1.Research Progress on Antiviral Activity of Interferon-induced Transmembrane Proteins.
Yongkun CHEN ; Wenfei ZHU ; Yuelong SHU
Chinese Journal of Virology 2016;32(2):222-228
Interferon-induced Transmembrane Proteins (IFITMs) were identified through small interference RNA (siRNA) screening method in 1980s. The antiviral properties of the IFITMs were firstly discovered in 1996. Recently, its antiviral effect and mechanism have become a research hotspot. Many studies have shown that IFITM can inhibit the replication of multiple pathogenic viruses, including influenza A virus (IAV), Human Immunodeficiency Virus (HIV-1), hepatitis C virus (HCV), Ebola virus (EBOV), West Nile virus and so on. IFITMs inhibit the replication of virus in the early stage of the viral life cycle, which occurred before the release of viral genomes into the cytosol. Recent studies indicate that IFITM proteins could block viral replication by mediate viral membrane fusion. However, the mechanism is still under investigation. Here we review the discovery and characterization of the IFITM proteins, elucidate their antiviral activities and the potential mechanisms.
Animals
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Humans
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Interferons
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genetics
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immunology
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Membrane Proteins
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genetics
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immunology
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Virus Diseases
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genetics
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immunology
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virology
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Viruses
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genetics
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immunology
2.The effects of gossypol acetic acid on DNA methylation level and the mRNA expression of hMLH1 gene in human tongue carcinoma Tca8 1 1 3 cells
Wenfei CHEN ; Shuai FU ; Wanyuan YUE ; Yong WU
Journal of Practical Stomatology 2014;(4):551-555
Objective:To study the effects of gossypol acetic acid(GAA)on the methylation level and the mRNA expression of hM-LH1 in human tongue cancer Tca8113 cells.Methods:Tca8113 cells were treated by GAA at various doses for 24 h,48 h and 72 h respectively.MTT assay was used to detect the cell proliferation.Nested methylation specific PCR(nMSP)was used to detect methyl-ation level of hMLH1 .Real-time fluorescence quantitative PCR(RFQ-PCR)was applied to investigate the mRNA expression of hM-LH1 gene.Results:GAA inhibited the proliferation of Tca8113 cells dose-and-time dependently,decreased the DNA methylation level of hMLH1(P<0.05)and increased hMLH1 mRNA expression in Tca8113 cells(P<0.05).Conclusion:GAA can suppress proliferation of Tca8113 cells by demethylation of hMLH1 gene and increase of hMLH1 mRNA expression.
3.The clinical significance and correlation of Notch1/DLL4 pathway and VEGF in invasion and metastasis of gastric carcinoma
Anhua ZHANG ; Huawen SUN ; Jinsong SU ; Zhonghui CUI ; Wenfei CHEN
Chinese Journal of General Surgery 1997;0(04):-
Objective To investigate the expression of Notch1 and its ligand DLL4 in human gastric carcinoma tissues and its correlation with tumor angiogenic metastasis.Methods Immunohistochemical SP method was used to detect the expression of Notch1,DLL4 and VEGF in 45 gastric carcinoma tissues and paired adjacent normal gastric mucosa,and the relationship between them and clinico-pathological parameters were analyzed.Results The positive expression rate of Notch1,DLL4 and VEGF in gastric carcinoma were higher than that in normal gastric mucosa(P
4.The effect of low glycemic index diet and exercise on plasma glucose and lipid metabolism in patients with newly diagnosed type 2 diabetes
Wenfei ZHONG ; Kaining CHEN ; Yi LI ; Tuanyu FANG ; Huachuan ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2011;18(4):445-447
ObjectiveTo evaluate the effect of low glycemic index(LGI)diet and exercise on plasma glucose and lipid profiles in newly diagnosed type 2 diabetic patients. MethodsSeventeen newly diagnosed type 2 diabetic patients with FPG ≤ 10mml/L treated by LGI diet and exercise only for two months.Fasting plasma glucose (FPG),2 hours postprandial glucose(2hPG),glycosylated hemoglobin A1 C(GHbA1C),and lipid profiles were measured.The results of FPG,2hPG,GHbA1C,and lipid profiles were compared. ResultsTwo months after treatment,the level of fasting glucose(6.19 ± 0.60)mmol/L,postprandial 2h plasma glucose(8.59 ± 0.90)mmol/L,TG(1.15 ± 0.45)mmol/L,TC(4.98 ± 0.77)mmol/L,LDL(3.20 ± 0.71)mmol/L were significantly lower than (7.84 ± 1.19)mmol/L,(13.97 ± 3.35)mmol/L,TG(1.79 ± 0.75)mmol/L,TC(5.46 ± 0.27)mmol/L,LDL (3.57 ± 0.28)mmol/L,HDL(1.59 ± 0.30)mmol/L was significantly higher than(1.42 ± 0.26)mmol/L,the differences were statistically significant(all P<0.05);HbA1c(6.49 ± 0.57)% was slightly lower than(7.29 ±0.77)%,but the difference was not significant(P>0.05);No hypoglycemia was observed during the treatment. ConclusionThe exellent glycemic control and improvement of lipid profile could be achieved by low glycemic index diet and exercise only.Furthermore,no hypoglycemia occurred during the treatment.
5.Effects of gossypol acetic acid on the proliferation and methylation level of the human MutL homologue 1 gene in human tongue carcinoma cell line Tca8113.
Shuai FU ; Yong WU ; Qingfang PENG ; Wenfei CHEN
West China Journal of Stomatology 2014;32(1):91-95
OBJECTIVEThis paper aims to study the effects of gossypol acetic acid (GAA) on proliferation and methylation level of human MutL homologue 1 (hMLH1) gene in human tongue cancer cell line Tca8113.
METHODSThe MTT assay was used to determine the effects of the acid on the proliferation inhibition in Tca8113 cells treated with different GAA concentrations. Nested methylation-specific polymerase chain reaction (nMSP) was used to detect the change in the methylation level of hMLH1 after 48 and 72 h with 30 and 15 micro mol L(-1) GAA treatment.
RESULTSMTT assay results showed the growth and proliferation inhibition of Tca8113 cells in the experimental GAA group after 24 h to 72 h of GAA treatment. The nMSP results indicated that the average optical density of hMLH1 in the Tca8113 cells significantly changed after the GAA treatment (30 micro mol L(-1) GAA for 48 h and 15 micro mol L(-1) for 72 h) (P<0.05) compared with that of the control group.
CONCLUSIONGAA does not only inhibit Tca8113 proliferation but also has a demethylation effect on the hMLH1 gene. These phenomena may be part of an underlying tumor-suppression mechanism of GAA.
Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Gossypol ; analogs & derivatives ; Humans ; Methylation ; Tongue Neoplasms
6.Effects of Gossypol Acetic Acid on Protein and mRNA Expressions of hMLH1 Gene in Human Tongue Carcinoma Cell Line Tca8113
Wenfei CHEN ; Shuai FU ; Wanyuan YUE ; Yong WU
Journal of Kunming Medical University 2014;(1):8-11
Objective The purpose of this study was to investigate the effects of gossypol acetic acid (GAA) on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods (1) Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. (2) Real-time fluorescence quantitative PCR (RFQ-PCR) was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results (1) Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h ( <0.05) . (2) The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h ( <0.05) . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.
7.Allele-31 C>T regulates binding activity to IL-1βgene promoter of nuclear transcription factor C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection
Guoliang ZHANG ; Rongrong ZOU ; Lingling WANG ; Wenfei WANG ; Mingxia ZHANG ; Guilin YANG ; Xinchun CHEN
Chinese Journal of Clinical Infectious Diseases 2016;9(2):180-185
Objective To investigate the effects of allele-31 C>T on the binding activity to IL-1βpromoter of the nuclear transcription factor C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection.Methods The electrophoretic mobility shift assay ( EMSA) was performed to explore whether the nuclear transcription factor C/EBPβand PU.1 could bind to -31 region in IL-1βpromoter.The C/EBPβ-and PU.1-expressing vectors were constructed and co-transfected into HeLa cells with IL-1βpromoter luciferase vector.The expression of C/EBPβand PU.1 was confirmed using Western blotting assay, and the promoter activity was determined using Dual-Glo Luciferase system under various transfection conditions. Lentivirus-mediated RNA interference was used to explore the effects of C/EBPβand PU.1 on IL-1βexpression.GraphPad Prism 5.0 was used for data analysis.Results EMSA results showed that both C/EBPβand PU.1 could bind to -31 region in IL-1βpromoter.Both C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection could increase IL-1βpromoter activity, especially for the -31 T allele (t=22.33 and 7.98,P<0.01), and there was a synergy on the promoter activity between C/EBPβand PU.1.The promoter activity decreased significantly when C/EBPβand/or PU.1 were silenced by lentivirus-mediated RNA interference (q=5.79, 6.23 and 11.66,P<0.01).Conclusion The allele-31 C>T can induce IL-1βpromoter activity and gene transcription through regulation of binding activity to C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection.
8.Serial research of traceability and harmonization of thyroid-stimulating hormone detection approved by IFCC
Xiang YU ; Wenfei CHEN ; Xiaoliu YU ; Ming JIANG ; Ke LI ; Zhiguang YU
International Journal of Laboratory Medicine 2016;37(6):751-753
Objective To implement the harmonization of thyroid-stimulating hormone(TSH) of Mindray assay system by par-ticipating in harmonization research program of International Federation of Clinical Chemistry (IFCC)-Standardization of Thyroid Function Tests(C-STFT ) .Methods A total of three combinations of TSH reagents and calibrators were used to measure 20 serum samples of healthy human .Within-run precision and batch-to-batch variation were assessed .The concept of harmonization was dem-onstrated by comparing our test results with All Procedure Trimmed Mean (APTM ) provided by IFCC and recalibrating with Mater Calibrators .Results The within-run precision was 1 .96% ,batch-to-batch variation was 0 .47% - 1 .15% ,depending on the level of TSH analyte .There existed a positive bias compared to APTM values .After recalibration with Mater Calibrators and Passing &Bablok regression ,the slope of method comparison was 1 .0 ,and correlation coefficient was more than 0 .975 .Conclusion By using a panel of real human specimen and recalibration based on APTM ,the test results of Mindray assay system could be harmonized with mainstream manufacturers globally .
9.Construction a shuttle vector containing AaCPR100A epidermal protein gene of Aedes aegypti with two spore-producing-dependent promoters in Bacillus thuringiensis by in-fusion cloning
MA Chenxin ; ZHANG Yingxin ; LIU Sihan ; HE Jiali ; CHEN Jiukai ; ZHANG Wenfei ; LIAO Chenghong ; HAN Qian
China Tropical Medicine 2023;23(11):1141-
Abstract: Objective To construct a shuttle vector pHT315-AaCPR100A with two spore-producing-dependent promoters and the target gene AaCPR100A in Escherichia coli-Bacillus thuringiensis. Methods The forward promoter of Cry3A, named Pro-1 (+), was amplified by PCR using pSVP27A plasmid as the template, and the target gene AaCPR100A was amplified using Aedes aegypti RNA reverse conversion cDNA as the template. The plasmid pHT315 was linearized by digestion with Hind Ⅲ and Sal Ⅰ. The forward promoter and the target gene were inserted into the linearized vector pHT315 successively by in-fusion cloning according to the transcription direction. The synthesized plasmid containing the Cry3A reverse promoter sequence was used as the template, and the Pro-1 (-) reverse promoter was amplified by PCR. The intermediate vector containing the forward promoter and the target gene was linearized by EcoR I restriction enzyme, and the reverse promoter was inserted downstream of the target gene by in-fusion cloning in the direction of transcription. Results By agarose gel electrophoresis, the forward promoter, target gene AaCPR100A and reverse promoter bands were clear and of good quality, which could be used for in-fusion cloning experiments. The two spore-producing-dependent promoters and target gene fragments were connected by In-fusion cloning. The recombinant vector pHT315-AaCPR100A was verified by PCR. The forward promoter, target gene fragment and reverse promoter were successfully amplified in the recombinant vector. Nucleotide sequencing verified that the sequencing results of the bidirectional promoter sequence and the target gene sequence were basically consistent with the sequence alignment results, which met the requirements of the construction of vector elements and proved that the recombinant vector was successfully constructed. Conclusions Based on the above results, this study proves that the recombinant shuttle vector with two spore-producing-dependent promoters can be successfully constructed by in-fusion cloning technology, laying the foundation for the construction of engineered Bacillus thuringiensis expressing dsRNA of AaCPR100A.
10.Prognostic Analysis of Skull-base Invasion of Nasopharyngeal Carcinoma Based on Magnetic Resonance Imaging
Lei CHEN ; Wenfei LI ; Lizhi LIU ; Yanping MAO ; Linglong TANG ; Ying SUN ; Aihua LIN ; Li LI ; Jun MA
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(2):258-264
[Objective]To evaluate the prognostic value of skull-base invasion of nasopharyngeal carcinoma(NPC)based on magnetic resonance imaging(MRI).[Methods]A total of 924 patients who were diagnosed with NPC between 2003 and 2004,had undergone MRI scan and received mdiothempy as their primary treatment,and had no distant metastasis were included in this study.MRI images and medical records were analyzed retrospectively.All the 924 eases.patients who developed skull-base invasions based on MRI,315 patients with T3 disease and 227 patients with T2 disease were selected for analysis.The staging was according to the sixth edition of the American Joint Commission on Cancer(AJCC)staging system.[Results]Incidence of skullbase invasion according to MRI was 55.4%.Of 924 cases.skull-base invasion on MRI was not an independent prognostic factor for overall survival(OS)and distant metastasis-free survival(DMFS),but was a marginally significant independent prognostic factor for local relapse-free survival(LRFS),P=0.068.Grading of MRI-detected skull-base erosion according to the site of invasion was an independent prognostic factor for OS(P=0.002 and P=0.005)and DMFS(P=0.001 for both)in the 512 patients with skull-base invasions and 315 patients with T3 disease.Severe-grade of skull-base invasion on MRI was an independent prognostic factor for OS and DMFS in the 924 patients(P < 0.001 for both).No significant differences were observed on OS,LRFS,and DMFS between T2a patients and T3 patients with low-grade of MRI-deteeted skull-base involvement.[Conclusions]Skull-base invasion based on MRI is not an independent prognostic factor for NPC.However,severe-grade of invasion according to the site of involvement has positive prognostic value.