A large number of data derived from molecular analyses support the hypothesis that human cancer is a genetic disease and a distinct subset of genes have been found to be genetically changed in most tumors. Molecular alterations in pancreatic cancer include: (1) oncogenes such as K-ras, c-myc, c-fos, and c-erbB-2; (2) tumor suppressor genes such as p53, p16, DPC4/SMAD4, and DCC; and (3) growth factors such as EGF, FGF, HGF, PDGF, VEGF, TGF-beta. Genetic alterations of K-ras and p53 are common in human pancreatic cancer, but the occurrence of pancreatic cancer is a multi-step phenomenon in which the accumulation of genetic changes is extremely important.
Epidermal Growth Factor ; genetics ; Fibroblast Growth Factors ; genetics ; Genes, Tumor Suppressor ; Genes, myc ; genetics ; Genes, p16 ; Genes, p53 ; genetics ; Genes, ras ; genetics ; Growth Substances ; genetics ; metabolism ; Humans ; Oncogenes ; genetics ; Pancreatic Neoplasms ; genetics

BACKGROUND: Congenital scoliosis (CS) is a complex spinal malformation of unknown etiology with abnormal bone metabolism. Fibroblast growth factor 23 (FGF23), secreted by osteoblasts and osteocytes, can inhibit bone formation and mineralization. This research aims to investigate the relationship between CS and FGF23. METHODS: We collected peripheral blood from two pairs of identical twins for methylation sequencing of the target region. FGF23 mRNA levels in the peripheral blood of CS patients and age-matched controls were measured. Receiver operator characteristic (ROC) curve analyses were conducted to evaluate the specificity and sensitivity of FGF23. The expression levels of FGF23 and its downstream factors fibroblast growth factor receptor 3 (FGFr3)/tissue non-specific alkaline phosphatase (TNAP)/osteopontin (OPN) in primary osteoblasts from CS patients (CS-Ob) and controls (CT-Ob) were detected. In addition, the osteogenic abilities of FGF23-knockdown or FGF23-overexpressing Ob were examined. RESULTS: DNA methylation of the FGF23 gene in CS patients was decreased compared to that of their identical twins, accompanied by increased mRNA levels. CS patients had increased peripheral blood FGF23 mRNA levels and decreased computed tomography (CT) values compared with controls. The FGF23 mRNA levels were negatively correlated with the CT value of the spine, and ROCs of FGF23 mRNA levels showed high sensitivity and specificity for CS. Additionally, significantly increased levels of FGF23, FGFr3, OPN, impaired osteogenic mineralization and lower TNAP levels were observed in CS-Ob. Moreover, FGF23 overexpression in CT-Ob increased FGFr3 and OPN levels and decreased TNAP levels, while FGF23 knockdown induced downregulation of FGFr3 and OPN but upregulation of TNAP in CS-Ob. Mineralization of CS-Ob was rescued after FGF23 knockdown. CONCLUSIONS Our results suggested increased peripheral blood FGF23 levels, decreased bone mineral density in CS patients, and a good predictive ability of CS by peripheral blood FGF23 levels. FGF23 may contribute to osteopenia in CS patients through FGFr3/TNAP / OPN pathway.
Humans ; Osteopontin/genetics* ; Alkaline Phosphatase/metabolism* ; Receptor, Fibroblast Growth Factor, Type 3/metabolism* ; Scoliosis/genetics* ; Osteoblasts/metabolism* ; Calcinosis ; RNA, Messenger/metabolism* ; Bone Diseases, Metabolic/metabolism* ; Fibroblast Growth Factors/genetics*

OBJECTIVETo study the instantaneous expression aFGF-GFP fusion gene in Carthamus tinctorius.

METHODMolecular biology methods were applied to construct aFGF and GFP fusion gene vector, it is transformed into C. tinctorius by Agrobacterium tumefaciens, forming the resistant callus, fluorescence microscopy was used for detection.

RESULTaFGF gene and GFP gene were amplified by PCR reaction. It was successfully constructed plant fluorescence expression vector pCAMBIA1390: :35S: :aFGF-GFP, it was used to transform C. tinctorius, and the acquired resistance calli showed strong green fluorescence under UV light.

CONCLUSIONThe expression of GFP in resistance C. tinrictorius calli is good, it is indicated that aFGF gene in plant cells has also been expressed.

Carthamus tinctorius ; genetics ; metabolism ; Cloning, Molecular ; Fibroblast Growth Factors ; genetics ; metabolism ; Gene Expression ; Green Fluorescent Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

DNA-Binding Proteins ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Receptor, Fibroblast Growth Factor, Type 1 ; genetics ; Transcription Factors ; genetics

The aim of the study is to construct two vectors for efficient expression of soluble recombinant proteins. The first vector was constructed by cloning the HisSUMO fragment into an expression vector pET30a(+) to fuse with the gene of interest (designated as HisSUMO Express). The second vector was constructed in the same way, but with a hydroxylamine cleavage site between HisSUMO and the gene of interest for an economic purpose (designated as HisSUMO Economic). The mouse fibroblast growth factor-21(mFGF-21), which was difficult to express in routine-used expression vectors, was taken as an example to test the vectors. The results showed that the mFGF-21 was expressed at high level in both vectors. The Sumo/mFGF-21 fusion protein accounted for more than 40% of the total bacterial protein. The fusion protein was purified with Ni-TNA column, and the HisSUMO was removed by cleavage of the fusion protein with either hydroxylamine solution or SUMO protease I. The concentration of the purified mFGF-21 mature protein was 54 mg/L and the recovery rate was 6%. The purified proteins derived from either hydroxylamine or SUMO protease I cleavage could stimulate glucose up-take by adipocytes. These results indicated that both HisSUMO Express and HisSUMO Economic were useful expression vectors for efficient expression of soluble recombinant proteins.
Animals ; Fibroblast Growth Factors ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Hydroxylamine ; chemistry ; Mice ; Peptide Hydrolases ; chemistry ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Solubility

OBJECTIVETo study the distribution of polymorphism of c.212-37insC (rs3832879) in intron 1 of fibroblast growth factor 23 (FGF23) gene and its association with Kawasaki disease (KD) and coronary artery lesions (CAL).

METHODSForty children with KD were enrolled in this study, among whom 16 children had concurrent CAL. Twenty-six age-matched healthy children were enrolled as controls. PCR and gene sequencing were applied to explore the distribution of polymorphism of c.212-37insC (rs3832879) in FGF23 gene in KD patients and controls.

RESULTSAmong 40 children with KD, 14 (35%) carried the polymorphism of c.212-37insC (rs3832879) in FGF23 gene; among 26 controls, 6 (23%) carried such polymorphism. There was no significant difference in genotype distribution at this locus between the two groups (P=0.30). Among 16 children with CAL, 9 (56%) carried the polymorphism at this locus; among 24 children without CAL, 5 (21%) carried such polymorphism. As for the comparison of two subgroups with and without CAL, the difference in genotype distribution at this locus had statistical significance (P=0.02, OR=4.89, 95% CI: 1.21-19.71).

CONCLUSIONSThe polymorphism of c.212-37insC (rs3832879) in FGF23 gene may not be associated with the pathogenesis of childhood KD, but it may be associated with the development of CAL in children with KD.

Child ; Child, Preschool ; Coronary Artery Disease ; etiology ; genetics ; Female ; Fibroblast Growth Factors ; genetics ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; etiology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo establish genetic transformation system of active fibroblast growth factor (aFGF) in Carthamus tinctorius.

METHODThe culture condition was optimized by orthogonal experiment design with cotyledon of C. tinctorius as the explant. The aFGF was transferred into safflower through Agrobacterium-mediated transformation and screened under different concentrations of antibiotics, and then PCR was identified.

RESULTIt confirmed the optimal differentiation medium: MS + BA 1.0 mg x L(-1) + NAA 0.2 mg x L(-1), the optimal root medium: 1/4 MS + NAA 2.0 mg x L(-1) + IAA0.5 mg x L(-1). The bacteriostatic effect of the three antibiotics showed slight difference. From them Tim was selected with the concentration of 400 mg x L(-1). It showed the bacteriostatic effect and promoted also differentiation. The selective concentration of hyg was confirmed to be 6 mg x L(-1). The eight transformed plants were identified, the positive rate was 25%.

CONCLUSIONIt was determined the best hormones and the ratios for the differentiation and rooting of the safflower by organogenesis. It was identified the optimal concentration of inhibitory antibiotics and selection antibiotics. The aFGF gene was cloned in a part of plant by PCR analysis. It is shown that the aFGF gene has been integrated into safflower genome.

Carthamus tinctorius ; genetics ; metabolism ; Fibroblast Growth Factors ; genetics ; metabolism ; Gene Expression ; Genetic Engineering ; methods ; Plants, Genetically Modified ; genetics ; metabolism ; Transformation, Genetic

The klotho gene was originally identified as a putative age-suppressing gene in mice that extends life span when overexpressed. It induces complex phenotypes resembling human premature aging syndromes when disrupted. The gene was named after a Greek goddess Klotho who spun the thread of life. Since then, various functional aspects of the klotho gene have been investigated, leading to the identification of multiple novel endocrine axes that regulate various metabolic processes and an unexpected link between mineral metabolism and aging. The purposes of this review were to overview recent progress on Klotho research and to discuss a novel aging mechanism.
Aging/genetics/*metabolism ; Animals ; Chronic Disease ; Fibroblast Growth Factors/metabolism ; Glucuronidase/genetics/*metabolism ; Homeostasis ; Humans ; Kidney Diseases/metabolism/physiopathology ; Phenotype ; Phosphates/metabolism ; Phosphorus, Dietary/metabolism ; *Signal Transduction

OBJECTIVETo investigate the expression sequence and distribution characteristics of the protooncogenes c-fos, c-myc and endogenous basic fibroblast growth factor (bFGF) genes in burned tissues, and to explore the possible effects of changes in these genes' functions on wound healing.

METHODSPartial-thickness burns of 30% TBSA were established on backs of Wistar rats. In situ hybridization and histological methods were used to detect expression of c-fos, c-myc and bFGF genes in normal and burned tissue at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d postburn.

RESULTSAlthough expression of c-fos and c-myc genes and bFGF gene could be found in normal skin, the expression of all three were markedly induced by burn wounds and the expression models in sequence and distribution were quite different. Expression of c-fos gene increased and peaked at 6 h. Signals were mainly localized in both nuclei of dermal fibroblasts and monocytes. The expression of bFGF gene increased at 6 h and peaked at 1 d postburn, and was distributed in the cytoplasm of fibroblasts. C-myc gene peaked 3 d postburn and was also distributed in the cytoplasm of fibroblasts.

CONCLUSIONSThese results indicated that thermal injury could induce the expression of c-fos, c-myc and bFGF at gene level, showing phasic control and regional distribution. The phasic expression of these genes suggests that there is an interaction between protooncogenes and bFGF, which may play an important role in wound healing. The different expressions of c-fos and c-myc play an inducing role in regulating bFGF, and in turn affect wound healing.

Animals ; Burns ; metabolism ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation ; Genes, fos ; Genes, myc ; In Situ Hybridization ; Male ; Rats ; Rats, Wistar ; Time Factors

Primary HLSCs were successfully cultured and assayed by AE5 in vitro. Constructed eukaryotic expressive vector of pEGFP-bFGF was transferred into the human limbal stem cells by the liposome-mediated technique, and 48 hours later, specific green fluorescence was observed by fluorescence microscope. The gene transfeetion efficiency was 20%-30%. Then the model of cells injury was created by use of NaOH. The cells were divided into four groups: Normal, bFGF, NaOH and bFGF+NaOH. The cellular viability in each group was measured by MTT colorimetry, and the cellular apoptosis rate and necrosis rate were observed by laser scanning confocal microscopy. The cellular viability in bFGF+NaOH group was higher than that in NaOH group (P < 0.05) ,while the cellular apoptosis rate plus necrosis rate displayed significant difference between the two groups (P < 0.05). The pEGFP-bbGF gene was noted to be successfully transferred into HLSCs and the cells were found growing well. These indicated that bFGF gene has a protective effect on the HLSCs injured by NaOH. We have also probed the feasibility of trying the treatment for ocular surface disease through gene engineering recombined tissue engineering.
Cells, Cultured ; Cornea ; cytology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Gene Expression ; Genetic Therapy ; methods ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cells ; cytology ; Tissue Engineering ; Transfection ; Vascular Endothelial Growth Factors ; genetics ; metabolism