1.Laryngeal obstruction caused by giant thyroglossal duct cyst: one case report.
Xiangbo SUN ; Jing YI ; Jian LV
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(4):370-371
Thyroglossal duct cyst is the most common congenital malformation of the neck. It is generally with non-specific symptoms. In our hospital, one case of laryngeal obstruction caused by giant thyroglossal duct cyst was cured, and this case was reported for reference.
Airway Obstruction
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Humans
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Laryngeal Diseases
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Thyroglossal Cyst
2.Mechanism of emodin enhancing chemosensitivity of pancreatic cancer cells
Wei WANG ; Yueping SUN ; Jing YI ; Jian WANG
Chinese Journal of Digestive Surgery 2010;09(5):353-356
Objective To investigate whether emodin enhances chemosensitivity of pancreatic cancer cells and the mechanism. Methods Normal human skin fibroblasts and pancreatic BXPC-3 cells were divided into control group, cisplatin (5 mg/L) treatment group, gemcitabine (0.5 μmol/L) treatment group, emodin (50 μmol/L)+ cisplatin (5 mg/L) co-treatment group and emodin (50 μmol/L) + gemcitabine (0.5 μmol/L) co-treatmentgroup. The cell viability was detected by MTT assay, the cell apoptosis rate by flow cytometry, the expression of the multidrug resistance-associated protein 1 ( MRP1 ), MRP2, breast cancer resistance protein (BCRP) and P-glycoprotein (P-gp) mRNA in different cell lines was determined by reverse transcription polymerase chain reaction.All data were analyzed using the t test. Results The cell viability and apoptosis rate of pancreatic BXPC-3 cells after treatment were 62.44% ± 3.42% and 27. 10% ± 4.24% in the cisplatin treatment group, and they were 30.53% ±0.05% and 66.33% ±9.37% in the emodin + cisplatin co-treatment group. There were significant differences in cell viability and apoptosis rate between the two groups (t = 13.20, 5. 35, P < 0.05 ). The cell viability and apoptosis rate of pancreatic BXPC-3 cells after treatment were 79.82% ±2.83% and 13.48% ± 1.65%in the gemcitabine treatment group, and they were 45.65% ± 2.46% and 62.74% ± 10. 18% in the emodin +gemcitabine co-treatment group. There were significant differences in cell viability and apoptosis rate between the two groups (t = 12.89, 8.28, P < 0. 05 ). There was no significant difference in the viability of normal human skin fibroblasts between the cisplatin treatment group and the emodin + cisplatin co-treatment group, and also between the gemcitabine treatment group and the emodin + gemcitabine co-treatment group (t = 2. 08, 0. 64, P >0.05 ). Expression of MRP1 mRNA was detected in pancreatic BXPC-3 cells, whereas the expression of MRP2,BCRP and P-gp mRNA was undetectable. The expression of MRP1 mRNA in pancreatic BXPC-3 cells was significantly down-regulated in the cisplatin treatment group, and cisplatin + emodin co-treatment had an additive effect on down-regulating the expression of MRP1 mRNA. Conclusion Emodin may enhance chemosensitivity of pancreatic cancer cells to cisplatin by down-regulating the expression of MRP1 mRNA.
4.Identification of chloride channel accessory 1 as a protective factor for the prognosis of colon cancer by weighted gene co-expression network and differential gene expression analysis
Zexin ZHANG ; Wenfeng WU ; Jing LI ; Xiaolan JIAN ; Yi YU
Chinese Journal of Digestion 2021;41(5):336-343
Objective:To screen the differentially co-expressed genes in the mRNA expression profile of colon cancer by combined application of weighted gene co-expression network analysis(WGCNA) and differential gene expression analysis, and to analyze the relationship between differentially co-expressed genes and prognosis.Methods:The transcriptomics data of the cancer genome atlas (TCGA)-colon adenocarcinoma (COAD) dataset and chip expression profile data of GSE68468 dataset were downloaded from TCGA and gene expression omnibus (GEO) databases based on bioinformatics methods, and differentially expressed gene (DEG) and the most significantly related weighted gene modules between normal tissues and colon cancer tissues were screened. Then, the differentially co-expressed genes related to colon cancer were screened out according to the intersection of differential genes and weighted genes. A protein-protein interaction (PPI) network was constructed, and the top ten core differentially co-expressed genes according to the maximal clique centrality (MCC) score were screened out by MCC calculation method. The expression of core genes in normal tissues and colon cancer tissues were further verified by TCGA-COAD dataset. Kaplan-Meier survival analysis was used to investigate the correlation between core genes and overall survival time and disease-free survival time of patients. The survival-related differentially co-expressed genes were verified by immunohistochemical staining in human protein atlas (HPA) database.Results:A total of 3 481 DEG of the TCGA-COAD dataset and 7 275 DEG of the GSE68468 dataset were screened out, and totally 237 differentially co-expressed genes were obtained. Ten core differentially co-expressed genes were obtained by the MCC calculation method of the PPI network, which were chloride channel accessory 1 ( CLCA1), mitogen-activated protein kinase 3, glucagon ( GCG), solute carrier family 26 member 3 ( SLC26 A3), nuclear receptor subfamily 1 group H member 4 ( NR1 H4), fatty acid binding protein 1 ( FABP1), guanylate cyclase activator 2A ( GUCA2 A), uridine diphosphate glucuronosyltransferase family 2 member A3 ( UGT2 A3), carnitine palmitoyltransferase 2 ( CPT2) and membrane spanning 4-domains A12 ( MS4 A12). Compared with those of the normal tissues, CLCA1, GCG, SLC26 A3, NR1 H4, FABP1, GUCA2 A, UGT2 A3, CPT2 and MS4 A12 of colon cancer tissues of the TCGA-COAD dataset were all down-regulated (all P<0.05). Among them, the overall survival time and disease-free survival time of patients with colon cancer with high expression of CLCA1 were both longer than those with low expression (both P<0.05). The results of immunohistochemical staining also verified the accuracy of the results at the protein level. Conclusions:CLCA1 may play a key role in the development of colon cancer, and it can be used as a potential biomarker for further diagnosis and treatment.
5.Expression of the transforming growth factor beta-induced gene in human corneal tissue and cell in vitro
Jing-yi, NIU ; Jing, LIU ; Lian, LIU ; Yi-yang, L(U) ; Jian-su, CHEN ; Jin-tang, XU ; Jing-xiang, ZHONG
Chinese Journal of Experimental Ophthalmology 2012;30(1):29-32
Background Clinical studies indicated that the pathogenesis of most corneal dystrophy is associated with the mutation of the transforming growth factor beta-induced (TGFBI) gene.However,the molecular mechanism of mutated TGFBI gene in corneal dystrophy is unclear. Objective The present study was to investigate the expression of the TGFBI gene in human corneal tissue and cells in vitro.MethodsHuman corneal epithelial cells and keratocytes were cultured and passaged,and donor corneal tissue was obtained for the section preparation.RT-PCR was used to detect the expression of TGFBI mRNA in human corneal tissue and cells.Immunofluorescence was used to test the expression of the TGFBI protein in the human corneal tissue,and immunohistochemistry was used to test the expression of the TGFBI protein in human corneal epithelial cells and corneal stromal cells.ResultsRT-PCR analysis showed that TGFBI mRNA could be detected as a 1274 bp band in human corneal tissue and corneal stromal cells,but no TGFBI mRNA was observed in corneal epithelial cells.Immunofluorescence assay revealed that corneal stromal cells were positive ly expressed for the TGFBI protein,but the corneal epithelial cells did not express the TGFBI protein.Immunohistochemistry indicated that the expression of TGFBI was detected the red fluoressence in the cytoplasm of corneal stromal cells;however,no positive response was found in corneal epithelial cells.ConclusionsThe expression of the TGFBI gene occurs in human corneal stromal cells but not in the corneal epithelial cells.This result might be of helpful for studying the function and role of TGFBI gene in pathogenesis of corneal dystrophy.
6.Effects of Saussurea involucrata extract pretreatment on the expression of the Toll-like receptor 4/nuclear factor-κB in focal cerebral ischemia/reperfusion in mice
Hui DANG ; Shanjiang AI ; Juan BU ; Jian LI ; Jing SHA ; Yan JING ; Yi ZHU
International Journal of Cerebrovascular Diseases 2012;20(9):690-695
Objective To investigate the effects of Saussurea involucrata extract pretreatment on the expression of the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) in focal cerebral ischemia/reperfusion in mice and its possible neuroprotective mechanism.Methods Seventy-two Kunming mice were randomly divided into four groups:sham operation,saline,Saussurea involucrata extract,and edaravone groups (n =18 in each group).Saussurea involucrata extract 0.8 g/kg was given intraperitoneally in the Saussurea involucrata extract group; edaravone 3 mg/kg was given in the edaravone group; and the same volume of saline was given in the saline group.A model of middle cerebral artery occlusion (MCAO) was induced after 7 days of continuous injection.Cerebral infarct volume was determined by 2,3,5-triphenyltetrazolium staining.Immunohistochemical staining was used to detect TLR4-positive cells in ischemic brain tissue.Reverse transcriptase polymerase chain reaction was used to detect the expression of TLR4/NF-κB mRNA.Results The cerebral infarct volume in mice in the saline,Saussurea involucrata extract and edaravone groups was 131.55± 28.25 mm3,84.10 ±13.92 mm3 and 65.10 ± 6.78 mm3,respectively.There were significant difference (F =10.158,P =0.012).The infarct volume in the Saussurea involucrata extract group (P =0.020) and edaravone group (P0.005) was significantly less than that in the saline group,and there was no significantly difference between the 2 groups.The numbers of cortex and TLR4 positive cells in hippocampus area at the ischemic sides in the saline group were significantly more than those in the sham operation group (all P <0.001).The numbers of positive cells of cortex and TLR4 in the Saussurea involucrata extract group and the edaravone group were significantly decreased compared to the saline group (all P < 0.05),and there was no significant differences between the Saussurea involucrata extract group and the edaravone group.The expressions of TLR4,p50,and p65 mRNA in the saline group were significantly up-regulated compared to the sham operation group (all P =0.000).Saussurea involucrata extract could significantly down-regulate the expressions of TLR4,p50,and p65 mRNA at 24 hours after ischemia/reperfusion (all P =0.000).Edaravone could significantly down-regulate the expressions of TLR4 and p65 mRNA (all P =0.000) and it had a down-regulated trend for the expression of p50 mRNA (P =0.053); while there was no significant difference in the expressions of TLR4 and p65 mRNA between the Saussurea involucrata extract group and the edaravone group.Conclusions Saussurea involucrata extract pretreatment may significantly reduce the cerebral infarct volume,down-regulate the expressions of TLR4 and NF-κB subunit,and play a neuroprotective effect by inhibiting inflammatory response after ischemia.
7.Clinical application of 64-slice spiral CT pulmonary angiography in diagnosis of pulmonary embolism
Jing GONG ; Jian-Ming TIAN ; Yi XIAO ; Min-Jie WANG ; Jian-Zhong GUAN ; Xiao-Ming LI ; Jing-Li TIAN ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective:To evaluate the clinical application of 64-slice spiral computer tomography pulmonary angiography (CTPA)in diagnosis of pulmonary embolism(PE).Methods:Sixty-two patients suspected of PE were examined by 64-slice spiral CTPA.The image findings combined with their clinical data were retrospectively analyzed.Results:Twenty-four of the 62 patients were confirmed to have PE by clinical data,laboratory examination and follow-up examination.64-slice spiral CTPA discovered 152 involved branches in the 24 PE patients,including 4 branches in left and right pulmonary trunk,52 in lobar pulmonary arteries,82 in segmental pulmonary arteries,and 14 in subsegmental arteries.Four types of PE were detected in our group,including eccentric filling defect in 58 branches,central filling defect in 49 branches,total occlusion of the pulmonary arteries in 21 branches,and mural embolism of host artery in 24 branches.The diagnosis accuracy of 64-slice spiral CTPA in the present group of patients was 100%,with no missed diagnosis and misdiagnosis.Besides,64-slice spiral CTPA could reflect the location,morphology,involvement and degrees of PE.Conclusion:64-slice spiral CTPA is a rapid,accurate and non-invasive diagnostic approach for PE.It is the first choice in clinical screening of PE and may serve as a gold standard for diagnosis of pulmonary embolism.
8.Application of quantum dots labeling technique in induced pluripotent stem cells
Mei-hua, TAN ; Jian-su, CHEN ; Jian, CHEN ; Jing, WU ; Zhi-yi, ZHAO ; Ying, DAI ; Shan-yi, LI
Chinese Journal of Experimental Ophthalmology 2013;31(9):819-822
Background The multipotent differentiation features of induced pluripotent stem cells (iPSCs) offer a new option for cell replacement therapy of many clinical diseases.In ophthalmology,iPSCs are a good model in studying the pathogenic mechanism of degenerative ocular diseases.A better identification method for iPSCs is critical for analyzing the in vivo biological characteristics of iPSCs.Objective This study was to investigate the feasibility and stability of labeling iPSCs with quantum dots.Methods Human umbilical mesenchymal stromal cells-iPSC lines were cultured and amplified on matrigel,and the characteristics of iPSCs were evaluated by immunofluorescence.Different concentrations (5.0,7.5 and 10.0 nmol/L) of quantum dots with a CdSe/ZnS nuclear shell structure were used to label iPSCs after passaging and proliferation.The labeling outcome was observed with a three-dimensional deconvolution real-time live cells imaging system.The labeled iPSCs were subsequently cultivated,and then changes in fluorescence intensity were examined 7 days after the first and the second passaging of iPSCs.Results iPSCs were observed to grow in a clonal manner under the inverted microscope.The iPSC markers,OCT4 and Nanog,were detected by immunofluorescence.With increasing concentrations of quantum dots,the fluorescence intensities representing the levels of OCT4 and Nanog in iPSCs were gradually elevated,with optimal levels of fluorescence observed at a concentration of 10 nmol/L of quantum dots.The fluorescent labeling of OCT4 and Nanog in iPSCs remained and weakened gradually till day 7 even after the second passage.Conclusions Quantum dots labeling could be used to track iPSCs in a dose-independent manner.The fluorescent signal from the quantum dots labeling the iPSCs lasts 2 weeks at least.
9.Construction of eukaryotic plasmid expressing human transforming growth factor beta-induced gene and its influence on human corneal epithelial cell
Jing-yi, NIU ; Jing, LIU ; Xiao-xia, LI ; Jian-su, CHEN ; Jin-tang, XU ; Jing-xiang, ZHONG
Chinese Journal of Experimental Ophthalmology 2011;29(12):1071-1076
Background The human transforming growth factor beta-induced gene (TGFBI) is the first determined pathogenic gene to corneal dystrophy.But the molecular genetic mechanism is completely unknown.The study of concerning role of TGFBI is very important for us understand the physiological function of cornea,and the pathogenesis of corneal dystrophy.Objective The vector of human transforming growth factor beta-induced gene (TGFBI) in eukaryotic expression was constructed and transfected into the human corneal epithelial cells in order to explore its influence on the growth of human corneal epithelial cells.Methods Total RNA was extracted from normal donor cornea tissue and cDNA was obtained by reverse transcription.TGFBI cDNA was synthesized by reverse transcription-PCR and cloned into pCMV-N-HA vector and identified by sequencing with PCR and EcoRV,XhoI double restriction endonuclease.The cells were grouped into recombinant pCMV-N-HA-TGFBI plasmid group,pCMVN-HA plasmid group,non-transfected group and pGFP-C2 transfected group.The recombinant pCMV-N-HA-TGFBI plasmid was transfected to human corneal epithelial cells and identified by observing the expression of enhanced green fluorescence protein(EGFP) in the cells.The TGFBI mRNA and proteins were harvested from the cells for real-time PCR analysis and Western blot assay respectively in 58 hours after transfection.The growth of the transfected cells was assessed by Cell Counting Kit-8.The expressions of matrix metalloproteinase(MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) proteins and their mRNA in transfected cells were detected using SYBR fluorescence realtime PCR analysis and Western blot assay.Results The sequencing result of pCMV-N-HA-TGFBI positive clone plasmid showed that amplified TGFBI eDNA inserted into the vector at the correct sequence.EGFP was expressed in transfected cells in 48 hours after transfer of pGFP-C2 with the transfer efficacy 70%.The expression intensity of TGFBI mRNA was significantly higher in recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group,and TGFBI protein was expressed in recombinant pCMV-N-HA-TGFBI plasmid group.No significant difference was found in the A450value among recombinant pCMV-N-HA-TGFBI plasmid group,pCMV-N-HA plasmid group and non-transfected group ( F=3.34,P>0.05 ).The mRNA level of MMP1,MMP3in the transfected cells was significant elevated but that of TIMP1 was declined in the recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group (all P < 0.05 ).Meanwhile,the expressions of MMP1,MMP3 and TIMP1 proteins appeared the same tendency( all P<0.05).Conclusions Eukaryotic expression vector harboring human TGFBI eDNA can be successfully constructed and efficiently overexpressed in human corneal epithelial cells.TGFBI gene is involved in the physical and pathological conditions of human corneal epithelial cells by regulating the activity of MMP1,MMP3 and TIMP1.The results offer a new approach for the study of the role of TGFBI in pathogenesis of corneal transparency.
10.Synthesis and characterization of urocanic acid-coupled chitosan as gene vector.
Yi-ping LI ; Jing YAO ; Jian-ping ZHOU ; Wei WANG
Acta Pharmaceutica Sinica 2008;43(12):1233-1238
A new nonviral gene vector--urocanic acid-coupled chitosan (UAC) was prepared by the reaction of the activated urocanic acid (UA) with the amine group on the chitosan (CTS). The structure of UAC was confirmed with FT-IR, 1H NMR and element analysis. The influencing factors of substitution values were studied by orthogonal test, and the substitution values of UAC increased with the prolongation of activating time of UA and the increasing ratio of UA to CTS. The condensation ability and the resistance to DNase I of UAC/pDNA were evaluated by agarose gel electrophoresis, and UAC showed good condensation ability with pDNA, well protecting pDNA from the degradation by DNase I. The particle size and zeta potential were evaluated by zetasizer, and the results showed that the UAC/pDNA complex was well stable and could easily enter into cells. The transfection studies were performed with HepG2 cells in vitro. It showed that the in vitro transfection of UAC/pDNA was efficient in HepG2 cells and could express more green fluorescent proteins than that of CTS. So the UAC is easy to prepare and a promising non-viral gene vector.
Chitosan
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administration & dosage
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chemical synthesis
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metabolism
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DNA
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genetics
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metabolism
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Deoxyribonuclease I
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metabolism
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Drug Delivery Systems
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Genetic Therapy
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methods
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Genetic Vectors
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Hep G2 Cells
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Humans
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Particle Size
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Plasmids
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Transfection
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Urocanic Acid
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administration & dosage
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chemical synthesis
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metabolism