30 June 2023, Volume 27 Issue 3
    

  • Select all
    |
  • LANG Qiaoli, HUANG Nan, LI Liping, YANG Xi, LIU Chunlin
    Life Science Research. 2023, 27(3): 189-195.
    Abstract ( 505 ) Download PDF ( 1562 )   Knowledge map   Save
    To construct Huh-7 cell line stably expressing human angiotensin converting enzyme 2 (hACE2), the pWPXL-neo-hACE2 lentiviral vector without a fluorescent protein gene was constructed and co-trans-fected into HEK293T cells with psPAX2 and pMD2.G-VSVG to generate lentivirus. The obtained lentivirus particles were used to infect Huh-7 cells, and the cells that could stably express hACE2 protein were se-lected with hygromycin B. The expression of hACE2 in Huh-7 cells was detected by indirect immunofluo-rescence assay and Western-blot. Furthermore, the binding activity of Huh-7-hACE2 cells to the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) pro-tein was analyzed by flow cytometry, and the susceptibility of Huh-7-hACE2 cells to SARS-CoV-2 was further determined with SARS-CoV-2 pseudovirus. The identified Huh-7-hACE2 cells were finally used in evaluating the neutralizing capacity of SARS-CoV-2 neutralizing antibody REGN10987 against pseudovirus. The results showed that the hACE2 protein was successfully expressed in Huh-7 cells, and the expressed hACE2 protein could bind to SARS-CoV-2 S protein RBD. Compared with Huh-7 cells, Huh-7-hACE2 cells were more susceptible to SARS-CoV-2 pseudovirus infection, and the neutralizing activity evaluated with pseudovirus was similar to that with SARS-CoV-2 reported in the literature. In conclusion, the study demonstrated that the Huh-7 cell line stably expressing hACE2 was successfully established, and could be used to evaluate the activity of corona virus disease 2019 (COVID-19) therapeutic monoclonal antibody. The established cell line is a useful tool to study the pathogenesis of SARS-CoV-2 and develop antiviral drugs and vaccines.
  • ZHAO Shanshan, DING Suping, ZHU Xiuzhi, YUAN Fenghua, ZHOU Jiangying, JIE Yaliang, YUAN Zhidong
    Life Science Research. 2023, 27(3): 196-209.
    Abstract ( 445 ) Download PDF ( 1598 )   Knowledge map   Save
    Novel coronavirus (SARS-CoV-2) infection increases the risk of myocardial injury, and the mole-cular mechanisms of pathogenesis remain to be elucidated. Herein, the changes of RNA editing in SARS-CoV-2 infected cardiomyocytes (infection group) and uninfected cardiomyocytes (mock group) were compa-red to explore the effects of the viral infection on cardiomyocytes from the perspective of RNA editing. RNA-seq data (GSE150392) of SARS-CoV-2 infection of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) was downloaded from the GEO database, and RNA editing sites (RESs) were iden-tified using SPRINT software. The changes in editing levels of cardiomyocytes in the infection and mock groups were compared, editing sites were annotated, and relevant functional analyses were performed. A total of about 92 899 base substitutions were detected, of which 87 670 were identified as A-to-I RESs, and 78 978 of these A-to-I editing sites were found to appear in the Alu regions. A-to-I editing sites tend to be dis-tributed in clusters, mainly in regions such as introns and intergenic regions. By screening the differential RESs between the infection and mock groups in A-to-I editing, it was found that 102 of the significantly differential RESs are up-regulated and 94 are down-regulated in editing level. These genes with significant differences in RES are involved in the biological processes of viral infection related GO (Gene Ontology), which are mainly enriched in viral process, virus life cycle, and defense response to virus, et al., and their Kyoto Encyclopedia of Genes and Genomes (KEGG) functional pathways are also mainly related to viral infec-tion. In contrast, the 11 screened genes with high-quality A-to-I RESs and significant differences in editing levels between groups are enriched in endocytosis, cytokine-cytokine receptor interaction, proteosome, nico-tinate and nicotinamide metabolism, and ferroptosis, et al.. The results showed that SARS-CoV-2 infection affects A-to-I RNA editing in cardiac myocytes, and the occurrence of such RNA editing events is a re-sponse of host cardiac myocytes to the viral infection.
  • CAI Yuhao, ZHANG Tingting, CHENG Yangjian
    Life Science Research. 2023, 27(3): 210-216.
    Abstract ( 444 ) Download PDF ( 1550 )   Knowledge map   Save
    Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes corona virus disea-se 2019 (COVID-19), has been widely spread around the world for three years, and has spawned many va-riants. At present, the Omicron variant, which causes large-scale transmission in the world, has the charac-teristics of high infectivity and low toxicity, and has evolved into BA.4, BA.5, XBB and other subvariants. These strains can escape the vaccine-induced immunity, but neutralizing capacity elicited by vaccination can be restored to a certain degree by a booster dose. New vaccines and therapeutic drugs against the Omi-cron variant are available, providing better immunity and treatment methods for humans. This article sum-marized and analyzed the characteristics of Omicron strains, human immune mechanism against the infec-tion, current vaccine and drug development, and COVID-19 sequelae. It also made suggestions on how to cope with SARS-CoV-2 in the future.
  • XIAO Pei, LI Meiling, ZHOU Jianlin, ZHOU Chang
    Life Science Research. 2023, 27(3): 217-222.
    Abstract ( 463 ) Download PDF ( 1268 )   Knowledge map   Save
    Triple negative breast cancer (TNBC) is a highly malignant tumor with a poor prognosis, and the mutation frequency of p53 gene in TNBC patients is more than 80%. Gamabufotalin (CS-6), a skin secretion from Japanese common toad (Bufo japonicus), has been shown to have anticancer effects. However, the role and underlying mechanism of CS-6 against TNBC have not been reported. Herein, the effects of different concentrations of CS-6 on the proliferation of TNBC cell lines MDA-MB-468 and MDA-MB-231 were de-tected by MTT assay and clone formation assay. The effects of CS-6 on the expression of p53 and heat shock protein 90 (HSP90) at exogenous and endogenous levels were detected by Western-blot. The synergistic effect of CS-6 and the HSP90 inhibitor tanespimycin (17-AAG) on the proliferation of MDA-MB-468 and MDA-MB-231 cells was investigated by combination index and equivalent linearization analysis. The results showed that CS-6 significantly inhibited the proliferation of MDA-MB-468 and MDA-MB-231 cells. The mechanism of inhibition of the tumor cell proliferation may be related to degradation of HSP90/mutant p53 protein. In addition, CS-6 and HSP90 inhibitor 17-AAG had a synergistic inhibitory effect on the prolifera-tion of MDA-MB-468 and MDA-MB-231 cells.
  • LIU Yan, DU Yarong, SUN Kun
    Life Science Research. 2023, 27(3): 223-228.
    Abstract ( 413 ) Download PDF ( 1447 )   Knowledge map   Save
    Compared with the traditional monolayer culture, three-dimensional cell culture (TDCC) can more accurately simulate the growth environment of cells in vivo. In recent years, TDCC has been increasingly used in basic biological research. Epigenetic modification is closely linked to many cellular processes such as cell canceration. Radiation is one of the main means of tumor radiotherapy, the process of which is often accompanied by dynamic epigenetic changes, leading to DNA methylation, chromatin remodeling, histone modification and regulation of non-coding RNA. Exploring the mechanism of radiation-induced epigenetic regulation based on TDCC model is of great significance for treatment and protection of cancer patients.
  • WANG Dandan, WANG Jianfeng
    Life Science Research. 2023, 27(3): 229-236.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    To explore the diversity of bacterial community in Cobolidia fuscipes (Meigen), the V3~V4 regions of bacterial 16S rDNA in this insect species at different development stages were sequenced using Illumina MiSeq high-throughput sequencing platform. A total of 548 058 reads and 870 operational taxonomic units (OTUs) were obtained and annotated into 24 phyla, 51 classes, 92 orders, 175 families and 346 genera. The Chao, Ace and Shannon indices were 485.00~885.03, 485.09~877.55 and 2.90~4.31, respectively. The domi-nant bacterial phyla were Proteobacteria and Actinobacteria, dominant orders were Rhizobiales and Actino-mycetales, and dominant genera were unclassified Xanthobacteraceae, Pseudomonas and Leucobacter. However, the dominant genus in each sample was different. Pseudomonas occupied the highest proportion in eggs, unclas-sified Xanthobacteraceae had the highest relative abundance in the 1st to 3rd instar larvae and adults, Cory-nebacterium had the highest relative abundance in the 4th instar larvae, and Providencia had the highest rela-tive abundance in pupae. Principle component analysis (PCA) demonstrated that the composition of bacterial community in C. fuscipes (Meigen) was different at different development stages. This study provides a theoreti-cal basis for exploitation of microbial resources in C. fuscipes (Meigen) and also for biocontrol of the pest species.
  • JIANG Neng, WU Liyan, WU Qingjie, LIU Zhixiao
    Life Science Research. 2023, 27(3): 237-244.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Cocoons are a key life-history stage of leeches, so it is of great ecobiological significance to clarify their morphological features and chemical composition. Wuling cave leeches (Sinospelaeobdella wulingensis) are the type species of the newly-described genus Sinospelaeobdella, which breed on the roof wall of karstic caves, and differ from water and land leeches ecobiologically. In view of the particularity of Wuling cave leeches and the key role of cocoons in the adaptive evolution of leeches, morphology and chemical composi-tion of the cocoons were preliminarily studied. The results showed that: 1) Cocoons of Wuling cave leeches were roughly ellipsoidal, with a long diameter of (10.80±1.71) mm and a short one of (8.32±1.60) mm, and a weight of (135.3±76.8) mg; 2) From outside to inside, each cocoon had the outer layer (honeycomb) and in-ner layer of cocoon-wall, protein fluid and fertilized egg; 3) There were 17 kinds of hydrolyzed amino acids and 11 kinds of free amino acids in the cocoon, among which the relative contents of glutamic acid and ala-nine were the highest; 4) There were 68 kinds of trusted proteins identified in the cocoon, of which 27 were unidentified, and the remaining 41 could be divided into 6 categories, namely 5 antioxidant proteins, 12 en-zyme proteins, 6 cytoskeletal proteins, 12 binding proteins, 2 heat shock proteins and 4 other proteins. The morphological characteristics and chemical composition of the cocoons reflect long-term adaptation of leeches to cave ecological environments. The crucial role of the cocoon in the species evolution and population sur-vival of the leeches deserves further systematic investigation.
  • ZHANG Rongrong, KANG Xiaohu, XIE Fang, TANG Deping
    Life Science Research. 2023, 27(3): 245-252.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Quorum sensing (QS) is an important way of interaction between microbial populations and between microbial communities and plants. The signaling molecules that regulate the quorum sensing process are called autoinducers (AIs), which play important regulatory roles in bioluminescence, virulence factor secretion, spore formation, secondary metabolite synthesis, and biofilm formation. Autoinducing peptides (AIPs) are one kind of the quorum sensing signaling molecules that specifically communicate between bacterial communi-ties. This article reviewed the research progress in the source, structure, function and mechanisms of AIPs, and discussed their application potentials in plant growth promotion and biocontrol. It would be beneficial for better understanding the ecological function of microbial community and the molecular mechanism of in-teraction with plants, and may provide some help for the application of quorum sensing signaling molecules in ecological agriculture.
  • LIU Xia, WANG Siyi, RUI Chen, ZHU Lilin, LU Henglei, QIAO Junwen, SHENG Hua, SUN Xuye, JIANG Aili, REN Jin, XIAO Ying
    Life Science Research. 2023, 27(3): 253-266.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Environmental stimuli play a key role in the survival and evolution of wild animals. Cell receptors that assist in perception, including touch and temperature, reflect biodiversity. Merkel cells (MCs) are impor-tant sensory cells in touch-sensitive areas of vertebrates. Herein, literature research and data statistics were used to investigate the distribution of MCs with the help of PubMed, ScienceDirect and China National Knowledge Infrastructure (CNKI). The results showed that MCs had been observed in 77 species within 6 ani-mal classes including Mammalia, Amphibia, Aves, Reptilia, Cyclostomata and Actinopteri. The histopatholo-gical studies of MCs mainly focused on Mammalia, which accounted for 50% and mainly included human, monkey, rat, mouse, dog, cat, pig and rabbit, then followed by Amphibia and Actinopteri. In tissues and organs, MCs were found to be distributed in nose/vibrissae in 33 species, which accounted for 40%, suggesting that the nose and vibrissae are important parts for animals, especially wild animals, to perceive external mecha-nical stimuli. MCs were also studied in palate, perioral area, fingers/palms of hands/feet, lip, tongue, dorsal skin, gingiva, limbs, abdomen skin, eyes, cheeks, gills and other touch-sensitive sites of many species. Sys-tematically summarizing the distribution of MCs found in known species and studying the distribution of MCs in different species would be helpful to understand the role of environmental diversity in biodiversity.
  • ZENG Bingjie, JIANG Qing, ZHANG Yuping, XU Ye, LIU Rushi, WANG Qinglin, LIANG Xianghui
    Life Science Research. 2023, 27(3): 267-274.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Liver cancer is a common malignant tumor in clinic. Studies have shown that sterol O-acyltrans-ferase 1 (SOAT1), a key enzyme in cholesterol metabolism, is closely related to the occurrence and develop-ment of liver cancer. Therefore, SOAT1 has great potential as a biomarker for the cancer. Herein, the gene encoding SOAT1 was obtained from the Huh-7 liver cancer cell line. After prokaryotic expression and pro-tein purification, the purified recombinant SOAT1 protein was used as an antigen to immunize mice. A total of 5 anti-SOAT1 monoclonal antibody cell lines, named 1F3, 1G3, 1D6, 2F8 and 4D11, respectively, were obtained by using hybridoma and other technologies. Among them, 4D11 was able to recognize the SOAT1 protein in Huh-7 cells and clinical liver cancer tissue samples. Subsequently, 4D11 was sent to a biological company for sequencing, and the amino acid sequences of the antibody variable regions were obtained. In conclusion, an anti-SOAT1 monoclonal antibody was successfully prepared by recombinant SOAT1 protein, which would not only provide the raw materials for the subsequent application of SOAT1 protein in develo-ping liver cancer companion diagnostic kits, but also lay a foundation for the establishment of SOAT1-based liver cancer diagnostic methods.
  • JIN Jin, LIANG Xujun, BI Wu, ZHANG Pengfei
    Life Science Research. 2023, 27(3): 275-282.
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    A drug target is a specific structure in a cell, tissue or organ that interacts with a drug to make it work. Comprehensive identification of drug targets is essential to understanding the mechanism of action of drugs and their potential side effects. Currently, widely used drug target identification and validation tech-niques, such as activity-based protein profiling (ABPP), compound-centric chemical proteomics (CCCP), re-quire the modification of small-molecule compounds, which may reduce or alter the activity of drug mole-cules. Therefore, advanced techniques that do not require chemical modification of drug molecules, such as drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and thermal proteome profiling (TPP), are becoming important tools for drug target research. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a powerful tool for identification of drug target proteins. This paper reviewed drug target research techniques based on LC-MS/MS and future development of drug target identification.