1.Transcriptome analysis of human neuroblastoma cells infected with Zika virus
Qiufeng YAO ; Lijian ZHANG ; Yaqi GAO ; Hao REN ; Zhaoling QIN ; Zhongtian QI
Academic Journal of Naval Medical University 2024;45(12):1508-1520
Objective To analyze the transcriptome data of Zika virus(ZIKV)-infected human neuroblastoma cells SH-SY5Y by bioinformatics method,and to identify the potential genes involved in the pathogenic mechanism of ZIKV.Methods SH-SY5Y cells were infected with ZIKV.The total RNA was extracted and the differentially expressed genes(DEGs)were screened by transcriptome sequencing.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed to predict the biological processes,molecular functions,and signaling pathways mainly involved in the DEGs,and the results were verified by quantitative polymerase chain reaction(qPCR).Results A total of 259 DEGs were identified,including 172 up-regulated genes and 87 down-regulated genes.GO functional enrichment analysis showed that the DEGs were mainly related to extracellular matrix,response to stimulus,antimicrobial humoral response,and developmental process.KEGG pathway enrichment analysis revealed that the DEGs were predominantly associated to inflammatory reaction and immune response.The qPCR validation results of DEGs were basically consistent with the transcriptome sequencing results.Conclusion The expression of genes involved in extracellular matrix,response to stimulus,and regulation of inflammatory reaction is significantly altered in SH-SY5Y cells after ZIKV infection,suggesting that ZIKV may cause neurological lesions by remodeling the extracellular matrix and regulating inflammatory reaction.
2.Proteomics and Phosphoproteomics Analysis of Effect of Retinoic Acid-Induced Protein 16 Knockout on Human Colon Cancer Cells
Yibo CHEN ; Gen MIAO ; Wen WANG ; Cuiling DING ; Zhongtian QI
Cancer Research on Prevention and Treatment 2024;51(10):820-830
Objective To analyze the differences in the expressions of the total and phosphorylated proteins in human colon cancer HCT116 cells after the knockout(KO)of retinoic acid-induced protein 16(RAI16)and explore the possible mechanism and related signaling pathways affecting its protein function in HCT116 cells.Methods HCT116 KO and WT cell proteins were collected and extracted,and the protein extraction efficiency was detected via a sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)experiment.After protein digestion,the peptides were labeled with TMT and analyzed via mass spectrometry.We used bioinformatics methods to analyze the identified differential proteins and differentially phosphory-lated proteins by using GO,KEGG,and STRING databases.Results The results of SDS-PAGE showed no evident protein degradation.In addition,some key bands were significantly different between the experi-mental and control groups.A total of 147 up-regulated and 230 down-regulated differential proteins were screened in accordance with the conditions of Foldchange≥1.5 or Foldchange≤1/1.5 and P<0.05.Meanwhile,106 up-regulated and 217 down-regulated phosphorylation sites were screened.GO enrichment analysis revealed that the differential proteins were mainly enriched in the composition of nucleoplasm,nucleus and cytoplasm,RNA binding,cadherin and chromatin,DNA repair,RNA splicing,and positive regulation of DNA as template transcription.The results of KEGG enrichment indicated that the differential proteins were mainly enriched in nucleocytoplasmic transport,spliceosomes,cell cycle,cell-cell tight junctions,viral carcinogenesis,microRNAs in cancer,etc.The protein interaction network mainly focused on DDX17,NCL,EEF2,CDK1,SSRP1,and SMARCC1.The statistical findings unveiled the up-regulated changes in the two omics of SKP1,ORC1,and BAD and the down-regulated changes in RBL1,RB1,CDK1,CDC6,MCM4,TFDP1,CHD4,and SNW1.Moreover,the phosphorylation differences were more significant than the protein differences.Conclusion RAI16 plays the possible crucial role in multiple biological functions and signaling pathways through key proteins,such as SKP1,ORC1,RB1,and CDK1,which affect the cell cycle and thereby the occurrence and development of cancer.
3.Research progress in site mutations of SARS-CoV-2 Omicron variants
Gen MIAO ; Zhiwei HE ; Yibo CHEN ; Cuiling DING ; Hao REN ; Ping ZHAO ; Zhongtian QI
Chinese Journal of Microbiology and Immunology 2023;43(3):173-181
SARS-CoV-2 Omicron variant (B.1.1.529) was first discovered in South Africa in November 2021 and has since become a mainstream strain worldwide. Omicron variant was defined as the fifth "variant of concern (VOC)" by World Health Organization on November 26, 2021. This paper illustrates the mutation trends of Omicron variants in terms of SARS-CoV-2 genome and protein structure as well as nucleic acid site mutations and amino acid site mutations, describes the features of Omicron mutation sites in terms of lineage comparison among the VOCs and Omicron sublineages, and further highlights the influences of Omicron site mutations from the aspects of immune escape, virulence and transmission ability. Moreover, this paper also reviews the development of direct antiviral agents, antibodies and vaccines, aiming to provide reference for further investigation.
4. Current status of treatment for 2019 novel coronavirus pneumonia
Naiwei ZHU ; Ping ZHAO ; Zhongtian QI
Chinese Journal of Microbiology and Immunology 2020;40(1):E002-E002
2019 novel coronavirus (2019-nCoV) is a new member of coronavirus family that can cause serious respiratory diseases after the emergence of severe acute respiratory syndrome-coronavirus (SARS-CoV) and middle east respiratory syndrome-coronavirus (MERS-CoV). At present, there is no specific antiviral drug targeting 2019-nCoV. In facing of the increasingly serious epidemic of 2019 novel coronavirus pneumonia and the urgent needs in drug treatment strategies, this paper reviewed the current research situation and progress in antiviral treatment for the newly identified disease.
5. Current status of treatment for 2019 novel coronavirus pneumonia
Naiwei ZHU ; Ping ZHAO ; Zhongtian QI
Chinese Journal of Microbiology and Immunology 2020;40(1):7-10
2019 novel coronavirus (2019-nCoV) is a new member of coronavirus family that can cause serious respiratory diseases after the emergence of severe acute respiratory syndrome-coronavirus (SARS-CoV) and middle east respiratory syndrome-coronavirus (MERS-CoV). At present, there is no specific antiviral drug targeting 2019-nCoV. In facing of the increasingly serious epidemic of 2019-nCoV pneumonia and the urgent needs in drug treatment strategies, this paper reviewed the current research situation and progress in antiviral treatment for the newly identified disease.
6. Advance in Zika virus infection-related myelitis
Gen MIAO ; Cuiling DING ; Yangang LIU ; Zhongtian QI
Chinese Journal of Microbiology and Immunology 2019;39(10):800-804
Zika virus belongs to the family Flaviviridae, genus
7.Identification of hepatitis B virus X-interactive proteins in human hepatocellular carcinoma cell lines HepG2 by using proteomics technique
Weihua LI ; Xiaohui MIAO ; Zhongtian QI ; Wu NI ; Kekai ZHAO ; Shiying ZHU
Chinese Journal of Infectious Diseases 2010;28(2):65-71
Objective To identify hepatitis B virus X-interactive proteins by comparative proteomics method and to understand the molecular mechanism of HBx in the development of hepatocellular carcinoma(HCC).Methods Two-dimensional gel electrophoresis(2-DE)was used to separate the total proteins of HBx-transfected human hepatoma cell lines HepG2-Px and its parental cell lines HepG2-P_0.PDQuest software was applied to analyze 2 DE images.The differentially expressed protein spots between the two cell lines were identified by matrix-assisted laser desorptionionization time of flight mass spectrometry(MALDI-TOF-MS)and electrospray ionization tandem mass spectrometry(ESI-Q-TOF).Then,the differential expression levels of some identified proteins were determined by Western blot.The data were compared using t test.Results The well-resolved,reproducible 2-DE patterns of HepG2-Px and HepG2-P_0 total proteins were established.A total of 32 differential proteins were identified in HepG2-Px cell,including 25 up-regulated proteins,such as heat shock protein(HSP)90AB1,Bcl-2 associated athanogene(BAG)-2,nucleophosmin(B23),chloride intracellular channel(CLIC)-1,matrix metalloproteinase(MMP)-3,melanoma antigen(MAGE)-12,and 7 down-regulated proteins,such as Wnt-5a.The differential expression levels of some proteins between the two cell lines were confirmed by Western blot analysis.Conclusions Most of the identified proteins are involved in many processes,such as transcription,signal transduction,cell proliferation,cell cycle regulator,apoptosis,DNA repair,metabolisms and immunity.These differential proteins may play a role in tumor genesis and HC development.The data are valuable for further study on the role of HBx in human hepatocellular carcinoma.
8.Experimental study of tissue-specific suicide gene therapy for lung adenocarcinoma.
Lijun ZHAO ; Qiang LI ; Chong BAI ; Yuguang LI ; Guangwen CAO ; Zhongtian QI
Chinese Journal of Lung Cancer 2004;7(4):290-293
BACKGROUNDTo study the effect and safety of tissue specific gene therapy of suicide gene for lung adenocarcinoma.
METHODSRetroviral vector of G1CEACDNa contained a carcinoembryonic antigen (CEA) promoter regulated cytosine deaminase expression cassete (CEA/CD). By means of infection of virus, tissue specific expressing vectors and non-specific expressing vectors were transfected into A549 cell, which was CEA-producing lung adenocarcinoma cell line and then xenografted in nude mice, and the anti-tumor effect was evaluated. Then the retrovirus was injected directly into the tumor mass on nude mice, and the sensitivity of the xenograft to G1CEACDNa/5-fluorocytosine (5-FC) and the side effects were observed.
RESULTS(1) After transfected and untransfected A549 cells were implanted into nude mice, there was no difference in tumor formation among all the groups. (2) After 5-FC administration, the tumor transfected with tissue-specific gene displayed a higher sensitivity to the drug than those treated with non-specific in vitro gene-therapy. (3) The tumor-bearing nude mice were randomized in a blind manner based on comparable size to receive the supernatant of recombinant retrovirus G1CEACDNa followed by 5-FC, and significant growth suppression could be observed. (4) Comparing to the group with injection of 5-fluorouracil (5-FU) alone, tissue-specific suicide gene therapy showed lower suppression to bone marrow.
CONCLUSIONSThe results suggest that tissue-specific suicide gene therapy may play an important role in individual treatment of lung cancer.
9.Detection and genetic analysis of TT virus in hemodialysis patients
Wu NI ; Hao REN ; Xiaohui MIAO ; Zhongtian QI ; Chaoyang YE
Academic Journal of Second Military Medical University 2001;22(4):331-333
Objective: To study the prevalence and pathogenesis of TT virus (TTV) in hemodialysis patients. Methods: Serum TTV DNA was tested in 69 hemodialysis patients from our hospital by nested-PCR using primers from a conservative region of TTV genenome, genetic analysis and detection of hepatitis C virus antibody (anti-HCV) and the levels of alanine aminotransferase (ALT) were also carried out simultaneously. Results: The overall prevalence of TTV viremia was 27.5%. The PCR-amplified gene fragment from one patient was sequenced, and its gene sequence homologies with GH1,TA278, TTVCHN1 and TTVCHN2 ranged from 89% to 100%, its deduced amino acid sequence ranged from 87% to 100%. There was no significant difference of TTV prevalence between anti-HCV positive and negative patients. No significant elevation of ALT was found in all patients. Conclusion: High prevalence of TTV infection is found among hemodialysis patients, and TTV infection has no significant association with HCV infection or elevation of ALT.
10.Screening antigenic epitopes from HCV core protein random peptide libraries displayed on phage
Wei PAN ; Zhongtian QI ; Xiaolan WU ; Xin PAN ; Qiuli CHEN ; Ping DU ; Xiang HE
Journal of Cellular and Molecular Immunology 2001;17(1):20-23
Aim To screen out the antigenic sequences from HCV core protein random peptide libraries displayed on phage and to explore a new way to screen the viral antigens. Methods The anti-HCV core antibody-positive serum was used to screen antigenic peptides from the HCV core protein random peptide libraries displayed on phage for 4 rounds. Detection of numbers of positive clones, positive rate of insertion of HCV random DNA and positive rate of hybridization with HCV core probes were used to evaluate the screening effects. The DNA sequences of 7 selected clones with positive hybridization were determined and analysed. Results Six out of 7 sequences are HCV core protein sequences, in which 5 were perfectly displayed,and one was possibly displayed. These sequences included several major HCV core antigenic epitopes. The remaining one was E.coli nrfa gene. Conclusion The phage display technique can be applied to study the viral antigenic peptides with the advantages of simple, accuracy and rapidity.

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