increased sharply with increasing temperature,and random nucleation and subsequent growth were the controlling steps,with an apparent activation energy ranging from 182.33 to 195.95 k J·mol^(–1).For the whole reduction process,the average activation energy and pre-exponential factor were 98.94^(–1)18.33 k J·mol^(–1) and 1.820^(–1).816 min^(–1),respectively.

Effect of cooling rate on the crystallization behavior of perovskite in high titanium-bearing blast furnace slag

The effect of cooling rate on the crystallization of perovskite in high Ti-bearing blast furnace（BF） slag was studied using confocal scanning laser microscopy（CSLM）. Results showed that perovskite was the primary phase formed during the cooling of slag. On the slag surface, the growth of perovskite proceeded via the successive production of quasi-particles along straight lines, which further extended in certain directions. The morphology and structure of perovskite was found to vary as a function of cooling rate. At cooling rates of 10 and 30 K/min, the dendritic arms of perovskite crossed obliquely, while they were orthogonal at a cooling rate of 20 K/min and hexagonal at cooling rates of 40 and 50 K/min. These three crystal morphologies thus obtained at different cooling rates respectively corresponded to the orthorhombic, cubic and hexagonal crystal structures of perovskite. The observed change in the structure of perovskite could probably be attributed to the deficiency of O^2-, when Ti2O3 was involved in the formation of perovskite.

Stem cell factor-mediated wild-type KIT receptor activation is critical for gastrointestinal stromal tumor cell growth

AIM: To clarify the biological role of stem cell factor (SCF)-mediated wild-type KIT receptor activation in gastrointestinal stromal tumor (GIST) growth. METHODS: The co-expression of wild-type KIT receptor and SCF was evaluated in 51 GIST samples using mutation analysis and immunohistochemistry, and the results were correlated with clinicopathological param- eters, including the mitotic count, proliferative index (Ki-67 immunohistochemical staining), mitotic index (phospho-histone H3 immunohistochemical staining) and apoptotic index (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling). Using primary cultured GIST cells, the effect of SCF-mediated wild-type KIT receptor activation was determined by western blotting, methyl thiazolyl tetrazolium (MTT), and apoptosis assays. RESULTS: We found that wild-type KIT receptor and SCF protein were expressed in 100% and 76.5% of the 51 GIST samples, respectively, and the co-expression of wild-type KIT receptor and SCF was associated with known indicators of poor prognosis, including larger tumor size (P = 0.0118), higher mitotic count (P = 0.0058), higher proliferative index (P = 0.0012), higher mitotic index (P = 0.0282), lower apoptosis index (P = 0.0484), and increased National Institutes of Health risk level (P = 0.0012). We also found that the introduction of exogenous SCF potently increased KIT kinase activity, stimulated cell proliferation (P < 0.01) and inhibited apoptosis (P < 0.01) induced by serum starvation, while a KIT immunoblocking antibody suppressed proliferation (P = 0.01) and promoted apoptosis (P < 0.01) in cultured GIST cells. CONCLUSION: SCF-mediated wild-type KIT receptor activation plays an important role in GIST cell growth. The inhibition of SCF-mediated wild-type KIT receptor activation may prove to be particularly important for GIST therapy.

Isothermal reduction of titanomagnetite concentrates containing coal

The isothermal reduction of the Panzhihua titanomagnetite concentrates （PTC） briquette containing coal under argon atmosphere was investigated by thermogravimetry in an electric resistance furnace within the temperature range of 1250-1350°C. The samples reduced in argon at 1350°C for different time were examined by X-ray diffraction （XRD） analysis. Model-fitting and model-free methods were used to evaluate the apparent activation energy of the reduction reaction. It is found that the reduction rate is very fast at the early stage, and then, at a later stage, the reduction rate becomes slow and decreases gradually to the end of the reduction. It is also observed that the reduction of PTC by coal depends greatly on the temperature. At high temperatures, the reduction degree reaches high values faster and the final value achieved is higher than at low temperatures. The final phase composition of the reduced PTC-coal briquette consists in iron and fer-rous-pseudobrookite （FeTi2O5）, while Fe2.75Ti0.25O4, Fe2.5Ti0.5O4, Fe2.25Ti0.75O4, ilmenite （FeTiO3） and wustite （FeO） are intermediate products. The reaction rate is controlled by the phase boundary reaction for reduction degree less than 0.2 with an apparent activation energy of about 68 kJ·mol-1 and by three-dimensional diffusion for reduction degree greater than 0.75 with an apparent activation energy of about 134 kJ·mol-1. For the reduction degree in the range of 0.2-0.75, the reaction rate is under mixed control, and the activation energy increases with the increase of the reduction degree.

Numerical simulation of coupling multi-physical field in electrical arc furnace for smelting titanium slag

The smelting reduction process of the ilmenite in an electric arc furnace(EAF)is a commonly used technology for producing titanium slag in the world.It has particular significance to analyze the velocity-temperature-electromagnetics multi-physical field in an EAF for improving its productivity and reducing energy consumption.A transient three-dimensional mathematical model was developed to characterize the flow,heat transfer,and electromagnetic behavior in a titanium slag EAF.For describing the electromagnetic field and its effects on velocity and temperature distribution in the furnace,magnetohydrodynamic equations and conservation equations for mass,momentum,and energy were solved simultaneously by compiling the user-defined function program.The numerical model was verified by comparing with the literature data.The results indicate that the Lorentz force is the main driving force of the velocity and temperature distribution.Moreover,the influence of input current and location of electrodes on the multi-physical field distribution was also investigated.It is found that the appropriate range of input current and diameter of pitch circle are about 30,000 A and 3000-3500 mm,respectively.The mathematical model established can characterize the multi-physical field more accu-rately than before,which can provide valuable guidance for the operation improvement and design optimization of the EAF for producing titanium slag.

Formation behavior of CaTiO_3 during electrochemical deoxidation of ilmenite concentrate to prepare Fe–Ti alloy

The formation behavior of CaTiO3 during electro-deoxidization of ilmenite concentrate to prepare Fe–Ti alloy was investigated by experiments and simulation.The results indicate that the formation and decomposition of intermediate products,CaTiO3,are inevitable steps during electro-deoxidization of ilmenite concentrate.CaTiO3 can be generated through the hydrolyzation of molten salt and electrochemistry reaction during electrochemical process.The main reason for the generation of CaTiO3 is the electrochemistry reaction between Ca^2＋ from molten salt and TiO2 in the cathode.With the proceeding of the electro-deoxidization,CaTiO3 is further electrolyzed to form titanium sub-oxide.The current efficiency can be improved when CaTiO3 forms in the cathode by adding CaCO3 during sintering process.

Oxidation kinetics of ilmenite concentrate by non-isothermal thermogravimetric analysis

The non-isothermal oxidation experiments of ilmenite concentrate were carried out at various heating rates under air atmosphere by thermogravimetry.The oxidation kinetic model function and kinetic parameters of apparent activation energy（Ea）were evaluated by Málek and Starink methods.The results show that under air atmosphere,the oxidation process of ilmenite concentrate is composed of three stages,and the chemical reaction（G（α）=1-（1-α）~2,whereαis the conversion degree）plays an important role in the whole oxidation process.At the first stage（α=0.05-0.30）,the oxidation process is controlled gradually by secondary chemical reaction with increasing conversion degree.At the second stage（α=0.30-0.50）,the oxidation process is completely controlled by the secondary chemical reaction（G（α）=1-（1-α）~2）.At the third stage（α=0.50-0.95）,the secondary chemical reaction weakens gradually with increasing conversion degree,and the oxidation process is controlled gradually by a variety of functions;the kinetic equations are G（α）=（1-α）^（-1）（β=10K·min^（-1）,whereβis heating rate）,G（α）=（1-α）^（-1/2）（β=15-20K·min^（-1））,and G（α）=（1-α）^（-2）（β=25K·min^（-1））,respectively.For the whole oxidation process,the activation energies follow a parabolic law with increasing conversion degree,and the average activation energy is 160.56kJ·mol^（-1）.

Effect of pre-oxidation degree on gaseous reduction of pre-oxidized ilmenite concentrate by CO

The effect of pre-oxidation degree of Panzhihua ilmenite concentrate on the gaseous reduction by carbon monoxide was investigated.The raw ilmenite concentrates were pre-oxidized at 600,700,800,900,and 1000°C for 2 h,respectively.The phase change,FeO content,and morphology after the oxidation at various temperatures were characterized.The oxidation degree was calculated according to the change of FeO content.Thermogravimetry was performed on the oxidized ilmenite concentrate samples(oxidized at 600,800,and 1000°C)at 850,900,and 950°C,respectively.The results show that the pre-oxidation of ilmenite concentrate can accelerate the reduction process:the higher pre-oxidation degree,the faster reduction rate.The high pre-oxidation degree of ilmenite concentrate is beneficial to accelerate the subsequent reduction process in two ways.Three stages of the reduction process were divided via the first derivatives of reduction degree,and the reduction mechanism and apparent activation energy were discussed subsequently.