1.Mechanism of hsa-miRNA124-3p regulating the proliferation and migration of human lung cancer cell line NCI-H460
Ye WANG ; Wang XIE ; Jie ZHANG ; Jieying SHENG ; Zhongliang GUO
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(8):1051-1058
Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P<0.01), and KLF4 protein was lower in the cancer tissues than in the adjacent tissue (P<0.01). The bioinformatic analysis showed there is a theoretical binding site (5'-UGCCUUAA-3') of hsa-miRNA124-3p in 3'-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3'-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA12472 h after transfection (P<0.05 ). Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced (P<0.05) There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group (P>0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.
2.Diffusion tensor MR imaging in pediatric patients with periventricular leukomalacia
Sheng XIE ; Xuemei GUO ; Aiguo CUI ; Jiangxi XIAO ; Xuexiang JIANG
Chinese Journal of Radiology 2001;0(03):-
Objective To compare pediatric patients with periventricular leukomalacia (PVL) with normal children by using diffusion tensor MR imaging. Methods Diffusion tensor images were obtained in 15 pediatric patients with PVL and 15 age-matched normal children. Regions of interest were drawn to measure the fractional anisotropy (FA) in bilateral posterior limb of internal capsule, bilateral optic radiation, genu of corpus callosum, and splenium of corpus callosum. The values of PVL patients and normal children were compared using non-dependent samples T-test. Results The FA values of regions of interest prescribed were significantly lower in PVL patients than in normal children(P
3.Preliminary study of normal changes in brain white matter during childhood with diffusion tensor imaging
Jiangxi XIAO ; Xuemei GUO ; Sheng XIE ; Xiaoying WANG ; Xuexiang JIANG
Chinese Journal of Radiology 2000;0(12):-
Objective To study the normal changes in brain white matter during childhood by analyzing the anisotropy of different regions and different age groups with diffusion tensor imaging (DTI). Methods DTI was performed in 89 children (age range from 2 days to 18 years) without brain abnormalities, and the data measured in fractional anisotropy(FA) maps were analyzed statistically. Children less than 6 months were ranged to group 1, 6-12 months to group 2, 1-3 years to group 3, 3-5 years to group 4, 5-8 years to group 5, 8-12 years to group 6, 12-18 years to group 7. Results (1) There were significant differences in anisotropy (FA values) among different regions of white matter in brain In group 7, the FA value of corpus callosum was 0.826?0.039, middle cerebellar peduncle 0.678?0.043, frontal white matter 0.489?0.033. (2) The anisotropy among different age group was statistically different, P
4.Normal Changes in Brain During Childhood-Study of the Diffusion Tensor Imaging
Xuemei GUO ; Jiangxi XIAO ; Sheng XIE ; Xiaoying WANG ; Xuexiang JIANG
Journal of Practical Radiology 2001;0(10):-
Objective To study the normal changes in brain during childhood by analyzing the diffusion characters of different regions and different age phases with diffusion tensor imaging (DTI). Methods DTI was performed in 63 children (age range, 2 days to 9 years) without brain abnormalities and the data measured in ADC maps were analyzed statistically. Results (1) There were significant differences in ADC values among some different regions of brain tissue; (2) In babyhood, the ADC values in gray matters decreased faster than in white matters; (3) The ADC values decreased with the increasing of age , and show negatively exponentially correlation with age. Conclusion DTI shows the changes of brain tissue composition in vivo, with which normal changes in brain during childhood can be evaluated.
5.Primary Study of the White Matter in Normal Children with Color Tensor Map
Jiangxi XIAO ; Xuemei GUO ; Sheng XIE ; Xiaoying WANG ; Xuexiang JIANG
Journal of Practical Radiology 2000;0(12):-
Objective To study findings of the white matter of nomal children on color tensor map and evaluate the diffusion tensor imaging in the assessing development of the brain. Methods DTI was obtained in 89 normal children aging from 2 days to 18 years.Their color tensor maps were generated and analyzed.Results Their color tensor maps revealed different findings due to aging;with the increase of the age,the fiber tracts showed were more and well defined.Conclusion The color tensor map is superior to the routine MR in assessing the development of the white matter.
7.The prevention and treatment of heijingpaichitang for immune rejection following high-risk corneal transplantation in rats
Xiao-feng, XIE ; Qing-mei, TIAN ; Hong-sheng, BI ; Ying, WEN ; Shu, ZHANG ; Ai-lian, GUO
Chinese Journal of Experimental Ophthalmology 2012;(10):902-907
The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P<0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P<0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P>0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P<0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1% CsA.Heijingpaichitang and 0.1% CsA have a synergistic effect.
8.MRI findings in Leigh syndrome with SURF-1 gene 604G→C mutations
Li GUO ; Sheng XIE ; Jiangxi XIAO ; Zhaoyue QI ; Yanling YANG ; Xuexiang JIANG
Chinese Journal of Radiology 2008;42(12):1246-1248
Objective To determine the MR phenotype of Leigh syndrome (IS) with SURF-1 gene mutation.Methods The cranial MR examination of eight patients with the diagnosis of Leigh syndrome associated with SURF-1 gene 604G→C mutations were reviewed retrospectively.Comparison was made with typical LS patients' MRI.Observation was made regarding lesions in basal ganglia,subthalamic nuclei,substantia nigra,and dentate nuclei,involvement of white matter,and brain atrophy.Results The MR findings of LS in the study included the following:3 cases exhibited involvement of brain stem and subthalamic nuclei,2 of these 3 cases also had basal ganglia abnormalities.3 cases showed abnormality in the white matter without any involvement of the deep nuclei.Cerebral atrophy was observed in all cases in the group,and was the only finding in two cases.Conclusion The imaging findings in LS with SURF-1 604G→C gene mutation were variable.
9.Overexpression and RNAi vectors built for key secondary metabolic pathway genes PAL, HMGR, PGT of Arnebia euchroma.
Teng XIE ; Yu-Zhong LIU ; Sheng WANG ; Tan LIU ; Li-Ping KANG ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2014;39(23):4538-4543
Arnebia euchroma is the main source for medicinal herb Zicao. and its most important component shikonin compounds have high medicinal and industrial value. This research is aimed to build overexpression vectors and RNAi vectors for key secondary metabolism genes of A. euchroma, and bulid platform for constructions of related transgenic lines using GATEWAY technology. To build genetic material based genetic research platform is to provide a great convenience for digging and functional verification of the genes on secondary metabolic pathway, and also to fill the gaps in transgenic research of A. euchroma. This study is also important for the cultivation of shikonin high-yielding strains of A. euchroma.
Boraginaceae
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genetics
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metabolism
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Genetic Vectors
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genetics
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metabolism
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Plant Proteins
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genetics
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metabolism
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RNA Interference
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Secondary Metabolism
10.Transcriptome-based bioinformatics analysis of Arnebia euchroma ERF transcription factor family.
Teng XIE ; Sheng WANG ; Lei HUANG ; Xue WANG ; L-ping KANG ; Lan-ping GUO
China Journal of Chinese Materia Medica 2014;39(24):4732-4739
Twenty-seven ERF transcription factor family genes were isolated from Arnebia euchroma, with an average size of 1,010 bp, each gene encoded a 212 amino acids on average. The gene structure and expression of physicochemical properties, subcellular localization, signal peptides, senior structural domains and conservative forecasting, and analysis of A. euchroma were studied comparing with ERF gene gi261363612 of Lithospermum erythrorhizon, and phylogenetic analysis of A. euchroma ERF family was carried out. The results showed the existence of three conserved domains in this family, the senior structure based on random coil and it clustered into CBF/DREB and ERF subfamilies.
Amino Acid Motifs
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Amino Acid Sequence
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Boraginaceae
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genetics
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Cloning, Molecular
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Computational Biology
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Genome, Plant
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genetics
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High-Throughput Nucleotide Sequencing
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Medicine, Chinese Traditional
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Molecular Sequence Data
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Multigene Family
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Phylogeny
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Plant Proteins
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chemistry
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genetics
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Plants, Medicinal
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Sequence Analysis, DNA
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Transcription Factors
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chemistry
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genetics
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Transcriptome