2.Construction of human embryonic kidney cells exhibiting human preproenkephalin gene expression
Feng BAI ; Baozhong YANG ; Zhaoxia XUE ; Yufei PAN ; Ping HUANG
Chinese Journal of Anesthesiology 2012;32(6):673-675
ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.
3.Correlation between overexpression of TLR4/MyD88 signaling and intestinal graft-versus-host disease after allogeneic bone marrow transplantation in mice
Shuai XING ; Xue ZHANG ; Xia HUANG ; Ping ZHOU
Chinese Journal of Organ Transplantation 2014;35(7):431-435
Objective To explore the expression of TLR4/MyD88/NF-κB signaling and its correlation with the progression of acute intestinal graft-versus-host disease (iGVHD) after allogeneic bone marrow transplantation in mice.Method Recipient BALB/c female mice were lethally irradiated and were reconstituted within 4-6 h with a transplant of bone marrow cells (1 × 107) and different amounts of splenocytes (1 × 107,n =12 or 2 × 107,n =12) from MHC-mismatched C57BL/6 donors to induce iGVHD,and 6 healthy BALB/c mice served as controls.A globe survey observation of GVHD by survival,clinical manifestation,and histological detection was performed.RT-PCR,immunohistochemistry,and Western blotting technology were used to detect the mRNA and protein levels of TLR4,MyD88 and NF-κB p65 in the small intestine tissue.Result The exacerbation of iGVHD was associated with the increasing dose of allogeneic spleen lymphocytes.The mRNA expression of TLR4,MyD88 and NF-κBp65 was increased as the iGVHD progressed.All of them in severe iGVHD model were significantly increased as compared with the healthy controls (P<0.05).The expression of corresponding proteins had the same tendency as mRNA.All of the three genes expression was not only positively correlated with each other,but also with the clinical GVHD score:TLR4 (R =0.814,P<0.001),MyD88 (R=0.828,P<0.001),and NF-κB p65 (R=0.568,P =0.034).Conclusion Excessive activation of TLR4/MyD88/NF-κB signaling pathway does exist in iGVHD,and the enhanced levels of gene transcription and translation are positively correlated with the deterioration of iGVHD.
4.Study on Fused Expression of the IgG-binding Domain of Streptococcal Protein G and Its IgG-binding Activity
Xue-Nian HUANG ; Yang XU ; Yan-Ping LI ;
China Biotechnology 2006;0(10):-
The IgG binding domain of Streptococcal Protein G which can selectively immobilizes the Fc regions of immunoglobulin G(IgG) is a kind of good material for oriented immobilization of antibodies in antibody microarrays.Here,genetically engineered three glutathione S-transferase(GST) fused proteins,bearing one,two and three B-Domains respectively(GST-GBx).The IgG-bindding ability of GST-GBx was investigated by ELISA.The data revealed that when the B-domain's quantity of GST-GBx is identical,the GST-GB3 is the most efficient protein among three GST-GBx protein both the capacity and sensibility of binding IgG.The GST-GB2 is the next one and GST-GB1 is the least one.Thus,the GST-GB3 has significantly predominance in comparison to GST-GB2 and GST-GB1.
5.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
6.Clinical study on 100 cases of severe acute pancreatitis in aged patients
Wei HUANG ; Ping XUE ; Tingting LIU ; Lei HUANG ; Dakai XIANG ; Lei WANG ; Qing XIA
Journal of Integrative Medicine 2007;5(3):268-71
OBJECTIVE: To analyze the clinical features of severe acute pancreatitis (SAP) in aged patients, and to explore the measures of its diagnosis and treatment. METHODS: The clinical data of 100 aged patients (55-85 years old) with SAP admitted from January 2003 to December 2005 were reviewed and compared with those of 221 non-aged SAP patients (11-54 years old) admitted at the same period. RESULTS: Totally 112 times of onset occurred in 100 aged patients, and the main causes for senile SAP were biliary diseases and hyperlipemia. Both acute physiology and chronic health evaluation II (APACHE II) and Ranson scores of the aged patients with SAP were higher than those of the control group (P<0.05); while there was no significant difference in Balthazar score between the two groups(P>0.05). The incidences of low serum calcium, low albumin and high aspartate aminotransferase (P<0.05), liver, kidney or brain impairment as well as the mortality were higher in the senile SAP group than those in the control group (P<0.05). CONCLUSIONS: Biliary disease and hyperlipemia are the main causes of senile SAP, which lacks characteristic clinical symptoms and is often associated with multiple complications such as infection and lung, gastrointestinal tract, liver, kidney, and brain impairment. Individualized differential treatment with integrated traditional Chinese and Western medicine based on stage classification is an effective approach.
7.Research on a tool for organizational culture assessment of public general hospitals in China
Ping ZHOU ; Jile CHANG ; Jinxing HUANG ; Lan LI ; Zhiliu TANG ; Jiayan HUANG ; Rong WU ; Di XUE
Chinese Journal of Hospital Administration 2011;27(6):433-436
Objective A scientific evaluation of hospital culture with the Dimension organizational culture model, in view of features of China's general public hospitals.Methods Based on Denison model, according to the characteristics of the public general hospitals in China, the authors developed a tool for organizational culture assessment (TOCA) by using the survey data from 87 hospitals in three provinces from the East, Central, and West areas in China.Results This tool, an evaluation scale, comprises the four cultural characteristics of direction, consistency, participation, and adaptability, as well as 13 cultural dimensions of social responsibility and competitive consciousness. The tool is tested as having good internal reliability and validity.Conclusion The TOCA provides hospital administrators with a tool for hospital culture evaluation, diagnosis and improvement.
8.Bioavailability and bioequivalence of erythromycin ethylsuccinate granules in healthy volunteers
Gui-Xiong ZENG ; Guo-Ping ZHONG ; Xue-Ding WANG ; Li-Hui HUANG ; Min HUANG ;
Chinese Journal of Infection and Chemotherapy 2006;0(04):-
Objective To study the bioavailability and bioequivalence of erythromycin ethylsuccinate granules in healthy male volunteers.Methods In a randomized two-period crossover study,20 healthy male volunteers received single 500 mg test and reference formulations of erythromyein ethylsuccinate granules.The plasma concentrations of erythromycin were assayed by microbiological method.Results The parameters of test and reference preparations were as follows:T_(max)(0.86?0.22)and (0.80?0.13)h,C_(max)(2.13?0.64)and(2.16?0.61)mg/L,t_(1/2)(2.04?0.2)and(1.97?0.4)h,AUC_(0-t)(4.96?1.73)and(4.63?1.52)mg?h/L,respectively.There was no significant difference between the two preparations.The rela- tive bioavailahility of the test granules was(109.1?22.8)%.Conclusions The two preparations of erythromycin ethylsucci- nate granules are bioequivalent.
9.Association of the progress of chronic periodontitis with interleukin-1B-511 genetic polymorphisms.
Xue-li WU ; Ping HUANG ; Xue-dong ZHOU
West China Journal of Stomatology 2006;24(6):523-526
OBJECTIVETo study the natural progress of different degree chronic periodontitis and its association with IL-1B-511 genetic polymorphisms. METHODS; 100 subjects with chronic periodontitis were selected and examined at baselined and in the 6 month and in 1 year on attatchment loss at 6 sites of each tooth. DNA samples were obtained with buccal swabbing technique and were further analyzed for IL-1B-511 genotype polymorphisms using PCR-RFLP-based method in all subjects.
RESULTSThe mean AL increases were 1.43 mm within 1 year. Among 100 subjects, 16 patients with moderate progression (0 mm < AL increase/a year < or = 1.0 mm), 84 patients with rapid progression of periodontal disease (AL increase/ a year > 1.0 mm). There was no significant difference for the distribution and frequency of IL-1B-511 genotype and alleles between the AL increase/ a year > 1.0 mm group and AL increase/a year < or = 1.0 mm group. The progression of periodontal disease (AL increase/a year > 1.0 mm group) was significantly higher in the non-severe chronic periodontitis group than in the severe group (P < 0.05). The percentage of molar was higher as far as the rapid-progress sites (AL increase > 2.0 mm both in the 6th and the 12th months examination) were concerned than that of premolar and anterior (P < 0.05). The number of progressed sites in the severe group was higher than the non-severe chronic periodontitis group (P < 0.05).
CONCLUSIONThe progress of chronic periodontits varies individually. No specific relationship was found between the progression of chronic periodontitis and IL-1 gene polymorphisms.
Adult ; Alleles ; Chronic Periodontitis ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukin-1 ; Male ; Middle Aged ; Periodontitis ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length
10.Expression of chemokine receptor CXCR4 in hypopharyngeal carcinoma and its significance
Guo-Hua SUN ; Yong-Xue ZHU ; Cai-Ping HUANG ; Yu WANG ; Ling ZHANG ; Xue-Chen WANG ; Qing-Hai JI ;
China Oncology 2000;0(06):-
Background and purpose:It was reported that chemokine receptor CXCR4 and its ligand stromal cell-derived factor 1(SDF-1)were involved in the proliferation,differentiation,and metastasis of tumor.This study was designed to observe the expression of chemokine receptor CXCR4 in hypopharyngeal squamous cell carcinoma tissue and study the relationship between the expression of chemokine receptor CXCR4 and different clinicopathlogical characteristics,and further to explore the clinical significance.Methods:For the detection of the expression of chemokine receptor CXCR4,43 primary hypopharyngeal squamous cell carcinoma tissues,27 normal hypopharyngeal tissues,34 lymph node metastastatic lesions and 9 normal lymph node lesions were detected by immunohistochemical method using rabbit anti-human CXCR4 polyclonal antibody.Results:The positive expression rates of CXCR4 in 43 hypopharyngeal carcinoma tissues and normal tissues were 95.3% and 22.2%,respectively(P