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Objective:To analyze the changes in the transcriptome of mouse fibroblasts after exposure to extremely low frequency electromagnetic fields (ELF-EMFs) using next-generation sequencing technology, and to screen out related pathways and genes that might be involved in the regulation of fibroblast growth by ELF-EMFs.Methods:The mouse NIH/3T3 cells were divided into the radiation group and the normal control group.The cells in the radiation group were placed in a 0.2 mT, 50 Hz electromagnetic radiation system, and the cells in the normal control group was placed in the same coil system under the same conditions without power.After 24-hour culture in a cell incubator, RNA was extracted.The next-generation high-throughput sequencing technology was used to sequence the transcriptomes of the two groups, and perform gene function annotation and signal pathway database analysis on the selected differential genes.Some highly expressed genes were screened out and verified by real-time fluorescent quantitative PCR.Results:A total of 17 980 genes were identified in the transcriptome sequencing, and there were 140 significantly differentially expressed genes (DEGs), of which 120 were up-regulated and 20 were down-regulated.DEGs were enriched in enzyme catalytic activity, cell metabolism process, biological regulation, biosynthesis and so on.According to the Kyoto Encyclopedia of Genes and Genomes analysis, the DEGs mainly involved 55 pathways, among which the most enriched 10 pathways were aminoacyl-tRNA biosynthesis, platelet activation, neurotrophin signaling pathway, renin-angiotensin system, etc., closely related to cell biosynthesis.The DEGs that might be involved in the post-irradiation stress of cells were further screened out, including mitogen activated protein kinase 12 ( MAPK12), neurotrophic tyrosine kinase receptor type 3 ( NTRK3), angiotensin Ⅱ receptor type 2 ( AGTR2), vascular endothelial growth factor ( VEGF), etc.Real-time fluorescent quantitative PCR results showed that the relative expression levels of MAPK12, NTRK3, AGTR2, VEGF mRNA in the radiation group were 2.389±0.003, 2.481±0.350, 2.354±0.081, 1.559±0.110, respectively, which were significantly higher than 1.011±0.190, 1.011±0.180, 1.007±0.150, 1.008±0.153, respectively in the normal control group ( t=12.540, 6.309, 13.710, 3.078; all at P<0.05). Conclusions:After the mouse fibroblasts were interfered with ELF-EMFs, the expression levels of MAPK12, NTRK3, AGTR2, VEGF and other genes are significantly up-regulated, which mainly involve neurotrophin signaling pathway, renin-angiotensin system and other pathways.These genes and pathways may be the main way that ELF-EMFs affect fibroblasts.
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Palladium hydrogel capped by β-cyclodextrins (Pdβ-CD) was prepared by a facile method with β-cyclodextrins and palladium(II) chloride,which were then modified onto the surface of gold electrode. The morphology and structure of the as-prepared palladium hydrogel were characterized by X-ray powder diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), while the electrochemistry behaviors of gold electrode modified by Pdβ-CDwere investigated by cyclic voltammetry (CV) and differential pulse voltammetry(DPV). The results indicated the sensor had high electrochemistry response to hydrazine hydrate in the presence of K+,Na+,Mg2+,NH+4,Ni+2,Mn2+,Cl-,NO-3,SO2-4,PO3-4,HCOO-, C6H5O-3. Under the optimized conditions, the oxidized peak current showed linear relationship with the concentration of hydrazine hydrate in the concentration range of 25-950 μmol/L and the limit of detection (LOD) of 1.6 μmol/L(S/N=3).Owing to the facile preparation,high sensitivity and selectivity,the sensor has potential applications in determination of hydrazine hydrate in real water samples
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BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.
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BACKGROUND: Tissue engineering has a promising prospect in corneal transplantation. Scaffolds are always restricting the development of tissue-engineered cornea. OBJECTIVE: To analyze application of different seed cells and scaffolds, and to summarize the progress of tissue-engineered cornea in recent years.METHODS: First author searched literature from CNKI (2000/2010-10) and PubMed database (2000/2010-10). The key words are tissue engineering, corneal transplantation in Chinese or English. A total of 223 literatures were seized by computers, according to the inclusion criteria, papers concerning the advance, application and reconstruction of tissue-engineered cornea were analyzed. Finally, 33 papers were included for further analysis. The present study was to analyze the seed cells, scaffolds, organ building and clinical applications of tissue-engineered cornea and investigate the development direction in the future. RESULTS AND CONCLUSION: The results show that seed cells and scaffolds are the focus of the studies now. Corneal transplantation has a high success rate in organ transplantation since the immune privilege of eye and avascular cornea, and it will be able to be a tissue engineering organ that can be largely built, easy to transplant. Reconstruction of cornea has reached first base, but every kind of scaffold has certain drawbacks, so the next goal is to find an ideal scaffold material.
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The dominant gene Xa21 with broad-spectrum and high resistance to Xanthomonas oryzae pv. oryzae (Xoo) was transferred into C418, an important restorer line of japonica hybrid rice in China using double right-border (DRB) T-DNA binary vector through Agrobacterium-mediated transformation. 17 transgenic lines were Xa21-positive with high resistance to the race P6 of Xoo through PCR analysis and resistance identification, among the total 27 independent primary transformants (T0) obtained. The subsequent analysis of the T1 progenies of these 17 T0 lines through PCR-assisted selection and resistance investigation showed that four Xa21 transgenic T0 lines could produce selectable marker-free (SMF) progenies. The frequency of primary transformants producing SMF progenies was 15%. In addition, PCR analysis also revealed these SMF progenies did not contain vector backbone sequence, and they were named as SMF and vector backbone sequence-free (SMF-VBSF) Xa21 transgenic plants. The further molecular and phenotypic analysis of the T2 and T3 progenies testified the homozygous SMF-VBSF Xa21 transgenic plants were obtained with high resistance to Xoo.
Subject(s)
DNA, Bacterial , Genetics , Genetic Vectors , Oryza , Genetics , Plant Proteins , Genetics , Plants, Genetically Modified , Genetics , Protein Serine-Threonine Kinases , Genetics , Rhizobium , Genetics , Transformation, Genetic , XanthomonasABSTRACT
G46B is a promising holding line used for three-lines breeding strategy in hybrid rice, but it is susceptible to blast disease caused by Pyricularia grisea. To improve its blast resistance, three rice varieties, Digu, BL-1, and Pi-4, with blast resistance genes, Pi-d(t), Pi-b, and Pi-ta2, respectively, were used to be crossed with G46B, and 15 plants with these three blast resistance genes, Pi-d(t)1, Pi-b, and Pi-ta2, were selected from their F2 and B1C1 populations via a marker-aided crossing procedure. Among them, four plants were heterozygotes in the three resistance genes, with the genotype of Pi-d(t)1 pi-d(t)/Pi-b pi-b/ Pi-ta2 pi-ta2; ten plants were heterozygotes in two of the three resistance genes, of which six with the genotype of Pi-d(t)1 Pi-d(t)1/Pi-b pi-b/Pi-ta2 pi-ta2, three with the genotype of Pi-d(t)1 pi-d(t)1/Pi-b pi-b/Pi-ta2 Pi-ta2, and one with the genotype of Pi-d(t)1pi-d(t)1/Pi-b Pi-b/Pi-ta2 pi-ta2; and only one plant was homozygote in two of the three resistance genes with the genotype of Pi-d(t)1 Pi-d(t)/Pi-b pi-b/Pi-ta2 Pi-ta2. These results demonstrate the capacity of maker-assisted selection (MAS) in gene pyramiding for rice blast resistance and its enhancement for the efficiency in rice resistance breeding.
Subject(s)
Crosses, Genetic , Genes, Plant , Genetic Markers , Genotype , Oryza , Genetics , Plant Diseases , Genetics , Selection, GeneticABSTRACT
By using rice SSRP, RAPD and AFLP molecular markers, the genome of rice transgenic line "Minghui 63-Xa21" was analyzed. 32 SSRP primers, 42 RAPD primers and 8 AFLP primers could produce obvious PCR bands in the analysis of at least 12 individual plants selected randomly from "Minghui 63-Xa21" T3 generation. Totally 550 PCR bands, equivalent to 550 genomic sites, were detected. Different individual plants of the transgenic homozygous line displayed almost the same PCR pattern. Compared with the control "Minghui 63", no difference was found in their PCR patterns. This indicated that the introduction of Xa21 into the genome of "Minghui 63" did not change these 550 genome sites and their heredity. Very few variant PCR bands were observed in some individual plants from both "Minghui 63-Xa21" and "Minghui 63". However, the variant percentage was equivalent between the transgenic line and the non-transgenic control line.
Subject(s)
Chromosome Mapping , Methods , Genome, Plant , Microsatellite Repeats , Genetics , Oryza , Genetics , Plant Proteins , Genetics , Plants, Genetically Modified , Protein Serine-Threonine Kinases , Genetics , Random Amplified Polymorphic DNA Technique , MethodsABSTRACT
A large series of lens were collected from normal and senile cataractous human eyes, and were investigated according to the theories of oxyradical and lipid peroxide (LPO). The findings showed that the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase in the senile cataractous lens were very significantly lower than those in the normal ones, while the content of malondialdehyde (MDA) in the lens of senile cataractous eyes was very significantly higher than that of normal eyes. The results suggest that the oxyradical and LPO might be one of the direct causes leading to senile cataract formation.