1.The effects of let-7c-1 on the learning and memory of epileptic rats induced by PTZ
Yayun LIAO ; Xixia LIU ; Yiqing HUANG ; Yuhan LIAO ; Yuan WU
Chinese Journal of Nervous and Mental Diseases 2017;43(4):239-244
Objective To explore the effect of let-7c-1 on the learning and memory of PTZ-induced epileptic rats and its relevant mechanism.Methods A model of temporal lobe epilepsy (TLE) was induced via PTZ kindling in SD male rats.The epileptic rats were divided into epilepsy group,agomir-control group,let-7c-1 agomir group (12 rats for each).Twelve rats were served as a negative control group.The behavior and the expression levesl of let-7c-1,Bcl-2 protein and Caspase3 were evaluated at 28 days following PTZ.Results Compared to the negative group,the escape latency of epilepsy group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P<0.05).However,those parameters were not significantly different between the epilepsy group and the agmoir-control group (P>0.05).Compared to the agmoir-control group,the escape latency of let-7c-1 agomir group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P< 0.05).The expression levels of let-7c-1 and let-7c-1 were 1.35±0.32 in agmoir-control group and 62.53±21.01 in agomir group (F=50.97,P<0.05).The expression levels of let-7c-1 were higher in let-7c-1 agomir group than in other groups (P<0.05).Compared to the negative group,the expressions of Bcl-2 protein in other groups were decreased (P<0.05) and the Caspase3 protein were increased (P<0.05).Compared to the agomir-control group,the expression of Bcl-2 protein was significantly decreased and the expression of Caspase3 protein was significantly increased in let-7c-1 agomir group (P<0.05).Conclusions The present study shows that let-7c-1 may impair the learning and memory of PTZ-induced epileptic rats through decreasing the Bcl-2 protein and increasing Caspase3 protein in the hippocampus.
2.Detection of Salmonella in Food with Phage
Gaoli WU ; Tao YANG ; Lipeng LIU ; Qiong ZHANG ; Xixia LIU ; Xiaolan TANG
Journal of Kunming Medical University 2013;(11):12-14
Objective To investigate the pollution condition of Salmonella in food with phage. Methods Salmonella in 413 samples of food were detected by the diagnosis of Salmonella phage with biochemical and serological identification. Results 119 Salmonella were detected in 99 positive samples and the isolating rate was 24%. Conclusion The disservice of Salmonella is mainly from the meat food and eggs. The detection method of phage is fast,convenient and reliable.
3.Cell-specific roles of domains I and II of HCV 5'untranslated region in the translation initiation activity.
Xiaoye HUANG ; Lisha LIU ; Guangjing CUI ; Xixia LIU ; Meitong LIU ; Qiongshan MA ; Shuiping LIU
Journal of Southern Medical University 2014;34(12):1826-1829
OBJECTIVETo investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines.
METHODSThe eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed.
RESULTSDeletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46% in L-02 cells and increased the translational activity by 46% in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51% in HeLa cells, but increased the translational activity by 40% in L-02 cells, 60% in C6 cells and 135% in 293T cells.
CONCLUSIONSDomain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.
5' Untranslated Regions ; Genes, Reporter ; HeLa Cells ; Hepacivirus ; genetics ; Humans ; Luciferases ; Plasmids ; Protein Biosynthesis ; genetics ; RNA, Viral ; genetics ; Transfection
4. Evaluation of 5′-untranslated region amplification and sequencing for enterovirus serotypes identification diagnosis
Shihuan TANG ; Zhenghua XIE ; Duoduo LIU ; Ying YUAN ; Manjun CHEN ; Xiaodi FAN ; Xixia DING ; Nan YU
Chinese Journal of Experimental and Clinical Virology 2018;32(5):488-491
Objective:
To evaluate an assay permitting amplification of target 5′-untranslated region (5′-UTR) sequences directly from clinical specimens and distinction among serotypes of enterovirus (EV).
Methods:
A total of 518 rectal swabs and 148 nasal swabs tested positive by pan-enterovirus real-time PCR were collected. 5′-UTR and the viral protein 1 (VP1) gene fragments were amplified and sequenced separately for serotyping. The inconsistent samples by 5′-UTR and VP1 serotyping were further determined by using the serotype-specific RT-PCR.
Results:
A total of 553 (83.0%) samples were detected by 5′-UTR serotyping and 318 (47.7%) were detected by VP1 serotyping in all 666 positive specimens, and there was significant difference in the detection rates between two methods in rectal and nasal swabs (
5.Construction and expression of anti-clenbuterol single chain Fv recombinant vector.
Hong WANG ; Yan LIANG ; Jingyi YANG ; Xixia LIU ; Hongbin ZHANG ; Hongtao LEI ; Yudong SHEN ; Yuanming SUN
Chinese Journal of Biotechnology 2008;24(8):1470-1474
To construct the recombinant vector pBV220-scFv and express anti-clenbuterol (CBL) scFv antibody in Escherichia coli, we amplified the scFv gene using plasmid pCANTABSE-CBL as a template, recombined it with pPICZalphaA, then amplified the scFv-His-tag gene from plasmid pPICZalphaA-scFv and linked it with expression plasmid pBV220. We identified the recombinant plasmid by restrictive enzyme digestion, PCR amplification and sequence analysis. Finally, we transformed the recombinant vector into E. coli DH5alpha that was temperature-induced and expressed recombinant protein. We identified the recombinant protein by SDS-PAGE, Western blotting and indirect competitive ELISA. The results show that recombinant plasmid pBV220-scFv contained the inserted fragment with highest homology about 99.8%. The expression of scFv induced by temperature show 37 kD Mw and anti-His-tag mAb recognized-activity by SDS-PAGE and Western blotting respectively, and could competitively combine with CBL, the IC50 is 4.55 ng/mL. The recombinant plasmid pBV220-scFv is constructed and expresses the scFv gene of CBL in E. coli successfully. This study suggests the corresponding immunoassay methods could be set up by the recombinant scFv.
Antibodies
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immunology
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Clenbuterol
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immunology
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Cloning, Molecular
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Genetic Vectors
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genetics
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Immunoglobulin Fragments
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biosynthesis
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genetics
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immunology
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Immunoglobulin Variable Region
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biosynthesis
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
6. Clinical features of 406 cases of dengue fever in Guangzhou from 2015 to 2018
Zhenghua XIE ; Duoduo LIU ; Shihuan TANG ; Ying YUAN ; Yue CHEN ; Manjun CHEN ; Xixia DING ; Nan YU
Chinese Journal of Experimental and Clinical Virology 2019;33(3):257-260
Objective:
By analyzing the clinical features of patients with dengue fever in Guangzhou from 2015 to 2018 to furnish the reference evidences for the diagnoses and treatment of dengue fever.
Methods:
A total of 406 dengue fever patients admitted to Zhujiang hospital during 2015 to 2018 were analyzed for the clinical manifestations and laboratory examination results, retrospectively. ZIKV, CHIKV and the serotypes of DENV were detected in some samples.
Results:
DENV serotypes were tested in 96 dengue fever patients and 69 cases were positive. Among them, 58 cases (84.1%) were DENV-1, 10 cases (14.5%) were DENV-2, 1 case (1.5%) was DENV-3, DENV-4 was negative and no co-infection with different serotypes of dengue virus was found. Of all the 406 patients, 371 (91.4%) were diagnosed as dengue fever and 35 (8.6%) were severe cases. The most common manifestations included fever, weakness and rash. Significantly higher incidence (